DEVELOPMENT AND MAINTENANCE OF INBRED LINES

BY
MUHAMMAD AWAIS FAROOQ

B.Sc. (Hons.) Agriculture 8th Semester Regd No. 2009-ag-2492

DEPARTMENT OF PLANT BREEDING AND GENETICS FACULTY OF AGRICULTURE UNIVERSITY OF AGRICULTURE FAISALABAD PAKISTAN 2013

CERTIFICATION Certified that the contents and format of the internship report entitled Development and Maintenance of Sunflower Inbred Lines during spring 2011 submitted by Mr. Muhammad Awais Farooq fulfills the partial requirement for the degree of B.S.c (Hons), Agriculture.

Internal Supervisor

_____________________ Professor Dr. Asif Ali Khan, Department of Plant Breeding and Genetics University of Agriculture Faisalabad

External Supervisor

_________________________ Dr. Muhammad Ayub Khan Principal Scientist Officer Oilseed Program, NARC.

Coordinator Oilseed Program

_____________________ Dr. Muhammad Amjad

____________________________________________________

Chairman Department of PBG

Dr. Abdus Salam Khan Professor

Dedicated To
TO MY LOVING FATHER AND MOTHER ONE WORD MOTHER COMPARE IT WITH NO ONE OTHER SO KIND AND SO FORGIVING THE FIRST WORD OF AN INFANT THE LAST WORD OF BRAVE PRAISE HER WHILE SHE IS LIVING FOR THERE IS NO TRUER RELATION THAN A MOTHER
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CONTENTS

ACKNOWLEDGEMENTS INTRODUCTON PARC NARC Oilseeds Research Program Sunflower Sunflower Inbred Development Program

REVIEW OF LITERATURE MATERIALS AND METHODS GENERAL INFORMATION ABOUT SUNFLOWER RESULTS AND DISCUSSION REFERENCES

ACKNOWLEDGEMENTS All praise to almighty Allah, the creator, the dominant, the most supreme, the self existing and all sustaining whose grace and mercy has enabled me to accomplish this research work by sacredness of Holy Prophet Muhammad (Peace be upon him) who is forever an entire source of guidance and knowledge for humanity. I am also thankful to Dr. Asif Ali Khan, my supervisor at University for making arrangements of my internship at NARC. I gratefully acknowledge the sincere efforts of Dr._Muhammad Amjad Coordinator Oilseed Program NARC, Islamabad for facilitating me in all possible ways to carry out my internship at Oilseed Research Program, NARC. I wish to express my sincere gratitude to my worthy supervisor Dr. Muhammad Ayub Khan Principal Scientific Officer, Mr. Saleem ud din, Research Fellow and all the field staff, Oil Seed Program, NARC, Islamabad for their continuous support, guidance, suggestions, encouragement and personal interest during this internship period. And I wish to express my real affection to my parent who supported me in every aspect of my life.

ABBREVIATIONS DFI PH NP SG g Ha Kg ORI PODB PARC PSC RARI SO RA SSO PSO CSO DG Days to Flower Initiation Plant Height Number of Plants Stem Girth Gram Hectare kilogram Oilseed Research Institute Pakistan Oilseed Development Board Pakistan Agriculture Research Council Punjab Seed Corporation Regional Agriculture Research Institute Scientific Officer Research Associate Senior Scientific Officer Principal Scientific Officer Chief Scientific Officer Director General

1.1.

IMPORTANCE OF INTERNSHIP PROGRAM

Internship program is compulsory requirement (with 20 credit hours) for the award of B.Sc. (Hons) Agriculture degree in which students of 8 th semester are sent to different research organization/institutions/stations to work with the scientists and gain practical knowledge in their respective field of specialization. This internship program provides opportunity to students in learning: How to conduct field experiments. Gain knowledge about the respective crop/field in depth. How to interact with supervisors and colleagues in the offices. What are the important parameters and how to record data on different parameters. Acquire skills in selfing and crossing techniques for crop improvement. Data compilation, analysis and report writing and presentation of research results.

Therefore, I understand that internship program is really a good opportunity for the students to gain practical knowledge in their respective field before entering in the practical life. As an essential requirement of internship program for the award of B.Sc. (Hons) Agriculture degree, I was placed by the Department of PBG in Oilseed Research Program, Crop Sciences Institute, National Agricultural Research Center, Islamabad as an internee. I joined Oilseed Research Program on march 12, 2013 and Coordinator Oilseeds, Dr. Muhammad Amjad, Coordinator advised me to work on sunflower. At that time sunflower research group has already planted sunflower experiments of summer season in last week of 25 February, 2013 and crop was at germination stage.

1.2

PAKISTAN AGRICULTURAL RESEARCH COUNCIL (PARC)

Pakistan (PARC),

Agricultural established

Research in 1981,

Council is an

autonomous apex body with the mandate to undertake aid, promote and coordinate

research at the federal and provincial levels in the country to provide science-based solutions for agricultural development.

