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EFFECT OF THERMAL POWER PLANT WASTEWATER AND COAL FLY ASH ON ROOT NODULATION, GROWTH, YIELD AND QUALITY OF CICER ARIETINUM L.

ABSTRACT

THESIS SUBMITTED FOR THE AWARD OF THE DEGREE OF Bettor of $I)ilogopl)p IN BOTANY

IRFAN AHMAD ^^

DEPARTMENT OF BOTANY ALIGARH MUSLIM UNIVERSITY ALIGARH (INDIA) 2004 '•*' J-™.*^*^ EFFECT OF THERMAL POWER PLANT WASTEWATER

AND COAL FLY ASH ON ROOT NODULATION, GROWTH,

YIELD AND QUALITY OF CICER ARIETINUM L.

Irfan Ahmad

Abstract of the thesis submitted to Aligarh Muslim University,

Aligarh, India for the Degree of Doctor of Philosophy in Botany, 2004.

Five pot experiments were carried out based on factorial

randomized block design during the rabi seasons of 1999-2002 with the

aim to study the utility of thermal power plant wastewater (TPPW) and

fly ash (FA) along with nitrogen and phosphorus doses on two cultivars

of chickpea. The data was mostly significant. Various parameters of

, fly ash and were also analysed for physico-chemical

properties.

Experiments I and II were performed together during the rabi

season of 1999-2000, to study the comparative effect of TPPW and GW

and four levels of fly ash (FAQ, FAio, FA20, FA40) on the basis of growth, yield and quality parameters of chickpea cultivars BG-256 and

Avarodhi. TPPW proved beneficial except for seed protein content which remained unaffected. Among the various levels of fly ash, FAio proved optimum in comparison to FA20 and FA40. The data was also pooled to compare the two cultivars and it was BG-256 which performed

better than Avarodhi.

Experiments III and IV were also conducted simultaneously in the

year 2000-2001 on the same cultivars. TPPW again proved better for

most of the parameters studied including NRA, nodulation and seed

yield confirming the findings of the first two experiments. However, it

may be noted that in these two trials based on nitrogen doses, protein

content was enhanced due to the waste water application. Treatment Nio

proved optimum, whereas N20 was at luxury consumption and N30 was

toxic. The data of the two experiments was also pooled and it was again

BG-256 which performed better than Avarodhi.

Experiment V was conducted during the rabi season of 2001-2002

under five levels of phosphorus (Po, Pio, P20, P30, P40) supplemented with

a uniform starter dose of 10 kg ha"' fly ash, 10 kg N ha'' and 20 kg K ha"' on cultivar BG-256. TPPW proved effective and P30 proved the optimum dose for most of the parameters including NRA, nodulation, seed yield and protein content, while P40 was excessive and Pio, P20 were deficient. Nodulation and protein content increased with increasing levels of phosphorus. EFFECT OF THERMAL POWER PLANT WASTEWATER AND COAL FLY ASH ON ROOT NODULATION, GROWTH, YIELD AND QUALITY OF CICER ARIETINUM L.

^^ THESIS SUBMITTED FOR THE AWARD OF THE DEGREE OF Bottor of pi)tlogopt)p IN BOTANY , \ t...<

IRFAN AHMAD

DEPARTMENT OF BOTANY ALIGARH MUSLIM UNIVERSITY ALIGARH (INDIA) *** 2004 „,'S*ri'.>.

16919 'dedicated (To (iJVly Barents Ariflnam Department of Botany M.Sc, Ph.D. Aligarh Muslim University Professor Aligarh - 202002. [email protected]

Dated: l-o • \i^- c^ 1

CERTIFICATE

This is to certify that the thesis entitled "Effect of Thermal Power Plant

Wastewater and Coal Fly Ash on Root Nodulation, Growth, Yield and

Quality of Cicer arietinum L." submitted in partial fulfilment of the requirement for the degree of Doctor of Philosophy in Botany is a faithful record of the bonafide research work carried out at Aligarh Muslim

University, Aligarh by Mr. Irfan Ahmad under my guidance and supervision and that no part of it has been submitted for any other degree or diploma.

Supervisor of re'sCarch ACKNOWLEDGEAIENT I mth use of this pmious ojiporijiinitj to CXJJKSS my hctirtfdt ^mtituk and slnccrcst thmh to my Icumei teacher aid sufiervisor Professor Arif mam for his aUe ^ikiia, mtho]d which this work would not have materialised. I owe sincere thanks to the Head of the vhint vhysiolo^ Division, Dej)artment of 'Botany, Ali^arh Muslim university, Ali^rh and EX-chalrman Professor samiulkh for j^roviMn/j necessary Infrastmcture, facilities during the tenure of my research work, i owe a sense of^atitnk to Professor M.M.R.K. Af'idifor his adviu, encouragement and heljifd criticism during this investigation. I am eXtrmdy thankful to Professor ishrat Husain Khan, chairman, Deyar\ment of 'Botany, Ali^arh Muslim university, Ali^arh for the encouragement and helji that he rendered at almost every stejf during ike course of my research work. I eitend my thanks and dcejf sense of gratitude to my learned teachers Professor A. Ahmad, Dr. F. Mohammad, Dr. N.A. Khan, Dr. M.M.A. Khan, Dej)artment of 'Botany, Ali^arh Muslim university, Ali^arhfor their constant helj) and^idaue. \nfad, tkey had ken a constant source of inspiration and encouragement for me. m this hectic world we live in, we very rarely take time to comjilement yeoyle for a jol well done. But I really want to say a special thanh to my seniors Dr. s. Hay at. Dr. M. Mokn, Dr. A. Hussain, Dr. Q^Fariduddin, Dr. Z.M. Azam, Dr. R. Mir, Dr. s. ]avi^. Dr. F.A. sheMi, Dr. A. Afelit/ir, Ms. s. Alvi as they cause hayfmess wherever they^o, hit others whenever they^o. I am highly thankfd to my coka^es shahk, Manzer, Pervez, Faisal, Kashif Naeem, Sarljeet, Azam, saha and Dilshada as they have provided me with the fill strength of their help and support. Coha^es are not just friends hut more than that, "when nights are long and friends are few, l sit alone and think of you with a lovely heart and silent tears all I wish is you to k here kcmse you are the full support which none can provide, they provide happiness like a hllle hath." 1 am also extremely thani^l to my friends shiv, Afeel, Preeti and Ahidfor their constant support as they are like the pillars on our porch. Sometimes they hold us up and smdimcs tfiey kdn on us. somdimcs its just enough to ho\i that thj arc stmdln^ hj. Without tkm i couU not think ofmj research worfe to le comjileted in time. I especidly wish to thmk to mj friend shouht 'Bhai who hdsjioured immense low on me and have ken a source of sujfjiort md encouragement during the course of my research worii. He is just like the mirrors of the hleidoscojie who will always shine in the hottom of my heart and don't let others know, if the day comes when ifly, and^o uji in the sky, as I am so far, I'll write your name on every star, so you look uy and see, how much you really mean to me. I am highly fateful to my yarents and all family memkrs especially my elder hrother. Jawed hhai, for jfrovidin^ me indispensable support and continuous encouragement showered with lots of love and prayers. I am also thanl^l to the staff menders mzish Ihai, Masood Ihai, Kehana apa, javed Ihai, shahid Wai, shakir hhai and Hayat hhai for constant help, i want to commend on having as a staff of very competent, efficient and courteous employees. Moreover, I am abo eitremely thankful to the gardeners who provided ^ood help to me. I am also ^ratefd to Mis Ihai for extending his adept services on computer for typing the nwiuscript. With all faith in the almighty i place this work in the hands of my eUminer with the hope that he will kar with the shortcoming that mi^ht have crept into this thesis inadvertently.

(irfan Ahmad} CONTENTS

INTRODUCTION 1

REVIEW OF LITERATURE 4

MATERIALS AND METHODS 37

RESULTS 58

DISCUSSION 103

SUMMARY 118

BIBLIOGRAPHY 120

APPENDIX i introduction Chapter-1 INTRODUCTION Water and plant nutrients, among others inputs, are the two most important factors for normal growth of any crop. The former, being the most abundant molecular species in plants, is also a carrier of nutrients. By definition, a nutrient is that which nourishes and out of which the substances of plants are synthesized. In this cense, therefore, water also performs a nutritional role. It is noteworthy that covering about 1404 million cubic kilometers its quantity on earth, is enormous. However, 97.6% of it is in the oceans and unsuitable for irrigation due to its high content, while about 2.0% is tied up in glaciers, ice caps and snow fields. Thus, only about 0.4% fresh water is available for plants, animals and human beings (Cunningham and Saigo, 1995). Since India is a monsoon-dependent land and the bulk of rainfall is confined to a brief period only, a large part of the country remains deficient in water supply for a greater part of the year. The situation here, like some other countries (China, Kenya, Nigeria), is much worse than that of the more fortunate ones. Our per capita renewable fresh water supply in 1999, for instance, was less than 2,000 m^ annum'' which was far less than that of Canada and the USA and even Australia, Brazil and Malaysia (Anonymous, 2001). This is because fresh water resources worldwide are unequally distributed. For example, Asia with 60% of the world population has only 36% of river runoff whereas. South America, with just 6% of the population, enjoys about 26% runoff Similarly, whereas about 60% of the available water consumed each year worldwide is diverted for irrigation, for Asia, which has about two third of the wodd's irrigated land, the figure is as high as 85%o due to unscientific irrigation. A situation therefore arises when most Indian farmers have no option but to grow their crops totally under rain fed conditions. A few also make indiscriminate and unscientific use of municipal or industrial wastewater, including thermal power plant wastewater (TPPW), which is easily available due to growing urbanization and industrialization. One such source of water in India is based on 89 coal fired thermal power plants of 100 MW or higher capacity, generating about 66,860 MW electricity Introduction

day"'. These burn about 65% of the total coal produced in India and simultaneously discharge huge quantities of water as well as fly ash (FA). Both are rich in a variety of beneficial as well as toxic elements, including N, P, K, Ca, Mg, S04^ Cr, B, Mo, Se and Sr (Klien et al, 1975; Asano et a/., 1985). It is noteworthy that efforts made to modify the properties of low grade agricultural soil by adding FA have proved encouraging in improving soil fertility and crop yield. Thus, it has been observed that FA not only improves the water holding capacity of some sandy (Salter and Williams, 1967; Chang et al., 1989) but also augments their capacity for providing some essential nutrients (Hill and Lamp, 1980). Admittedly, being phytotoxic as well as deficient, specially in N, the utility of FA in agriculture is limited. This could, however, be enhanced by blending it with sludge or by irrigating the crop with water containing this essential nutrient. Interestingly, plants that are Nj fixers can tolerate comparatively high salinity and B toxicity and, therefore, are successful colonizers under these poor soil conditions. Such limited utilization of FA alone for a limited number of crops including N2 fixing legumes, is known. So is that of TPPW. It also helps at least partly, in solving the problem of their disposal. In addition, it also decreases the total dependence of our farmers upon chemical whose indiscriminate use in the last century from the sixties onward due to Government of India sponsored subsidies and attraction of higher productivity for short term economic gains has created serious problem of nutrient pollution. In view of the experience gained in some advanced countries, plant nutrition experts at various Indian centres, including Aligarh, are trying to shift the emphasis on maximizing crop production from over application of chemical fertilizers to minimum nutrient emission to safeguard not only the already deteriorated farm soil but also the nearby water bodies. In this context, it may be pointed out that pulses (grain legumes) can fix about 50% of their total nitrogen requirements with rates of fixation going upto 100 kg ha'' (Pepper, 2000). They have been playing a vital role in sustainable eco-friendly agriculture. Pulses, being an integral part of vegetarian diet in the Indian sub-continent, are a known rich source of protein. However, it must Ic admitted that the area Introduction

under their cultivation has not increased in proportion to population explosion. Consequently the per capita availability of pulses has progressively declined from 60.7g day"' in 1951 to nearly 36g in 2000 against the FAO/WHO recommendation of 80g (Sud, 2001). Chickpea (Cicer arietinum L.), an important pulse crop grown throughout the country, accounts for more than a third of the area under pulses and about 40% of their production in India, the average annual area and production being about 7-8 million hectares and about 4-5 millions tonnes respectively (Anonymous, 2002). In view of the known beneficial role of wastewater from various sources and of FA separately in augmenting crop productivity and for helping the disposal management of the two waste products and keeping the importance of pulses in mind, as explained above, the present author proposed a hypothesis that chickpea, a popular Nj fixing pulse crop, could be grown profitably using minimal inputs by judiciously substituting them with the two waste products of TPP, namely fly ash (FA) and waste water (TPPW). To test the hypothesis, it was decided to conduct five pot experiments during the years 1999-2002 with the aim : 1. To compare the effect of TPPW and ground water (GW) on chickpea {Cicer arietinum L.) cv. BG-256 (Experiment I) and cv. Avarodhi (Experiment II) under four levels of FA application. 2. To compare the effect of TPPW and GW on chickpea cv. BG-256 (Experiment 111) and cv. Avarodhi (Experiment IV) under four levels of basal N along with the best dose of FA obtained in Experiments I & II. 3. To test the comparative efficacy of TPPW and GW for the cultivation of the better performing cultivar of chickpea (Experiment V) under five levels of basal P along with the best dose of FA and N obtained in the previously conducted Experiments. J^erature CONTENTS

REVIEW OF LITERATURE

Pase no. •*& 2.1 Effect of industrial wastewater on crop plants 4

2.2 Effect of fly ash on crop plants 19

2.3 Effect of NPK on chickpea (Cicer arietinum L.) 31 Chapter-2 REVIEW OF LITERATURE

2.1 Industrial wastewater on crop plants Rapid industrialization in India, during the last few decades, in the form of chemical, dairy, pharmaceutical, mining, distillery, , paper and pulp, petrochemical, textile, , machinery, vehicle, food processing, tannery factories in addition to coal fired thermal power plants, has burdened the land and resulted in deterioration of air, soil as well as water. However, some of the liquid discharges resulting from these manufacturing and industrial processes are not necessarily harmful, since it has been observed that after proper treatment or dilution, such wastewaters are beneficial at least for agricultural purpose. Thus, judicious use of wastewater may be one of the feasible options to prevent wastewater from being an environmental hazard. In addition, it has been found in number of trials that it can serve as a potential source of some essential and beneficial nutrients. In fact, the utilization of treated wastewater as liquid fertilizer makes sense because (i) it replaces and conserves mineral fertilizer that would otherwise be introduced into the environment (ii) the presence of organic material improves soil structure and (iii) an alternative disposal method could be environmentally damaging, expensive or limited by space. Literature pertaining to the studies on the industrial wastewater in relation to leguminous as well as non-leguminous crops has been reviewed in the following pages. Srivastava and Sahai (1987) at Gorakhpur performed petriplate as well as pot experiments to investigate the effect of distillery effluents on Cicer ahetinum (chickpea). 15 days old seedlings were picked, and the length of root and shoot was recorded. For the study of growth, earthen pots were taken and 250ml of each concentration diluted with tap water was given. In their study percentage of germination and seed germination index (SGI) decreased with increase in concentration and no germination took place in undiluted effluent. Although in 75%, some seeds germinated but the seedlings could not survive beyond 4 days. The maximum root length was recorded at 2.5% while root and shoot lengths and leaf area increased upto 5% but declined at higher concentrations. In addition to the biomass, Review of Literature

net primary productivity, chlorophyll content, reproductive capacity, seed output and seed protein also exhibited progressive increase upto 5%. The pods also appeared earlier in plants grown in 5%. They were of the opinion that the lower concentrations of distillery effluent could be beneficial and can thus be used as liquid fertilizer. Further studies were conducted by Sahai et al. (1985). They evaluated the same effluents on growth behaviour oiPhaseolus radiatus (black gram) and in 1987 Sahai and Neelam carried out another experiment to study the effect of distillery effluent mixed with fertilizer factory effluent on black gram and made more or less similar observations. Mukherjee and Sahai (1988) while taking Cajanus cajan (pigeon pea) further observed that 50% effluent provided optimum condition for germination, seed germination index, seed output and dry weight. Seedling establishment was 100% upto 50% effluent and shoot length was maximum at 2.5%. During the same year Sahai and Srivastava studied the effect of fertilizer factory effluents on Brassica oleracea Capitata (cabbage) and Brassica oleracea Botrytis (cauliflower). Decrease in germination and speed of germination index with increase in effluent concentration was reported by them, while Neelam and Sahai (1989) performed another study on Vigtm radiata (green gram). Respective lengths of root and shoot, plant biomass and N uptake were markedly increased when plants were grown with 10% distillery effluent. Total N in root, stem and leaf increased even upto 30%. Similarly distillery and sugar mill effluents were taken by Srivastava (1996) who reported that some crops showed moderate tolerance and better biomass when irrigated with diluted effluent. On the other hand concentrated effluent showed deleterious effect. Patra et al. (1989) observed the yield and economics of gram, Brassica compestris (mustard), Triticum aestivum (wheat), Pisiim sativum (pea) and cauliflower, in soils at Phulbani in Orissa. Crops were irrigated with various concentrations of paper mill sludge. At 0.5%, gram showed maximum response followed by cauliflower, peas and wheat while mustard showed better response under 0.25% concentration. Chaudhary et al. (1989) at Darbhanga also observed the impact of paper mill effluent on Hordeum vulgare (barley) in petriplates and noted gradual decrease in the germination percentage with increase in effluent concentration. This Review of Liteiatuie effluent was also tested by Misra and Behra (1990) at Berhampur during their study on Oryza sativa (rice) seedlings. Percent germination, water imbibing capacity, pigment, carbohydrate and protein content showed decreasing trend with increase in effluent concentration and time. Protein content was found to be most sensitive macrorr:olecule affected. Balaram et al. (2000) also at Berhampur studied the effect of paper mill, sugar factory and chloro alkali factory effluents and observed degradation of photosynthetic pigment in intact leaves. Similarly Srivastava (1991) at Jabalpur tested paper mill and chloro alkali effluent on Raphanus sativus (radish) and Alium cepa (onion) and reported that the latter was highly deleterious for germination and early grov^lh performance as compared to former. Agarwal and Chaturvedi (1995) at Faizabad, in a pot experiment, observed the effect of different concentrations of liquid waste discharged from a paper mill on wheat. Chlorophyll 'a', 'b' and total chlorophyll contents were considerably decreased, and the reduction in chlorophyll was more with increasing concentration and age of the plant. Baruah and Das (1997) performed a petriplate experiment on similar effluent at Guwahati. They reported that effluent polluted soil caused delay in seed germination and reduction in final germination percentage of rice by 12.5% compared with seeds in unpolluted soil. Also at Guwahati Dutta and Boissya (1997) noted that higher concentration inhibited the germination and growth of seedlings. Rice seed collected from effluent affected area were less viable and even the viable seeds showed delay in germination. Again Baruah and Das (1998) performed a petriplate experiment and noted delay, retardation and decline in germination and seedling growth under paper mill effluent. Similarly Dutta and Boissya (1999a, b) revealed that NPK contents of rice from paper mill effluent affected areas showed significant differences in growth and productivity. The effluents were further tested and it was observeu that chlorophyll content and leaf area could not be directly correlated with both affected and control rice plants. Dutta and Boissya (2000) also reported that the effluent significantly reduced each of the studied yield parameters, when compared to rice plant grown in areas beyond the reach of paper mill effluent. Dutta (1999) also tested paper mill effluent at Nagaon and reported that water that mixed with effluent was harmful to the growing paddy plants in general and injurious to the standing Review of Literature

paddy crop in particular. In a petriplate study by Sundaramoorthy and Kunjithapatham (2000) on six varieties of groundnut {Arachis hypogea) seeds were soaked in 0, 25, 50, 75 and 100% effluent. It was noted that germination percentage, seedling growth and dry weight decreased with increasing concentration. Ready and Borse (2001) at Pravarnagar also carried out a petriplate experiment on Trigonellafoenum-graecum L. (fenugreek) and reported that at 25% significant increase in germination percentage and other growth parameters was observed. Further increase in effluent concentration decreased the observed parameters. Agarwal and Gupta (1992) while investigating the effect of nitrogenous fertilizer factory effluent on seedling growth and biochemical properties of chickpea and mustard at Kota, reported that effluent showed deleterious effect. The inhibitory effect was more on radicle than hypocotyl and the effluent was responsible for significant decrease of pigment concentration in seedlings of their study. Jabeen and Saxena (1990) at Kanpur conducted petriplate as well as pot experiments on pea. 5% of distillery and 2.5% of fertilizer factory effluent proved favourable and thus exhibited an increase in dry matter, pigment content and protein content. In their opinion both the effluents could be favourably used for irrigation afler proper dilution. Goswami and Naik (1992) at Raipur, investigated the effect of fertilizer factory effluent on Cyamopsis tetragonoloha Taub. (cluster bean). Chlorophyll content improved under 10% effluent, but higher concentration adversely affected it. At the same place earlier Sharma ei al. (1990) performed a pot as well as field experiments to study the effect of steel plant wastewater on some crops. Wastewater was applied to Linum usitatissimum (linseed) in the field and to Sesamum indicum (sesame) and Phaseolus vulgaris (frenchbean) in pots. The plants showed decrease in Ca and Mg and increase in P concentrations. Fe was decreased in sesame and french bean but increased in linseed. They were of the opinion that the steel plant effluent should not be used for irrigation. Subramani et al. (1998) at Annamalai nagar investigated the impact of fertilizer factory effluent on cowpea {Vigna ungiculala L. Walp.). In their observation, 10% effluent was beneficial for overall growth. Earlier in 1995 Subramani et al. performed a petriplate experiment on green gram. Hyacinth Review of"Liteiatiiie

