NUS ANT AR A BIOS C IENC E
ISSN: 2087-3948
E-ISSN: 2087-3956
DOI: 10.
13057/nusbiosci/n100206 Vol.
No.
2, pp.
May 2018 Seeds characters, pollen fertility and flavonoids of ten Brassicaceae collected near a kilns thermal power plant for air pollution bioindication MITRA NOORI1,ou.
ZAHRA BAGHAEIFAR2.
ABDOLKARIM CHEHREGANI3.
FARZANEH FARAKI2 ou Department of Biology.
Faculty of Science.
Arak University.
Arak 38156-8-8349-Iran.
Tel.
/fax.
: 98-86-34173406, email: m-noori@araku.
Department of Biology.
Hamadan Payame Noor University.
Hamadan 65199-9951.
Iran Department of Biology.
Faculty of Science.
Bu-Ali Sina University.
Hamadan 6517838695.
Iran Manuscript received: 9 April 2018.
Revision accepted: 13 May 2018.
Abstract.
Noori M.
Baghaeifar Z.
Chehregani A.
Faraki F.
Seeds characters, pollen fertility and flavonoids of ten Brassicaceae collected near a kilns thermal power plant for air pollution bioindication.
Nusantara Bioscience 10: 96-104.
Shazand Steam Power Plant located on North-East of Shazand.
Iran began to work from 2000.
The power plant necessity fuel is natural gas and mostly heavy fuel oil.
The most pollutant of power plant is sulfur compounds in addition to nitric and carbon mono oxide.
Because environmental pollutants influence plant fertility and chemical compounds, therefore this study was done on ten wild Brassicaceae (Alyssum linifolium linifolium.
Alyssum longistylum.
Alyssum marginatum.
Choriospora persica.
Clypeola lappacea.
Conringia perfoliata.
Descurainia sophia.
Goldbachia laevigata.
Isatis kotschyana and Neslia apiculat.
taxa collected from the thermal power plant area for bioindication of regional air pollution comparing to controls collected 40 km away from the power plant.
Brassicaceae members are important for their ecological, pastoral, medicinal and edible points.
Seed width and length max.
and their ratio and abnormal seed percentage were Pollen abnormality and sterility percentages determined using MuntezingAos acetocarmine and light microscopy.
Also, their pollen flavonoids were semi-quantitatively assessed using two-dimensional paper and thin layer chromatography.
Results showed seeds health and their dimensions reduction in polluted samples in comparison with controls.
In C.
lappacea significant differences of seed and pollen abnormality and pollen sterility percentages, morin and kaempferol concentrations, between control and polluted samples were observed (PO0.
Also, number and kind of pollen flavonoid changes especially increasing flavonoid contents were observed in polluted plants comparing to control.
Studying seed and pollen characters can be used as air quality bioindicators.
Keywords: Air pollution, bioindicator.
Brassicaceae, flavonoids, pollen, power plant
INTRODUCTION
Thermal power plants are designed in a variety of fashions depending on the available fuel, mostly natural gas and heavy fuel oil.
Depending on the fired fuel, emission of thermal power plants is sulphuric and nitric Sulfur oxides constitute the most common pollution when burning heavy fuel oil (Noori et al.
By the reason Brassicaceae taxa importance from ecological, pastoral, medicinal and edible points, this study was done on 10 collected Brassicaceae taxa from the area and their Using plants for biomonitoring of air and soil quality may turn out to be successful, as they are simple, cheap and They can supplement the classical physicochemical By these types of techniques, the information on the pollutants can be derived from the study of the biological responses of plants to pollution (Malayeri et al.
Many authors studied on reactions of plants to air pollutants, and those species suitable as plant bioindicators (Gottardini et al.
Bosac et al.
investigated the impact of O3 and SO2 on reproductive development of oil seed rape (Brassica napu.
, pollen germination.
Pollen as a biologic allergen is a sensitive bioindicator of atmospheric pollution and provides particularly original and interesting information on the potential adverse effects of pollutants on living organisms (Yosefi et al.
