International Research Journal of Environment Sciences________________________________ ISSN 2319–1414Vol. 3(3), 31-38, March (2014) Int. Res. J. Environment Sci. International Science Congress Association 31 Study of Morphological Characteristics of Spinach Irrigated with Industrial waste water of Bhiwadi, Rajasthan, IndiaSaini Madhvi, Sharma K.C. and Sharma ManjuDepartment of Botany, University of Rajasthan, Jaipur, INDIAAvailable online at: www.isca.in, www.isca.me Received 19th February 2014, revised 25th February 2014, accepted 21st March 2014 AbstractIn the present investigation industrial area of Bhiwadi, (Alwar district) was selected as study area. The plants of spinach were treated with 3 types of water smaples i. ground water ii. Treated water (CETP outlet) and iii. untreated water (CETP inlet) and analyzed for morphological characters . Results showed that waste water increased stem length, root length ,fresh and dry weight, leaf length and leaf breadth and decreased seedling length, vigour index, and germination percentage of seeds. Waste water effects leaf colour and shape and seed colour, seed size and spermoderm patterns. Results reveated that waste water irrigation brought up negative changes in most of the important growth parameters of plants and hence this water is not found suitable for crop irrigation. Keywords: Industrial waste water, spinach, morphological characterstics, spermoderm patterns. Introduction Industrialization is rapidly increasing to fulfill human needs. Waste water from industries is released in water bodies. The untreated and partially treated waste water contains fertilizers, pesticides, textiles, pharmaceuticals and various types of other chemicals which have been proven harmful to living beings particularly animals, plants and human beings1-4. Physio-chemical parameters of water are induced due to discharge of untreated or partially treated industrial waste and sewage waste into water bodies5-7. The untreated or partially treated waste water from industries are continuously used in irrigating the agricultural fields in developing countries including India8-10. Continous use of this waste water for irrigation effects soil quality11,12. The industrial effluents, in lower quantities provide macro and micro nutrient requirements of the plants. But higher quantitiy becomes harmful. Leafy green vegetables like spinach, kale, chards and others are the most potent superfood. Leafy green vegetables are rich source of minerals like calcium, magnesium, iron and potassium, vitamin K and C. they are low in fat, high in protein, and high in dietary fiber and also contain a lot of water. Leafy vegetables like spinach are more sensitive to toxic pollutants than other vegetable crops because of higher amounts of accumulation of heavy metals in their leafy growth and long term intake of these leafy vegetables cause clinical problems both to animals and human beings. The present study was aimed at analyzing quality of water from industrial area of Bhiwadi, and its effect on morphological characteristics of spinach. Material and Methods Study area: Bhiwadi is called gateway of Rajasthan. It is located in Tijara tehsil of Alwar Distt.. It comes under Delhi NCR region. It is 55 km away from Indira Gandhi International airport, New delhi and 200 km from Jaipur. It is a industrial hub. It has 1500 small and large industries including MNC industrial units and steel, furnance, engineering, pharmaceuticals, textiles etc. Collection of water samples: The study was conducted with effluents released from industries at Bhiwari, Alwar. There are industries like steel, electronics, textiles, drugs,chemicals food processing etc. The waste water released from these industries were collected at the main outlet point where combined effluent from the industries is being disposed and inlet point of CETP (common effuleunt treatment plant)/untreated water and other water samples were collected from CETP outlet point (treated water) and third water sample is ground water. Experimental set up and raising of plants: 3 plots of 6.5 × 4.5 size were prepared and genetically uniform seeds of spinach were sown in each plot. Uniform irrigation schedule was followed at all the sites to maintain similar moisture condition throughout the growth of plants. Names of the 3 plots were given as GW, inlet, outlet plots. When the plants were growing morphological characterstics (vegetative growth) like stem length, root length, petiole length, leaf colour, fresh and dry weight, seed germination percentage, seed colour, seed size, and seed surface patterns were recorded. International Research Journal of Environment Sciences______________________________________________ ISSN 2319–1414 Vol. 3(3), 31-38, March (2014) Int. Res. J. Environment