(Eichhoh:ia crassipes mort. Solms) plants were grown for 5 days in raw distillery effluent to get biologically treated effluent. Green gram seedlings were watered with various concentrations of raw as well as treated effluent. Seedlings at 5% and 10% concentration of biologically treated effluent showed increase in all growth parameters. Subramani et al. (1995) performed another petriplate experiment and reported decrease in germination, growth, yield and productivity of green gram with increasing distillery effluent concentration. In 1999, they further studied the growth behaviour of Ceratophyllum demersum L. (hornwort). Plants were grown for five days and reported that biologically treated effluent promoted the growth and yield. At the same place Sundaramoorthy et al. (200C) conducted a petriplate experiment to study the impact of fertilizer factory effluent on green gram, black gram, groundnut. Glycine max (soybean), paddy and Sorghum viilgare (sorghum). They reported beneficial effect at 10% effluent, beyond which seedling growth decreased. They conducted another petriplate experiment on groundnut varieties and reported that germination percentage and seedling growth increased with 1 to 10% effluent. The same crop was tested by Sundaramoorthy and Lakshmi (2000) under tannery effluent. Among the various varieties studied, TMV-4 showed better performance under tannery effluent and it proved to be tolerant while variety VRI-4 proved susceptible. Srivastava and Pandey (1999) at Faizabad, while considering the effect of fertilizer factory effluent, reported significant reduction in total chlorophyll content and bioniass of some aquatic mesophytes. At the same place earlier Khan and Srivastava (1996) conducted a field trial to assess the suitability of distillery effluent in agriculture. Field was filled with effluent prior to sowing. After 20 days field was tilled, crops were grown and subsequent irrigation was made with fresh water. After observing the resuhs, they recommended two to three diluted distillery effluent irrigations. The same source of effluent was also tested by Ahmad el al. (2000) at Rawalpindi (Pakistan) on wheat, barley, pea, spinach, fenugreek, Coriandum sativum (coriander), cabbage and Brassica rapa (turnip). They suggested remedial measures for the hazardous pollutants after observing the effect of effluent on these crops. At Shillong, Shukla and Moitra (1995) studied the clrect of 0, 25, 50, 75 and Review of Literature

100% effluent of integrated steel plant on growth parameters of chickpea, Phaseolus mungo (mungbean), maize and rice. They, after observing the resuhs, reported that lower concentrations were beneficial whereas higher sho^d deleterious effect on germination and seedling growth when seeds were soaked in various concentrations of effluent. Maize showed lowest tolerance to the effluent. Rubber factory effluent was tested by Sharma and Habib (1995) at Bareilly to study the bioaccumulation of Mg, Pb, Cr and Zn in two varieties each of wheat, chickpea, pea, mustard and barley and elemental bioaccumulation, metabolite concentration in component parts of chickpea. Its seeds were sown separately in earthen pots. Ca, K, P, total N, protein and ether extract contents were decreased in all the four component parts of the chickpea i.e. root, stem, leaf and seeds while Na, Fe and carbohydrate content increased. Concentration of sulphate was higher in stem followed by seed, leaf and root. Ash content was higher in the stem of effluent treated cultivar. There was no uniformity in the response exhibited by both the cultivars of all the five rabi crops in the accumulation of Mg, Pb, Cr and Zn in the straw. Earlier in 1990 from the same place, Bahadur and Sharma reported the effect of three effluents of different industries, Indian Turpentine and Rosin Co. Ltd., Western India Match Co. Ltd. and Camphor and Allied Products Ltd. on wheat. Percent germination decreased significantly in the treatment upto sixth day but the decrease was maximum on the first day. The effluent had inhibitory effect on seedling growth. The root and shoot length decreased significantly upto eighth day. Another experiment was conducted by Singh and Singh (1997) to study the effect of turpentine factory effluent and Zn on germination, seedling height, fresh and dry weight, chlorophyll content of Cajanous cajan (pigeon pea). Results revealed that higher concentration caused deleterious effect whereas lower had beneficial. Augusthy and Sherin (2001) at Annapuram, performed a petriplate experiment to study the effect of rubber factory effluent on green gram and reported that above 50% effluent retarted germination percentage while upto 50% favoured seedling growth. Length of root and shoot system and number of lateral roots was also increased by low concentration. Aziz et ai (1996) reported from Aligarh while conducting a field trial at Mathura Oil Refinery on wheat, triticale, chickpea. Lens culinaris (lentil), pigeon pea Rc^•iew of Literatuic

and summer moong. Crop, when irrigated with wastewater showed an increase in seed yield except in summer moong. Aziz et al. (1993a) in a pot experiment on green gram studied the effect of treated Mathura Oil Refinery wastewater on nitrate reductase activity (NRA). A linear increase was noted from 15 to 25 DAS and then the activity decreased. Aziz et al. (1993b) again tested the performance of lentil with N15P30K40 and no fertilizer. Treated effluent proved better as compared to ground water. Treated effluent with fertilizer further enhanced growth parameters. Inam et al. (1993) also compared the effect of refinery effluent and ground water on triticale and wheat. No significant difference was noted in percentage of seedling emergence under both . Siddiqui etal. (1994) while using the same water studied the response of green gram. It enhanced leaf number, dry weight, seed number and pod number. Effluent with fertilizer proved more effective as compared to ground water for vegetative growth only while it decreased the seed yield. Considering the same source of water, Samiullah et al. (1994) observed enhanced leaf number and dry weight in a field experiment on wheat. Yield characteristics and final yield ha'' of the crop were also enhanced. Aziz etal. (1994) again tested triticale and wheat and noted beneficial effect. They also performed a field trial in 1995 on four cultivars of wheat and observed beneficial effect on growth and yield but the effluent resulted in lower protein and carbohydrate content of the grain. Aziz et al. (1996) while taking the berseem {Trifolium alexandrium) under N25P100K50 and no fertilizer observed enhanced leaf number and dry weight. Treated effluent applied with fertilizer enhanced growth parameters. Aziz et al. (1998) during another study on frequency of irrigation and the productivity of triticale further observed the beneficial effect on growth, yield and quality of grain with increased number of irrigations. In 1999, maize and mustard {Brassica juncea L. Czern & Coss.) were irrigated and higher values for plant height, leaf area, fresh and dry weight and yield in maize and better shoot and root length, leaf number, shoot fresh and dry weight and yield in mustard were obtained. Effect of wastewater on protein and carbohydrate contents of maize seeds was non-significant whereas oil content and oil yield in mustard increased significantly. From the same laboratory, Hayat et al. (2000) also performed a field trial on the effect of refinery effluent on growth, yield and level of heavy metals in the

10 Review of Literature

seeds of mustard as well as in the soil. It enhanced plant growth, fresh and dry weight while yield characteristics did not show significant changes. Irrigation with both waters increased the level of Cr, Zn, Fe and Mn in the seeds but it was more with wastewater. Also at Aligarh but in different laboratory Ajmal and Khan (1983) studied sugar mill effluent on Phaseolus aureus (kidney bean) and Peimisetum typhoides (millet). Germination was quickest in the control and with 25% effluent. Ajmal et al. (1984) also assessed the impact of Glaxo Laboratories (India) Ltd. effluent on kidney bean and Pennisetum glaucum (pearl millet). 100% effluent decreased the germination of kidney bean, while pearl millet showed an increase as compared with control. 25% effluent increased growth of kidney bean whereas pearl millet showed better response with 50, 75 and 100% effluent. Ajmal and Khan (1984) while taking wheat and pea with breweries effluent noted 100% germination under 25, 50 and 75% effluent, while it was 80% and 90% for pea and wheat, respectively in 100% effluent. They also tested the suitability of Hindustan Lever Ltd. and its soap splitting unit effluent on pea and mustard in 1984b. In 1985a, they collected textile factory effluent and applied on kidney bean and Abelmoschus esculenius (lady's fmger). 50% effluent proved best for both the crops followed by 25, 75 and 100%, Germination was delayed in 100% effluent and it was 90%o under 75% effluents. Ajmal and Khan (1985b) also observed the impact of electroplating factory effluent on Dolochos lablab (hyacinth bean) and mustard. Delayed germination and retardation of root and shoot length were observed with increasing concentration, while germination was totally inhibited at 1.5% in mustard seeds. Fresh and dry weights of hyacinth bean increased upto 0.2% effluent. The optimum growth of hyacinth bean was observed with 0.1% effluent. They were of the opinion that only very diluted effluents proved favourable for plant growth.

Singh and Bahadur (1995) conducted a petriplate experiment to study the effect of distillery effluent on rice, wheat, black gram, green gram, pigeon pea, lentil, mustard, soybean, maize and chickpea. In 100% concentration no germination was observed while it was normal in 20%, whereas green gram germinated normally in 50%. On the other hand wheat was more sensitive and showed no germination at 50%, while reduced germination in lentil was observed at 50%. Ghosh etal. (1999) at

II Review of Literature

Patna also studied the germination under distillery effluent. It increased at 75% effluent in chickpea and pea and at 50% in mungbean. Distillery effluent was also studied earlier by Goel and Mandavekar (1983) at Karad on clusterbean. Seeds were germinated in pots and watered with 10, 25 and 50% effluent just after the emergence of seedlings. After 20 days, nodule number was equal in all the sets, but after 40 days it increased at 10% effluent while 50% effluent gave the least nodule number. N content was maximum at 50% and minimum in control. They concluded that 10% effluent can be effectively used. Also on distillery effluent Somashekar et al. (1992) at Bangalore, studied the response of cow pea and fenugreek. Germination, survival percentage, reduction in root and shoot length, vigour index and fresh and dry weight of both the crops decreased with increasing effluent concentration. They also concluded that properly diluted effluents could be used. Similarly, Kannabiram and Pragasam (1993) at Pondicherry taking the same effluent studied black gram. At 2.5% effluent they observed higher germination percentage and seedling growth. Khan and Dhaka (1996) at Ghaziabad, recommended the use of Simbhaoli Sugar Mill and distillery effluent to grow Saccharum officinarum (sugar cane), wheat, maize, mustard and pea economically after its proper dilution. Kannan (2001) at Periyakulum Theni, in petriplates also studied the effect of distillery effluent on green gram and millet. 1% effluent gave the highest values for germination percentage, shoot length, root length and vigour index of both the crops. In control seeds exhibited 100% germination while no germination was observed in 100% effluent. Ramana et al. (2002) evaluated the manurial potential of three distillery effluents i.e. raw spent wash (RSW), biomethanated spent wash (BSW) and lagoon sludge (LS) vis-a-vis recommended fertilizers (NPK + farmyard manure) and a control in a field study. The effluents increased total chlorophyll content, crop growth rate, total dry matter, NPK uptake and seed yield, but inhibited nodulation and decreased nitrogen fixation in groundnut. Among the three distillery effluents, BSW produced highest seed yield followed by RSW and LS. However, distillery effluent did not influence protein and oil contents. Kumawat et al. (2001) at Ujjain, while taking wheat and chickpea and testing in 0, 25, 50, 75 and 100% base and caustic yellow dye effluent noted that higher

12 Review of Literature

effluent concentration decreased the germination, root and siioot length and dry matter production of both the crops while lower concentration (25%) showed marginal increase. Ramasubramanian et al. (1993) at Sivakasi studied the impact of match and dye industry effluent on growth and metabolism of mungbean. Seeds were soaked for 2 hrs in diluted effluents (10-40% v/v) and grown in sand cuhure. With increase in concentration germination percentage, seedling growth, plant fresh and dry weight, chlorophyll 'a' and 'b', leaf soluble protein and in vivo nitrate reductase activity decreased while leaf L-proline increased. It was found that dye industry effluent was more toxic than match industry. Pronmurugan and Jayseelan (1999) at the same place conducted a petriplate experiment to evaluate the effect of fireworks and dye industry effluent on germination and growth of Typha angnstate (cuttail). Reduction of root and shoot morphology, biomass accumulation of seedlings was observed under higher effluent concentration. Shukla and Pandey (1991) obser\'ed the impact of oxalic acid manufacturing plant wastewater on maize, mungbean and chickpea. At 25%, seed germination of maize, mungbean and chickpea was 86, 32 and 55%, while at 50% it was 52, 12 and 15% respectively but in control it was found to be 100%i. 10 day old seedlings of maize, mungbean and chickpea showed decrease in height at 25% and it was found to be 5.1, 0.7 and 2.6cm, respectively as compared to control. Saha et al. (1994) at Shantiniketan, studied the behaviour of radicle in petriplates. Seeds of rice, mustard, lentil, green gram, chickpea and pea were soaked in carbon black factory and chemical factory effluents. They observed various degrees of phytotoxic symptoms and concluded that effluent caused deleterious effect on growth of radicle. Chidaunbalan et al. (1996) at Tuticorin performed an experiment to evaluate the effect of wastewater on green gram and black gram. 10% effluent proved effective in promoting germination, growth, chlorophyll and protein content. Murty and Raju (1982) At Waltair conducted a study on the effect of alum factory effluents on rice, green gram and mustard. Effluents were collected from Costal Chemicals dried in the laboratory, powdered and made test solutions of 25, 50,

13 Review of Literature

75 and 100%. In rice, at 25 and 50% shoot inhibition was less than root while in green gram shoot inhibition was more than root at 25%. At 25%, root and shoot growth was severely inhibited in mustard. Further inhibition of shoot growth and total inhibition of radicle emergence was observed in rice at 75 and 100%. Total inhibition of shoot and further reduction in root growth was noticed in green gram. In case of mustard, further reduction in growth of shoot and root was recorded at 50%, but complete inhibition of shoot and further reduction in root growth was observed at 75 and 100%. Balashouri and Prameeladevi (1994) tested tannery effluent on pigeon pea and sorghum. At \0% legume showed an optimum increase in seed germination, seedling growth, chlorophyll content and biomass whereas sorghum showed better response at 5%» effluent. Similarly, Karunyal et al. (1994) conducted petriplate as well as pot experiments on rice, Acacia holosericea (condelsbrawattele) and Leucaena leucocephala (subabul). Germination was inhibited at 25 and 50%) effluent and prevented by 75 and 100%. Some other crops like Gossypium hirsutum (cotton), black gram, cowpea and tomato {Lycopersicon esculenium) were also observed with the same effluent for 10 days and it was noted that leaf area, biomass, chlorophyll and protein contents increased at 25%. effluent. Arora and Chauhan (1996) at Agra reported significant reduction in germination percentage, length and total biomass in barley. Bera and Bokaria (1999) reported that 10% effluent did not significantly reduce seed germination in mungbean but in 50%, germination was 64%i compared with 96% in control. Early seedling growth, fresh and dry weights were better at 2.5%». Irrespective of concentration, chlorophyll 'a', 'b' and total chlorophyll decreased in 6 day old seedlings. They concluded that tannery effluent can be utilized as a liquid fertilizer only for certain crops at 2.5% dilution level. Thukral (1989) at Amritsar, performed a pot experiment to study the effect of tailings water irrigation on the biomass of green gram, cluster bean, millet, wheat, barley and mustard and reported decrease in dry weight of different plant parts and total dry weight. The crop worst affected was mustard in which total plant dry weight and dry weight of the fruits decreased. However, dry weight of the fruits increased by 50%o in clusterbean. 25.6% increase in dry weight was also recorded in the spikes of wheat. It was concluded that regular irrigation with tailings water retarded the growth

14 Review of Literature

of crop plants. Trivedi and Kirpekar (1991) at Karad carried out an experiment on soybean and mungbean. Dairy waste increased the ash, Ca, N and P content of both the crops. In case of soybean, P content increased in 10%, 25% and 50% but decreased in 100% effluent. At Bhavnagar also dairy effluent was tested by Prasanna et al. (1997) in a petriplate experiment conducted on green gram and black gram. Only under 15% effluent germination percentage, seedling growth and pigment content were increased while increasing concentration proved deleterious. Earlier Pathak et al. (1992) at the same place utilized pre-treated effluent of manufacturing plant of Excel India Ltd. for raising agroforestry. Subabul was found to grow well on soil being irrigated by the effluent. Srikantha (1998) at Bangalore conducted a pot experiment on dairy effluent on french bean and Amaranthus (amaranthus). Germination decreased with increase in concentration. Dry matter yield of both the crops was recorded highest in control and lowest in undiluted effluent. Uptake of plant nutrients decreased with increase in effluent concentration as compared to control. Heaton et al. (2002) evaluated the influence of two rates of dairy pond effluent application on spatial root distribution of Salix viminalis PN386 (commonosier) and NZ1295 and Eucalyptus nitens (shining gum). It was found that spatial distribution was influenced by effluent rate, with greater quantities of both fine and coarse roots in the top soil horizons with the higher effluent rate of 300m^ ha'' compared to 150m'' ha''. Gupta and Nathawat (1992) at Jaipur, took various concentrations of textile effluent and the effect on germination and seedling growth of pea was observed. Effluent exerted toxic effect on seed germination and seedling growth. Root and shoot length and biomass decreased with increasing concentration. But root was adversely affected than shoot. Earlier using the similar effluent in 1988 Annon at Naroda, performed a field experiment for the successful cultivation of jawar, Penmsetum americamim (bajra), okra, tomato and chilly {Capsicum annum). On the same lines Singh et al. (2001) at Jodhpur suggested that some forest tree species can be established successfully using textile industrial wastewater in arid region. Vijayakumari et al. (1993) at Coimbatore evaluated the effect of soap factory effluent on pearl millet Stapf Hubbard, finger millet {Eleusive coralana L. Goertn),

15 Review of Litciiituie green gram and black gram. Seeds were soaked in petriplates and after 24hrs, the imbibed seeds were allowed to germinate on germination towel. At 100%, germination percentage and growth of shoot, root and lateral roots were decreased in both pearl millet and finger millet whereas in pulses seed germination and seedling growth were totally suppressed. Upto 5%, cereals showed an increase in seedling growth but the pulses have registered maximum growth when treated with 2.5% dilution. Srivastava ei al. (1995) at Jabalpur, investigated the effect of ordinance factory effluent on pea. It was highly deleterious for the germination and early growth. The deleterious effect was increased with increasing concentration. Jabeen and Abraham (1997) at Thiruvanthapuram, in petriplates, noted the effect of Hindustan News Print Factory effluents on seed germination and seedling growth in Cassia iora (foetid). Cassia roccidntalis (coffeeweed) Vicia faha (bakia) and cowpea. The effluent caused stimulatory effect in most of the parameters studied. Adverse effect on germination and seedling growth were negligible. Klimakhin et al. (1998) in a separate study conducted in petriplates and pot to evaluate the effect of sugar factory effluent on wheat, oat, lucerne (Medicago sativa), pea and sugar beat {Beta vulgaris). It was found that germination energy and germination capacity were in all cases equal or slightly superior to that of control. But in pot experiment, the effluent had beneficial effect on plant growth and development. Tiwari and Tripathy (2001) also noted the effect of Balrampur Chinni Mill's effluent on pea var. Asanji and Arkel. In the control, 100% seed germination was achieved on 6"^ day while seeds treated with 25% effluent showed 95% and 85% germination in Arkel and Asanji respectively on 1^ day. Maximum inhibition was observed at 100% effluent in both varieties. Kumar (2000) at Madhubani, conducted an experiment to study the influence of periodic watering with Chakia Sugar Mill's effluent on polygenically controlled characters of barley. Results revealed that each parameter was reduced when irrigated with effluent. Abasheeva and Revenskii (1992) at Uan-Ude (Russia) conducted a pot experiment to study the impact of seleginsky cellulose and cardboard mill effluents on productivity and chemical composition of Oat, rape {Brassica napus) and pea. Crops