Air pollution affects on growth, development, morphobiometrical and phytochemical pollen grains parameters.
Many pollutants have direct impact on the pollen physiology.
and indirect impact on its ontogenesis via their effects on plant It may be pointed out that this ontogenesis is also subordinated to the other environmental factors .
tmospheric and/or edaphi.
acting on the producing Of course, pollen does not indicate levels of pollutants, but it measures their biological impact (Gottardini et al.
Pollen viability and morphology were used for fluoride pollution biomonitoring (Malayeri et al.
Studies on collected pollen from polluted and unpolluted areas have shown contradictory results.
Acid rain could affect developmental process of pollen grains.
Exine was not formed in some case of plants that treated by different acid solutions (Chehregany et al.
Pollination with SO2 fumigated pollen resulted in reduced seed production and weight (Elke 1.
Low-level O3 and/or SO2 exposure causes a linear decline in soybean yield and high-level exposure caused significant linear reductions of 4-7% in mass per seed, seed number and weight (Sprugel et al.
Reich and Amundson 1.
Studies on extracts of exposed Quercus rubra.
Festuca elatior and Ulmas pumila pollen grains with CO.
SO2 and NO2 using twodimensional thin layer chromatography and SDS-gel electrophoresis (PAGE) methods showed changing amino NOORI et al.
Ae Brassicaceae characters and air pollution acids and molecular weight profiles.
Also, antigenic changes were observed in contaminated pollen comparing to control using double immune-diffusion method (Ruffin et al.
Air pollution effects on structures, proteins, and allergenicity of pollen grains using SDS-PAGE method showed decreasing protein content in all of polluted samples in response to air pollution.
(Majd et al.
Changing pollen protein profiles and appearing some new bands in DEP .
iesel exhaust particle.
-exposed pollen grains were observed in polluted Lilium martagon pollen extract comparing to control (Chehregany and Kouhkan 2.
Anthocyanins were as an indicator for plant responses to environmental stress (Chalker-Scot 1.
Polyphenolics were introduced as indicator to different environmental It shows plant metabolic adaptation with increasing flavonoids content responding to environmental pollution (Rezanezhad and Nasibi 2.
Rezanejad .
stated that phenolic compounds function as stress indicators because they accumulate to high levels in many plant tissues in response to a wide range of biotic and abiotic signals.
These compounds are involved in pollen development, pollination, pollen germination, and pollen tube growth (Rezanejad 2.
Feucht and Treutter .
studied the role of flavan-3-ols in plant defense.
They found that flavan-3-ols and other phenolic compounds as unsaturated oil acids protective are reported against free Also flavanols increase cell membrane protective agents (Feucht and Treutter 1.
Analyzing polluted pollen grain extracts, using HPLC showed increasing flavonoids level in polluted pollen in comparison with control (Rezanejad 2.
Such studies indicated that seeds and pollen grains provide essential information on biological impact of pollutants and they are good candidates for biomonitoring the atmospheric and edaphic In this study effects of Shazand Thermal Power Plant (Ira.
pollution on seeds characters, pollen fertility and flavonoids profile in 10 collected Brassicaceae taxa from the Power Plant area .
aving SO2.
CO.
NO) in comparison with control 40 km away from the power plant .
o this pollutan.
were investigated.
Also about using these characters as air pollution bioindicators is discussed.
MATERIALS AND METHODS
Collection of plant material, determination, and seed morpho-biometry Plant material of ten wild and native Brassicaceae taxa (Alyssum linifolium var.
Alyssum longistylum.
Alyssum marginatum.
Choriospora persica.
Clypeola lappacea.
Conringia perfoliata.
Descurainia sophia.
Goldbachia laevigata.
Isatis kotschyana and Neslia apiculat.
were collected .
ive samples of each tax.
from Shazand Thermal Power Plant .
ttp://w.
com/en/products/powe.
area located at NE of Shazand.
Markazi Province.
