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Ambient lead (Pb) concentration, bioaccumulation and its effects on leaf epidermal structures of Bougainvillea spectabilis.

Introduction

Lead (Pb) the widely used element of the transition metals, had played various roles in the development of human civilization. The major source of Pb in the environment is the automobile exhaust fumes because of the addition of the lead in the form of tetra ethyl lead or tetra methyl lead to the gasoline as an antiknock substance in the last decades. Thus road traffic plays a major role in polluting the air of urban environment. In spite of all regulatory measures and total ban on the use of lead antiknock additives, it continues to be most serious global environmental problem as a major pollutant for both terrestrial and aquatic ecosystems. Excess lead causes a number of toxicity symptoms in plants as stunted growth, chlorosis, blackening of root system. Harmful effects of polluted air on plant as alterations in leaf epidermal structures have been studied by various workers (Salgare and Achereker, 1991; Rai and Kulshreshth, 2006). Some biochemical and ultra-structural changes have been reported in plant foliage exposed to auto-exhaust pollution by Verma and Singh, (2006). Malviya and Wagela (2001) had also estimated the Pb content in plant vegetation. Sharma and Dubey (2005) reviewed the morphological, physiological and biochemical effects of Pb toxicity on plants. Ji Tao et al. (2006) assessed the bioavailability, phytotoxicity and bioaccumulation of Pb in ryegrass and millet. Bougainvillea spectabilis is also a wild and ornamental shrub, which is widely used in green belt and social forestry to enhance the beauty and purity of roadside environment. Thus this paper also aims:

1.to estimate the ambient Pb concentration. 2. to estimate bioaccumulation of Pb by leaves of B. spectabilis. 3. to investigate the leaf epidermal structure of B. spectabilis and 4. to correlate with Pb present in air and leaves.

Materials and Methods

Air quality monitoring of ten selected sites (Sirmaur Chowk, Civil Lines, A.P.S. University campus (contorl site), Stadium Chowk, Dhobia Tanki area, Jaistambh chowk, PTS chowk, Nagar Nigam Building area, Prakash chowk, Saman chowk) have been carried out to assess the ambient Pb concentration along the major roadsides and national highways and crossings through the Rewa city. Air samples for Pb analysis were collected by Respirable Dust Sampler (Envirotech model APM 460-BL/APM 411) for eight hours in a day as per the standards of Central Pollution Control Board (CPCB), India. Leaf samples were also taken from the selected eleven sites.

Quantitative estimation of lead ([Pb.sup.++]): Quantification of [Pb.sup.++] in the air was done by the digestion of dust collected in the filter paper (GFA-3). To determine suspended metal ([Pb.sup.++]) the filter paper was digested with the help of nitric acid- perchloric acid. A little amount was taken for metal quantification. The concentration was analyzed by Atomic Absorption Spectrophotometer, Model PERKIN ELMER, MLA MLS 300 (1998-99) as per standards of CPCB, India.

Calculation: Concentration of heavy metals in the sample was calculated by comparing the sample minus blank reading of the calculation curve. [micro]g/[m.sup.3] heavy metal = CV/T, Where, C=mg/L Heavy metal in sample V=Sample Volume in ml, T=Total volume of air passed through the cyclone (m3).

Quantitative estimation of lead ([Pb.sup.++]) in leaves: Leaves are oven dried at 70 [degrees]C for 46 hours. 2-3 gm of finely grounded leaf material was taken in 100 ml flask and added the mixture of 25 ml nitric acid and 5 ml perchloric acid, digested by slow heating initially then fast heating until acid refluxes down the side of the flask. After cooling, transferred it to 50 ml. calibrated flask and make up volume with distilled water.

Calculation: mg/gm heavy metal=CV/w, Where, C=mg/L heavy metal in sample, V=Sample volume in ml, W=Weight of leaf sample in gm.

Microscopy or Anatomical Study: Leaf surface characteristics were studied with light microscope and Motic images camera. The leaf epidermal peel slides were made by the method of lasting Impressions. Leaf impression was examined under at least 400x magnification by light microscope and 400x, 1000x magnification by digital microscope. Number of stomata and epidermal cell were counted per square millimeter area and the stomatal frequency and stomatal index were calculated by using the following formulae. Stomatal frequency (S.F.) = S / E x 100, Stomatal Index (S.I.) = S /E + S x 100, Where S=Average no. of stomata, E=Average no. of Epidermal cells.

Cell size measurement: Size (Length & breadth) of epidermal cells and guard cells were measured with ocular micrometer under high power magnifications with the help of "stage-ocular micro-metery" and "Motic Images plus 2.0 ML" software.

Statistical Analysis were done by the "Graph Pad In stat 3" software, student's t-test and coefficient of correlation were applied for interpretation.

