Effect of Heavy Metals Like Mercury (Hg++), Cadmium (Cd++) and Lead (Pb++) on the Spore Germination of Pterisvittatal; the Common Road Side Fern
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International Journal of Science, Technology & Management www.ijstm.com Volume No 04, Special Issue No. 01, March 2015 ISSN (online): 2394-1537 EFFECT OF HEAVY METALS LIKE MERCURY (HG++), CADMIUM (CD++) AND LEAD (PB++) ON THE SPORE GERMINATION OF PTERISVITTATAL; THE COMMON ROAD SIDE FERN Dr. Mausumi Biswas1, Dr. Pushanjali Khare2, Dr. Namita Kumari 3 1, 2, 3 Department of Botany, Magadh Mahila College, Patna University, Patna, (India) ABSTRACT Study the effect of different heavy metal salts like PbCl2, HgCl2 andCdCl2 on the germination time and germination percentage. As PbCl2, CdCl2 and HgCl2 are common pollutants present in the automobile exhaust and industrial effluents and this particular cryptogam is common in roadside so the effects of these salts are studied by culturing the spores in culture medium in various concentrations. Keywords: Germination Percentage, Germination Time, Pteris Vittata L., Pollutants, Cdcl2, Pbcl2 and Cdcl2. I. INTRODUCTION The ferns are one of the most beautiful cryptogams on the earth. They found all over the world, from high mountains to sea level. Among them, PterisVittata L is the common road side fern found in all places. They are present in all environmental conditions and are also show their sensitivity to the heavy metal salts. Heavy metal salts are present in the environment as pollutants. Generally heavy metal salts like HgCl2, CdCl2 and PbCl2 are present in the air or water as the pollutant. Effect of these salts shows antagonistic and lethal effect on the germination of the spores. , and can be correlated with the germination percentage and germination time. In all the big cities there are several industries like lead smelting industry, Chlore alkali industry and Battery industry are present. These industries are alarmingly discharging Pb++, Hg++, Cd++, Sn++ andZn++ as pollutants in the atmosphere. This situation adds support to the report that normally 25 – 30% mercury burden of these atmospheres is man – made (See Das, Sharma and Talukdar; 1982). Micronucleus formation in quite common is Crepis capillaris when its seeds are treated with 0.1 and 0.01 ppm/lit cadmium salt. Inorganic salts of Mercury in different concentrations also have strong effects on flowering plants. Dutta and Sen (1992) reported the injurious effect caused by these pollutants on ferns. Gupta, Devi and Singh (1992) reported the injurious effects caused by these pollutants on ferns. Gupta, Devi and Singh (1992) reported the effect of cadmium on the entire life cycle of gametophytes of Ceratopteris thalictroides. The effect of spores germination and gametophyte development in some ferns has been studied by Gupta and Devi (1992) . Ho and Tai (1985) recorded the effect of pollutants on the ultra structures of Pteris vittata. Petersen , Arnold , Lynch and Price (1980) and also Francis and Petersen (1986) recorded the effect of these heavy metals ( Lead , Mercury , Cadmium etc) on the percent spore germination of Onoclea sensibilis . 194 | P a g e International Journal of Science, Technology & Management www.ijstm.com Volume No 04, Special Issue No. 01, March 2015 ISSN (online): 2394-1537 II. MATERIAL AND METHOD Mature spores of Pteris vittata L. a homosporus road side fern, were collected. For studying spore morphology, both acetolysed and non acetolysed spore preparations were used. Acetolysis was done following the method of Erdtman(1952).Spore measurements represent the average of ten readings. For synchronous culture freshly collected spores were sown. Prior to sowing, the spores were sterilized. For sterilization, spores were treated with sterile distilled water. These were then transferred to 5% calcium hypochlorite for 3 – 5 minutes, centrifuged, washed twice with sterilized distilled water and again centrifuged. Finally the aqueous suspension of the spores was taken in a 10 ml sterilized pipette for inoculation. The cultures were maintained at temperature of 240C + 20C under 16 hours of white fluorescent illumination in 24 hours cycle at 300 ft. c. of light intensity in the BOD. Germinating spores were distinguished from the non germinating ones by staining with chloral hydrate – acetocarmine following method of Edward and Miller (1972). Germinating spores following such treatment appeared pink, white the non germinating ones turned dark brown. Moore’s medium supplemented with Nitsch’s trace element (Kato 1969) was used in liquid and also in solid state. This medium has been used as the basal medium for the study of germinating spores. Medium is autoclaved for 15 minute at 15 lb/ inch2 pressure and the PH was adjusted to 5.6. ++ ++ ++ To study