Iran and its geographical coordinates are 33A 53' 25" North, 49A 9' 43" East (Figure .
ttp://w.
com/iran/markazi/shazan.
Control samples were collected from 40 km distances of the Power Plant.
Herbarium vouchers were prepared.
Collected plant identified using available authoritative references (Rechinger Mobayen 1979.
Ghahreman 1976-2.
(Figure .
Seed morphobiometrical studies were done using a zoom binocular light microscope.
Seed length maximum (SLM), seed width maximum (SWM), seed length max/seed width max (SLM/SWM), seed diameter max (SDM), normal seed% (NS%) and abnormal seed% (AS%) measured and calculated for 10 plants of each taxon.
Round and having kernel seeds were considered normal versus wrinkled, holed and lacked kernel seeds as abnormal.
Figure 1.
Showing the Shazand Thermal Power Plant area located at NE of Shazand.
Markazi Province.
Iran N U S A N T A R A B I O S C I E N C E 10 .
: 96-104.
May 2018 Figure 2.
Showing habit of 10 studied Brassicaceae taxa.
Alyssum linifolium var.
Alyssum longstylum.
Alyssum marginatum.
Choriospora persica.
Clypeola lappacea.
Conringia perfoliata.
Descurainia sophia.
Goldbachia laevigata.
Isatis kotschyana.
Neslia apiculata Preparation and light microscopy of pollen grains Flowers of all collected control and polluted samples were fixed in Carnoy's solution .
fixative composed of glacial acetic acid and ethanol 3:.
Pollen grains were studied using Muntezing's acetocarmine method (Noori Five samples from each species were randomly chosen, and their anthers smashed on a microscopic slide.
Empty anther shells were removed, and one drop of the staining solution was applied to the pollen grains on the respective slide.
After uniformly dispersing the pollen and stains, they were covered with a cover slip.
24-48 hours later, the slides were observed by light microscopy (Galen i.
Leica.
Wetzlar.
German.
Palynological characters such as abnormality and sterility percentages were NOORI et al.
Ae Brassicaceae characters and air pollution determined for each control and polluted sample.
Pollens were categorized into two groups those with round exine and regular shape .
and these with wrinkled, shrunk, fragile exine and irregular shape .
Percentages of stained pollen .
and non stained pollen .
were also scored.
Preparation and extraction of pollen grains Pollen grain samples from mature fresh flower anthers were air dried after dissection of the flowers for flavonoids detection and identification.
For a comparative analysis of the flavonoids, small extracts of all the accessions were prepared by boiling 200 mg of powdered pollen grain for 2 min in 5 mL 70 % ethanol/water.
The mixture was cooled and left to extract for 24 h.
The extract was filtered, evaporated to dryness by rotary evaporation at 40CC and taken up in 2 mL 80 % MeOH/water for analysis by twodimensional paper chromatography .
-D PC) (Noori 2.
Flavonoid analysis by 2-D PC Aliquots of 10 times 2 AL of each extract were applied to the corner of a quarter sheet of chromatography paper (Whatman No .
The chromatograms were developed in the first direction with n-butanol-acetic acid-water=4:1:5 v/v, upper layer (BAW) and in the second direction with 15% aqueous acetic acid, with rutin (= quercetin 3-Orutinosid.
as standard.
After development, the chromatograms were viewed in UV 366 nm and any dark absorbing and fluorescent spots were marked.
Rf -values were calculated (Noori et al.
Methods of identification of the flavonoids After obtaining sufficient amounts of purified flavonoids, as in the case of the flavonoids from 10 Brassicaceae taxa pollen, they were identified by means of UV spectroscopy using shift reagents to investigate the substitution patterns of the flavonoids (Mabry et al.
Markham 1.
and by acid hydrolysis to identify the aglycone and sugar moieties.
Co-chromatography with standards was also performed where possible.
Flavonoid standards available for comparison during the study were apigenin, chrysin, genistein, hesperidin, isorhamnetin, kaempferol, luteolin, morin, myricetin, naringenin, quercetin, rhamnetin, rutin, tricine and vitexin, rutin (ILDIS 1.
from Merck.