Results and Discussion

Lead contents in particulate matters of ambient air

Seasonal [Pb.sup.++] concentrations in ambient air at selected roadsides are shown in Table I. The highest Pb content was recorded at Sirmaur Chowk during winter months (0.4367 [micro]g/[m.sup.3]) and rainy (0.3758 [micro]g/[m.sup.3]) months, but in summer months at Jai Stambh Chowk (0.1386 [micro]g/[m.sup.3]). The lowest Pb content in the ambient air was recorded at UTD during winter (0.0542 [micro]g/[m.sup.3]), rainy (0.0324 [micro]g/[m.sup.3]) and summer (0.0295 [micro]g/[m.sup.3]) months. Seasonal average lead contents of particulate matters in the ambient air of Rewa city were computed from the basic data (Figure I). Results revealed higher lead contents during winter months (0.175 [micro]g/[m.sup.3]) followed by rainy (0.103 [micro]g/[m.sup.3]) and summer (0.073 [micro]g/[m.sup.3]) months. It is obvious from the results that Pb levels were higher during winter months followed by rainy and summer months. The similar trend of variation was also observed by Sayed et al., 2002 and Pradhan et al. (2004). Comparatively higher Pb contents estimated from the ambient air can be attributed to the traffic density of different types of auto-vehicles in operation. Wagela et al. (1998) and Malviya and Wagela (2001) have also reported similar findings in areas adjacent to the roadsides.

Automobile exhaust fumes results emission of lead in ambient air which ultimately settles down and gets mingled with roadside dust (Wagela, 1998). This study also follows the same trend having high lead concentration in the area where automobile pollution was high. Despite abatement on the use of Pb additives in the petrol fuels the presence of considerable amount of Pb concentration in the air suggests either of two concluding options. The first one is that the ban on the use of lead additives in petrol is not practiced honestly on ground level of execution and the second one is the recycling of Pb particulates settled down on the ground and arrived on the ground from other sources like battery discharging processes, continuous demolition of older buildings containing leaded paints (Montgomery and Mather, 2005; Farfel et al., 2005), which might be the another reason for the major source of inventory of Pb particulate emissions.

[FIGURE I OMITTED]

Various recent studies also support the second option of lead recycling by soil containing Pb in heavy amount, as the major source of inventory (Banat et al., 2005; Li, 2006; Nabulo et al., 2006).

Bioaccumulation of Lead (Pb): Table II shows the seasonal Pb content in the leaves of B. spectabilis growing at selected roadsides. Maximum accumulation of [Pb.sup.++] by the leaves of B. spectabilis was recorded at Sirmaur Chowk during winter (12.84 mg/g), rainy (10.99 mg/g) and summer (9.446 mg/g) seasons while the minimum was recorded for leaves at UTD campus during winter (2.034 mg/g), rainy (1.07 mg/g) and summer (1.001 mg/g). Seasonal average Pb contents of B. spectabilis leaves revealed highest Pb in the leaves during winter (5.013 mg/g) followed by rainy (3.636 mg/g) and summer (2.801 mg/g) months (Figure II). The correlation was calculated between these seasonal average values of leaf Pb and seasonal average Pb in ambient air which shows very strong positive correlation (r=0.9540). This study indicates the accumulation of substantial amount of lead in the leaves of B. spectabilis growing along roadsides. Presence of lead in the leaves of plant of urban areas is also conformed by various recent studies (Jaradat and Momani, 1999; Indukumari et al., 2005 Tomasevic et al, 2005; Swaileh et al, 2006; Oliva and Mingorance, 2006; Nabulo et al., 2006). Plant leaves have shown greater accumulation of Pb at sites with greater particulate contents in the ambient air (r=0.9540). The higher Pb was accumulated by the leaves of plants of high traffic area than those of low traffic area and control site. This increase could be due to prolonged exposure of plants to roadside exhaust of high traffic area and recycled lead dust (Indukumari et al., 2005). Swhaileh et al (2006) have reported Pb accumulation almost 3-4 times more in old leaves than new ones. This is perhaps one of the reasons for higher Pb contents in the leaves during winter months.

[FIGURE II OMITTED]

Effects on Bougainvillea spectabilis: Figure. III depicts the healthy as well as damaged epidermal and guard cells of Bougainvillea spectabilis at control and polluted zones, respectively.

Guard cells: At ventral surface no significant changes have been observed in guard cell lengths during winter months. However a significant increase was observed in width at most of the sites. The stomatal number, stomatal frequency and stomatal index showed decreasing pattern. A significant increase was observed in cell length and width at most of sites during summer and rainy months. Decrease in number of stomata has been observed during summer while increased number of stomata was noticed in rainy months. Stomatal frequency and stomatal index were notably increased than control site during both the seasons (Table-III and Table-IV).

Epidermal cells: Table-V and Table-VI presents the alterations in the cell length and width of epidermal cells. At dorsal surface the epidermal cell length exhibited significant decrease for some sites during winter months. There was significant decrease in epidermal cell width at most of sampling sites as compared to control plants during winter month and rainy months. However significant increase in cell width was noticed for most of sites during summer months. Leaves of sampling sites registered increased number of epidermal cells at all sites except two during winter months. The epidermal cell numbers were found to be increased on dorsal surface. In rainy season the number of epidermal cells was increased in all sites except three sampling sites as compared to control. Although, reduction in epidermal cell size was not observed in all seasons at dorsal and ventral surfaces some of the individual cells were in highly ruptured, degenerated and deteriorated conditions in all samples. Correlation was established between Pb contents and anatomical features of leaves with respect to three seasons (Table VII). Results revealed increase in length and width of guard cells of ventral surface (winter samples) and decreased width of rainy samples were negatively correlated with leaf Pb contents. Similarly, there was negative correlation between lead contents and epidermal cells size of both the surface of leaves, except on dorsal surface of summer samples and ventral surface of monsoon samples. The number of stomata and epidermal cells were also negatively correlated with Pb contents during summer months (Table VIII). These negative correlations indicate the negative effect of lead contents on cell size. Raina and Sharma (2006) have also observed marked alterations in the size and number of foliar epidermal and guard cells, stomatal frequency and stomatal index in both abaxial and adaxial surfaces of leaves of B.spectabilis as air pollution effects. Verma and Singh (2006) have reported the similar results regarding effect of urban air pollution on leaf epidermal parameters.

