ROLE OF BIOTECHNOLOGY IN TEXTILE AND PAPER & PULP INDUSTRIES BIOTECHNOLOGY IN PULP AND PAPER INDUSTRY The pulp

and paper industry is a large and growing portion of the worlds economy. Pulp and paper production has increased gl obally, as has the rate of paper consumption. In general, the industry is very capital-intensive with small prot margins. T his tends to limit experimentation, development, and incorporation of new technologies into mills. However, the pulp and paper industry is facing increasing pressure from environmental regulations. To keep up with the increasing demand for pulp and paper and to meet increasingly stringent environmental regulations, the industry has been constantly looking towards technological improvements. Over the past 20 years, research efforts in laboratories around the world have sought to apply biotechnology in industrial wood processing. This brief overview summarises different biotechnological applications of microbes and their enzymes in the pulp and paper industry which have been commercialised or are under development. It also offers a perspective on future developments. BIOPULPING- Biopulping is dened as the treatment of lignocellulosic materials with lignin -degrading fungi prior to pulping. It is getting closer to commercialisation. In the 1970s, Eriksson and co-workers at the Swedish pulp and paper research institute (STFI), Stockholm, launched a fairly comprehensive investigation that demonstrated that fungal pre-treatment of lignocellulosic materials could result in energy savings and strength improvements for mechanical pulping. Mechanical pulping involves th e use ofmechanical force to separate wood bres. Mechanical processes have high yield (up to 95 per cent) and produce paper with high bulk, good opacity and excellent printability. However, they are energy intensive (electricity use) and produce paper with relatively low strength and high colour reversion rate (tendency to turn yellow with time). The pulps from several wood species have high pitch content and therefore require ameliorating steps. Although the STFI research had limited success (it encountered difculties in scale-up), it provided valuable insights. A more comprehensive evaluation of biomechanical pulping was launched in 1987 at the US Departent of Agriculture (USDA) Forest Service. The consortium has established the economic feasibility of biopulping at pilot scale in connection with mechanical pulping. The two-week, environmentally friendly process increases mill throughput by 30 per cent or reduces the electrical energy requirement by at least 30 per cent at unchanged throughput. It also improves paper strength. Investigations at laboratory scale have sorted through the more than 30 variables associated with biopulping, including species and strains or fungi, inoculums form and amount, species of wood, wood chip size, environmental factors, effect of added nutrients, need to sterilise the chips, etc. Of several hundred species and strains of white-rot fungi examined to date, Ceriporiopsis subvermispora was found to be the best for both hardwood and softwood species. PITCH PROBLEMS- Pitch is the mixture of hydrophobic resinous materials found in many wood species and constitutes some 2-8 per cent of to al wood weight, depending upon the species and the time of year. It causes a number of problems in pulp and paper manufacture, including deposits on tile and metal surfaces, plugging of drains, discoloration of the felt, tears and other defects in paper, downtime for cleaning, etc. Traditional methods of controlling pitch problems include natural seasoning of wood before pulping and/or adsorption and dispersion of the pitch particles with chemicals in the pulping and papermaking processes, accompanied by adding ne talc, di spersants and other kinds of chemicals. During the past ten years or so, two biotechnological methods have been developed independently and are now been used industrially. In the late 1980s, scientists in Japan discovered that the treatment of mechanical (groundwood) pulps with lipases, which catalyse the hydrolysis of triglycerides, reduces pitch problems signicantly. In the ea rly 1990s, Sandoz Chemicals Corporation in the United States (now Clariant Corporation) introduced a new product for control of pitch in pulpwood chips, called Cartapip. Cartapip is a fungal inoculum of the ascomycete Ophiostoma piliferum. A water slurry of the fungal spores is sprayed onto wood chips as they are piled prior to pulping. The fungus invades the wood cells, degrading the pitch. Pitch, inc1uding toxic resin acids, is also metabolised quite effectively by lignin-degrading fungi in biopulping, thus offering an additional benet. FIBRE IMPROVEMENTS OR MODIFICATIONS- The structure and chemical composition of pulp bre surfaces are of paramount importance for paper strength and other properties. Due to the higher yields obtained with mechanical pulps as compared to chemical pulps, they have attracted growing interest. Sometimes chemical pulps are added to mechanical pulps to impart strength or other properties. With improvement of mechanical pulp bre properties, the use of chemical pulps can be reduc ed or eliminated.Enzymes have been used to improve physical properties of bres and might have a commercial role in future. Cellulases can enhance pulp brillation and thereby improve paper strength. They can reduce bre coarseness and increase paper densi ty and smoothness. However, they reduce viscosity and must be used with care. Xylanase preparations have also been reported to improve pulp brillation and bre bonding. With recycled bres, there is growing concern about the rate of water drainage on the paper machine. The speed of paper machine operation depends in part on the drainage rate ofwater out of the pulp mat. Drainage rates tend to be lower for recycled bres than for virgin bres so that there is a decr ease in the paper machine production rate as recycled bre content increases. It has been discovered that cellulases and hemicellulases can improve the drainage rates of recycled bres. Pilot and mill-scale testing has led to the commercial use of these enzymes as drainage aids. In the future, other enzyme-based processes could lead to cleaner and more efcient pulp and paper processing. Starch -modifying enzymes are sometimes used to improve paper quality. Enzymatic modication of starches is a cleaner process than chemical (oxidative) modication, as less energy is used and less waste is produced. Enzymetically modied starches at the wet end (size press) are applied in about 10 per cent of pa per production. DEINKING- Traditional deinking processes use NaOH, NaSO3, silicates and hydrogen peroxide for deinking oil-based printing materials such as newspapers and magazines. However, with the growing use of coating and new types of inks containing synthetic polymers in laser and xerographic printing, conventional deinking methods are inadequate for producing high-quality pulps. Recycling mills are therefore increasingly dependent upon mechanical devices to break down the larger non-ink particles to allow for removal by oatation or washing. Enzymatic techniques that allow for deinking of all kinds of recycled papers have recently been developed and commercialised.Cellulase acts on cellulose of wood fibres and facilitates the loosening of ink from fibre thus reducing the need of chemicals . BLEACHING OF KRAFT PULPS- The kraft process accounts for most of the worlds pulp production. Kraft pulping degrades and removes most of the lignin, without severely damaging the cellulose. Kraft pulps have a characteristic brown colour, which must be removed by bleaching before the manufacture of printing and writing or other products in which appearance is important. Kraft bleach plants use a variety of chemicals and treatment sequences to convert brown kraft pulp to white pulp. Traditionally, chlorination has been used, but because of consumer resistance and environmental regulations on chlorine bleaching, pulpmakers are turning to other bleaching chemicals (chlorine dioxide, oxygen, ozone, and peroxide), to extended pulping times (thereby lowering the pulp lignin content and decreasing bleaching chemical

requirements), and to other process modications. However, disadvantages associated with some of these methods are higher cost and/or greater danger of loss of pulp yield and strength as compared with chlorination. The essence of the process is a new enzyme better suited to the temperatures and pH found in pulp processing. The cost of the process is said to be the same as the conventional chlorine-intensive method. This is an example of the continuous improvement that characterises many biotechnological processes. Studies conducted in Finland show that hemicellulases (mainly xylanases) enhance pulp bleaching. These enzymes are now being used commercially in Scandinavia, Canada, the United States, and Chile. The treatment of kraft pulps with xylanases leads to sig nicant reduction in chemical consumption with almost no loss in pulp yield or quality. Biobleaching of acid bisulphite pulp with xylanases has also shown promise, with chemical savings of up to 51 per cent. Research is now being directed towards the discovery or engineering of enzymes that are more robust with respect to pH and temperature. Ligninases such as manganese-dependent peroxidase and laccases have also shown potential in pulp bleaching, but have not been used commercially. Both of these enzymes can achieve more substantial delignifying action than xylanase, but there are obstacles to be overcome before either enzyme can be used cost-effectively. There is currently no large-scale commercial source for either enzyme, so costs remain to be established. Current efforts have been to produce these enzyme cheaply enough so that the technology can become economically attractive. Also, genetic engineering of a fungus to produce a desired mixture of enzymes and their cosubstrate in situ may become more cost-effective than producing and applying the enzymes in separate steps. REDUCTION OF ORGANOCHLORINE COMPOUNDS IN BLEACH PLANT EFFLUENTS- Organochlorines have been a matter of concern in the pulp and paper industry for the last two decades. These compounds are produced mainly by the reactions between residual lignin present in wood bres and the chlorine and chlorine derivatives used for bleaching. Some of these compounds are toxic, mutagenic, and persistent; bioaccumulation causes harm to biological systems. Earlier measures taken by the pulp and paper industry to solve the chlorine problem focused on improving efuent treatment methods. Many physico -chemical methods have been used to treat bleach plant efuents, including precipitation with lime, alum and metal ions, and synthetic polymeric coagulants; adsorption on activated carbon, natural clays and polymeric adsorbents; membrane techniques; rapid ltration in soil; UV irradiation; and oxidation using oxygen, sulphur dioxi de, hydrogen peroxide and sodium hypochlorite. The problems underlying the physico-chemical treatments are those associated with cost and reliability. Today, R&D in this area has shifted towards improving the pulping process to decrease production of undesirable by-products. Biotechnological methods have the potential to eliminate or reduce the problems associated with physico-chemical methods. Biological treatments with bacteria or fungi are known to be effective in reducing the biological oxygen demand (BOD), the chemical oxygen demand (COD), and the toxicity of kraft pulp mills. Some enzymes also seem to have the potential to remove colour and adsorbable organic halogens from pulp and paper mill efuents. Peroxidase, laccase, etc., are the most important of these. Many factors have to b e considered in choosing an effective and commercial bleaching/treatment process that meets all the environmental guidelines. These processes are not used commercially. The most widely practised of the earlier biotechnologies are waste treatment processes. These are based in large part on the degradative activities of mixtures of aerobic and anaerobic micro-organisms, primarily bacteria. At present, cleaner production is largely achieved by process-integrated water treatment using biologically treated process water from the same production plant. Some 10-20 per cent of European paper producers reuse treated water in this way, so that there is zero discharge of wastewater. In the United States and Japan, a much smaller number of paper manufacturers use treated wastewater. REFINING OF THERMO MECHANICAL PULP (TMP) Refining is a mechanical process in which wood chips are separated to free fibres. The process used large amount of electricity . The enzyme cellulase acts on cellolose in wood fibres and softens wood chips to minimize refining time and electricity required. BIOFILM PROBLEMS- Biolms (slimes) in pulp and paper mills are a serious problem. They clog wires, pipes, and drains and conta minate the product itself, sometimes to the point of discoloration. They are controlled primarily through the use of biocides, some of which can be toxic to humans and other life forms. A signicant amount of research has gone into nding environmentally b enign control methods. Because the biolms are comprised of bacteria and fungi embedded in a matrix of extracellular polysaccharides, enzymes that hydrolyse the polymers have been studied. There is at least one commercial enzyme product, ED -l, a levulanase used by paper mills in the United States, Scandinavia, the United Kingdom, and Japan. Another promising method of controlling biolms is the introduction of non -lm-forming microbes that outcompete the biolm formers for substrates. It is likely that a combination of enzymes, friendly microbes, and dispersants will ultimately be used to lower or eliminate the use of biocides in pulp and paper mills.

Fig process in pulp and paper industry Fig 2. Table showing main characteristics of enzymes applied in various processes

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APPLICATION OF BIOTECHNOLOGY IN TEXTILE INDUSTRY

INTRODUCTION Retting of flax was the first biotechnological application in textile processing. More than 2000 years ago, micro-organisms grown on flax were used to achieve partial decortication in the extraction of linen fibres from flax stems. Amylases were the only enzymes applied in textile processing until the 1980s. These enzymes are still used to remove starch-based sizes from fabrics after weaving. Research on enzymatic applications in textile processing dates back to the beginning of the last century. During this period, the potential of proteolytic enzymes was assessed for the removal of wool-fibre-scales resulting in improved anti-felting behaviour. Despite the fact that investigations in this area are still on going, an industrial process has not yet been achieved. This is largely attributed to the heterogeneous nature of textile fibres and the unacceptable fibre strength losses incurred. With the advent of biological detergents in the 1960s, proteases made their way into detergent formulations specifically to remove organic protein-based stains (e.g. from egg, blood) from textile garments. Later in the 1970s, cellulases were found to add detergency during fabric washing and to remove fibrillation in multiple washes. Today, cellulases are included in many washing powders. Cellulases have also been employed to enzymatically remove fibrils and fuzz fibres and have also successfully been introduced to the cotton textile industry and later for lyocell processes. Further applications have been found for these enzymes to produce the aged look of denim and other garments. In May 2000, the First International Symposium on Biotechnology in the Textile Industry was held in Portugal and was attended by more than 150 participants from all over the world. The presentations given at this forum by scientists and delegates from industry reflected the enormous potential of Biotechnology in the textile fiel . Advances in biotechnology have made it possible to tailor special enzyme mixtures for specific applications. For example, amylases have been developed for desizing processes running at 100 C while cellulase monocomponents were identified to be superior to the native enzymes in several textile applications. Besides hydrolytic enzymes such as cellulases, amylases, pectinases (bioscouring) and proteases (wool finishing), other enzyme activities including oxidoreductases have been realised as powerful tools in various textile-processing steps. Several studies dealing with cellulases are presented in this special issue. By focusing on process development and the control of enzymatic fibre hydrolysis this research strives to find a balance between the beneficial effects of enzyme treatments and the potential strength losses. A number of investigations have dedicated their efforts to the phenomenon known as back staining, experienced during bio-stoning and bio-finishing of both cotton and linen fabrics. Other authors took the great challenge to elucidate the degradation mechanism of the natural substrates of cellulases. The effect of endoglucanases and cellobiohydrolases from different sources were used for these investigations together with components of these enzymes.The results of these contributions combined with information about the molecular architecture and specificities on soluble substrate of cellulases from different families will improve our understanding of the functioning of this interesting class of enzymes. Although fibres from cotton were the main target substrates for enzymatic modifications introduced in the last few years, enzymes also seem to have a potential for the improvement of fibres/fabrics from other sources such as flax and wool. Two interesting contributions show how enzymes can be used both in flax processing and analysis. Besides enzymes, the biopolymer chitosan can be used to improve the properties of wool. Biotechnological processes for the treatment of textile effluents can be grouped into two areas, microbial systems and enzymes. In microbial effluent treatment, a combination of anaerobic and aerobic steps seems to be beneficial in achieving sufficient detoxification. New more efficient treatment processes and their integration into textile finishing are discussed in several of the following papers. Ligninolytic enzymes such as laccases, lignin peroxidases and manganese peroxidases have been shown to decolourise textile dyes involving either polymerisation or degradation of dyes. The mechanisms of decolorisation and detoxification have been described for several dyes, including azo compounds. However, although some azo dyes were degraded with concomitan conversion of the azo group into molecular nitrogen