Statutory Functions 1.3. Undertake aid, promote and coordinate research. Expedite utilization of research results. Train high level scientific manpower. Generate, acquire and disseminate agricultural information. Establish research library. NATIONAL AGRICULTURAL RESEARCH CENTRE (NARC) National Agricultural Research Centre (NARC) established in 1982, is the apex research institution of Pakistan Agricultural Research Council (PARC) at federal level. NARC serves as common platform for the scientists working in different federal and provincial agriculture research and academic institutions to jointly plan their research activities to avoid un-necessary duplication of

research activities. In particular, research on problems requiring sophisticated techniques and instruments like electron microscope, ultra centrifuges, molecular lab facilities and quality testing facilities are under taken at NARC supported by highly qualified and trained manpower. These facilities are also available to all the research scientists of the country. NARC field area is spread over 565 ha including office building and required infrastructure. Objectives: The center was established with the following objectives: To conduct high level basic/ applied research in crops, natural resources, livestock and agricultural production resources that require highly sophisticated equipments, trained manpower and can best be carried out at one place. To acquire, test and disseminate germplasm of various food grains, vegetables and fruit crops and to act as repository of seed and clonally propagated materials for food grains and horticultural crops. To develop appropriate agricultural machines and provide necessary technical support for mechanized farming. To establish a reference library and agricultural research information and documentation systems. To build in-service and pre-service manpower training facilities for the national agricultural research system.

Location NARC is located on the southern periphery of the Federal Capital and about 10 km from centre of the city. It is situated near Rawal Lake on the Park Road, Islamabad. Physical facilities in terms of experimental fields, laboratories, green houses, gene bank, library, documentation, auditorium, farm machinery and lab equipments, repair workshops, central stores, hostels, cafeteria, audiovisual studios, have also been established at NARC. The total manpower at NARC is 1241 with 149 Ph.D. 211 M.Phill/ M.Sc/M.A., 40 Graduates and 841 supporting staff.

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Organizational Structure The center is headed by the Director General, helped by the Directors of following different Institutes.

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1.4.

CROP SCIENCES INSTITUTE (CSI) Research Program Maize, Sorghum & Millet Research Program Oilseeds Research Program Pulses Research Program Sugar Crops Research Program Food Quality and Nutrition Program/Grain Quality Testing Lab Plant Physiology Program Research Program. Wheat Research Program Fodder Rice

The Crops Sciences Institute (CSI) has the following research programs:

1.5.

OILSEEDS RESEARCH PROGRAM Cooperative research program on oilseeds was started in 1977 with the primary objective of increasing the production of oilseed crops in Pakistan to achieve self sufficiency in edible oil. Oilseed Research Program on oilseeds coordinates at national level with the provincial research institutes and different stake holders in private sector for the research, development and promotion of oilseed crops in the country. Oilseed program is conducting research on different oilseed crops i.e. Rapeseed-mustard, Sunflower, Soybean, Groundnut, Sesame, Safflower and linseed.

Objectives of the program:

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 To coordinate with provincial research institutes and provide them germplasm and improved lines. Collection, evaluation, maintenance and multiplication of genetic material. Development of improved hybrids/varieties. Development of desirable cropping systems for oilseeds crops. Development of improved production technology for oilseeds crops. Identification and screening of diseases in oilseeds crops. Explore the potential areas for introduction oil seed crops.

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1.6. Oilseed Crops in Pakistan The oil crops in Pakistan are many, however, most of these are cultivated on small acreage. Rapeseed & mustard, sesame, linseed and castor bean are grown in this region since time immemorial and are called traditional crops. Sunflower, soybean, and safflower are called non-traditional. In addition, three oil bearing trees (coconut, olive and oil palm) also maintain their presence in some agro-ecological niches. Edible oil situation in Pakistan Pakistan is facing chronic deficiency of edible oil and its indigenous production is below the consumption level with a very wide gap between production and consumption. This gap is bridged through import of edible oil worth more than Rs.200 billion annually. Presently the domestic oilseed production only meets about 30-35 % of the total requirements. Cotton is the major source of edible oil in the country which contributes 6570 percent of the domestic production; sunflower is the second important oil crop contributing about 25% in domestic oilseed production, followed by rapeseed, mustard with 10 % contribution in year 2011-12 (Fig-1). For the last three decades, domestic edible oil production from all sources has grown at the rate of 2.56 % annually, whereas consumption is increasing at an annual rate of about 8 %. The indigenous edible oil production could not meet the demand of growing population. Therefore, rapid expansion of domestic oilseed production is only the solution of this problem. Sunflower Sunflower (Helianthus annuus L.), belongs to the family compositae. Its 2n chromosome number is 34. Sunflower is native of temperate regions of North America. During early 18th century, it was grown as an oil crop in Russia for the first time. There are two types of cultivated sunflower, i.e. Oil-type and Confectionary type

Its growth habit is annual with average plant height ranging from 1.5 to 2.5m while head diameter range is 15 to 30 cm. Sunflower is highly cross pollinated crop, honeybees being the major pollinators. The best temperature for its production is 20-25c. The leaves exhibit heliotropism they are directed East in the morning and turning to West in the evening. Its average 1000-seed weight ranges from 40-60 g with oil contents up to 55%. Its oil composition comprises of normally 25.1 % oleic, 66.2 % linoleic acids while protein contents ranged from 20-30 %. Sunflower is important source of vegetable oil at world level after soybean, oil palm and rapeseed, whereas, 2nd in Europe after rapeseed and also second in Pakistan after cotton seed. Area under sunflower (world) is about 30 mha, with a production of about 32 million tons. Major sunflower growing countries are Russia, Argentina, CIS, European Union (France, Spain, Germany, Romania, Bulgaria, and Yugoslavia), China, USA, and India. In Pakistan it is cultivated on area of 412,146 ha with the production of 603,894 tons.

Fig. 1. Trend in area and production of sunflower in Pakistan since 1970-71 to 2011-12.