Sci. International Science Congress Association 32 INDIA RAJASTHAN ALWAR BHIWADI (STUDY AREA) Figure-1 Bhiwadi (study area) map For plant sampling, five replicates from each sub plot were collected randomly from ground water, treated water and untreated water irrigated plots. Seed germination percentage, seedling length, root and shoot length, petiole length, leaf length and breadth and seed size were quantified. leaf shape, leaf colour, seed colour and were also observed. Seed surface patterns were studied by SEM .For this seeds were affixed on aluminium stub with help of transparent adhesive. Seeds coated with gold and examined at a range of magnification in a EVO 18 Scanning Electron Microscope at USIC department, University of Rajasthan, Jaipur. For fresh weight estimation, plant samples were separately washed to remove the soil particles and weighed. For estimation of dry weight plant sample were dried in shade. Plant samples were weighed separately and biomass was expressed in gm. For germination percentage, 50 sterilized seeds of spinach were placed in sterilized petri dishes containing filter paper. They were kept wet continuously by adding different water samples. The number of germinated seeds in each Petri dish was recorded daily up to 10 days and expressed as % seed germination. After 10 days vigor index (VI) was calculated13. VI = Germination% × Seedling Growth Results and Discussion Leaf length, leaf breadth and petiole length, leaf area are 2.96±.343, 2.27±.194, 3.74±.380 cm respectively; 1.54±.194, 0.74±.258, 1.81±.293 cm respectively and 2.26±.509,1.73±.245, 2.93 ±.330 cm respectively, 126.3±13.05,158.85±25.65, 202.2±32.8 cm as observed in plants grown in ground water, treated water and untreated water respectively (figure 2). Irrigation with sewage has increased in sorghum leaf width and also, increased the yield rate14. Seedling length was 6.52±.852,5.62±.825 ,5.7±2.05 cm as observed in ground water, treated water and untreated water respectively figure-3. Shoot and seedling length was reduced after metal treatments15-16. This could be because of reduction in meristematic cells present in the region and some cotyledonary and endosperm enzymes which became active and began to digest the stored food. This resulted in not proper supply of food to the radical and plumule. The value for vigour index is 521.6, 359.68 and 319.2 cm respectively and germination percentage is 80%, 64%, and 56% was observed in plants grown in ground water, treated water and untreated water samples respectively. Figure-4, 7. Growth characteristics such as plant health (i.e. radicle length and plumule lengths) and seed vigour index increased with 50% effluent irrigation and decreased with 100% (undiluted) effluent irrigation at all soil column heights17-18. Fresh weight of leaves is 0.3565±..031, 0.383±.018 and 0.6745±.014 gm of plants grown in ground water, treated water and untreated water respectively. Dry weight of leaves is 0.0256±.0081, 0.0233±.0063, and 0.0886±.0074 gm of plants grown in ground water, treated water, untreated water respectively figure-2. Effects of different percentages of effluent irrigation with wastewater on corn yield 19. Results showed, the highest grain yield and total biomass were observed in 100% urban treatment and with reduction of wastewater, the yield decreased too20. Cotton yield irrigated with wastewater was more than that in irrigation with typical water 21. Stem length is the highest of plants irrigated with treated water 1.79 ±.871 cm, followed by height of plants irrigated with treated water 1.63±.560cm and minimum height 1.38 ±.565cm of plants grown in ground water figure-6. Root length is the highest of plants irrigated with treated water 19.1±0.77 cm, followed by height of plants irrigated with treated water 17.4±0.258cm and minimum height 11±0.840 cm figure-5. International Research Journal of Environment Sciences______________________________________________ ISSN 2319–1414 Vol. 3(3), 31-38, March (2014) Int. Res. J. Environment Sci. International Science Congress Association 33 Table–1 Showing comparison of morphological characters in plants irrigated with ground water, treated water and untreated water industrial waste water S No. charcterstics Ground water Treated (outlet) Untreated (inlet) 1 Stem length 1.38 ±0.871 1.79 ±0.560 1.63 ±0.565 2 Root length 11±0.840 cm 17.4 ±0.757 cm 19.1 ± 0.770cm 3 Leaf length 2.96 ± 0.343 cm 2.27 ±0.194 cm 3.74 ± 0.380 cm 4 Leaf breadth 1.54 ±0.194 cm 0.74 ±0.258 cm 1.81 ±0.293 cm 5 Petiole length 2.26 ±0.509 cm 1.73 ± 0.245 cm 2.93 ±0.330 cm 6 Leaf shape ovate ovate Curling in leaf margin 7 Leaf colour green Green +yellowing Green with brown spot 8 Seed size 3.7*2.0 mm 3.6*2.0mm 3.8*2.1mm 9 Germination % 80% 64% 56% 10 Seedling length 6.52±0.852 