CHAPTER 1 INTRODUCTION A.

Background of the Study Air pollution tolerance index (APTI) is used to determine plant species that are tolerant to air pollution (Agbaire, 2009 as cited by Seyyednjad and others, 2011). The plant species with APTI value that is less than 16 is considered sensitive and act as a bioindicator of air pollution; APTI value that ranges from 17 to 29 is considered intermediate and greater than 30 is tolerant to air pollution (Liu and Ding, 2008). The response of plants to air pollution at physiological and biochemical levels can be understood by analyzing the factors determining resistance and susceptibility. Studies show that air pollution has an impact on the ascorbic acid content, total chlorophyll content, leaf extract pH, and relative leaf water content of plants. For the reason that a single parameter may not provide a clear picture of pollution-induced changes, air pollution tolerance index (APTI) based on all four parameters has been used to identify the tolerance levels of plant species (Liu and Ding, 2008). Sensitivity and response of plants to air pollutants is variable. Using plants as an indicator of air pollution, is the possibility of synergistic action of pollutants (Lakshmi and others, 2009 as cited by Seyyednjad and others, 2011).

Several contributors agree that air pollution affects plant growth adeversely (Rao, 2006 as cited by Seyyednjad and others, 2011). In spite of the adverse affections of these pollutants, there are still plants that are tolerant to air pollution (Nivane and others, 2001 as cited by Seyyednjad and others, 2011). Plants play an important role in monitoring and maintaining ecological balance by actively participating in the cycling of nutrients and gases (Seyyednjad and others, 2011). Also, plants are an integral basis for all ecosystems and most likely to be affected by airborne pollution which are identified as the organisms with most potential to receive impacts from ambient air pollution. Also, the effects are most often apparent on the leaves which are usually the most abundant and most obvious primary receptors of large number of air pollutants. Biomonitoring of plants is an important tool to evaluate the impact of air pollution (Jyothi and Jaya, 2010).

Air pollution is a major problem arising mainly from industrialization (Odilara and others, 2006 as cited by Seyyednjad and others, 2011). Air pollution is the human introduction into the atmosphere of chemicals, particulate matter, or biological materials that cause harm or discomfort to humans and other living organism, or cause damage to the environment. Pollutants could be classified as either primary or secondary. Pollutants that are pumped into the atmosphere and directly pollute the air are primary pollutants while those that are formed in the air when primary pollutants react or interact are known as secondary pollutants (Agbaire and Esiefarienrhe, 2009 as cited by Seyyednjad and others, 2011).

Iloilo City has been industrializing for years; the populations in the area and the number of vehicles have been increasing, and new factories and commercial establishments were built. As an effect, the air around this area becomes more and more polluted. This study aimed to determine the air pollution tolerance index of three plant species (Ixora coccinea, Swietenia macrophylla, and Bougainvillea spectabilis) commonly found in the polluted and unpolluted areas of Iloilo City.

B.

Statement of the Problem Which among the three plant species (Ixora coccinea, Swietenia macrophylla, and Bougainvillea spectabilis) that are commonly found in the polluted and unpolluted areas of Iloilo City can be used as a bioindicator for air pollution?

C.

Objectives General objective: To determine the air pollution tolerance index of three plant species ( Ixora coccinea, Swietenia macrophylla, Bougainvillea spectabilis) commonly found in polluted and unpolluted areas of Iloilo City. Specific Objectives: To determine and calculate the relative leaf water content (RWC) of the three plant species (Ixora coccinea, Swietenia macrophylla, Bougainvillea
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spectabilis) that are commonly found in the polluted and unpolluted areas of Iloilo City. To determine and calculate the total chlorophyll content (T) of the three plant species (Ixora coccinea, Swietenia macrophylla, Bougainvillea spectabilis) that are commonly found in the polluted and unpolluted areas of Iloilo City. To determine and calculate the leaf extract pH of the three plant species (Ixora coccinea, Swietenia macrophylla, and Bougainvillea spectabilis) that are commonly found in the polluted and unpolluted areas of Iloilo City. To determine and calculate the ascorbic acid (AA) content of the three plant species (Ixora coccinea, Swietenia macrophylla, Bougainvillea spectabilis) that are commonly found in the polluted and unpolluted areas of Iloilo City.

D.

Significance of the Study

Air pollution is a major problem arising mainly from industrialization. Iloilo City has been industrializing for years and the air around this area becomes more and more polluted and; air pollution causes harm and discomfort to humans, to other living organisms and to the environment.

Plants play an important role in monitoring and maintaining ecological balance by actively participating in the cycling of nutrients and gases (Seyyednjad and others, 2011). Also, plants are an integral basis for all ecosystems and most likely to be affected by airborne pollution which are identified as the organisms with most potential to receive impacts from ambient air pollution. Also, the effects are most often apparent on the leaves which are usually the most abundant and most obvious primary receptors of large number of air pollutants. Biomonitoring of plants is an important tool to evaluate the impact of air pollution (Jyothi and Jaya, 2010). Studies show that air pollution has an impact on the ascorbic acid content, chlorophyll content, leaf extract pH, and relative water content of plants. For the reason that single parameter may not provide a clear picture of pollution-induced changes, air pollution tolerance index (APTI) based on all

four parameters has been used to identify the tolerance levels of plant species (Liu and Ding, 2008).

E.

Scope and Delimitation

This study focuses on the air pollution tolerance index of three plant species (Ixora coccinea, Swietenia macrophylla, Bougainvillea spectabilis) commonly found in the polluted and unpolluted areas of Iloilo City. The relative leaf water content (RWC), total chlorophyll content (T), leaf extract pH, and ascorbic acid (AA) content will be determined to calculate the air pollution tolerance index of the five species of plants.

F.

Definition of Terms Santan (Ixora coccinea) Santan is an erect and smooth ornamental shrub, growing to a height of 2 to 3 meters. Leaves are stalk less or on very short stalks, oblong, 5 to 9 centimeters long, heart-shaped or rounded at the base and blunt-tipped. Flowers are many, pink or red, and borne in terminal, stalk less or shortly stalked hairy cymes. Calyx teeth are short and pointed. Corolla-tube is slender, 2.5 centimeters long; lobes are spreading and oblong, about half the length of the tube. Fruit is reddish, almost round, about 5 millimeters in diameter. In this study, santan is a test organism taken from the polluted areas of Iloilo City. The air pollution tolerance index of santan was calculated to determine if it is sensitive to air pollution.

Mahogany (Swietenia macrophylla) Mahogany is a deciduous, erect tree growing to a height of 10 meters, with a heavy, dark-green, and dense crown. The trunk is more or less buttressed. Bark is dark gray and ridged. Young leaves when in the flush are pink, soon turning green. Leaves are alternate, smooth, compound, about 15 centimeters long, in 3 to 6 pairs, most often 5 pairs, of leaflets. Leaflets are inequilateral, ovate to oblong-ovate, 5 to 8 centimeters long
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and half as wide, pointed at the tip, broadly obtuse or rounded at the base. Flowers are greenish yellow, about 8 millimeters wide, born in axillary pannicles shorter than the foliage. Body of the fruit splits into five thick outer valves and five thinner inside valves. The outer valves fall off when ripe exposing closely packed seeds attached by the tips of their wings. Seeds are brownish, 5 to 7 centimeters long, with a broad and thin wing and a corky, thickened part containing the embryo. In this study, mahogany is a test organism taken from the polluted areas of Iloilo City. The air pollution tolerance index of mahogany was calculated to determine if it is sensitive to air pollution.

Bougainvillea (Bougainvillea spectabilis) Bougainvillea is a woody climber that can grow to a height of more than 10 meters, with large thorny stems and long drooping branches. The leaves are dark green, petioled, alternate, ovate, with entire margins, 6 to 10 centimeters long, broadest near the base. Thorns are the axils assist the plant in climbing. Flowers are in groups of threes, forming clusters at the terminal portion of the branches, each group subtended by three, broad, purplish, oblong-ovate and acuminate bracts, about 3 to 5 centimeters long. Flowers are small, each inserted on a bract, tubular, inflated midway through its length, of varying colors. In this study, bougainvillea is a test organism taken from the polluted areas of Iloilo City. The air pollution tolerance index of bougainvillea was calculated to determine if it is sensitive to air pollution.