. Major sunflower growing areas in the country are following. Punjab: Multan , Vehari , Lodhran , Bahawal Nagar, Bahawalpur , Rahim Yar Khan , D.G. Khan , Muzafar Garh and Rajan pur. Sindh: Thatta, Badin, Hyderabad, Dadu, Tando Allah Yar, Umar Kot and Nawab Shah Districts Khyber Pakhtun Khwa: Mardan, Sawabi, D.I.Khan, Bannu

Baluchistan Sibi, Quetta, Qallat, Nasirabad, Lasbela, Dera Murad, Jamali Average yield at farmers' fields is 1520 Kg/ha, while potential yield at progressive farmers field is 4000 Kg/ha. However, potential yield at research station/institute is > 4000 kg/ha. In Pakistan, sunflower was introduced during early sixties as oilseed crop, but now it is the second important source of vegetable oil after cottonseed. Its expansion since its introduction remained restricted due to the absence of systematic marketing mechanism, non-availability of quality seed and poor adaptability of imported hybrids. 1.6.1. Uses of sunflower oil

To date, sunflower oil has only been used as edible oil due to its higher price and limited supply in comparison to other oils. But due to its naturally high proportion of linoleic acid and advances in oil processing technology, the oil has advantages as a drying oil over linseed oil as it does not yellow with time. (The yellowing is due to the high linolenic acid content of the linseed oil). With the development of varieties high in oleic acid, and if a reduction in production costs can be achieved, sunflower oil could be used in the oleo chemical industry. However, there is limited production of non-oilseed varieties for use in confectionery and as bird feed. 1. Edible oil: Commercially available sunflower varieties/hybrids contain from 40 to 50%

oil in the seed. It is third largest source of vegetable oil worldwide. Sunflower oil is generally considered premium oil because of its light color, high level of unsaturated fatty acids and lack of linolenic acid, bland flavor and high smoke points. The primary fatty acids in the oil are oleic and linoneic (typically 90% unsaturated fatty acids), with the remainder consisting of palmitic and stearic saturated fatty acids. The primary use is as salad and cooking oil or in margarine. High oleic sunflower oil (80% oleic acids) is now available world-wide. It has expanded the

application of sunflower oils for frying purposes, tends to enhance shelf life of snacks, and could be used as an ingredient of infant formulas requiring stability. 2. Meal: Sunflower meal is higher in fiber, has a lower energy value and is lower in lysine

but higher in methionine. Protein percentage of sunflower meal ranges from 28% for nondehulled seed to 42% for completely dehulled seeds. The color of the meal ranges from grey to black, depending upon extraction processes and degree of dehulling. 3. Non-oil seed: The use of sunflower seed for bird feed or in human diets as a snack has

grown consistently over the past 15 years. Varieties used for non-oil seed purposes are characterized by a larger and require slightly different management practices. During processing, seed is divided into 1) larger seed for in-shell roasting, 2) medium for dehulling and 3) small for bird seed. Standards for different uses vary. 4. Forage: Sunflower can also be used as a silage crop. It can be used as a double crop after

early harvested small grains or vegetables, an emergency crop, or areas with a season too short to produce mature corn for silage. 5. Medicinal uses: i: The seeds of sunflower are used in treatment of bronchial and

pulmonary affections, coughs and colds, also in whooping cough. ii. A tincture of flowers and leaves has been recommended in treatment of bronchitis. iii. Sunflower oil like other oils, can retain moisture in the skin. It may also provide a protective barrier that resists infection in premature infants. 6. Industrial Application: There are several industrial applications of sunflower, e.g. used

in certain paints, varnishes and plastics because of good semi drying properties without color modification associated with oils high in linolenic acid, Sunflower oil is also used commonly in the manufacture of soaps and detergents .Sunflower oil is now used as bio-fuel, an alternate fuel source in diesel engines. Blends of sunflower oil and diesel fuel are expected to have greater potential then the burning of pure vegetable oil.

1.6.2. SUNFLOWER BREEDING PROGRAM AT NARC The Oilseed Research Program, NARC has strong and well established breeding program on sunflower. Breeding program was started in 1985 and developed more than 100 inbred lines (CMS, maintainer and restorer lines) and more than 1000 hybrid combinations. All the combinations were evaluated in preliminary yield trials at NARC and the promising ones were selected and included in multi-location trials (National Uniform Yield Trials (NUYT) for testing their adaptability under different environmental conditions. Since 2010, program was successful in developing four potential hybrids, having equal yield potential as imported hybrids (Hysun-33 from Australia and NK-S-278 from USA). One of the hybrids (Parsun-3) has been (recommended by variety evaluation committee) VEC on May 15, 2013 for approval.. 1.6.3. Development of Local sunflower Inbreds Sunflower breeding program at NARC has the following components Development of inbred lines Maintenance of inbred lines Development of new hybrid combinations Evaluation of Local & Exotic Inbred lines. Seed multiplication of parent inbred lines of potential hybrids Seed production of potential hybrids on pilot scale

Development of parent inbred lines used in the hybrid development takes 6-7 years for use in hybrid development. This involves a number of selfing and crossing steps to make the lines genetically pure and uniform. For any successful breeding program genetic variability of the crop is very important. Sunflower has narrow genetic base and the variability in the breeding material is also low. It is therefore, necessary to enhance the variability in available and new inbred lines by various means. As genetic variability is the

key to success in developing new varieties/hybrids. Therefore, at present more emphasis is to develop inbred lines with more genetic variation. 1.6.4. Internship in Oilseed Program When I joined sunflower breeding program, sunflower group had already planted following different experiments: 1) 2) 3) 4) National Uniform Yield Trials on Sunflower = (24 hybrids) Evaluation of local sunflower hybrids = (42 hybrids in 2 sets) Maintenance of sunflower inbred lines = (14 inbred lines) Development of hybrid combinations = 26

