_??_ 1987 by Cytologia, Tokyo C ytologia 52: , Meiotic Abnormalities Induced by Dye Industry Waste Water in Chlorophytum amaniense Engler

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1 _??_ 1987 by Cytologia, Tokyo C ytologia 52: , 1987 Meiotic Abnormalities Induced by Dye Industry Waste Water in Chlorophytum amaniense Engler Accepted July, 28, 1986 R. K. Somashekar Department of Botany, Bangalore University, Bangalore-56056, India Biocides and other chemicals are known to induce chromosomal abnormalities both in meiotic and mitotic cells of plants (Amar and Ali 1968, 1969, Amar and Farah 1968, Ahmed and Grant 1972, Mohan 1975, Soliman and Al-Najja 1980 and Somashekar et al. 1983, 1984). Waste waters of industrial origin contain many organic and inorganic chemicals and represent a complex chemical unit. So far studies made have shown that they waste waters also induce a variety of chromosomal aberrations in the mitotic cells (Ravindran and Ravindran 1978, Sh anthamurthy and Rangaswamy 1979 Somashekar and Arekal 1983, Somashekar and Gowda 1983, Somashekar et al. 1985). The present paper describes the meiotic abnormalities in Chtorophytum amaniense induced by dye industry waste water special emphasis on chromosomal abnormalities induced. Materials and methods The industry is located 4km away from Mysore city, Karnataka, India and it generates 50,000 Its of waste water per day. The waste water is discharged into the nearby sewage canal situated 1-1km. from the factory. Standard methods (APHA 1975) were employed during the collection, preservation and analysis of the waste water. A Perkin Elmer Model 403 Atomic absorption spectrophotometer was used for the estimation of heavy metals. Ample number of young flower buds of Chtorophytum amaniense were dipped in 25, 50, 75 and 100 per cent effluents for 3, 6 and 9h. The accessory parts of the inflorescence were remov ed before dipping, to ensure proper treatment. Control plants were also maintained. Under each treatment as many as 6-8 plants were considered and the experiment was repeated. Flower buds gathered after treatment were fixed in Carnoy's fluid and stained using acetocarmine smear method. Abnormalities were counted at meta-, ana- and telophases. Results The average values of physico-chemical characters of the effluents are given in Table 1 (100%). The effluents were acidic and characteristic due to the presence of 2% Azored, 1-2% Acid Violet and Triphenyl Methane. They also contained heavy metals such as chromium, copper and zinc in considerable amounts. Treatment of Chtorophytum amaniense (2n=14) plants waste water produced a singi fi cant percentage of abnormal PMCs per plant (Table 2). The mean percentage of abnormal PMCs ranged from 3.66 }0.19 at 25% and 3h treatment period to at 100% concentration and 9h treatment. Mostly the percentage abnormalities induced remained dose dependent. In comparison the control, significantly high percentage of abnormal cells were observed in the treated series. A total of 2,000 PMCs were observed to estimate the percentage of abnormal PMCs at various stages of division (Table 3). Comparatively many abnormal PMCs were observed at metaphase II, an average percent at anaphase I and the lowest at telophase II. As many as 5.12, 8.13 and 16.87% of abnormal PMCs were observed

2 648 R. K. Somashekar Cytologia 52 at metaphase I, against 6.03, 9.14 and at metaphase II, a treatment period ranging from 3 to 9h. Nearly 7.91 and 6.34% of abnormal PMCs were observed at anaphase I and II during 9h and the respective values during telophase I and II were 5.67 and Treatment of flower buds effluents produced a variety of abnormalities. At 3h period, cells "undefined abnormalities" were noticed and an increase in the duration of treatment the abnormalities became distinct (Table 4). The cells laggards (Figs. D, F, K) and stickiness of chromosomes (Figs. C, H, I, J, K), tripolar and multipolar cells (Fig. E) Table 1. Physico-chemical characteristics of effluents (100% concentration), chemical characteristics expressed in mg/l Table 2. Percentage of abnormal PMCs per Chlorophytum amaniense plant after treatment effluents were encountered frequently. Fragmentation (Fig. G), bridges (Fig. H) and micronuclei (Fig. I) were observed less frequently and precocious movement was the other deviation (Fig. L). Discussion The foregoing observations indicate that stickiness of the chromosomes is the common effect produced after longer durations of treatment and the formation of univalents is a feature

3 1987 Meiotic Abnormalities Induced by Dye Industry Waste Water in Chlorophytum Figs. A-L. ~ 680. C, ~ 630. broken E, A and B, condensed chromosomes tripolar fused chromosomes irregularly. ~660. anaphase. ~695. fragments. ~565. telophase metaphase and scattered a micronuclei. ~430. laggard. ~480. H, F, metaphase anaphase L, irregularly D, laggard. ~580. bridge. ~520. J, chromosome metaphase distributed anaphase bundle at metaphase. laggard. G, I, 649 anaphase chromosomes at metaphase. ~465. precocious movement. ~360. at K,