16 Review of Literatuic

were grown in pots containing alluvial meadow or grey forest soil and irrigated with clean water or purified wastewater. The wastewater increased dry matter yield of oats on both soils whereas dry matter yield of pea on grey forest soil and did not affect those of rape on either soil or pea on alluvial meadow soil. There were no adverse effects on chemical composition or food value of crops. Karpate and Choudhary (1997) at Nagpur, performed an experiment to study the impact of thermal power plant waste on wheat. The crop was grown in fly ash amended soil (50, 70 and 90%) and irrigated with various concentrations of fly ash water i.e. 25, 50, 75 and 100%. At lower concentrations the fly ash water and fly ash had stimulatory effect on crop, while higher concentrations showed deleterious effect. Das et al. (2000) at Dhanbad, conducted a petriplate experiment to evaluate the effect of Chadrapur Power Plant's fly ash pond effluent on peas and Vicia sativa (common vetch). At 25% effluent the germination rate was stimulated in both legumes. But at 50%, only 0.5% enhancement was noticed in pea. Shoot growth was promoted upto 50% in both the crops. Maximum growth index was recorded with 50% effluent. It was concluded that diluted effluent may be used for better plant growth. Shetty and Somashekar (2000) studied the germination and growth of kidney bean when irrigated with effluent of Bharat Heavy Electricals and reported that diluted effluent showed better percentage germination, growth and chlorophyll content. At 75-100% concentration drastic reduction was noticed. The effluent contained several plant nutrients and therefore, as pointed out by them, could be recommended for irrigation at lower concentrations. Crowe et al. (2002) at Alberta (Canada) studied the effect of major oil and industrial companies released consolidated tails (CT) water on plant colonization and germination and post-germination growth of some terrestrial and aquatic plants. To check viable options for reclamation hummack-wetland system had been constructed, in addition, natural wetlands had been established as a result of seeping of the effluents. Vegetation surveys revealed that constructed wetlands and consolidated tails had low biodiversity and were not invaded by many aquatic plants. Effluent had an inhibitory effect on germination of tomato, clover {Trifolium pragiferum), wheat, rye (Seca/e cereal), pea, reed canary grass {Phalaris arunadinacea) and loblolly pine

17 Review of Literature

{Pimts taeda). Effluents from constructed wetland and CT were responsible for delay in germination and reduction in fresh weight of seedling of tomato, wheat, clover and loblolly pine. Stehlik (1986) at Harlickuv Brad (formerly Czechoslovakia) studied the seedling growth of Sinapsis alba (white mustard) irrigated with brewing starch factory and canning factory effluents and reported that starch wastewater was found to be usable after dilution, while distillery wastewater showed adverse results even if diluted. Again in 1987 he performed a petriplate as well as pot experiments to investigate the effect of yeast plant wastewater on white mustard and found that undiluted wastewater had little or no inhibitory effect on germination, whereas separate fractions of wastewater reduced germination. Dry and fresh yields of pot grown plants were increased by wastewater. Baumgartel and Fricke (2000) at Hanover (Germany) performed a field trial to monitor soil N levels and uptake of N and K by winter rape (Brassica napus biennus) cover and catch crops after the application of wastewater from starch potato processing factories in late summer. They concluded that wastewater may be used as liquid fertilizer. Sundari and Kanakarani (2001) at Kodaikanal assessed the impact of pulp unit wastewater. The analysis showed that the partially treated effluent had adversely affected the soil fertility and crop production. Similarly earlier at Florida (USA) Walsh et al. (1991) exposed vascular wetland plants to effluents discharged from a pulp mill in addition to coke plant and wastewater treatment plant. There was no effect of effluent on germination but growth rates were reduced significantly in most cases. Murillo et al. (2000) at Cabrera (Spain) conducted a field experiment to examine the effect of drip irrigation using wastewater from olive industry. Wastewater caused decrease in leaf water potential, stomatal conductance to H2O and the photosynthetic rate in olive trees, after 15 days of irrigation. Wastewater significantly reduced olive yield as compared to control. Sedykh and Tarakanov (2000) performed a petriplate experiment to study the effect of oil and gas drilling waste on some woody plants. Results showed that small doses of drilling waste

18 Review of Liteiatuie

(below 10%) can stimulate germination and sprouting intensity but doses greater than 20-25% reduced it. Barman et al. (2001) at Lucknow, reported the impact of electronic component manufacturing unit effluent on soil and plants. Effluent showed higher accumulation of metals in plant parts of water hyacinth and Marsilea sp. (four leafed clover). In 2000, Salgare and Acharekar studied the effect of industrial pollution on weeds of Kalu river. Industrial pollution of Kalu river inhibited all the parameters studied under growth performance of five weeds. In 1991, also Salgare and Andhyarujina at Mumbai, evaluated the effect of polluted water of Fatal ganga on the mineral contents of its bank vegetation. Out of the four species collected Croiolaria retusa (rattle weed) was found to be most sensitive species as it showed maximum inhibition in the mineral contents while Argemone maxicana (prickly poppy) and Leucas aspera (dronapushpi) were found to be more resistant. Prashanthi and Rao (1998) at Hyderabad, performed a petriplate experiment to evaluate the effect of industrial effluents and polluted water on seed germination of some crops. The effluents and wastewater were found to be unsuitable for irrigation purpose. Again, they (Prashanthi et al., 1999) performed an experiment and reported that germination and dry matter yield of some crops treated with polluted well water and soil were reduced as compared to control. Dhafer et al. (2000) conducted an experiment to investigate the impact of complementary wastewater irrigation using an infiltration-percolation process on the growth of durum wheat (Triticum durum) and observed an increase in yield. 2.2 Effect of fly ash on crop plants Coal based thermal power plant stations have been the major source of power generation in India and nearly 15-30% of the total amount of residue generated during coal burning constitutes the fly ash. Its particles when dumped are responsible for serious problems to human and animal health (Page et al., 1979; Borm, 1997) as well as to the higher plants. Impact of fly ash is however, very complex therefore, its assessment has been addressed for both the negative as well as the positive effects on plants. Partly it may be a potential nutrient source if the soil is amended with a limited quantity making it eco-friendly during ecological recycling, whereas its utilisation in

19 Review of Literature

some industrial purposes has also been established (EPD 1993). Unfortunately, the major part of fly ash is disposed off in unmanaged landfills or lagoons, which serve as major source of environmental pollution. To overcome this menace, revegetation trials of fly ash landfills have been conducted involving appropriate blending of fly ash with organic wastes and the application of nitrogen fixing organisms (Cheung et ai, 2000; Rai et al., 2000; Vajpayee et ai, 2000). It may be pointed out that fly ash is enriched with macro and micro nutrients, which may enhance the plant growth specially under nutrient deficient soils (Plank and Martens, 1974; Martens and Beahm, 1978). However, the main constraint in the use of fly ash is the high alkalinity and its salt contents, which may depress the plant growth and may cause the deterioration of soil (Hodgson and Holliday, 1966). Despite this, variety of vegetables, millets, cereals and trees have been observed to grow successfully in soils amended with fly ash (Lisk et al., 1979; Singh and Singh, 1986; Sikka and Kansal, 1995; Kumar et al., 1996; Khan and Khan, 1996). The review dealing with fly ash effect has been divided into leguminous and non-leguminous crops in the present thesis. Matte and Kene (1995) conducted a field trial at Nagpur and observed the yield performance of various kharif and rabi crops. Cotton, sorghum, groundnut, soybean, green gram and rice were grown in kharif (monsoon) and wheat, gram and mustard in rabi (winter) season. Crops were given 0-15t fly ash ha"' along with 0, 75 or 100% of recommended rates of NPK fertilizers. Application of lOt fly ash ha'' gave the best results, increasing seed grain yield by 7-30%. In 1997, Kuchanwar et al. also studied the effect of graded doses of fly ash and fertilizers on nutrient content and uptake of groundnut grown with 0, 5, 10 or 15t ha'' along with N25P50 or N18.75P37.50. Application of fly ash and N, P fertilizer separately and in combination significantly increased the N, P and Mg contents in plants. The highest N, P, K, Ca and Mg contents were recorded with N25P50 Vi'hile highest uptake of all nutrients was observed when lOt fly ash ha"' was applied. During the same year Karpate and Choudhary while working on wheat also reported beneficial role of fly ash when given in lower concentration. Bhaisare et al. (2000) at Nagpur conducted an experiment on green gram with

20 Review of Literature

the NoPo, N18.75P37.50 and N25P50 and reported that highest yield of grain and straw along with highest content and uptake of nutrients, crude protein and test weight were recorded with lOt ha'' fly ash. Again crop responded well to higher dose of N and P fertilizers for yield, quality, nutrient content and their uptake. Kalra et al. (1997) at New Delhi conducted a pot experiment to evaluate the effect of fly ash amended soil on germination and stand establishment of wheat, chickpea, mustard and lentil during the winter season and rice and maize during the summer season. The applied fly ash concentrations were 0, 10, 20, 30 and 40% for rabi crops and 0, 5, 10, 15 and 20% for summer season crops. Fly ash amended soil delayed the germination. They also observed that summer season crops were comparatively less sensitive to ash for germination than winter season crops. Among the winter crops, mustard was most affected in terms of germination as well as stand establishment. Again in 1998, they studied the effect of fly ash on growth and yield of wheat, mustard, rice and maize. Fly ash was added @ 5, 10, 15, 20 and 50t ha''. The seed yield in maize and mustard increased with lOt ha'' fly ash. On the other hand the yield of wheat increased upto 20t ha'' and it declined thereafter. In case of paddy yield addition of lOt ha"' of ash could not increase the seed yield as it was similar to no fly ash. Barman et al. (1999) at Lucknow, conducted a field experiment with fly ash amended soil. They observed accumulation of heavy metals in turnip, cabbage, Daucus carota (carrot), raddish, Spinacea oleracea (spinach), peas, coriander, Lactuca sativum (lettuce), tomato, Solatium melongena (brinjal), chickpea and mustard receiving fly ash of a thermal power plant. In the edible parts Cu, Zn and Pb concentration was within permissible limits, whereas Cd, Cr and Ni was more. At the same place, Srivastava et al. (1995) observed the 10% fly ash amended soil showed a marked increase in plant growth of lettuce while 20 and 30% retarded it. Some experiments related to fly ash-soil amendment were also carried out at Aligarh to evaluate the impact on various crops. Mention may be made of Khan and Khan (1996) increase in growth, yield, carotenoids and chlorophylls of tomato with 40-80% fly ash, being optimal at 50 or 60%; Khan et al. (2000) beneficial effect for growth, seed germination and metal uptake in pigeonpea, black gram and okra upto

21 Review of Literature

75.g kg'' soil; Siddiqui et al. (2000) who reported increase in various parameters of chickpea under 40% fly ash; Khan and Abdussalam (2001) beneficial effect of lower and medium concentrations on some ornamental plants; Khan et al. (2001) beneficial effect on some physiological parameters at lower doses upto 20-30g fly ash ha' soil in barley and wheat; Ahmad and Khan (2002) increase in fresh and dry weight of pea at 20% fly ash level, whereas fresh and dry weight of nodules under 20-80% ash level; Ahmad and Saeed (2002) increase in leaf number, fresh weight plant'' and total biomass in linseed at 40% fly ash level; Raghav et al. (2003) evaluated the effect of fly ash and ceramic dust as a soil amendment and reported that 20% fly ash proved better for grov^h and yield performance in tomato; Singh and Khan (2003) reported the same concentration of fly ash for better growth of mustard cultivars under conditions of induced drought; and Singh and Siddiqui (2003) significant increase in plant growth and yield of rice under 40% fly ash level.. Elseewi etal. (1978) at California (USA) carried out a green house experiment to study the availability of sulfur from fly ash. Two series of experiments were conducted. In first one, fly ash was mixed with soil at the rate of 0, 0.25, 0.50, 1, 2, 4 and 8% by weight. Sufficient amount of sulfur free-mixed fertilizer was added to each pot at N, P and K rates equivalent to 400, 250 and 300mg kg'' for alfalfa and 250, 160 and 200mg kg'' for bermudagrass respectively. In the second experiment, fly ash and gypsum were mixed with soil @ 25, 50 and lOOmg S kg'' soil. N and P were added at the rate of 200 and lOOmg kg'' and seeds of turnip and white clover were sown in the pots. Fly ash improved S deficiency in the soil and maximised the yield of alfalfa and bermudagrass. The increase in yield was accompanied by an increase in S content of the plant tops from deficiency level to sufficiency level. Significantly the yield and S content of turnip and white clover were equally improved indicating the availability of fly ash derived-S, equivalent to that of gypsum-S. Aitken and Bell (1985) at St. Lucia Qld. (Australia) studied the uptake and phytotoxicity of B present in fly ash. French bean and Rhodes grass {Chloris gayer) in one experiment grovm in fly ash-acid washed sand amendment (5 and 10% by weight) and in another experiment, grown in 0, 15, 30, 70 and 100% fly ash-sandy loam soil amendment. Water holding capacity of the soil was increased by fly ash.

22 Review of Literature

however, adding large quantity of untreated fly ash resulted in poor plant growth due to boron toxicity. Rhodes grass tolerated higher boron toxicity by absorbing less elements than French bean. In their opinion phytotoxicity of boron could be a major problem in establishing vegetation on ash. Shukla and Mishra (1986) at Kanpur conducted a petriplate experiment on fly ash extract and studied growth and development of com and soybean seedlings. In 0.5 to 1.0% fly ash extract, no significant deviation in germination, root and shoot growth, fresh and dry weight and pigment content in both crops was noted. However, higher concentration adversely affected these parameters. The treated seedlings accumulated more amount of elements in the root and shoot. In 1986, Mishra and Shukla they fiirther determined the elemental composition of corn and soybean. 5% fly ash level increased plant height and biomass, while higher concentration proved deleterious. Fly ash showed no change in P, K or Ca content of roots, shoots or seeds, but increased their B, Cu, Mn and Zn content. 25% fly ash level increased B content in roots of both crops. Srivastava et al. (1995) rehabilitated the fly ash dump yards at Panki Thermal Power Plant, Kanpur by growing six nitrogen fixing and four non-nitrogen fixing tree species. Suitable treatment of fertile soil (50%), FYM (2.0kg) and DAP (150g) was imposed in each pit taking fly ash in a pit as control. Survival percentage, height and diameter were noted every year. Observations during third year under treated conditions revealed that the top performing tree species were subabul Acacia nilotica (acacia), Albizzia lebbek (lebbek) and Pithecllobium duke (kamat siri). The overall best performer under treatment was subabul and without any treatment was acacia. They suggested to rehabilitate fly ash dump yards with nitrogen fixing tree species. Benes and Mastalka (1987) at Prague (formerly Czechoslovakia) studied the element uptake by plants from soil enriched with power station waste. Bakla, french bean, mustard, pea, barley, cabbage, maize, flax, oats, fodder peas and buck wheat were studied in the laboratory and french bean and barley under the field conditions. They reported that soil with high content of power plant ash was proved unsuitable for crop cultivation. Menon et al. (1993) at Savannah (USA), investigated the suitability of fly ash

2.3 Review of Literature amended compost prepared from grass and amended with fly ash collected from coal fired power plant, as a manure for mustard, coUard greens, string bean, bell pepper and eggplant grown on three compositions namely, soil alone, soil amended with composted grass clippings, and soil amended with mixed compost of grass clippings together with 20% fly ash. The fly ash amended compost enhanced dry matter yield of coUard greens and mustard only but string beans, bell pepper and egg plant showed no significant increase. Dzeletovic and Filipovic (1995) at Zemem (Yugoslavia) studied autumn rye, lucerne, barley and winter rape on power plant ash and bottom slag deposits. According to them with the application of conventional agricultural techniques, good grain seed quality was achieved on various parts of the ash bottom slag deposits. However, it could be achieved with high fertilizer levels given at the rate of

N228P90K90. Lai and Mishra (1996) at Ranchi evaluated the effect of fly ash on nodulation in a pot study. Seeds of soybean were inoculated With Bradyrhizobium japonicum and grown under 0, 4, 8, 16, 32 or 100% fly ash and 0, 50, or 100% of recommended NPK fertilizer doses. Nodule number was highest with 8% fly ash level, but further increase in fly ash concentration decreased it. However, NPK application increased the nodulation. Nodule dry weight plant'' was highest with 8% fly ash + recommended NPK rate. Lai et al. (1996) also at Ranchi, studied soybean plant, grown in pots containing acid alfisol (pH 4.9) amended with different rates of fly ash. The highest dry matter yield of soybean was obtained at 16% fly ash level and a significant reduction in plant growth was observed at levels higher than 16%. Khandkar et al. (1996) at Pantnagar while growing rice cv. Jaya soybean cv. PK-327 and black gram cv. Pant U-30 considered a combination of clay loam soil with unweathered coal fly ash as 0, 2, 4, 6, 8, 12, and 20%. Crop yield was increased upto 6% except for soybean at 20%. Straw yield of rice and soybean was increased by 4% fly ash and straw yield of black gram by 6%. According to Patil et al. (1996) application of fly ash @ 20t ha'' resulted in higher yield of sunflower when compared to control at Raichur. Baskaran et al. (1998) in TamiJnadu conducted a field experiment to observe

24 Review of Literature the effect of fly ash on rice, greengram, groundnut and brinjal. One crop of groundnut was grown in kharif 1996 under 2.5 and 5t ha'' with or without FYM at the rate of 12.5t ha"'. In addition, two crops of groundnut were grown in summer 97 and kharif 97 at 5 and lOt ha'' of fly ash. Rice showed marginal increase in the number of tillers at lOt ha"'. Green gram and groundnut were not affected adversely in germination, crop stand and nodule number while pod yield was increased under 2.5t ha'' fly ash + 12.5t ha"' FYM. Slight increase in growth of brinjal was observed at 2.4t ha''. Jha et al. (1998) at Dhanbad, carried out a field experiment for biological reclamation oiXow lying areas and waste lands, by applying fly ash in bulk through afforestation, teak nursery raising and cultivation of crops. Indian rosewood was grown under the scheme of afforestation in Murshidabad. In another experiment teak seeds were sown in plots after proper levelling, ploughing and mixing of fly ash. Growth rate of the plants in the ash filled reclaimed soil was comparatively faster than the plants grown on normal soil. In case of teak, germination rate of seeds, height of seedlings and their collar diameter was higher with increasing dose of fly ash. Tripathi et al. (1998) evaluated its impact in light and shade environment on growth and chemical response of Albizia procera (acacia) and acacia gum. Seeds were germinated in pots under 10, 20 and 30% fly ash-soil amendment. Lower concentration favoured the growth of both the tree species, however, higher concentration had adverse effects. Sarangi and Mishra (1998) at Buria (Orissa) evaluated the soil amended with 15% fly ash in agriculture as nutrient supplement in groundnut, lady's finger and radish. Root length, shoot length, biomass, leaf number, total leaf area plant'" and yield were mostly enhanced in fly ash amended soil.

Vallini ei al. (1999) at Verena (Italy), in a pot experiment studied the growth response of bakla with different amendments of coal alkaline fly ash, co-composted fly ash and lignocellulose residues. At the rate of 5 and 10%, in both soils, neither fly ash alone nor co-composted fly ash extracted any negative effect. Plant biomass production was not influenced in either clayey or sandy soil. In a field experiment effect of fly ash-soil amendments on yield and quality of soybean and wheat was studied by Kumar et al. (1999). Fly ash-soil amendment (4-16%) increased the yield of both crops. Application of 50 and 100% NPK showed similar results especially at

25 Review of" Literature higher levels. At the same place they (Kumar et a!., 1998) had reported beneficial role on rice, when given in lower concentration. The application of 8% fly ash + recommended NPK rate proved highest grain yielder of 4.85t ha' which was not significantly different from 4.63t ha'' obtained with 4% fly ash + lOt FYM + 50% of the recommended NPK rate. Das and Jha (2000) evaluated its effect on pigeon pea in petriplates. Seeds were germinated and studied under different concentrations of fly ash in water. There was slight stimulation in plant height and root length at 15-30% while at 45% germination and seedling growth were decreased. KeGong et al. (2000) at Henan (China), studied the effect of magnetized fly ash compound fertilizer. In a field trial, soybean was given 49kg magnetized fly ash compound fertilizer 667m' . Results indicated the increase in yield which was found to be higher in magnetized fly ash than same amount of NPK fertilizer and with non-magnetised fly ash compound fertilizer. The magnetized fly ash compound fertilizer improved soybean root nodule formation. Sriramachandrasekharan (2001) at Cuddalore, conducted a filed experiment to study the effect of lignite fly ash (LFA), gypsum, biodigested pressmud (BP), FYM and lignite humic acid (LHA) applied singly or in combination on groundnut. All treatment significantly enhanced pod number, 100 kernel weight, seed and haulm yield, protein content and nutrient uptake. Application of 7.5t BP ha"' + 1.2t LFA ha' + 200kg gypsum ha"' recorded highest pod number, 100 kernel weight, pod yield, haulm yield, oil and protein content and NPK uptake. Poonkodi (2003) at Annamalai studied the effect of lignite fly ash (LFA) on the performance of groundnut. Four concentrations of LFA were taken and recommended dose of NPK i.e. 17, 34 and 54kg ha'' was applied basally. Results revealed that 6t LFA ha'' registered maximum pod yield ahhough it was at par with 5t and 4t LFA ha"'. While considering the non leguminous crops, Korcak (1985) at Bethville (USA) studied the effect of coal combustion wastes used as lime substitute on nutrient of apple seedlings in three soils. Growth was reduced by 60% on the manor soil amendment with fly ash, applied at twice the lime requirement. Leaf N, P, K, Cu and Al were not significantly affected by treatments while Ca and N decreased.