The above mentioned sunflower experiments were planted on 26th February 2013. H`owever, the experiment were badly damage due to hailstorm on March 14, 2013, therefore, fresh planting of the experiment was done on March 16 and 17, 2013. The Sunflower research group comprised of following scientists and field staff: S. No. 1. 2. 3. 4. 5. Name Dr. Muhammad Ayub Khan Dr. Muhammad Arshad Mr. Saleem-Ud-Din Mr. Abdul Ghaffar Mr. Muhammad Amin Designation Principal Scientific Officer Principal Scientific Officer Research Fellow Scientific Assistant Field man

REVIEW OF LITERATURE Venkanna, V. (2008) studied that twelve cytoplasmic male sterile lines belonging to four cytoplasmic male sterility (cms) sources were crossed with twelve inbreds to identify fertility restorer lines for each cms source. For the same source differing in line, the inbred behaved as either maintainer/restorer. Out of the twelve new inbred lines tested, fertility was restored in 9 inbreds for DRSF-114A (PEF), DRSF- 132A (PET-2) and DRSF-105A (PET-1), 11 inbreds for DRSF-116A (PEF) and DRSF-127A (CMSI), 4 inbreds for DRSF-115A (PEF), 5 inbreds for DRSF- 125A (CMSI) and DRSF-107A (PET-1), 6 inbreds for DRSF-124A (CMSI), DRSF-131A (PET-2) and 10 inbreds for DRSF-109A (PET-1). However, DRSF- 117A (PEF) was not restored by any of the inbreds tested. The study identified several effective restorers for newly developed cms sources for the first time in India, which can be exploited in developing highly heterotic hybrids possessing alternate cytoplasms.

Jayalakshmi, et al., (2001) studied that five inbred lines viz., IB19, IB29, IB32, IB41 and

IB49 were crossed with CMS 302A, CMS 234A, CMS 821A, CMS 850A and CMS 7-1A in a line x tester fashion to study their maintainer or restorer reaction. None of the inbreds could restore fertility in all the five CMS lines. IB29 and IB32 acted as restores for three CMS lines where as IB41 and 49 acted as maintainers for two CMS lines. The use of inbreds with restorer behaviour in future heterosis breeding programmes and those with maintainer behaviour in the development of new CMS lines through conversion has been suggested. Mohan and Seetharam., (2005) studied that genetic divergence was assessed in 137 inter specifically derived and known sunflower lines, 1 hybrid, 1 three way cross, and 1 open pollinated variety for 17 characters . The lines, irrespective of their pedigree, segregated into 6 clusters. Cluster I had maximum number (137) of lines. Highest intercluster distance was observed between clusters II and IV, followed by IV and VI, and IV and I. Selection of few lines from the clusters based on inter cluster distance and cluster mean values for hybridization is suggested. The divergence among the lines was

mainly due to five characters (accounting for 67.08%) given in the order of magnitude of contribution viz., head diameter, plant height, seed length, oil content, and days to 50% flowering. The limited divergence observed might be due to high levels of backcrossing to the recurrent parent (cultivated sunflower), which is normally in vogue in wide hybridization programs to obtain desirable agronomic background. However, it was difficult to obtain an optimized blend of genes from wild and cultivated forms leading to limited divergence in the resultant lines. Restricting to BC 1F2 generation for selecting superior introgressions and intercrossing might help in obtaining lines having higher heterogeneity and per se performance. Reddy et al., (2008) studied that fertility restoration in three diverse CMS sources of sunflower was studied using fifty inbreds (testers). While 22 inbreds maintained sterility of CMS PET 1 and CMS ARG 1, 28 inbreds restored their fertility. The third CMS line GIG 1, was maintained by all the inbreds indicating involvement of different gene(s). Most of the commercial sunflower hybrids are been produced using CMS PET 1. Now with the identification of restorers for CMS ARG 1, new more productive commercial hybrids can be produced. Efforts should be made to locate restorers for CMS GIG 1 for its utilization in production of sunflower hybrids. Horn and friedt, (1997) studied genetic diversity from 263 test crosses involving nine new CMS sources, i.e. ANL1, ANL2, MAX1, PEF1, PET2, ANN1, ANN2, ANNS and ANN4, five lines were selected as potential restorers for PEF1, PET2 and ANN4. In test crosses between all nine CMS sources and these five restorer lines evaluated in 2 years, seven fully restored hybrids could be identified. These hybrids, based on ANL1, ANL2, MAX1, PEF1, PET2, and ANN4, showed good agronomic performance for plant height, days to flowering, maturity and oil content. Segregation analyses of the F2 populations indicate that a single dominant restorer gene was sufficient to restore pollen production of hybrids based on ANL2, PEF1 and PET2. For restoration of ANN4, two dominant complementary genes are required. In restoration of fertility in the crosses of ANL1 and MAX1 investigated, two dominant genes are involved each of which on its own allows the production of fertile plants.

Sujatha et al., (2002) studied that the analysis of variance of 51 inbreds and 3 checks revealed significant differences for all 15 characters studied. The range of variation was maximum for percent autogamy followed by plant height and oil yield, while it was lowest in the case of days to 50 percent flowering and stem girth. The magnitude of variation was maximum for oil yield, 100-seed weight, head diameter, percent autogamy, husk percentage, seed density, plant height, seed test weight and oil content. Except for oil content, all other characters studied were least affected by environment showing a close correspondence between phenotypic co-variance (PCV) and genotypic co-variance (GCV). High heritability estimates were obtained for all the characters studied except for oil content, the value of which was moderate. High heritability coupled with the high genetic advance was recorded for plant height, percent autogamy, percent seed set and yield per plant. High heritability along with low genetic advance was observed for the number of leaves per plant, head diameter, stem girth, days to 50 percent flowering, 100-seed weight and oil yield, suggesting that these characters cannot be effectively improved by selection.

MATERIALS AND METHODS Development of Inbred lines The offspring of a cross-pollinating plant obtained as a result of repeated forced self-pollination. With repeated self-pollination the viability and yield of the offspring are decreased, while the homogeneity in respect to the genotype is increased. As a result of heterosis, hybrids from the cross breeding of the inbreds are substantially superior to the parent forms in vigor of development and seed yield. For this reason, inbreds are important in the production of hybrid seeds of field crops and vegetables. Varieties serve as the source material in the development of inbred lines. Succeeding generations are selected until the necessary homogeneity is attained, usually in the sixth generation. During selection, special attention is paid to the combining ability of inbred lines. The inbred lines that produce high yielding hybrids are selected. For the development of inbred lines following material may be used. 1) Adapted varietal populations, 2) Local varietal populations 3) Inter-varietal Hybrids, 4) Lineal Hybrid 5) Different gene pools. Method to work: In the first year of work 100 plants from source material are selfed to make S 0 generation. Since sunflower is prone to many diseases so it is better to test these lines for resistance before further experimentation. Promising lines are forwarded in successive generations on the basses of desired characters. The sunflower is an open pollinating plant species with a low percentage of autofertility. To obtain sufficient amount of seeds of inbred lines, it is necessary to help the pollinating by dabbing heads of individual plant with cotton wads. Seeds of S0 generation are sown next year in two rows, phenological observations are made. For developing inbred lines, attention should be paid to the following characters already in S1 generation:

1) Maturity 2) Plant height 3) Number of seeds per plant 4) Number of yield per plant in open pollination and selfing, 5) 1,000 seeds weight 6) Oil and protein contents in seeds 7) Resistance against diseases and pests (by inoculation method) It is equally important to know the manifestation of heterosis for the important agronomic characters and correlations for these characters between parents components and F1 progenies. Specific combining ability and general combining ability of the inbred lines should be tested. Combining ability testing may be started in 3 rd and 4th generation. Inbred lines are tested for general combining ability (GCA) by Poly cross method and top cross methods. Whereas, specific combining ability (SCA) is tested by the method of diallel crossing.

.3.1.

Data Recorded:

1. Days to flower initiation. 2. Plant height (cm) 3. Stem girth (mm) Procedure for data recording: 1. Days to flower initiation (DFI): Data on days to flower initiation were recorded from the date of planting till the date when 5 % of the buds per plot opened.

2. Plant height (PH): Data regarding plant height were recorded on randomly selected 10 plants from two central rows at physiological maturity in centimeters from the ground level to the attachment of head with the stem. 3. Stem girth: Data on stem girth (mm) were recorded 6 inches above the ground level of five randomly selected plants per plot and then mean values were calculation.

RESULTS AND DISCUSSION: STUDY OF QUANTITATIVE TRAITS OF SUNFLOWER

4.1. Development of inbred lines:

During the period of internship data were recorded on days to flowering initiation, plant height and stem girth. Information on days to flowering initiation is very important in sunflower breeding due to its single headed nature. Days to flower initiation It was observed that there was variation in days to flower initiation of different CMS lines ranged from 51 to 68 days. Whereas, days to flowering initiation was 52-69 days in the restorer lines. HA-64, HA-72, CMS-HA-76 and HA-80 took minimum days (51 days). Whereas, HA-71, HA-75, CMS-HA-77 and HA-77 were late in flower initiation and took maximum days (68) for flowering. Among restorer lines, some restorer lines were earlier in flowering except HEL-487 which was latest in flower initiation. RHP-46 also took more (68 days) for flower initiation.

Plant height (cm) Plant height is also an important agronomic trait in selection of sunflower inbred lines for high input response and better crop management. There was great variation in height of sunflower inbred lines ranging from 98- 203cm. it was observed that generally CMS and maintainer lines attained more height as compared to restorer lines with few exceptions. Among CMS lines, CMS-HA-70 and its maintainer line attained maximum height of (203cm and 195cm respectively. Whereas, CMS-HA-78 and HA-78 were shorter in stature. In restorer lines the maximum height (176cm) was recorded in HEL-487(MH) followed by HEL-203(MH) , RHP-72 and RHP-68 were shorter in stature with 100 and 101cm, respectively. Stem girth (mm) Stem girth or thickness is also important character because it protect crop from lodging. The inbred lines with stronger (thicker) stem can tolerate the lodging caused by extreme weather conditions.it was observed that restorer lines have relatively lesser girth as compared to CMS and maintainer lines. Stem girth recorded in different CMS lines varied between 13.86 and 32.66. HA-73 had maximum stem girth followed by CMS-HA-68 with 30.14mm. Among restorer lines, HEL-117 had maximum stem girth (30.05), followed by HEL-182 with 28.86mm. Minimum stem girth was recorded in RHP-42.