4 Table 3. Percentage of abnormal PMCs observed after treatment effluents

5 1987 Meiotic Abnormalities Induced by Dye Industry Waste Water in Chlorophytum 651 at shorter duration. Pesticides are also known to produce a similar effect (Amar and Farah 1976, 1984). The disturbed cells at meta and anaphase II showed irregularly scattered chromosomes in the cells. The occurrence of less percentage of abnormal cells during Al and TI is attributed either to the recovery of cells or to the probability that, such PMCs do not complete the meiotic cycle as observed by Amar and Farah (1968). Multipolar cells were frequent at TII whereas lagging occurred frequently at Al or out fragments. Laggards failed to reach the pole before uncoiling of chromosomes to form daughter nuclei and they either disintegrated or formed micronuclei. The bridges appear to form by the failure of terminalization in biva lents, especially from the chromosomes stretched towards poles. It is also probable that bridges result by stickiness of chromosome ends. The breaks might have been produced by the direct effect on the structure of chromosome. The occurrence of univalents and an increase in their frequency in MI seems to be the outcome of disturbances during pairing of homologous chro mosomes. Univalents appear to arise from partial or complete lack of homologous chromo Table 4. Percentage of abnormalities observed in C. amaniense after treatment effluents some pairing, and the whole process regarded as desynapsis and asynapsis has been attributed to gene mutation and chromosome breakage (Riley and Law 1965). The micronuclei further result in polyads and are responsible for the production of sterile pollen (Patil and Bora 1961), and others such as non-oriented chromosome at MII and Al as well as laggards are known to produce non-viable gametes. The cells breaks undergo reciprocal translocation, resulting in the formation of multivalents. The stickiness disturbes the identity of the chromosome and therefore, is responsible for the organization of non-viable gametes. Summary Treatment of flower buds of Chlorophytum amaniense different concentrations of dye industry waste water resulted in the production of high percentage of abonormal PMCs. The abnormalities observed included stickiness of chromosomes, laggards, bridges, fragments and micronulcei. The higher frequency of stickiness observed is apparently responsible for some

6 652 R. K. Somashekar Cytologia 52 of the above abnormalities. It is concluded that dye industry waste water rich amounts of heavy metals act as potential mutagens. References Ahmed, M. and Grant, W. F Cytological effects of some mercurial fungicides Panagon-15 on Tradescantia and Vicia faba root tips. Mutation Res. 14: Amer, S. M. and Ali, E. M Cytological effects of pesticides II. Meiotic effects of some phenols. Cytologia 33: and Cytological effects of pesticides IV. Mitotic effects of some phenols. Cytologia 34: and Farah, O. R Cytological effects of pesticides III. Meiotic effects of N-methyl-l-naphthyl car bamate 'Sevin'. Cytologia 33: and Cytological effects of pesticides VIII. Effects of carbamate pesticides "IPC", "Rogar" and "Duphar" on Vicia faba, Cytologia 41: and Cytological effects of pesticides XIII. Meiotic effects of the insecticide "Duraban". Cy tologia 49: APHA, AWWA and WPCF Standard methods for the examination of water, waste water and sewage. 14th Edn. APHA. Inc. New York. Mohan, S. T Cytological effects of Pesticide plantvax and Vitavax on somatic cells of Allium cepa Curr. Sci. 44 (22): Patil, S. H. and Bora, K. C Meiotic abnormalities induced by X ray in Arachis hypogea. Indian J. Genet. Plant Breed. 21: Ravindran, J. N. and Ravindran. S Cytological irregularities induced by water polluted factory efflu ents: A preliminary report. Cytologia 43: Riley, R. and Law C. N Genetic variations in chromosome pairing. Advance Genetics 13: Shanthamurthy, K. B. and Rangaswamy, V Cytological effects of paper mills effluents on somatic cells of Allium cepa. Cytologia 44: Soliman, S. A. and Al-Najja, R. N Cytological effects of fungicides II. Chromosomal aberrations in duced by Vitavax-200 and Dithane S-60 in meiotic cells of wheat and two related species. Cytologia 45: Somashekar, R. K., Shetty, G. H. P. and Gowda, M. T. G Mitodepressive and clastogenic effects of Peperidine. Mutation Res. 118: and Arekal, G. D Chromosomal aberrations induced by electroplating waste water. Cytologia 48: and Gowda, M. T. G Cellular damage induced by food processing industry waste waters in Allium cepa. Curr. Sci. 52: , - and Venkatasubbaiah, P Cytological effects of fungicide Topsin in Allium cepa. Cytologia 49: , Gurudev, M. R. and Siddaramaiah Somatic cell abnormalities induced by dye manufacturing industry water. Cytologia 50: