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AIR POLLUTION IMPACT ON REPRODUCTIVE BEHAVIOUR ON FEW TROPICAL TREES(AIR POLLUTION IMPACT ON FLOWER,FRUITS,SEED QUALITY & GERMINATION)

AIR POLLUTION IMPACT ON REPRODUCTIVE BEHAVIOUR ON FEW TROPICAL TREES(AIR POLLUTION IMPACT ON FLOWER,FRUITS,SEED QUALITY & GERMINATION)

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    Lambert Academic Publication(LAP),GERMANY Lambert Academic Publication(LAP),GERMANY Document Transcript

    • ContentsS.No. Particulars Page No. 1 Air Pollution Impact on Flowers 01-24 1.1 Introduction 1.2 Selection and Description of Test Species 1.2.1.1 Caesalpinia pulcherrima Swtz. (Swartz) 1.2.1.2 Cassia fistula Linn. 1.2.1.3 Cassia siamea Lamk. Syn. 1.2.1.4 Delonix regia Hook (Bojer ex Hook) 1.2.1.5 Peltophorum inerme Roxb. 1.2.2 Flowering Time 1.2.2.1 Air Pollutants 1.2.2.2 Sources of Air Pollution 1.2.2.3 Sources of Air Pollutants 1.2.2.4 Effects of Air Pollution 1.2.2.6 Need of Air Quality Monitoring 1.2.3 Soil Pollution 1.2.3.1 Types of Soil Pollution 1.2.3.2 Causes of Soil Pollution 1.2.3.3 Pollution Due to Urbanisation 1.2.3.4 Effects of Soil Pollution 1.2.3.5 Long Term Effects of Soil Pollution 1.2.3.6 Control of soil pollution 1.2.4 Noise Pollution 1.2.4.1 Sources of noise 1.2.4.2 Measures of noise 1.2.4.3 Effects of noise pollution 1.2.4.4 Causes and Effects of Noise Pollution 1.3 Results 1.3.1 Flowering Time 1.3.2 Floral Morphology 1.3.3 Flower Colour 1.3.4 Floral Biomass 1.3.5 Pollen Germination 1.3.6 Pollen Size 1.3.7 Pollen Tube Length 1.3.8 Pollen Viability 1.4 Discussion 1.4.1 Time of Flowering 1.4.2 Morphology of Flowers 1.4.3 Flower Colour 1.4.4 Floral Biomass 1.4.5 Pollen Characters i
    • 2 Air Pollution Impact on Fruits 25-36 2.1 Introduction 2.2 Experimental 2.2.1 Colour of Pods 2.2.2 Size of Pods 2.2.3 Weight of Pods 2.2.4 Seed Count 2.2.5 Seed Viability 2.3 Results 2.3.1 Colour of Pods 2.3.2 Size of Pods 2.3.3 Weight of Pods 2.3.4 Seed Count 2.3.5 Seed Viability 2.4 Discussion3 Air Pollution Impact on Seed Quality and Germination 37-47 3.1 Introduction 3.2 Experimental 3.2.1 Seed Colour 3.2.2 Seed Weight 3.2.3 Seed Density 3.2.4 Seed Soundness 3.2.5 Seed Germination 3.3 Results 3.3.1 Seed Colour 3.3.2 Seed Weight 3.3.3 Seed Density 3.3.4 Seed Soundness 3.3.5 Seed Germination 3.4 Discussion Acknowledgement 48 References 49-57 ii
    • Air Pollution Impact on Reproductive Behaviour of Few Tropical Trees Dr. Kishore Pawar, Dr. O.P.Joshi and Dr. Hema Swami Department of Environment Holkar Science College, Indore – 452 017- India Air Pollution Impact on Flowers1.1 Introduction To survive and do well, flowering plants have to reproduce themselves successfully. It is beneficial to the species if reproduction is carried out by sexual means, because this introduces greater variability in to the resulting offspring‟s, which in turns allow more opportunity for the species to evolve with its environment. The most colourful and spectacular aspects of plant growth are associated with development of flowers and fruits. Flower formation signifies a transition from vegetative to the reproductive phase of development. The shoot meristem is induced to develop sepals, petals, stamens and carpels instead of leaves. This transition can only occur at a particular time in the life of plants, which within certain limits, is determined genetically. Infect reproductive growth is certainly a complex process and physiologists have recognized a number of partial processes, which have been intensively studied. Ordinarily in thinking of reproductive growth, flower formation and fruit development come to mind. These events are obvious to the naked eye. However, each of these processes is the culmination of a number of other events, many of which are microscopic or submicroscopic. The reproductive growth is complex and encompasses a variety of anatomical, morphological, physiological and bio-chemical processes. After the plant attains the ripe to flower condition further progress towards flower initiation depends on the environment, both temperature and light are involved. Infect reproductive growth is certainly a complex processes, which have been intensively studied. 1
    • In this process two stages must be distinguished from each other, the induction of flowering and the differentiation of flowers and inflorescence. Everyone is familiar with flowers. Botanically flower is a modified shoot consisting of protective leaves, i.e. sepals and decorative coloured petals. These represent the parts of the flowers that is most familiar and indeed, generally thought of „the flower‟. Sepals and petals protect essential parts of the flowers, the male and female organs. The female part of the flower usually forms the central portion and it consists of one or more carpels, each of which contains one or more egg or ovules, mounted by a style and stigma. The stigma is the receptive surface on which pollen grains can land and grow, while the style is simply its stalk. The male parts are usually found in a ring around the central female parts, and they consist of pollen bearing stamens. Flowers may be produced singly or in- group known as inflorescence. The purpose of these aggregations normally seems to be an aid in attracting potential pollinators. During postembryonic development in higher plants, the shoot apex undergoes three discernible phases - juvenile vegetative, adult and reproductive. The transition from the juvenile to adult phase is usually gradual and involves subtle changes in shoot morphology and physiology (Poethig 1990). The intermediate developmental patterns are common during the transition from vegetative to reproductive stages. Infect, differentiation of the reproductive organ is preceded by formation of sepals and petals. That has a combination of vegetative and non-vegetative characters (Shrivastava and Iqbal 1994). For flowering the size of the shoot is more important than its age. In several species, shoot undergoes flowering on reaching a certain stage of development (Robinson and Warening 1969). The regulatory mechanism ensures that the plant does not flower until it has attained the requisite size. This holds true even in plants requiring a specific day length or chilling.1.2. Selection and Description of Test Species The five tree species selected for the present study belongs to family- Fabaceae. These trees are of good ornamental value. They are planted on roadside, in gardens and even in home gardens. They give a good colour effect; attract birds, bees and butterflies, which pollinate them. They are important component of 2
    • urban ecosystem and presently facing threat due to harmful and toxic effect of urban air pollutants. A brief taxonomic description of these plants is presented below:1.2.1.1 Caesalpinia pulcherrima Swtz. (Swartz) A glabrous shrub or a small tree unarmed or with a few weak prickles, cultivated in gardens, generally throughout India (Plate 1.1). It is commonly known as Shankhasur, Gultora, Chhoti-gulmohar and Krishna chura. Leaves- 15-30 cm. Long, alternate, pinnae 6-8, leaflets, 8-12 in pairs, sessile and oblong. Flowers- Scarlet yellow or red in elongate auxiliary and terminal racemes. Total five petals sub-equal, transversely oblong. Stamens- 10, free, filament long, petaloid. Pods- Oblong and flat, glabrescent, narrower and thinner than those of any of the genus. Seeds- 8-10 obviate – oblong and glabrous. Flowering- Nearly throughout the year. Plate 1.1: Caesalpinia pulcherrima in flowering at Low Pollution Area1.2.1.2 Cassia fistula Linn. A very handsome tree 20-30 feet high, trunk straight, bark smooth and pale gray when young, rough and dark-brown, when old, branches spreading, slender (Plate 1.2). This is a well-recognized avenue tree, occasionally found in deciduous forest also, commonly known as Amaltas. 3
    • Leaves- 9-16 inches long main rachis pubescent, stipules minute, linear oblong,obtuse, pubescent.Leaflets- 4-8 pairs, ovate or ovate-oblong, acute, bright green, glabrous andsilvery-pubescent beneath when young, the midrib densely pubescent at theunderside, base cuneate. Plate 1.2: Cassia fistula with developing pods showing flowers in inset.Flowers- In racemes 12-20 inches long, pedicels 1½ - 2 ¼ inches long, slender,pubescent and glabrous.Sepals- 5, pubescent, oblong.Petals- 5, sub-equal obovate, shortly clawed, veined.Stamens- 10, the 3 longest stamens are much curved and bear large, oblongcurved anthers, the 4 median stamens are straight and 3 remaining are very shortand erect staminode, dehiscing longitudinally by pores.Pods- 2-3 feet long, 1-3/4 inches in diameter, pendulous, cylindrical, nearlystraight, smooth, shining, brown-black, not torulose, indehiscent with numerous(40-100) horizontal seeds immersed in a dark coloured sweetish pulp, andcompletely separated by transverse partition.Flowering- April-June.Fruiting- Persisting throughout the year. 4
    • 1.2.1.3 Cassia siamea Lamk. Syn. Evergreen tree of moderate size having nearly smooth, gray bark marked with slight longitudinally fissures (Plate 1.3). The Cassia siamea is a native of South India and Burma. It is now grown throughout the India planted on roadside and in gardens for its shade and showy flowers. Its dark green, glossy leaves are divided into two rows of narrow, pointed leaflet arranged in opposite pairs on the slender midrib. Plate 1.3: Cassia siamea with flowers and fruits at Low Pollution Area. Leaves- Peripinnate about 12 inches long, leaflets 12 to 20, elliptical-oblong, mucronate, glabrous. Flowers- Yellow grow in large, open clusters at the ends of the branches about 1 ¼ inches, each of the flower having five almost equal petals and perfect seven stamens nearly unequal that produce pollen, the remaining three stamens being wanting, or small and sterile. Pods- The flat pods are purplish or brown, when ripe and contain a number of seeds. When young, they are soft, ribbon-like, minutely velvety, 6 to 9 inches long. Flowering– Throughout the year, maximum flush is observed in October. Fruiting- April and throughout the year.1.2.1.4 Delonix regia Hook (Bojer ex Hook) Delonix is a quick growing evergreen tree with slightly rough, grayish, brown bark, and a rather slender trunk, which usually soon divides into a number of 5