26 Review of Literature

Singh and Singh (1986) at Varanasi conducted an experiment on rice. Three fertility levels of NPK; low (60, 40, 30kg ha''), medium (80, 40, 40kg ha'*) and high (120, 180, 60kg ha'') and four of fly ash (0. 10, 20 and 30%) were tested. Productive tillers, panicle length, test weight, number of grains panicle'' and filled grains panicle'' were significantly more in high fertility levels. Seed yield was also maximum in high fertility level which was significantly superior to medium and low fertility levels. 20% fly ash significantly increased the number of productive tillers hiir', length of panicle, number of grains panicle"' and test weight and decreased the unfilled grains. Joseph (1987) in USA studied the growth response of Agrostis tennis var. Highlander (colonial bentgrass), Festuca arundinacea var. Kentucky 31 (fescue), Lespedeza ameata (sericealespedeza) and soybean var. Brogg on fly ash amended strip mine soils. Seeds were sown in soil, mixed with 70t ha"' fly ash. Mean biomass production was markedly higher for each species in fly ash treated plots. There was production of numerous and apparently healthy root nodules in case of colonial bentgrass, indicating vigorous nitrogen fixation. Beresniewich and Nowosielski (1987) at Skierniewice (Poland) compared the fertilizing effect of brown coal ash with that of limestone. Upto 20t ha"' of ash or limestone were applied. Ash increased the vegetables yield. Wong and Wong (1989) at Kowloon (Hong Kong) studied the germination and seedling growth of Brassica parachinensis (Chinese flowering cabbage), and Brassica chinesis (Chinese cabbage). In sandy soil amended with 3 and 6% fly ash, germination was enhanced, while in sandy soil amended, with 12 or 30% and sandy loam soil with 30% fly ash significant reduction was noted. Dry weight production of crops was enhanced and the length of first leaves, shoot and cotyledons were found to be greater with 30% but reduced with 12 and 30% in both the soil types. In another experiment, they (Wong and Wong, 1990) also evaluated its effect on yield and elemental composition of the two crops which were grown in pots, filled with sandy and sandy loam soil amended with 0, 3, 6, 12 or 30% fly ash. Crop yield was highest at 30% fly ash level, except for Chinese flowering cabbage grown in sandy loam in which yield was highest at 12%. Yield of both the crops was significantly higher in

27 RevicAV of" Literature

sandy loam than in sandy soil. Lau and Wong (2001) also tested the use of weathered coal fly ash, to allevate the toxicity of manure compost. Addition of lagoon ash at the rate of 5% for immature manure compost and 10% for mature manure compost resulted in higher seed germination rate and root length growth of lettuce. Su and Wong (2002) at the same place, determined the amount of coal fly ash required to stabilize sewage sludge, without causing an adverse effect, on the growth of corn seedlings in a loamy soil receiving the ash-sludge mixtures amendment. Dry weight receiving 1:5 ash sludge ; soil mixture (v/v) was significantly higher than ].] soil mixing (v/v). The highest yield was obtained at 5 and 10% ash sludge mixture amended soil at 1:5 soil mixing ratio. Nevertheless, the yield at 35% ash-sludge amended loamy soil at 1:1 v/v was still higher than that of soil with fertilizer treatments. Singh ei al. (1994) studied the growth response and element accumulation in fodder pea. 2, 4 and 8% were mixed with soil m Im plots and seeds were sown. Low amounts of fly ash proved favourable for plant growth and it improved the yield. However, heavy metal accumulation was higher in plants grown in fly ash amended soil. Kenneth et al. (1995) at Aiken (USA) performed a green house experiment and evaluated the effect of fly ash/sewage sludge mixtures and application rates on biomass production of jawar. All mixtures of sewage sludge with fly ash generally increased plant growth and yield at 50:100t arcre"' but showed reduction in growth and yield at higher application rates. Mcload and Thomas (1997) also at Aiken studied the differential sensitivity of Nyssa aquatica (water tupelo) and Tcaadhtm distichuw (common bald cypress) grown under 0, 2.5, 5 and 10% fly ash-sand amendment. Growth of common bald cypress was not adversely affected by fly ash addition, however, biomass, basal diameter and height of water tupelo were reduced at the two higher fly ash rates. It was found that regeneration of water tupelo would be less successful than common bald cypress in wetlands containing fly ash. In a pot experiment suitability of fly ash for reducing heavy metal toxicity in maize plants was studied by Shende et al. (1995). They confirmed the suitability of 2% and 5% fly ash to calcarious soil as it showed better plant growth, while beyond

28 Review of Literature

5%, the crop growth was significantly reduced. On the contrary, acidic soil showed positive response to fly ash addition upto 20% but the resulted crop growth was comparatively lesser. Application of fly ash at 2-4% enhanced dry matter yield of paddy, while at 8% decrease in dry matter yield was reported by Sikka and Kansal (1995). Plant growth and food quality of barley, bronues and lucerne under 0, 25, 50, 100, 200 and 400t ha'^ of fly ash was observed by Hammermeister et al. (1996). They reported significant increase in yield of barley at intermediate rates of fly ash. Barley was grown by Sale et al. (1996) on weathered soil mixed with unweathered fly ash and observed an increase in plant height and grain yield under 6.25% and 12.5% fly ash, while at 25% there was no appreciable increase. At Hissar, Gupta et al. (1996) conducted a green house experiment to study the interaction of fly ash and phosphorus on yield and P uptake by wheat and observed decrease in grain and straw yield when fly ash was applied beyond 25%). At the same place Grewal et al. during the year 2001 reported that fly ash enhanced the grain as well as straw yield in pearl millet and wheat when grown upto 20%) fly ash. Uptake of N, P and K in grain and straw of both the crops was also higher under fly ash treated plots as compared to control. Kumars/a/. (1998) at New Delhi, on brinjal, potato, pea, tomato and cabbage; Thripathy and Sahu (1997-1998) at Berhampur on wheat and mustard; James et al. (1998) at Kharagpur on rice and sweet potato; Saxena and Asokan (1998) on cabbage, tomato, potato, pea and brinjal; Mandal and Saxena (1998) on paddy and soybean at Bhopal; Malewar et al (1998) at Parbhani on some forest and dry land fruit crops, also reported beneficial effect of fly ash when given in lower concentrations. Sugawe et al. (1997) at Parbhani (Maharashtra) studied the response of sunflower to the graded levels of fertilizers and fly ash. Five fertility levels namely Fi-FYM at the rate of lOt ha"'; F2-100% RDF (60:40:30); F3-75% RDF; F4-50% RDF and F5-F1 + F2 and five fly ash levels 0, 5, 10, 15 and 20t ha"' were considered. Application of FYM at the rate of lOt ha"' + 100%) RDF recorded better seed yield although it was at par with F2. Application of fly ash at the rate of lOt ha"' gave the maximum seed yield. Birajdar et al. (2000) also at Parbhani conducted an experiment to study the effect of fly ash and FYM on nutrient availability of soil and yield of

29 Review of Literature

sweet potato. There were three levels of fly ash (0, 5, 10 and 15t ha"') and two levels of FYM (10 and 15t ha''). The recommended dose, N6oP6oKi2o was given after planting. Application of fly ash increased the tuber yield significantly with each increment dose upto 15t ha''. FYM also increased the tuber yield. Masilamani and Dharmalingam (1999) in Tamil Nadu, studied the germination behaviour of teak drupes in fly ash added medium. The older drupes germinated and produced more and better quality seedlings than the fresh drupes. The best germination was observed in the fly ash + red earth + FYM mixture. Selvakumari et al. (2000) also in Tamil Nadu, studied the effect of integration of fly ash with fertilizers and organic manure on rice in Alfisols and reported an increase in yield at 20 and 40t ha'' of fly ash. The treatments receiving N, P and K as fertilizers or fertilizer + compost or fertilizer + Azospirillum recorded an increase in yield over the treatments without any manurial addition. The highest yield was recorded when fly ash applied at 40t ha"' in combination with fertilizer, compost znd Azospirillum. Gregorczyk (2000) at Szczecin (Poland), also observed its effect and the product of removing SO2 and NO from waste gases coming from power station, as a source of fertilizer, on yield and composition of spring rape. Fly ash was applied at 0.3 or 0.6kg 9kg'' capacity 'Mitscherlich' pot and N was applied at 0.5, 1.5 or 3.0g pot"'. No significant differences were found in the yield with different treatments. Application of 0.6kg ash pot"' reduced dry matter yield as compared to control. Sharma et al. (2001) at Dadri, reported the effect on soil health and yield of maize and rice under field conditions. The grain yield of maize and rice in fly ash treated plots was found to be increased. Tomato plant growth and yield under 1, 2, 3 and 4kg fly ash m"" applied to soil by broadcast or in rows was studied by Khan and Singh (2001). Plants grown in ash treated plots especially at 3 or 4kg dose, showed luxuriant growth and greener foliage. Growth and yield of three tomato cultivars were increased as compared to control. Naveen et al. (2001) at Annamalai, observed the effect of gypsum and lignite fly ash (LFA) as sources of sulphur on ragi {Elusive coracemo) cv. CO-12. The treatments were, control; gypsum at 80kg ha"'; 160kg ha"'; 240kg ha'', LFA at 2.It ha''; 4.2t ha'' and 6.3t ha"'. Gypsum at 240kg ha"' gave the highest values for total dry matter production, grain yield and straw yield.

30 Review of Literature

Rautaray et al. (2003) conducted a field experiment to study the direct effect of fly ash, organic wastes and chemical fertilizers on rice and mustard. The effect of fly ash on mean rice equivalent yield of the rice-mustard cropping sequence was highest when it was used in combination with organic wastes and chemical fertilizers. 2.3 Effect of NPK on chickpea {Cicer arietinum L.) Among various essential plant nutrients, N, P and K are considered to be of prime importance as these are absorbed and utilised in larger quantities. Therefore, a balanced dose of these nutrients in presence of specific biofertilizers can give much better results in leguminous crops. The requirement of these nutrients for different crops has been worked out and reported from time to time. Infact, there is sufficient literature available regarding this aspect for various leguminous crops (Subrahmanyam and Varshney, 1974; Paricha el al., 1983; Badole et al., 1991; Kushwaha and Singh, 1992; Singh et al., 1992; Singh et al., 1993; Yahiya and Samiullah, 1994; Patra et al., 1995; Rana et al., 1998; Kumar et al., 2000; Vyas et al., 2001; Bhat et al., 2002; Singh, 2002; Golakiya et al., 2002; Gundalia et al., 2002; Vijaybaskaran and Thirumurgan, 2002). In the following pages some of the important and relevant trials conducted specially on chickpea in relation to NPK were considered briefly. Because, the present study is not based only on fertilizer doses but it includes mainly the use of thermal power plant wastewater as a source of irrigation and nutrients in addition to the fly ash released from the same source. Javia et al. (1989) reported that seed yield increased from 2.08t ha' to 2.19t ha"' when 20kg N ha'' was applied to chickpea. On the other hand yield with 0, 25 or 50kg P2O5 ha"' was found to be 1.99, 2.14 and 2.28t ha"' respectively. In the same year, Sharma et al. observed an increase in seed yield when 18kg N ha'' was applied. Singh et al. also in the year 1989 found that reduction in recommended fertilizer rate i.e. 18kg N ha"' and 40kg P2O5 ha"' had little effect on yield, whereas yield of rape seed and especially wheat were decreased by reducing the fertilizer rates to 25-66% of the recommended rates of 120kg N + 60kg P2O5 ha"' for wheat and 80kg N + 60kg P2O5 ha"' for rape. Thakur and Jadhav (1990) in Maharashtra applied N + P2O5 @ 12.5+25,

31 Review of Literiiture

25+50 or 37.5+75kg ha"' and obtained the seed yields of 3.23, 3.52 and 3.58t ha"' respectively when compared with 3.12t ha"' without the application of nitrogen and phosphorus. At Aligarh, Yahiya and Samiullah (1995) reported that 40kg P2O5 ha' proved to be the most effective dose for nodulation, N2 fixation, leaf area, shoot dry weight, nodule dry weight and acetylene reduction. During their observations P content of shoot and root, soluble sugar content of nodules and N uptake of shoot were also increased. At the same place, Inam et al. (1996) while working on potassium noted 50kg ha"' as the best dose for pods plant"', seed yield and the biological yield of this Corp. In another field trial Yahiya et al. (1996) further observed the increase in leaf area index, shoot dry weight, nodule number, nodule dry weight, acetylene reduction activity of nodules, shoot nitrogen accumulation, shoot and root potassium content and soluble sugar content of nodules with K supply. Application of 40kg ha' proved optimum in most of the parameters studied. Khurana and Dudeja (1996) found that high nodulating (HN) and low nodulating (LN) selections of chickpea cv. ICC-4948 and ICC-5003 remained high and low nodulating respectively at two nitrogen levels (0 and 100kg N ha"'). ICC- 4948 recorded 5.4-25% higher seed yield with an increase in nitrogen level from normal to 100 kg ha"'. However, in ICC-5003 decrease in yield was observed with increasing the nitrogen level. Yadav and Srivastava (1997) at Morena (MP) in a field trial of chickpea cv. JG-315 gave 0, 20, 40 or 60kg P2O5 ha"' with and without seed inoculation with solubilizing bacteria (PSB). Highest yield was given by 60kg P2O5 ha"'+PSP followed by 60kg P2O5 alone. Similarly, Gupta et al. (1998) while working in Madhya Pradesh in a field experiment, inoculated the seeds of chickpea cv. JG-74 with Rhizobium and Bacillus or uninoculated giving 0-40kg P2O5 ha"' as single super phosphate (SSP) or 40kg P2O5 as rock phosphate. Seed yield was more with inoculation. Application of 40kg P2O5 as SSP produced the highest mean seed yield of 1.06t ha"'. Inoculation and phosphorus application increased the N and P uptake and seed crude protein content. Also in Madhya Pradesh Patel (1998) performed a field experiment under

32 Review of Literature

irrigated conditions. The crop was given 0-60kg P2O5 ha'' and various combinations of 20k:g N, seed inoculation with Rhizobium and foHar application of 2% diammonium phosphate (DAP). Mean seed yield was highest with 60kg P2O5. The combined application of 20kg N + seed inoculation with Rhizobium + foliar application of 2% DAP gave the highest seed yield. Sonboir and Sarawagi (1998) at Raipur gave different combinations of 0-60kg P2O5 ha"', phosphate solubilizing bacteria (PSB), Rhizobium and trace elements. Highest seed yield was found under the treatment 60kg P2O5 + PSB + Rhizobium + seed application of Mo and Fe. This treatment also gave the maximum nodulation. Again Sarawagi et al. (1999) reported that N and P uptake increased with increase in levels of phosphorus and was further increased with the application of phosphate solubilizing bacteria (PSB) alone or in combination with Rhizobium culture (RC). Seed yield was increased by the use of PSB and RC alongwith phosphate fertilizers. Guhey et al. (2000) also from Raipur reported an increase in seed protein content with increase in phosphorus levels i.e. 20, 40, 60 or 80kg ha''. Sugar content increased upto pod filling, then declined. Joseph and Sawarkar (1999) at Jabalpur utilized the low grade rock phosphate (RP at 80 and 160kg ha'') in meeting the phosphorus requirement by amending with farmyard manure (FYM, at 5t ha''), pyrites (at 40kg ha'') and phosphate solubilizing bacteria (PSB) and found an increase in biomass with increasing level of phosphorus. Application of RP amended with FYM resulted in the highest concentration of'P' and 'Zn' in biomass of chickpea. Singh el al. (1999) at Bilaspur (MP) observed that application of poultry manure and S. roslrata produced higher yield of rice and chickpea at 80 (N), 50 (P2O5), 30 (K2O) kg ha'' level of chemical fertilizer. Jain et al. (1999) also in MP reported that when seeds of chickpea were inoculated with Rhizobium and/or phosphorus solubilizing bacteria (PSB) and given 30, 45 or 60kg P2O5 ha'' then nodulation, pods plant'', seed and stover yield were increased with phosphorus application. Combined inoculation of Rhizobium and PSB + 60kg P2O5 produced the highest mean seed yield and the maximum net returns. Braga and Vieira (1998) at Coimbra (Brazil) in a field experiment inoculated seeds of chickpea with Bradyrhizobium or not inoculated and given 0 or 30kg N ha', 0 or 40g Mo ha'' and 0 or 40 kg micronutrients ha'' i.e. Zn, B, Cu, Fe, Mn and Mo.

33 M)SSS3» Review of Literature

Inoculation gave the maximum seed yield, followed by N fertilizer. Carrasco (1998) in Spain inoculated the seeds of chickpea with 2 strains of Rhizobium or not inoculated and applied the recommended nitrogen rate or no nitrogen. Seeds yields were highest with seed inoculation. Takankhar et al. (1998) in Maharashtra, inoculated the seed of chickpea with Rhizobium or not inoculated, and gave 0-75kg P2O5 and 25 or 50kg N ha''. Seed inoculation and the application of nitrogen and phosphorus significantly increased P uptake. The application of 75kg P2O5 produced the highest seed yield. Bhuiyan et al. (1999) at Rangpur (Bangladesh) reported that seed yield, nodulation, nodule weight shoot weight and stover yield of chickpea were highest in treatments with Rhizobium + P + K + Mo + B. The seed yield in this treatment was increased upto 204% over the unfertilized, uninoculated control. During the same year Das et al. in Himachal Pradesh found that P @ 80kg ha'' produced the highest N, P and K contents in the grain and straw, total N, P and K uptake, and grain and straw yield. Hadi and Sheikh (1999) at ElRwakeeb (Sudan) studied the effect of seed inoculation with Rhizobium and nitrogen fertilizer application @, 50kg ha''. They found that Rhizobium inoculation or nitrogen fertilizer application significantly increased the total nodule number plant'', 100 seed weight, yield and protein content of seeds. Alloush et al. (2000) at Beaver (USA) observed that when chickpea was given phosphorus in various combinations, an increase in shoot dry matter, and accumulation of P, S, Mg, Ca and K was observed. Similarly during their study on phosphorus. Mukherjee and Rai (2000) at New Delhi, observed that 0 and 60kg ha' exhibited perceptible influence on yield of the crops. Biofertilizer and phosphorus interaction showed significant influence on growth and P uptake by wheat and chickpea compared with either of the components applied separately. Krishna et al. (2001) at Kanpur studied the effect of 3 levels (0, 15 and 30kg ha"') of nitrogen on nodulation, yield and N uptake. Nitrogen at 30kg ha'' produced the highest nodule number, seed yield plant'', test weight and N uptake. At Cairo (Egypt) Shetaia and Soheir (2001) while working on yield and its components in

34 Review of Literature response to phosphoms fertilization reported that P2O5 at 40kg feddon"- I significantly increased the pods plant'', pod weight plant"', seed yield plant'' and seed and straw yield feddon''. Sawires (2001) at Giza (Egypt) found that 23.25kg P2O5 feddon' recorded the highest number of pods plant'', pod weight plant"' and seed and straw yield. Kurdali et al. (2002) at Damascus (Syria) evaluated the impact of three rates of potassium (0, 75 and 150kg K2O ha'') on nodulation, dry matter production and nitrogen fixation by faba-bean and chickpea in a pot experiment. The higher level of potassium increased both dry matter production and total N2 fixed in faba bean but did not have any effect on chickpea. Kumar et al. (2003) at Hisar reported the effect of P and K fertilizers, alone and in combination on chickpea under moisture deficit condition. Phosphorus and potassium were applied through single super phosphate and muriate of potash at the rate of 50kg ha"' each after germination. Treatment with phosphorus and potassium increased the dry weight of leaves and stem. The relative water content of leaves also increased significantly with fertilizer application. Application of fertilizers proved beneficial in terms of grain yield. Tomar et al. (2002) at Junagadh noted the effect of graded doses of potassium (0, 25, 50, 75 and 100kg ha'') on two varieties of chickpea. Its application increased the grain and straw yield, 100 seed weight, protein content and concentration of K and N in grain and straw. 50kg K2O ha'' was found optimum. Conclusion It may be concluded that wastewater has been tested under various conditions with different crops. Response to wastewater varied from region to region, crop to crop and source to source. In most of the cases lower concentration proved beneficial as it contains some essential elements while the higher was deleterious as it becomes toxic. Fly ash is another potential source of many macro- and micro-nutrients to plants although it contain some toxic metals also. Its application with various organic amendments and biofertilizer treatments can improve soil quality and lead to higher fertility. Various categories of plants can be grown on fly ash, but members of leguminosae seems to be advantageous. Plants, such as chickpea, which has a Rhizobium symbiont for nitrogen input and that exerts a metal detoxification

35 Re^'ic^v of Liteiatuie mechanism such as PC synthesis, demostrate the possibiHty to grow such crops in fly ash contaminated soil and the water incidently which is generated in large quantities from the same thermal power plant may also be used as a source of irrigation.

36 ^^Materials and ^Methods CONTENTS

MATERIALS AND METHODS

Page no.