Relative leaf water content (RWC) Relative water content is the amount of water a plant contains when it is incapable of taking in more water. This state is known as full saturation. A plant does not need to be in this state in order to survive but, knowing the percentage of water a plant is capable of holding is one way to determine if a plant is stressed. The high water content within a plant body will help to maintain its physiological balance under stress condition such as exposure to air pollution when the transpiration rates are usually high. High RWC favors
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drought resistance in plants. If the leaf transpiration rate reduces due to the air pollution, plant cannot live well due to losing its engine that pulls water up from the roots to supply photosynthesis (1%-2% of the total). Then, the plants neither bring minerals from the roots to leaf where biosynthesis occurs, nor cool the leaf (Liu and Ding, 2008). In this study, relative leaf water content (RWC) is one of the parameters that was determined to calculate the air pollution tolerance index of plants.

Total Chlorophyll Content (T) Total chlorophyll content is defined as the product of leaf chlorophyll content and total leaf area index. Depletion in chlorophyll immediately causes a decrease in productivity of plant and subsequently plant exhibits poor vigor. Therefore, plants maintaining their chlorophyll even under polluted environment are said to be tolerant ones (Singh and Verma, 2007). In this study, total chlorophyll content (T) is one of the parameters that was determined to calculate the air pollution tolerance index of plants.

Leaf extract pH Leaf extract pH is the pH of the extracts of the leaves of the plant. It can be determined using a pH meter calibrated with buffer solution of pH 4 and 9. High leaf extract pH may increase the efficiency of conversion from hexose sugar to AA, while low leaf extract pH showed good correlation with sensitivity to air pollution. Photosynthetic efficiency was noted strongly dependent on leaf pH. Photosynthesis is reduced in plants when the leaf pH was low (Liu and Ding, 2008). Plants with low lower pH are more susceptible to air pollution, while those with pH around 7 are more tolerant (Singh and Verma, 2007). In this study, leaf extract pH is one of the parameters that was determined to calculate the air pollution tolerance index of plants.

Ascorbic Acid (AA) content Ascorbic acid content is the amount of ascorbic acid present in the leaf of a plant. It is measured using iodine titration. Ascorbic acid plays a role in cell wall synthesis, defense, and cell division. It is a strong reducer and plays important roles in photosynthetic carbon fixation, with the reducing power directly proportional to its concentration (Liu and Ding, 2008). It also plays a significant role in light reaction of photosynthesis, activates defense mechanism, and under stress condition, it can replace water from light reaction. Ascorbic acid in plants has been shown to play an important role in pollution tolerance (Seyyednjad and others, 2011). In this study, ascorbic acid content is one of the parameters that was determined to calculate the air pollution tolerance index of plants.

Air Pollution Tolerance Index (APTI) The air pollution tolerance index (APTI) is used to determine the sensitivity of plant species when exposed to polluted air. It is used by landscapers to select plant species that are tolerant to air pollution. The plant species with APTI value that is less than 16 is considered sensitive and act as a bioindicator of air pollution; APTI value within 17 to 29 is considered intermediate and greater than 17 is tolerant to air pollution (Liu and Ding, 2008). In this study, air pollution tolerance index was used to determine the sensitivity and tolerance of plants to air pollution.

CHAPTER II REVIEW OF RELATED LITERATURE A. Air Pollution Tolerance Index (APTI) Sensitivity and response of plants to air pollutants is variable. Air pollution tolerance index is used to determine plant species that are tolerant to air pollution (Agbaire, 2009 as cited by Seyyednjad and others, 2011). The usefulness of evaluating APTI for the determination of tolerance as well as sensitiveness of plant species provided valuable informations for landscapers and greenbelt designers to select the sensitive as well as tolerant varieties of plant species (Jyothi and Jaya, 2010). The plant species with APTI value that is less than 16 is considered sensitive and act as a bioindicator of air pollution; APTI value within 17 to 29 is considered intermediate and greater than 30 is tolerant to air pollution (Liu and Ding, 2008). There are four parameters used to measure the APTI of plants: the relative leaf water content (RWC), the total chlorophyll content (T), the leaf extract pH, and the ascorbic acid (AA) content. These four parameters are calculated to attain the APTI values for each plant. For the reason that single parameter may not provide a clear picture of pollution-induced changes, air pollution tolerance index (APTI) based on all four parameters has been used to identify the tolerance levels of plant species (Liu and Ding, 2008).

Ascorbic acid plays a role in cell wall synthesis, defense, and cell division. It is also a strong reducer and plays important roles in photosynthetic carbon fixation, with the reducing power directly proportional to its concentration. So it has been given top priority and used as a multiplication factor in the formula. High pH may increase the efficiency of conversion from hexose sugar to AA, while low leaf extract pH showed good correlation with sensitivity to air pollution. Meanwhile, T, the TCh is also related to AA productivity and AA is concentrated mainly in chloroplasts. Photosynthetic efficiency was noted strongly dependent on leaf pH. Photosynthesis reduced in plants when the leaf pH was low. Thus, in the proposed APTI formula, P, the leaf extract pH and T, the TCh have been added together and then multiplied with AA content. A high water content within a plant body will help to maintain its physiological balance under stress condition such as exposure to air pollution when the transpiration rates are usually high. High RWC favors
8

drought resistance in plants. If the leaf transpiration rate reduces due to the air pollution, plant cannot live well due to losing its engine that pulls water up from the roots to supply photosynthesis (1%-2% of the total). Then, the plants neither bring minerals from the roots to leaf where biosynthesis occurs, nor cool the leaf. Therefore, the product of AA and sum of leaf extract pH and total chlorophyll is added with R, the RWC in the APTI formula (Liu and Ding, 2008).

A.1 Relative Leaf Water Content Relative water content is the amount of water a plant contains when it is incapable of taking in more water. This state is known as full saturation. A plant does not need to be in this state in order to survive but, knowing the percentage of water a plant is capable of holding is one way to determine if a plant is stressed. The high water content within a plant body will help to maintain its physiological balance under stress condition such as exposure to air pollution when the transpiration rates are usually high. High RWC favors drought resistance in plants. If the leaf transpiration rate reduces due to the air pollution, plant cannot live well due to losing its engine that pulls water up from the roots to supply photosynthesis (1%-2% of the total). Then, the plants neither bring minerals from the roots to leaf where biosynthesis occurs, nor cool the leaf (Liu and Ding, 2008). A.2 Total Chlorophyll Content (T) Total chlorophyll content is defined as the product of leaf chlorophyll content and total leaf area index. Depletion in chlorophyll immediately causes a decrease in productivity of plant and subsequently plant exhibits poor vigor. Therefore, plants maintaining their chlorophyll even under polluted environment are said to be tolerant ones (Singh and Verma, 2007). A.3 Leaf extract pH Leaf extract pH is the pH of the extracts of the leaves of the plant. It can be determined using a pH meter calibrated with buffer solution of pH 4 and 9. High leaf extract pH may increase the efficiency of conversion from hexose sugar
9

to AA, while low leaf extract pH showed good correlation with sensitivity to air pollution. Photosynthetic efficiency was noted strongly dependent on leaf pH. Photosynthesis is reduced in plants when the leaf pH was low (Liu and Ding, 2008). Plants with low lower pH are more susceptible to air pollution, while those with pH around 7 are more tolerant (Singh and Verma, 2007). A.4 Ascorbic Acid (AA) content Ascorbic acid content is the amount of ascorbic acid present in the leaf of a plant. It is measured using iodine titration. Ascorbic acid plays a role in cell wall synthesis, defense, and cell division. It is a strong reducer and plays important roles in photosynthetic carbon fixation, with the reducing power directly proportional to its concentration (Liu and Ding, 2008). It also plays a significant role in light reaction of photosynthesis, activates defense mechanism, and under stress condition, it can replace water from light reaction. Ascorbic acid in plants has been shown to play an important role in pollution tolerance (Seyyednjad and others, 2011).