Table .2

Data on some agronomic traits of sunflower inbred lines used in

Hybrid Combinations Development

S.No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54

Inbred line HA-58 CMS-HA-58 HA-59 CMS-HA-59 HA-60 CMS-HA-60 HA-61 CMS-HA-61 HA-62 CMS-HA-62 HA-63 CMS-HA-63 HA-64 CMS-HA-64 HA-65 CMS-HA-65 HA-66 CMS-HA-66 HA-68 CMS-HA-68 HA-69 CMS-HA-69 HA-70 CMS-HA-70 HA-71 CMS-HA-71 HA-72 CMS-HA-72 HA-73 CMS-HA-73 HA-74 CMS-HA-74 HA-75 CMS-HA-75 HA-76 CMS-HA-76 HA-77 CMS-HA-77 HA-78 CMS-HA-78 HA-79 CMS-HA-79 HA-80 CMS-HA-80 HA-81 CMS-HA-81 HA-84 CMS-HA-84 HA-85 CMS-HA-85 HA-86 CMS-HA-86 HA-111 CMS-HA-111

DFI 62 58 60 64 56 56 57 64 61 61 56 57 51 52 57 56 56 57 60 60 60 57 67 66 68 65 52 51 56 54 66 63 68 64 66 61 68 68 54 56 61 59 61 65 59 61 52 57 59 59 53 58 54 56

PH (cm) 176 183 166 155 151 147 154 161 182 193 165 158 154 151 157 167 159 167 155 150 172 165 195 203 167 171 142 147 160 151 102 109 155 167 171 165 162 156 98 105 136 127 149 160 152 144 126 136 149 156 150 163 150 165

SG(mm) 18.46 15.30 24.02 15.98 13.86 13.92 21.50 23.56 24.94 21.73 24.10 20.08 19.56 22.69 16.07 29.62 9.50 29.88 28.36 30.14 27.29 20.15 25.26 18.33 19.88 18.72 16.26 23.70 32.66 26.11 17.06 17.11 19.00 25.62 18.31 14.45 24.32 18.47 23.44 19.72 19.85 18.13 23.85 24.15 26.05 18.12 15.81 19.80 25.14 22.59 28.86 21.21 19.79 24.85

SUMMARY
Development of Sunflower Inbred Lines is the main objective of sunflower breeding program because inbreeding lines with good combining ability are pre-requisite for successful hybrid development. Sunflower in Pakistan is planted with almost 100% imported hybrid seed on which huge amount of foreign exchange is spent. Therefore, efforts are being made at NARC to develop local hybrids with higher yield potential and other important agronomic traits. During spring 2013, parent lines (23 cytoplasmic male sterile lines, 23 maintainer lines and 18 restorer lines) were planted on 17th March 2013. There was significant variation in days to flowers initiation (51-69), plant height (98-203) and stem girth (13.86-32.66) in CMS lines and maintainer lines. Whereas, similar results were observed in the restorer lines in days to flowering initiation (52-69), plant height (100-176) and stem girth (11.23-30.05) respectively. CULTURAL OPERATIONS All the recommended cultural practices were done for better crop growth during the cropping season. Fertilizers were applied at a rate of 120 kg/ha Nitrogen, 60 kg/ha phosphorus and 60 kg/ha P2O5 were applied. Optimum plant population was achieved by maintaining proper plant to plant distance by manual thinning done after seedling emergence.

CONSTRAINTS IN PRODUCTION

1. Cold wet seasons (from establishing and harvesting point of view). 2. Lack of very early maturing cultivars of sunflower specifically selected for UK. 3. Disease, especially Sclerotinia sclerotiorum and Botrytis cinerea. 4. Birds in the short term, until a reasonable area of sunflower is established. Finches in particular can cause severe seed loss prior to harvest. MARKETS AND MARKET POTENTIAL

Anecdotal estimates suggest 60,000 ha could be grown in southern Britain, based upon land capacity. An edible semi-drying oil is obtained from the seed. Some varieties contain up to 45% oil. The oil is also used to make soap, candles, varnishes, paint etc, as well as for lighting and is said to be unrivalled as a lubricant. A blotting paper can be made from the seed receptacles and a high quality writing paper can be made from the inner stalk. The pith of the stems is one of the lightest substances known, having a specific gravity of 0.028. It has a wide range of applications, being used for purposes such as making life-saving appliances and slides for microscopes. The dried stems make an excellent fuel, the ash is rich in potassium. Both the dried stems and the empty seed receptacles are an excellent kindling. Fibre from the stem can be used to make paper and fine quality cloth. A yellow dye is obtainable from the flowers and a purple-black dye is obtainable from the seed of certain varieties. Sunflowers can be grown as a spring-sown green manure, they produce a good bulk of material, although root secretions from the plant can inhibit the growth of some plants.

Sunflower is perceived to be a drought tolerant crop as it roots deeply and extracts water at depths not reached by other crops. Sunflower is comparable to maize in many ways although it can extract water more efficiently in low-rainfall areas. The seedbed should be prepared so that a moist soil environment is available for germination and growth. The soil surface should be left as rough as possible to reduce the risk of soil erosion, drifting and blowing soil can seriously damage young seedlings. If the soil becomes compacted prior to planting reduced aeration and restricted water movement will occur, these conditions will increase the risk of downy mildew occurring. Breakdown of soil structure also reduces nutrient and water uptake and therefore yield. Sunflower has a wide potential sowing window. High yields may be produced from early plantings yet yields may be reduced by increased pest problems. Soil temperature should be a minimum of 7C for planting and around 10C for germination. Lower temperatures will increase the susceptibility of seedlings to diseases such as downy mildew. The main insect problems in sunflower crops include cutworms, sunflower bud moth, sunflower stem weevil, sunflower root weevil and the sunflower midge. Major disease problems include downy mildew, alternaria leaf and stem spot, septoria leaf spot, sclerotinia, botrytis, phoma, stem canker, verticillium wilt and nematode diseases. One of the major problematic weeds in the sunflower crop is wild oat, this can cause severe yield loss and should be treated if the threshold level is reached. Other potentially damaging problems include birds, rabbits, deer, mice, flooding and frost. Soils are often found to be deficient in nitrogen, phosphorus and sulphur. Potassium, calcium and magnesium are also frequently deficient in high-rainfall areas. Boron may also be required in some soils. Nitrogen applications of 50-75kg/ha are generally sufficient. The crop is generally harvested in September-October after a growing season of around 120 days (depending on summer temperatures, relative moisture distribution and fertility levels). Sunflowers should be grown in a suitable rotation to reduce the risk of weed, pest and disease attack, volunteers establishing, soil moisture depletion or phytotoxicity of the sunflower residue to the sunflower crop.