(harmless), these enzymes did not attack some dyes at all. It appears that the redox potential rather than steric effects seem to determine the degradation velocity. Another interesting application of enzymes for textile effluent treatment involves the use of catalases for the conversion of hydrogen peroxide present in bleaching effluents to oxygen and water. Both this process and enzymatic treatment of dyeing effluents have been shown to enable recycling of the process waters, especially when immobilised enzymes had been used. Industrial Processes in the Textile Industry-The textile industry is comprised of a diverse, fragmented group of establishments that produce andor process textile-related products (fiber, yarn, fabric) for further processing into apparel, home furnishings, and industrial goods. Textile establishments receive and prepare fibers; transform fibers into yam, thread, or webbing; convert the yarn into fabric or related products; and dye and fmish these materials at various stages ofproduction. The process of converting raw fibers into fineshed the apparel and nonapparel textile products is complex; thus, most textile mills specialize. Little overlap occurs between hitting and weaving, or among production of manmade, cotton, and wool fabrics. The primary focus ofthis section is on weaving and knitting operations, with a brief mention of processes used to make carpets. In its broadest sense, the textile industry includes the production of yam, fabric, and finish goods. This section focuses on the following four production stages, with a brief discussion of the fabrication of non-apparel goods: 1.yarn formation 2. fabric formation 3. wet processing 4. .fabrication Yarn Formation Textile fibers are converted into yam by grouping and twisting operations used to bind them together. Although most textile fibers are processed using spinning operations, the processes leading to spinning vary depending on whether the fibers are natural or manmade. Figure shows the different steps used to form yarn. Note that some of these steps may be optional depending on the type of yarn and spinning equipment used. Natural fibers, known as staple when harvested, include animal and plant fibers, such as cotton and wool. These fibers must go through a series of preparation steps before they can be spun into yarn, including opening, blending, carding, combing, and drafting. Manmade fibers may be processed into filament yarn or staple-length fibers (similar in length to natural fibers) so that they can be spun. Filament yarn may be used directly or following further shaping and texturizing. The main steps used for processing natural and manmade fibers into yam are below.

Flowchart of yarn formation process

Opening/Blending. Opening of bales sometimes occurs in conjunction with the blending of fibers. Suppliers deliver natural fibers to the spinning mill in compressed bales. The fibers must be sorted based on grade, cleaned to remove particles of dirt, twigs, and leaves, and blended with fibers from different bales to improve the consistency of the fiber mix. Sorting and cleaning is performed in machines known as openers. The opener consists of a rotating cylinder equipped with spiked teeth or a set of toothed bars. These teeth pull the unbaled fibers apart, fluffing them while loosening impurities. Because the feed for the opener comes from multiple bales, the opener blends the fibers as it cleans and opens them. Carding. Tufts of fiber are conveyed by air stream to a carding machine, which transports the fibers over a belt equipped with wire needles. A series of rotating brushes rests on top of the belt. The different rotation speeds of the belt and the brushes cause the fibers to tease out and align into thin, parallel sheets. Many shorter fibers, which would weaken the yarn, are separated out and removed. A further objective of carding is to better align the fibers to prepare them for spinning. The sheet of carded fibers is removed through a funnel into a loose ropelike strand called a sliver. Opening, blending, and carding are sometimes performed in integrated carders that accept raw fiber and output carded sliver.

. Combing. Combing is similar to carding except that the brushes and needles are finer and more closely spaced. Several card slivers are fed to the combing machine and removed as a finer, cleaner, and more aligned comb sliver. In the wool system, combed sliver is used to make worsted yam, whereas carded sliver is used for woolen yam. In the cotton system, the term combed cotton applies to the yam made from combed sliver. Worsted wool and combed cotton yarns are finer (smaller) than yam that has not been combed because of the higher degree of fiber alignment and fiuther removal of short fibers. Drawing. Several slivers are combined into a continuous, ropelike strand and fed to a machine known as a drawing frame (Wingate, 1979). The drawing frame contains several sets of rollers that rotate at successively faster speeds. As the slivers pass through, they are further drawn out and lengthened, to the point where they may be five to six times as long as they were originally. During drawing, slivers from different types of fibers (e.g., cotton and polyester) may be combined to form blends. Once a sliver has been drawn, it is termed a roving. Drafring. Drafting is a process that uses a frame to stretch the yam further. This process imparts a slight twist as it removes the yam and winds it onto a rotating spindle. The yarn, now termed a roving in ring spinning operations, is made up ofa loose assemblage of fibers drawn into a single strand and is about eight times the length and one-eighth the diameter of the sliver, or approximately as wide as a pencil (Wingate, 1979). Following drafting, the rovings may be blended with other fibers before being processed into woven, knitted, or nonwoven textiles. Spinning. 'The fibers are now spun together into either spun yams orfilament yams. Filament yams are made from continuous the strands of manmade fiber (e.g. not staple length fibers). Spun yarns are composed of overlapping staple length fibers that are bound together by twist. Methods used to produce spun yams, rather than filament yams, are discussed in this section. The rovings produced in the drafting step are mounted onto the spinning frame, where they are set for spinning. The yarn is first fed through another set of drawing or delivery rollers, which lengthen and stretch it still further. It is then fed onto a high-speed spindle by a yarn guide that travels up and down the spindle. The difference in speed of travel between the guide and the spindle determines the amountof twist imparted to the yarn. .The yarn is collected on a bobbin. Manmade fibers include 1) cellulosic fibers, such as rayon and acetate, which are created by reacting chemicals with wood pulp; and 2) synthetic fibers, such as polyester and nylon, which are synthesized from organic chemicals. Since manmade fibers are synthesized From organic chemicals, yam formation of manmade fibers does not involve the extensive cleaning and combing procedures associated with natural fibers. Manmade fibers, both synthetic and cellulosic, are manufactured using spinning processes that simulate or resemble the manufacture of silk. Spinning, in terms of manmade fiber production, is the process of forming fibers by forcing a liquid through a small opening beyond which the extruded liquid solidifies to form a continuous filament. Following spinning, the manmade fibers are drawn, or stretched, to align the polymer molecules and strengthen the filament. Manmade filaments may then be texturized or otherwise treated to simulate physical characteristics of spun natural fibers. Texturizing is often used to curl or crimp straight rod-like filament fibers to simulate the appearance, structure, and feel of natural fibers. Fabric Formation The major methods for fabric manufacture are weaving and knitting. Figure shows fabric formation processes for flat fabrics, such as sheets and apparel. Weaving, or interlacing yarns, is the most common process used to create fabrics. Weaving mills classified as broad woven mills consume the largest portion of textile fiber and produce the raw textile material from which most textile products are made. Narrow wovens, nonwovens, and rope are also produced primarily for use in industrial applications. Narrow wovens include fabrics less than 12 inches in width, and nonwovens include fabrics bonded by mechanical, chemical, or other means. Knitting is the second most frequently used method of fabric construction. The popularity of knitting has increased in use due to the increased versatility of techniques, the adaptability of manmade fibers, and the growth in consumer demand for wrinkle-resistant, stretchable, snug-fitting fabrics.

Weaving Weaving is performed on modern looms, which contain similar parts and perform similar operations to simple hand-operated looms. Fabrics are formed from weaving by interlacing one set of yarns with another set oriented crosswise. Satin, plain, and twill weaves are the most commonly used weave patterns. In the weaving operation, the length-wise yarns that form the basic structure of the fabric are called the warp and the crosswise yarns are called the filling, also referred to as the weft. While the filling yarns undergo little strain in the weaving process, warp yarns undergo much strain during weaving and must be processed to prepare them to withstand the strain Before weaving, warp yarns are frst wound on large spools, or cones, which are placed on a rack called a creel. The warp yarns are then unwound and passed through a sue solution (sizingklashing) before being wound onto a warp beam in a process known as beaming. The size solution forms a coating that protects the yarn against snagging or abrasion during weaving.

Knitting Knitted fabrics may be constructed by using hooked needles to interlock one or more sets of yams through a set of loops. The loops may be either loosely or closely constructed, depending on the purpose of the fabric. Knitted fabrics can be used for hosiery, underwear, sweaters, slacks, suits, coats, rugs, and other home furnishings. Knitting is performed using either weft or warp processes. In weft (or filling) hitting, one yam is carried back and forth and under needles to form a fabric. Yams run horizontally in the fabric, and connections between loops are horizontal. In warp knitting, a warp beam is set into the knitting machine. Yarns are interlocked to form the fabric, and the yarns run vertically while the connections are on the diagonal. Several'different types of machinery are used in both weft and warp knitting.

Weft knitting. Weft knitting uses one continuous yam to form courses, or rows of loops, across a fabric. There are three fundamental stitches in weft knitting: plain-knit, purl, and rib. On a machine, the individual yam is fed to one or more needles at a time. Weft knitting machines can produce both flat and circular fabric. Circular machines produce mainly yardage but may also produce sweater bodies, pantyhose, and socks. Flatbed machines knit full garments and operate at much slower speeds. The simplest, most common filing knit fabric is single jersey. Double knits are made on machines with two sets of needles. All hosiery is produced as a filling knit process. Warp Knitting. Warp knitting represents the fastest method of producing fabric from yarns. Warp knitting differs from weft knitting in that each needle loops its own thread. The needles produce parallel rows of loops simultaneously that are interlocked in a zigzag pattern. Fabric is produced in sheet or flat form using one or more sets ofwarp yams. Wet processing- Woven and knit fabrics cannot be processed into apparel and other finished goods until the fabrics have passed through several water-intensive wet processing stages. Wet processing enhances the appearance, durability, and serviceability of fabrics by converting undyed and unfinished goods, known as gray or greige (pronounced gri[zh]) goods, into finished consumers goods. Also collectively known as fmishing, wet processing has been broken down into four stages in this section for simplification: fabric preparation, dyeing, printing, and finishing. These stages, shown in Figure involve treating gray goods with chemical baths and often require additional washing, rinsing, and drying steps. Note that some of these steps may be optional depending on the style of fabric being manufactured. Typical wet processing steps for fabric

Fabric Preparation- Most fabric that is dyed, printed, or finished must be prepared, with the exception of denim and certain knit styles. Preparation, also known as pretreatment, consists of a series of various treatment and rinsing steps critical to obtaining good results in subsequent textile finishing processes. . In preparation, the mill removes natural impurities or processing chemicals that interfere with dyeing,

printing, and finishing. Typical preparation treatments include desizing, scouring, and bleaching. Preparation steps can also include processes, such as singeing and mercerizing, designed to chemically or physically alter the fabric. For instance, the mercerizing stage chemically treats the fabric to increase fiber strength and dye affinity, or ability to pick up dyes. This, in turn, increases the longevity of fabric finishes applied during finshing. Many ofthe pollutants from preparation result from the removal of previously applied processing chemicals and agricultural residues. , These chemical residues can be passed on to subsequent stages with improper preparation. Singeing. If a fabric is to have a smooth finish , singeing is essential. Singeing is a dry process used on woven goods that removes fibers protruding from yarns or fabrics. These are burned off by passing the fibers over a flame or heated copperplates. Singeing improves the surface appearance of woven goods and reduces pilling. It is especially useful for fabrics that are to be printed or where a smooth finish is desired. Pollutant outputs associated with singeing include relatively small amounts of exhaust gases from the burners. Desizing. Desizing is an important preparation step used to remove size materials applied prior to weaving. Manmade fibers are generally sized with water-soluble sizes that are easily removed by a hot-water wash or in the scouring process. Natural fibers such as cotton are most often sized with water-insoluble starches or mixtures of starch and other materials. Enzymes are used to break these starches into watersoluble sugars, which are then removed by washing before the cloth is scoured. Removing starches before scouring is necessary because they can react and cause color changes when exposed to sodium hydroxide in scouring. Scouring. Scouring is a cleaning process that removes impurities from fibers, yarns, or cloth through washing. Alkaline solutions are typically used for scouring; however, in some cases solvent solutions may also be used. Scouring uses alkali, typically sodium hydroxide, to break down natural oils and surfactants and to emulsify and suspend remaining impurities in the scouring bath. The specific scouring procedures, chemicals, temperature, and time vary with the type of fiber, yarn, and cloth construction. Impurities may include lubricants, dirt and other natural materials, water-soluble sizes, antistatic agents, and residual tints used for yarn identification. Typically, scouring wastes contribute a large portion of biological oxygen demand (BOD) loads from preparation processes. Bleaching. Bleaching is a chemical process that eliminates unwanted colored matter from fibers, yams, or cloth. Bleaching decolorizes colored impurities that are not removed by scouring and prepares the cloth for further finshing processes such as dyeing or printing. Several different types of chemicals are used as bleaching agents, and selection depends on the type of fiber present in the yam, cloth, or finished product and the subsequent finishing that the product will receive. The most common bleaching agents include hydrogen peroxide, sodium hypochlorite, sodium chlorite, and sulfur dioxide gas. Hydrogen peroxide is by far the most commonly used bleaching agent for cotton and cotton blends, accounting for over 90 percent of the bleach used in textile operations, and is typically used with caustic solutions. The bleaching process involves several steps: 1)The cloth is saturated with the bleaching agent, activator, stabilizer, and other necessary chemicals; 2) the temperature is raised to the recommended level for that particular fiber or blend and held for the amount of time needed to complete the bleaching action; and 3) the cloth is thoroughly washed and dried. Mercerizing. Mercerization is a continuous chemical process used for cotton and cotton polyester goods to increase dyeability, luster, and appearance. This process, which is carried out at room temperature, causes the flat, twisted ribbon-like cotton fiber to swell into a round shape and to contract in length. This causes the fiber to become more lustrous than the original fiber, increase in strength by as much as 20 percent, and increase its affinity for dyes. Dyeing Dyeing operations are used at various stages of production to add color and intricacy to textiles and increase product value. Most dyeing is performed either by the finshing division of vertically integrated textile companies, or by specialty dyehouses. Specialty dyehouses operate either on a commission basis or purchase greige goods and finish them before selling them to apparel and other product manufacturers. Textiles are dyed using a wide range of dyestuffs, techniques, and equipment. Dyes used by the textile industry are largely synthetic; typically derived from coal tar and petroleum-based intermediates. Dyes are sold as powders, granules, pastes, and liquid dispersions, with concentrations of active ingredients ranging typically from 20 to 80 percent. Printing Fabrics are often printed with color and patterns using a variety of techniques and machine types. Of the numerous printing techniques, the most common is rotary screen. However, other methods, such as direct, discharge, resist, flat screen (semicontinuous), and roller printing are often used commercially. Pigments are used for about 75 to 85 percent ofall printing operations, do not require washing steps, and generate little waste (Snowden-Swan, 1995). Compared to dyes, pigments are typically insoluble and have no affinity for the fibers. Resin binders are typically used to attach pigments to substrates. Solvents are used as vehicles for transporting the pigment and resin mixture to the substrate. The solvents then evaporate leaving a hard opaque coating. Finishing Finishing encompasses chemical or mechanical treatments performed on fiber, yam, or fabric to improve appearance, texture, or performance. Mechanical finishes can involve brushing, ironing or other physical treatments used to increase the luster and feel of textiles. Application of chemical finishes to textiles can impart a variety of properties ranging from decreasing static cling to increasing flame resistance. The most common chemical finishes are those that ease fabric care, such as the permanent-press, soil-release, and stain resistant finishes. Chemical finishes are usually followed by drying, curing, and cooling steps. Application of chemical finish are often done in conjunction with mechanical finishing steps .Selected mechanical and chemical finishing techniques are described below. Mechanical Treatments Hearsetting. Heatsetting is a dry process used to stabilize and impart textural properties to synthetic fabrics and fabrics containing high concentrations of synthetics. When manmade fibers are heatset, the cloth maintains its shape and size in subsequent finishing operations and is - stabilized in the form in which it is held during heat setting (e.g., smooth, creased, uneven). Textural properties may include interesting and durable surface effects such as pleating, creasing, puckering, and embossing. Heatsetting can also give cloth resistance to wrinkling during wear and ease-of-care properties attributed to improvements in resiliency and in elasticity. Pollution outputs may include volatile components of spin finishes if heatsetting is performed before scouring and bleaching processes. These components are introduced to the fabrics during the manufacture of synthetic fibers, when proprietary spin finish are applied to provide lubrication and impart special properties, such as antistatic, to the fiber.