B. Bioindicators A bioindicator is an organism or biological response that reveals the presence of the pollutants by the occurrence of typical symptoms or measurable responses, and is therefore more qualitative. These organisms (or communities of organisms) deliver information on alterations in the environment or the quantity of environmental pollutants by changing in one of the following ways: physiologically, chemically or behaviourally.

Bioindicator plant species are species with high sensitivity to ozone. This sensitivity is manifested in changes in physiological parameters, biomass reduction and the appearance of specific visible symptoms when exposed to high levels of ozone. According to the extent of these injuries, conclusions can be drawn regarding the presence (and, for some extent, the amount) of ozone in the atmosphere of the site where the plants are exposed to ambient air. In addition, ozone bioindicator plants are widely used in researches aiming to clarify plants response to the stressor, or, in general, to

10

oxidative stress. Reduction in biomass of sensitive clones as an effect of ozone is also a widely investigated indicative feature of biomonitoring plants.

C. Air Pollution Air pollution is a major problem arising mainly from industrialization (Odilara and others, 2006 as cited by Seyyednjad and others, 2011). Air pollution is the human introduction into the atmosphere of chemicals, particulate matter, or biological materials that cause harm or discomfort to humans or other living organism, or damage to the environment. Pollutants could be classified as either primary or secondary. Pollutants that are pumped into the atmosphere and directly pollute the air are primary pollutants while those that are formed in the air when primary pollutants react or interact are known as secondary pollutant (Agbaire and Esiefarienrhe, 2009 as cited by Seyyednjad and others, 2011).

C.1 Effects of Air Pollution on Humans The health effects of air pollution have been subject to intense study in recent years. Exposure to pollutants such as airborne particulate matter and ozone has been associated with increases in mortality and hospital admissions due to respiratory and cardiovascular disease. These effects have been found in shortterm studies, which relate day-to-day variations in air pollution and health, and long-term studies, which have followed cohorts of exposed individuals over time. Effects have been seen at very low levels of exposure, and it is unclear whether a threshold concentration exists for particulate matter and ozone below which no effects on health are likely.

A third cohort study (AHSMOG) found significant effects of particulate matter with a diameter of less than 10 m (PM10) on non-malignant respiratory deaths in men and women, and on lung-cancer mortality in male, non-smoking Seventh-Day Adventists.The effect on shortening life expectancy has been estimated at 12 years for realistic exposure contrasts, which is substantial compared with the effects of other lifestyle or environmental risk factors related
11

to mortality. Study show that the findings persist after inclusion of several more years of observation, with more consistent effects on lung cancer, in addition to non-malignant cardiopulmonary deaths. A Dutch study suggests that exposure to traffic-related air pollution is associated with cardiorespiratory deaths in much the same way as in the USA. For millions of people living in rural areas in developing countries, indoor pollution from the use of biomass fuels occurs at concentrations that are orders of magnitude higher than currently seen in the developed world. Deaths due to acute respiratory infections in children resulting from these exposures are estimated to be over 2 million per year (Brunekreef and Holgate, 2002).

C.2 Effects of Air Pollution on Plants Several contributors agree that air pollution affects plant growth adeversely (Rao, 2006 as cited by Seyyednjad and others, 2011). In spite of the adverse affections of these pollutants, there are still plants that are tolerant to air pollution (Nivane and others, 2001 as cited by Seyyednjad and others, 2011). Plants play an important role in monitoring and maintaining ecological balance by actively participating in the cycling of nutrients and gases (Seyyednjad and others, 2011). Also, plants are an integral basis for all ecosystems and most likely to be affected by airborne pollution which are identified as the organisms with most potential to receive impacts from ambient air pollution. The effects are most often apparent on the leaves which are usually the most abundant and most obvious primary receptors of large number of air pollutants. Biomonitoring of plants is an important tool to evaluate the impact of air pollution (Jyothi and Jaya, 2010). Studies show that air pollution has an impact on the ascorbic acid content, chlorophyll content, leaf extract pH, and relative water content of plants. The response of plants to air pollution at physiological and biochemical levels can be understood by analyzing the factors determining resistance and susceptibility (Seyyednjad and others, 2011).

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There are three principal air pollutants of major interest to agriculture sulfur dioxide, fluorine compounds, and smog (Thomas, 1963).

C.2.1 Sulfur Dioxide The effects of sulfur dioxide on plants are fairly well understood. The gas is absorbed into the mesophyll of the leaves through the stomata. Toxicity is due largely to the reducing properties of the gas. The limiting concentration that can be tolerated in the cells is about the same for many diverse species, including water plants. When this concentration is exceeded, the cells are first ion activated with or without plasmolysis, then killed. When extensive areas are killed, the tissues collapse and dry up, leaving a characteristic pattern of interveinal and marginal acute injury. Sulfate toxicity is a form of chronic injury manifested by white or brownish-red turgid areas on the leaf caused by the rupture of some cells or of chloroplasts within the cells (Thomas, 1963).

C.2.2 Fluorine Compounds Fluorides have great importance as air pollutants. Hydrogen fluoride and silicon tetrafluoride are toxic to some plants and fluorides particulates accumulate and build up concentrations on the inside and outside of the leaves causing marginal necrosis. Insoluble fluoride salts may be precipitated in the tissue, reducing the activity of the element. Fluorides can inhibit certain plant enzymes in very low concentrations (Thomas, 1963).

C.2.3 Smog Smog is a complex mixture of gases. There at least two types of smog: (1) mixture of coal smoke and fog with sulfur dioxide and (2) mixture of ozone and preoxidized organic compounds formed by photochemical reactions. Smog causes injury to the leaves giving it white or spotted collapsed areas on the upper surface. The chloroplasts of the
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plants exposed to smog disintegrate, plasmolysis then follows, and total dehydration of damaged cells results in "mummification" of the mesophyll tissue in the affected areas (Thomas, 1963).

D. Plant Species D.1 Santan (Ixora coccinea) Santan is an erect and smooth ornamental shrub, growing to a height of 2 to 3 meters. Leaves are stalkless or on very short stalks, oblong, 5 to 9 centimeters long, heart-shaped or rounded at the base and blunt-tipped. Flowers are many, pink or red, and borne in terminal, stalkless or shortly stalked, hairy cymes. Calyx teeth are short and pointed. Corolla-tube is slender, 2.5 centimeters long; lobes are spreading and oblong, about half the length of the tube. Fruit is reddish, almost round, about 5 millimeters in diameter. In this study, santan is a test organism taken from the polluted and unpolluted parts of Iloilo City. The air pollution tolerance index of santan was calculated to determine if it is sensitive or tolerant to air pollution.

D.2 Mahogany (Swietenia macrophylla) Mahogany is a deciduous, erect tree growing to a height of 10 meters, with a heavy, dark-green, and dense crown. The trunk is more or less buttressed. Bark is dark gray and ridged. Young leaves when in the flush are pink, soon turning green. Leaves are alternate, smooth, compound, about 15 centimeters long, in 3 to 6 pairs, most often 5 pairs, of leaflets. Leaflets are inequilateral, ovate to oblongovate, 5 to 8 centimeters long and half as wide, pointed at the tip, broadly obtuse or rounded at the base. Flowers are greenish yellow, about 8 millimeters wide, borne in axillary pannicles shorter than the foliage. Calyx is rimlike and the petals are oblong, less than 5 millimiters in length. Staminal tube is sligtly reddish, thick, and nearly as long as the corolla. Fruit is large, cylindrical, barrel-shaped, woody, grayish-brown, rough and less than 12 centimeters long. Body of the fruit
14

splits into five thick outer valves and five thinner inside valves. The outer valves fall off when ripe exposing closely packed seeds attached by the tips of their wings. Seeds are brownish, 5 to 7 centimeters long, with a broad and thin wing and a corky, thickened part containing the embryo. In this study, mahogany is a test organism taken from the polluted and unpolluted parts of Iloilo City. The air pollution tolerance index of mahogany was calculated to determine if it is sensitive or tolerant to air pollution.