FERTILIZER RECOMMENDATIONS

Soil tests have been developed to estimate sunflowers potential response to fertilizer amendments. The most important factors in the fertilizer recommendations are the yield goal and the level of plant-available soil nutrients. In most climatic zones, predicting yield is impossible. Past yield records are a reasonable estimate of potential yield for the coming year. A yield goal for sunflower should be more optimistic than the average yield, and should approach the past maximum yield obtained by the grower on the same or a similar soil type. Nutrients not used by a crop in a dry growing season usually are not lost and can be used by the following crop. From an economic standpoint, having a yield goal that is somewhat high is much more beneficial for a grower than having a goal that is too low. A low yield goal in a good growing season easily can mean lost income of $30 to $40 per acre. In contrast, a high yield goal in a dry growing season will result in a loss of only $1to $2 in additional interest on the cost of unused nutrients since most of the nutrients will be available to the subsequent crop. The amount of nitrogen, phosphorus and potassium recommended for various sunflower yield goals and soil test levels are shown in Table 6. For yield goals not shown in the table, use the formulas at the base of the table. The data in this table are based on the amount of nitrate-nitrogen (NO3-N) in pounds per acre found in the top 2 feet of soil, the parts per million (ppm) of phosphorus (P) extracted from the top 6 inches of soil by the 0.5N sodium bicarbonate, and the ppm of potassium (K) extracted by neutral normal ammonium acetate in the top 6 inches of soil Other nutrients are not usually deficient for sunflower. On sandy slopes and hilltops, sulfur may be a problem; however, sulfur would not be expected to be deficient in higher organic matter, depressional soils. The sulfur soil test is a poor indicator of the probability of response to sulfur fertilizers. Sunflower has not been shown to be responsive to the application of other nutrients, including micronutrients in the state. FERTILIZER APPLICATION

Germinating sunflower seed is similar to corn in its reaction to seed-placed fertilizer. Application of more than 10 pounds per acre of nitrogen (N) plus potash (K2O) in a 30-

inch row will result in reduced stands or injured seedlings. Dry soil conditions can increase the severity of injury. In row widths narrower than 30 inches, rates of N plus K2O can be proportionally higher. For improved fertilizer rate flexibility, starter fertilizer should be placed in bands at least 2 to 3 inches from the seed row. Producers have several good reasons to apply nitrogen in the fall, such as availability of labor, soil conditions, etc. However, the general principle with respect to nitrogen application is: The longer the time period between application and plant use, the greater the possibility for N loss. In other words, use judgment in making a decision on time of N application. In the case of sandy soils, fall application of N is not recommended. In many instances, a side-dress application of when the sunflower plants are about 12 inches highmay be preferable. Phosphate and potash may be fall or spring applied before a tillage operation. These nutrients are not readily leached from soil because they form only slightly soluble compounds or attach to the soil. The phosphate and potash recommendations in are broadcast amounts. The recommendations for soil that tests very low and low in P and K can be reduced by one-third the amount in the table when applied in a band at seeding. In minimum or no-till systems, phosphate and potash may be applied in a deeper band to reduce the buildup of nutrients at the soil surface that occurs with these systems. However, most comparisons among deep, shallow and surface applications have shown little difference in crop response. WATER REQUIREMENTS FOR SUNFLOWER Sunflower has deep roots and extracts water from depths not reached by most other crops; thus it is perceived to be a drought-tolerant crop. Sunflower has an effective root depth around 4 feet, but can remove water from below this depth. Research on side-byside plots has shown that sunflower is capable of extracting more water than corn from an equal root zone volume. With its deep root system, it also can use nitrogen and other nutrients that leach below shallow-root crops; thus it is a good crop to have in a rotation. Seasonal water use by sunflower averages about 19 inches under irrigated conditions. Under dry land conditions, sunflower will use whatever stored soil moisture and rain that it receives during the growing season. When access to water is not limited, small grains use 2 to 3

inches less total water than sunflower during the growing season, whereas soybean water use is slightly greater. Corn uses 1 to 4 inches, and sugar beets use 2 to 6 inches more than sunflower, respectively, during the growing season. These total water use values are typical for non drought conditions in southeastern North Dakota. Small grains use the least total water since they have the fewest number of days from emergence to maturity. Sunflower and soybean have an intermediate number of days of active growth and corresponding relative water use. Corn ranks above sunflower in growth days and water use, while sugar beets rank highest in both categories. However, water use efficiency does vary among these crops. Comparative water use efficiency measured as grain (pounds per acre or lb/A) per inch of water used on three dry land sites and two years in eastern North Dakota was 119, 222, 307, 41, 218, 138, and 127 for sunflower, barley, grain corn, flax, pinto bean, soybean and wheat, respectively. These results indicated that corn had the highest water use efficiency, sunflower and wheat were intermediate and fl ax the lowest. (Source: M. Ennen. 1979. Sunflower water use in eastern North Dakota, M.S. thesis, North Dakota State University). Fertility has little influence on total water use, but as fertility increases, water use efficiency increases because yield increases. Yield performance has been shown to be a good indicator of water use efficiency of sunflower inbreds; higher yielding inbreds exhibit the highest water use efficiency. SOIL WATER MANAGEMENT Management practices that promote infiltration of water in the soil and limit evaporation from the soil generally will be beneficial for sunflower production in terms of available soil moisture. Leaving stubble during the winter to catch snow and minimum tillage are examples. Good weed control also conserves moisture for the crop. The use of postapplied and pre-emergence herbicides with no soil incorporation also conserves moisture when growing sunflower. Sunflower has the ability to exploit a large rooting volume for soil water. Fields for sunflower production should be selected from those with the greater water holding