    • spreading, limb, bearing delicate feathery foliage 7-12 meter tall (Plate 1.4). It is planted in gardens, roadsides and at public places, as an ornamental shade tree. Plate 1.4: Delonix regia growing in Low Pollution Area with flowers in inset. Leaves - 15- 40 cm long, alternate, bipinnate compound, pinnae 8-20 pairs, leaflet 15-20 pairs. Flowers- 3 to 4 inches across petal, obviate, clawed, in terminal, simple or branched racemes, flowers red or orange in colour, the upper petal striped with yellow or white. Stamens- 10, exerted, red. Pods- 30-40 x 3 - 4.5 cm broadly linear, flat woody beaked, dark brown in colour. Seeds– Numerous, oblong, glabrous, smooth, white or creamy, mottled. Flowering- April-July. Fruiting - December.1.2.1.5 Peltophorum inerme Roxb. Peltophorum is evergreen tree, 8-20 meter tall, handsome, dark foliaged younger parts rusty brown or grayish tomentose, panicles of showy yellow flowers (Plate 1.5). It is usually planted in gardens and along the roadsides as an ornamental shade tree. Leaves –12-30 cm long, alternate, pinnae, 6-13 pairs. 6
    • Leaflets - 6-17 pairs, oblong, glabrous. Flowers- Bright yellow, in terminal racemose panicles. Plate 1.5: Blooming Peltophorum inerme growing in Low Pollution Area Stamens- 10, free, hairy at base golden-yellow. Pods- Lanceolate, 5-10 x 1.6 –2.2 cm, oblong, flat, hard, narrowed at both ends, indehiscent, woody, margin winged, rusty red in colour. Seeds– Usually 3-5, brown, obovate, oblong, compressed, smooth, flat and glabrous. Flowering- April-June. Fruiting– December- January.1.2.2 Flowering Time The data of intiating flowering for Caesalpinia pulcherrima, Cassia fistula, Cassia siamea, Delonix regia and Peltophorum inerme was noted for two consecutive years 2002 and 2003.1.2.3 Floral Morphology To study the floral morphology flowers were collected in between 9 to 11 AM from the height of 3 to 5 meters from the ground level. Hundred flowers were 7
    • collected from each plant species (25 flowers each from four different trees) from sampling sites in polythene bag sealed with adhesive tape and were brought to the laboratory. Measurement of length and breadth of sepals, petals, stamens and carpel were taken with a standard scale.1.2.4 Flower Colour The anthocyanin content of flowers, growing in different areas was determined following Drumm and Mohr (1978). For floral estimation 200 mg of petals were dipped into 5 cm3 of methanolic HCl (1%) v/v and kept overnight at 5 to 10 C (Stafford 1966). After centrifugation, the absorbance of supernatant was measured at 525 nm, in spectrophotometer. The anthocyanin content was expressed as absorbance per 100 mg fresh weight. Each mean value represent an average of three independent replicates.1.2.5 Floral Biomass Floral biomass was determined by collecting 100 flowers from each site from the height of 3 to 5 meters. Sampling was done in the morning hours between 9 to 10 am. Flowers were brought to the laboratory in polythene bags sealed with adhesive tape. After taking their fresh weight flowers placed in an oven at 80 C for 24 hours and later on the dry weight was recorded.1.2.6 Pollen Germination Freshly opened flowers were collected during 9 to 10 AM in polythene bags from Industrial Pollution Area (IPA), Vehicular Pollution Area (VPA) and Low Pollution Area (LPA) for pollen germination studies. Sucrose and boric acid solutions of different grades were prepared following Brewbakar and Kwack (1963). Pollen grains were placed in most suitable concentrations, i.e. (8% sucrose and 200 g of boric acid) on cavity slides, which were kept in petridish containing moist filter paper inside to maintain the appropriate relative humidity. The slides were observed under microscope at every one-hour interval to record the results. Pollen grains were considered germinated only, 8
    • when pollen tubes attained a size doubled of the grains. Ten random microscopic fields (10 x 10 X) in each of the slides were examined to determine the pollen germination. Pollen tube length and pollen diameter was measured using an Ocular Micrometer.1.2.7 Pollen Viability Pollen viability was determined by using 1 % TTC [2, 3, 5–Triphenyl- tetrazolium chloride] following Norton (1966). Pollen grains were incubated in 1 % TTC for 60 minutes at room temperature. The pollen grains were placed in a drop of this solution on a glass slides, with cover slip and these slides were kept in petriplates lined with moist filter paper and stored in a dark place. The numbers of pollen grains, which became reddish in colour, were recorded as viable.1.3 Results1.3.1 Flowering Time A delay in flowering time from 7 to 20 days was observed in all plants species in both IPA and VPA as compared to LPA. Maximum delayed flowering was noted in Cassia fistula. However, Caesalpina pulcherrima was found to be relatively unaffected (Table 1.1). Table 1.1: Delay in flowering period of tree species growing in different polluted areas of Indore city in comparison to Low Pollution Area Industrial Pollution Area Vehicular Pollution Area TPL* 539.15 g/ m3 TPL 506.81 g/ m3 Name of plant species Delay in Delay in Delay in Delay in Flowering Flowering Flowering Flowering 2003 2004 2003 2004 C. fistula 12-15 days 14-16 days 11-14 days 15-20 days C. siamea 15-20 days 16-18 days 10-12 days 8-10 days C. pulcherrima 7-10 days 5 -7 days 8-11 days 11-14 days D. regia 10-12 days 15-18 days 12-18 days 10-12 days P. inerme 8-10 days 9-11 days 15-18 days 18-21 days *TPL -Total Pollution Load (SO2 + NOx + SPM)1.3.2 Floral Morphology Flowers collected from polluted sites showed reduction in length and breadth of sepals and petals. Length of stamens and carpel was also noted reduced (Table 9
    • 1.2 to 1.6 and Fig. 1.1, 1.2 and 1.3). Maximum reduction was found in Peltophorum inerme, i.e. 26.71 and 59.0 %, 27.48 and 53.30 % in IPA and VPA respectively in length of sepals and petals. Whereas Caesalpinia pulcherrima showed minimum reduction, i.e. 3.27 % in sepals and 7.84, 13.27 % both in IPA and VPA. Table 1.2: Length and breadth (cm) of different floral parts of Cassia fistula LPA IPA VPA Parameters TPL* TPL* % Reduction TPL* % Reduction 308.02 g/ m3 539.15 g/ m3 506.81 g/ m3 Length of 1.22 1.14 1.18 sepals** 1.23 1.11 1.14 1.26 1.10 1.16 Average *** 1.23±0.16 1.11±0.16 9.75 % 1.16±0.16 5.69 % Breadth of 0.48 0.27 0.32 sepals** 0.49 0.30 0.36 0.47 0.32 0.38 Average*** 0.48±0.08 0.29±0.20 39.58 % 0.35±0.21 27.0 % Length of 2.83 1.18 1.87 petals** 2.86 1.14 1.74 2.84 2.20 1.69 Average*** 2.84±0.14 2.17±1.16 23.59 % 1.76±0.20 28.87 % Breadth of 1.08 0.52 0.92 petals** 1.49 0.60 0.69 1.08 0.94 0.84 Average*** 1.21±0.60 0.68±0.57 43.25 % 0.81±0.41 32.25 % **Length 2.15 1.11 1.98 of stamens 1.87 1.12 0.99 2.08 1.12 1.24 Average*** 2.03±0.46 0.90±0.53 44.82 % 1.40±0.74 30.87 % Length of 1.97 0.96 1.43 Carpel** 1.70 0.92 1.29 1.71 0.87 1.20 Average*** 1.79±0.35 0.91±0.55 49.16 % 1.30±0.02 27.37 %*TPL – Total Pollution Load, ** - Average of 100 flowers; *** - Average of 300 flowersLPA - Low Pollution Area, IPA- Industrial Pollution Area; VPA- Vehicular Pollution Area Table 1.3 : Length and breadth (cm) of different floral parts of Cassia siamea LPA IPA VPA Parameters TPL* TPL* % TPL* % 308.02 g/ m3 539.15 g/ m3 Reduction 506.81 g/ m3 Reduction Length of 1.30 1.10 1.12 sepals** 1.31 0.99 1.11 1.33 0.96 1.18 Average *** 1.31±0.14 1.01±0.20 22.90 % 1.13±0.57 13.74 % Breadth of 0.68 0.45 0.48 sepals** 0.67 0.46 0.47 0.69 0.47 0.49 Average*** 0.68±0.11 0.46±0.11 32.35 % 0.48±0.11 29.41 % Length of 2.43 0.97 0.99 petals** 2.40 0.98 1.21 2.41 1.12 1.10 Average*** 2.41±0.14 1.02±2.79 57.67 % 1.10±0.38 54.35 % 10
    • LPA IPA VPA Parameters TPL* TPL* % TPL* % 308.02 g/ m3 539.15 g/ m3 Reduction 506.81 g/ m3 Reduction Breadth of 1.20 0.60 0.69 petals** 1.21 0.58 0.61 1.19 0.59 0.59 Average*** 1.20±0.11 0.59±011 50.83 % 0.63±0.08 47.5 % **Length of 1.48 0.99 0.87 stamens 1.49 0.98 0.90 1.48 0.97 0.90 Average*** 1.48±0.08 0.98±0.11 33.78 % 0.90±0.08 39.18 % Length of 1.50 1.47 1.48 Carpel** 1.50 1.48 1.49 1.49 1.46 1.48 Average*** 1.50±0.08 1.47±0.11 2.0 % 1.48±0.08 1.33 %Table 1.4: Length and breadth (cm) of different floral parts of Caesalpinia pulcherrima LPA IPA VPA Parameters TPL* TPL* % Reduction TPL* % 308.02 g/ m3 539.15 g/ m3 506.81 g/ m3 Reduction Length of 1.22 1.18 1.19 sepals** 1.26 1.17 1.18 1.20 1.20 1.17 Average *** 1.22±0.20 1.18±0.16 3.27 % 1.18±0.11 3.27 % Breadth of 0.58 0.48 0.38 sepals** 0.49 0.40 0.47 0.48 0.46 0.48 Average*** 0.51±0.20 0.47±0.18 7.84 % 0.44±0.29 13.27 % Length of 2.84 2.81 1.90 petals** 2.85 2.80 1.99 2.82 2.82 2.00 Average*** 2.83±0.16 2.81±0.11 0.70 % 1.96±0.29 30.74 % Breadth of 0.64 0.59 0.48 petals** 0.59 0.58 0.61 0.62 0.60 0.49 Average*** 0.61±0.21 0.59±0.11 3.27 % 0.52±0.25 14.75 % **Length 4.6 3.9 4.00 of stamens 4.4 4.1 4.2 4.3 3.8 3.9 Average*** 4.4±0.16 3.9±0.37 11.36 % 4.0±0.37 8.33 % Length of 4.2 4.0 4.1 Carpel** 4.1 3.7 3.9 3.9 3.5 3.7 Average*** 4.0±0.57 3.7±0.46 7.5 % 3.9±0.51 2.5 % Table 1.5 : Length and breadth (cm) of different floral parts of Delonix regia LPA IPA VPA Parameters TPL* TPL* % Reduction TPL* % 308.02 g/ m3 539.15 g/ m3 506.81 g/ m3 Reduction Length of 2.30 2.00 2.13 sepals** 2.33 1.99 2.00 2.45 1.98 2.18 Average *** 2.36±0.34 1.99±0.11 15.67 % 2.10±0.37 11.01 % Breadth of 0.88 0.70 0.72 sepals** 0.80 0.71 0.71 0.69 0.69 0.72 Average*** 0.79±0.85 0.70±0.11 11.39 % 0.71±0.11 10.12 % 11
    • LPA IPA VPA Parameters TPL* TPL* % Reduction TPL* % 308.02 g/ m3 539.15 g/ m3 506.81 g/ m3 Reduction Length of 5.27 4.12 4.25 petals** 5.30 4.11 4.33 5.54 4.28 4.69 Average*** 5.37±0.47 4.17±0.66 22.34 % 4.42±0.59 17.69 % Breadth of 3.72 3.10 3.41 petals** 3.68 3.18 3.20 3.67 3.04 3.11 Average*** 3.69±0.20 3.10±0.30 15.98 % 3.57±0.21 3.25 % **Length 3.80 2.52 3.20 of stamens 3.70 3.60 2.99 3.90 2.80 3.40 Average*** 3.80±0.36 2.97±0.77 21.84 % 3.19±0.40 18.42 % Length of 4.30 4.70 4.40 Carpel** 3.90 2.87 3.80 4.80 3.80 4.40 Average*** 4.30±0.25 3.79±1.00 11.86 % 4.20±0.36 2.32 % Table 1.6 : Length and breadth (cm) of different floral parts of Peltophorum inerme LPA IPA VPA Parameters TPL* TPL* % Reduction TPL* % Reduction 308.02 g/ m3 539.15 g/ m3 506.81 g/ m3Length of 1.31 0.99 0.98sepals** 1.32 0.96 0.91 1.30 0.94 0.98Average *** 1.31±0.11 0.96±0.46 26.71 % 0.95±0.25 27.48 %Breadth of 0.70 0.37 0.30sepals** 0.69 0.30 0.36 0.67 0.32 0.25Average*** 0.68±0.14 0.33±0.20 51.47 % 0.30±0.27 55.88 %Length of 2.42 1.11 1.12petals** 2.41 0.99 1.11 2.43 0.96 1.18Average*** 2.42±0.11 0.99±0.34 59.0 % 1.13±0.23 53.30 %Breadth of 1.21 0.38 0.42petals** 1.19 0.45 0.47 1.42 0.46 0.50Average*** 1.27±0.40 0.43±0.21 66.14 % 0.46±0.24 63.77 %**Length of 1.40 0.86 0.89stamens 1.49 0.87 0.87 1.50 0.88 0.96Average*** 1.46±0.18 0.87±0.11 43.15 % 0.90±0.25 38.35 %Length of 1.70 0.84 0.85Carpel** 1.50 0.86 0.79 1.50 0.99 0.95Average*** 1.56±0.23 0.89±0.34 42.94 % 0.86±0.37 44.87 % The response of petals to air pollution was also similar to the sepals. The perusal of tables clearly indicates that there was more reduction in length of sepals and petals as compared to their width. The size of stamens and carpel was also affected by pollution stress. Out of five test species maximum reduction in length of stamens was noted in Cassia fistula, i.e. 44.82 and 30.87% in IPA and VPA respectively followed by Cassia siamea (Table 1.2 and 12