[FIGURE III OMITTED]

The significant reduction observed in cell size of B. spectabilis may be attributed to Pb toxicity, similarly reduced number of stomata per square millimeter, stomata frequency and stomata index could be attributed to Pb toxicity which is supported by Wehyszko-Chmielwska et al. (2005). However the significant increase in cell size may be attributed to the physiological and anatomical adaptive measures adopted by the plants naturally, if the no. of stomata will reduce then the guard cell size will increase at same surface. Whether the number of stomata reduction at dorsal surface leads the increased no. of stomata, stomata frequency and stomata index at ventral surface (Hethrington and Woodward, 2003).

Conclusion and Recommendations

From the present study it may be concluded that ambient Pb concentration is in considerable amount at roadside air. Pb in less than the permissible limit may cause toxicity in the plants and human by prolonged exposure. Despite the higher accumulation of [Pb.sup.++] by leaves of B. spectabilis. The plant shows much resistant nature through the less reduction observed in leaf epidermal structures due to [Pb.sup.++] toxicity. So it can be recommended for continuous use for roadside plantation as pollution indicators and for phytoremediation of Pb. Further investigation is also needed regarding other physiological effects (especially photosynthesis) of the plant as cell deterioration and alterations are found to be significant in the study.

References

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Mani Singh * and R. M. Mishra

School of Environmental Biology, Awdhesh Pratap Singh University, Rewa (M.P.) India

* E-mail: smani801@gmail.com
Table I: Pb content in Ambient Air of Rewa City ([micro]g/[m.sup.3]).

 Winter Summer Rainy

Stadium Chowk 0.1723 0.0528 0.1166
Sirmaur chowk 0.4367 0.1063 0.3758
UTD 0.0542 0.0295 0.0324
Saman chowk 0.1924 0.1013 0.1190
PTS chowk 0.0695 0.0513 0.0360
Dhobia Tanki 0.1783 0.0633 0.0565
Civil Lines 0.0761 0.0439 0.0531
Jaistambh chowk 0.3688 0.1386 0.1417
Nagar Nigam 0.1025 0.0769 0.0417
Prakash chowk 0.0992 0.0635 0.0599

Table II: Pb content in the leaves of B. spectabilis (mg/g).

 Winrer Rainy Summer

Stadium Chowk 4.001 2.205 1.066
Sirmaur chowk 12.846 10.991 9.446
UTD 2.034 1.078 1.001
Saman chowk 4.552 3.020 1.999
PTS chowk 2.878 1.894 1.031
Dhobia Tanki 4.471 3.121 2.543
Civil Lines 3.097 2.795 1.545
Jaistambh chowk 11.083 9.637 8.771
Nagar Nigam 3.894 1.843 1.252
Prakash chowk 3.551 1.830 1.077
Transport Nagar 2.744 1.588 1.082

Table III: Average length and width of Guard cells, number of
stomata, stomatal frequency and stomatal index of Bougainvillea
spectabilis growing at different sites of Rewa city.

 SITES
 Stadium Sirmaur
SEASONS Chowk chowk

WINTER DORSAL L 66.85 65.46
 [+ or -] 12.531 [+ or -] 6.954

 W 15.67 17.25
 [+ or -] 3.525 [+ or -] 3.577

 NFS 10 30

 SF 1.818 3.030

 SI 1.785 2.941

 VENTRAL L 66.37 61.86
 [+ or -] 8.520 [+ or -] 9.997

 W 18.25 15.27
 [+ or -] 4.259 [+ or -] 3.084

 NFS 30 170

 SF 3.659 15.740

 SI 3.529 13.60

SUMMER DORSAL L 64.51 67.242
 [+ or -] 7.512 [+ or -] 9.222

 W 12.67 13.48
 [+ or -] 3.1320 [+ or -] 3.916

 NFS 25 30

 SF 4.464 25

 SI 4.273 20

 VENTRAL L 67.23 69.15
 [+ or -] 6.452 [+ or -] 4.46
 W 13.00 13.95
 [+ or -] 3.001 [+ or -] 3.121

 NFS 145 160

 SF 19.333 22.222

 SI 16.201 18.182

RAINY DORSAL L 59.867 67.86
 [+ or -] 11.273 [+ or -] 12.480

 W 13.97 12.21
 [+ or -] 4.269 [+ or -] 4.837

 NFS 25 40

 SF 2.631 2.564

 SI 2.564 2.00

 VENTRAL L 75.09 83.58
 [+ or -] 9.574 [+ or -] 13.294

 W 14.37 18.2
 [+ or -] 3.007 [+ or -] 3.282
 NFS 135 140

 SF 1.157 10.935

 SI 10.037 9859

 SITES
 Saman PTS
SEASONS chowk Chowk

WINTER DORSAL L 6589 58.02
 [+ or -] 11.24 [+ or -] 6.451

 W 14.5 12.22
 [+ or -] 3.546 [+ or -] 5.123

 NFS 20 20

 SF 4 2.903

 SI 3.846 2.821

 VENTRAL L 6332 62.15
 [+ or -] 7.55 [+ or -] 8456

 W 1794 18.67
 [+ or -] 4.894 [+ or -] 4.568

 NFS 60 110

 SF 7.407 14.102

 SI 6.896 12.359

SUMMER DORSAL L 62.03 48.24
 [+ or -] 7.571 [+ or -] 6.879

 W 1298 11.89
 [+ or -] 3.193 [+ or -] 2787

 NFS 30 47

 SF 2.727 5.341

 SI 2.655 5.070

 VENTRAL L 6828 63.456
 [+ or -] 7.727 [+ or -] 7.46

 W 13.78 10.33
 [+ or -] 3.002 [+ or -] 3.521

 NFS 150 160

 SF 21.127 18.823

 SI 17.442 15.841

RAINY DORSAL L 60.48 59.65
 [+ or -] 6.789 [+ or -] 5.48

 W 14.78 13.50
 [+ or -] 5.456 [+ or -] 4.234

 NFS 29 20

 SF 3.258 2.705

 SI 3.156 2.062

 VENTRAL L 7789 50.49
 [+ or -] 8.486 [+ or -] 7.685

 W 1489 12.37
 [+ or -] 4.56 [+ or -] 3.481

 NFS 145 132

 SF 11.837 10.688

 SI 10.584 9.656

 SITES
 Dhoba Tanki Civil
SEASONS Lines

WINTER DORSAL L 66.54 61.15
 [+ or -] 13.604 [+ or -] 12.465

 W 13.07 14.54
 [+ or -] 1.871 [+ or -] 2.154

 NFS 40 55

 SF 5.00 6.548

 SI 4.762 6.145

 VENTRAL L 65.84 65.46
 [+ or -] 14.157 [+ or -] 8.02

 W 13.05 15.95
 [+ or -] 3.345 [+ or -] 13.14

 NFS 130 135

 SF 13.131 15.517

 SI 11.607 13.433

SUMMER DORSAL L 65.35 53.46
 [+ or -] 5.781 [+ or -] 6.452

 W 13.00 12.02
 [+ or -] 4.632 [+ or -] 3.156

 NFS 30 50

 SF 3.158 5.464

 SI 3.061 5.181

 VENTRAL L 67.75 60.35
 [+ or -] 6.482 [+ or -] 4.453

 W 12.38 11.26
 [+ or -] 3.45 [+ or -] 5.461

 NFS 155 156

 SF 19.645 15.757

 SI 16.419 13.612

RAINY DORSAL L 59.56 61.45
 [+ or -] 6.15 [+ or -] 9.79

 W 13.08 15.53
 [+ or -] 4.153 [+ or -] 3.478

 NFS 30 35

 SF 3.030 4.242

 SI 2.941 4.070

 VENTRAL L 79.86 58.78
 [+ or -] 8.795 [+ or -] 9.78

 W 14.89 14.9
 [+ or -] 2.451 [+ or -] 3.846

 NFS 138 134

 SF 11.500 11.260

 SI 10.314 10.121

 SITES

 Jaistambh Nagar Nigam
SEASONS chowk

WINTER DORSAL L 63.38 76.245
 [+ or -] 11.928 [+ or -] 10.488

 W 14.37 14.89
 [+ or -] 3.441 [+ or -] 3.480

 NFS 40 45

 SF 4.933 4.438

 SI 4.706 4.249

 VENTRAL L 63.53 62.45
 [+ or -] 11.05 [+ or -] 10.46

 W 18.44 16.45
 [+ or -] 4.551 [+ or -] 4.346

 NFS 140 115

 SF 14.286 11.386

 SI 12.50 10.222

SUMMER DORSAL L 67.542 63.468
 [+ or -] 7.251 [+ or -] 5.780

 W 13.23 12.45
 [+ or -] 2.96 [+ or -] 2.461

 NFS 30 43

 SF 2.5 4.3

 SI 2.439 4.122

 VENTRAL L 68.46 66.57
 [+ or -] 5.467 [+ or -] 5.461

 W 13.85 13.01
 [+ or -] 3.67 [+ or -] 4.43

 NFS 140 148

 SF 19.774 17.011

 SI 17.327 14.538

RAINY DORSAL L 58.58 61.46
 [+ or -] 9.051 [+ or -] 8.184

 W 11.27 14.95
 [+ or -] 2.326 [+ or -] 3.485

 NFS 20 27

 SF 2 2.903

 SI 1.961 2.821

 VENTRAL L 67.67 69.764
 [+ or -] 10.855 [+ or -] 12.463

 W 14.58 14.46
 [+ or -] 2.508 [+ or -] 4.45

 NFS 170 140

 SF 20 14.141

 SI 16.667 12.389

 SITES
 Prakash Transport
SEASONS chowk Nagar

WINTER DORSAL L 73.12 55.36
 [+ or -] 10.928 [+ or -] 9.656

 W 16.78 12.56
 [+ or -] 3.729 [+ or -] 4.098

 NFS 50 40

 SF 4.167 3.448

 SI 4.000 3.333

 VENTRAL L 6697 67.01
 [+ or -] 8.941 [+ or -] 8.622

 W 14 19.84
 [+ or -] 3.383 [+ or -] 5.169

 NFS 80 130

 SF 7.692 17.105

 SI 7.1429 14.607

SUMMER DORSAL L 62.00 48.89
 [+ or -] 4.245 [+ or -] 7.546

 W 12.06 11.23
 [+ or -] 3.156 [+ or -] 3.15

 NFS 24 18

 SF 1.846 1.488

 SI 1.813 1.464

 VENTRAL L 62.48 61.45
 [+ or -] 7.462 [+ or -] 5.451

 W 12.48 11.46
 [+ or -] 3.123 [+ or -] 3.124

 NFS 156 165

 SF 17.528 19.527

 SI 14.914 16.337

RAINY DORSAL L 61.489 60
 [+ or -] 10.79 [+ or -] 14.561

 W 1521 13.84
 [+ or -] 5.51 [+ or -] 3.542

 NFS 30 24

 SF 3.261 2.667

 SI 3.158 2.597

 VENTRAL L 67.75 51.01
 [+ or -] 11.46 [+ or -] 13.592

 W 13.46 12.46
 [+ or -] 3.48 [+ or -] 4.453

 NFS 135 133

 SF 13.500 11.083

 SI 11.894 9.977

 SITES UTD

SEASONS

WINTER DORSAL L 59.17
 [+ or -] 11.047

 W 12.8
 [+ or -] 2.06

 NFS 75

 SF 10.949

 SI 9.868

 VENTRAL L 64.02
 [+ or -] 5.160

 W 13.02
 [+ or -] 3.625

 NFS 170

 SF 20

 SI 16.667

SUMMER DORSAL L 50.24
 [+ or -] 8.967

 W 10.64
 [+ or -] 1.449

 NFS 70

 SF 7.778

 SI 7.216

 VENTRAL L 57.6
 [+ or -] 5.06

 W 9.7
 [+ or -] 1.703

 NFS 185

 SF 15.417

 SI 13.357

RAINY DORSAL L 63.57
 [+ or -] 8.949

 W 16.74
 [+ or -] 2.748

 NFS 40

 SF 5

 SI 4.762

 VENTRAL L 54
 [+ or -] 8.540

 W 14.3
 [+ or -] 3.293

 NFS 130

 SF 11.077

 SI 9.367

Table IV: Values of 't' test between size of guard cells of
Bougainvillea spectabilis leaves of polluted and control sites.