3.1 Agro-climatic conditions of Aligarh 3 7 3.2 Preparation of pots 37 3.3 Seed treatment 38 3.4 Experiments 38 3.4.1 1 38 3.4.2 II 39 3.4.3 III 39 3.4.4 IV 39 3.4.5 V 40 3.5 Statistical analysis 40 3.6 Soil and fly ash analysis 40 3.7 Water sampling and analysis 45 3.8 Biometric observations 51 3.9 Physiological parameters 52 3.10 Yield characteristics 56 3.11 Seed analysis 56 Chapter-3 MATERIALS AND METHODS

To achieve the aims and objectives pointed out in Chapter 1 five pot experiments were performed in the net house of Environmental Plant Physiology, Department of Botany, Aligarh Muslim University, Aligarh, during the rabi (winter) seasons of 1999-2002. These studies were conducted to assess the suitability of thermal power plant wastewater (TPPW) and fly ash (FA) as potential sources of plant nutrients and irrigation water on two varieties BG-256 and Avarodhi of chickpea (Cicer arietinum L.). These treatments were also supplemented with different doses of NPK fertilizer to assess if some inorganic fertilizers could be saved keeping the economic and environmental factors in mind. 3.1 Agro-climatic conditions of Aligarh Aligarh is an industrial city famous for its locks and electroplating. It has an area of 5,024 sq kms and situated at 27°52'N latitude, 78°51'E longitude and an altitude of 187.45 MSL (Fig. 1). The climate is semiarid and subtropical, with hot dry summer and cold winter. The winter ranges from October to the end of March and the mean temperature for December and January, the two coldest months varies between 10°C-15°C. The summer extends from April to June and the average temperature for the summer remains between 32°C-35°C, whereas the maximum temperature may go even up to 46°C-47°C (Fig. 2). The average annual rainfall remains around 600- 650mm. More than 85% of the rainfall occurs during June to September and rest in winter. The soil of Aligarh varies form sandy, loamy, sandy loam and clayey loam, however, the soil used for these experiments was sandy loam. 3.2 Preparation of pots Clean earthen pots of 12" diameter with a hole at bottom for aeration were chosen and prior to filling, the farm soil was mixed thoroughly and made the total soil and fly ash 7 kg pot"' (Table la). Before sowing, in each pot, sufficient quantity of ground water together with NPK doses (Tables la,b«&c) were given 24 hrs in advance to provide necessary moisture for germination and to avoid seed injury due to fertilizers. ILEACHATE {

SAMPLINGN>V^ / SITE (WASTE WATER)N>^^y

28* 0'

^ ALIGARH ol4Km

RAILWAY STATION HARDUAGANG

ALIGARH 12 Km

Fig. 1 Map showing the location of thermal power plant, leachate reservoir and sampling site c M D

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OH i V3 B m 3 B m a 5 c m^mmm^^sms^mm o

-^.•••••• SS&^W:::¥ft¥ftWSSJ&i¥S¥^^ > o u. C3 u. lU

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iitiii»iiii'ii»ii'ii>t*iiii»iiiiin»iiiiiii»»fiiii •••'»< » fT^'i miiiiii'»t»i»i c SSS?AWft¥SftSWSiS? o

^ ^^

^

1 o ri ^ >r> •* Tf r^ f^ iaLVHHcDN31 Table la. Scheme of treatments given in Experiments I and II (Experimental design; randomized complete block design).

Irrigation Treatments water Rpmarks (g pof') GW WW FAo + + Soil FAio(600) + + Fly ash 10% FA20 (1200) + + Fly ash 20% FA40 (2400) + + Fly ash 40%

N.B. A uniform basal dose of nitrogen, phosphorus and *potassium at the rate of 20kg ha*' each was applied.

Table lb. Scheme of treatments given in Experiments III and IV (Experimental design; randomized complete block design).

Treatments Irrigation water _. , / *-i\ ^„, ;rT7^77 Remarks (gpot ) GW WW FAioNo + + Fly ash 10% and 0kg N/ha FAioNio (0.066) + + Fly ash 10% and 10kg N/ha FA10N20 (0.132) + + Fly ash 10% and 20kg N/ha FA10N30 (0.198) + + Fly ash 10% and 30kg N/ha N.B. A uniform basal dose of phosphorus and *potassium at the rate of 20kg ha'' each was appUed.

Table Ic. Scheme of treatments given in Experiment V (Experimental design; randomized complete block design).

Treatments Irrigation water (gpof') GW WW Remarks FAioPo + + Fly ash 10% and 0kg P/ha FAioPio (0.171) + + Fly ash 10% and 10kg P/ha FA,oP2o (0.342) + + Fly ash 10% and 20kg P/ha FAioPao (0.513) + + Fly ash 10% and 30kg P/ha FA,oP4o (0.684) + + Fly ash 10% and 40kg P/ha

N.B. A uniform basal dose of nitrogen and *potassium at the rate of 10 and 20kg ha' respectively was applied. * Potassium (0.128 g pof') Materials and Methods

3.3 Seeds treatment Seeds were procured from Indian Agricultural Researcli Institute (lARI), New Delhi and Agricultural Directorate, Aligarh. Authentic and viable Rhizobium culture {Rhizobium sp) specific for chickpea was also obtained from lARI, New Delhi. Healthy seeds were surface sterilized with absolute alcohol and dried in shade before applying the inoculum according to the method of Subba Row (1972). For this, 200g colourless gum Arabic (coating material) and 50g sugar were dissolved in 500ml warm water. The solution was allowed to cool and lOOg Rhizobium culture was properly mixed. Required quantity of seeds were mixed with inoculum until the seeds were evenly covered by the inoculation mixture. These inoculated seeds were spread in a clean tray to let the coating get dried in shade. 3.4 Experiments All the pot experiments were conducted according to the randomized complete block design in accordance with the scheme of treatments (Tables la,b&c). Experiments I and II were conducted in winter season (November to March) during the year 1999-2000, III and IV in the year 2000-2001, while Experiment V in 2001- 2002. 3.4.1 Experiment I It was conducted on chickpea cv. BG-256, a newly released variety from lARI, New Delhi, to compare the effect of thermal power plant wastewater (TPPW) and ground water (GW) alongwith fly ash. TPPW was collected from the outlet of the leachate reservoir (Fig. 1) of Harduaganj Thermal Power Plant, Kasimpur, located 13km away from Aligarh city, whereas tap water, without any treatment, was the source of GW. Fly ash was also collected from the fly ash pond of the same thermal power plant. Each pot received 500ml water on ahernate days for the duration of about 125 days starting from 10*'' day after sowing (DAS) i.e. after seedling emergence. Four different concentrations of fly ash as 0, 10, 20 and 40% were thoroughly mixed with soil making the total of soil/fly ash weight upto 7kg pot"' (Table la). The control consists of only soil without fly ash. Uniform starter basal dose of nitrogen (20kg ha"'), phosphorus (20kg ha"') and potassium (20kg ha"') was also applied before sowing (Tables la,b&c). The sources of NPK were , single

38 Mutcriiils and Methods

super phosphate (SSP) and muriate of potash (MoP) respectively. Sowing was done on T^ November 1999, at the rate of 10 seeds pot"' to avoid germination failure and harvesting on SO**" March 2000. Weeding was done whenever weeds emerged. For each treatment seven pots containing two plants each, keeping a uniform distance, after thinning were maintained. Three plants were collected randomly for each sampling at vegetative, flowering and fruiting stages and remaining five plants were considered for the study of yield and its characteristics. 3.4.2 Experiment II This experiment was conducted simuhaneously with Experiment I on another cultivar of chickpea i.e. Avarodhi, a local and commonly grown variety to check, which one of the two varieties performed well grown under similar treatments. All the cultural practices like thinning, weeding, sampling and harvesting were same as explained earlier. Sowing was done on S'"" November, 1999 and harvesting on 31'' March, 2000. 3.4.3 Experiment III It was conducted during the following year on chickpea cultivar BG-256, to strengthen the findings of earlier experiment with inorganic fertilizer doses. Here again the comparative effect of TPPW and GW was studied. On the basis of observations made in Experiment I, the best concentration of fly ash i.e. 10% was selected and added to the soil, making the final weight of fly ash amended soil upto 7kg ha''. Different doses of nitrogen i.e. 0, 10, 20 and 30kg ha'' were supplemented in order to workout the optimum dose for cultivar BG-256. A uniform basal dose of phosphorus and potassium at the rate of 20kg ha'' each was also applied before sowing (Tables Ib&c). The sources of NPK were the same as in previous experiments. Healthy seeds of more or less uniform size were surface sterilized and then inoculated. The seeds were sown on 2"'' November 2000 and harvested on 30"' March 2001. Seven pots containing two plants each were maintained for each treatment. All other cultural practices were the same as undertaken earlier. 3.4.4 Experiment IV This experiment was carried out together with Experiment III on cultivar, i.e. Avarodhi to compare the performance of this variety against BG-256 grown under

39 Materials and Methods

TPPW, GW and varying doses of nitrogen i.e. 0, 10, 20 and 30kg ha"'. The phosphorus and potassium doses were uniform as in Experiment III to workout the optimum concentration of nitrogen. The sources of NPK and all other cultural practices were also same as in Experiment III. The seeds were sown on 2" November, 2000 and harvested on 30*^ March, 2001. 3.4.5 Experiment V This experiment was conducted during the year 2001-2002, based on the findings of experiments I-IV. The aim of this experiment was to evaluate the optimum phosphorus dose against 10% fly ash-soil amendment as well as source of irrigation water (Table Ic). In this experiment, chickpea cultivar BG-256 was selected on the basis of its better performance in earlier experiments. A uniform basal dose of nitrogen at the rate of 10kg ha'' based on the findings of earlier experiments III and IV, was also applied along with 20kg K ha"'. The sources of fly ash, irrigation water, NPK fertilizer and other cultural practices were the same as explained earlier. The seeds were sown on 3"* November, 2001 and harvested on 31^ March, 2002. 3.5 Statistical analysis The data for the growth and yield of each experiment were analysed statistically taking into consideration the variables according to Panse and Sukhatme (1985). The 'F' test was applied to assess the significance of data at 5% level of probability (p<0.05). The error due to replication was also determined. The model of analysis of variance (ANOVA) is given in Tables 2a&b. Critical difference (CD.) was calculated to compare the mean values of various treatments. In addition, the pooled analysis of Experiment (I and II) and (III and IV) was also worked out according to split-split plot design (Table 2c). Correlation coefficient values (Table 66) and regression (Figs. 17-21&24-32) of seed yield with some growth, physiological and yield attributing parameters were also undertaken. 3.6 Soil and fly ash analysis The fly ash was collected form the ash ponds dumped at 'Harduaganj Thermal Power Plant' (HTPP), Kasimpur, located 13 kilometers away from Aligarh city. Before mixing the soil and fly ash, samples of the two were taken. Small quantity of the soil and fly ash were ground separately with the help of mortar and pestle and

40 Table 2a. Model of analysis of variance (ANOVA) of Experiment I and IV (Experimental design; randomized complete block design)

Source of variation df SS MSS F. value Sig. Replication 2 Water I Fly ash/Nitrogen 3 Interaction 3 Error 14 Total 23

Table 2h. Model of analysis of variance (ANOVA). Pooled analysis of Experiment I & II and III &. IV (split-split plot design)

Source of variation df SS MSS F. value Sig. Replication 2 Variety (A) 1 Error (a) 2 Water (B) 1 AxB 1 Error (b) 4 Fly ash/Nitrogen (C) 3 AxC 3 B xC 3 AxB X C 3 Error (c) 24 Total 47

Table 2c. Model of analysis of variance (ANOVA) of Experiment V (Experimental design; randomized complete block design)

Source of variation df SS MSS F. value Sig. Replication 2 Water 1 Phosphorus 4 Interaction 4 Error 18 Total 29 Materials and Metliods passed through a 2mm sieve. The following chemical characteristics were studied in soil as well fly ash separately (Tables 3&4). 1. Hydrogen concentration (pH) It was estimated with the help of pH meter. To lOg soil/fly ash, 50ml double distilled water (DDW) was added and shaken thoroughly. After 30 minutes, pH of the suspension was recorded. The pH meter was calibrated with a standard buffer of known pH (Jackson, 1973). 2. Cation exchange capacity (CEC) CEC of both samples was determined by the method of Ganguly (1951). To lOg soil/fly ash, 0.2N HCl (Appendix, p.ii) was added. It was shaken for 30 minutes, filtered and washed with DDW, till it became free from chloride , which was checked with AgNOs. The residue was transferred from the filter paper to a beaker and suspension of known concentration was prepared. It was then treated with 10ml of standard KCl solution, shaken for 30 minutes and left overnight. It was titrated with 0. IN NaOH (Appendix, p.iii), using phenolphthalein (Appendix, p.iii) as an indicator. From the amount of NaOH required, the CEC of the sajnples was calculated as follows volume of 0. IN NaOH x N of NaOH CEC (meq lOOg-') = weight of the sample

3. Organic carbon It was estimated by the method of Walkley and Black (1934). 2g sample was taken in a 500ml conical flask. To this lOml of IN potassium dichromate solution (Appendix, p.iii) and 20ml concentrated sulphuric acid were added. After shaking for about 2 minutes, it was kept on an asbestos mat for 20 minutes. 200ml DDW, 10ml (85%) and 1ml of diphenylamine indica .>r (Appendix, pi) were added. Deep violet colour appeared which was titrated with 0.5N ferrous sulphate solution (Appendix, p.i) till the colour changed to purple and finally green. Simultaneously a blank was run without sample. blank titre - actual titre % of organic carbon = x 0.003 x 100 x N weight of sample

4! o o ON vo ON ?; 2 o o O o > 00 00 00 I/O oj o K r^ 00 o o o ON

•a

'o (U

CO c<3 00 o t^ vo o ON CNJ O o vo —' o rn vo en ro in ^ (S 00 (S •— o 1-H ON rn ON iri r^' vd fn oo'

l-l a B l-l 'C u tl-l e-. ro (S •—' 00 00 •o 00 00 ^ o m 00 -—I 00 ts o o o r>4 m •—•

60 e •S C o O c O VO t^ •^ CO ON VO M- o 2 o 00 CO O — —' o '=> ^ ^ ^ ON 00 vo CN iri r~ m 00

60 c

o o o r- ON o t^ vo ^- to CO oo vo 00 *—t o o ro VO CO •"5- vd fS O vo CN ^^ vd c^' 00 .—• ON vd o o o 1—< CO »—< VO o CO O CO O

a; o a) Ui o ea o y~\ /^ CO to o^ ^ JT^ o v^—^ 'M 60 J>S o O c •g c O o o 'eo CO 'B X) CO O -•-» cr c<3 E cx O ON m

T3 (L> tH O /^ v^ 1—I O O in 00 NO f*^ I—< NO c~ CoO ON 00 ^ 2 "^ <^ VO Ui +-• C/} x •*-» oUi C O 0) .E 1 CL O X \ • W o »o o CO 0\ NO TT O o\ o CNl o CN| CS CN »—< ro ON 1—1 r- CS o »n NO •^ NO ON t~- 00 CS CO '" o\ m o (S NO CN

B a v> C ,o

o O VO 00 c VO rs V-) o r- o q CNl CS 1—< NO o rs 'E O 00 U-) •<3- 00 00 o ON m 00 i_ CS Os O ~ o VO CS •4-J •a

«50 c •o| «5 O CO o c> VO C«l o o CS o rs o CS 00 r~^ c2 NO C3N oo' m m CS VO ON '^. '^n CD CS CS m CN — ON ON to

W5

o J3 00 to o 'on o "cin o c E C o o o "E X) o E u 2 <0 cr £ 3 to a 4> c E oc 4-* o E c n CO •a c 3. on

where, N - normality of ferrous ammonium sulphate. 4. Nitrate nitrogen It was estimated according to the method of Ghosh e( al. (1983). 20g soil/fly ash samples were shaken continuously with 50ml DDW for 1 hour in a 100ml conical flask fitted with rubber stopper. A pinch of CaS04 was added and shaken. Then the contents were filtered through a Whatman No. 1 filter paper. 20ml clear filterate was transferred to 50ml porcelain dish and was evaporated to dryness on steam bath. After cooling, 3ml phenol disulphonic acid (Appendix, p.ii) was added and allowed to react for 10 minutes. 15ml DDW was added and stirred with glass rod until the residue was dissolved. After cooling, the contents were washed down into 100ml volumetric flask, to this 1:1 liquid (Appendix, p.ii) was added slowly and mixed well, till the solution became alkaline which was indicated by the appearance of yellow colour due to the presence of nitrate. Then, another 2ml ammonia was added and finally the volume was made upto 100ml with DDW. The intensity of the yellow colour was read at 410nm with spectrophotometer. For standard curve, stock solution containing 100 ppm nitrate was prepared by dissolving 0.722g potassium nitrate in DDW and the volume was made upto 1 litre. This was diluted ten times to give 10 ppm NO3-N soluUon. Aliquots (2, 5, 10, 15, 20 and 25ml) were evaporated on water bath to dryness in small porcelain dishes. After cooling, 3ml phenol disulphonic acid was added and yellow colour was read as described above. Simultaneously, a blank was also run. 5. Phosphorus To 2.5g soil/fly ash sample in 100ml conical flask, a pinch of Draco GGO was added followed by 50ml of Olsen's reagent (Appendix, p.ii). The flask was shaken for 30 minutes on a shaker and then the contents were fUtered through a Whatman No. 1 filter paper. In the fiherate, phosphorus was estimated through spectrophotometer by Dickman and Bray's (1940) method. 5ml soil/fly ash extract was pipetted into a 25ml volumetric flask and 5ml Dickman and Bray's reagent (Appendix, p.i) was poured drop by drop with constant shaking fill effervescence due to CO2 evolution ceased. The inner wall of the flask

42 Materials and Methods neck was washed with DDW and the contents diluted to 22ml. Then 1ml stannous chloride solution (Appendix, p.iii) was added and the volume was made upto the mark. The intensity of blue colour was read at 660nm on spectrophotometer. For the standard curve, 0.43 9g potassium dihydrogen orthophosphate (KH2PO4) was dissolved in about half litre of DDW. To this, 25ml 7N sulphuric acid (Appendix, p.iv) was added and volume was made upto 1 litre with DDW, giving 100 ppm stock solution of P. From this, 2 ppm solution was made after 50 times dilution. For the preparation of the standard curve, different concentrations of? (1, 2, 3, 4, 5 and 10ml of 2ppm phosphorus solution) were taken in 25ml volumetric flasks. To these, 5ml of extracting reagent (Olsen's reagent) was added. The colour was developed by adding Dickman and Bray's reagent and stannous chloride and read at 660nm. A blank was also run without the sample. The curve was plotted and the amount of P was calculated from the curve. 6. Potassium 5g soil/fly ash was shaken with 25ml IN (Appendix, p.i) for 5 minutes and was filtered immediately through a Whatman No. 1 filter paper. Stock solution of 1000 ppm K was prepared by dissolving 1.908g KCl in 1 litre DDW. From the stock solution aliquots were diluted in 50ml volumetric flask with ammonium acetate solution to give 10 to 40 ppm of K. These were read with the help of flame photometer after setting zero for the blank at 100 for 40 ppm of K. The curve was obtained by plotting the readings against the different concentrations (10, 15, 20, 25, 30, 35 and 40 ppm) of K. 7. Sodium Determination of Na was carried out directly from the soil/fly ash extract (1:5, soil/fly ash : water) with the help of flame photometer. Standard curve was prepared by taking 5.845g NaCl dissolved in DDW and the volume was made upto 1 litre which gave 100 milli equivalents litre'' of Na. From this stock solution, dilutions containing 5, 10, 15, 20, 25, 30, 35, 40, 45 and 50 meq Na litre"' were prepared. The curve was drawn by plotting the flame photometer readings on Y-axis against concentrations of Na on X-axis. Na in the unknown sample was read from curve.

43 Materials and Methods

Preparation of extracts for calcium, mngnesium, chloride, carbonate-bicarbonate and sulphate lOOg sample transferred to 750ml flask, to this 500ml DDW was added and the flask was shaken for about 1 hour. The contents were filtered through Buchner funnel. 8. Calcium It was estimated according to the method of Chopra and Kanwar (1982). To 25ml extract, 2-3 crystals of carbamate and 5ml of 16% NaOH solution were added. Then it was titrated with 0.0 IM EDTA (Appendix, p.i), using indicator (Appendix, p.ii) till the colour changed from orange red to purple. 9. Magnesium To 25ml sample extract, 5ml ammonium chloride-ammonium hydroxide buffer (Appendix, p.i) was added, followed by titration with 0.0IM EDTA (Appendix, p.i), using Eriochrome Black-T (Appendix, p.i) as an indicator, the colour changed from green to wine red (Chopra and Kanwar, 1982). 10. Chloride 50ml sample extract was taken in a flask and 2ml of 5% K2Cr04 indicator (Appendix, p.iii) was added. It was titrated against 0.02N silver nitrate solution (Appendix, p.iii). (mlxN) of AgNOsx 1000x35.5 Chloride (mgl'') = ml sample

11. Carbonate and bicarbonate Estimation was done following the method of Richards (1954). 50ml soil/fly ash sample extract was taken in a clean flask. To this, 5 drops of phenolphthalein indicator were added. The appearance of pink colour indicated the presence of carbonates. Then it was titrated against 0.0IN hydrochloric acid (Appendix, p.ii) till the solution became colourless. To the colourless solution from the above titration, 2 drops of methyl orange indicator (Appendix, p.ii) were added. It was then titrated against O.OIN sulphuric acid till the colour changed from yellow to rose red. This indicated the presence of bicarbonate in the sample.