    • 1.3), whereas minimum reduction in stamen size was found in Caesalpiniapulcherrima, i.e. 11.36 and 8.33 % in IPA and VPA.Regarding carpel, maximum reduction was found in Cassia fistula, where thevalues were 49.16 and 27.37 % in IPA and VPA respectively. However theminimum reduction in carpel length was observed in Cassia siamea (Table 1.3).Overall it can be concluded that reduction in size of stamens and carpel wasmore significant than sepals and petals. Flowers growing in IPA found to bemore affected than that of roadside plants (Fig. 1.1, 1.2 and 1.3). Fig 1.1: % reduction in length and breadth of sepals over LPA 60 50 40 % Reduction 30 20 C.fistula C.siamea C.pulcherrima 10 D.regia P.inerme 0 length of sepals breadth of sepals IPA VPA Fig 1.2 : % reduction in length and breadth of petals over LPA. 70 60 50 Reduction 40 30 % C.fistula 20 C.siamea C.pulcherrima D.regia 10 P.inerme 0 IPAIPA VPA VPA Length of Petals Breadth of Petals Length of Petals Breadth of Petals Fig 1.3 : % reduction in length of stamens and carpel over LPA 50 C.fistula C.siamea 45 C.pulcherrima D.regia 40 P.inerme 35 30 % Reduction 25 20 15 10 5 0 IPALength of stamens IPA Length of carpel VPA Length of Stamens Length of Carpels Length of Stamens Length of Carpels 13
    • 1.3.3 Flower Colour The anthocyanins are known to impart colour to the flowers along with carotenoids. In present study, it is visualized that air pollutants affected the colour of flowers adversely. In general, an overall reducing trend was observed in the floral colour in all the five test species in both the polluted sites as compared to the reference area (Table 1.7 to 1.11). The data clearly indicates that the flowers developing in vehicular pollution areas were affected more adversely than the Industrially Polluted Area. It is inferred that there was an overall increment in pigment content in all five species with their increasing exposure time. However, when compared with the pigment content of reference area a reduction was noted. The maximum reduction in anthocyanin content in third day flower stage was observed in P. inerme in VPA, which was 27.45% followed by C. fistula (18.46%); while it was minimum in C. siamea, i.e. only 3.15%. When compared to differently polluted area the floral pigment to be more sensitive to vehicular pollution than industrial. This is true for all the five species. Thus it appears that air around road side is more toxic to flower than other areas in spite of low pollution load as compared to industrial area. Table 1.7 : Anthocyanin content (/mg fresh weight) of Cassia fistula LPA IPA VPA TPL*308.02 g/ m3 TPL* 539.15 g/ m3 TPL* 506.81 g/ m3 Time Increase Increase over Increase Increase over % Increase Increase over % exposure (in mg) 0 day (in mg) 0 day Reduction (in mg) 0 day Reduction 0 day 0.93 0.94 0.87 0.94 0.96 0.88 0.96 0.95 0.87 Average 0.95  0.16 0.00 0.95  0.11 0.00 0.00 0.87 0.08 0.00 8.42 1 day 0.96 0.98 0.94 0.95 0.98 0.96 0.97 0.97 0.95 Average 0.96 0.11 0.01 0.98  0.08 0.03 2.08 0.95 0.11 0.08 1.04 2ndday 1.24 1.04 1.06 1.22 1.06 1.06 1.23 1.05 1.09 Average 1.23 0.11 0.27 1.05  0.11 0.10 14.63 1.07 0.08 0.20 13.00 3rd day 1.31 1.20 1.05 1.29 1.21 1.08 1.30 1.20 1.07 Average 1.30  0.11 0.35 1.20 0.08 0.25 7.69 1.06  0.16 0.19 18.46 4th day 1.29 1.09 0.99 1.26 1.11 1.10 1.27 1.10 0.98 Average 1.27  0.14 0.28 1.10  0.11 0.15 13.38 1.05  0.34 0.13 21.25*TPL -Total Pollution Load, LPA - Low Pollution Area, IPA- Industrial Pollution Area; VPA- Vehicular Pollution Area 14
    • Table 1.8 : Anthocyanin content (/mg fresh weight) of Cassia siamea LPA IPA VPA TPL*308.02 g/ m3 TPL* 539.15 g/ m3 TPL* 506.81 g/ m3 Time Increase Increase over Increase Increase over % Increase Increase over %exposure (in mg) 0 day (in mg) 0 day Reduction (in mg) 0 day Reduction 0 day 0.89 0.74 0.68 0.88 0.75 0.68 0.89 0.74 0.67Average 0.89  0.08 0.00 0.74 0.08 0.00 16.85 0.67 0.08 0.00 23.59 1 day 0.89 0.94 0.89 0.91 0.93 0.70 0.90 0.95 0.70Average 0.90 0.11 0.01 0.94  0.11 0.20 4.4 0.70  0.08 0.02 22.222ndday 0.89 0.94 0.71 0.90 0.96 0.72 0.90 0.95 0.72Average 0.90  0.08 0.01 0.95  0.11 0.21 5.55 0.72 0.08 0.04 20.003rd day 0.94 0.96 0.98 0.96 0.95 0.98 0.95 0.97 0.97Average 0.95  0.11 0.06 0.96 0.11 0.22 1.05 0.98 0.08 0.03 3.154th day 1.06 1.04 1.07 1.06 1.06 1.09 1.09 1.03 1.08Average 1.07 0.12 0.12 1.05 0.14 0.3 1.86 1.08 0.11 0.4 0.93 Table 1.9 : Anthocyanin content (/mg fresh weight) of Caesalpinia pulcherrima LPA IPA VPA TPL*308.02 g/ m3 TPL* 539.15 g/ m3 TPL* 506.81 g/ m3 Time Increase Increase over Increase Increase over % Increase Increase over %exposure (in mg) 0 day (in mg) 0 day Reduction (in mg) 0 day Reduction 0 day 0.98 0.89 0.88 0.98 0.91 0.89 0.97 0.90 0.89Average 0.98 0.08 0.00 0.90 0.11 0.00 8.16 0.89 0.08 0.00 9.18 1 day 1.01 0.91 0.98 1.03 0.91 0.97 1.02 0.90 0.98Average 1.02  0.11 0.04 0.91  0.08 1.00 1.07 0.98 0.08 0.09 3.922ndday 1.03 1.03 0.98 1.04 1.02 0.99 1.04 1.01 0.99Average 1.04 0.08 0.06 1.02 0.11 12.0 1.92 0.99 0.08 0.10 4.803rd day 1.06 1.09 0.99 1.06 1.11 0.98 1.07 1.10 0.99Average 1.07  0.11 0.09 1.10 0.11 20.0 2.80 0.99  0.08 0.10 7.474th day 1.28 1.20 1.20 1.27 1.19 1.21 1.22 1.19 1.20Average 1.25  0.23 0.27 1.19  0.08 29.0 4.8 1.20 0.08 0.31 4.0 Table 1.10 : Anthocyanin content (/mg fresh weight) of Delonix regia LPA IPA VPA TPL*308.02 g/ m3 TPL* 539.15 g/ m3 TPL* 506.81 g/ m3 Time Increase Increase over Increase Increase over % Increase Increase over %exposure (in mg) 0 day (in mg) 0 day Reduction (in mg) 0 day Reduction 0 day 1.01 0.89 0.89 0.02 0.90 0.88 0.03 0.90 0.89Average 0.02 0.11 0.00 0.90 0.08 0.00 10.78 0.89 0.08 0.00 12.74 1 day 1.03 0.89 0.90 1.05 0.91 0.89 1.04 0.91 0.89Average 1.04 0.08 0.02 0.91 0.08 0.20 12.5 0.90 0.08 0.01 13.462ndday 1.20 1.19 0.90 1.22 1.20 1.06 1.21 1.21 1.07Average 1.21 0.08 0.17 1.20 0.11 0.17 0.82 1.08 0.08 0.18 11.53rd day 1.20 1.20 1.01 1.21 1.21 1.02 1.20 1.20 1.02Average 1.20 0.08 0.16 1.20 0.08 0.16 00 1.02 0.08 0.13 15.0 15
    • LPA IPA VPA TPL*308.02 g/ m3 TPL* 539.15 g/ m3 TPL* 506.81 g/ m3 Time Increase Increase over Increase Increase over % Increase Increase over % exposure (in mg) 0 day (in mg) 0 day Reduction (in mg) 0 day Reduction 4th day 1.09 1.00 1.00 1.10 1.01 1.01 1.11 1.00 1.00 Average 1.10 0.08 0.08 1.00 0.08 0.08 9.0 1.00 0.08 0.11 9.0 Table 1.11 : Anthocyanin content (/mg fresh weight) of Peltophorum inerme LPA IPA VPA TPL*308.02 g/ m3 TPL* 539.15 g/ m3 TPL* 506.81 g/ m3 Time Increase Increase over Increase Increase over % Increase Increase over % exposure (in mg) 0 day (in mg) 0 day Reduction (in mg) 0 day Reduction 0 day 0.78 0.74 0.67 0.78 0.73 0.68 0.79 0.73 0.68 Average 0.78 0.08 0.00 0.73 0.08 0.00 6.4 0.68 0.08 0.00 12.8 1 day 0.79 0.79 0.69 0.80 0.78 0.70 0.80 0.78 0.70 Average 0.80 0.08 0.02 0.78 0.08 0.05 2.5 0.70 0.08 0.02 12.5 2ndday 0.98 0.98 0.79 0.99 0.97 0.80 0.99 0.98 0.80 Average 0.99 0.21 0.21 0.980.08 0.25 1.0 0.80 0.08 0.10 19.19 3rd day 1.01 0.98 0.73 1.02 0.99 0.74 1.03 0.99 0.75 Average 1.02 0.14 0.24 0.99 0.08 0.26 2.94 0.74 0.11 0.06 27.45 4th day 0.99 0.98 0.74 0.98 0.97 0.76 0.99 0.97 0.75 Average 0.99  0.11 0.21 0.97 0.08 0.24 2.02 0.75 0.11 0.07 24.241.3.4 Floral Biomass Fresh and dry weights of flowers of different plant species are presented in Table 1.12 and % reduction is presented in Fig. 1.4. It is evident that maximum reduction in flower weight has taken place in vehicular area and minimum at industrial area. Out of five test species, it is observed that flowers of Delonix Fig 1.4 : % Reduction in fresh and dry weight of flower over LPA. 50 45 40 35 30 C.fistula % REDUCTION C.siamea 25 C.pulcherrima D.regia P.inerme 20 15 10 5 0 Fresh weight IPA IPA VPA Fresh Weight Dry Weight Fresh Weight Dry Weight 16
    • regia appeared to be more sensitive to air pollution, as regards the biomass, as there was 35.95 % and 43.77 % reduction in dry weight was noted respectively in both IPA and VPA. Minimum reduction was noted in Peltophorum inerme 19.40% and 27.53% in both IPA and VPA in dry weight as compared to unaffected area, i.e. LPA. These results once again proved the toxicity of the air pollutants (Fig. 1.4). Table 1.12 : Fresh and Dry weight (g) of 100 flowers Low Pollution Area Industrial Pollution Area Vehicular Pollution AreaName of plant TPL*308.02 g/m3 TPL* 539.15 g/m3 TPL* 506.81 g/m3 species Fresh wt. Dry Fresh wt. % Dry wt. % Fresh wt. % Dry wt. % wt. Red. Red. Red. Red.C. fistula 60.00 6.75 58.60 5.30 56.98 4.60 65.50 7.36 54.00 12.71 4.88 29.78 54.05 13.93 4.36 38.20 68.00 7.65 56.30 5.09 55.51 4.48 Average 64.50±3.26 7.25 ±0.37 56.30±1.75 5.09±0.52 55.51±1.39 4.48±1.37C. siamea 32.00 5.30 31.07 4.13 33.34 3.74 35.50 5.87 28.00 15.17 3.72 31.84 28.50 12.53 3.19 40.72 37.50 6.21 30.00 3.98 30.00 3.36 Average 35.00±2.00 5.79±0.81 29.69±1.49 3.94±0.54 30.61±1.90 3.43±0.61C. pulcherrima 25.00 3.19 24.61 2.62 24.82 2.50 28.50 3.63 28.00 6.59 2.98 22.09 22.00 20.91 2.21 37.56 31.00 3.95 26.30 2.79 20.00 2.01 Average 28.16±2.05 3.59±1.00 26.30±1.50 2.79±.43 22.27±3.79 2.24±0.20D. regia 200.0 43.72 198.01 29.43 197.82 27.53 215.0 46.99 195.05 5.18 28.99 35.95 175.00 11.68 24.35 43.77 211.0 46.12 196.53 29.21 180.00 25.05 Average 208.66±2.90 45.61±1.35 196.53±1.40 29.21±0.54 184.27±3.51 25.64±1.67P. inerme 35.00 11.64 33.46 11.11 33.92 10.42 40.50 13.46 30.00 19.32 9.96 19.40 28.00 21.24 8.60 27.20 42.50 14.13 31.73 10.53 31.00 9.52 Average 39.33±2.40 13.07±1.35 31.73±1.51 10.53±0.87 30.97±1.99 9.51±1.101.3.5 Pollen Germination Air pollution has also been found to affect pollen germination adversely in all test species (Table 1.13 and Fig. 1.5). The maximum reduction in percent germination of pollen grains was noted in Cassia siamea i.e. 31.51 in VPA, and minimum in Cistula fistula 19.15 % in IPA. Whereas maximum reduction was noted in C. pulcherrima 17.08% and minimum in Cassia siamea 0.97%. Thus different species respond differently two types of pollution.Table 1.13 : Pollen germination of studied plants growing in different polluted areas of Indore city LPA IPA VPA Name of plant TPL* 308.02 TPL* % Reduction TPL* % Reduction species g/m3 539.15 g/m3 506.81 g/m3C. fistula 74.33 2.03 60.09 1.93 19.15 % 63.00  4.66 15.24 %C. siamea 67.91 2.33 46.51  2.3 31.51 % 67.25 1.99 0.97 %C. pulcherrima 65.83 1.23 51.58 1.95 21.64 % 54.58 2.1 17.08 %D. regia 84.04 7.08 63.68 4.33 24.22 % 73.11 7.08 13.00 %P. inerme 84.98 1.34 62.72 3.46 26.19 % 73.28 5.38 13.76 % 17