Brushing and napping. Brushing and napping decrease the luster of fabrics by roughening or raising the fiber surface and change the feel or texture of the fabric (ATMI, 19971.). These processes involve the use of wires or brushes that pull individual fibers. Softening. Calendering, or ironing, can be used to reduce surface friction between individual fibers, thereby softening the fabric structure and increasing its sheen. In calendering, the fabric passes through two or more rolls. Typically, one roll is made of chilled steel, while the other is made of a softer material like cotton fihers. The steel roll may also be heated using gas or steam. Once goods pass through the machine they are wound up at the back of the machine. Opticalfinishing. Luster can be added to yarns by flattening or smoothing the surfaces under pressure. This can be achieved by beating the fabric surface or passing the fabric between calendering rolls. The luster can be further increased if the rolls are scribed with closely spaced lines. Shearing. Shearing is a process that removes surface fibers bypassing the fabric over a cutting blade. Compacting. Compacting, which includes the Sanforizing process, compresses the fabric structure to reduce stresses in the fabric. The Sanforizing process reduces residual shrinkage of fabrics after repeated laundering (Wingate, 1979). The fabric and backing blanket are fed between a roller and a curved braking shoe, with the blanket under tension. The tension on the blanket is released after the fabric and blanket pass the braking shoe. Compacting reduces the potential for excessive shrinkage during laundering. Chemical Treatments

Opticalfinishes. Optical fmishes added to either brighten or deluster the textile. Absorbent and soil release finishes. These finishes that alter surface tension and other properties to increase water absorbency or improve soil release. Softeners and abrasion-resistant finishes. ofteners and abrasion-resistant finishes are added to improve feel or to increase the ability of the textile to resist abrasion and tearing.

Application of biotechnology in textile industry

Fibre Preparation in textiles Linen is a cellulosic fibre obtained from the flax plant. These fibres are formed in the cortex between the lignified core and the outer layers of the stem, they are separated from the stems by retting, in which matrix components, mainly pectin and lignin are removed and the fibres are separated. Recently, considerable efforts have been put to use enzymes in the retting process to control the process to produce linen fibres of consistent quality. Pre-treatment of the flax with sulphur dioxide gas brings about sufficient breakdown of the woody straw material to speed up enzyme retting whilst preventing excessive bacterial or fungal deterioration of the fibre. The carbonization process in which vegetable matter in wool is degraded by treatment with strong acid and then subjected to mechanical crushing can, in principle, be replaced by selective enzyme degradation of the impurities. Fabric Preparation Desizing using amylase enzymes has been well established for many years. However, there is still considerable scope for improving the speed, economics and consistency of the process, including the development of more temperature stable enzymes as well as a better understanding of how to characterize their activity and performance with respect to different fabrics, sizes, and processing conditions, e.g. for pad batch as opposed to jigger desizing. The current application in the textile industry involves mainly hydrolases and now to some extent is Oxidoreductase. The Tables 1 and 2 exemplify such textile applications. Table 1: Application of Hydrolase Enzyme in Fabric Preparation S.No Enzyme Substrate Textile Application Name 1 Amylase Starch Starch desizing 2 Cellulase Cellulose 1. Stone wash-Bio-polishing (Bio-singeing) 2. Bio finishing for handle modification 3. Carbonization of wool 3 Pectinase Pectin Bio scour replacing caustic 4 Catalase Peroxides In situ peroxide decomposition without any rinse in bleach bath 5 Lipases Fats and oils Improve hydrophilicity of PET in place of alkaline hydrolysis

Table 2: Application of Oxidoreductase in Fabric Preparation S.No Enzyme Name Substrate 1 Laccase Colour Chormophore and pigments

Peroxidases

Colour Chromophore & pigment

Textile Application 1.Discoloration of coloured Chromophore effluent 2. Bio-bleaching of lignin containing and pigments fibres like kenaf and jute 3. Bio-bleaching of indigo in denim for various effects Bio-bleaching of wood pulp

An already established application is the use of catalase enzymes to breakdown residual hydrogen peroxide after, for example, pre bleach of cotton that is to be dyed a pale or medium shade. Reactive dyes are especially sensitive to peroxide and currently require extended rinsing and/or use of chemical scavengers. The enzyme catalase is added after oxidative bleaching and allowed to react for 15 minutes at 30 C- 40 C. It degrades the residual peroxide in water and oxygen. The results obtained were compared with the conventional process and it was found that the outcome of the enzymatic process was excellent. The best suitable conditions are the temperature range of 20 C- 60 C, pH 5-10 and the application time is 10 min to 15 min. Finishing Bio-stoning and the closely related process of bio-polishing are perhaps attracting most current attention in the area of enzyme processing. They are also an excellent illustration of how different industry structural and market considerations can affect the uptake of enzyme technology. Conventional stone washing uses abrasive pumice stones in a tumbling machine to abrade and remove particles of indigo dyestuff from the surfaces of denim yarns and fabric. Cellulase enzymes can also cut through cotton fibres and achieve much the same effect without the damaging abrasion of the stones on both garment and machine. Disadvantages can include degradation of the fabric and loss of strength as well as 'back staining'. A slight reddening of the original indigo shade can also occur. Now processors are learning to play more sophisticated tunes such as achieving a peach skin finish by use of a combination of stones and natural cellulase. Bio-polishing employs basically the same cellulose action to remove fine surface fuzz and fibrils from cotton and viscose fabrics. The polishing action thus achieved helps to eliminate pilling and provides better print definition, colour brightness, surface texture, drapeability, and softness without any loss of absorbency. Bio-polishing can be used to clean up the fabric surface after the primary fibrillation of a peach skin treatment and prior to a secondary fibrillation process which imparts interesting fabric aesthetics. A weight loss in the base fabric of some 3-5% is typical but reduction in fabric strength can be controlled to within 2-7% by terminating the treatment after about 30-40 min using a high temperature or low pH 'enzyme stop'. One area that still poses problems is that of tubular cotton finishing. Here, the fibre residues tend to be trapped inside the fabric rather than washed away. Wool Processing Applications The international wool secretariat (IWS) together with, Novo, been developing the use of protease enzymes for a range of wool finishing treatments aimed at increased comfort (reduced prickle, greater softness) as well as improved surface appearance and pilling performance. The basic mechanisms are found closely parallel to those of bio-polishing. The improved enzyme treatments will allow more selective removal of parts of the wool cuticle, there by modifying the luster, handle and felting characteristics without degradation or weakening of the wool fibre as a whole and without the need for environmentally damaging prechlorination treatment. Other Protease Applications Protease enzymes similar to those being developed for wool processing are already being used for the degumming of silk and for producing sand washed effects on silk garments. Treatment of Silk-Cellulosic blend is claimed to produce some unique effects. Proteases are also being used to wash down printing screens after use in order to remove the proteinaceous gums, which are used for thickening of printing pastes. Textile After-care Enzymes have been widely used in domestic laundering detergents since the 1960s. Some of the major classes of enzymes and their effectiveness against common stains are summarized in Table 3 Table 3: Types Of Enzymes and Their Effectiveness Against Various Stains Enzymes Effective For Proteases Grass, Blood, Egg, Sweat stains Lipases Lipstick, Butter, Salad oil, Sauces Amylases Spaghetti, Custard, Chocolate celluloses Colour brightening, Softening, Soil removal Early problems of allergic reactions to some of these enzymes have now largely been overcome by the use of advanced granulation technology. Modern enzyme systems have reduced the use of sodium perborate in detergents by 25% along with the release of harmful salts into the environment. However, enzymes still have to make a corresponding impact upon the commercial laundering market. One of the problems here has been the level of investment in 'continuous-batch' or tunnel washers. These typically afford a residence time of 6-12 min which is not long enough for present enzyme systems to perform adequately. More efficient methods of 'enzyme kill' are also required because of the extent of water recycling in modern washers. Role In Waste Treatment Natural and enhanced microbial process has been used to treat waste materials and effluent streams from the textile industry. Conventional activated sludge and other systems are generally well able to meet BOD and related discharge limits on most cases. The industry faces some specific problems like colour removal from dyestuff effluent and handling of toxic wastes including PCPs and heavy metals. The synthetic dyes are designed in such a way that they become resistant to microbial degradation under the aerobic conditions. Also, the water solubility and the high molecular weight inhibit the permeation through biological cell membranes. Anaerobic processes convert the organic contaminants principally into methane and carbon dioxide and usually occupy less space, treat wastes containing up to 30 000 mg/l of COD, have lower running costs and produce less sludge. Natural fibre sources Several possibilities exist for producing entirely new fibre materials, so called biopolymers, using biotechnological process routes, naturally occurring polyester etc. PHB is produced by bacterial fermentation of a sugar feed stock and commercially available as 'Biopol'. The polymer is stable under normal conditions but biodegrades completely in any microbial active environment. Other biopolymers with textile potential include polylactates and polycaprolactones, which are investigated for medical applications. Bacterial Cellulose

The specialty papers and nonwovens are produced based on bacterially grown cellulose fibres. These are extremely fine and resilient and are used as specialized filters, odour absorbers and reinforcing blends with aramids. Genetically Modified Micro-Organisms Attempts have been made to transfer certain advantageous textile properties into microorganisms where they can be more readily reproduced by bulk fermentation processes. The spider DNA is transferred into bacteria with the air of manufacturing proteins with the strength and resilience of spider silk for use in bulletproof vests. Dyestuffs And Intermediates Attempts have been made to synthesize bacterial forms of indigo as well as fungal pigments for use in the textile industry. Certain micro fungi are capable of yielding up to 30% of their biomass as pigment. Potential non-textile applications include food industry colourants. Biotechnology For Tissue Engineering and Medical Textiles The application of polymer materials in medicine for producing various implants such as vascular prosthesis, heart valves, sutures etc, is one of the most significant achievements in contemporary surgery. Controlled revascularisation of the epidermal tissue is the key issues in tissue regeneration involving the use of porous and naturally occurring polymeric scaffolds on which cells are seeded. Textile structures gain importance as scaffolds to grow biological tissues in in-vitro. Thus tissue engineering also forms the integral aspect of biotechnology. Advantages of using enzymes in textiles 1. Weight loss during processing of fabric is minimised by enzymes. 2. Cotton processing by enzyme catalysis is possible. 3. Production cost is reduced.(water and energy) 4. Starting point for future research and development. 1. 2. Limitations of using enzymes in textiles Although time required for processing is more the lost can be compensated by time required between processes. First trail of cotton processing are in semi continuous way. INTRODUCTION: Bioprocess in leather industry. The meat processing industry generates hides of dead animals which would have caused environmental problems in disposal, had it not been for the leather industry. Fortunately the leather industry makes use of these hides to process it further and make leather. Although the leather industry takes care of this environmental problem and generates employment, the processing of hide to leather itself generates a fair amount of pollutants. That is because, the conventional processing of leather involves the use of chemicals and the maximum amount of solid wastes like lime and chrome sludge and noxious gases( like hydrogen sulphide )are generated during the leather making processes. It is in these areas that biotechnology through the use of enzymes has played a key role in refining the process of leather making. Today several of the chemicals used in leather processing have been substituted with enzymes. This has made the entire process of tanning hides to become more efficient and quicker. So today proteases, lipases and amylases are used in leather manufacturing. Advantages When enzymes are used in leather processing it conveys certain advantages such as: Water usage is high in conventional leather processing which is about 30 to 40 liters per kg of hide processed. The use of enzymes reduces this requirement considerably. The effluent discharges (both gaseous and aqueous) in leather processing using the conventional route (without using enzyme s) contributes to dissolved solids (chromium, lime, sulphides and sulphates etc) and Biological Oxygen Demand (BOD), and Chemical Oxygen Demand (COD). However, using biotech processes helps in reducing COD by 80%, chromium by 85% and Total Dissolved Solids by 85%. Different stages of leather processing There are three distinct stages of leather processing, namely, preparation for tanning, tanning, and finishing. Each of these stages involves several other steps. Some of these steps like soaking, liming, bating and degreasing involving a biotech perspective will be discussed in detail in this article.

Curing the hide

This is the first step which entails treating the flayed hide with brine. If this is not done, then there is the chance of the flayed hide getting putrefied. Conventionally, the hide was soaked in brine to remove unwanted parts of the hide and the skin. Now enzymes can do the job better as it can provide better soaking effect, because they can re-hydrate the hides better and quicken up the entire process. One example of such an enzyme is Specialty Enzyme's SEBsoak product. Biocides have also been found useful in curing the hides but this is not environment friendly. Despite all this, salt curing is still the predominant way of curing and biotech hasn't made inroads for curing the hide process.

Soaking
In this stage, the hides are washed and soaked in surfactants and other compounds mostly anti-microbial. The intention is to help in the further processing to leather. The conventional process of soaking uses sodium tetrasulphide plus a surfactant, in which case the soaking process will take nine hours. But proteases and lipases used along with surfactants can reduce the time required for soaking to five hours. Examples of proprietory enzymes used in soaking process are: a) Palkosoak which is a mixture of protease and lipase suitable for alkaline conditions b) Palkosoak ACP which is again mixture of protease and lipase that suits acidic conditions.