D.3 Bougainvillea (Bougainvillea spectabilis) Bougainvillea is a woody climber that can grow to a height of more than 10 meters, with large thorny stems and long drooping branches. The leaves are dark green, petioled, alternate, ovate, with entire margins, 6 to 10 centimeters long, broadest near the base. Thorns are the axils assist the plant in climbing. Flowers are in groups of threes, forming clusters at the terminal portion of the branches, each group subtended by three, broad, purplish, oblong-ovate and acuminate bracts, about 3 to 5 centimeters long. Flowers are small, each inserted on a bract, tubular, inflated midway through its length, of varying colors. In this study, bougainvillea is a test organism taken from the unpolluted and polluted areas of Iloilo City. The air pollution tolerance index of bougainvillea was calculated to determine if it is sensitive or tolerant to air pollution.

E. Related Studies

E.1 Air Pollution Tolerance Indices of Some Plants Around Industrial Zone in South of Iran In the study of Seyyednjad and others (2011), the Air Pollution Tolerance Index of four plant species around petrochemical station in south west of Iran were examined.

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The samples were taken from the tree species in two places, polluted area and unpolluted area. Plants were randomly selected from the immediate vicinity of the station. Three replicates of fully matured leaves were used and immediately taken to the laboratory in the ice for analysis. The experiments were replicated three times for each biological factor. Air Pollution Tolerance Index was measured using the four parameters: relative leaf water content (RWC), total chlorophyll content (T), leaf extract pH, and ascorbic acid (AA) content.

The result showed order of tolerance in polluted area as E. camaldulensis (8/5) > A. lebbeck (8/1) > C. salignus (7/9) > P. juliflora (5/8) and in unpolluted area as E. camaldulensis (8/4) > A. lebbeck (6/7) > C. salignus (6/2) > P. juliflora (6/6). The results show that in cases that APTI increase from control site to polluted site improve the species tolerance to pollution stress.

All the samples are sensitive ones (1<APTI<16). However, P. juliflora is the most sensitive and its APTI showed reduction in polluted site as compared with control site. In conclusion, C. salignus, A. lebbeck, E. camaldulensis and P. juliflora can be used as bioindicators of air pollution.

E.2 Evaluation of air pollution tolerance index of selected plant species along roadsides in Thiruvananthapuram, Kerala In the study of Jyothi and Jaya (2010), a periodic evaluation of air pollution tolerance index (APTI) of selected tree species such as Polyalthia longifolia, (Sonner) Thw., Alstonia scholaris, R. Br., Mangifera indica, L., and shrubs Clerodendron infortunatum, L., Eupatorium odoratum, L., and Hyptis suaveolens, (L.) Poit., growing adjacent to the National Highway 47 passing through Thiruvananthapuram District which lies on the south-west coast of India, was carried out with a view to find out the air pollution tolerance as well as sensitivity of the plant species during different seasons.

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Among the trees in the roadside areas studied, Polyalthia longifolia, (Sonner) Thw., expressed highest APTI values and proved to be a tolerant variety and the others as sensitive species to air pollutants. In the case of shrubs, Clerodendron infortunatum, L., exhibited highest APTI values (7.34) and found to be more tolerant compared to the other two shrub species studied.

E.3 Variation in air pollution tolerance index of plants near a steel factory: Implications for landscape-plant species selection for industrial areas In the study of Liu and Ding (2008), twenty-three plant species growing near a Beijing steel factory, an air pollution point source, were collected during five dates from July 1 to October 16, 2001.

The data suggested that combining a variety of physiological parameters could give a more reliable result than those air pollution tolerance classifications based on a single biochemical parameter. Through the growing season, some species exhibited APTI variation related to changes in air temperature and water status of the plant. The results highlighted the need for APTI measurements to be conducted throughout the growing season, when evaluating pollution tolerance of individual species. Plant species tolerant or moderately tolerant to air pollution under a variety of environmental conditions include non-trees (shrub, herb, vine) such as Metaplexis japonica, Ampelopsis aconitifolia var. glabra, Rhamnus parvifolia, Ziziphus jujuba var. spinosa, Pharbitis purpurea, Vitex negundo, and trees including Broussonetia papyrifera, Robinia pseudoacacia, and Ailanthus altissima. The APTI of species indicated as an ideal candidate for landscape planting in the vicinity of polluting industry.

E.4 Evaluation of Air Pollution Tolerance Index (APTI) Of Some Selected Ornamental Shrubs in Enugu City, Nigeria In the study of Enete and Ogbonna in 2012, five species of ornamental shrubs that were growing along central business district (CBD) of Enugu Urban City of Nigeria were selected and evaluated for its air pollution tolerance index.
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Result indicates that ornamental shrubs had varied degree of tolerance index to air pollution. The air pollution tolerance index ranged from 10.60 to 14.32 with Ixora Red having the highest APTI value and Yellow Bush with lowest APTI value. The data suggested that ornamental shrubs growing in polluted environment often respond and show significant changes in their morphology, physiology and biochemistry. In this study, the orders of tolerance of ornamental shrubs are in this order: Ixora Red, Yellow Ficus, Masquerade Pine, Tuja Pine, and Yellow bush. The implication is that Ixora Red should be preferred where pollution appears high because of its ability to tolerate more pollutants. Ornamental shrubs like Yellow Bush showed are used as indicators of poor air quality.

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CHAPTER 3 METHODOLOGY A. Preliminary Parts A.1 Overview of the Study This study aimed to determine the air pollution tolerance index of three plant species (Ixora coccinea, Swietenia macrophylla, Bougainvillea spectabilis) commonly found in the polluted and unpolluted areas of Iloilo City. Specifically, this study aimed to: determine and calculate the relative leaf water content (RWC) of three plant species (Ixora coccinea, Swietenia macrophylla, Bougainvillea spectabilis) commonly found in the polluted and unpolluted areas of Iloilo City, determine and calculate the total chlorophyll content (T) of three plant species (Ixora coccinea, Swietenia macrophylla, Bougainvillea spectabilis) commonly found in the polluted and unpolluted areas of Iloilo City, determine and calculate the leaf extract pH of three plant species (Ixora coccinea, Swietenia macrophylla, Bougainvillea spectabilis) commonly found in the polluted and unpolluted areas of Iloilo City and, determine and calculate the ascorbic acid (AA) content of three plant species (Ixora coccinea, Swietenia macrophylla, Bougainvillea spectabilis) commonly found in the polluted and unpolluted areas of Iloilo City.

Three plant species (Ixora coccinea, Swietenia macrophylla, Bougainvillea spectabilis) were used in this study. Leaf samples were taken from the polluted and unpolluted parts of Iloilo City. Plants were randomly selected from the immediate vicinity of the station.

A.2 Time and Place of Study The samples for the study were taken from the polluted and unpolluted areas of Iloilo City. Laboratory works were conducted at Philippine Science High School Western Visayas Campus Research Laboratory. The study was conducted between the months of December 2012 and January 2013.

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A.3. Description of Study or Sampling site/s Leaf samples for the polluted area were taken from Philippine Science High School Western Visayas Campus (PSHSWVC). PSHSWVC is located in Brgy. Bitoon, Jaro, Iloilo City, a highly populated barangay, and near the floodway of Iloilo River, whose water goes directly to the Iloilo Strait. The school is approximately less than 7 kilometers away from the Panay Power BFO Plant which emits black smoke during operation. Also, cars and jeepneys would pass by at the school campus approximately every 10 minutes. On the other hand, leaf samples for the unpolluted area were taken from Barangay Balabag, Pavia, Iloilo. Pavia is a town located in the province of Iloilo. The location is surrounded by empty fields, some residential houses and approximately 1 kilometer away from the road.