    • 1.3.6 Pollen Size The size of pollen grains was found to be affected by pollution stress. (Table 1.14 and Fig.1.6). Higher percentage reduction was noted in IPA than in roadside plants. Maximum reduction in pollen size was observed in Delonix regia, i.e. 48.83 and 42.48 % respectively in IPA and VPA sites. While minimum reduction was noted in Peltophorum inerme 15.54 and 13.26 % respectively at IPA and VPA sites. It is evident from data presented in Table 1.6. that in IPA, air was more harmful for the pollens growth and development than VPA. A size wise reduction was also noted. The greater the size of the pollen, more was the reduction irrespective of the pollution area. Smaller size pollens were least affected like P. inerme. Table 1.14 : Pollen diameter () in plants growing in different polluted areas of Indore city LPA IPA VPA Name of plant TPL* 308.02 TPL* % Reduction TPL* % Reduction species g/m3 539.15 g/m3 506.81 g/m3C. fistula 53.1± 1.24 43.5 ± 1.25 18.07 % 45.2 ± 1.23 14.87 %C. siamea 57 ± 1.30 38.1 ± 1.39 33.15 % 38.8 ± 1.28 31.92 %C. pulcherrima 54 ± 1.20 37.1 ± 1.38 31.29 % 38.7 ± 1.27 28.33 %D. regia 64.5 ± 1.27 33.0 ± 1.27 48.83 % 37.1 ± 1.38 42.48 %P. inerme 52.1 ± 1.24 44 ± 1.25 15.54 % 45.0 ± 1.22 13.62 % Fig. 1.6 : % reduction in Pollen size over control. 60 50 40 % Reduction C.fistula C.siamea 30 C.pulcherrima D.regia P.inerme 20 10 0 IPA IPA VPA 18
    • 1.3.7 Pollen Tube Length Pollen tube length was found to be much lower in IPA as compared to VPA. A general reducing trend in pollen tube length in both polluted sites was recorded. (Table 1.15 and Fig. 1.7). The maximum reduction in pollen tube length was noted in Cassia siamea, i.e. 52.98, 46.51 % in IPA and VPA respectively. Whereas minimum reduction in IPA in Caesalpinia pulcherrima 19.17% and Delonix regia 16.16% in VPA respectively as compared to unaffected area. D. regia was least affected by pollution stress.Table 1.15 : Pollen tube length () of studied plants growing in different polluted areas of Indore city LPA IPA VPA Name of Plant TPL* 308.02 TPL* % Reduction TPL* % Reduction species g/m3 539.15 g/m3 506.81 g/m3C. fistula 176.1 ± 21.22 137.1 ± 19.08 22.20 % 138.5 ± 19.03 21.35 %C. siamea 218.0 ± 7.06 102.5 ± 8.23 52.98 % 116.6 ± 19.61 46.51 %C. pulcherrima 182.5 ± 30.41 147.5 ± 29.18 19.17 % 138.5 ± 15.80 31.67 %D. regia 177.5 ± 30.76 143.3 ± 12.11 19.26 % 148.0 ± 19.86 16.61 %P. inerme 182.6 ± 33.58 126.5 ± 22.76 30.72 % 138.5 ± 22.5 24.15 % Fig 1.7 : % reduction in pollen tube length over LPA. 60 50 40 % Reduction IPA 30 VPA 20 10 0 C.fistula C.siamea C.pulcherrima D.regia P.inerme1.3.8 Pollen Viability Pollen viability is a very important character to assess reproductive behaviour of plants. In present study it was noted to be reduced in both the polluted sites (Table 1.16 and Fig. 1.8). There was more reduction in pollen viability in VPA as compared to IPA. The maximum reduction in pollen viability was found in Peltophorum inerme 38.27 % and Caesalpinia pulcherrima, i.e. 38.29% in IPA and VPA and minimum reduction was recorded in Cassia siamea, i.e. 20.73 % 19
    • and 17.07 % in both IPA and VPA. Thus it appears that to urban air pollutants the least affected pollens grains were of Cassia siamea. Table 1.16 : Percent viable pollens of studied plants growing in different polluted areas of Indore city Low Pollution Area Industrial Pollution Area Vehicular Pollution Area TPL*308.02 g/ m3 TPL* 539.15 g/ m3 TPL* 506.81 g/ m3Name of Total no. Viable Non- Total no. Viable Non-viable % Red. in Total no. Viable Non-viable % Red. inplant species of pollens pollens viable of pollens pollens pollens viable of pollens pollens pollens viable pollens pollens pollensC. fistula 96 83 13 79 63 16 24.09 69 61 08 26.50C. siamea 90 82 08 74 65 09 20.73 79 68 11 17.07C. pulcherrima 98 94 04 76 67 09 28.72 64 58 06 38.29D. regia 84 73 11 70 55 15 31.50 70 57 13 21.91P. inerme 88 81 07 60 50 10 38.27 71 61 10 24.69 Fig 1.8 : % reduction in pollen viabilty over LPA. 60 50 C.fistula C.siamea C.pulcherrima 40 D.regia P.inerme % Reduction 30 20 10 IPA VPA 0 IPA 1.4 Discussion 1.4.1 Time of Flowering Air pollutants are influencing the plants in various ways. Apart from vegetative parts, reproductive parts are also showing significant variations under pollution stress. One of the most prominent features is delayed flowering in plants growing in polluted habitats. Pawar (1983) and Dubey (1985) have reported this in Mangferia indica, Delonix regia and Acacia arabica trees growing in industrially polluted area with predominance of SO2. Recently Chauhan et al. (2004) has also reported delayed flowering and reduced floral density in Cassia siamea growing along road side of Agra one of the highly polluted cities of our country. Thus the present findings are in confirmation with these earlier reports. Pawar and Dubey (1985) correlate this delay with air pollution stress because due to many physiological and bio-chemical alterations, less photosynthate is available for reproductive growth and development. 20
    • 1.4.2 Morphology of Flowers Higher value of Length and Breadth ratio (L/B) clearly indicated that there was more reduction in length of sepals and petals as compared to their width. Generally reduction in length of both floral parts, i.e. sepals and petals results has been noted maximum. The reduction in number of flower size and cone development due to the air pollution specially SO2 has been reported by many workers (Houston and Dochinger 1977, Beda 1982 and Ernst et al. 1985) Flower size reduction in calendula due to SO2 exposure has been reported (Singh et al. 1985 and Yunus et al. 1985). Joshi and Sikka (2002) have reported reduced fresh and dry weight in flowers of Cassia fistula, Delonix regia and Peltophorum inerme growing in differently polluted are of Indore city. Pollution induced changes in floral morphology of Cassia siamea has been reported recently by Chauhan et al. (2004). Higher reduction in size of stamens and carpel in comparison to sepals and petals can be attributed to their more complex physiological and biochemical requirements. Maximum flower size reduction in C. fistula as compared to other species is a result of its higher sensitivity to air pollution, which has been reported earlier on the basis of various morphological and phytochemical observations by many workers (Pawar 1982, Joshi 1989, Singh and Rao 1983, Agrawal 1986). Thus it is obvious that C. fistula is a very sensitive plant to urban air pollution not only regarding its vegetative and biochemical aspects but reproductive behaviour as well.1.4.3 Flower Colour Increment in floral pigment with their age can be attributed to the effect of light. Exposure of plants to white light increases the anthocyanin content in flowers resulted in their darkening (Stafford 1965 and Drumm and Mohr 1978). In the present study also the same pattern was observed. However the higher rates of reduction in anthocyanin pigment of flowers growing in polluted sites with their exposure time in comparison to low polluted area can be attributed to the phytotoxic activity of air pollutants. The decrease in floral colour in polluted 21
    • areas appears to be enzymatic in nature. Increased activities of glucosidase and poly-phenolonidase in the plant growing in mixed pollution area (Godzite 1967) and glycosidase (Bucher 1979) have been reported, which are known to reduce their pigments (Goodwin 1976). One of the reasons for plant wise variation in anthocyanin content reduction can be related to the thickness of petals. More reduction in P. inerme and C. fistula may be due their thin and delicates petals. Since the bright colour of flowers serve to attract the pollinators and thus ensure the pollination effectively. Fading of the flowers in such polluted habitats may also results in less fruiting and seed setting especially in entomophyllous flowers.1.4.4 Floral Biomass Decrease flower weight in polluted sites is related to reduction in floral size. Such changes in floral biomass can be either indirect effect of air pollution due to less allocation of photosynthates (Lechowicz 1987) or a direct effect of toxic gases on floral parts during their growth and development. There observations are in confirmation with earlier reports (Joshi and Sikka 2002). Higher sensitivity of Cassia fistula flower in comparison to rest of the species can be attributed to its overall sensitivity of plant, and can be accounted to the delicacy of floral parts, which remained totally exposed to pollutants to right from their initiation to full bloom in absence of leaves. Minimum alteration in C. siamea and P. inerme can be account on their resistant nature of their plants. These two plants have also been reported to have higher value of Air pollution Tolerance Index (Singh and Rao 1983 and Agrawal 1986).1.4.5 Pollen Characters The pollen grains are very sensitive to air pollution and thus have been used for monitoring of atmospheric pollution (Rosen 1983). The sensitivity of pollens to SO2 (Karmosky and Stairs 1974 and Varshney and Varshney 1981) and fluorides (Facteau and Rowe 1977) has been reported as poor germination and reduced tube growth. In present study also reduction in pollen grains size, viability, germination and tube length has been noted in all the plant species studied. These effects are 22
    • considered to be the influence of various air pollution combinations of IPA andVPA sites, effect of other pollutants cannot be denied too.Increased SO2 concentration is reported to reduce pollen germinationsignificantly (Dubey 1983 and Varshney and Varshney 1981, 1986). SimilarlyKrishnayya and Bedi (1986) have reported reduced pollen germination and seedviability in two species of Cassia growing near highways as a function of leadaccumulation thought they mainly consider it as an effect of lead. Reduction inpollen size, viability and shape reduction in pine pollens have been reported byFedatov et al. (1983). Reduced pollen viability in some vegetables as a functionof SO2 pollution in the vicinity of Mathura refinery was also reported byBhardwaj and Chauhan (1990). The present findings regarding the interaction ofpollen characteristics and urban pollutants are in confirmation with findings ofJoshi and Sikka (2002) and Chauhan et al. (2004). The highest reduction inpollen size in D. regia as compared to other fours species can be attributed tothe pollen size. Because D. regia pollens are bigger in their size and thus theyrequire more photosynthate to maintain it, which is poorly available underpollution stress. This might have resulted in higher reduction in size. Reductionin pollen size due to high pollution in Pinus sylvestris have been reported(Mamajev and Shkarlet 1972). Recently Chauhan et al. (2004) has also reportedchange in morphology especially in ornamentations of pollen grains of C.siamea pollen collected from high vehicular load. Such pollen grains failed toshow distinct colpi and reticulate sculpturing with comparison to less pollutedsites. They opined that this is due to the deposition of pollutants particularly,suspended particulates matter due to heavy movement of automobiles.But this interpretation does not seem to be logical because change in surfacecharacteristic might be a result of overall pollution load. Actually duringdevelopment stage; pollen grains are concealed in anther lobe. Hence these arenot coming in direct contact with particulate matter. Ornamentation and othermorphological features have been taken shape prior to anthesis. So thesechanges might have occurred before anthesis.In most of the studies carried out in areas exposed to industrially polluted, yet inmost instance it is accompanied by other pollutants and additive effects must bereckoned with Bonte (1982). Nakada et al. (1976) showed the in vitro studies 23