SITES Stadium Sirmaur Saman PTS
 Chowk Chowk Chowk Chowk
SEASONS

WINTER DORSAL LGC 1.454 1.524 1.348 0.284
 P=0.1632 P=0.1449 P=0.1943 P=0.7794

 WGC 2.223 * 3.409 ** 1.311 0.332
 P=0.0393 P=0.0031 P=0.2064 P=0.7436

 VENTRAL LGC 0.746 0.607 0.242 0.597
 P=0.4653 P=0.5513 P=0.8115 P=0.5580

 WGC 2.957 ** 1.495 2.555 3.064 **
 P=0.0084 P=0.1522 0.0199 P=0.0067

SUMMER DORSAL LGC 3.858 ** 4.180 *** 3.177 ** 0.560
 P=0.0012 P=0.0006 P=0.0052 P=0.5826

 WGC 1.860 * 2.151 * 2.110 * 1.258
 P=0.0793 P=0.0453 P=0.0491 P=0.2243

 VENTRAL LGC 3.714 ** 5 415 *** 3.657 ** 2.054 *
 P=0.0016 P<0.0001 P=0.0018 P=0.0548

 WGC 3.024 ** 3.780 ** 3.738 ** 0.509
 P=0.0073 P=0.0014 P=0.0015 P=0.6167

RAINY DORSAL LGC 0.816 0.883 0.870 1.181
 P=0.4265 P=0.8834 P=0.3958 P=0.2529

 WGC 1.725 2.575 * 1.015 2.030 *
 P=0.1016 P=0.0191 P=0.3237 P=0.0574

 VENTRAL LGC 5 198 *** 5.920 *** 6.275 *** 0.9661
 P<0.0001 P<0.0001 P<0.0001 P=0.3468

 WGC 0.050 2.653 * 0.332 1.274
 P=0.9610 P=0.0127 P=0.7439 P=0.2190

SITES Dhobia Civil Jaistambh
 Tanki Lines Chowk
SEASONS

WINTER DORSAL LGC 1.330 0.3759 0.819
 P=0.2002 P=0.7114 P=0.4236

 WGC 0.307 1.846 * 1.238
 P=0.7625 P=0.0814 P=0.2316

 VENTRAL LGC 0.382 0.809 0.127
 P=0.7070 P=0.4290 P=0.9003

 WGC 0.192 0.678 2.946 **
 P=0.9849 P=0.6797 P=0.0086

SUMMER DORSAL LGC 4.479 *** 0.922 4.745 ***
 P=0.0003 P=0.3688 P=0.0002

 WGC 1.538 1.257 2.485 *
 P=0.1415 P=1.421 P=0.0230

 VENTRAL LGC 3.903 ** 1.290 4.610 ***
 P=0.0010 P=0.2133 P=0.0002

 WGC 2.203 * 0.862 3.244 **
 P=0.0409 P=0.3998 P=0.0045

RAINY DORSAL LGC 1.168 0.505 1.240
 P=0.2581 P=0.6194 P=0.2310

 WGC 2.324 0.863 4.805
 P=0.0320 P=.3994 P=0.0001

 VENTRAL LGC 6.671 *** 1.164 3.130 **
 P<0.0001 P=0.2595 P=0.0058

 WGC 0.454 0.3747 0.2139
 P=0.6549 P=0.7122 P=0.8330

SITES Nagar Prakash Transport
 Nigam Chowk Nagar
SEASONS

WINTER DORSAL LGC 3.545 ** 2.839 * 0.821
 P = 0.0023 P=0.0109 P=0.4223

 WGC 1.634 2.954 ** 0.165
 P = 0.1196 P=0.0085 P=0.8704

 VENTRAL LGC 0.426 0.904 0.941
 P = 0.6754 P=0.3781 P=0.3592

 WGC 1.917 * 0.625 3.416 **
 P = 0.0713 P=0.5398 P=0.0031

SUMMER DORSAL LGC 3.921 ** 3.748 ** 0.364
 P = 0.0010 P=0.0015 P=0.7199

 WGC 2.004 * 1.293 0.538
 P = 0.0603 P=0.2123 P=0.5971

 VENTRAL LGC 3.810 ** 1.712 1.637
 P = 0.0013 P=0.1041 P=0.1190

 WGC 2.205 * 2.471 * 1.564
 P = 0.0407 P=0.0237 P=0.1352

RAINY DORSAL LGC 0.550 0.469 0.660
 P=5889 P=0.6444 P=0.5173

 WGC 1.275 0.786 2.046 *
 P = 0.2184 P=0.4422 P=0.0557

 VENTRAL LGC 3.300 ** 3.042 ** 0.589
 P = 0.0040 P=0.0070 P=0.5632

 WGC 0.09140 0.554 1.051
 P = 0.9282 P=0.5861 P=0.3073

LGC = Length of guard cells

WGC = Width of guard cell

*** = extremely significant

** = Very significant

* = Significant