44 Materials and Methods

1000 (a) carbonate (tneq 1'') = 2Y x normality of H2SO4 x ml aliquot

= 2Yx2 1000 (b) bicarbonate (meq 1"') = (Z - 2Y) x normality of H2SO4 x ml aliquot

= (Z - 2Y) X 2 where, Y = reading of burette for the titration of carbonate Z = reading of burette for the titration of bicarbonate 12. Sulphate To 50ml sample extract, 2.5ml conditioning reagent (Appendix, p.i) was added. It was then stirred on a shaker and during shaking small quantity of barium chloride was added. It was then read with the help of nephelometer. mg SO4 X 1000 S04(mgr') = — ml sample

3.7 Water sampling and analysis For this purpose TPPW was collected twice, in the last week of March and in the first week of May in 2 litres plastic bottles from the outlet of the leachate reservoir (Fig. 1) of Thermal Power Plant while for watering of pots, it was collected in 50 litres jerry canes at weekly intervals. The source of GW was tap water. Sample bottles were carefully cleaned before use with chromic acid cleaning mixture. Later the bottles rinsed thoroughly with tap water and then with distilled water. 4 litres of water sample was taken for analysis (Table 5). Prior to filling, the sample bottles were rinsed out three times with wastewater to be collected. 1. Electrical conductivity (EC) It was directly read with the help of conductivity meter by putting the sample in a beaker. The apparatus was adjusted to 25°C of the solution. 2. Total solids (TS) 100ml unfiltered sample was taken on evaporating dish and was allowed to

45 ON •^ 0\ ON ^^ OO — . • 0\ .«• _; .^ ^; nn Ti- 00 . ^. ^. \J-,vo

00 O. B E f^ •* ^ r-~ VO ON ^^ f^ S^ - 5 -• ^>:- ^- 2 ^- - -^ ? . . • T:r cCO O o o cs

C 00 r~ ON en ON •g c.,osoNrNi;i;«N-- en ^ CM 5- s:! S S ^ NO .—1 m 0) OH oog??SSod^<^^ ^ 2 ••-» u

I ^ . ^ (N 00 VO ^ NO NO ON m t~- ^ OS 00 o m >i^ t^ (S ,— ro n ^ t--' «r) 00 >yS 00 o o cs" <^ cs OH OH

H, CS NO c^ r- NO •^ m cs _. oogg -s: CN • o^ fi m CN) r~ u 00 00 J_J NO NO ^ (S 00 OO' •^' _<• ON r^ w~, n . . vo r^ Tl- 00^ cs en O ^ '— >n

•4-* "a CO c« m ON 00 00 -e 00 Tl- NO O CO W-) o r- ^ o •* ?5. ?;; ^. --.-:'- ?i ^ ON cs 00. fi. ?;;=: r; t~- t-~ <3N vn ^ C ^^ - t-^ - rr~n 0—0 TcTs ^°°- u-—i ^ cs w-NOi o^ _o• c_s; "^

NOrJ-r-ON ElifslCTN^ 00 t^ 00 X ^ ON O ON n 00 NO cs rn 00 "T f::5 2< j^ i/^ 00 cs ON C^ NO o f-- 00 J^j NO NO g f?, NO •—I NO Tf °N '^ ""•—^( r-1 I—1 in bO <*• ^ '^ »—« E "a E

••-» 00 - 2 S S S - -. -. - J^ - - r; - ^ ?: 2 ^ -a t-r-ONto^^oot-No^r-^rNj^^^^.^ c CTJ

0\ [;: ON r- o 2 o r- cs o 00 &; f^ Sm 0S 0 00 S iri fW 00 00 00 rn § . 00^ ^NoJ^cs-^ — '^ 2 cd E^ CQ c 00 ^. ^ ::: S ?; ^. ^. -. ^. S ^ o ON n IS ?2 5: -- 3 t~~K-c?N»?>-5i-^ooNOw-i„-NO^^oo_-_--;»^ O — r«i ^ cs "" ^- NO o o cs k> 60

00 O ^ rn j:^ CNJ r- C3N NO (S f-- t~- ON m CS O ^ .-I in c r~~ ON J_J NO NO i::j CS ^ - ^ Q. u E 00 Tt f«^ ^ k. 00 O 00 CQ v00^1"ONf«1rsirooOrN|^r<-i»^ir,/-.Tfr~. «/0 p-l ^ (s) 1 . _. _. \1 m ". CS 00 ^. f^ 0—0 m-- O_N. u 00 r-- ON »n •rt- r>- o 00 NO ^ NO ^ - pi ~ -"i- ^ cs O O CS

u 'E in 'o u

O CO

z z 00 00 Q Q K U 00 Q 00 O O 00 a y o ^ o o H Q,WHh-HCQOStjy:2EUO&.Z o C/2 MiUciiiils and Methods evaporate on water bath. A-B X 1000 Total solids (g 1'^) = V

Where, A = Final weight of the dish B = Initial weight of the dish V = Volume of the sample taken 3. Total dissolved solids (TDS) 100ml filtered sample was taken in an evaporating dish and allowed to evaporate on water bath. A-B X 1000 TDS (g r') = V

Where, A = Final weight of the dish B = Initial weight of the dish V = Volume of the sample taken 4. Total suspended solids (TSS) These were determined by calculating the difference between the total solids and total dissolved solids. TSS (g r') = TS - TDS 5. Hydrogen ion concentration (pH) It was determined with the help of pH meter. The pH meter was adjusted before use with standard buffer of known pH. 6. Dissolved oxygen (DO)/Biological oxygen demand (BOD) Different volumes of the samples were placed in BOD bottles (250ml) to get several dilutions of the samples to obtain the required depletions ranging between 0.1 and 1.0%. These bottles were filled with DDW, stoppered and one set of bottles was incubated for 5 days in an incubator maintained at 20°C and in other set, dissolved oxygen (DO) was determined immediately by adding 2ml manganous sulphate solution (Appendix, p.ii), followed by 2ml alkali iodide azide reagent (Appendix, pi), by means of graduated pipette by dipping its end well below the surface of the liquid.

46 Materials and Methods

The BOD bottles were stoppered and mixed well by inverting. The bottles were allowed to stand till the precipitate settled half way, leaving clear supernatant above the manganese hydroxide flakes. The stopper was removed and 2ml H2SO4 was immediately added. Each bottle was restoppered and the contents were mixed by gentle inversion until dissolution was complete. 200ml sample was taken in 500ml conical flask, then 2ml starch indicator (Appendix, p.iii) was added and titrated against 0.025N sodium thiosulphate solution (Appendix, p.iv) till the disappearance of blue colour. The reading of the sodium thiosulphate used up was indicative of DO of the sample in mg 1*'. BOD was calculated using the following relationship D,-D2 BOD (mg r') = P where, Di and D2 are the DO of the diluted samples, 15 minutes after the preparation of the sample and after 5 days of incubation respectively and P is the decimal fraction of the sample used. 7. Chemical oxygen demand (COD) 0.4g mercuric sulphate was placed in a refluxing flask and 20ml sample was added. Both were mixed well and 10ml 0.25N potassium dichromate solution (Appendix, p.iii) was added followed by 30ml sulphuric acid and small amount of silver sulphate. A blank was run using distilled water. These were subjected to reflux for two hours, cooled and then diluted to about 100ml DDW. The contents were titrated against O.IN ferrous ammonium sulphate solution (Appendix, p.i). A - B X N X 8,000 COD (Mg 1') = ml sample where, A = ml ferrous ammonium sulphate used for blank titration B = ml ferrous ammonium sulphate used for sample titration N = normality of ferrous ammonium sulphate solution 8. Calcium 50ml ground water/wastewater sample was taken in conical flask and neutralized with acid. It was boiled for 1 minute and then cooled. 2ml IN sodium

47 Materials and Methods hydroxide solution (Appendix, p.iii) added to maintain the pH at 12-13. After the addition of 1-2 drops of ammonium purpurate indicator (Appendix, pi), it was titrated slowly with 0.0IM, EDTA and calculated as follows, A X B X 400.8 Ca (mg r') = ml sample where, A = ml titration for sample B = mg CaCOs equivalent to 1.0ml EDTA titrant at the calcium indicator end point 9. Total hardness 50ml ground water/wastewater sample was taken in conical flask and 1ml ammonium chloride-ammonium hydroxide buffer solution was added. After the addition of lOOmg Erichrome Black T indicator it was titrated against O.OIM, EDTA solution. ml EDTA used X 1000 Hardness as mg 1"' CaCOs = ml sample

10. Magnesium It was estimated from EDTA and hardness titration (taken from total hardness estimation). Mg (mg J'') = Total hardness (as mg CaCOa 1'') - calcium hardness x 0.244 (as mg CaCOs l') 11. Chloride 50ml sample was taken in a flask and 2ml potassium chromate indicator was added. It was titrated against 0.02N silver nitrate solution. (mlxN) of AgNOaX 1000x35.5 Chloride (mg I"') = ml sample 12. Potassium The estimation of potassium was carried out directly with flame photometer at 768nm using appropriate filter and a standard curve by taking concentrations of potassium. A stock solution of 1,000 ppm K was prepared by dissolving 1.908g KCl in 1 litre DDW.

48 Materials and Metiuid.s

13. Sodium Sodium was also estimated flame photometrically at 589nm using the specified filter and standard curve by taking known concentrations of sodium salt. For standard curve 5.845g sodium chloride was dissolved in DDW and the volume maintained at 1 litre. This gave 100 meq I"' of Na. From this stock solution, dilutions containing 5, 10, 15, 20, 25, 30, 35, 40, 45 and 50 meq Na 1'" were prepared. A curve was drawn by plotting the flame photometer readings on Y-axis against concentrations of sodium on X-axis. The concentration of sodium in the unknown sample was read from the curve. 14. Phosphate To 100ml sample containing not more than 0.2mg phosphorus and free from colour and turbidity, 0.05ml phenolphthalein indicator was added. Sample turned pink, strong acid solution was added drop-wise to discharge the colour. Smaller sample was taken and diluted to 1,000ml with DDW. After discharge the pink colour with acid, 4ml ammonium molybdate reagent (Appendix, p.i) was added. After 10 minutes the colour was measured with the help of spectrophotometer at 690nm and comparison with the calibration curve was made, using DDW blank. mgP X 1,000 P(mgr') = - ml sample

15. Sulphate In a conical flask 100ml sample was taken and 5ml conditioning reagent was mixed. The contents of the flask were stirred for one minute on magnetic stirrer and small amount of barium chloride was added during stirring. It was then read at 420nm with the help of spectrophotometer. Standard sulphate solution was prepared by dissolving 0.1479g sodium sulphate in DDW, making the volume 1 litre. From this 0- 40mg r' dilutions were prepared at the interval of 5mg l'. A standard curve was prepared by plotting the readings for each duration using spectrophotometer. 16. Nitrate nitrogen First nitrate standard was prepared in the range of 0.1 to l.Omg p' N by diluting I, 2, 4, 7 and 10ml standard nitrate solution to 10ml with DDW. Residual

49 Materials and Methods chlorine in the sample was removed by adding 1 drop sodium arsenite solution for each 0. lOnig CI and mixed. One drop was added in excess to 50ml portion. For colour development, numbers of reaction tubes were set in wire rack. To each tube 10ml sample was added. The rack was placed in cool water bath and 2ml NaCl solution was mixed well. Then 10ml H2SO4 was added and cooled. 0.5ml sulphanilic acid solution (Appendix, p.iv) was added and the tubes swirled to mix and then placed in water bath at not less than 95°C. After 20 minutes, it was taken out and cooled in a cold water bath. Reading was taken against a reagent blank at 410nm. Standard curve was prepared from the absorbance values of the standard run together with the samples and correlated by subtracting their 'sample blank' values from their final absorbance values. The concentration of NO3-N was read directly from the standard curve. 17. Ammonia nitrogen For the estimation of ammonia nitrogen, first preliminary distillation was performed. 500ml ammonia free water was added to 20ml borate buffer and the pH was adjusted to 9.5 with 6N sodium hydroxide solution (Appendix, p.iii). A few glass beads were added and the mixture was used to steam out the distillation apparatus until the distillate showed no traces of ammonia. For ammonia nitrogen content of less than lOO^g 1'', volume of 4 litres was used. Residual chlorine was removed in the sample by adding dechlorinating agent, 25ml borate buffer was added and the pH was adjusted to 9.5 with 6N NaOH, using pH meter. Distillation of sample was done. The steaming out flask was disconnected and the sample was immediately transferred to the distillation apparatus. It was distilled at the rate of 6 to 10ml minute'' with the tip of delivering tube submerged. The distillate was collected in 500ml flask, containing 50ml boric acid solution. At least, 300ml distillate was collected. It was diluted to 500ml with ammonia free water. 100ml sample was taken in 500ml kjeldhal flask with ammonia free distilled water and diluted to 250ml. Again, it was distilled as before with few pieces of paraffin added to the distillation flask and 100ml distillate was collected. Ammonia in the distillate was titrated against standard 0.02N H2SO4 titrant (Appendix, p.iv) until the indicator turned to pale lavender. A blank was run through all the steps of the procedure.

50 Materials and Methods

(A - B) X 280 Ammonia N (mg 1'') = ml of sample where, A = ml H2SO4 titration for sample B = ml H2SO4 titration for blank 18. Carbonate and bicarbonate Estimation was done following the method of Richards (1954). 50ml water sample was taken in a clear flask. To this, 5 drops of phenolphthalein indicator were added. The appearance of pink colour indicated the presence of carbonate. Then, it was titrated against COIN sulphuric acid till the solution turned colourless. To the above solution, 2 drops of methyl orange indicator were added. It was again titrated against 0.0IN H2SO4 till the colour changed from yellow to rose red. This indicated the bicarbonate presence. 1000 (a) carbonate (meq 1'') = 2Y x normality of H2SO4 x ml aliquot

= 2Yx2 1000 (b) bicarbonate (meq 1"') = (Z - 2Y) x normality of H2SO4 x ml aliquot where, Y = reading of burette for the titration of carbonate Z = reading of burette for the titration of bicarbonate 3.8 Biometric observations For investigating the comparative effect of TPPW, GW and fly ash under inoculated conditions, observations were carried out at vegetative, flowering, fruiting and at harvest stages. For the study of the root, the plants were uprooted carefully and washed gently to clear all the adhering particles. 3.8.1 Growth characteristics The following growth characteristics were observed using standard methods 1. Shoot length (cm plant"') 2. Shoot fresh weight (g plant'')

51 Materials and Methods

3. Shoot dry weight (g plant'') 4. Leaf nu mb er p lant'' 5. Leaf area (cm^ plant'') 6. Branch number plant'' 7. Root length (cm plant'') 8. Root fresh weight (g plant"') 9. Root dry weight (g plant"') 10. Nodule number plant"' 11. Nodule fresh weight (g plant"') 12. Nodule dry weight (g plant"') For assessing dry weight, three plants form each treatment were dried, after taking their fresh weight, in hot air oven at 80°C for two days and weighed. The area of leaves was measured using leaf area meter (LA 211, Systronics, India). For nodule number, whole plant was uprooted with the precaution that the roots or the nodules may not be damaged. Samples were washed gently to wipe away all the adhering foreign particles and the number was carefully counted. 3.9 Physiological parameters Following parameters were studied at vegetative, flowering and fruiting stages. L Nitrate reductase activity (NRA) NRA was estimated by the method of Jaworski (1971). Random samples of leaves from each plant were taken and cut into small pieces. 200mg fresh leaf pieces were weighed and placed in polythene vials. To each, 2.5ml phosphate buffer (0. IM) pH 7.5 (Appendix, p.iii) and 0.5ml potassium nitrate (0.2M) solution (Appendix, p.iii) were added, followed by addition 2.5ml of 5% isopropanol (Appendix, p.ii). Lastly, two drops of chloramphenicol solution were added to avoid bacterial growth in the medium. The vials were incubated for 2 hours in the dark at 30°C. 0.4ml incubated mixture was taken in a test tube to which 0.3ml of 1% sulphanilamide (Appendix, p.iv) and 0.02% N-1, napthylethylene diamine hydrochloride (NED-HCl) (Appedix, p.ii) were added. The solution was left for 20 minutes for maximum colour development. It was diluted to 5ml with DDW and optical density was read at 540nm

52 Materials and Methods

using a spectrophotometer. A blank consisting of 4.4ml DDW and 0.3ml each of sulphanilamide and NED-HCl, was used simultaneously for comparison. Standard curve was plotted by taking known graded dilutions of sodium nitrate from a standard of this salt. The optical density of the samples was compared with the calibrated curve and NRA was expressed as x] mol NO2 g''h'' fresh leaf tissue. 2. Chlorophyll estimation It was estimated following the method of Mac Kinney (1941). Fresh leaves (lOOmg) were homogenised in mortar with sufficient quantity of 80% . The extract was filtered and supernatant collected in the volumetric flask. The process was repeated thrice and each time supernatant was collected in the same flask. Finally the volume was made upto 10ml with 80% acetone. 5ml sample of chlorophyll extract was transferred to a cuvette and the absorbance was read at 645 and 663 nm on spectrophotometer. The following equation given by Arnon (1949) was adopted to calculate the total chlorophyll content. V Total Chlorophyll(mgg') = [20.2(D645) + 8.02 (D663)] x 1000 X W where, V = total volume of the solution (ml) W = weight of the tissue (g) used for the extraction of the pigments 3. Leaf analysis Healthy leaves were collected at different samplings stages for the estimation of N, P and K contents. Oven dried leaves from each treatment and replication were powdered with mortar and pestle and passed through 72mm mesh sieve. The powder was stored in small polythene bags with identification. Digestion ofleafsamples The leaf powder was oven dried before digestion and 50mg powder from each replicate was transferred to 50ml kjeldhal flask to which 2ml sulphuric acid was added. The contents of the flask were heated on temperature controlled assembly for about 2 hours to allow complete reduction of nitrates present in the plant material. As a result, the contents of the flask turned black. After cooli a the flask for about 15

53 Materials and Methods

minutes, 0.5ml 30% H2O2 was added drop by drop and the solution was heated again till the colour changed from black to light yellow. Again aftei^ cooling for 30 minutes, an additional 3 drops of 30% H2O2 were added, followed by heating for another 15 minutes. The process was repeated till the contents of the flask turned colourless. The peroxide digested material was transferred from Kjeldhal flask to 50ml volumetric flask with three washings of DDW. The volume of the flask was made upto the mark. This peroxide digested material was used for the estimation of N, P and K contents. Estimation of nitrogen Nitrogen was estimated according to Lindner (1944). 10ml aliquot of the peroxide digested material was taken in 50ml volumetric flask. To this, 2ml 2.5N sodium hydroxide (Appendix, p.iii) was added which neutralized the excess of acid and 1ml 10% sodium silicate solution was added which prevented the turbidity. The volume of the solution was made upto the mark. In 10ml graduated test tube, 5ml of this solution was taken and 0.5ml Nessler's reagent (Appendix, p.ii) was added. The final volume was made upto 10ml with DDW. The contents of the test tube were allowed to stand for 5 minutes for maximum colour development. Then the solution was transferred to colorimetric tube and optical density was read at 525nm. For standard curve of nitrogen, 50mg ammonium sulphate was dissolved in 1 litre DDW. From this solution, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 and 1.0ml was pipetted in 10 different 10ml graduated test tubes. The solution in each test tube was diluted to 5ml. In each test tube 0.5ml Nessler's reagent was added and the final volume was made upto the mark. After five minutes, the optical density was read at 525nm. A blank was run with each set of determination. Standard curve was plotted using different concentrations of ammonium sulphate solution versus optical density and with the help of this standard curve, the amount of nitrogen present in the sample was determined. Estimation of phosphorus The method of Fiske and Subba Row (1925) was adapted. 5ml aliquot was taken in lOml graduated test tube and I ml molybdic acid reagent (2.5%) (Appendix, p.ii) was carefully added, followed by the addition of 0.4ml l-amino-2 naphthol-4- sulphonic acid (Appendix, p.i). To this content volume was made upto 10ml. The

54 ^-ti v. .'

solution was shaken for 5 minutes 'for maximmin^^ c^mr development and subsequently transferred to colorimetric !ube.,T||^Qj)tjcfetathsity was read at 620nm. For standard curve of phosphorus 351mg monobasic dihydrogen orthophosphate was dissolved in sufficient DDW to which 10ml ION H2SO4 was added and the fmal volume was made to 1000ml. From this solution 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 and 1.0ml was taken in ten different graduated test tubes. The solution in each test tube was diluted to 5ml. In each tube, I ml molybdic acid reagent and 0.4ml 1-amino- 2-naphthol-4-sulphonic acid was added and the final volume was made upto 10ml. After 5 minutes, optical density was read at 620nm. A blank was run with each set of determination. Standard curve was plotted using different dilutions of potassium dihydrogen orthophosphate solution versus optical density. With the help of the standard curve, the amount of phosphorus present in the sample was determined. Estimation of potassium It was estimated with the help of flame photometer. 10ml aliquot was taken and read by using the filter for potassium. A blank was also run side by side with each set of determination. The readings were compared with calibration curve plotted using known dilutions of standard potassium chloride solution. For standard curve for potassium 1.91g potassium chloride was dissolved in 100ml DDW, of which 1ml solution was diluted to 1000ml. The resuUing solution was of 10 ppm potassium. From this 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10ml solution was transferred to 10 vials separately. The solution in each vial was diluted to 10ml. The diluted solution of each vial was run separately. A blank was also run with each set of determination. Standard curve was prepared using different dilutions of potassium chloride solution versus readings on the scale of galvanometer. The amount of potassium present in sample was determined with the help of standard curve. 3.10 Yield characteristics For this, five plants from each treatment were collected at the time of harvest, i.e. when the crop attained physiological maturity. Following yield characteristics were observed, 1. 100 seed weight 2. Podsplanf'