    • indicates that the addition of SO2 to NO2 or O3 or HCHO, considerably increasethe percentage inhibition compared with the action of each product examinedseparately.There is little information available on the mechanism of action of SO2 on thepollen tube. However Ma et al. (1973) have measured the pollen mitotic indexof Tradescantia paludosa treated in vitro by SO2 they assumed the SO2 brokedown the chromosomes of the pollen tube. Delayed and reduced floral yield ofcarnation and geranium species have been reported along with vegetativegrowth retardation (Feder 1970). Ozone induced inhibition in pollengermination and pollen tube growth has been observed (Feder 1981). Workdone by Mumford et al. (1972) suggests that O3 induces the autolysis ofstructural glycoproteins and stimulates amino-acid synthesis in pollen andinhibited germination by 40-90 %.Thus it can be concluded that the changes observed in present study in flowersand pollens grains are the results of cumulative effect of urban air pollutants, i.e.SO2, NOX and Photochemical oxidants along with particulates. 24
    • Air Pollution Impact on Fruits2.1 Introduction The union of the male and female generative cells, after pollination, to form the fertilized egg leads to the formation of fruits and seeds. A fruit is the mature female part specially ovary which may or may not include other parts of the flower. The seed is the ripened ovule contained within the fruit. Later on in due course of time germination of seeds give rise to new plants. Seeds are typically composed of three parts the embryo, endosperm and the seed coat. Fruits may contain one to several seeds. The term „fruit‟ and „seed‟ are often used loosely, for example the so called „seeds‟ of many members of poaceae are actually one seeded fruits. There are different types of fruits, depending on how they are derived. They may be fleshy, dry indehiscent, dehiscent, aggregate etc. The fruits of Leguminoceae are derived from single carpel with marginal placentation having one to many seeds. These fruits are dry dehiscent or indehiscent and commonly called Pods. Most angiospermic seeds have a seed coat derived from either two integuments or single integument of the ovule. In bitegmic seeds the term „testa‟ is applied only to the outer layer, formed from outer integument, the part formed from the inner integument being the tegment. Seed coat may be complex multilayered tissue or simply enlarged ovule wall. This generally includes a hard, protective layer formed from all or part of the testa. Corner (1976) has classified seed coat according to the position of this mechanical layer. In exotestal seedcoat the mechanical layer is formed from the outer epidermis of the outer integument and in endotegmic seed coat, it is derived from the inner wall of the inner integument. Some times the mechanical layer consists of one or more rows of elongated, palisade like cells, such as the macrosclerides in the exotesta of many leguminoceae, which is the family under study during the present research work. Apart from the obvious mechanical protective function, to prevent destruction of the seed by dehydration or predation, the seed coat often has important subsidiary functions, usually related to dispersal. These may bear corresponding 25
    • specialized structures. Like presence of wings in wind dispersal seeds and fleshy seeds for dispersal by animals. Of the many seeds produced by a plant, only a small proportion survives. Predation, rotting, falling in the wrong place or any of the many other natural and man made hazards besetting a seed. Those that do survive will sooner or later germinate. While the seed is dormant, all its processes are slowed down so as to utilize available limited food resources very economically to keep the embryo alive. When the dormancy is broken and the conditions are favorable for germination, the seed rapidly takes in water and the respiration rate rises back to normal as cell starts to grow and divide. The area of greatest growth at first is the root initial and young root soon pushes its way out through the seed coat. Such seeds are called as germinated. At germination the testa is ruptured and the radicle emerges through the micropyle. The seedling is the most Juvenile stage of the plant, immediately after germination, seedlings have a root (radicle) and a hypocotyls, which bears the cotyledons and plumules bud. This bud produces the stem and leaves, which soon resemble those of the mature plant. The cotyledons or seed leaves usually differ from the first foliage leaves. In large seeded dicotyledons such as the legumes the cotyledons are fleshy and swollen, with a food storage function. The overall physiology and biochemistry of sexual reproduction i.e. flowering, fruiting, seed setting and seed germination is influenced by various environmental constrains of which air pollution is one of the most significant factors. Looking to the deteriorating air quality the present study was planned to assess the impact of urban air pollution on fruits and seeds of the selected plant species.2.2 Experimental Apart from foliar injury plants also show changes in their reproductive parts too, in response to polluted air. This study was aimed to know the effects of air pollution on fruit morphology and seed quality. The colour, size and weight of fruits and seeds along with seed count and viability were studied. 26
    • 2.2.1 Colour of Pods Mature pods of C. fistula, C. siamea, C. pulcherrima, D. regia and P. inerme were collected during 2002, 2003and 2004 from the selected areas of Indore city from a height of 3 to 4 meter. Colour of pods and injury symptoms on them were recorded visually and compared with reference area.2.2.1 Size of Pods Pod size measurement was performed by taking 20 pods from five trees of each test species brought to the laboratory in polythene bags. Thus 100 pods from each species from every study area were collected. Length and Breadth of pods were recorded with the help of a standards measuring tape. In case of C. fistula in place of breadth diameter was measured.2.2.3. Weight of Pods Hundred pods each for year 2002, 2003 and 2004 were collected from different pollution areas along with Low pollution area were dried in oven at 80º C for 24 hours and their dry weight was recorded using an electronic balance. The results are presented as grams per pod.2.2.4 Seed Count The effect of airborne pollutants on seed per fruit of selected tree species was also studied. For this purpose seeds were taken out from the pods collected or dry weight measurement and seed number per pod was also recorded.2.2.5 Seed Viability Seed viability was tested following Cottrell (1947) to test the viability imbibed seed were cut, so that the embryo is bisected and then seed were placed in a 1.0% solution of 2,3,5 Triphenyl -2 H-tetrazolium chloride (TTC). Viable embryo releases hydrogen ion during respiration, which combines with TTC, imparting red or pink colour to seeds. The seeds in which embryo turned pink or red after 24 hours were considered as viable and their number were recorded. The test was conducted in petri plates containing filter paper. Four replicates of 27
    • 25 seeds per petri plates were used for the study. The results are presented as percent viability. Like other aerial parts of the plant fruit are also remain exposed to polluted air throughout their developmental span. This ranges from few months to years depending upon the nature of plants. During this prolonged exposure they interact with air pollutants resulting in the variation in various morphological features like shape, size and colour.2.3 Results2.3.1 Colour of Pods Like leaves the fruit also remain exposed to the ambient air during their developmental period, thus they too showed response to polluted air. The pods of all the test species collected from polluted sites appeared dark in colour as compared to the pods collected from low pollution area, which were less dark and shiny. It was also observed that the colour of the pods growing in Industrial Pollution area affected more than pods collected from Vehicular Pollution Area. In most of pods their normal dark brown colour has turned in to dark brown to black due to the interaction of pollutants and deposition of particulate matter on them. Chlorotic and necrotic spots with tip burn were also observed in some pods of C. pulcherrima in the polluted areas (Table 2.1, Plate-2.1 to 2.5). Table 2.1: Colour of pods collected from different polluted areas of Indore city Name of Plant Low Pollution Area Industrial Pollution Vehicular Pollution species TPL* 308.02 g/m3 Area Area TPL* 539.15 g/ m3 TPL* 506.81 g/ m3 C. fistula Blackish-brown Dark black brown Dark black brown C. siamea Brownish Light brown Light brown C. pulcherrima Brown Dark brown Dark brown D. regia Dark black Dark black brown Dark black brown P. inerme Shiny blackish-brown Blackish brown Blackish brown *TPL -Total Pollution Load 28
    • Plate-2.1: Cassia fistula pods showing Plate-2.2: Cassia siamea pods showing colour change and size reduction. colour change and size reduction. Plate-2.3: Caesalpinia pulcherrima pods Plate-2.4: Peltoforum inerme pods showing colour change and size showing colour change and size reduction. reduction. Plate-2.5: Delonix regia pods showing size reduction.2.3.2. Size of Pods The polluted air has affected the size of the pods. There was a reduction in length as well as breadth of the pods in plants growing in different polluted sites. It is evident from the data presented in Table 2.2 and Fig. 2.1 that the reduction in pod length was more in Industrial Polluted Area as compared to Vehicular Polluted Area. 29