B. Methodology

B.1 Preparation of Materials and Equipment B.1.1 Choosing of Plants and Collection of Leaves Three plant species (Ixora coccinea, Swietenia macrophylla, Bougainvillea spectabilis) were used in the experiment. Leaf samples were taken from the polluted and unpolluted parts of Iloilo City. Plants were randomly selected from the immediate vicinity of the station. Three replicates of fully matured leaves were used. The leaves from each plant were selected randomly. B.2 Preparation of Study Area The study was conducted on the polluted and unpolluted parts of Iloilo City. Plants were randomly selected from the immediate vicinity of the station. Laboratory works were held at Philippine Science High School Western Visayas Research Laboratory.

B.3 Measurement of the Air Pollution Tolerance Index (APTI) of the Plants

20

B.3.1 Measurement of the Total Relative Leaf Water Content (RWC) The Total Relative Leaf Water Content was determined according to the method described by Liu and Ding (2008). Fresh weight was obtained by weighing the fresh leaves. The leaves were immersed with water over night, were blotted dry and were weighed to get the turgid weight. The leaves were then dried overnight in an oven at 70C and were weighed again to obtain the dry weight. The Relative Leaf Water Content was calculated using the formula:
( ( ) )

Where: Fw = fresh weight Dw = dry weight Tw = turgid weight B.3.2 Measurement of the Total Chlorophyll Content (T) The Total Chlorophyll Content was determined using the method described by Lichtenthaler (1987). 0.2 g of fresh leaves was blended and was extracted with 10 mL of 80% acetone and left for 15 minutes. The liquid portion was decanted into another test-tube and centrifuged at 2,500 rpm for 3 minutes. The supernatant was collected and the absorbance was taken at 645 nm and 663nm using a spectrophotometer. Calculations were made using the formula: ( ) ( B.3.3 Measurement of the Leaf Extract pH Five grams of fresh leaves were homogenized in 10 mL deionized water. This was filtered and the pH of the leaf extract was determined after calibrating the pH meter with the buffer solution of pH 4 and 9. B.3.4 Measurement of the Ascorbic Acid (AA) Content ( ) )

21

2 g of potassium iodide was placed into a 100 mL beaker and 1.3 g of iodine crystals were added into the same beaker. A few mL of distilled water was added and swirled for a few minutes until the iodine crystals were dissolved. Iodine solution was transferred to a 1 L volumetric flask. A 1 L solution was made by adding distilled water up to the 1 L mark. 0.25 g of soluble starch was weighed and was added to 50 mL of near boiling water in a 100 mL conical flask. The solution was stirred and cooled before using. 100 g sample was cut into small pieces and was homogenized using a blender. 100 mL of distilled water was added while blending the sample. The grounded leaf was strained using a cheesecloth. The extracted solution was be made up to 100 mL with distilled water. The average volume of iodine solution used from the concordant titres and the moles of iodine reacting was calculated using the equation of the titration (below) to determine the number of moles of ascorbic acid reacting. ascorbic acid + I2 2 I + dehydroascorbic acid Then the concentration in mol L1 of ascorbic acid in the solution obtained from plant extract was calculated. Also, the concentration in g/mg Dw of ascorbic acid in the sample of plant was calculated. B.3.5 APTI determination The air pollution tolerance indices of the three plants was determined using the formula by Liu and Ding (2008). The formula for solving the APTI is given as: [ ( Where: A= Ascorbic acid content (mg/g Dw) T= Total Chlorophyll Content (mg/g Dw) P= pH of leaf extract R= Relative leaf water content (%)
22

C. Safe Handling of Chemicals Chemical containers were kept and labeled properly. The date when the chemical was received and was opened was recorded. Expiration dates and special storage conditions of chemicals were noted. All equipment were used for its designated purpose. D. Safety Procedures in the Work Area The laboratory was kept clean and free of equipment that are not relevant to the work. Work surfaces were disinfected after any spillage of possibly hazardous chemicals. Proper laboratory outfit was worn at all times while inside the laboratory. Close-toed shoes were worn inside the laboratory. Hands were washed before leaving the laboratory. E. Waste Disposal and After-care of the Work Area All materials and equipment that were used were cleaned after the experiment. The oven was thoroughly cleaned. The tables that were used for laboratory work was cleaned before and after use. After the laboratory work, the chemicals and apparatus should be returned to their designated areas. Spilled chemicals or broken glassware should be cleaned up, and disposed in appropriate waste containers. Soiled glassware should be cleaned at the laboratory sink or in laboratory dishwashers.

23

CHAPTER 4 RESULTS AND DISCUSSION This study aimed to determine the air pollution tolerance index (APTI) of three plant species (Ixora coccinea, Swietenia macrophylla, Bougainvillea spectabilis) commonly found in the polluted and unpolluted areas of Iloilo City. This was done by determining the total chlorophyll content (TCh), leaf extract pH, ascorbic acid content (AA), and relative leaf water content (RWC). Plant species with APTI value that is less than 16 is considered sensitive and act as a bioindicator of air pollution; APTI value within 17 to 30 is considered intermediate and greater than 30 is tolerant to air pollution (Liu and Ding, 2008). Three plant species (Ixora coccinea, Swietenia macrophylla, Bougainvillea spectabilis) were used in the experiment. Leaf samples were taken from Philippine Science High School Western Visayas Campus and Barangay Balabag, Pavia, Iloilo City. Plants were randomly selected from the immediate vicinity of the station. Leaf samples were brought to the laboratory for evaluation of total chlorophyll content (TCh), leaf extract pH, ascorbic acid content (AA), and relative leaf water content (RWC). Three trials were done during the months of December 2012 and January 2013. The APTI values of the plants were calculated using the formula of Liu and Ding (2008). A. Results Table1: Total chlorophyll content (TCh), Leaf extract pH, Ascorbic acid content (AA), Relative leaf water content (RWC), and air pollution tolerance index (APTI) of three plant species from Philippine Science High School Western Visayas Campus.
Philippine Science High School - Western Visayas (PSHSWV) Relative Leaf Water Content (%) Total Chlorophyll Content (mg/g Dw) Leaf Extract pH Ascorbic Acid Content (mg/g Dw) APTI

Santan (Ixora coccinea) Mahogany (Swietenia macrophylla) Bougainvillea (Bougainvillea spectabilis)

8.40

0.40

5.33

21.66

13.25

46.16

0.53

5.97

108.99

75.46

18.55

1.19

6.47

14.47

12.94

24

Table2: Total chlorophyll content (TCh), Leaf extract pH, Ascorbic acid content (AA), Relative leaf water content (RWC), and air pollution tolerance index (APTI) of three plant species from Barangay Balabag, Pavia, Iloilo
PAVIA Relative Leaf Water Content (%) Total Chlorophyll Content (mg/g Dw) Leaf Extract pH Ascorbic Acid Content (mg/g Dw) APTI

Santan (Ixora coccinea) Mahogany (Swietenia macrophylla) Bougainvillea (Bougainvillea spectabilis)

32.85

0.48

6.16

27.77

21.72

33.77

0.38

5.53

118.95

73.68

28.79

1.42

6.25

5.61

7.18

Table 1 and 2 shows the total chlorophyll content (TCh), leaf extract pH, ascorbic acid content (AA), relative leaf water content (RWC), and air pollution tolerance index of the plant species from Philippine Science High School Western Visayas Campus and Barangay Balabag, Pavia, Iloilo City as quantified during the months of December 2012 and January 2013. The air pollution tolerance indices of Mahogany (Swietenia macrophylla), Bougainvillea (Bougainvillea spectabilis) and Santan (Ixora coccinea) from PSHSWV are 75.46, 12.94 and 13.25, respectively. While, the air pollution tolerance indices of Mahogany (Swietenia macrophylla), Bougainvillea (Bougainvillea spectabilis) and Santan (Ixora coccinea) from Barangay Balabag, Pavia are 73.68, 7.18 and 21.72, respectively. Since, the APTI value of Mahogany (Swietenia macrophylla) is greater than 30 it shows that it is tolerant to air pollution while the APTI values of Bougainvillea (Bougainvillea spectabilis) and Santan (Ixora coccinea) indicates that these plant species are sensitive to air pollution since its APTI value is less than 16.
Table 3: T-Test results of the APTI values of plant species from PSHSWV and Brgy. Balabag, Pavia.