Darmstadt.
Germany.
and luteolin from Sigma.
St.
Louis.
MO.
USA.
and the others from Fluka-Sigma-Aldrich.
Deisenhofen.
German.
Acid hydrolysis and identification of flavonoid aglycones After dissolving a small amount of each purified flavonoid .
5 m.
5 ml of 80% MeOH in a test tube, 2 ml of 2M HCl were added and the mixture was Then 2 ml of EtOAc were added and thoroughly mixed with the aqueous layer using a Whirley mixer after The upper EtOAc layer was removed and evaporated to dryness, dissolved in 0.
5 ml of MeOH and applied as spots on thin layer chromatograms .
The TLC plates were run in three solvents alongside standards to identify the aglycone moiety (Harborne 1.
Then TLC chromatograms were viewed in UV 254 nm and each spot Rf -values and color comparing to standards helped to kind of flavonoids identification.
Data analysis Seed morphobiometrical data results were analyzed by EXCEL.
Pollen abnormality and sterility percentages were Pollen grain flavonoids data were scored.
Then all of obtained data were analyzed by ANOVA.
Duncan, and Tukey tests (PO0.
using SPSS.
RESULTS AND DISCUSSION
Results Seed morphobiometrical studies results showed seed dimensions, and seed normality percentage had reduction in all of studied taxa except Neslia apiculata species that abnormal seed percentage was higher than in control comparing to polluted (Table .
Table 2 shows palynological data of polluted samples comparing to control using Muntezing's acetocarmine method.
Results showed all of studied control plant pollen grains were elliptic and tricolporate (Figure .
All of control samples of ten collected Brassicaceae taxa from 10 km distances of the Shazand Thermal Power Plant area exceptional G.
laevigata and I.
kotschyana showed 100% fertility in their pollen grains.
While, abnormality was observed in all of studied polluted samples.
Also their total sterility% Polluted I.
kotschyana had the most abnormal pollen grains .
8%) and maximum sterility .
5%).
Minimum abnormality pollen grains .
9%) and minimum sterility .
92%) were observed in polluted G.
Pollen grain flavonoids data of ten studied Brassicaceae taxa from the Shazand Thermal Power Plant area.
Iran comparing to control using 2-DPC and TLC methods have been shown in Table 3.
As the Table shows flavonoid sulfates and flavon C-and C- /O-glucosides were observed in all of studied control and polluted samples.
Aglycones were not found in control and polluted samples of Alyssum longstylum.
laevigata and N.
apiculata species while both control and polluted samples of A.
marginatum and C.
lappacea species lacked.
Polluted samples of C.
sophia and C.
perfoliata had aglycones comparing to control but I.
kotschyana and A.
linifolium var.
taxa control samples had aglycones in comparison with This study revealed high level changing in kind and number of pollen grain flavonoids between control and polluted samples.
Flavonoids number increased in polluted samples comparing to control with the exception of A.
marginatum and G.
laevigata species.
Polluted N.
and A.
marginatum showed the most increasing total flavonoids and C.
lappacea was at least.
Fourteen flavonoids compounds consisting of apigenin, chrysin, genistein, hesperidin, isorhamnetin, kaempferol, luteolin, morphine, myricetin, naringenin, quercetin, rhamnetin, rutin, tricine, and vitexin were found in control and polluted samples of studied taxa.
Table 4 shows statistical analysis results of morphobiometrical, palynological and flavonoids studied data using SPSS by Tukey and Duncan N U S A N T A R A B I O S C I E N C E 10 .
: 96-104.
May 2018 Table 1.
Seed morphobiometrical data of 10 collected Brassicaceae taxa from the Shazand (Ara.
Thermal Power Plant area.
Iran comparing to control SLM .
SWM .
MASD Taxa SDM .
NS% SLM/SWM AS% C P C P Alyssum linifolium var.
6A0.
5A0.
97A0.
97A0.
22A0.
2A0.
54 92 84 8 16
Alyssum longistylum 14A0.