    • Table 2.2 : Length and Breadth Ratio of pods collected from different polluted areas of Indore city Low Pollution Area Industrial Pollution Area Vehicular Pollution AreaName of Y TPL*308.02 g/m3 TPL* 539.15 g/m3 TPL* 506.81 g/m3Plant Especies A L B L/B L B L/B % % Red L B L/B % % Red R Ratio Ratio Red (B) Ratio Red (B) (L) (L)C. S1 41.00 8.00 5.12 34.85 7.34 4.74 37.87 7.87 4.81fistula S2 40.32 7.99 5.04 38.92 7.54 5.16 36.45 7.33 4.71 S3 41.24 8.09 5.09 37.99 7.42 5.11 39.20 8.13 4.82 A 40.85 8.02 5.09 37.25 7.43 5.01 8.81 7.35 37.84 7.91 4.78 7.36 1.37 ±0.24 ±0.10 ±0.20 0.20 ±1.10 ±0.48 ±0.52 ±0.68 ±0.49C. S1 17.42 1.20 14.51 15.24 1.1 13.85 16.27 1.20 13.5siamea S2 17.36 1.30 13.35 14.74 0.9 16.37 16.98 1.10 15.4 S3 17.28 1.20 14.40 16.40 0.8 20.50 16.54 1.10 15.0 A 17.35 1.20 14.08 15.46 0.9 16.90 10.89 18.18 16.59 1.10 14.67 4.38 8.33 ±0.10 ±0.01 ±0.08 ±1.48 ±0.01 ±3.25 ±0.59 ±0.01 ±0.67C. S1 10.1 1.5 6.73 9.54 1.1 6.81 9.62 1.40 6.87pulche- S2 9.74 1.4 6.95 8.99 1.3 6.91 9.70 1.32 7.34rrima S3 10.3 1.6 6.27 9.39 1.5 6.26 9.02 1.20 7.51 A 10.04 1.5 6.65 9.30 1.4 6.66 7.37 6.66 9.44 1.30 7.2 5.97 13.3 ±1.29 ±1.11 ±1.35 ±1.29 ±1.10 ±1.54 ±1.78 ±1.15 ±1.29D. S1 39.25 3.15 12.46 37.46 3.10 12.08 38.24 3.12 12.25regia S2 38.00 3.50 10.85 37.89 3.48 10.88 38.43 3.34 11.50 S3 37.75 3.60 10.48 38.20 3.56 10.73 37.09 3.52 10.70 A 38.33 3.48 11.05 37.85 3.43 11.06 1.25 1.45 37.42 3.31 11.5 1.06 4.88 ±0.83 ±0.41 ±0.56 ±1.20 ±1.38 ±1.06 ±0.42 ±0.08 ±1.08P. S1 9.60 2.2 4.36 7.43 1.7 4.36 8.29 1.9 4.36inerme S2 9.77 2.0 4.88 8.24 2.00 4.12 8.74 2.1 4.16 S3 9.82 2.1 4.67 9.47 1.9 4.98 9.59 1.9 5.04 A 9.73 2.1 4.64 8.38 1.85 4.60 14.40 11.90 8.87 1.9 4.44 8.83 9.52 ±0.43 ±0.02 ±0.58 ±1.28 0.42 0.28 ±0.93 0.20 0.43 *TPL -Total Pollution Load, L – Length, B – Breadth, Red – Reduction, Sampling year - S1 –2002, S2 –2003, S3–2004 ; A – Average Values Maximum reduction in pod length was noted in P. inerme where it was 14.40 % and 8.83% respectively in IPA and VPA with reference to LPA. Whereas minimum reduction was recorded in D. regia where the values were 1.25 % and 1.06% respectively in IPA and VPA. The rest of two species of Cassia appeared more or less affected similarly at both sites. The breadths of the pods were also found decreased in all the species. Maximum reduction in breadth of the pod was recorded in C. siamea (18.18%) and C. pulcherrima (13.30%) respectively in IPA and VPA. Regarding breadth of pod C. fistula found to be affected least in VPA. 30
    • The L/B ratio of pods was also changed (Table 2.2). There was a slight increase in the ratio for C. siamea, C. pulcherrima and D. regia. However this ratio decreased in P. inerme and C. fistula showed that the pods breadth was comparatively more affected than length in most of the species studied. Thus it can be concluded that there was overall growth retardation in pods of all the test species growing in polluted areas.2.3.3 Weight of Pods Dry weight of pods is presented in Table 2.3. It can be seen from the table that dry weight of pods has also been reduced in all the plant species. The maximum reduction was observed in C. siamea i.e. 44.0 % in IPA and 30.30 % in VPA and minimum in C. pulcherrima, i.e 8.96% in IPA and 8.79% in VPA respectively. Whereas D. regia and P. inerme showed more than 20 % reduction in dry weight. Areas wise there was more reduction in pod dry weight in Industrial area than Vehicular area (Fig. 2.3). Table 2.3 : Dry weight of pods collected from different polluted areas of Indore city Name of LPA IPA VPA Plant Year TPL* 308.02 TPL* 539.15 % Reduction TPL* 506.81 % Reduction species g/m3 g/m3 g/m3 C. fistula 2002 67.92 51.78 50.88 2003 65.70 54.48 53.74 2004 63.82 52.72 55.34 Avg. 65.81±2.80 52.99 ± 1.25 19.48 % 53.32 ± 3.40 18.97 % C. pulcherrima 2002 12.37 11.98 9.09 2003 11.69 10.23 10.87 2004 10.42 9.17 11.48 Avg. 11.49±0.65 10.46 ± 1.34 8.96 % 10.48 ± 1.02 8.97% C. siamea 2002 12.97 7.28 8.98 2003 13.11 7.39 9.59 2004 13.53 7.47 9.04 Avg. 13.20±0.05 7.38±0.006 44.0 % 9.20 ±0.05 30.30% D. regia 2002 77.93 51.27 57.64 2003 68.43 54.25 51.37 2004 67.71 56.79 58.45 Avg. 71.35±2.95 54.10±5.08 24.17% 55.82  2.95 21.76% P. inerme 2002 10.37 8.52 8.63 2003 10.87 6.56 7.48 2004 9.24 7.88 7.69 Avg. 10.16±0.09 7.65±0.66 24.70% 7.93  0.15 21.91%* TPL - Total Pollution Load, ** Avg. - Average of 100 pods 31
    • Fig. 2.3 : % reduction in dry weight of pods over LPA. 50 C.fistula 45 C.siamea C.pulcherrima D.regia 40 P.inerme 35 30 Reduction 25 % 20 15 10 5 0 IPA VPA2.3.4. Seed count A perusal of Table 2.4 to 2.8 indicates that there was a reduction in seed number, which ranges from 20.55 % to 3.15 %. The maximum lowering in seed per pod was noted in C. siamea in Industrial area, while the minimum reduction was recorded for C. pulcherrima in VPA. The response of C. fistula, D. regia and P. inerme was almost same in both the polluted sites (Fig. 2.4). Fig. 2.4 : % reduction in Seed/pod over LPA. 25 C.fistula C.siamea C.pulcherrima D.regia 20 P.inerme 15 % Reduction 10 5 0 IPA VPA Further it is also evident that number of unhealthy seeds per pod is high in both the polluted sites in comparison with reference area. It clearly indicates that whatever be the nature of the pollutant it adversely influenced the seed number and quality. On the overall basis it can be stated that the colour, size, shape, weight and number of seeds per pod all were adversely affected by air pollution prevailing 32
    • in the area. Higher reduction in industrial pollution area in above parameters in comparison to vehicular pollution area corresponds to the preventing pollution. Table 2.4 : Healthy and unhealthy seeds per pod in Cassia fistula growing in different polluted areas of Indore city Low Pollution Area Industrial Pollution Area Vehicular Pollution Area TPL* 308.08 g/m3 TPL* 539.15 g/m3 TPL* 506.81 g/m3 No. of No. of No. of No. of No. of No. of No. of No. of No. of seed per healthy unhealthy seed per healthy unhealthy seed per healthy unhealthy Year pod seed seed pod seed seed pod seed seed 2002 46.44 46.00 0.44 46.00 44.87 1.13 45.80 44.68 1.12 2003 46.96 46.28 0.68 45.98 44.88 1.10 45.00 43.96 1.04 2004 62.09 51.51 0.68 45.04 43.89 1.15 46.12 45.03 1.09 51.83 47.93 0.60 45.67 44.54 1.12 45.64 44.55 1.08 3.69 2.18 0.46 0.92 0.93 0.20 .92 0.89 0.65 #11.88 % #11.94% *TPL -Total Pollution Load, # % Reduction Table 2.5 : Healthy and unhealthy seeds per pod in Cassia siamea growing in different polluted areas of Indore city Low Pollution Area Industrial Pollution Area Vehicular Pollution Area TPL* 308.08 g/m3 TPL* 539.15 g/m3 TPL* 506.81 g/m3 No. of No. of No. of No. of No. of No. of No. of No. of No. ofYear seed healthy unhealthy seed healthy unhealthy seed per healthy unhealthy per pod seed seed per pod seed seed pod seed seed 2002 23.88 22.64 0.16 17.20 16.96 0.20 19.48 18.52 19.38 2003 20.12 19.88 0.16 17.08 17.00 0.08 22.36 21.84 0.28 2004 21.96 21.72 0.24 17.06 17.04 0.20 20.92 20.02 0.72 21.65 21.41 0.18 17.20 17.12 0.16 20.92 20.18 6.79  1.64  1.43  0.17  0.41 0.41 0.32 1.38 1.52 2.94 #5.08% #3.37% Table 2.6 : Healthy and unhealthy seeds per pod in Caesalpinia pulcherrima growing in different polluted areas of Indore city Low Pollution Area Industrial Pollution Area Vehicular Pollution Area TPL* 308.08 g/m3 TPL* 539.15 g/m3 TPL* 506.81 g/m3 No. of No. of No. of No. of No. of No. of No. of No. of No. of seed healthy unhealthy seed healthy unhealthy seed healthy unhealthyYear per pod seed seed per pod seed seed per pod seed seed2002 8.20 8.12 0.12 7.96 7.44 0.52 7.90 7.34 0.562003 8.28 8.16 0.12 8.00 7.60 0.56 7.99 7.51 0.482004 824 8.16 0.08 7.80 7.32 0.48 8.05 7.46 0.59 8.24 8.14 0.10 7.92 7.45 0.52 7.98 7.43 0.54  0.23  0.20  0.20  0.49  0.49 0.23  0.32  0.28  0.29 #3.88% #3.15% Table 2.7 : Healthy and unhealthy seeds per pod in Delonix regia growing in different polluted areas of Indore city Low Pollution Area Industrial Pollution Area Vehicular Pollution Area TPL* 308.08 g/m3 TPL* 539.15 g/m3 TPL* 506.81 g/m3 No. of No. of No. of No. of No. of No. of No. of No. of No. ofYear seed healthy unhealthy seed healthy unhealthy seed healthy unhealthy per pod seed seed per pod seed seed per pod seed seed2002 24.40 24.28 0.12 20.24 20.00 0.24 21.24 20.80 0.362003 23.28 22.96 0.32 20.36 20.04 0.32 23.08 22.28 0.082004 23.20 22.36 0.34 20.44 20.02 0.24 22.25 21.75 0.45 23.96 22.84 0.42 20.34 20.02 0.26 22.28 21.94 0.29  0.48  0.84  0.46  0.38  0.16  0.25  0.43  0.53  1.15 #15.10% #7.01% 33
    • Table 2.8 : Healthy and unhealthy seeds per pod in Peltophorum inerme growing in different polluted areas of Indore city Low Pollution Area Industrial Pollution Area Vehicular Pollution Area TPL* 308.08 g/m3 TPL* 539.15 g/m3 TPL* 506.81 g/m3 No. of No. of No. of No. of No. of No. of No. of No. of No. of seed healthy unhealthy seed per healthy unhealthy seed healthy unhealthyYear per pod seed seed pod seed seed per pod seed seed2002 2.24 2.05 0.19 2.12 1.80 0.32 2.18 1.90 0.282003 2.48 2.28 0.20 2.22 1.84 0.38 2.23 1.89 0.342004 2.01 2.17 0.24 2.27 1.83 0.44 2.29 1.81 0.48 2.57 2.36 0.21 2.20 1.82 0.38 2.23 1.86 0.36  0.80  0.60  0.20  0.33  0.18  0.28  0.27  0.25  0.17 #14.39% #13.22%*TPL -Total Pollution Load, # % Reduction2.3.5 Seed Viability Perusal of Table 2.9 and Fig. 2.5 revealed that air pollution has adversely affected seed viability. The seed viability was more reduced in Industrial Polluted Area than Vehicular Pollution Area. Maximum reduction in seed viability was observed in D. regia i.e. 8.04% in IPA, and minimum reduction in seed viability was recorded in C. pulcherrima, i.e. 2.22% in IPA. While in VPA maximum reduction in seed viability was noted in C. siamea, i.e. 6.3 and minimum in C. fistula 3.29%.2.4 Discussion Significant morphological and physiological changes in the leaves exposed to air pollutants have been extensively worked out by many workers (Jacbson and Hill 1970, Chaphekar 1972, Sharma 1976, Pawar and Dubey 1983, Pandya and Bedi 1986 and Rangrajan et al. 1979). These alterations are not restricted to vegetative parts only but greatly influence reproductive structures too. Changes in pod colour, size and dry weight were noticed during the present study this has also affected the overall fruit and seed quality. The darkening of the pods observed in plants facing air pollution may be due to the impact of various gaseous as well as particulates present in the air. Pods of plants such as D. regia and C. fistula remain exposed to air for months together. Thus interacting with pollutants for longer duration, which result in alteration in the physiological processes of fruit ripening, which may cause deviation in the normal colour of the fruit. The darkening of the colour of pods may be 34