APTI value PSHSWV Pavia Bougainvillea PSHSWV Pavia Mahogany PSHSWV Pavia Santan

T-Test Results Significant Difference Significant Difference No Significant Difference

25

The air pollution tolerance indices of the three plant species from Philippine Science High School Western Visayas Campus and Barangay Balabag, Pavia, Iloilo were compared using the independent t-test. As shown in Table 3, the air pollution tolerance indices of Mahogany (Swietenia macrophylla) from the two study sites were found to be statistically insignificant. This proves the assumption that Mahogany is tolerant to air pollution while, the air pollution tolerance indices of Bougainvillea and Santan were found to be statistically significant. A significant difference between the APTI values would mean that a plant is sensitive to air pollution, making its values differ significantly when exposed to a more polluted air. This also proves the assumption that Bougainvillea and Santan are sensitive to air pollution. B. Discussion Air pollution tolerance index (APTI) is used to determine plant species that are tolerant to air pollution (Agbaire, 2009 as cited by Seyyednjad and others, 2011). The purpose of evaluating APTI for the determination of tolerance as well as sensitiveness of plant species could provide valuable information for landscapers and greenbelt designers to select the sensitive as well as tolerant varieties of plant species (Jyothi and Jaya, 2010). The plant species with APTI value that is less than 16 is considered sensitive and act as a bioindicator of air pollution; APTI value within 17 to 29 is considered intermediate and greater than 30 is tolerant to air pollution (Liu and Ding, 2008). There are four parameters used to measure the APTI of plants: the relative leaf water content (RWC), the total chlorophyll content (T), the leaf extract pH, and the ascorbic acid (AA) content. These four parameters are calculated to attain the APTI values for each plant. Ascorbic acid plays a role in cell wall synthesis, defense, and cell division. It plays important roles in photosynthetic carbon fixation, with the reducing power directly proportional to its concentration. So it has been used as a multiplication factor in the formula. High pH may increase the efficiency of conversion from hexose sugar to AA. Meanwhile, the TCh is also related to AA productivity and AA is concentrated mainly in chloroplasts. Thus, in the proposed APTI formula, the leaf extract pH and the TCh have been added together and then multiplied with AA content. The high water content within a plant body will help to maintain its physiological balance under stress condition such as exposure to air pollution. Therefore, the product of AA and sum of leaf extract pH and total chlorophyll is added with the RWC in the APTI formula (Liu and Ding, 2008).
26

The air pollution tolerance indices of Santan (Ixora coccinea) and Bougainvillea (Bougainvillea spectabilis) are less than 16 which mean that these plant species are sensitive to air pollution. The low APTI value of Santan is attributed to its low ascorbic acid content and to its low total chlorophyll content. The low chlorophyll content in plants would result to low ascorbic acid content (Liu and Ding, 2008). Although Bougainvillea has the highest total chlorophyll content of the three plant species, the leaf extract pH is slightly acidic which means that the pH is slightly low. Since high pH may increase the efficiency of conversion from hexose sugar to AA and the pH of bougainvillea is slightly low, this would be the possible reason why the ascorbic acid content of bougainvillea is also low. Ascorbic acid plays a role in cell wall synthesis, defense, and cell division. It is also a strong reducer and plays important roles in photosynthetic carbon fixation, with the reducing power directly proportional to its concentration (Liu and Ding, 2008). Bougainvillea also has the lowest relative leaf water content among the three plant species. The high water content within a plant body will help to maintain its physiological balance under stress condition such as exposure to air pollution when the transpiration rates are usually high. High RWC favors drought resistance in plants. If the leaf transpiration rate reduces due to the air pollution, plant cannot live well due to losing its engine that pulls water up from the roots to supply photosynthesis by 1%-2% of the total. Then, the plants neither bring minerals from the roots to leaf where biosynthesis occurs, nor cool the leaf (Liu and Ding, 2008). As a result, the APTI value of bougainvillea is low, making it a bioindicator for air pollution.

The air pollution tolerance index of Mahogany (Swietenia macrophylla) is greater than 30 which indicate that it is tolerant to air pollution. The high APTI value of this plant species is attributed to its high ascorbic acid (AA) content. Mahogany contains 118.95 g/mg Dw ascorbic acid. Ascorbic acid is a multiplication factor in the formula for solving APTI therefore; a high content of ascorbic acid in plant leaves would result to a high value of APTI for the plant. The values of the APTI acquired from the two different locations were compared using an independent t-test. The two study sites were Philippine Science High School Western Visayas Campus (PSHSWVC) and Brgy. Balabag, Pavia, Iloilo. PSHS-WVC is located in Brgy. Bitoon, Jaro, Iloilo City, a highly populated barangay, and near the floodway of Iloilo River, whose water goes directly to the Iloilo Strait. It is near the Panay Power BFO Plant which emits
27

black smoke during operation. Cars and jeepneys would pass by at the school campus from time to time. The other site is found at Brgy. Balabag, Pavia, Iloilo. Pavia is a town located in the province of Iloilo. The location is surrounded by empty fields, some residential houses and is far from the road. It was found out that the APTI values of Santan (Ixora coccinea) from the two study sites has a significant difference thus, proving that it is sensitive to air pollution and can act as a bioindicator. The APTI values of Bougainvillea (Bougainvillea spectabilis) from the two study sites were also found to be having a significant difference, thus making it acceptable to be used as a bioindicator for polluted air. A significant difference between the APTI values would mean that a plant is sensitive to air pollution, causing the values to differ significantly when exposed to a more polluted air. While, the APTI values of Mahogany (Swietenia macrophylla) from the two study sites are found to be having no significant difference and this proves the assumption that Mahogany (Swietenia macrophylla) is tolerant to air pollution and cannot act as a bioindicator for air pollution. The result of the current study becomes handy for future planning. The study also provides useful information for selecting tolerant species for streetscaping and microclimate modification. Species that are tolerant should be considered in advance for use; especially where air pollution is high conversely, species that are found to be sensitive should be utilized as bioindicators of urban air quality (Enete and Ogbonna, 2012). The analyses made agree with the study of Liu and Ding in 2008. APTI does not necessarily indicate if the location is polluted or not. The indication of air pollution is depends on whether the plant would survive on a location or not. Santan and Bougainvillea are both plants that are sensitive to the changes in the environment. If these plants would wilt or die after being planted on a certain location, we can most likely conclude that the air around is polluted. PSHS-WVC and Brgy. Balabag, Pavia are two places where Santan and Bougainvillea would tend to survive thus it is safe to say that the air around these places is not polluted.

28

CHAPTER 5 SUMMARY, CONCLUSIONS AND RECOMMENDATIONS This study sought to determine the air pollution tolerance index of three plant species (Ixora coccinea, Swietenia macrophylla, and Bougainvillea spectabilis) commonly found in the polluted and unpolluted areas of Iloilo City. This was done by determining the total chlorophyll content (TCh), leaf extract pH, ascorbic acid content (AA), and relative leaf water content (RWC). Plant species with APTI value that is less than 16 is considered sensitive and act as a bioindicator of air pollution; APTI value within 17 to 30 is considered intermediate and greater than 30 is tolerant to air pollution (Liu and Ding, 2008). A. Summary The findings of this study are summarized as follows: 1. Bougainvillea and Santan are considered to be sensitive to air pollution because the APTI values of these plants are less than 16. The APTI values of Bougainvillea and Santan from PSHSWV are 12.94 and 13.65, respectively. While the APTI values of Bougainvillea and Santan from Barangay Balabag, Pavia, Iloilo are 7.18 and 21.72. The APTI values of Bougainvillea and Santan from the two locations are statistically significant which proves that both Bougainvillea and Santan are sensitive to air pollution and can act as bioindicator. 2. Mahogany is tolerant to air pollution and it cannot act as bioindicator to air pollution. The APTI value of Mahogany from PSHSWV is 75.46 and the APTI value of Mahogany from Barangay Balabag, Pavia, Iloilo is 73.68. These values are statistically insignificant which proves that Mahogany is tolerant to air pollution.