55 MatcriaLs and Methods

3. Seed yield 4. Biomass 5. Harvest Index 6. Seed protein content 7. Seeds pod'' Total seeds threshed out of dried plant samples of each treatment were cleaned and allowed to dry under the sun for few hours and the weight was recorded to complete the seed yield. Total biomass was recorded after drying the entire plant in the sun to make the weight constant. While for harvest index it was computed after dividing the seed yield by biomass and expressed in percentage Seed yield Harvest index (%) = x 100 Biomass

3.11 Seed analysis The seeds collected at harvest were chemically analysed for the protein contents. The seed samples of each treatment were dried and ground to fine powder and passed through a 72mm mesh sieve. The powder was stored in the polythene bag with proper identification. Before analysis, these samples were kept at 80°C in an oven overnight. Estimation of total proteins The method of the Lowry et al. (1951) was followed. 50mg oven dried seed powder was transferred in glass centrifuged tube, to which 5 ml of 5% trichloroacetic acid was added. The solution was allowed to stand for 30 minutes at room temperature with thorough shaking for the complete precipitation of the proteins. The material was centrifiiged at 4,000 rpm for 10 minutes and the supernatant was discarded. 5ml of IN sodium hydroxide was added to the residue and mixed well. It was left for 30 minutes on water bath at 80°C so that all the precipitated proteins may completely get dissolved. After cooling for 15 minutes, the mixture was centrifuged at 4,000 rpm for 15 minutes and the supernatant containing protein fraction together with three washing with IN NaOH was collected in 25ml volumetric flask. Volume was made upto the mark with IN NaOH and used for the estimation of proteins. 1 ml

56 Materials and Methods sodium hydroxide extract was transferred to 10ml test tube and 5ml reagent-B (Appendix, p.iii) was added. The solution was mixed well and allowed to stand for 10 minutes at room temperature. 0.5ml Folin phenol reagent (Appendix, p.ii) was added rapidly with immediate mixing. The blue colour developed and was left for 30 minutes for maximum colour development. Absorbance of this solution was read at 660nm. A blank containing DDW, reagent-B and Folin phenol reagent was simultaneously run with each sample. The protein contents were calculated by comparing the optical density of each sample with calibration curve plotted by taking known graded dilutions of standard solution of Bovine serum albumin (Fraction-V) and the seed protein contents were expressed in terms of percentage on dry weight basis. Standard curve for total proteins 50mg bovine serum albumin (Fraction-V) was dissolved in 50ml DDW, of which 10ml solution was diluted to 50ml. 1ml of this solution contains 200^g protein. From this 0.2, 0.4, 0.6, 0.8 and 1.0ml solution was transferred to 15 test tubes separately. The solution in each test tube was diluted to Iml with DDW. A blank of 1 ml DDW was also run with each set of determination. 5ml reagent-D to each tube including blank was mixed well and allowed to stand for 10 minutes. To this solution 0.5ml Folin phenol reagent was added and mixed well and incubated at room temperature in the dark for 30 minutes. Blue colour developed and was read at 660nm.

57 ^sults CONTENTS RESULTS

Page no. 4.1 Experiment I 58 4.1.1 Growth parameters 5 8 4.1.2 Physiological parameters 62 4.1.3 Yield and quality parameters 65

4.2 Experiments I and II (Pooled) 66 4.2.1 Growth parameters 67 4.2.2 Physiological parameters 72 4.2.3 Yield and quality parameters 74

4.3 Experiment III 76 4.3.1 Growth parameters 77 4.3.2 Physiological parameters 80 4.3.3 Yield and quality parameters 82

4.4 Experiments III and IV (Pooled) 84 4.4.1 Growth parameters 84 4.4.2 Physiological parameters 89 4.4.3 Yield and quality parameters 92

4.5 Experiment V 94 4.5.1 Growth parameters 94 4.5.2 Physiological parameters 99 4.5.3 Yield and quality parameters 101 Chapter-4 RESULTS

4.1 Experiment I In this factorial randomised pot experiment conducted on chickpea (Cicer ahetinum L.) cv. BG-256, the comparative effect of two irrigation waters and four levels of fly ash was studied. It is to be noted that Experiments I and II are also expressed through the histograms as the data of these two experiments was presented along with the pooled data. Only the significant data are briefly described below (Tables 6-25; Figs. 3-9). 4.1.1 Growth parameters Twelve growth parameters namely shoot length, shoot fresh and dry weight, leaf number, leaf area, branch number, root length, root fresh and dry weight, nodule number, nodule fresh and dry weight were recorded at vegetative, flowering and fruiting stages (Tables 6-17; Figs. 3-6). 1. Shoot length plant* TPPW increased shoot length and it proved superior over GW at all sampling stages. TPPW recorded an increase of 16.60%, 17.24% and 17.62% over GW (Table 6; Fig. 3a). FAio concentration of fly ash proved best, which was followed by FA20 and FA40. It recorded an increase of 15.39%, 16.07% and 15.59% over control. Regarding interaction effect, TPPWxFAio proved best as compared to other interactions at successive stages of sampling. TPPW^FAio showed an increase of 33.86%, 35.64% and 35.00% at vegetative, flowering and fruiting stages respectively when compared with control. Combinations of TPPW with various fly ash concentrations proved better in increasing the shoot length than the combinations of GW and fly ash. Among GW combinations GWxFAio gave better results. Shoot length increased consistently from vegetative to fruiting stage. 2. Shoot fresh weight plant"* TPPW proved superior in enhancing the shoot fresh weight at all grov^h stages. It registered an increase of 19.86%, 20.70% and 20.19% over GW (Table 7; Fig. 3b). Among the various concentrations of fly ash FAio proved best, followed by oo »ri ON CS o lO O ON ON Mi 00 ro •^ •rf o ^ cN O Tj- Tf r~ ON ON CO (S 00 »ri CO c o o o o o o — en m o o o o o o o ca. •4—» o r- wo vo ON ^ O IS 00 CO NO IT) .£ ^ •^ '-' CO •— >/~i •—I o m r~ o CO >—' 'C •^ •^ lO r^ ON ON CO ^' vd •^' o o o o o o -^ O m m fo Q o o o o o o o a •*->o 00 NO t^ u a. C fS 0\ O > se >ri ON t-~ CN NO f- t- »-i O 00 -o Uc r-

, X X X a- O ^-^ CO > ^

C I s 3 Tt CO r- ON ^ 00 NO t^ > DO (S

O ea 00 00 CO ON 00 '^ ^ t-~ ON 00 00 CO VO CO (S CO •<;r >n O O ON ON ON NO CO co' r-' W-) ^ NO •<;r o r- ON 00 TJ-' CO CO CO (N (N CS •^ CO Tl" WO WO x: «-• Si >^ •^ ON t^ (S ON — ^ ^ NO r- NO (N ON '^. "^. ^. \oON 00 ON ^ wo •«}• wo 5 « Tt 00 NO VJ CN CS (N fS (N oo' w-i q ^ q r-' —: vd o 00 CO CO CO H ^ c NO" CS ON wo vo wo 6 CO Tj- CO c '5 n o o 0) CO NO >ri >n (N ^ ON o (S CO 00 CO Tt CO wo vo vo a. c o Ui c «* o 00 m CO aoj ON ON trt NO -— o vo OO O Tj- WO CO O CO Tj- ON ^ ON 00 t~- r- 00 •rr ^ co" f-' iri 0\' b (N « CO TT CO

•4-* ac> 3 O c c c3 tEi V ?? CO ID 6^S 5 H 80 CD. at 5% Vegetative Flowering Fruiting Water 0.080 0.063 0.072 70 Fly ash 0.113 0.0«9 0.102 Interaction 0.160 0.126 0.144

mii:z*

CD. at i% Vegetative Flowering Fruiting Water 0.061 0.067 0.066 Fly ash 0.086 0.094 0.094 JS 0.133 "a. Interaction 0.122 0.134 00

I o o .s tn

S 8 •5. to t

Vegetative Flowering Fruiting (Experiment I) Fig. 3. Showing the effect of wastewater and fly ash on BG-256. The data is presented in Tables 6,7«&;8. t4- ^. c —I O NO 00 as t~ ro NO _ CO o m m m m r«i 00 M O -^ t-- f^ NO •^ '-" C IS n m m o NO 00 E o tu r-^ 00 i> •-^ m (S •f-* O •^ ^ NO 00 oo — I—1 1—1 1—1 o u 2 U-i '"' CO o O '— '« o o o o t/5 o Q o o o o o o o o CO O b o a. — M .» r^ ^^ •^ -o ° f-^ > _e J3 Tt O CN NO ro O c S <^ "O NO '••-» Tf m NO — m CN CO V) CO ji h O r-' o\ oo" (S >o rr *"• ^ Tf TT NO c^ 00 C/3 C/5 o o o o o B O OO o o > 6 ii o o o o o tu ^ ca > a. V o o ^ VO (N rr C~- ON 00 NO fo en ri r^ w~> ONN O < O- J3 w-i oo' NO V3 ON —<' O o ^^ ^-H 1-^ l-M i-H 1-^ c n Tt o X c ON m NO o —' r^'

j= C M u —« U V CO J5 > o ... • •M u TT •rr ON O r- ON •^ M o CN O r- rr NO vn r'l _> r>- 00 CN o _C NO O 00 fo iri T^- NO <^ '^. '". ^. NO NO C4-1 •a 'Z •4-* > •= '5 > < < 0 0 E " O CO CO r- 00 l/-^ •"^ -^ -^ ON o >n CN -^ rr CN 00 __ < rn .—; t~-- ON CO NO '-' < m in o NO r~ — NO Tj-' w-i rf NO' 00 r-" NO ^' r-' NO •^' ON r-' ^ ^ Pi ^ ^ 1 ^^ — t—1 CM CN CN CN « "a. T3 P 0 g- § & V en O ^ 13 CO '^ NO m ro (N f^ ON 1— ^ NO CJN ^^ in NO 60>-J (N 00 »ri 1—1 .—1 NO in -^ « 9. ^ -k-* "7 i2i -SJ Q. CO •C -n w s c ^ r-' u 4# ^^ s O^S b^S 2 l-l b^S ^11 2 H s Results

FA20 at all sampling stages. FAo was statistically at par with FA40 at three stages. FAio registered an increase of 35.70%, 36.60% and 36.93% over FAo. Among the various combinations, TPPWxpAio proved best and gave an increase of 62.57%, 64.66% and 64.29% over control. Among GW combinations, GWxFAio recorded maximum value being at par with TPPWXFA20. It indicates the utility of FAio whether it is applied with TPPW or GW. TPPW-FA combinations gave better results as compared to GW-FA. Shoot fresh weight showed an increasing trend from vegetative to fruiting stage. 3. Shoot dry weight plant* TPPW proved superior by giving significantly higher values at the three growth stages studied (Table 8; Fig. 3c) and registered an increase of 15.87%, 15.28% and 16.73%o over GW at vegetative, flowering and fruiting stages respectively. FAo was found to be statistically at par with FA40 at vegetative and flowering stages. FAio recorded an increase of 29.89%, 32.05% and 31.33%» over control at the three sampling stages respectively. Among various fly ash and TPPW combinations, TPPWxFAio proved best at flowering and fruiting stages and recorded an increase of 50.36% and 51.84% over GWxpAo. Minimum value was recorded by GWxpAo which was at par with GWxFAto. Dry weight increased consistently from vegetative to fruiting stage and it may be noted that increase was comparatively more between flowering to fruiting stages than vegetative to flowering stage. Fly ash concentration of FAio proved beneficial when interacted with TPPW as well as GW. 4. Leaf number plant"* Irrigation by TPPW proved effective as it increased leaf production by 40.93%, 41.60% and 39.19% over GW at three respective stages of sampling (Table 9; Fig. 4a). Fly ash concentration of FAio also enhanced leaf number in comparison to other concentrations at all sampling stages, whereas FAo gave the value which was at par with FA40 at flowering as well as fruiting stages. Among various combinations, TPPWxFAio was found to be superior over GWxpAo at all sampling stages. It registered an increase of 93.23%, 96.66% and 93.02%. The lowest values were recorded by GWxpAo and GWxFAjo as both were found to be statistically equal. It may be noted that TPPW-fly ash combinations proved better than GW-fly ash

59 00 00 c 60^ o 00 m lO CS -"r "^ •^ VO 00 u c —' /^ m r^ ••-» fS o o o o o o o k. o o o o o o o c c u-> m 0\ Ti- NO ^ o r- ,^ O (S m ro •^ «ri TT t3 a> ^ o '—I m m ^ c/D i 60^ 0 0 0 0 0^^2; > o o o o d O < n Q. VI ^ VO "^ O < c^ 0\ ^ CS 00 a. m' m' m »n vo w-> (L> 60 o o o 00 00 f) r-- ON ro u c l/^ O 00 J-, vo -^ m X o (1H ri M (N Tf •^' •*' en « fe 03 g O O OQ o •g ^ X >, X X X o a- > ^ <: E < m < a, 1/1 b s XI « r-

C - S 3 un fo VI O 00 O Ov 0\ c 00 — t~» NO > M c4 r-' 00

X 60

o o o ro fO 00 O t~~ ON (S TT r«^ r- o 00 Tj- ^ r-- ON ON rj- — m CN T}-' in •q- NO NO < gj CO vo 00 r- CN ON VO oq NO (N ON •T3 "" > ON — O o o >n iri —. n <^ so c NO r^ 00 O — '- — IT) —' CNJ CNJ NO •<5- w^ ? Tf in '<*•• NO t-~-" 3 -C K rt « > w •*—• T* CL « •C X W 5 S;^ 60 X -^ 00 c 3 O £ 00 —1 vo CN ON o rt ON m 6C J w (N -^ .—1 m ^ r~ 00 00 NO ON CN in in ^ s n <^. 2 .—1 1—1 1 o o .—• (U b ••-»

o o\ o w-> IT) ON (S (^ 60 c^ r~ ^ ON Tt t^ -^ «-i C >^ ^ O CN ON »n t~ 00 u b rW vd Tf ^ NO •^' ON 4-* Tr o -^ o m c?\ cyo s3 o >i-) t-~ VO NO 00 r- NO r- r-~ 00 •^ ^. ^ Z 'So Q o CO o o o •4-i «3 O b O &c o • PH t- vo r- ^ (S NO O — m NO NO CO U-) '^ Pu o r-' /^ 00 \0 VO ON 00 r- u O CN CN CN 5 o o o o ZZ"^ o o > 60 < a. > o VO O ro Tt 1—1 m en o r-; < T3- 00 ^ ^ r~. o\ a. "EH H r*^ (cU" X> ^^ 60 B o 3 ^>-^ /"^•^ 4-» c CO VO O 00 Tj- NO 00 O O 'S C4-I cd ,— T3- fO m r~- o X J) fo «r> Tf" ON o 1^4 r~ \o ^ vd •^' CO U U OQ c NO 00 t^ NO v> o XXX o > ^ < b < m < •o "cO ? t/5 OH CO E u<

-4-* rt ^ 5 u c as — m ON c m m r- -"^ 3 "5 r^ CTJ fS tS 00 m CO NO m vo NO < (U ^ r~' m' Tj-' CN NO NO' O ^ 60 Tj- in TJ- NO r~ o 00 CN 'cO *"• J3 1=C : -c/2i *—' JS j= 60

o o ON (U O ^ Tt 00 m (N 00 m ^ r- NO NO ^ m c 00 ^ ON ^ 00 Tt CN — o o TJ- t^ NO 00 •—N ^ 60 r-' csi •*' ^ r-" ON K b NO' NO' .—i' r-' K CN '^ < as CO in '^f Tt «n •^ "cr NO C3N 00 r- o C3N 00 b ^ t« CO hs :«<: 1* _ v o • PH m 00 ^ CS ON NO m 60 o • MM >n CN • NO r>- r^ o JS NO '•4J b o «n r-" •^' CO ^' O b •a O rf r~-' in r--" ^ O 60 G >» ••-» u n 'a- NO —c CS '"' 60 > 6 b > 6 > < PQ < CQ o o E "^ o (S TJ- f<1 O 00 ON *—1 o — -^ oo -^ 00 NO r- CO CO ^ t^ ON • « •J C g ^ O- CO ^ 2 1 •c j= W 60 •4-» b^ 60 c o\ cV

C.D. at 5% Vegetative Flowering Fruiting 1400- Water 0.080 0.1 S2 0.126 Fly ash 0.113 0.216 0.178 Interaction 0.160 0.305 0.250

Vegetative Flowering Fruiting (Experiment I) Fig 4. Showing the effect of wastewater and fly ash on BG-256. The data is presented in Tables 9,10«&11. Results combinations. Leaf number showed a linear increase from vegetative to fruiting stage. 5. Leaf area plant' Application of TPPW proved superior over GW as noted in other parameters (Table 10; Fig. 4b). TPPW secured an increase of 23.88%, 23.20% and 24.39% over control at vegetative, flowering and fruiting stages respectively. FAjo fly ash concentration enhanced the leaf area, whereas FAo gave the lowest values at all the sampling stages. Regarding interaction effect. TPPW^FAio proved best and closely followed by GWxFAjo indicating the utility of this dose whether applied with TPPW or GW. Every combination of TPPW and GW with fly ash significantly increased the leaf area over control but TPPW-fly ash combinations gave better results than GW-fly ash combinations. Leaf area increased from vegetative to fruiting stage. 6. Branch number plant' Like other parameters, TPPW proved effective and showed an increase of 20.60%, 24.97% and 25.86% over GW at all sampling stages respectively (Table 11; Fig. 4c). FAio fly ash concentration proved best in enhancing the branch number at all the three growth stages, followed by FA20. It recorded an increase of 53.98%, 51.41% and 54.36% over control at respective stages of sampling. Among the various combinations, TPPWxFAio proved best at three growth stages, followed by GWxFAio which was found to be at par with TPPWxpAjo at flowering stage. It may be noted that GW with FAio can also be applied to gain better results regarding this parameter. TPPW^FAio recorded an increase of 87.25%, 87.39% and 93.70% over GWxpAo. Branch number also showed a linear increase, however the increase was more pronounced between vegetative and flowering stages in comparison to flowering to fruiting stage. 7. Root length plant' At all the sampling stages, TPPW proved superior over GW giving an increase of 21.63%, 21.03% and 22.15% over GW (Table 12; Fig. 5a). FA,o concentration proved more effective and it was followed by FA20. An increase of 35.52%, 36.67% and 36.04% was registered by FAio over FAo at vegetative, flowering and fruiting stages respectively. Among various combinations, TPPW^FAio proved best at all sampling stages followed by TPPWXFA20 at vegetative and fruiting stages and

60 o cs C3V r-~ o in •^ ^^ m m •^ O o o t~-' o m oo C3V m vo r~- •'^ •*-> 00 -—I 00 t^ o o o •-< —I •—I cs IT) r~ t^ Ov •5 o o o o o o o

>-» 4-> a r^ 00 — 60^ CS m 00 o ov o C f) 00 —' .—I 00 00 00 ^- m vo o o us •4.J 'C ^^ CJv Ov •^ 00 OS vo •MM O cs r-" o *rt n) o o o — —' — cs u-i vo >ri t~- cs u-i vo o o o o o o o o «3 vo 00 r-- Q a O E O (L> a. o, CS 00 >n m m Ov CS j^ 00^ > c o C ^. ^. ^. VO vo Ov cs r- IS •* en ON V3 vo w-i vo' r-' cs r~ 00 c^ ^r m vo o «o oo ^- CS vo •^ m (T) vo vo t~- c^ 00 m ^ vo vo o m •^ Ov o r~ O O O t—< f—I »—< I—I 6 o o" o o o o o

»n 00 vo •—' C3V m o m 00 o O vo «/n O o ^ ^ ,—1 . rr •^ 00 vo o c •n CONv 00 o cs ^ o (U CO o I cs u-> vo — •<;r •PQ vo O 00 en «r> Ov C3 «Ai K ^ fern g u u m O ^- m f^ CS Tf I >n vo »/^ ll Irt X ^ if V if VO 00 r- X X o g P3 <