    • attributed to the loss of photosynthetic pigments as a result of harmful effects of pollutants during their developmental stage and deposition of chemically active particulates on them. Table 2.9 : Seed viability of plant species collected from different polluted areas of Indore city LPA IPA VPA Name of TPL* 308.08 g/m3 TPL* 539.15 g/m3 TPL* 506.81 g/m3 plant Year % of viable seeds# % of viable % Reduction % of viable % Reduction species seeds# seeds# C. fistula 2002 92 80 84 2003 88 84 88 2004 90 88 92 Avg. 91.01.82 842.30 7.69 % 882.16 3.29 % C. siamea 2002 96 92 88 2003 92 88 92 2004 94 90 84 Avg. 941.63 901.63 4.25% 882.30 6.38% C. pulcherrima 2002 93 88 82 2003 89 92 92 2004 88 94 84 Avg. 902.30 883.05 2.22% 862.44 4.44% D. regia 2002 92 84 80 2003 87 76 84 2004 84 80 80 Avg. 872.30 802.30 8.04% 842.30 3.44% P. inerme 2002 96 91 92 2003 92 89 88 2004 88 87 80 Avg. 922.30 891.63 3.33% 860.26 8.69%*TPL - Total Pollution Load, #- Average of 300 seeds Reduction in fruit size, seed number and dry weight as a function of different pollutants has been reported earlier (Houston and Dochinger 1977, Murdy 1979, Murdy and Ragsdale 1980). These findings are in confirmation with present study. Further Claussen (1970) and Cluster (1982) held responsible automobile exhaust with NO2 and CO for such changes. Murdy and Ragsdale (1980) have also shown that in Gernanium SO2 damages sexual reproduction in terms of decreased seed set. SO2 induced fertility changes in Lepidium virginicum have been reported (Murdy 1979). 35
    • Khan and Khan (1991) working on the impact of air pollution has also observedreduced fruiting, smaller size and low weight of Tomato fruits. Reduction infruit per plant, seed out put and seed per fruit in Brassica juncia growing near athermal power plant have been reported by Saquib and Khan (1999). All thesereports support present findings.Plants growing in Industrially polluted air with predominance of SO2 showedreduced flowering and fruiting up to the extent of total absence of flowers andfruits in some cases has also been reported earlier (Pawar 1982). Khan andKhan (1991) and Saquib and Khan (1999) concentrated their studies onvegetables only. However, no such work has been conducted with tree speciesexcept (Rao 1972, Pawar 1982, Pawar and Dubey 1983 and Sikka and Joshi2002). The result of the present study also reveals a reduction in fruit size, andseed per pod, which agrees with earlier observations.Recently Chauhan et al. (2004) has reported at Agra reduction in fruit length,fruit number and seed per fruit in Cassia siamea and accounted it to automobilepollution. More damage in fruits and seeds in industrially polluted area ascompared to Vehicular pollution area can be attributed to the total higherpollution load in the ambient air of preceding area.Reduction in pod number and mean pod weight in Cicer arietinum exposed to80 ppb ozone has been reported by Singh and Rao (1982). Since the urban aircontains a mixture of gaseous and particulate pollutants. The effects observedon fruits and seed morphology in present study are cumulative in which the roleof ozone cannot be ignored, which has became a common air pollutant of urbanareas.In total the reducing trend in fruit size, seed quality and seed number is a resultof cumulative effect of various air pollutants present in ambient air of Indorecity of which many of them can not be monitored due to the lack of facilities. 36
    • Air Pollution Impact on Seed Quality and Germination3.1 Introduction Ever since the origin of a species, the inherent capacity of reproduction has resulted in its spread thus making the distribution a dynamic process (Ridley 1930). When a plant has reached a certain stage of development, its growing point may under certain conditions, changes from the vegetative to reproductive phase. The success of a species in propagating itself by seeds depends primarily on the number and viability of the seeds produced by parents, provided that those seeds find conditions suitable for germination and subsequent growth. Morphologically seed is a mature integumented megasporangium. The size and shape of the seed depends on the form of ovary, conditions under which the parent plant grows during seed formation and obviously, on the species. Other factors, which determine the size and shape of seeds, are the size of the embryo, the amount of endosperm present and to what extent other tissues participate in the seed structure (Mayer and Mayber 1963). The seed according to Berlyn (1972), is a packet of energy, some of which is in the form of information, it is the state of minimum entropy in the life cycle of angiosperm and gymnosperms. A viable seed is one, which can germinate under favorable conditions provided that any dormancy if present may be removed (Roberts 1972). The state of dormancy is overtly terminated when active metabolism, synthesis and finally growth are resumed. In seeds the resumption of these activities is identified with the initiation of various metabolic activities leading to germination. Such post germinative activities can only take place in environments within which the parent plant could function properly. One of the requirements for germination is an adequate moisture supply yet which would not interfere with the gaseous exchange, which is essential for aerobic respiration and adequate supply of metabolic energy. Another such requirement is for “normal” temperature, i.e. within the range, which is suitable for growth of more mature seedling. Ostensibly, therefore exposure of seeds to 37
    • environment consisting of adequate moisture, aeration and „normal‟ temperature, should suffice for germination to take place. Angiospermic seed may appear simple externally but possesses a complex eco- physiology for its further resumption of growth, primarily its germination. Most of the authors called seed germination as “the sprouting of seeds, or resumption of growth by dormant embryo”. It is that group of processes which cause the sudden transformation of dry seed in to the young seedling (Mayer 1953). Thus germination is the period during which physiological processes are initiated in the seed, leading to the elongation of cells and the formation of new cells, tissues and organs, i.e. the period between hydration and the onset of meristematic activities and finally differentiation of cell and organogensis. Hence, the germination of seed may be defined as the sequential series of morphogenetic events that result in the transformation of an embryo into seedling. It is a half closed system i.e. initiated when quiescent embryo is reactivated, but the terminal end of the system is open because the point where germination ends and seedling growth commences is undefined (Mayer and Mayber 1963). Seeds with special germination requirements are said to be dormant or blocked (Toole 1961). It has to be recognized that internal conditions of the seed may also be considered as important factor in determining its germination (Stiles and cocking 1961). According to Amen (1966), seed dormancy is an adaptive mechanism of growth cessation. Since the environmental conditions before and during seed development are very important. The present study was conducted to know the effects of air pollution on morphological and physiological aspects of seed and their germination.3.2 Experimental In order to study the effects of urban air pollution on the seed quality, quantity and physiology, the following parameters were studied: 38
    • 3.2.1 Seed Colour Seed from the pods of C. fistula, C. siamea, C. pulcherrima, D. regia and P. inerme were collected from different study areas of Indore city and seed colour was observed and recorded with reference to low pollution area.3.2.2 Seed Weight For seed weight measurement seeds from the pods of test plants collected from various sampling stations were taken out in distinct lots from each lot 100 seeds were drawn randomly in three replicates and weighed in grams. The seed lot with high seed weight is considered as Vigorous.3.2.3 Seed Density Seed density is a very important parameter to know their quality. To determine the density of the seeds, a definite quantity of kerosene oil is poured in a measuring cylinder and then initial level was noted. There after pre weighed seed sample was added to the measuring cylinder and the rise in level of the kerosene was noted. The seed density was calculated as Weight of the seeds Seed density = Volume of the seeds3.2.4 Seed Soundness This is another test to know about the vigor of the seed. If the number of shriveled, under-sized, under-developed, discolored and insect damaged seeds are more in a seed lot it is considered poor. To study the soundness of seeds collected from various polluted sites the above-mentioned characters were observed in lots of 100 seed in triplicates.3.2.5 Seed Germination Since the seed of the entire five test plants are known to possess physical dormancy (Cassia fistula, Athayia 1990 and Todaria and Negi 1992, Cassia siamea, Todaria and Negi 1992, Delonix regia, Gill et al. 1981, Peltophorum inerme, Anoliefo and Gill 1992, Caesalpina pulcherrima, Jones and Geneve 39
    • 1995). They were subjected to mechanical scarification by rubbing them on a sand paper no.10 before placing them for germination test. The germination test was conducted at National Soybean Research Center (NRCS), Khandwa Road, Indore. For the test germination paper was soaked in distilled water than 50 seeds in three replicates each were placed on the paper. Thiarm is used in small quantity to protect seeds from fungal contamination during germination. All seed lots were placed in a seed germinator. The seed were grown in four replicates of 50 seeds. Numbers of germinated seed were counted daily.3.3. RESULTS3.3.1 Seed Colour Like pods, which were exposed to air pollution, the seed colour was also found to be changed. In both the polluted areas the seed colour became dark, were as seeds of reference area were light in colour and shiny (Table 3.1). This was true for all five species. The maximum colour darkening was noted in C. fistula, while minimum colour change was recorded in D. regia and P. inerme as compared to seeds collected from Low pollution area. Table 3.1: Colour of seeds collected from different polluted areas of Indore city Name of plant Low Pollution Area Industrial Pollution Vehicular Pollution species TPL* Area Area 308.02 g/ m3 TPL* TPL* 539.15 g/ m3 506.81 g/ m3 C. fistula Light brown and Dark blackish- brown Dark blackish- brown shiny C. siamea Blackish-brown and Blackish-brown Blackish-brown shiny C. pulcherrima Greenish-brown Dark greenish and Dark greenish and blackish-brown blackish- brown D. regia Ivory-greenish Dark greenish Dark greenish P. inerme Light brown Light brown and Light brown and some darker some darker *TPL -Total Pollution Load 40