t' value at 18 d.f. on 0.05% level is 1.734

Table V: Average length ([micro]m), width ([micro]m) and number of
epidermal cells of Bougainvillea spectabilis growing at different
sites of Rewa city.

SITES Stadium Sirmaur
 Chauk chauk
SEASONS

WINTER DORSAL L 87.78 81.76
 [+ or -] 2.589 [+ or -] 13.064

 W 58.75 51.48
 [+ or -] 18.142 [+ or -] 9.644

 NEC 550 990

 VENTRAL L 65.6 76.03
 [+ or -] 12.624 [+ or -] 17.354

 W 45.64 45.73
 [+ or -] 8.813 [+ or -] 7.330

 NEC 820 1080

SUMMER DORSAL L 94.61 95.6
 [+ or -] 11.632 [+ or -] 13.133

 W 71.34 60.83
 [+ or -] 6.554 [+ or -] 7.667

 NEC 560 120

 VENTRAL L 85.55 78.13
 [+ or -] 10.48 [+ or -] 8.179

 W 42.15 40.001
 [+ or -] 6.157 [+ or -] 6.546

 NEC 750 720

RAINY DORSAL L 100.99 94.55

 [+ or -] 17.503 [+ or -] 20.960

 W 79.36 58.28
 [+ or -] 15.292 [+ or -] 7.109

 NEC 950 1560

 VENTRAL L 70.56 95.94
 [+ or -] 20.421 [+ or -] 12.435

 W 48.59 50.94
 [+ or -] 11.247 [+ or -] 8.523

 NEC 1210 1280

SITES Saman chauk PTS chauk

SEASONS

WINTER DORSAL L 84.55 90.01
 [+ or -] 6.846 [+ or -] 15.32

 W 56.2 61.42
 [+ or -] 12.54 [+ or -] 10.654

 NEC 500 689

 VENTRAL L 64.49 75.12
 [+ or -] 14.54 [+ or -] 12.512

 W 44.68 48.68
 [+ or -] 8.243 [+ or -] 8.117

 NEC 810 780

SUMMER DORSAL L 96.61 88.45
 [+ or -] 14.117 [+ or -] 8.46

 W 72.23 60.31
 [+ or -] 14.063 [+ or -] 5.452

 NEC 1100 880

 VENTRAL L 87.86 74.64
 [+ or -] 14.015 [+ or -] 7.89

 W 40.29 46.546
 [+ or -] 7.401 [+ or -] 7.85

 NEC 710 850

RAINY DORSAL L 99.79 97.46
 [+ or -] 16.45
 W [+ or -] 14.89 60.476
 78.94 [+ or -] 14.5

 NEC [+ or -] 6.486 950
 890
 VENTRAL L 71.59 68.987
 [+ or -] 15.49 [+ or -] 16.458

 W 47.46 48.08
 [+ or -] 7.164 [+ or -] 8.146

 NEC 1225 1235

SITES Dhoba Tanki Civil Lines

SEASONS

WINTER DORSAL L 98.35 98.01
 [+ or -] 23.343 [+ or -] 20.45

 W 61.77 61.544
 [+ or -] 11.007 [+ or -] 12.346

 NEC 800 840

 VENTRAL L 91.77 87.56
 [+ or -] 17.629 [+ or -] 12.004

 W 56.92 55.413
 [+ or -] 10.517 [+ or -] 9.153

 NEC 990 870

SUMMER DORSAL L 95.34 88.79
 [+ or -] 11.53 [+ or -] 13.96

 W 65.12 59.06
 [+ or -] 7.54 [+ or -] 7.24

 NEC 950 915

 VENTRAL L 79.456 83.79
 [+ or -] 11.45 [+ or -] 4.56

 W 42.5 50.45
 [+ or -] 5.489 [+ or -] 5.79

 NEC 789 990

RAINY DORSAL L 99.59 98.578
 [+ or -] 4.95 [+ or -] 12.456

 W 70.99 75.48
 [+ or -] 15.78 [+ or -] 11.46

 NEC 990 825

 VENTRAL L 70.998 82.499
 [+ or -] 14.879 [+ or -] 13.48

 W 47.89 50.48
 [+ or -] 6.789 [+ or -] 8.978

 NEC 1200 1190

SITES Jastambh Nagar Ngam
 chauk
SEASONS

WINTER DORSAL L 88.61 85.94
 [+ or -] 20.73 [+ or -] 12.548
 W 68.53 60.132
 [+ or -] 9.972 [+ or -] 8.456

