62 CURRENT ADVANCES IN AGRICULTURAL SCIENCES 4(1): JUNE 2012 Current Advances in Agricultural Sciences 4(1): 62-63 (June 2012) ISSN 0975-2315 SHORT COMMUNICATION Impact of sugar factory effluent on chlorophyll and protein contents of Cicer arietinum and Tigonella foenum-gracecum ANUBHA SHARMA, AMIT KUMAR* and TS DHAKA Department of Botany, D.A.V. College, Muzaffarnagar-251 001 (Uttar Pradesh), India *Email of corresponding author: amit_bio80@yahoo.com Received: 11 August 2011; Revised accepted: 04 April 2012 Key words: Chlorophyll, Cicer arietinum, Protein, Sugar factory effluent, Trigonella foenum-graecum The effluent application to various crops may be an ideal alternative method for recycling industrial waste. The sugar factory effluent contains many inorganic and organic substances, many of which may be phytotoxic at higher concentration, but useful for the growth and development of the plants. Chlorophyll is the pigment of photosynthesis process may be taken as a parameter of photosynthetic capacity of the plants. Quantitative analysis of the protein also gives an idea about the total nitrogen content of the plants. Therefore, during the present study these two biochemical parameters were studied and analyzed quantitatively. Cicer arietinum and Trigonella foenum-graecum crops are selected to assess their sensitivity to various concentrations of sugar factory effluent, as these crops are important and widely used in Western Uttar Pradesh. The pure sugar factory effluent is diluted with the double distilled water to get different concentration, viz. 5, 10, 25, 50 and 75%; 100% (effluent without any dilution) and 0% (distilled water served as control). For estimation of chlorophyll and proteins the seedling were harvested after five week (35 Days) of sowing. Plant tissues weighed and analyzed for these parameters. Chlorophyll content was determined spectrophotometrically as per the procedure of Arnon (1949). Fresh plant leaves ground in 80% acetone with little CaCO 3 in mortar and pestle. The paste was centrifuged at a speed of 4000 rpm for five minutes. Supernatant was taken and 80% acetone is added to make the volume of 50 ml. The absorbance observed at 663 and 645 nm on a systonics spectrophotometer. Carotenoid content estimated in 100 mg of fresh leaves homogenized in 80% acetone with a pinch of sodium bicarbonate. The homogenate centrifuged at 3000 rpm for ten minutes and volume made up 20 ml and kept in dark for 24 hours. Absorbance was recorded at 470 and 510 nm. Estimation of protein contents was done as per methods described by Bradford (1976). All samples were harvested at 14 th day after sowing on ice mixture (NaCl + Ice). After washing with prechilled double distilled water, samples was blotted with filter paper and then weighed. Samples were crushed in prechilled mortar-pestle on ice mixture in cold Tris buffer (containing 50 mM Tris pH 8.5, 10 mM EDTA (sodium salt) pH 8.2, 10 mM MgCl2, 20 mM KCL, 5 mM â-mecroploethanol. The slurry was centrifuged at 5°C at 10000 rpm for about 20 minutes to get the clear supernatant. The volume of supernatant was recorded. Egg albumin was used for the preparation of standard calibration curve. Briefly, diluted dye reagent (5 ml) was added to the 0.1 ml aliquots of sample of standards (20, 40 µg ml -1 of protein). Buffer (0.1 ml) was taken as blank. After a period of 5 minutes to 1 hour, OD 595 versus reagent blank was measured and standard calibration curve was prepared. Samples OD 595 was also measured. The concentrations were read from standard curve and calculated mg g -1 fresh weight of shoot. Chlorophyll and carotenoid content increased at lower concentration of effluent i.e. 5 and 10% as compared to control values in both the crops (Table 1). Further increase in effluent concentrations decreased these photosynthetic parameters. Maximum inhibition to chlorophyll content is observed with 75% sugar factory effluent in both the crops. However, no data recorded for 100% sugar factory effluent treatment, as there is no seed germination at this concentration. Besides, total chlorophyll contents, chlorophyll a and chlorophyll b are also recorded in the same sample. The values of chlorophyll a and chlorophyll b also showed the same trend as total chlorophyll influenced by different concentration of effluent. Protein content in tested seedlings showed similar trend as chlorophyll being maximum at 10% effluent concentration above this concentration started protein synthesis inhibition.