    • Table 3.2: Weight of 100 seeds (g) collected from different polluted areas of Indore city LPA IPA VPA Name of plant TPL* 308.02 TPL* % Reduction TPL* % Reduction species g/m3 539.15 g/m3 506.81 g/m3C. fistula 17.080 16.00 12.125 17.110 15.125 13.080 17.150 15.075 13.110 Avg. 17.113  0.025 15.40 0.89 10.00 12.771 .925 25.37C. siamea 4.500 4.180 4.100 4.350 4.200 4.080 4.600 4.250 4.115 Avg. 15.451 0.619 13.526  1.034 12.45 13.203 0.98 14.54C. pulcherrima 15.739 13.150 13.286 15.200 13.100 12.600 15.415 14.330 13.725 Avg. 15.451 0.619 13.526  1.034 12.45 13.203  0.98 14.54D. regia 36.670 31.680 33.320 35.500 32.320 32.870 36.200 31.983 32.995 Avg. 36.123 0.911 31.994  0.647 11.43 33.061  0.505 8.47P. inerme 5.105 4.500 4.100 5.250 4.420 4.050 5.360 4.330 4.085 Avg. 5.238  0.421 4.416  0.036 15.69 4.078  0.194 22.14  TPL – Total Pollution Load 41
    • Table 3.3 : Density of seeds collected from different polluted areas of Indore cityName of Plant LPA IPA % Reduction VPA % species TPL* 308.02 TPL* TPL* Reduction g/m3 539.15 g/m3 506.81 g/m3C. fistula 1.73 1.47 1.60 1.70 1.40 1 .65 1.65 1.35 1.60 Avg. 1.69 0.08 1.40 0.33 17.15 1.61 0.20 4.73C. siamea 1.09 1.08 1.08 1.05 1.08 1.00 1.09 1.00 1.08 Avg. 1.07 0.20 1.05 0.27 21.83 1.05 0.27 1.86C. pulcherrima 1.32 1.12 1.11 1.30 1.10 1.10 1.25 1.13 1.15 Avg. 1.29 0.23 1.11 0.01 13.95 1.12 0.20 13.17D. regia 1.43 1.13 1.15 1.400 1.10 1.10 1.45 1.12 1.12 Avg. 1.42 0.20 1.11 0.16 21.83 1.12  0.18 21.12P. inerme 1.31 1.05 1.20 1.20 1.00 1.30 1.28 1.05 18.25 1.25 Avg. 1.26 0.29 1.03 0.21 1.25 0.25 0.793  TPL – Total Pollution Load Table 3.4: Soundness of seeds collected from different polluted areas of Indore cityName of Plant Low Pollution Area Industrial Pollution Area Vehicular Pollution Area species TPL* 308.02 g/m3 TPL* 539.15 g/m3 TPL* 506.81 g/m3 No. of No. of un- No. of % No. of un- No. of % No. of un- healthy healthy healthy Decrease healthy healthy Decrease healthy seeds** seeds seeds seeds seeds seedsC. fistula 92 08 82 10.86 18 86 6.52 14C. siamea 93 07 78 16.12 22 82 11.82 18C. pulcherrima 93 07 88 4.16 12 84 9.67 16D. regia 94 06 88 6.38 12 91 3.19 09P. inerme 96 04 90 6.25 10 93 3.12 07*TPL – Total Pollution Load** Total number of unhealthy seeds = (a + b + c + d + e) Where, Shriveled seeds (a), Under-sized seeds (b), Under-developed seeds (c), Discoloured seeds (d) and Insect damaged seeds (e). 42
    • Table 3.5: % Reduction in seed germination in different polluted areas of Indore city Low Pollution Industrial Pollution Vehicular Pollution Area YName of plant E Area Area TPL* 506.81 g/ m3species A TPL* 308.02 TPL* 539.15 g/ m3 R g/m3 % Germination % % % % Germination Reduction Germination ReductionC. fistula S1 80 76 74 S2 92 80 76 S3 88 76 80 S4 92 68 84 A 86.0 3.16 75.0±2.64 12.79% 78.50 2.64 8.72%C. siamea S1 84 92 76 S2 92 76 80 S3 96 80 84 S4 92 84 92 A 91.0 2.64 83.0 3.16 8.79% 83.0 3.16 8.79%C. pulche-rrima S1 96 80 92 S2 92 68 84 S3 96 76 80 S4 88 84 81 A 93.0  2.44 77.0±3.16 17.20% 84.25 2.12 9.40%D. regia S1 90 70 83 S2 96 76 81 S3 84 68 83 S4 96 72 82 A 91.5±2.95 71.50±2.23 21.85% 82.25±+1.17 10.10%P. inerme S1 96 72 84 S2 80 68 76 S3 84 64 78 S4 88 76 82 A 87.0  3.16 70.0  2.82 19.54% 80.0  2.64 8.04%*TPL -Total Pollution Load, Sampling year - S1 –2002, S2 –2003, S3–2004 ; A – Average Values,Samples of 100 seeds each.3.3.2 Seed Weight The results presented in Table 3.2 and Fig. 3.1 clearly indicates a reduction in weight as compared to Low pollution area. Maximum reduction in seed weight was recorded in P. inerme (15.69%) followed by C. pulcherrima (12.45%) and minimum in C. fistula (10.00%) in Industrial Pollution Area. While in Vehicular Pollution Area maximum reduction was recorded in C. fistula (25.37%) followed by P. inerme (22.14%) and minimum in D. regia (8.45%). Obviously the seeds of P. inerme, C. pulcherrima and C. fistula affected more than other species due to air pollutants. 43
    • 3.3.3 Seed Density Seed density is an important characteristics related to seed quality and seed vigor. Seeds collected from polluted sites showed poor density. In this connection the air of IPA was found to be more damaging than VPA (Table 3.3, Fig. 3.2). As for as different species are concerned D. regia was more affected in both the sites as there was more than 21 % reduction noted, while minimum reduction was observed in IPA for C. pulcherrima, i.e. 13.95 and 0.79 for P. inerme at VPA.3.3.4 Seed Soundness The data pertaining to soundness of seeds are presented in Table 3.4 and Fig. 3.3. A seed lot with higher proportion of shriveled, under-sized, under- developed, discoloured and insect damage seed is considered as poor. It is evident that the percentage of unhealthy seeds in comparison to Low pollution area was higher in both the polluted sites. Higher proportion of poor seed was found in C.siamea, whereas lower proportion of poor seed was recorded in P.inerme IPA and VPA respectively (Fig. 3.1 to 3.5).3.3.5 Seed Germination The data presented in Table 3.3.5 and Fig. 3.5 clearly indicates that the seed collected from polluted areas germinated poorly than the reference area. It was also observed that the seeds of reference area germinated 10-15 days early than the seeds of polluted areas. In IPA maximum reduction in seed germination was observed in D. regia (21.85%) followed by P. inerme (19.54%) and minimum reduction was noted in 44
    • C. siamea (8.79%). In VPA also maximum reduction was observed in D. regia (10.10%) and C. pulcherrima (8.045%) respectively. Thus it is clear that D. regia affected more irrespective of pollution site indicating its sensitivity to air pollution. The maximum reduction in seedling growth was recorded in P. inerme, i.e. 5.01 % in VPA followed by D. regia, i.e. 3.29% in IPA (Table 3.6) and minimum reduction in seedling growth was recorded in C. fistula for VPA and P. inerme for IPA as compared to Low pollution area.3.4 DISCUSSION Air pollution resulting from industrial and urban development has resulted in localized and regional damage to plants. The main air pollutants toxic to plants are Ozone, Sulphur dioxide and Nitrogen dioxide. Presently Ozone is considered to cause more worldwide damage to vegetation than all the other air pollutants combined (Heagle 1989). There is ample evidence that gaseous air pollutants has adverse effects on leaves and total yield and that these effects vary enormously among genotype and environments. However, less research has been directed to the correlation of leaf injury and its associated photosynthesis impairment with air pollution effects on flowering, pollination and seed set and on carbon allocation from leaves to developing fruit (Ormord 1996). Significant reduction in fruit length, percent fruit setting and seed per fruit in C. siamea growing along road side in Agra has been recently reported by Chauhan et al. (2004) which further support the present finding. The present observations are supported by several workers (Dubey and Pawar 1985, Rout and Varshney 1996 and Awasthy 1998). According to these workers there is a mark reduction in fruit numbers, size, colour, quality and consumer acceptability in the presence of air pollutants and automobile exhaust. The phase of germination for a plant is an important aspect from its growth and production point of view. Seed collected from variously polluted sites were subjected to germination and vigour test showed reduction in these values. Similar observations have been noted in C. tora and C. occindentalis seeds 45
    • collected from automobile polluted sites by Krishnayya and Bedi (1986). Theyhave also reported reduced seed viability as a function of lead pollution.The present findings are also in confirmations with the work of Houston andDochinger (1979) and Murdy and Ragasdale (1980). Degradation of both seedquality and quantity in Wheat grown in SO2 polluted Industrial area has beenreported (Pawar 1982). Seed quality in terms of crude protein and oil contentwas reduced by elevated ozone level in ambient air (Ollerenshow et al. 1999).However the individual pods borne on their branches were heavier andcontained more seeds perhaps as a consequence of compensatory response(Ollerenshow et al. 1999).Reduced seed germination, less vigour and poor seedling growth observed in allthe study plants can be attributed to air pollutants prevailing in those sites fromwhere these samples had been collected. Early germination of seeds of referencearea in comparison to seeds of polluted sites can be a function of their goodhealth, high vigour and vitality, on account of their higher values of seed weightand density. This clearly indicates their good quality over polluted site seeds.Maximum reduction in seed germination in C. fistula and P. inerme indicatesthat pollutants affected seeds of these trees more. The maximum reduction ingermination of C. fistula seeds is in confirmation with the maximum loss inseed weight and seed density of the same plant. Thus it is clear that seeds of C.fistula were affected more by pollution than rest of tree species. Which isfurther confirmed through its poor seedling performance. All these alterationsmight have resulted due to the combined effect of mixture of gaseous as well asparticulate pollution on the vegetative as well as reproductive parts of the plantsduring the course of their developmental stages.Change in seed colour reduction in seed weight, seed density, soundness andgermination in plants growing in Industrial and Vehicular Pollution Areas canbe attributed to the prevailing air pollution in these sites from where the sampleswere collected. Obviously the change in seed colour is an indirect effect of airpollutants on them. During the course of development the seeds were notexposed directly to the pollutants as the fruits. Thus the change in colour of theseeds appears to be an indirect indication of seed quality. 46
    • On the basis of overall assessment of all the five tree species with reference totheir sensitivity to air pollution they can be arranged as:C. fistula > C. siamea > D. regia > P. inerme > C. pulcherrimaThus it is suggested that C. fistula is more sensitive to urban air pollution and C.pulcherrima the least, while D. regia is moderately affected. 47
    • ACKNOWLEDGEMENTWe gratefully acknowledge help extended by Dr. Dilip Wagela, Scientist, M.P.Pollution Control Board, Indore (India) and Professor Dr. Bholeshwar Dube, Head,Department of Botany, Mata Jijabai Girls P.G. College, Indore (India) in the course ofresearch and preparation of the book. 48
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