 NEC 810 1014

 VENTRAL L 79.42 80.59
 [+ or -] 13.993 [+ or -] 14.243

 W 46.65 46.02
 [+ or -] 10.286 [+ or -] 6.487

 NEC 980 1010

SUMMER DORSAL L 96.61 93.25
 [+ or -] 3.342 [+ or -] 5.642

 W 73.68 71.34
 [+ or -] 6.45 [+ or -] 4.58

 NEC 1200 1000

 VENTRAL L 78.53 80.67
 [+ or -] 7.458 [+ or -] 5.468
 W 39.97 44.56
 [+ or -] 8.486 [+ or -] 4.792
 NEC 708 870

RAINY DORSAL L 100.75 99.09
 [+ or -] 12.542 [+ or -] 13.489

 W 63.1 79.86
 [+ or -] 16.081 [+ or -] 12.245

 NEC 1000 930

 VENTRAL L 80.17 80.45
 [+ or -] 9.988 [+ or -] 12.489

 W 51.36 52.4536
 [+ or -] 12.257 [+ or -] 7.798

 NEC 850 990

SITES Prakash Transport
 chauk nagar
SEASONS

WINTER DORSAL L 80.97 91.28
 [+ or -] 12.209 [+ or -] 19.689

 W 56.17 61.6
 [+ or -] 12.851 [+ or -] 12.382

 NEC 1200 1160

 VENTRAL L 78.85 73.95
 [+ or -] 12.091 [+ or -] 12.205

 W 46.65 50.17
 [+ or -] 10.286 [+ or -] 8.503

 NEC 1040 760

SUMMER DORSAL L 92.12 90.45
 [+ or -] 7.46 [+ or -] 9.43

 W 70.76 59.89
 [+ or -] 8.67 [+ or -] 6.43

 NEC 1300 1210

 VENTRAL L 79.97 81.48
 [+ or -] 4.751 [+ or -] 9.465

 W 48.45 45.54
 [+ or -] 10.46 [+ or -] 6.481

 NEC 890 845

RAINY DORSAL L 98.89 99.458
 [+ or -] 11.463 [+ or -] 13.457

 W 79.89 64.45
 [+ or -] 15.45 [+ or -] 8.49

 NEC 920 900

 VENTRAL L 81.49 69.987
 [+ or -] 10.468 [+ or -] 11.484

 W 52.576 49.99
 [+ or -] 10.479 [+ or -] 11.121

 NEC 1000 1200

SITES UTD

SEASONS

WINTER DORSAL L 98.01
 [+ or -] 19.622

 W 73.43
 [+ or -] 13.463

 NEC 685

 VENTRAL L 85.17
 [+ or -] 34.703

 W 56.42
 [+ or -] 14.987

 NEC 850

SUMMER DORSAL L 84.89
 [+ or -] 10.534

 W 57.46
 [+ or -] 8.017

 NEC 900

 VENTRAL L 82.64
 [+ or -] 10.45

 W 56.87
 [+ or -] 6.492

 NEC 1200

RAINY DORSAL L 97.229
 [+ or -] 22.720

 W 80.83
 [+ or -] 27.124

 NEC 800

 VENTRAL L 86.9
 [+ or -] 13.087

 W 58.53
 [+ or -] 6.889

 NEC 1180

Table VI: Values of 't' test between size of epidermal cells of
Bougainvillea spectabilis leaves of polluted and control sites.

 SITES Stadium Sirmaur Saman
 Chowk Chowk Chowk
SEASONS
WINTER DORSAL LEC 1.634 2.180 * 2.048 *
 P=0.1195 P=0.0428 P=2.048