05A0.
51A0.
61A0.
66A0.
56A0.
27 97 94 3
Alyssum marginatum 26A0.
8A0.
56A0.
3A0.
54A0.
5A0.
38 99 98 1
Choriospora persica 61A0.
56A0.
22A0.
25A0.
75A0.
74A0.
14 94 81 6 19
Clypeola lappacea 05A0.
67A0.
77A0.
47A0.
58A0.
46A0.
13 94 67 6 33
Conringia perfoliata 23A0.
88A0.
13A0.
80A0.
94A0.
46A0.
35 97 90 3 10
Descurainia sophia 03A0.
02A0.
52A0.
54A0.
40A0.
37A0.
88 95 89 5 11
Goldbachia laevigata 60 A0.
57 A0.
58A0.
56A0.
30A0.
30A0.
65 80 75 20 25
Isatis kotschyana 50A0.
50A0.
94A0.
63A0.
79A0.
40A0.
76 87 74 13 26
Neslia apiculata 76A0.
60A0.
27A0.
12A0.
06A0.
87A0.
43 93 99 7
Note: C=control.
P=polluted.
SLM= seed length max.
SWM= seed width max.
SLM/SWM=seed length max/seed width max.
SDM =
seed diameter max.
NS%= normal seed%.
AS%= abnormal seed%.
Table 2.
Palynological data of polluted samples of 10 collected Brassicaceae from the Shazand Thermal Power Plant area.
Iran comparing to control using Muntezing's acetocarmine method Taxa Normal pollen grain Abnormal pollen grain Normality Fertility Sterility Abnormality Fertility Sterility C P Alyssum linifolium var.
Alyssum longistylum Alyssum marginatum Choriospora persica Clypeola lappacea Conringia perfoliata Descurainia sophia Goldbachia laevigata Isatis kotschyana Neslia apiculata Note: C=control.
P=polluted Total fertility Total sterility 50 100 91.
9 100 100 0
8 100 100 0
4 100 100 0
4 100 100 0
1 100 99.
2 100 93.
2 25 92.
6 60 32.
Neslia Isatis Goldbachia Descurainia Conringia Clypeola Choriospora Alyssum Alyssum Taxa Alyssum linifolium var.
Table 3.
Pollen grain flavonoids data of ten studied Brassicaceae taxa from the Shazand Thermal Power Plant area.
Iran comparing to control using 2-DPC and TLC methods Characters C P C P
P C P C P C
P C P
C P C
Total flavonoids number
7 9 5
8 11 10 11 9 12 6
8 11 7
9 11 7
Flavonoid sulphates number
4 6 3
Flavon C-and C- /O-glucosides 2
3 3 2
Aglycones 0 0 1
Apigenin
- - - Chrysin
Genistein
- -
Isorhamnetin
- A
Kaempferol
Luteolin
Morin
- - Myricetin
- - - Naringenin
- - - A
Quercetin
Rhamnetin
A - A
Rutin
- - - Tricin
A - Vitexin
- - A
- Note: Scored characters: - .
on flavonoid=.
A (UV absorbance < 0.
1, rare concentration of flavonoid=.
, .
ew concentration of flavonoid=.
, .
iddle concentration of flavonoid=.
, .
igh concentration of flavonoid=.
NOORI et al.
Ae Brassicaceae characters and air pollution Also Table 5 shows selected significant parameters by ANOVA (PO0.
Table 4.
Statistical analysis results of seed and pollen morphobiometrical and pollen flavonoids data using SPSS by Tukey and Duncan tests (PO0.
Discussion As Table 1 shows pollution have affected on seed dimensions in all of studied polluted samples.
Seed dimensions have reduced in polluted plants comparing to But seed width has been exceptionally increased in Alyssum longstylum and Descurainia sophia.
All of polluted sample fruits had more abnormal seeds number .
ithout kernel, holed or wrinkle.
in comparison with control (Figure .
laevigata and N.
apiculata polluted fruits had less abnormal seeds than control.