Liming
After soaking, the next step is liming operation. It may be that soaking would not have made the skins swollen to the required degree, so liming is done precisely to achieve desired swelling of skin. Conventionally this is done with milk of lime, resulting in swelling of the collagen structure, so the fiber bundles can be opened up. The idea is to remove the keratinous matter and remove proteins like mucins and the ultimate quality of leather depends on this process. Although not many enzymes are used in the liming process as of now, there are some proprietory enzymes that can stabilize the hide by removing all proteinous matter of non-leather origin from the hide. An example is SEBbate Acid that makes the hide smoother for fabrication and dyeing by ensuring smoother grain and pliability of the hide.

De-hairing
This entails making the hides free of hairs and furs. The conventional method is to use sulphide to eliminate keratin but the problem is it produces effluents with high COD. Instead, proteolytic enzymes of bacterial and fungal origin can now be used that will do the job by attacking the protein matter at the hair base. This obviates the need for sodium sulfide, and the process does not produce toxic wastes. Moreover, enzymatic process is far quicker. De-hairing can be done using extra cellular protease secreted by Bacillus isolate, by enzymes secreted by Rhizopus oryzae or by using alkaline protease from Alcaligenes faecalis. An example of a proprietory enzyme used in de-hairing process is Palkodehair which is a protease enzyme that works in alkaline conditions. Another example is SEB lime, which is biodegradable and eco friendly. The swell regulating properties of this enzyme results in better grain smoothness of leather.

Bating
The idea of bating is to make the leather soft and supple (to bring out the grain and give flexibility) suitable for tanning, normally achieved by striking the leather with metal and wooden rods so that residues of proteins and epidermis are removed, or sometimes by using the manure of pigeon or hen. Bating increases the stretch of leather, removes swelling and produces silky grain. Now proteolytic bating enzymes of pancreatic or bacterial origin are used for bating under alkaline conditions. It works by diffusion of the enzymes into the hides. Trypsin and alkaline proteases are commonly used. An example of a proprietory enzyme suitable for bating is Palkobate that works best in alkaline conditions.

Degreasing
It is essential to remove the fatty substances that escape the liming and other processes. Else it will result in uneven dyeing and finishing as for example cause waxy patches in leathers. The conventional method is acqueous emulsification, solvent extraction and pressure degreasing. When enzymes of lipase type are used, they will rupture fat cell membranes and cause triglyceride splitting thereby facilitating degreasing process.

Tanning
This is the last stage in making leather which involves introducing a tanning agent in the hides. Enzymes are not directly involved in this stage.

Waste processing
Trypsin and proteolytic enzymes are used in further processing chrome tanned waste from tanneries. Conclusion As you can see from this article, there is a paradigm shift in leather processing from chemical driven processes to enzyme driven processes. The key to using enzymes in leather processing is that it shouldn't damage or dissolve the keratin in the hides, but it should have the ability to hydrolyze casein, elastin, albumin and other non-structured proteins which are not required in the hide for leather making. Current biotech research in leather manufacturing has generated technologies for non-lime enzyme assisted de-hairing for cow hides, enzymatic dehairing for goatskin and sheepskin and a unique bio-driven three step tanning technique.

Nutraceutical
Nutraceutical, a portmanteau of the words nutrition and pharmaceutical, is a food or food product that reportedly provide s health and medical benefits, including the prevention and treatment of disease. Health Canada defines the term as "a product isolated or purified from foods that is generally sold in medicinal forms not usually associated with food. Such products may range from isolated nutrients, dietary supplements and specific diets to genetically engineered foods, herbal products, and processed foods such as cereals, soups, and beverages. The term nutraceutical was originally defined by Dr. Stephen L. DeFelice, founder and chairman of the Foundation of Innovation Medicine (FIM), Crawford, New Jersey. Examples are beta-carotene and lycopene. Classification of nutraceuticals Nutraceuticals is a broad umbrella term used to describe any product derived from food sources that provides extra health benefits in addition to the basic nutritional value found in foods. There are multiple different types of products that may fall under the category of nutraceuticals. Dietary supplements-A dietary supplement is a product that contains nutrients derived from food products that are concentrated in liquid or capsule form. The Dietary Supplement Health and Education Act (DSHEA) of 1994 defined generally what constitutes a dietary supplement. A dietary supplement is a product taken by mouth that contains a "dietary ingredient" intended to supplement the diet. The " dietary ingredients" in these products may include: vitamins, minerals, herbs or other botanicals, amino acids, and substances such as enzymes, organ tissues, glandulars, and metabolites. Dietary supplements can also be extracts or concentrates, and may be found in many forms such as tablets, capsules, softgels, gelcaps, liquids, or powders. Functional foods - Functional food are designed to allow consumers to eat enriched foods close to their natural state, rather than by taking dietary supplements manufactured in liquid or capsule form. Functional foods have been either enriched or fortified, a process called nutrification. This practice restores the nutrient content in a food back to similar levels from before the food was processed. Sometimes, additional complementary nutrients are added, such as vitamin D to Milk. All functional foods must meet three established requirements: foods should be (1) present in their naturally-occurring form, rather than a capsule, tablet, or powder; (2) consumed in the diet as often as daily; and (3) should regulate a biological process in hopes of preventing or controlling disease Medical Food- Medical foods arent available as an over-the-counter product to consumers.[14] The FDA considers medical foods to be formulated to be consumed or administered internally under the supervision of a physician, and which is intended for the specific dietary management of a disease or condition for which distinctive nutritional requirements, on the basis of recognized scientific principles, are established by medical evaluation. Medical foods can be ingested through the mouth or through tube feeding. Medical foods are always designed to meet certain nutritional requirements for people diagnosed with specific illnesses. Medical foods are regulated by the FDA and will be prescribed/monitored by medical supervision. FarmaceuticalsAccording to a report written for the United States Congress entitled "Agriculture: A Glossary of Terms, Programs, and Laws", (Farmaceuticals) is a melding of the words farm and pharmaceutical s. It refers to medically valuable compounds produced from

modified agricultural crops or animals (usually through biotechnology). Proponents believe that using crops and possibly even animals as pharmaceutical factories could be much more cost effective than conventional methods (i.e., in enclosed manufacturing facilities) and also provide agricultural producers with higher earnings BIOPROCESSING OF NUTRACEUTICALS 1-Utilization of metabolites of plant origin for manipulation of gut flora in rumen in favour of CLA production as also on secretion of herbal nutraceutical directly in milk by adding suitable herbs/herbal extracts in the feed. 2-Value addition to whey, a by product of dairy industry, by maximizing production of GOS from whey based on isolating and using highly enhancedgalactosyltransferase activity of selected microbial strains for. MODE OF ACTION OF NUTRACEUTICALS: WHEY PROTIENS: Whey is the natural by-product of the cheese-making process (it is the liquid part of milk that remains after the manufacture of cheese). It is a complete protein, with all the essential amino acids and with the highest protein quality rating among all proteins. The biological components of whey demonstrate immune-enhancing, antioxidant, antihypertensive, anti-tumor, hypolipidemic, antiviral and antibacterial properties. Whey Protein Isolate (like the ISM whey protein) is the most pure and concentrated form, and delivers more essential amino acids to the body when compared to other proteins on a gram-to-gram basis. Dosage varies. Athletes, and those who use it as a protein supplement, may take up to 10-25 grams or more a day. Some recent experiments in rodents indicate that the antitumor activity of the dairy products is in the protein fraction and more specifically in the whey protein component of milk. We and others have demonstrated that whey protein diets result in increased glutathione (GSH) concentration in a number of tissues, and that some of the beneficial effects of whey protein intake are abrogated by inhibition of GSH synthesis. Whey protein is particularly rich in substrates for GSH synthesis. We suggest that whey protein may be exerting its effect on carcinogenesis by enhancing GSH concentration. The glutathione (GSH) antioxidant system is the principal protective mechanism of the cell and is a crucial factor in the development of the immune response by the immune cells. Experimental data demonstrate that a cysteine-rich whey protein concentrate represents an effective cysteine delivery system for GSH replenishment during the immune response. Animal experiments showed that the concentrates of whey protein also exhibit anticancer activity. They do this via the GSH pathway, the induction of p53 protein in transformed cells and inhibition of neoangiogenesis OMEGA 3AND 6 FATYACID:Fish oils: rich in EPA (Eicosapentaenoic Acid... Omega 3 Oil) Flax oil: rich in ALA (Alpha-Linolenic Acid.... Omega 3 Oil)... body converts into EPA Nuts: GLA (Gamma Linoleic Acid.... Omega 6 Oil) Oral administration of a supplement rich in omega-3 fatty acids for 5 d before surgery may improve not only preoperative nutritional status but also preoperative and postoperative inflammatory and immune responses in patients who have cancer. Anti-cachectic: EFAs prevent & reverse wasting syndrome. EFAs induce apoptosis (suicide) in cells. EFAs slow metastasis EFAs impair tumour angiogenesis CATEGORIES OF NUTRACEUTICALS- Nutraceuticals are non-specifi c biological therapies used to promote wellness, prevent malignant processes and control symptoms. They are categorized as follows: 1.Based on chemical constituents (a) Nutrients - Substances with established nutritional functions, such as vitamins, minerals, amino acids and fatty acids. Common nutrients and their associated health Benefits are Vitamin E used for treatment of Parkinsons disease. Vitamin D used for treatment of tuberculosis Vitamin B used for Alzheimer disease. (b) Herbals -Herbs or botanical products as concentrates and extracts. Common herbs are: Evening prime rose oil- for treatment of atopic eczema.Garlic- having antibacterial and antifungal activity used for the treatment of weight loss.Ginger- having positive iontropic activity.Ginkgo- used for the treatment of post thrombic syndrome. (c) Dietary Supplement- The dietary supplement health and education act of 1994 defines the Dietary supplements as the products administered through mouth that contain a dietary ingredient intended to add something to the foods you eat.

Examples of dietary supplements are black cohosh- for menopausal symptoms, ginkgo biloba- for memory loss, and glucosamine/chondroitin for arthritis. They also serves specific functions such as sports nutrition, weight-loss supplements and meal replacements. Supplement ingredients may contain vitamins, minerals, herbs or other botanicals, amino acids, enzymes, organ tissues, gland extracts, or other dietary substances. They are available in different dosage forms, including tablets, capsules, liquids, powders, extracts, and concentrates 2. Traditional and Non- Traditional neutraceutical Wide variety of neutraceutical foods are available in the market which falls in the category of traditional foods and non traditional foods. (a) Traditional Neutraceutical Under the category of traditional Neutraceutical comes food in which no change to the food are made; It is simply natural, whole foods with new information about their potential health qualities. There has been no change to the actual foods, other than the way the consumer perceives them.Many fruits, vegetables, grains, fish, dairy and meat products contain several natural components that deliver benefits beyond basic nutrition, such as lycopene in tomatoes, omega-3 fatty acids in salmon or saponins in soy. Even tea and chocolate have been noted in some studies to contain health-benefiting attributes. Tomatoes and salmon are two types of food that researchers have found to contain benefits beyond basic nutrition - in this case, lycopene and omega-3 fatty acids, respectively. (b) Nontraditional Neutraceutical They are the outcome from agricultural breeding or added (nutrients and/or ingredients such as Orange juice fortified with calcium, cereals with added vitamins or minerals and flour with added folic acid are nontraditional neutraceutical. Agricultural scientists successfully have come up with the techniques to boost the nutritional content of certain crops. Research currently is being conducted to improve the nutritional quality of many other crops. TYPES OF DISEASES NUTRACEUTICALS USED Cardiovascular diseases 1. Anti-oxidants, Dietary fibers, Omega-3 poly unsaturated fatty acids, Vitamins, minerals for prevention and treatment of CVD. 2. Polyphenol (in grape) prevent and control arterial diseases 3. Flavonoids (in onion, vegetables, grapes, red wine, apples, and cherries) block the ACE and strengthen the tiny capillaries that carry oxygen and essential nutrients to all cells. Diabetes 1. Ethyl esters of n-3 fatty acids may be beneficial in diabetic patients. 2. Docosahexaenoic acid modulates insulin resistance and is also vital for neurovisual development. 3. Lipoic acid, an antioxidant, for treatment of diabetic neuropathy. 4. Dietary fibers from psyllium have been used for glucose control in diabetic patients and to reduce lipid levels in hyperlipidemia. Obesity 1. 2. Cancer 1. 2.

Herbal stimulants, such as ephedrine, caffeine, ma huang-guarana, chitosan and green tea help in body weight loss. Conjugated linoleic acid (CLA), capsaicin, Momordica Charantia (MC) possesses potential anti obese properties. Flavonoids which block the enzymes that produce estrogen reduce of estrogen-induced cancers. To prevent prostate/breast cancer a broad range of phyto-pharmaceuticals with a claimed hormonal activity, called phyto-estrogens is recommended.

Allergy Quercet (found in Onions, red wine and green tea) reduce the inflammation that results from hay fever, bursitis, gout, arthritis, and asthma. Alzheimers disease -carotene, curcumin, lutein, lycopene, turmeric etc may exert positive effects on specific diseases by neutralizing the negative effects oxidative stress mitochondrial dysfunction, and various forms of neural degeneration. Parkinsons disease Vitamin E in food may be protective against Parkinsons disea se. Creatine modifies Parkinsons disease features as measured by a decline in the clinical signs. Bioprocessing of Functional Food. FUNCTIONAL FOOD. -Functional food is a food where a new ingredient(s) (or more of an existing ingredient) has been added to a food [1] and the new product has an additional function (often one related to health-promotion or disease prevention). The general category of functional foods includes processed food or foods fortified with health-promoting additives, like "vitamin-enriched" products. Products considered functional generally do not include products where fortification has been done to meet government regulations and the change is not recorded on the label as a significant addition ("invisible fortification"). An example of this type of fortification would be the historic addition of iodine to table salt, or Vitamin D to milk, done to resolve public health problems such asrickets. Fermented foods with live cultures are considered functional foods with probiotic benefits. Functional foods are part of the continuum of products that individuals may consume to increase their health and/or contribute to reducing their disease burden. "Functional Food is a Natural or processed food that contains known biologically-active compounds which when in defined quantitative and qualitative amounts provides a clinically proven and documented health benefit, and thus, an important source in the prevention, management and treatment of chronic diseases of the modern age". It was debated at the 9th International Conference on "Functional Foods and Chronic Diseases: Science and Practice" at the University of Nevada, Las Vegas on March 15-17, 2011.Functional Food Center has adopted a new definition of functional food . Functional foods are an emerging field in food science due to their increasing popularity with health-conscious consumers and the ability of marketers to create new interest in existing products.