 WEC 2.055 * 4.191 *** 2.961 **
 P=0.0547 P=0.0005 P=0.0084

 VENTRAL LEC 1.676 0. 7449 1.738 *
 P=0.1110 P=0.4659 P=0.0993

 WEC 1.961 * 2. 026 * 2.171 *
 P=0.0656 P=0.0578 P=0.0436

SUMMER DORSAL LEC 1.959 2.012 * 2.104 *
 P=0.0658 P=0.0595 P=0.0497

 WEC 4.239 *** 0.961 2.885 **
 P=0.0005 P=0.3494 P=0. 0099

 VENTRAL LEC 0.622 1.075 0.944
 P=0.5419 P=0.2967 P=0.3576

 WEC 5.203 *** 5.786 *** 5.326 ***
 P<0.0001 P<0.0001 P<0.0001

RAINY DORSAL LEC 0.415 0.274 0.298
 P=0.6833 P=0.7872 P=7690

 WEC 0.149 2.543 * 0.214
 P=0.8830 P=0.0204 P=0. 8327

 VENTRAL LEC 2.130 * 1.584 2.388 *
 P=0.0472 P=0.1307 P=0.0281

 WEC 2. 383 * 2. 190 * 3.522 **
 P=0.0284 P=0.0419 P=0.0024

 SITES PTS Chowk Dhobia Tanki Civil Lines

SEASONS
WINTER DORSAL LEC 1.016 0.035 0.024
 P=0.3230 P=0.9723 P=0.9807

 WEC 2.212 * 2.120 * 1.019
 P=0.0401 P=0.0481 P=0.3217

 VENTRAL LEC 0. 865 0.536 0. 2058
 P=0.4003 P=0.5984 P=0.8392

 WEC 1.436 0.086 0.1813
 P=0.1681 P=0.9321 P=0.85 81

SUMMER DORSAL LEC 0.833 2.116 * 0.705
 P=0.4156 P=0.0485 P=0.4897

 WEC 0. 929 2.201 * 0.468
 P=0.3649 P=0.0410 P=0.6451

 VENTRAL LEC 1.932 * 0.649 0.319
 P=0.0693 P=0.5242 P=0.7534

 WEC 3.205 ** 5 345 *** 2.334 *
 P=0.0049 P<0.0001 P=0.0314

RAINY DORSAL LEC 0. 026 0.321 0.165
 P=0.9795 P=0.7518 P=0.8711

 WEC 2.093 * 0.992 0.575
 P=0.0508 P=0.3345 P=0.5727

 VENTRAL LEC 2.694 * 2.538 * 0.741
 P=0.0148 P=0. 0206 P=0.4684

 WEC 3.098 ** 3 479 ** 2. 249 *
 P=0.0062 P=0.0027 P=0.0372

 SITES Jaistambh Nagar Prakash
 Chowk Nigam Chowk
SEASONS
WINTER DORSAL LEC 1.041 1.639 2.332 *
 P=0.3115 P=0.1186 P=0.0315

 WEC 0.925 2.645 * 2.933 **
 P=0.3673 P=0.0165 P=0.0089

 VENTRAL LEC 0.4859 0.386 0.544
 P=0.6329 P=0.7040 P=0.5932

 WEC 1.700 2.014 * 1.700
 P=0.1064 P=0.0592 P=0.1064

SUMMER DORSAL LEC 3.354 ** 2.212 * 1.771
 P=0.0035 P=0.0401 P=0.0935

 WEC 4.985 *** 4.754 *** 3.562 **
 P<0.0001 P=0.0002 P=0.0022

 VENTRAL LEC 1.012 0.528 0.735
 P=0.3248 P=0.6038 P=0.4715

 WEC 5.002 *** 4.824 *** 2.163 *
 P<0.0001 P<0.0001 P=0.0443

RAINY DORSAL LEC 0.429 0.223 0.206
 P=0.6730 P=0.8263 P=0.8388

 WEC 1.778 * 0.1031 0.095
 P=0.0923 P=0.9215 P=0.9252

 VENTRAL LEC 1.293 1.128 1.021
 P=0.2125 P=0.2743 P=0.3209

 WEC 1.613 1.847 * 1.501
 P=0.1242 P=0.0811 P=0.1506

 SITES Transport
 Nagar
SEASONS
WINTER DORSAL LEC 0.766
 P=0.4538

 WEC 2.045 *
 P=0.0557

 VENTRAL LEC 0.964
 P=0.3476

 WEC 1.147
 P=1.147

SUMMER DORSAL LEC 1.224
 P=0.2296

 WEC 0.748
 P=0.4643

 VENTRAL LEC 0.260
 P=0.7977

 WEC 3.906 **
 P=0.0010

RAINY DORSAL LEC 0.267
 P=0.7926

 WEC 1.822 *
 P=0.085

 VENTRAL LEC 3.072 **
 P=0.0066

 WEC 2.064 *
 P=0.0537

LEC=Length of epidermal cells

WEC=Width of epidermal cell

*** = extremely significant

** = Very significant

* = Significant

t' value at 18 d.f. on 0.05% level is 1.734

Table VII: Values of Correlation coefficient (r) between Pb content
and numbers of stomata and epidermal cells of Bougainvillea
spectabilis (Willd.) leaves.

 Stomata Epidermal cells

 Dorsal Ventral Dorsal Ventral

Winter -0.064 0.512 0.0384 0.584
Summer -0.131 -0.224 -0.422 -0.588
Rainy 0.294 0.655 * 0.783 * -0.161

Table VIII: Values of Correlation coefficient (r) between Pb content
and size of guard cells and epidermal cells of Bougainvillea
spectabilis (Willd.) leaves.

 Guard cells Epidermal cells

 Dorsal surface Ventral surface Dorsal surface

 Length Width Length Width Length Width

Winter 0.04791 0.4723 0.4921 -0.0944 -0.3604 -0.1781
Summer 0.5623 0.7162 0.5920 0.6176 0.5602 0.05328
Rainy 0.5106 -0.7810 ** 0.4457 0.7335 * -0.4125 -0.5928

 Epidermal cells

 Ventral surface

 Length Width

Winter -0.0024 -0.3278
Summer -0.3311 -0.6265 *
Rainy 0.6938 * 0.2382

* Significant at 0.05 % (8 d. f.).
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Author:Singh, Mani; Mishra, R.M.
Publication:International Journal of Applied Environmental Sciences
Date:May 1, 2010
Words:6941
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