the effect of heavy metal salts of Pb , Hg and Cd , the salts lead chloride (PbCl2 ), Cadmium chloride ( CdCl2) and mercuric chloride (HgCl2 ) are added in different concentration of 0.001ppm/lit ; 0.01 ppm/lit; 0.1ppm/lit and 1ppm/lit in the basal medium . The four sets of cultures are compared with the basal Moore’s medium which is treated as control. Germination percentage, Germination time of spores are studied in different intervals and tabulated in the table for the discussion. III. OBSERVATION Germination time and germination percentage of the spores of Pteris vittata is studied in the basal medium and also in the media supplemented with salts of HgCl2 , CdCl2 and PbCl2 in 1ppm/lit , 0.1 ppm/lit ; 0.01 ppm/lit and 0.001 ppm/lit . It is found that spores of Pteris vittata L. germinate 84 hours after the inoculation , which is considered as the standard time for the germination . In control germination percentage is 82.8% which is the mean of 10 readings. It is found that 1ppm/lit concentration of PbCl2 , CdCl2 and HgCl2 become lethal for the spores and no germination takes place.The percentage of germination decreases with the increase concentration of heavy metal salts . In 0.1ppm/lit HgCl2 germination percentage is 11.6% and germination time is 126 hours . In 0.01 ppm/lit HgCl2, germination percentage is 22.8% and germination time is 99 hours; and in 0.001 ppm/lit HgCl2 germination percentage is 42.9% and germination time is 89 hours. In 0.1 ppm/lit PbCl2, germination percentage is 15.2% and germination time is 115 hours; in 0.01 ppm/lit PbCl2 germination percentage is 35.5% and germination time is 92 hours and in 0.001 ppm/lit germination percentage is 51.6% and germination time is 82 hours. 195 | P a g e International Journal of Science, Technology & Management www.ijstm.com Volume No 04, Special Issue No. 01, March 2015 ISSN (online): 2394-1537 In 0.1 ppm/lit CdCl2 , germination percentage is 13.7% and germination time is 120 hours ; in 0.01 ppm/lit germination percentage is 23.8% and germination time is 95 hours , where as in 0.001 ppm/lit CdCl2 germination percentage is 41.2% and germination time is 87 hours . Table 1.1:- Germination Percentage and Germination Time Of Pterisvittata in Control and In Different Concentrations of Salts of Three Heavy Metals Control Germination percentage Germination time ( in hours ) 82.8 84 Heavy metals salts Concentration of heavy Germination Germination time metals percentage (in hours ) 1 ppm/lit No germination No germination HgCI2 0.1 ppm/lit 11.6 126 0.01 ppm/lit 22.8 99 0.001 ppm/lit 42.9 89 1 ppm/lit No germination No germination PbCI2 0.1 ppm/lit 15.2 115 0.01 ppm/lit 35.5 92 0.001 ppm/lit 51.6 82 1 ppm/lit No germination No germination CdCI2 0.2 ppm/lit 13.7 120 0.02 ppm/lit 23.8 95 0.001 ppm/lit 41.2 87 IV. DISCUSSION It has been found that germination percentage is 82.8% in the control and germination time is 74 hours after inoculation .All the heavy metal salts HgCI2 , CdCI2 and PbCI2 show total lethal effect on the germination of the spore when they are treated with 1 ppm/lit concentration . It has been found that 0.1 ppm/lit HgCI2 shows most drastic effect on the spore germination percentage which is 11.6%. Germination time also increases in 0.1 ppm/lit HgCI2 which is 126 hours after inoculation; where as it is 115 hours in PbCI2 and 120 hours in CdCI2 salt concentration. Only 22.8% spores germinate in 0.01 ppm/lit of HgCI2 where as 35.5% h spores germinate in 0.01 ppm/lit PbCI2 and 23.8% spore germination 0.01 ppm/lit CdCl2 salt concentration . Germination time is 99 hours in 0.01 ppm/lit HgCl2 ; 92 hours in 0.01 ppm/lit PbCl2 and 95 hours in 0.01 ppm/lit CdCl2 concentration ; 51.6% in 0.001 ppm/lit PbCl2 ; and 41.2% in 0.001 ppm/lit CdCl2. Germination time is 89 hours in 0.001 ppm/lit HgCl2 ; 82 hours in 0.001 ppm/lit PbCl2 ; 87 hours in 0.001 ppm/lit CdCl2 . It has been clear from the above result that HgCl2 salts shows most harmful effect on the germination of the spores of Pteris vittata . It has been also observed that germination of spores Pteris vittata delayed in the presence of different heavy metals (Bondada, B.R and Lena Qiying M.A; 2003). Cadmium and lead shows antagonistic effect on spore germination of Pteris vittata (Gupta and Devi , 2005 ) Kamachi at al ( 2005 ) have observed that spore germination in Pteridium , Lygodium japonicaum and Pteris vittata shows adverse effect in the presence of heavy metal salt like Cd++ and Pb++ . It has been found that Pteris vittata which is a common road side fern 196 | P a g e International Journal of Science, Technology & Management www.ijstm.com Volume No 04, Special Issue No. 01, March 2015 ISSN (online): 2394-1537 shows delayed spore germination and abnormal growth collected from different polluted area containing heavy metal pollutants ( Khare.