CURRENT TRENDS 1. IODINE to TABLE SALT Recommended dietary allowance of iodine is 150mg/100ml.

Since 1983, Tata chemicals has been manufacturing Vacuum Evaporated Iodized Tata salt. 2. Vitamin D fortification- In US he milk is fortified with 100IU/cup. Other products such as yogurt, cheese, orange juice etc are also fortified with milk. Amul calci plus is fortified with natural calcium and pregnant women from 150mg/day to 400-600mg/day.

FUNCTIONAL BEVERAGES Energy drinks- An energy drink is a type of beverage which is purported to boost mental or physical energy. There are various brands and varieties of energy drinks. They generally contain large amounts of caffeine and other stimulants. Many also contain sugar or other sweeteners, herbal extracts and amino acids and may or may not be carbonated. In the UK, Lucozade Energy was originally introduced in 1929 as a hospital drink for "aiding the recovery;" in the early 1980s, it was promoted as an energy drink for "replenishing lost energy." One of the first energy drinks introduced in America was Dr. Enuf. Its origins date back to 1949, when a Chicago businessman named William Mark Swartz was urged by coworkers to formulate a soft drink fortified with vitamins as an alternative to sugar sodas full of empty calories. He developed an "energy booster" drink containing B vitamins, caffeine and cane sugar. After placing a notice in a trade magazine seeking a bottler, he formed a partnership with Charles Gordon of Tri-Cities Beverage to produce and distribute the soda. [1]Dr. Enuf is still being manufactured in Johnson City, TN and sold sparsely throughout the nation. Ingredients:Energy drinks generally contain methylxanthines (including caffeine), B vitamins, and herbs. Other commonly used ingredients arecarbonated water, guarana, yerba mate, aa, and taurine, plus various forms of ginseng, maltodextrin, inositol, carnitine, creatine,glucuronolactone, and ginkgo biloba. Some contain high levels of sugar, and many brands offer artificially sweetened 'diet' versions. A common ingredient in most energy drinks is caffeine (often in the form of guarana or yerba mate). Caffeine is the stimulant that is found in coffee and tea. Energy drinks contain about three times the amount of caffeine as cola. Twelve ounces of Coca-Cola Classic contains 35 mg of caffeine, whereas a Monster Energy Drink contains 120 mg of caffeine. Benefits of energy drinks:Caffine- Stimulant, significant improvements in mental and cognitive performances as well as increased subjective alertness. B-vitamins- they maintain the metabolism, immune system enhancements and cell growth. Herbs- They have medicinal properties for early recovery. HEALTH SPPLEMENTS Malt energy drinks- High protein drinks Malt is germinated cereal grains that have been dried in a process known as "malting". The grains are made to germinate by soaking in water, and are then halted from germinating further by drying with hot air. Malting grains develops the enzymes required to modify the grain's starches into sugars, including monosaccharides such as glucose or fructose, and disaccharides, such as sucrose ormaltose. It also develops other enzymes, such as proteases, which break down the proteins in the grain into forms that can be used by yeast. Bioprocess involved:Malting is the process of converting barley into malt, for use in brewing or distilling, and takes place in a maltings, sometimes called a malthouse, or a malting floor. The sprouted barley is kiln-dried by spreading it on a perforated wooden floor. Smoke, coming from an oasting fireplace (via smoke channels) is then used to heat the wooden floor and the sprouted grains. The temperature is usually around 55 C (131 F). A typical floor maltings is a long, single-story building with a floor that slopes slightly from one end of the building to the other. Floor maltings began to be phased out in the 1940s in favour of "pneumatic plants". Here, large industrial fans are used to blow air through the germinating grain beds and to pass hot air through the malt being kilned. Like floor maltings, these pneumatic plants are batch processes, but of considerably greater size, typically 100 ton batches compared with 20 ton batches for floor malting. The malting process starts with drying the grains to a moisture content below 14%, and then storing for around six weeks to overcome seed dormancy. When ready, the grain is immersed or "steeped" in water two or three times over two or three days to allow the grain to absorb moisture and to start to sprout. When the grain has a moisture content of around 46%, it is transferred to the malting or germination floor, where it is constantly turned over for around five days while it is air-dried. The grain at this point is called "green malt". The green malt is then kiln-dried to the desired colour and specification.[8] Malts range in colour from very pale through crystal and amber to chocolate or black malts. PROBIOTICS. Probiotic organisms are live microorganisms that are thought to be beneficial to the host organism. According to the currently adopted definition by FAO/WHO, probiotics are: "Live microorganisms which when administered in adequate amounts confer a health benefit on the host". Lactic acid bacteria (LAB) and bifidobacteria are the most common types of microbes used as probiotics; but certain yeasts and bacilli may also be used. Probiotics are commonly consumed as part of fermented foods with specially added active live cultures, such as in yogurt, soy yogurt, or as dietary supplements. Probiotics are also delivered in fecal transplants, in which stool from a healthy donor is delivered like a suppository to an infected patient.

Etymologically, the term appears to be a composite of the Latin preposition pro ("for") and the Greek adjective (biotic), the latter deriving from the noun (bios, "life"). At the start of the 20th century, probiotics were thought to beneficially affect the host by improving its intestinal microbial balance, thus inhibiting pathogens and toxin producing bacteria. Today, specific health effects are being investigated and documented including alleviation of chronic intestinal inflammatory diseases,] prevention and treatment of pathogen-induced diarrhea, urogenital infections,, and atopic diseases. To date, in those cases which the European Food Safety Authority has investigated health claims that are made about probiotic products, it states that the evidence provided is insufficient to establish a cause and effect relationship between the consumption of the products containing probiotics and the claimed health benefits. Example- YAKULT Yakult is a fermented milk drink containing over 6.5 billion probiotic (friendly) bacteria - Lactobacillus caseistrain Shirota. Unlike most bacteria found in normal yoghurt, these probiotic bacteria are able to survive the acidic environment of the stomach and reach the intestines alive In the intestine they change the environment to favour the growth of beneficial bacteria and suppress the growth of harmful bacteria

Bioprocess involved:

1.Mixing of Raw Ingredients Live Lactobacillus casei Shirota strain is cultured in a 'seed tank' in our laboratory. Skim milk powder is mixed with sugar, glucose and filtered, sterilised water to make a sweet milky solution. 2. Sterilisation The sweet, milky solution is sterilised at a high temperature for a short time, destroying any bacteria that may be present. The solution is then transferred to a 6,500-litre culture tank via a closed system of pipes and valves. 3. Culture Tank The temperature of the tank is reduced to 37C and live Lactobacillus casei Shirota strain is added. The solution is allowed to ferment for about one week until the number of Lactobacillus casei bacteria reaches an ideal concentration. 4. Mixing and Storage Tank The concentrate is transferred to a 12,000-litre mixing and storage tank. The tank is chilled to around 2 C. Sterilised flavours, syrup solution, vitamins and calcium are added to the concentrate. Prior to bottling, the concentrate is diluted with filtered, sterile water. 5. Injection Blow Moulding Machine The plastic bottles are produced on site. The bottles are made from polystyrene. 6. Bottling and Packaging The bottles are wrapped with individual bottle labels. Then, they are filled with Yakult, capped with a foil lid, sealed and transferred along the conveyor belt to the packaging facility. Single bottles of Yakult are sorted into groups of five and shrinkwrapped in polypropylene film. Ten "5-packs" are grouped together and wrapped again in polyethylene film and then heat shrunk, forming a 'carton' of 50 Yakult bottles. 7. Refrigeration Room Finished products are kept refrigerated before delivery to stores. 8. Yakult Quality Assurance Yakult maintains a comprehensive quality assurance program in order to ensure that our product is of the highest quality. For this reason, samples are collected for laboratory analysis throughout the production process to confirm that the quality assurance measures in place have been effective.

Our testing involves more than 150 samples per production run, upon which a total of more than 200 tests are conducted. These determineLactobacillus casei numbers, check for potential contaminants, microbiological quality, composition, acidity, physical attributes and taste. In addition, each bottle is inspected for undesirable markings and incorrect printing. Quality assurance measures are in place to maintain standards for personnel and factory hygiene, equipment cleaning, processing methods and parameters, and product handling. Yakult's quality assurance utilizes a system called "Hazard Analysis and Critical Control Points" (HACCP). The principles of HACCP are internationally recognised as an excellent method for assuring stringently high standards. 9. Waste Management Cleaning - Yakult adheres to a comprehensive hygiene and sanitation program, following a cleaning program that is predominantly governed by CIP (Cleaning in Place). Steam, an environmentally friendly cleaner, is used to sterilize the pipes and tanks. A single phase chemical cleaner is used, reducing the numbers of chemicals introduced into the drains. Chlorine based chemicals are not used. Solid wastes - The amount of solid waste is relatively small. Liquid wastes - Any liquid waste goes into a holding tank in our water treatment facility. The acidity (pH) of the water is adjusted with acid or alkaline before being released into the sewage system.

DISEASE RESISTANCE A plant disease is any abnormal condition that alters the appearance or function of a plant. It is a physiological process that affects some or all plant functions. Disease may also reduce yield and quality of harvested product. Disease is a process or a change that occurs over time. It does not occur instantly like injury. Clare Kenag (1974): Plant disease as infectious or non-infectious. Infectious plant disease: Abnormal plant conditions caused by biological agencies eg fungi, bacteria, viruses and nematodes and are capable of being transmitted from diseased plants to health plants to cause disease on the latter under favourable environmental conditions. Non-infectious plant disease: An abnormality on plant cause by agencies such as nutrient deficiencies, hail, wing, frost lightning and other external factors which are non-infectious and their effects are not transmissible from affected to health plants. The Disease Triangle

Different Pathogens Causing diseases

Plant resistance mechanism- Two broad mechanisms: Race-specific or gene-for-gene resistance Non-specific or broad spectrum resistance: 1. Systemic acquired resistance (SAR) Induced systemic resistance

Race specific or gene-for gene resistance Elicitation of race-specific resistance; genetic incompatibility H. Flor described the genetics that underlie race-specific or gene-for-gene resistance of flax rust with its host flax (Flor, 1956). Many pathogens exhibit gene-for-gene resistance on their hosts.

Plant responses to pathogen attack Few terms: 1. 2. R-genes- responsible for providing resistance to plants against a diverse range of pathogens viz. mycoplasma, bacteria, fungi, nematodes, protozoa, parasites etc. Avr genes- avirulence genes- these genes encodes any determinant of specificity of interaction with host eg. PAMPs or MAMPs (Pathogen or microbe associated molecular patterns) Gives the pathogen an avirulent phenotype or a host plant that carries the corresponding R gene. HR- hyper sensitive response- mode of manifestation of resistance to a pathogen- localized induced cell death in the host plant at the site of infection. Mechanism Plant recognition of specific molecules (elicitors) produced by pathogen (Coded directly or indirectly by avirulence genes )

3.

Elicitor molecules interacts with receptors (encoded by R genes)

Cascade of host genes get activated leading to HR and inhibition of pathogen growth.

Fig. Gene-for-gene interactions specify plant disease resistance. Resistance is only expressed when a plant that contains specific R gene that has the corresponding avirulence gene. All the other combinations lead to lack of recognition by the host and the result is disease. Gene for gene system involving HR are seen in all involving intracellular obligate pathogens (viruses and mycoplasma) as well as intercellular facultative and obligate pathogens (bacteria, fungi, nematodes). Physiological features of HR-Rapid oxidative burst -Ion fluxes characterized by K+ - H+ exchange. -Cellular decompartmentalization. -Cross-linking and strengthening of plant cell wall. -Production of antimicrobial compounds (Phytoalexins)

-Induction of pathogenesis related (PR) protein such as chitinases and glucanases. ->HR and other necrotic reactions trigger a subsequent response i.e. SAR (Subsequent acquired resistance) that acts non-specifically throughout plant. Example of gene-for-gene interaction in disease resistance Interaction between potato and its leaf mold pathogen cladosporum flavum. Two c.flavum avirulence genes avr9 and avr4 encode precursors for elicitor peptides that specifically elucidated HR in tomato plants that have corresponding r R genes cf-9 and cf-4 respectively. 2. NON-Specific or Broad Spectrum Resistancei) Systemic acquire Resistance (SAR): -Triggered by HR and other necrotic reactions. - Confers quantitative protection against broad spectrum of microorganisms in a manner comparable to innate immunity of mammals. - Resistance is expressed locally at the site of primary inoculation but also systematically in tissues remotely located from initial treatment. This form of induced resistance is called SAR by Ryals et al. in 1992. Mechanism:

-Plant uses pattern recognition receptors (PRRs) to recognize conserved microbial signatures (elicitors). -This interaction triggers an immune response. If interaction is compatible, disease occurs. But if interaction is incompatible, plant cell death occurs and accumulation of salicylic acid (SA) takes place. -SAR is associated with induction of wide range of genes (called PR or Pathogenesis related genes), which gets activated upon SA accumulation. -In response to SA, positive regulator protein NP1 moves to the nucleus where it interacts with TGA transcription factor to induce defense gene expression thus activating SAR (PR genes). PR genes- Pathogenesis related proteins (PRs) were first described in 1970 in tobacco leaves infected with TMV. PRs are plant proteins that accumulate after pathogen attack. Examples are-PR-1 -PR-1,-1,3-glucanases (PR2) -Chitinases (PR3) -PR-4 -Osmotin(PR-5) -In tobacco, overexpression of PR-1 significantly increase resistance against infection by Perenospora tabacina and Phytopthora var. nicotinae.