capacity and soils without layers that may restrict roots. Water-holding capacity depends mainly on soil texture and soil depth. The loam, silt loam, clay loam and silty clay loam textures have the highest water-holding capacities. Water-holding capacity of the soils in any field can be obtained from county soil survey information available from local Natural Resources Conservation Service (NRCS) USDA offices. Sampling or probing for available soil moisture before planting also can help select fields for sunflower production. With other factors being equal, fields with the most stored soil moisture will have potential for higher yields. Where surface runoff can be reduced or snow entrapment increased by tillage or residue management, increases in stored soil moisture should occur and be beneficial to a deep-rooted crop such as sunflower.

TILLAGE, SEEDBED PREPARATION AND PLANTING Sunflower, like other crops, requires proper seed bed conditions for optimum plant establishment. Errors made at planting time may be compounded throughout the growing season. Seedbed preparation, soil tilth, planting date, planting depth, row width ,seed distribution and plant population should be nearly correct as conditions permit. TILLAGE AND SEEDBED PREPARATION Tillage traditionally has been used to control weeds and incorporate herbicides in preparation for planting. When tillage is used in low rainfall areas, producers must take care to control weeds while leaving as much of the previous crops residue intact as possible. Tillage never should occur when soils are too wet. Soils that are tilled when too wet and then dry will crust, turn lumpy and generally provide for poor seedbed conditions for germination and establishment. Maintaining a moist seedbed is important if producers expect to have uniform germination and emergence across the field germination and emergence will influence the need for and the effectiveness of future management practices. Excessive tillage should be avoided where tillage is used to prepare the seedbed or to incorporate preplant herbicides. Excessive tillage will break down soil structure, cause compaction and

crusting problems, reduce aeration, restrict water movement and provide conditions favorable for infection by downy mildew or other soil-borne diseases. Breakdown of soil structure also causes reduced nutrient and water uptake and reduces yield. Tillage and planting equipment is available to provide systems with varying levels of surface residue for Sunflower production. Production systems can range from conventional-till, where the quantity of surface residue covers less than 30 percent of the soil surface, to no-till, where the quantity of surface residue covers more than 60 percent of the soil surface. CONVENTIONAL-TILL PRODUCTION SYSTEMS Conventional-till production systems usually involve two or more tillage operations for weed control, incorporation of pre-emergence herbicides and incorporation of the previous crops residues. Pre-emergence herbicides may be incorporated with a tandem disk, chisel or sweep plows, disk harrow, long-tine harrow, rolling harrow or air seeders with sweeps in different sequences or combinations. Tillage sequences are determined by herbicide label requirements by the quantity of crop residue present at the beginning of the tillage operation, and by the seedbed requirements needed to match planting equipment capabilities. Conventional tillage systems, with or without preemergence or post-emergence herbicide, may include the option of row cultivation once or twice during the early growing season before the sunflower reaches a height too tall for cultivation. A rotary hoe or harrow can be used just before sunflower emergence and or at the V-4 to V6 development stage. Harrowing or rotary hoeing between emergence and the V-4 stage can result in injury or death of the sunflower plant. Depending on planting depth and stage of crop development, stand losses are generally less than 5 percent if the sunflower crop has at least two fully expanded leaves. Proper adjustment of the harrow or rotary hoe will maximize damage to the weeds and minimize injury to the sunflower crop.

REFERENCES Venkanna, V.*, Lokanadha Reddy, D. and Ranganatha, A.R.G. 2008. Identification of restorers and maintainers for different cms sources in sunflower using new inbreds. helia, 31, Nr. 49, p.p. 65-70. Jayalakshmi V., B. Narendra and S. Haritha., 2001. Maintainer/restorer identification for different cms lines in sunflower (helianthus annus l.). Agric. Sci. Digest, 21 (4) : 269 - 270, 2001. Mohan G.S. and A. Seetharam., 2005. Genetic divergence in lines of sunflower derived from interspecific hybridization. SABRAO Journal of Breeding and Genetics, 37(2) 77-84, 2005. C.V.C.M. Reddy, B. Sinha, A.V.V. Reddy and Y.R. Reddy, 2008. Maintenance of Male Sterility and Fertility Restoration in Different CMS Sources of Sunflower (Helianthus annuus L.). Asian Journal of Plant Sciences, 7: 762-766. Horn, R. and Friedt, W. (1997), Fertility restoration of new CMS sources in sunflower (Helianthus annum L.). Plant Breeding, 116: 317322. doi: 10.1111/j.1439-0523.1997.tb01005.x Sujatha, H.L., Chikkadevaiah and Nandini., 2002. Genetic variability study in sunflower inbreds. HELIA, 25, Nr. 37, p.p. 93-100, (2002).