B. Conclusions The air pollution tolerance index of Bougainvillea and Santan shows that it is sensitive to air pollution while the air pollution tolerance index of Mahogany indicates that it is tolerant to air pollution. Plant species that are found to be sensitive can act as
29

bioindicator to air pollution and plant species that are not sensitive to air pollution are highly discourage to be used as a bioindicator. C. Recommendations Future researchers on air pollution tolerance index are recommended to find the tolerance index of other plant species that are endemic in Iloilo City like Ipil-ipil, Banana, Mango and Coconut. To acquire a further result, the plant species must be taken from a more polluted area where it is more populated, urbanized, and industrialized. Researchers may consult at the Department of Environment and Natural Resources for the assessment handbook for polluted area. Furthermore, researchers must try to determine the air pollution tolerance index using multiple methods of measuring the four parameters: total chlorophyll content, relative leaf water content, leaf extract pH and ascorbic acid content.

30

LITERATURE CITED Brunekreef B and Holgate ST. 2002. Air pollution and health. 360: 1233-1242. Enete IC and Ogbonna CE. 2012. Evaluation of Air Pollution Tolerance Index (APTI) Of Some Selected Ornamental Shrubs in Enugu City, Nigeria. IOSR Journal of Environmental Science, Toxicology and Food Technology (IOSR-JESTFT). 1(2):2225. Jyothi SJ and Jaya DS. 2010. Evaluation of air pollution tolerance index of selected plant species along roadsides in Thiruvananthapuram, Kerala. Journal of Environmental Biology. 31: 379-386. Liu YJ and Ding H. 2008. Variation in Air Pollution Tolerance Index of Plants Near a Steel Factory: Implications for Landscape-Plant Species Selection for Industrial Areas. Wseas Trans. Environ. Dev. 4: 24-32. Pati P and Patra PK. 2012. Benthic Foraminiferal Responses to Coastal Pollution: A Review. International Journal of Geology, Earth and Environmental Sciences. 2(1):4256. Seyyednjad SM, Majdian K, Koochak H and Niknejad M. 2011. Air Pollution Tolerance Indices of Some Plants Around Industrial Zone in South of Iran. Asian Journal of Biological Sciences. 4(3): 300-305. Singh SK and Verma A. 2007. Phytoremediation of Air Pollutants: A Review. In: Environmental Bioremediation Technology, Singh SN and RD Tripathi (Eds.). Springer, Berlin Heidelberg. 293-314. Stuartxchange. 2011. Mahogany. Available: http://www.stuartxchange.org/Mahogany. html via the INTERNET. Accessed 2012 September 15. Stuartxchange. 2012. Bogambilya. Available: http://www.stuartxchange.org/Bogambilya. html via the INTERNET. Accessed 2012 September 15. Stuartxchange. 2012. Santan. Available: http://www.stuartxchange.org/Santan.html via the INTERNET. Accesed 2012 September 15. Thomas MD. 1963. Effects of Air Pollution on Plants. 233-278. University of Canterbury. Determination of Vitamin C Concentration by Titration. Available: http://www.outreach.canterbury.ac.nz/chemistry/documents/vitaminc_ iodine.pdf via the INTERNET. Accessed 2012 September 20.

31

APPENDIX A RAW DATA Actual Data Relative Leaf Water Content (RWC): PSHSWVC Fresh Weight Dry Weight Turgid Weight

1 Santan (Ixora coccinea) 2 3 1 Mahogany (Swietenia macrophylla) 2 3 1 Bougainvillea

(Bougainvillea spectabilis)

1.01 0.87 1.07 1.67 3.71 1.56 0.73 0.62 0.88

0.99 0.70 1.03 1.42 2.13 1.33 0.70 0.57 0.83

1.35 1.10 1.29 2.05 3.41 2.11 1.01 0.91 1.04

2 3

PAVIA Fresh Weight Dry Weight Turgid Weight

1 Santan (Ixora coccinea) 2 3 1 Mahogany (Swietenia macrophylla) 2 3 1 Bougainvillea

(Bougainvillea spectabilis)

0.76 0.99 0.91 1.06 2.18 1.85 1.88 1.72 1.80

0.60 0.91 0.71 0.92 1.80 1.53 1.77 1.67 0.93

0.99 1.33 1.23 1.45 2.82 2.38 2.38 2.27 2.38

2 3

32

Relative Leaf Water Content for Each Plant: PSHSWVC 1 Ixora coccinea Swietenia macrophylla Bougainvillea spectabilis PAVIA Ixora coccinea Swietenia macrophylla Bougainvillea spectabilis For Total Chlorophyll Content (T): PSHSWVC 1 Santan (Ixora coccinea) 2 3 1 Mahogany (Swietenia macrophylla) 2 3 1 Bougainvillea (Bougainvillea spectabilis) 3 PAVIA 1 Santan (Ixora coccinea) 2 3 1 Mahogany (Swietenia macrophylla) 2 3 1 Bougainvillea (Bougainvillea spectabilis) 2 3 2 1 41.03 26.42 18.03 5.56 39.68 9.68

2 4.25 69.30 14.71 2 19.05 37.25 8.33

3 15.38 29.49 31.25 3 38.46 37.65 60.00

Average 8.40 46.16 18.55 RWC 32.85 33.77 28.79

645 0.102 0.088 0.090 0.180 0.203 0.202 0.329 0.326 0.327 645 0.157 0.199 0.161 0.111 0.141 0.105 0.409 0.436 0.489

663 0.204 0.186 0.187 0.420 0.461 0.439 0.740 0.727 0.737 663 0.176 0.404 0.280 0.199 0.269 0.189 1.000 1.029 1.071

CT 3.70 3.27 3.32 7.00 7.80 7.60 12.58 12.42 12.52 CT 4.58 7.26 5.50 3.84 5.01 3.64 16.28 17.06 18.47

TCh 0.34 0.47 0.38 0.50 0.55 0.54 1.20 1.18 1.19 TCh 0.38 0.60 0.46 0.32 0.52 0.31 1.29 1.51 1.46
33

Average: PSHSWVC Ixora coccinea Swietenia macrophylla Bougainvillea spectabilis PAVIA Ixora coccinea Swietenia macrophylla Bougainvillea spectabilis Leaf Extract pH: PSHSWVC 1 Ixora coccinea 2 3 1 Swietenia macrophylla 2 3 1 Bougainvillea spectabilis 2 3 PAVIA 1 Ixora coccinea 2 3 1 Swietenia macrophylla 2 3

645 0.093 0.195 0.327 645 0.172 0.119 0.445

663 0.192 0.440 0.735 663 0.287 0.219 1.033

CT 3.43 7.47 12.51 CT 5.78 4.16 17.27

TCh 0.40 0.53 1.19 TCh 0.48 0.38 1.42

pH 5.1 5.3 5.6 6.6 5.0 6.3 7.3 6.1 6.0 pH 6.2 6.1 6.1 5.5 5.5 5.5
34

1 Bougainvillea spectabilis 2 3 Average: PSHSWVC Ixora coccinea Swietenia macrophylla Bougainvillea spectabilis PAVIA Ixora coccinea Swietenia macrophylla Bougainvillea spectabilis Ascorbic Acid Content (AA): PSHSWVC

6.2 6.3 6.3

pH 5.3 6.0 6.5 pH 6.1 5.5 6.3

Titrant (mL) 1 Ixora coccinea 2 3 1 Swietenia macrophylla 2 3 1 Bougainvillea spectabilis 2 3 PAVIA Titrant (mL) 1 Ixora coccinea 2 3 50 47 43 50 44.3 40 425 393.6 391.9 22.3 27.7 21