ii) Induce systemic resistance(ISR) -Another form of induced resistance. -Not associates with typical SAR genes, thus suggesting that SA accumulation not requires for ISR. -ISR is manifested to plant by other microorganisms like rhizobacter or other bacterial strains present in soil, in association with plant. -In case of Rhizobacteria, which promotes growth in plant; suppresses disease through antagonism between bacteria and soil borne pathogens as well as by inducing systemic resistance in plant against, both root and foliar pathogens. -The generally non-specific character of induced resistance constitutes an increase in the level of basal resistance to several pathogens simultaneously, which is of benefit under natural condition where multiple pathogens may be present. -Some strains show specificity, indicating specific recognition between bacteria and plants at the root surface. E.g. In pseudomonas strains inducing systemic resistance in Arabidopsis, the O-antigenic side chains of bacterial outer membrane Lipopolysaccharide, acts as inducing determinant. -SA is not necessary for ISR. Instead, rhizobacteria mediated ISR is dependent on Jasmonic acid (JA) and ethylene signaling in plant. -Experimentally, when Arabidopsis is inoculated with a pathogen; leaves expressing SAR exhibit a primed expression of SA while leaves expressing ISR are primed to express JA/ethylene, but not SA-responsive genes. ConclusionThus a combination of SAR and ISR can increase protection against pathogens that are resisted through both pathways as well as extend protection to broader spectrum of pathogen than ISR and SAR alone. CULTURAL METHODS Cultural controls are the oldest methods that have been used to manage pest populations. However, with the development of synthetic pesticides these controls were rapidly abandoned or de-emphasized and research on them was largely discontinued. Because cultural controls are preventative rather than curative they are dependent on long-range planning. Also, because they are dependent on detailed knowledge of the bio-ecology of the crop-pests-natural controls-environment relationships, most of which, in the past, were poorly understood, the results were very variable, and it was often difficult to evaluate their effectiveness. STRATEGIES ON WHICH CULTURAL PRACTICES ARE BASED: i) Make the crop or habitat unacceptable to pests by interfering with their oviposition preferences, host plant discrimination or location by both adults and immatures. ii) Make the crop unavailable to the pest in space and time by utilizing knowledge of the pest's life history, especially its dispersal and overwintering habits. iii) Reduce pest survival on the crop by enhancing its natural enemies, or by altering the crop's susceptibility to the pest. To design and implement cultural controls, it is necessary to have accurate knowledge of crop and pest biology, ecology and phenology, and of the weak links in pest-crop interactions. ADVANTAGES:-Cultural controls are generally the cheapest of all control measures because they usually only require modifications to normal production practices. Sometimes they do not even require extra labour, only careful planning. Often they are the only control measures that are profitable for high acreage of low value crops. Cultural controls are dependable, and are usually specific. Of major importance is the fact that they do not possess some of the detrimental side effects of pesticides, namely the creation of resistance to pesticides, undesirable residues in food, feed crops and the environment, and the killing of non-target organisms. DISADVANTAGES:- Cultural controls require long-term planning for greatest effectiveness and they need careful timing. They are often based on the substitution of knowledge and skills for purchased inputs and, as such, are more demanding on the farmer's competence. They may be effective for one pest but may be ineffective against a closely related species. Effectiveness of cultural controls is difficult to assess and they do not always provide complete economic control of pests. Some cultural controls have adverse effects on fish and wildlife and may also cause erosion problems. Chemical Plant Disease Control A plant disease is a physiological condition reflected by its structural abnormality that is harmful to the plant or to any of its parts and also reduces its economic value. Plant diseases may be temporary or permanent in natu re. The plant diseases may be caused by the micro-organisms; including fungi, bacteria, and viruses, etc. It may also be caused by the

physiological conditions like high or low temperatures, excess or lack of soil moisture and aeration, deficiency or excess of plant nutrients, soil acidity or alkalinity, etc. It has been found that wind, water, seed, diseased plants, cuttings, tubers, animals, insects and soil help to spread the diseases. Therefore, the control of plant diseases usually lies in preventing and curing the infectio n through various means. The use of chemical sprays, dusts or seed treatment for protecting plants from the ravages of the pathogens is not an innovation of the 20th century. The first landmark in the control of fungal disease of plants was the discovery by Anton de Bary that the causal agents of many plant disease are fungi. Even when a crop has been grown from pathogen free soil and the crop seems superficially healthy, the use of fungicides in some circumstances become unavoidable. Resistant varieties of potato are attacked by late blight if rain occurs during tuber formation. To control this disease use of fungicides become essential to avoid any loss of the crop. Therefore, even in the case of resistant varieties, judicious use of chemical protection will prolong the life of resistance in the variety. For killing different groups of pathogens, different types of chemicals are required. The chemicals mainly used for controlling diseases are: fungicides for killing fungus, bactericide for killing barteria, nematicides for killing nematodes, and viricides for killing viruses. The aim and of use of chemicals in plant disease control are to create a toxic barrier between the host surface or tissue and the pathogen and; to eradicate the pathogen present at a particular site on the host, such as seed, foliage, roots etc.

Classification of Chemicals
There are many chemicals which are available for plant disease control but all are not equally safe, effective and popular. The success of any chemical depends on the selection of suitable chemical and its use at appropriate time and place and its proper application. The chemicals can be broadly classified on the basis of their mode of action against pathogen and type of pathogen. A. According to mode of action Depending upon their mode of action the chemicals can be grouped as protectants, eradicants and therapeutants. They are briefly discussed as under. 1. Protectants Protectants are prophylactic in their behaviour. These may be applied to seeds, plant surfaces or the soil but cannot penetrate deep into plant tissues. Therefore, they act outside the plant parts as a cover to check the invasion by the pathogen. They include thiram, captan, agallol, zineb, sulphur, ceresan and streptocyclin. 2. Eradicants Eradicants help to eradicate the dormant or active pathogen from the host completely. They are effective in checking the entry (by covering surfaces) and penetrating in the tissue killing pathogen in the host plant. These chemicals can be used as protectant as well as eradicant. Examples are lime, sulphur, organo mercurials, such as phenyl mercury acetate, methoxy ethyl mercury chloride. 3. Therapeutants Therapeutants is an agent that inhibits the growth and development of a disease already entered in a plant, when applied appropriately. This therapy can be achieved by physical means, such as solar energy treatment or hot water treat- ment, but quite often by chemical means and is called chemo- therapy. Usually the chemo-therapeutants are systemic in their action, i.e. they enter the plants and affect deep-seated infection. e.g. plantvax, vitavax, brassicol, dithane M-45, dithane Z- 78, bordeaux mixture, streptocyclin and agromycin. B. According to type of pathogen Based on the type of pathogens, the chemicals can be grouped as (1) fungicides (2) bactericides and (3) nematicides 1. Fungicides The chemicals which kill fungus are called fungicides. Most of the plant diseases are caused by fungus only. Some diseases remain on the surface of the plant while others are deeply seated. These fungicides can again be classified as systemic and non-systemic based upon their action in plant. Systemic fungicides The chemicals which on, application, are absorbed and spread into the whole system of plant to kill fungus in all its stages are called systemic fungicides. They are absorbed by the roots and transported acropetally to the xylem. They are usually not transported downwards through the phloem tissue. These fungicides can kill pathogen present inside as well as outside the plant. Some of the common systemic chemicals are bavistin, carboxin, plantvax, and cycosin: b. Non- systemic fungicides Non-systemic or contact fungicides are the chemicals which do not enter inside the plant tissue, but kill the pathogen by surface contact. Some of the examples are captan, ziram, thiram, terbam, zineb, Maneb, bordeaux mixture, burgundy mixture etc. 2. Bactericides

The chemicals which kill bacteria are known as bactericides. The chemicals used for killing bacteria are steptocyclin, streptomycin, agrimycin-l00 and endomycin. These are generally known as antibiotics. 3. Nematicides The chemicals which kill the nematodes are called nematicides. The nematicides commonly used are, nemaphos, D.D, DBCP, carbofuran, phorate, telone, terracur, terracur-P, methon M-sodium and vapam. III. Anti-pathogen Chemicals The chemical substances that help to retard the activity of pathogens like fungus, bacteria and nematodes are said to be anti-pathogen chemicals. A. Functions The anti-pathogen chemicals are expected to perform three major functions. They include: i. reduction in inoculum density or eradication of inoculum from source of growth, multiplication and survival, ii. inactivaion or destruction of the pathogen when it lands on the treated surface, and iii. cure of the diseased plant. Majority of the modern chemicals are used as "protectants" to prevent infection of susceptible plant surfaces by a pathogen. This helps in eradication of the dormant inoculum present on the surface. However, very few chemicals are available to eradicate an established active plant disease infection.

B. Desired characters- A basic quality of fungicides, bactericides and nematicides is that they possess differential toxicity. They have
different effect on different groups of living organisms. Certain chemicals are more or less toxic to all or most pathogens while others are toxic to only a specific group of pathogens. The term "broad spectrum" and "narrow spectrum" of chemicals is used to indicate the range of pathogens affected by a particular chemical. In general, the anti-infection chemicals or fungicides having following characters are supposed to be ideal. i. High toxicity for the pathogen at low concentration. ii. Absence of toxicity for the host, man and animals. iii. Slow or no loss of toxicity in storage. iv. Retention of toxicity on dilution. v. Good spreading quality on host surface. vi. High tenacity on the host surface i.e. it should be retained on the surface. IV. Chemicals for Disease Control. The number of chemicals available for plant disease control are many although all are not equally safe, effective, and popular. Some of the important categories of chemicals used for disease control are given here A. Sulphur fungicides B. Copper fungicides C. Mercury fungicides D. Quinone fungicides E. Benzene fungicides F. Hetero-cyclic nitrogen compound Methods of Chemical Application Basically, the chemicals are applied by the methods of foliar or vegetative application, soil application and seed treatment. This depends on the type of vegetation, locality, infestation of diseases etc. These methods are described in detail here. A. Foliar application Application of chemicals on the stem, leaves flowers and fruits is called vegetative or foliar application. It is carried out in the form of spraying, dusting and pastes. 1. Spraying

It is more prevalent for fungal disease than for bacterial diseases of the foliage. Wettable chemicals available in solution or emulsified form are used for spraying to provide a protective covering over the surface before arrival of. the pathogen. As a protective layer such toxicants destroy the spores before or after their germination and infection of the host is checked. Many systemic fungicides are used as spray materials. The spraying is influenced by the volume i.e. a. High volume spray -more than 400 l/ha b. Low volume spray -5 -400 l/ha c. Ultra low volume -less than 5 l/ha spray 2. Dusting Insoluble or non-suspendable materials are used for dusting on the foliage. Fungicides and bactericides are more effective as spray rather than as dust. Dusting is advisable only in very wet weather which favours sticking of chemical of host surface. 3. Pasting and painting Chemicals are occasionally mixed with water, alcohol, or other carriers and applied as paint or paste on injured surface or parts of the plant. When trees are pruned application of paste or paint is necessary. B. Soil application The purpose of chemical soil treatment is to eradicate or reduce the inoculum density of soil-borne plant pathogens. However, complete eradication of pathogens from soil with chemical treatments is neither feasible nor desired. One cause of failure of chemicals to completely eradicate pathogens from soil is their rapid degradation by physical, chemical and biological conditions of soil. Most fungicidal soil treating , chemicals have the ability to diffuse through soil particles. In the control of nematodes volatile soil fumigants are used. The chemical soil treatment can be done in any of the following ways. 1. Drenching of soil with solution of suspension Chemicals are applied to soil surface in quantities sufficient to wet 10-15 cm depth of the soil. The treatment can be applied before or after planting of seeds. 2. Broadcasting of dusts, powders of granules Non-volatile fungicides are mixed with soil or fertilizers and broadcast on the soil surface. Light ploughing or harrowing is done to mix the chemical in sufficient depth. This method of soil treatment is too expensive. 3. Furrow application Furrow application is most common practice followed to apply dusts and granules. This method is possible in crops planted in furrows such as potato and sugarcane. 4. Fumigation Soil fumigation is usually done to control plant parasitic nematodes. The chemicals used for this purpose are usually volatile and on coming in contact with soil moisture release gases which diffuse in the soil and kill the larvae of the nematode. Application of these highly toxic volatile substance is recommended some days or weeks before actual planting of the crop. Special equipments are required for their application. The success of soil treatment depends on many factors. Proper ploughing of the land is necessary before-treatment. The soil should be well pulverised and should be neither too dry nor too wet. If organic manures are to be added that should be done after fumigation. Many soil fumigants are not effective in soils containing high organic matter content C. Seed treatment Seeds, tubers, bulbs and other propagating materials are given chemical treatment for eradication of pathogens present on them and for preventing their rot in the soil after planting. Sometimes, for seed borne disease seed treatment is the only method of control. Example covered smut of barley and grain smut of jowar. On the basis of their mode of action the seed treatment chemicals can be of three types. 1. Seed disinfestants 2. Seed protectants BIOLOGICAL METHODS Biological control is a method of controlling pests (including insects, mites, weeds and plant diseases) using other living organisms. It relies on predation, parasitism, herbivory, or other natural mechanisms, but typically also involves an active human management role. There are three basic types of biological pest control strategies: importation (sometimes called classical biological control), augmentation and conservation.

Natural enemies of insect pests, also known as biological control agents, include predators, parasitoids, and pathogens. Biological control agents of plant diseases are most often referred to as antagonists. Biological control agents of weeds include herbivores and plant pathogens. Types of biological pest control There are three basic types of biological pest control strategies: importation (sometimes called classical biological control), augmentation and conservation Importation Importation (or "classical biological control") involves the introduction of a pest's natural enemies to a new locale where they do not occur naturally. This is usually done by government authorities. In many instances the complex of natural enemies associated with a pest may be inadequate, a situation that can occur when a pest is accidentally introduced into a new geographic area, without its associated natural enemies. These introduced pests are referred to as exotic pests and comprise about 40% of the insect pests in the United States. There are many examples of successful importation programs, including: Joseph Needham noted a Chinese text dating from 304AD, Records of the Plants and Trees of the Southern Regions, by Hsi Han, which describes mandarin oranges protected by biological pest control techniques that are still in use today. One of the earliest successes in the west was in controlling Icerya purchasi, the cottony cushion scale, a pest that was devastating the California citrus industry in the late 19th century. A predatory insect Rodolia cardinalis (the Vedalia Beetle), and a parasitoid fly were introduced from Australia by Charles Valentine Riley. Within a few years the cottony cushion scale was completely controlled by these introduced natural enemies. Augmentation Augmentation involves the supplemental release of natural enemies, boosting the naturally occurring population. Relatively few natural enemies may be released at a critical time of the season (inoculative release) or millions may be released (inundative release). An example of inoculative release occurs in greenhouse production of several crops. Periodic releases of the parasitoid,Encarsia formosa, are used to control greenhouse whitefly, and the predatory mite Conservation The conservation of existing natural enemies in an environment is the third method of biological pest control. Natural enemies are already adapted to the habitat and to the target pest, and their conservation can be simple and cost-effective. Lacewings, lady beetles, hover fly larvae, and parasitized aphid mummies are almost always present in aphid colonies BiotechnologicalapproachindiseasemanagementBiotechnological approach A) Marker assisted plants breeding in disease management includes

Different markers and application in disease resistance Achievements

B) Tissue culture methods Somaclonal variation Somatic hybridization

C)Genetic engineering (Transgenics) D) Meristem Tip culture (for virus free planting material) Marker assisted plants breedingIn this technique, linkages are sought between DNA markers and agronomically important trait such as resistance to pathogens insects and nematodes etc. Instead of selecting for a trait the breeder can select for a marker that can be detected very easily in this selection scheme. The advent of molecular markers as a tool to aid selection has provided plant breeders with the opportunity to rapidly deliver superior genetic solutions to problems in agricultural production systems. However, a major constraint to the implementation of marker-assisted selection (MAS) in pragmatic breeding programs in the past has been the perceived high relative cost of MAS compared to conventional phenotypic selection. The point at which molecular markers are applied in a selection strategy can be critical to the effectiveness and cost efficiency of that strategy. Simulation studies have examined the potential role for MAS in breeding programmes (Hospital et al. 1997; Knapp 1998; Charmet et al. 1999; Moreau et al. 2000). These studies have shown that in some circumstances the adoption of MAShas the ability to improve selection efficiency over phenotypic selection alternatives. MOLECULAR MARKER Molecular marker- A molecular marker is a DNA sequence that is readily detected and whole inheritance can easily be monitored.Molecular marker are used to identify and tag desired gene. Different markers use for disease resistance is as follows

1.