Statistical analysis results in Tukey and Duncan tests showed that power plant pollutant had been caused normal seed number reduction in all of species exceptional G.
laevigata and N.
apiculata that confirmed Agrawal and Deepak .
SO2
effects on soybean caused seed number reduction and consequently crop decreasing.
Sprugel et al.
showed 79 g/Lit SO2 on soybean significantly caused seed weight and number reduction that mainly reduced crop (Sprugel et al.
Reich and Amundson 1.
Using Muntezing's acetocarmine pollen grain staining method revealed decreasing of fertility in all of ten studied polluted Brassicaceae taxa (Table .
The method is one of the most widely used staining techniques for pollen viability estimation and shows sharp differences in smearing advanced pollen grains from abnormal ones.
As a result, sharply stained pollen grains were considered as potential of fertile and viable ones, and partially stained, no stained, or weakly stained pollen grains were considered nonviable and sterile (Malayeri et al.
As Table 2 shows the highest abnormality and sterility percentages pollen grain was observed in I.
kotschyana in comparison with the same species control samples and also other polluted species.
The results show stimulation and inhibition of these pollen characteristics depend on the plant species as well as on the pollutant and its Finally, this study indicates that air pollution can induce several abnormality and sterility in plant pollen Test of homogeneity of Levene Apigenin Chrysin Genistin Isorhamnetin Luteolin Morin Myricetin Naringenin Rhamnetin Rutin Tricin Vitexin Flavonoid Sulphates Number Aglycones Number Flavonoid C-and C-/O.
glucosides Number Total Flavonoid Number Apigenin concentration Chrysin concentration Genistin concentration Isorhamnetin concentration Kaempferol concentration Luteolin concentration Morin concentration Myricetin concentration Naringenin concentration Quercetin concentration Rhamnetin concentration Rutin concentration Tricin concentration Vitexin concentration Seed length max.
Seed width max.
Seed length max/Seed width max.
Seed diameter max.
Abnormal seed percentage Abnormal pollen percentage Sterile pollen percentage Note: *Bold numbers significant (PO0.
DF2
SIG.
Table 5.
Selected seed and pollen morphobiometrical and pollen flavonoids significant parameters by ANOVA (PO0.
ANOVA
Kaempferol concentration Between Groups Within Groups Total Abnormal seed % Between Groups Within Groups Total Abnrormal pollen grain% Between Groups Within Groups Total Sterile pollen% Between Groups Within Groups Total Note: *Bold numbers significant (PO0.
Sum of Squares Mean square Sig N U S A N T A R A B I O S C I E N C E 10 .
: 96-104.
May 2018 Figure 3.
Showing fruits of 10 studied Brassicaceae taxa.
Alyssum linifolium var.
Alyssum longstylum.
Alyssum marginatum.
Choriospora persica.
Clypeola lappacea.
Conringia perfoliata.
Descurainia sophia.
Goldbachia laevigata.
Isatis kotschyana.
Neslia apiculata.
Strip = 1 mm NOORI et al.
Ae Brassicaceae characters and air pollution Phytochemical results in Table 3 show many changes in kind and number of flavonoids in polluted plants compared to control.
The total flavonoids increasing were observed in polluted samples of N.
apiculata and A.
Flavonoids number increase in polluted Robinia pseudoacacia grown under fluoride pollutant (Noori et al.
Also using HPLC method on polluted pollen grain ethanolic extracts of Lagerstroemia speciosa.
Spartium junceum.
Petunia hybrid and Thuja orientalis showed that air pollution caused high-level flavonoids accumulation in polluted samples in comparison with control (Rezanezhad Flavonoid sulfates number in A.
kotschyana and N.
apiculata polluted pollen grains were more than control while these flavonoids in A.
marginatum and G.
were less than and A.
linifolium and C.
lappacea did not show any changes.
As our TLC results show, some flavonoids appeared in polluted samples while disappearance others were observed.
Quercetin and kaempferol were found in all of control and polluted samples while rhamnetin lacked.