5.62±0.825 5.7±2.05 12 Vigour index 521.6 cm 359.68 cm 319.2 cm 13 Fresh weight 0.3565 ±0.031gm 0.383 ±.018 gm 0.6745 ±0.04gm 14 Dry weight 0.02566 ±.0081 gm 0.0233±0.0063 gm 0.08866 ±0.0074 gm 15 Leaf area 126.3±13.05 cm 2 158.85±25.65 cm 2 202.2±32.08 cm 2 Shoot length showed a significant higher value in waste water irrigated plants as compared to the controls. This in support of the earlier studies in Hardwic kia binata and in Wheat22-23. Irrigation with sewage water caused increasing tailoring, stem length, root length, panicle length and number of spikes in rice24. And similar results weret observed in barley in the Boyerahmad region of Iran25. However there was increase in certain parameters like root - stem length, length of leaf and fresh and dry weight etc. in the present investigation also. But this is due to industrial waste water which generally contains heavy metals. And literature reveals that micronutrients found in waste water may be beneficial for plant growth but several micronutrients which are heavy metals may produce undesirable effects on plants at higher concentration26. And this is supported by present data which indicated that visible symptoms of toxicity appeared in untreated plants, such as brown spots on leaf and chlorosis and ovate shape with curling in leaf margin. Colour of leaf as green with yellow and ovate shape is observed in plants irrigated with treated waters. These symptomps were not obeserved in plants grown in ground water figure-11,12. The retardation in growth and development of plants could be due to high uptake of heavy metals and their accumulation in plants27-28. Seed colour is greyish light brown in plants grown in ground water, brownish black coloured and black coloured in plants grown in of treated water and untreated water respectively Figure-9. Seed size are 3.7 ×2mm, 3.6 ×2mm, 3.5 ×2.0 mm respectively of plants growing in ground water , treated water and untreated water which is non significant. However the seed surface patterns as revealed by Scanning electron microscope showed warty, net like and prickly surface with flaty elevated muri of thin strands in seeds from plant irrigated with ground water. The flaty elevated muri become regular and much condensed in treated water palnts and densly populated with irregular arrangements and condensed populations in plants grown in untreated waste water. Figure-2 Fresh and dry weight of plants grown in ground water, treated water (outlet) and untreated water (inlet) Figure-3 Seedling length of plants grown in ground water, treated water (outlet) and untreated water (inlet) International Research Journal of Environment Sciences______________________________________________ ISSN 2319–1414 Vol. 3(3), 31-38, March (2014) Int. Res. J. Environment Sci. International Science Congress Association 34 Figure-4 Germination percentage of plants grown in ground water, treated water (outlet) and untreated water (inlet) Figure-5 Root length percentage of plants grown in ground water, treated water (outlet) and untreated water (inlet) Figure-6 Stem length, leaf length, leaf breadth, petiole length of plants grown in ground water, treated water (outlet) and untreated water (inlet) Figure-7 Vigour index of plants grown in ground water, treated water (outlet) and untreated water (inlet) Figure-8 Leaf area of palnts grown in ground water, treated water (outlet) and untreated water (inlet) Conclusion The data presented here seem to reveal that industrial waste water used for crop irrigation significantly affected quality parameters of spinach as compared to ground water irrigation. Seedling growth, germination percentage and other morphological characters may be considered as an indicator of metal stress on plant vigour. International Research Journal of Environment Sciences______________________________________________ ISSN 2319–1414 Vol. 3(3), 31-38, March (2014) Int. Res. J. Environment Sci. International Science Congress Association 35 Figure-9 Showing seed germination in ground water, treated and untreated water figure A-C and seeds obtained after harvesting from the experimental palnts irrigated respectively ground water treated and untreated water D-F International Research Journal of Environment Sciences______________________________________________ ISSN 2319–1414 Vol. 3(3), 31-38, March (2014) Int. Res. J. Environment Sci. International Science Congress Association 36 Figure-10 Showing plants grown in ground water figure A-B and plants grown in treated water C-D Figure-11 Showing plants grown in untreated water International Research Journal of Environment Sciences______________________________________________ ISSN 2319–1414 Vol. 3(3), 31-38, March (2014) Int. Res. J. Environment Sci. 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