RFLP (Restriction fragment length polymorphism )Developing RFLP probes Total DNA is digested with a methylation-sensitive enzyme (for example, PstI), thereby enriching the library for single- or low-copy expressed sequences (PstI clones are based on the suggestion that expressed genes are not methylated). The digested DNA is size-fractionated on a preparative agarose gel, and fragments ranging from 500 to 2000 bp are excised, eluted and cloned into a plasmid vector (for example, pUC18). Digests of the plasmids are screened to check for inserts. Southern blots of the inserts can be probed with total sheared DNA to select clones that hybridize to single- and low-copy sequences. The probes are screened for RFLPs using genomic DNA of different genotypes digested with restriction endonucleases. Typically, in species with moderate to high polymorphism rates, two to four restriction endonucleases are used such as EcoRI, EcoRV, and HindIII. In species with low polymorphism rates, additional restriction endonucleases can be tested to increase the chance of finding polymorphism. RAPD (Random amplified polymorphic DNA markers) SSRT (Simple sequence repeats or microsatellites AFLP (Amplified

2 3 4

fragment

length

polymorphism)

TISSUE CULTURE METHODSOMATIC HYBRIDIZATION Definition: A technique of fusing protoplasts from two contrasting genotypes for production of hybrids or cybrids which contain various mixtures of nuclear and/ or cytoplasmic genomes, respectively. (Chwala S.H. 1998) The protoplast, also known as naked plant cell refers to all the components of a plant cell excluding the cell wall. Hanstein introduced the term protoplast in 1880 to designate the living matter enclosed by plant cell membrane. The isolation of protoplasts from plant cells was first achieved by microsurgery on plasmolyzed cells by mechanical method (Klerckar, 1892). The true test of protoplast viability is the ability of protoplasts to undergo continued mitotic divisions and regenerate plants. The concentration of protoplasts in a given preparation can be determined by the use of hemocytometer The family Solanaceae and Brassiceae contains the most commonly used species for somatic hybridization. Both interspecific and intergeneric hybrids have been obtained. Many disease resistance genes viz. potato leaf roll virus, leaf blight, Verticillium, phytophthora have been transferred to Solonum tuberosum from other species where normal crossings would not be possible due to taxonomic or other barriers. Resistance to black-leg disease (Phoma lingam) has been found in B. nigra, B. jncea and B. carinata and after production of symmetric as well as asymmetric somatic hybrids between these gene donors and B. napus, resistant hybrids have been developed (Sjodin and Glimelius, 1989). Attempts were made to introduce tolerance in Brassica napus against Alternaria brassicae from Sinapsis alba (Primard et al, 1988) and beet cyst nematode from Raphanus sativus (Lelivelt et al. 1993). Resistance has been introduced in tomato against various diseases like TMV, spotted wilt virus, insect pests and also cold tolerance. Kobayeshi et al (196) reported the incorporation of disease resistance genes from wild species Solanum ochranthus a woody vine like tomato relative to L. esculentum. Hansen and Earle (1995) reported that black rot disease caused by Xanthomonas compesris is a a serious disease in cauliflower. The somatic hybrids were produced by protoplast fusion of B. oleracea with B. napus. Some of the examples of incorporation of resistance genes via protoplast fusion technique have been listed in Table.

Problems and Limitations of Somatic Hybridization Application of protoplast methodology requires efficient plant regeneration from protoplasts. Protoplasts from any two species can be fused. However, production of somatic hybrid plants has been limited to a few species. The lack of an efficient selection method for fused product is sometimes a major problem. The end-products after somatic hybridization are often unbalanced (sterile, misformed, and unstable) and are therefore not viable, especially if the fusion partners are taxonomically far apart. Regeneration products after somatic hybridization are often variable due to somaclonal variation, chromosome elimination, translocation, organelle segregation etc. It is never certain that a particular characteristic will be expressed after somatic hybridization. To achieve successful integration into a breeding program, somatic hybrids must be capable of sexual reproduction. All diverse intergeneric somatic hybrids reported are sterile and therefore have limited value for new variety development. It may be necessary to use back-fusion or embryo culture to produce gene combinations that are sufficiently stable to permit incorporation into a breeding program.

To transfer useful genes from a wild species to a cultivated crop, it is necessary to achieve intergeneric recombination or chromosome substitution between parental genomes (Chawla,2002).

Transgenics Transfer of genes between plant species has played an important role in crop improvement for many decades. Useful traits such as resistance to diseases, insects and pest have been transferred to crop varieties from noncultivated plants. The overall process of genetic transformation involves introduction, integration and expression of foreign gene in the recipient host plant. Plant that carry additional stably integrated, and expressed foreign genes transferred (transgenes) from other genetic sources are reffered to as transgenic plants. The capacity to introduce and express diverse foreign genes in plants was first described in tobacco by Agrobacterium mediated Horsch et al., 1984. And vectorless approach (Paszhkowski et al. 1984.) Methods to transform plants with DNA agrobacterium biolistics GENETIC ENGINEERING Genes expected to confer disease resistance are isolated, cloned and transferred into the crop in question. In case of viral pathogens, several transgenes have been evaluated, viz., virus coat protein gene, DNA copy of viral satellite RNA, defective viral genome, antisense constructs of critical viral genes, and ribozymes. Viral coat protein gene approach seems to be the most successful (Singh, 1998) A virus resistant transgenic variety of squash is in commercial cultivation in USA. In case of bacterial and fungal pathogens, resistance has been sought by expression of the following transgenes: 1) genes enoding insensitive target enzymes, 2) genes specifying toxin inactivation, 3) expression of antibacterial peptides, 4) expression of bacterial lysozymes, 5) genes specifying artificially programmed cell death, 6) expression of heterologous phytoalexins, 7) genes encoding ribsome inactivating proteins, 8) expression of heterologous thionins, 9) ectopic expression of pathogenesis related proteins and 10) ectopic expression of chitinases. In almost all the approaches, transgenic plants showed increased resistance to the concerned diseases. MERISTEM TIP CULTURE Cultivation of axillary or apical shoot meristems, particularly of shoot apical meristem is known as meristem culture. (Chawala H.S. 1998) Morel and Martin (1952) developed the technique of meristem culture for in vivo virus eradication of Dahlia. Meristem culture involves the development of an already existing shoot meristem and subsequently, the regeneration of adventitious roots from the developed shoots. It usually does not involve the regeneration of new shoot meristem When the objective is to free the stock from a virus, it is essential that the apical meristem should be excised along with a minimum of the surrounding tissue but when objective s vegetative propogation, the size of shoot tip used for culture is not important. Generally, explants taken for actively growing plants at the beginning of growing season are the most suitable. In practice shoot tip of upto 1 micro meter are used when the objective in virus elimination.

Advantages associated with meristem tip culture For production of disease free plants Meristem tip culture in generally followed where aim is to produce disease free plants. It has been demonstrated that the shoot apices of virus infected plants are frequently devoid of viral particles or contains very low viral concentrations. Though chemotherapeutic and physical agents have been used for production of virus free plants but with limited success. In vitro culture has becomes only effective technique to obtain virus free plants in potato, banana. Chrysanthemum, gladiolus, Pelargonium etc. from stock systemically infected not only with virus but with various other pathogens. Meristems are genetically stable and can be regenerated into pathogen free plants meristems have been identified as excellent material of germplasm preservation of crop species with seed borne viruses also. Disadvantages Facilities required are costly

Special skills are required to carry out the work Errors in maintenance of identify, introduction of an unknown pathogen, or appearance of an unobserved mutant may be multiplied to very high levels in short time. INSECT AND PEST RESISTANT CROPS

INTRODUCTION Insects are found in all types of environment and they occupy little more than two thirds of the known species of animals in the world. Insects affect human beings in a number of ways. Many of them fed on all kinds of plants including crop plants, forest trees, medicinal plants and weeds. They also infest the food and other stored products in godowns, bins, storage structures and packages causing huge amount of loss to the stored food and also deterioration of food quality. Insects inflict injury to plants and stored products either directly or indirectly in their attempts to secure food. Insects that cause less than 5 % damage are not considered as pests. The insects which cause damage between 5 10% are called minor pests and those that cause damage above 10% are considered as major pests. Insects that cause injury to plants and stored products are grouped into two major groups namely chewing insects and sucking insects. The former group chews off plant parts and swallow them thereby causing damage to the crops. Sucking insects pierce through the epidermis and suck the sap. Many of the sucking insects serve as vectors of plant diseases and also inject their salivary secretions containing toxins that cause severe damage to the crop. Introduction of high yielding varieties, expansion in irrigation facilities and indiscriminate use of increased rates of agrochemicals such as fertilizers and pesticides in recent years with a view to increase productivity has resulted in heavy crop losses due to insect pests in certain crops. This situation has risen mainly due to elimination of natural enemies, resurgence of pests, development of insecticide resistance and out-break of secondary pests. Distribution, nature of damage, life history of important key pests of crops and their management strategies are outlined hereunder: A NOVEL STRATEGY FOR PLANT PROTECTION: INDUCED RESISTANCE Most plant protection methods currently applied use toxic chemicals noxious to the environment for pathogen and pest control. Induced resistance exploiting natural defense machinery of plants could be proposed as an alternative, non-conventional and ecologically-friendly approach for plant protection. Its introduction into agricultural practice could minimize the scope of chemical control, thus contributing to the development of sustainable agriculture. Induced resistance can be defined as an increased expression of natural defence mechanisms of plants against various type of pathogens, provoked by a range of factors: pathogens causing hypersensitive necrotic reaction; avirulent or attenuated pathogenic strains; elicitors of pathogenic origin (glucans, proteins, lipids, etc.); abiotic elicitors, including synthetic harmless chemical products, such as 2,6-dichloroisonicotinic acid (INA), b-aminobutyric acid (BABA), benzothiadiazole (BTH), etc. Induced resistance, being based on the expression of latent genetic information present in plants, is not underlied by genome alterations (mutations, introgression of foreign genetic material), this enhancing its biological safety. What is induced resistance? When a plant is inoculated with a pathogen (primary inoculation), and after a time interval is subjected to a secondary (challenge) inoculation, reduced disease symptoms develop, i.e. the induced plant becomes more resistant than the normal, non-induced plant. Later, stimuli other than pathogens, such as some nontoxic chemicals, were found to be effective at inducing. resistance. Thus, the induced resistance can be defined as an increased expression of natural defense mechanisms of plants against different pathogens provoked by external factors of various type and manifested upon subsequent inoculation. The term induced resistance (IR) is used synonymously to acquired resistance (AR). Depending on the mode of its expression, induced resistance can be systemic (SAR) or local (LAR). Inducers of resistance Amultitude of factors are reported to induce resistance in plants: pathogens (fungi, bacteria, viruses) causing hypersensitive necrotic reaction (HR); avirulent and attenuated pathogenic strains; pests (insects, nematodes); elicitors of biotic origin; abiotic elicitors, i.e. chemical products, such as benzothiadiazole (BTH), _-aminobutyric acid (BABA), 2,6- dichloroisonicotinic acid (INA), salicylic acid, inorganic salts, etc. CHEMICAL INDUCERS The use of chemicals as inducers of resistance is an area of extensive work aiming at developing new compounds for disease control meeting the requirements for safe application in greenhouse and fields conditions, namely: no direct toxicity to pathogens; no toxicity to plants and animals; no negative effects on plant growth, development an yield; broad spectrum of defense; low loading amount; long lasting protection; low economical cost for farmers; good profit for producers Chemical inducers of plant resistance possess quite different mode of action as compared to fungicides and pesticides. The latter products have direct toxic effect on pathogens; are noxious to the environment; have narrow spectrum of defense; ensure shortly lasting protection; are economically costly. Thus, the application of chemical inducers of resistance is an exciting new perspective to supplement the classical chemical means of disease control by providing both effective and ecologically-friendly plant protection A large array of chemical products are shown to induce SAR in tobacco: salicylic acid, isonicotinic acid (INA), benzothiadiazole (BTH), _-aminobutyric acid (BABA) NaClO3, HgCl2, paraquat, polyacrylic acid, SiO2, etc. Chemically-induced SAR was found to be effective against fungi, bacteria and viruses. Mechanisms of SAR A cascade of molecular and biochemical events underlies the expression of SAR. It is initiated by perception of inducers (pathogens, chemicals) resulting in generation of signal molecules translocated at long distance, and switching on of diverse processes contributing to the development of the defense potential of plants realized upon secondary inoculation. Perception of inducers is effectuated through binding of pathogen-derived molecules (elicitors) or chemical products with receptor sites on plant membranes or cell walls. Generation and nature of signals, the mode of their translocation and SAR-mediating signal pathways may act simultaneously, thus providing an additive effect interactions are a matter of intensive research. Salicylic acid is commonly recognized as a signal molecule or a prerequisite for signal production in SAR; jasmonic acid and ethylene are involved in signalling upon expression of resistance induced by rhizobacteria and enter signal-transducing cascades involving MAPkinases Then interaction with gene promoters or other regulatory factors triggers the expression of the so-called SAR-genes. The term SAR-genes is used to collectively designate this family of nine genes whose expression is correlated