Tricin was just found in polluted A.
longistylum sample.
Chrysin found in all of control and polluted samples except A.
Presence of morin and myricetin in polluted C.
and I.
kotschyana and absence in their control show creation or deletion of flavonoids under pollutant effects.
Studies on eight angiosperm plants collected from Shazand Oil Refinery area.
Iran using two-dimensional paper and thin layer chromatography showed flavonoids kind and number changing in polluted plants comparing to control.
Also, kaempferol appearance in polluted Amaranthus chlorostachys samples comparing to control was observed (Kamalabadi 2.
Robles et al.
studied phenols and flavonoids in Aleppo pine needles as bioindicators of air pollution.
Their results showed that total flavonoids are useful bioindicators for ozone pollution .
ignificant positive correlations between total flavonols and ozone pollutio.
Sulfur dioxide pollution is distinguished by low concentrations in quercetin, isorhamnetin, and kaempferol .
ignificant negative correlations between these simple flavonols and the concentrations of SO.
) The work confirms strong interest of using the phenolic compounds of Pinus halepensis as biological indicators of air quality (Robles et Rubin et al.
found induction of isoflavonoid production in Phaseolus vulgaris leaves by ozone, sulfur dioxide and herbicide stress (Rubin et al.
Total flavonoids reduction was observed in treated plants with heat and SO2 (Lee et al.
Lavola, 1.
Increasing total flavonoids and phenolic compounds were observed in collected Catharanthus roseus and Ocimum sanctum from NO2 and SO2 polluted area.
These plants can be used as bioindicators (Qayoom et al, 2.
Increasing or reduction of flavonoids under air pollutant can show pollution effects on flavonoids.
Phytochemical changes in aluminum smelter industry area vegetation were observed with appearance or disappearance of flavonoids in fluoride polluted plants (Noori et al.
Statistical analysis results of morphobiometrical and flavonoids data using SPSS by Tukey and Duncan tests in Table 4 show morin existence and morin and kaempferol concentrations, abnormal seed and pollen percentages and sterile pollen% are significant (PO0.
Also.
Table 5 shows selected seed phytochemicals significant parameters by ANOVA method (PO0.
and confirms that kaempferol concentration, abnormal seed and pollen grains percentages and sterile pollen% are three significant parameters.
These results have coincided on Sprugel et al.
about seeds.
Noori et al.
about pollen and Kamalabadi .
about It is known that plant flavonoids pattern depends on genetics factors and ecological conditions.
Results of the study confirmed that pollen grains provide essential information on biological impact of pollutants and they are good candidates for biomonitoring the atmospheric and edaphic pollutions.
As Ruffin et al.
studied effects of some environmental gaseous pollutants on pollen-wall proteins of certain airborne pollen They used pure pollen grains of Ulmus pumila.
Quercus rubra.
Pinus taeda, and Festuca elatior were used as a biological screen to test the effects of certain commonly occurring atmospheric pollutants on pollen wall protein (Ruffin et al.
Also.
Rezanejad .
studied air pollution effects on structure, proteins, and flavonoids in pollen grains of Thuja orientalis.
Pollutions such as power plant pollutant are caused by different environmental effects.
The most pollutants of power plant are SO2.
NO.
NO2.
CO, and CO2 that affected on ecosystem and plant vegetation.
These atmospheric pollutants affect pollen morphobiometry, fertility, and Their effective on plants caused different morphological, physiological and phytochemical responses to pollutants that reduced pollen and seed production and their viability and fertility.
Changes in flavonoids kind and number especially flavonoids sulfates production or increasing in polluted plant in comparison with control plant defensive reactions to pollutants.
It seems that environmental pollution reduced plant resistance against catching disease and pest attack and increased plant Then studying pollen characters such as viability, morphobiometery, their phenols and flavonoids compounds and also seed production and characters can be used as air quality bioindicators.
ACKNOWLEDGEMENTS
The authors would like to thank of Mrs.
and Mr.
Faraki for their help in collecting of our samples.
REFERENCES