Ascorbic Acid Content 24.19 21.44 19.36 106.31 114.80 105.85 13.64 16.94 12.84 Ascorbic Acid Content 29.75 27.97 25.59

35

1 Swietenia macrophylla 2 3 1 Bougainvillea spectabilis 2 3 Average: PSHSWVC Ixora coccinea Swietenia macrophylla Bougainvillea spectabilis PAVIA Ixora coccinea Swietenia macrophylla Bougainvillea spectabilis

425 359.6 366.4 16 22 16.6

131.78 111.49 113.59 4.93 6.78 5.11

Titrant 44.77 403.50 23.67 Titrant 46.67 383.67 18.20

AA 21.66 108.99 14.47 AA 27.77 118.95 5.61

Air Pollution Tolerance Index (APTI): Relative PSHSWVC Total Leaf Water Leaf Chlorophyll Content Extract pH Content (T) (RWC) 1 5.56 0.34 5.1 Ixora coccinea 2 3 1 Swietenia macrophylla 2 3 1 Bougainvillea spectabilis 2 3 4.25 15.38 39.68 69.30 29.49 9.68 14.71 31.25 0.47 0.38 0.50 0.55 0.54 1.20 1.18 1.19 5.3 5.6 6.6 5.0 6.3 7.3 6.1 6.0

Ascorbic Acid Content (AA) 24.19 21.44 19.36 106.31 114.80 105.85 13.64 16.94 12.84

APTI 13.72 12.80 13.12 79.45 70.64 75.35 12.56 13.80 12.36

36

PAVIA

1 Ixora coccinea 2 3 1 Swietenia macrophylla 2 3 1 Bougainvillea spectabilis 2 3

Relative Leaf Water Content (RWC) 41.03 19.05 38.46 26.42 37.25 37.65 18.03 8.33 60.00

Total Chlorophyll Content (T) 0.38 0.60 0.46 0.32 0.52 0.31 1.29 1.51 1.46

Leaf Extract pH 6.2 6.1 6.1 5.5 5.5 5.5 6.2 6.3 6.3

Ascorbic Acid Content (AA) 29.75 27.97 25.59 131.78 111.49 113.59 4.93 6.78 5.11

APTI 23.80 20.67 20.74 79.73 71.29 69.87 5.50 6.09 9.96

Final Data
PSHSWVC Relative Leaf Water Content (%) Total Chlorophyll Content (mg/g Dw) Leaf Extract pH Ascorbic Acid Content (mg/g Dw) APTI

Santan (Ixora coccinea) Mahogany (Swietenia macrophylla) Bougainvillea (Bougainvillea spectabilis)

PAVIA

8.40

0.40

5.33

21.66

13.25

46.16

0.53

5.97

108.99

75.46

18.55

1.19
Total Chlorophyll Content (mg/g Dw)

6.47

14.47
Ascorbic Acid Content (mg/g Dw)

12.94

Relative Leaf Water Content (%)

Leaf Extract pH

APTI

Santan (Ixora coccinea) Mahogany (Swietenia macrophylla) Bougainvillea (Bougainvillea spectabilis)

32.85

0.48

6.16

27.77

21.72

33.77

0.38

5.53

118.95

73.68

28.79

1.42

6.25

5.61

7.18

37

APPENDIX B ANOVA TABLES Santan Group Statistics

Study site APTI value Pavia PSHSWV N 3 3 Mean 21.7367 13.2133 Std. Deviation 1.78724 .46705 Std. Error Mean 1.03186 .26965

Independent Samples Test

Levene's Test for Equality of Variances t-test for Equality of Means 95% Confidence Interval of the Difference F APTI value Equal variances assumed Equal variances not assumed 8.017 Sig. t df 4 2.272 Sig. (2Mean Std. Error tailed) Difference Difference .001 .010 8.52333 8.52333 Lower Upper

.047 7.992 7.992

1.06652 5.56221 11.48446 1.06652 4.42259 12.62408

Mahogany Group Statistics

Study site APTI value Pavia PSHSWV N 3 3 Mean 73.6300 75.1467 Std. Deviation 5.33025 4.40852 Std. Error Mean 3.07742 2.54526

Independent Samples Test

Levene's Test for Equality of Variances t-test for Equality of Means 95% Confidence Interval of the Difference F APTI value Equal variances assumed .356 Sig. t df 4 Sig. (2Mean Std. Error tailed) Difference Difference .723 -1.51667 Lower Upper 9.57136

.583 -.380

3.99360 -12.60469

Equal variances not assumed

-.380

3.864

.724

-1.51667

3.99360 -12.76032

9.72698

38

Bougainvillea Group Statistics

Study site APTI value Pavia PSHSW V N Mean 3 7.1833 3 12.9067 Std. Deviation 2.42269 .78009 Std. Error Mean 1.39874 .45038

Independent Samples Test

Levene's Test for Equality of Variances t-test for Equality of Means 95% Confidence Interval of the Difference F APTI value Equal variances assumed Equal variances not assumed 5.867 Sig. t df 4 2.410 Sig. (2Mean Std. Error tailed) Difference Difference .018 .044 -5.72333 -5.72333 Lower Upper

.073 -3.895 -3.895

1.46946 -9.80322 -1.64345 1.46946 -11.11869 -.32798

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APPENDIX C PLATES

Plate 1: Preparation of leaf samples

Plate 2: Weighing of leaves for relative leaf water content

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Plate 3: Immersing of leaves in water overnight for relative leaf water content.

Plate 4: Drying of leaves in an oven overnight for relative leaf water content.
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Plate 5: Weighing of leaves for ascorbic acid content.

Plate 6: Blending of leaves for ascorbic acid content, leaf extract pH and total chlorophyll content.
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Plate 7: Extraction of leaf extract for ascorbic acid content, leaf extract pH and total chlorophyll content.

Plate 8: Iodine solution as titrant for titration to determine ascorbic acid content.

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Plate 9: Titration for ascorbic acid content determination.

Plate 10: Measurement of leaf extracts pH

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Plate 11: Centrifugation of samples for total chlorophyll content.

Plate 12: Centrifugation of samples for total chlorophyll content.

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Plate 13: Samples for total chlorophyll content.

Plate 14: Measurement of total chlorophyll content using spectrophotometer.

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Plate 15: Measurement of total chlorophyll content using spectrophotometer.

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BUDGET Services Item RWC TCh AA pH Quantity (sample) 6 6 6 6 Price/qty (Php/sample) 0.00 0.00 0.00 0.00 Total Price (Php) 0.00 0.00 0.00 0.00

Laboratory Set-Up / Apparatus Item Quantity (piece) 100mL Beaker 1 Small test tubes 2 100mL Graduated Cylinder 1

Price/qty Php 180.00 Php 21.00 Php 348.00

Total Price Php 180.00 Php 42.00 Php 348.00

Transportation Item Family Car

Quantity 10 liters

Price/qty (Php/ride) Php 54.28/L

Total Price (Php) Php 542.80

Other Materials Item Gloves Tissue 1L Distilled Water 1.5L Distilled Water Small zip lock Big zip lock

Quantity 5 (pair) 1 (roll) 4 (bottle) 2 (bottle) 1 (box) 2 (box) Total Expenses

Price/qty Php 18.00 Php 14.25 Php 16.10 Php 35.00 Php 75.00 Php 150.00

Total Price Php 90.00 Php 14.25 Php 64.40 Php 70.00 Php 75.00 Php 300.00

Amount Services Laboratory Setup/Apparatus Transportation Other Materials Total Php 0.00 Php 570.00 Php 542.80 Php 613.65 Php 1726.45

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GANTT CHART Task Proposal Defense Preparation of Materials Collection of Leaves Measurement of RWC Measurement of pH Measurement of T Measurement of AA Calculation of APTI Recording of Data Interpretation of Data and Analysis Writing Chapters 4 and 5 Book Binding October November December January February

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