with the onset of SAR. Although not fully understood, induced resistance in plants opens new horizons in plant protection, being a promising tool for ecologically-friendly disease control and sustainable agriculture. It remains a challenge for both fundamental and applied research. PROTEINASE INHIBITOR Proteinase inhibitors (PIs) are anti-metabolic proteins, which interfere with the digestive process of insects. It is one of the important defence strategies existing in plants against predators. PIs inhibit the gut proteases of the insect which adversely affects the protein digestion in the gut and forces the insect to synthesize alternative protease to compensate for the inhibited activity. This leads to the deficiency of essential amino acid and exerts physiological stress on the insect leading to growth retardation. This mechanism of action reduces the possibility of developing resistance in insects and reduces crop damage. Transgenic plants expressing foreign insecticidal proteins have been produced for enhanced levels of insect resistance. PIs have been used because of their small size, abundance and stability. They are usually highly specific for a particular class of digestive enzymes of insects. Since the economically important classes of pests like Lepidoptera, Diptera and Coleoptera use serine and cysteine proteinases in their digestive system to degrade proteins in the ingested food. There are cowpea serine PIs (CpTI), potato serine PIs (PI-I and PI-II), sweet potato inhibitors, rice cysteine PIs, soybean Kunitz trypsin inhibitors, corn cystatins, mustard TI, tobacco PI and bean a-amylase inhibitors. Tobacco has been used as model plant in many studies to evaluate the efficacy of PIs as an insecticidal protein, followed by rice, potato, sweet potato, lettuce, tomato, oilseed rape, sunflower, petunia, birch, wheat, cauliflower, poplar pea and azuki bean. The first PI gene to be successfully transferred to tobacco, resulting in enhanced resistance against Manduca sexta was CpTI gene isolated from cowpea. In some cases, transgenically expresses PIs have not demonstrated any resistance against insects. Some insects have shown enough flexibility to switch proteinases composition in their guts to overcome the particular proteinase inhibitor expressed in the transgenic plants. Evidences have been presented to show that insects can adapt to the ingestion of PIs. Insects belonging to both Lepidoptera and Coleoptera can overexpress existing gut proteinases or induce the production of new types that are insensitive to the introduced PIs to overcome the deleterious effect of PI ingestion. This may be contributing for the decreased effectiveness of the PIs expressed in transgenic plants. In a recent study it was shown that high level expression of soybean trypsin inhibitor gene in transgenic tobacco plants failed to confer resistance against H. armigera. It is known that gut digestive enzymes are not the only targets affected by PIs; they can also affect the water balance, moulting and enzyme regulation of insects. To overcome this problem, an appropriate combination of PIs targeted to inhibit the complete spectrum of insect gut proteinases improves the stability of each and thus efficiently impairs digestion of dietary proteins in the insect gut. Additionally, a targeted statement of such a combination, specifically the differential temporal statement, will ensure exposure of the insect to different PIs in succession, forcing the insect to alter its mid-gut composition more than once leading to an additional physiological stress. - AMYLASE INHIBITORS -Amylases (a-1,4-glucan-4-glucanohydrolases, EC 3.2.1.1) constitute a family of endo-amylases that catalyse the hydrolysis of a-D-(1 4)glucan linkages in starch components, glycogen and other carbohydrates. The enzyme plays a key role in carbohydrate metabolism of microorganisms, plants and animals. Several insects, especially those similar to the seed weevils that feed on starchy seeds during larval and/or adult stages, depend on their a-amylases for survival. Six different a-amylase inhibitor classes, lectin-like, knottin-like, cereal-type, Kunitz-like, c-purothionin-like and thaumatin-like could be used in pest control. These classes of inhibitors show remarkable structural variety leading to different modes of inhibition and different specificity profiles against diverse a-amylases. Amylase inhibitors from wheat (WAAI) and common bean (BAAI) have been characterised. The common bean (Phaseolus vulgaris) contains a family of related seed proteins called phytohemagglutinin, arcelin and -amylase inhibitor (AI). AI forms a complex with certain insect amylases and is supposed to play a role in plant defense against insects. The introduction and expression of the bean -AI gene in pea confers resistance to the bruchid beetles. Transgenic Azuki bean carrying a-AI gene was resistant to three species of bruchids. Higgins and his group at CSIRO, Australia introduced -AI gene in an Indian genotype of chickpea (C-235) and derived significant protection against bruchids. However, bruchids such as Zabrotes can feed on plants producing a-AI because they possess a serine proteinase able to cleave some kinds of a-AI. It is therefore difficult to evaluate the long-term benefits of the expression of these genes in plants. PLANT LECTINS Lectins are carbohydrate-binding proteins that bind glycans of glycoproteins, glycolipids, or polysaccharides with high affinity. Plant lectins are classified mainly on the basis of their sugar binding properties. Most of the plant lectins are secretory proteins, meaning that they enter the secretory system and subsequently accumulate either in vacuoles or in the cell wall and intercellular spaces. toxic effects appear to be mediated through binding of the lectins to the midgut epithelial cells with consequent disruption of the cell function. The bound lectins may inhibit nutrient absorption or disrupt midgut cells by stimulating endocytosis of the lectins, and possibly other toxic metabolites present in the midgut. Lectins from snowdrop, pea, wheat, rice, castor, soybean, mungbean, garlic, sweet potato, tobacco, chickpea and groundnut have been isolated and characterised. Various lectins have been proved toxic towards members of the Coleoptera, Lepidoptera and Diptera. Lectins can be used to control sap-sucking insects belonging to the order Homoptera, which includes some of the most devastating pests. Bacillus thuringiensis Bacillus thuringiensis (or Bt) is a Gram-positive, soil-dwelling bacterium, commonly used as a biological pesticide; alternatively, the Cry toxin may be extracted and used as a pesticide. B. thuringiensis also occurs naturally in the gut of caterpillars of various types of moths and

butterflies, as well on leaf surfaces, aquatic environments, animal feces, insect rich environments, flour mills and grain storage facilities. During sporulation, many Bt strains produce crystal proteins (proteinaceous inclusions), called -endotoxins, that have insecticidal action. This has led to their use as insecticides, and more recently to genetically modified crops using Bt genes. Many crystal-producing Bt strains, though, do not have insecticidal properties. B. thuringiensis was first discovered in 1901 by Japanese biologist Shigetane Ishiwatari. In 1911, B. thuringiensis was rediscovered in Germany by Ernst Berliner, who isolated it as the cause of a disease called Schlaffsucht in flour moth caterpillars. In 1976, Robert A. Zakharyan reported the presence of a plasmid in a strain of B. thuringiensis and suggested the plasmid's involvement in endospore and crystal formation.B. thuringiensis is closely related to B.cereus, a soil bacterium, and B.anthracis, the cause of anthrax: the three organisms differ mainly in their plasmids. Like other members of the genus, all three are aerobes capable of producing endospores. Upon sporulation, B. thuringiensis forms crystals of proteinaceous insecticidal -endotoxins (called crystal proteins or Cry proteins), which are encoded by cry genes. In most strains of B. thuringiensis, the cry genes are located on a plasmid. Types of Bt crops:Bt cotton:- Bt cotton, a transgenic plant, produces an insect controlling protein Cry1A(c), the gene for which has been derived from the naturally occurring bacterium, Bacillus thuringiensis subsp. kurstaki (B.t.k.). The cotton hybrids containing Bt gene produces its own toxin for bollworm attack thus significantly reducing chemical insecticide use and providing a major benefit to cotton growers and the environment. Bt cotton contains the following three genes inserted via genetic engineering techniques: The Cry1Ac gene, which encodes for an insecticidal protein, Cry1Ac, derived from the common soil microbe Bacillus thuringiensis subsp. kurstaki (B.t.k.). The nptII gene, which encodes the selectable marker enzyme neomycin phosphotransferase II (NPTII), was used to identify transformed cells that contained the Cry1Ac protein. It served no other purpose and has no pesticide properties. The nptII gene is derived from the prokaryotic transposon Tn5. The aad gene which encodes the bacterial selectable marker enzyme 3(9) -O- aminoglycoside adenyltransferase (AAD) allowed for the selection of bacteria containing the PV-GHBK04 plasmid on media containing spectinomycin or streptomycin. The aad gene was isolated from transposon Tn7.

Bt corn:Bt maize has built-in protection against corn borers, achieved through modern biotechnology, where the Cry1Ab gene has been added. The Cry1Ab gene produces a Bt protein (Cry1Ab) which protects the plant from insect damage. This gene was derived from the common soil bacterium Bacillus thuringiensis, widely used as a biological control agent against various insect pests. Additionally, a marker gene (pat), has been added which gives the plant a tolerance to phosphinothricine, the active ingredient of glufosinate ammonium herbicides. This gene is derived from the soil bacterium Streptomyces viridochromogenes. The herbicide tolerance gene allowed selection of transformed plants in the development stage. Bt-11 insect-protected maize produces the Bt protein in its leaves, silks, stalks and ears throughout its life, enabling it to provide season-long protection against these devastating insect pests. Advantage:There are several advantages in expressing Bt toxins in transgenic Bt crops: The level of toxin expression can be very high thus delivering sufficient dosage to the pest. The toxin expression is contained within the plant system and hence only those insects that feed on the crop perish. The toxin expression can be modulated by using tissue-specific promoters, and replaces the use of synthetic pesticides in the environment. The latter observation has been well documented world-wide.

Disadvantage: Allergy Contamination of gene pool Antibiotics resistance gene Pest develop resistance against Bt crop Evolution of new pest

GENE PYRAMIDING Gene pyramiding, commonly called the stacking of genes through which the protective efficacy, spectrum of activity and the durability of resistance offered by the introduced genes can be greatly enhanced through careful design of packages of different genes that contain components which would act on quite different target insects. Gene pyramiding entails the simultaneous expression of more than one toxin in a transgenic plant. Gene pyramiding has been hailed as a lasting Bt resistance management strategy. However, a closer look at the strategy reveals that pyramiding was developed as a practical strategy to broaden the range of insect species that were not being adequately controlled by a single toxin as in the case of the single gene Bollgard Bt cotton variety. In a gene pyramiding scheme, strategy is to cumulate into a single genotype, genes that have been identified in multiple parents. In general, the gene pyramiding aims at the derivation of an ideal genotype that is homozygous for the favorable alleles at all n loci. The gene pyramiding scheme can be distinguished into two parts. The first part is called a pedigree, which aims at cumulating of all target genes in a single genotype called the root genotype. The second part is called the fixation step which aims at fixing the target genes into a homozygous state i.e. to derive the ideal genotype from the one single genotype. Each node of the tree is called an intermediate genotype and has two parents. Each of this intermediate genotype variety can resist. Moreover, pyramiding can also improve becomes a parent in the next cross. The intermediate genotypes are not just an arbitrary offspring of a given cross but it is a particular genotype selected from among the offspring in which all parental target genes are present. Although the pedigree step may be common, several different procedures can be used to undergo fixation in gene pyramiding. Generation of a population of doubled haploids from the root genotype is a possible procedure for the fixation steps. Here,

a population of gametes is obtained from the genotypes and their genetic material is doubled. This leads to a population of fully homozygous individuals, among which the ideotype can be found. Using this process, the ideal genotype can be obtained in just one additional generation after the root genotype is obtained. However, producing large population of doubled haploid is difficult and cumbersome in certain plant species. A possible alternative to this method is to self the root genotype directly to obtain the ideal genotype. MAS based gene pyramiding could facilitate in pyramiding of genes effectively into a single genetic background. The strategy of Bt gene pyramiding rests on three core assumptions. The first assumption is that insects resistant to only one toxin can be effectively controlled by a second toxin produced in the same plant. Eg. Bollgard II The second assumption is that strains resistant to two toxins with independent actions cannot emerge through selection pressure with one toxin alone. The third assumption is that a single gene will not confer resistance to two toxins that are immunologically distinct and that have different binding targets. Practical merits of gene pyramiding Second generation dual- Bt gene cottons Bollgard II (Cry 1Ac + Cry 2Ab) and WideStrike (Cry 1Ac + Cry1F) express two Bt endotoxins and were introduced in order to raise the level of control for H. zea, which was not satisfactorily controlled by the Cry 1Ac toxin alone. The Cry 1Ac and 2Ab toxins have different binding sites in the larval midgut and are considered to be a good combination to deploy in delaying resistance evolution. This is due to the fact that a species cannot easily evolve resistance to both toxins because that would require two simultaneous, independent mutations in genes encoding the receptors. Stack genes together no cross resistance eg. cry1Ac and cry2Ab, Cry1Ac resistant H. armigera will have broad spectrum cross resistance to all Cry1A toxins but lower Cross resistance to Cry2A. Rotation of two or three transgenic alternatively. Limitations of the Bt gene pyramiding strategy Diversity and plasticity of resistance genes: o The assumption that insects resistant to only one toxin can be effectively controlled by the second toxin produced by the plant fails to take full consideration of the enormous genetic plasticity in insect populations. Continuous exposure of insects to the active toxin: o Constitutive promoter in transgenic plants will express the toxin continuously at high levels for the duration of the growing season. o This creates some potential for cross resistance to pyramided genes, as it confers intense and uninterrupted selection pressure on the insects. Multiplicity of binding sites: o The assumption that strains resistant to two toxins with independent modes of action cannot evolve through selection with one toxin alone fails to account for the fact that one toxin can bind to several sites. INSECTICIDAL VIRUSES There are many viruses pathogenic to insect pests. These viruses are used in insect pest management programmes. Genomes of small viruses can be introduced into crop plants, which will synthesize the viral particles and acquire entomocidal property. For instance, H. armigera Stunt Virus (HaSV) is a tetravirus specific to lepidopteran insects and is very remotely related to viruses of plants and animals. HaSV is harmless to beneficial insects and the environment and its deployment in transgenic plants would not pose any risks46. A bio-prospecting approach is needed in India to identify such entomopathogenic viruses whose genomes can be manipulated in plants. PLANT SECONDARY METABOLITES Plants are equipped with an array of defense mechanisms to protect themselves against attack by herbivorous insects and microbial pathogens. Some of these defense mechanisms are preexisting, whereas others are only activated upon insect or pathogen invasion. Plant secondary metabolities are of 4 categories 1. chemicals of natural origin Alkaloids eg. Gramine Non-protein Amino Acids eg. L.DOPA Glucosinolates mustard oils Cyanogenic glycosides eg. Amygdalins Coumarins 2. Behavior modifying chemicals Repellents Antifeedants 3. Chemicals of plant defence - Biologically active substances Eg. Gossypol, Condensed tannins 4. Post infestational defence substances Antimicrobials Phytoallexins

Post infestation Process- signaling pathway:

Transcription factors such as, ERF 1 (Ethylene Response Factor 1), NPR 1 (nonexpressor of PR1 gene), GRX 480 (Glutaredoxin), MPK 4 (Map kinase 4) were found to function as an important signaling nodes that integrate signals from jasmonic acid, ABA and Ethylene in signaling pathways. Effective manipulation of these transcription factors responsible for the metabolic regulation at multiple steps in a pathway lead to more synthesis of target biochemicals. Effective manipulation of metabolic pathways involved in the production of plant secondary metabolite by introduction/elimination by antisense RNA technology of Enzyme encoding sequence and inhibiting competitive metabolic pathway to achieve metabolic flux towards higher production of particular molecule through antisense RNA and RNA interference technology, increase production of these compounds.