Cu « cm X) r- C t- a 00 Ov IT) O 'c O fn ^ vo o t^ Ov r-« c^ TT 3 OV O K vd r-' cs o —' CS 00 00 in 00 •^ m 00 •^ *-> cs cs CS m vo oo ov — SO

ro cs oo O O ITi m cs m 00 O -^ 4_> M cs •'J; TT m m r~- cs Tt oo' Ov 0m0 m CO 00 cs vd ^' ov r-' 00 Ov cs vo 00 in vo < O -"S- fS TT o r- o o o cs r<) cs cs m cs m r^ vo 00 O CJv w « o « «n cs cjv in vo •—1 ov cs ^ o O m > vo r^ .— 00 r- 00 vo C3V O 00 rr" —. vo Ov m r-~ o <*- 00 Tf —I o •-' in cs >/^ •^ CS CS CS CS m m 2 o '-' f^ ri ov — O vo 00 t-- 6 O 60 *^ CO = 1 ^-^ > in r- — vo cs C3V o m 00 Ov TJ- Tf vo m O —I Ov VO 00 ov ^ vo m m E ° *i •<-• cs" 00 in CS «n Ov vo' O 00 cs cs cs cjv »n cs r^ vo —I c^ o ov vo vo ^

•o k1 Ov Ti- r~- o o cs ov vo vo TT m VI 60 cs 00 irt r- o "*. P '~~. (U (4-1 <-• >ri rW 00 m r~ vd oo' cs in cN m m — O o •^ O -^ CS •«r o r- Ov CS vo O o o oo t? •«-» c CS cs cs CN m cs m r~ vo 00 O Ov r- > o ^ CL 'rt ^,1> o W 5b

3 O OO cs X3 m o o\ ON

cd o vo .— Tf 60^ _C r- t^ r- 'u, ON ON ^ •^ vo ON vo o •<1- »r) »r> 4-J ^ C C ^ r- ON vo vo ON OS —• O ^^ tS n •^ m m 00 *c 4-* o o o o o o o o o o o* o o o o O g OQ o •* -^ -^ ON •—I O r- ^ •* m •^ ON o vo 00 r~-' ^ Tt (S c 60 O c o ON in cs — o NO ro «o •aa Tt «n «o K ON 00 tJ fe CO 53 o o cQ •g ^ X >, X X X o > ^ < E < CQ < CO l-l U "3 V bO cd cd X) ts 4-* ^ n ja 4-u* CO (4 ^ c2 ^ 'c 6s0 •4-> 3 0) ro 00 TT ON ON (N OcJ 0) NO ro fO ON cs cs m •^ vd r-' cd td x: ^ 60 o ^ o. 0) vo lO NO ON 1/^ (N vo '^ «n (S vo ON ;>^ •^ r- lO "3 CS -^ m (N o r- •* r- r>- tN XT, vo' vo ON —• O 00* < E o (S tS (S r^ CO fo a> (» 4-^ "«> >• to •o 6i cd c VO ON 00 ON vo fO r-- r^ f- >^ > vo fO O Ua^i fo" rf' T}-' vo t^ c-~ vj i (S IT) ^ T^' (S O 60 B •^ E % 60^ Vw^ •o = '5 J3 n n E Z r~ Tt vo —I fO (N o m r~ cd Id '". ^ '^. r^ t->- ts ON •^ —: fN ^ C « -O f<-i rn CO Tt" «ri >ri 00 ON 00 Tt 00 vo' ^ 4-* .Z^ c 2 « "o- 3 60 O c '^ i-i 60J •a td ro ts r«i vo r- ON <4-l <—I ON O vo (S 5 ^ rn ; vq - o (S « JS 4-» *7 W Q a. cd ^ 60 c 3 O E C/3 (N OQ u 2 o \o 60^ o ^ -"^ m "C ^ ^ —' •—I CO K o '•4-» ^ vo NO 2^ -^ c^j r- •*-» ^ CM O o o o 03 , <" (U a, O m m fo VO t^ (N 60| m ^ CN CO Tt ON c 'C (N NO ON CO O in NO O (NJ ^ in 00 vd u 00 r4 »r> ON ts ^-< NO c« o o o o —< ^ — CO CO Q o o o o o o o CO •*-> U o o ex, o vo •^ ON r~ ON OO ^ —• NO c — ex, '•^ CO W-i Tt O »r> c^i •4-* c" cs (S r^ ID o 60 ko> O c O ON O 00 ir> t-~ U o CO O CN r-H CO r>; CQ x: X O CS ^ iri od NO CO CQ 0) ? CO U O CQ CO CL, IICO X >% X X X OH O H^ < U. < CQ < CO U CO 60 +-» cd CO J3 1 \<2 rt O C ^ 00 3 4-* O CO NO NO u o o OO CO c I-uI o o J2 ON —' — CO "S. 00 ^ r-; CO IT) ^^ cs NO CO O ON 00 ON CO NO ON c4 <4-c 00 O ON (N «r) ^' o csi —• NO ON 00 I <

Ui o C/}

CO •4-* > •o JU CO O »/^ CO CO o r-- ON >-^ c •4-* CO t^ ^2 o — CO > -^r o fNi ^ c ON ON ^ Ml rj- Tt CD (N I CM 1^ ON — O V) O Uui 00 NO 00 NO CO CO —; CN r> 00 00 0) «>w 00 O ON NO ON t-~-' C cs >o CO o fs ^ 1.2 -o *-• ••••1. u > « ^ ex d W e o 60 (N 1 CO 3 O 00 (N c g| CO CQ 30 n CD. at 5% Vegetative Flowering Fruiting Water 0.113 0.119 0.105 Fly ash 0.160 0.169 0.148 25 Interaction 0.227 0.238 0.209

CD. at 5% Vegetative Flowering Fruiting Water 0.0068 0.0176 0.0054 Fly ash 0.0096 0.0249 0.0077 r Interaction 0.0135 NS 0.0109 .60

0.6 CD. at 5% Vegetative Flowering Fruiting Water 0.032 0.074 0.052 Fly ash 0.046 O.IOS 0.073 i Interaction NS 0.148 0.104 .60

O^^

GWxFAio at flowering stage. Lowest value was recorded by GWxFAo and GWXFA40 at all growth stages, as both were equal in effect. The increase was 21.63%, 21.03% and 22.15% by TPPW^FAio over GWxFAo at respective stages of sampling. Fly ash concentration, FAio proved superior in comparison to other concentrations as it gave better results with both the irrigation waters. Root length increased consistently from vegetative to fruiting stage. 8. Root fresh weight plant' TPPW performed better than GW in enhancing the root fresh weight, giving 18.35%, 19.13% and 18.83% more values over control at the three stages of sampling (Table 13; Fig. 5b). Like other parameters, FAjo concentration of fly ash proved best at all three stages in comparison to other fly ash concentrations and gave an increase of 23.76%, 24.75% and 24.98%. TPPWxFAio proved best at vegetative and flowering stage. An increase of 46.73% and 48.70% was recorded by TPPWxFAio over control at vegetative and flowering stage. FAio fly ash concentration also proved beneficial for this parameter when interacted with TPPW as well as GW. Root fresh weight increased from vegetative to fruiting stage. 9. Root dry weight plant"' Application of TPPW proved superior over GW at all the three growth stages and showed an increase of 53.28%, 52.94% and 52.68% over control (Table 14; Fig. 5c). Fly ash concentration, FAio proved best in enhancing the dry matter accumulation. Significantly this parameter also decreased with the increase in fly ash concentration throughout the sampling. However, FA40 was at par with FAo. TPPWxFAio proved best while lowest value was given by GWxFAo which was at par with GWXFA40 at all samplings stages. TPPWxFAio obtained the increase of 130.77%, 131.03% and 131.60% over control. All the TPPW-fly ash combinations gave better results than GW-fly ash combinations, where GWxFAio was found to be at par with TPPWxFA« at vegetative and flowering stages, even TPPW alone gave better results than other GW combinations. Root dry weight showed an increase from vegetative to fruiting stage. 10. Nodule number plant' TPPW performed better than GW for this parameter also. It registered an

61 i^ c m m NO m to 0 CO OS o o NO c ON CS m (N VO 00 . 0. z CO 1 0 0 u b 0 d d d r^ 0) n. CO o W-) ,_ 00 f«^ t^ o •«r 60 O C •0 _C J3 ON 00 00 C ve o en ^ Tt r~- 00 ON CO c ^ ON VO m '5 •^ JS fe o o V) cO 0 0 0 0 0 0 c/0 c/i "a. u O •—' '"' '—' '"' '"' •"" 0) 0 0 0 0 0 0 ;z; 00 0 0 5 •4-* o > 6 d d d d 0 Pu < > JZ 60 n a. C/0 5 'S o O ON U~l ON ON •^ 'cj- yr) vn m ^-( CS r- »-^ JS < m o 0^ 0 'c u en o o •^ CN VO ^ o •* CS CS X ON f-^ in CS JS 0 C b o n '^ C/3 o ^ CQ CO 0 0 OQ -, XXX 0 •^1 T3 > ^ < b < CQ < OH CO "eo H -^ 00 •^ ON 2 >^ 00 Ov ON CD fS »n r~- VO ^.-^ o b m < d o o -• -• b e O o o x: 0) V t/3 JC > CO •^ JU V o .„ o ON vrv CS NO ON t- 60 = •«« ,^ ^^ _ Tf m ON VTi JS VO 00 CS CO CS CS _C J3 r—( f-^ 0 — m 00 _> r~ < 1—t ^^ VO c^ "S -a '•3 -^ "O 00 oo so o CS OS '•4^ "O r^ CS VO Os t-- •^ Sra Q^> CO r- Ul 0 V) 0 60 k> b o l-H f—* b 1"H 1—H »-M r^i *ri u o o o ts «"^ o •5 U •—' »-^ CS »-^ j_. C 5^JS 60 > 6 b > 6 > < n <: oa 0 0 •- .'ti o f—C n •^ 00 CS 0 t~- J2 OJ ^1 o o I; b —" CS CS -o p c « 3 60 a> en O ^v •0 rt 60>-) m fS ro en o CS r~- U-) 0 00 m C3N t~- cs c« ^ n m •^ rr 3 -C o 5 b t- oo c^ •—; o ON b 0 CS TT r-- VO m — 0 o o o O ^ ^^ o ^ '-' ^ ^ -^ ^" ^ 9. ^ > « 52J > 0. CO

M 00 r<-i ••-* c *—^ i>a u 3 0 a> B c c c E c ^ c c 00 rv) CO CO CO CO 13 ^ ^ ^ ^ to ^ ^ CO •C2O ^^ s od O ^s O ^S s 0 ^ s b^S S 2 00 •^ 00^ o o Tj- ^ r^ NO CS r-1 NO m NO ^^ •—I >—I (S ro CO •^ (U o o O O O O O O O G o o o o o o o o o d d d d d d d

•4-» o VO VO ,-1 IT) fO ON •^ ON r- O >—' >/-) 00 '^ IT) 00 •^ NO ON o 4 'C O r— r-H CS CO CO -^ PL, o o o cs m f<^ VO O CS NO NO o c^ 00 C < VO 5N ON ^ c4 c cs m fo CS O ^ CO CO CO rn c -I fi -: 1^ o o" o o o o o o o EG 00^ o o o 6n o o o o o o Z d d d d d d '5 o CO cs ^ cs u-> ON t^ O •-' ON «0 O NO 00 CS fS (N CO •^ ro O. o o o o" o o c

•*-» DO o O o c r> b o" o o cs ri cs eg u u m CO o o o J3 ^ X >, X X X O CL, •o OH > ^ < E < OQ < (_| CO CO 60 a I f Xi

to O c c .—1 CO (N r-~ o r- NO CO 3 ON (N CS 00 CS O CO '— = b o — >—1 t—i CS CO CO >o o o o o d d d d

CO 60

^ t^ Tt r-» m w-> ^^ >n oo ON CS NO 00 O ON O NO fO c^ NO CS NO ON o ^ o '- CS CS CO CS < gc3 «j^ o o o d d d d d d o o o CO c<3 (U O CN NO CS r~ «r> ON NO CO NO CS ON >> > J3 ON CS O CS 00 »n ^ O NO CO O 5= «J § o ^ ^ CS CO CN CO m (4-1 O o d o' d d d d d d d d d O 60 bO o ^ < CQ CQ o o £: •-' NO NO NO NO O 00 CS U-) ON ^ r~ ON CO ed >n •/^ fo -^ CS — CS CO CO TJ- NO in *l d d d o o o d d d a. •o o d d d u a. c ^ O cd o ^-^ c V CO bOnJ 00 /^ fO 00 NO CS oo NO CO > o I b o —' o *—< CS CS CS CO d d d d d d CO J3 d d d d d > «>

O. CO u CO 60 X5 c 3 O 6 C/3 CS ^1 CQ Results increase of 28.00%, 11.16% and 29.15% over GW at vegetative, flowering and fruiting stages respectively (Table 15; Fig. 6a). Among various concentrations of fly ash, FAio proved best at all the three growth stages followed by FA20. Nodules were also affected adversely with the increase in fly ash concentration thereby FA40 giving the lowest value which was at par with FAo Regarding interaction effect TPPWxFAio proved best in comparison to other combinations followed by TPPWXFA20 and GWxFAio at the three stages. Nodule number increased from vegetative to flowering stage, but decreased at fruiting stage. 11. Nodule fresh weight plant'' Nodule fresh weight was increased when treated with TPPW at all sampling stages giving 18.30%, 19.23% and 21.61% more values over GW (Table 16; Fig. 6b). FAio proved best as it increased the nodule fresh weight by 30.48%, 31.12% and 33.21% over control. However, at vegetative stage FA20 and FA40 were at par on one hand and FA40 and FAo on the other. Among various combinations of TPPW-fly ash, TPPWxFAio proved best. Nodule fresh weight showed a gradual increase from vegetative to flowering stage, but decreased at the fruiting stage. 12. Nodule dry weight plant* Between the two waters, waste water proved efficacious at all the three growth stages giving an increase of 25.35%, 25.38% and 25.00% over GW (Table 17; Fig. 6c). FAio proved best in comparison to other fly ash concentrations at all sampling stages. Nodule dry weight decreased with the increase in fly ash concentrations after FAio thereby FAo gave the lowest value which was at par with FA40. Among various interactions, TPPWxFAio proved best at flowering stage only as the data of vegetative and fruiting stage was non-significant, whereas lowest value was given by GWxFAo which was found to be statistically at par with GWxpAjo. It may be pointed out that nodule dry weight increased upto flowering and showed decline at the fruiting stage. 4.1.2 Physiological parameters Physiological determinations namely leaf nitrate reductase activity, total chlorophyll content and NPK were investigated at vegetative, flowering and fruiting stages (Tables 18-22; Figs. 7a&b, 8a,b&c). The data found to be significant are described briefly as follows:

62 o m t^ 00 00^ o \0 '—I c 00 VO VO m r~- —' oi 00" Ch vo iri o m m NO r- 00 •—I 000000^ CT3 vo 00 U( o" o o o o o" o

••-»

03 o r- u-i ^- 04 UO TJ- 00 IS r-H TJ- 00 ^ ^ \0 ON CS '^ 00 VO '— ON o to" •^' ON ^- ^ ^ r^ ON O CO ro '^ ro 00000-^-^ o Q o o" o o o o o -4-» U o c •a c cs vo 00 r-- > c : NO O O kD ON ^ ^ ^ O •^ ^ CO 00 00 fo «fi r- rr «o O Tf TT 00 O O •^ fO •^ •^ (S vo 00 t^ 0000—1—'^ o 6 a«);^ o" o" o" o o" o" o S > in O o •^ Tf ON «n ON (S ON tS O •o Tf «n «n t^ O ON o c 00 o e o ^H 00 »r) 00 ON ON c O O «0 ^ *^. f^. o «0 •^ ON m •^ fO ^ b OQ "I o u m , X X X O > ^ < UH < CQ < TD

X> E u ^ a 'c ^ NO r- o CO »0 t- o •«*• o o

"0.43 tS 00 O VO •^" •>^' cd 0) CO 00 ex '^ ^ o ^ 00 r- f'l >n 00 r- m ON NO r- ^ -^ f- TT '-< — ^ VO ^^ ON m VO O NO rW rW r-i 00' ON TT" ^ •—«" ON" «O 00" CS (N (N m •^ f

(/I E o > 00 ON ON VO — Tf •^ 00 ^ •* ON CN o. «=>. 9 so f^ 1— r^ m o cs r>. VO r-; «*- CO CN| 00 »0 Ml vd r-' ^ m m 00 o 00 •*-» CS «N Tf c c «3 •si U Vi W-) ON o 00 « a. cd

3 O J) CO z CD. at 5% Vegetative Flowering Fruiting Water 0.076 0.071 0.073 Fly ash 0.108 0.100 ^^0.103 Interaction 0.153 0.141 110.146

0.00 '^ O^^ •^ v'O Vegetative Flowering Fruiting (Experiment I) Fig. 6. Showing the effect of wastewater and fly ash on BG-256. The data is presented in Tables 15,16&17. O (S 00 'a- 00 ON ^^ vD O m c Tj- m r- r~ (U en O O O O K-0_0 ••-» O O d d o o o o Z nS Ui o o o o o o x(U: ex -•-» -•-» ON -^ r- Tf I/O O oq ON w-> (S O ON liO m _c CO m in VO ON ro e u CO to ro m m NO m >—< ro ro m r~ r~~ c < d d d d d d o o o o o o c/D 00^ o o o o p p ^ J5 d d d d d d o • SP o pa > CL, 'S t/i fS W-) ON 00 VO r~ o. CO c' d d d d d d u 00 o 3 o 00 (S UO M- (S fO WOQ U U ON wo CS ON ON TT X C! (S CO CO Tj- in m J3 C O d d d d d d m s u u cQ (/I X X o > ^

CO

•4-»

c2 ^^ u o c «5 00 VO cs CO O ro cs 3 ON (S O VO 00 cs r~ ro — cs CO ro — cs cs ro d d J :^ d d d d d d ccj 00

r~ ON ro ro •^ ON ON cs VO —' ro r- t^ O ON 00 CO O VO O 00 m o r- CX"+3 — ts — (S ro ro —' cs — rs ro cs _ t*- d d d « O d d d d d d d d d

(U o a >. ON m t^ cs o ^ oq ON 00 ON VO ro m O VO ro VO cs ON oo M o ^ fO CO fO c !:: ;a 2 V© CS ro CS <*- _ (M />o ^ d d d o o o cs d d d o 00 . ^. ^ VJ s *- .S O O O 6fS § ^ b CQ ir o CN CO ro O 00 rr t^ C> 00 00 cs m cd CO cs VO rr VO (N ON — in ro cs O VO 4} T3 cs c4 cs ro TT ro cs cs cs CO ^ ro £ U 4-» ••- d d d d d d d d d d d d OJ O. •a p 3 00 o /-^ Is 00 1-1 «n VO <—1 r^ r^ CNj VO Ov ro VO —« Tf c« > C~ O ON r- r< o •^ o r- « CS —I cs ro ro VO <7N 00 CN CO fS Si ^ d d d d d d o o o d d d - "eo JZ o 00 x> 3 O 00 cs

to O 00 -—I o m o —' CO o o o o ^ 2 Z 2 2 Z 2 l_ C3

•*-• ••-» CO o •* C- vo •^ fn a\ bDi, r-- o\ 00 ^ •^ {S c IS O in rr> VO Ov o 000 'C 00 c/D o o o o o o ^ O E5 O O C/3 O O Z O O 2 Z t« o d d d d CO •*-» bO o O cx ^ VO Tf 00 ^ U-) > •a p 2 2 vo CN VO Tt "tj- >0 O CS 00 00 c R3 r- O — tN

rj- 00 ^ rt 00 \0 o O C<1 CN VO O 00 ^ cs ^ •O O O O c o o o 60 3 O T3 O m vo >/^ 0 0 C r» ON 00 ts ^ r^ c 000 '^ Tf CS o o o o o o o S cj cj m H) :^ X X X o > ^ < tju < m < -o "eO en CO

to X c2 ii p C 3 «! 60 VO O -^ OS 00 00 o »n t~» 00 fo •<;»• VO r~ O O 4>' l-i o o o o o o C JS o o o o d d d d CO Rt *' CXi CO 60

>-< x; ^- f<^ r~- m 00 — «n •^ o r<^ VO O Tt «/^ •^ VO t^ t^ CO 'T •^ ir> VO VO 000 000 o >. 000 000 o o o o d d d d d d d d OHCS CM -^ C/1 cd O £ jn CO 0) TJ- 00 ^ ON 00 Ov t^ r>- CN ON rl- r- >. •^ ?; _> •^ ^ '^ <^ VO 00 r^ c ir» r>- VO "O •«-> 000 000 ta a c 03 ^. o. o. in o. 0. ° 5^ CM '4-* d d d en rt «> O O O fS d d d 5 O O O r^ o •*-*, c 6 6 c O ""« > CQ 3 CQ O u, .-a r^ C^ VO 00 r-l O r~ ON 00 o -- ^ ^ VO >n 000 ^^ 000 000 o o o d d d d d d •a^ pI d d d o; O CO § feb c O '—V •a 5 fsOi-J O (N VO — f~ ov en VO t^ VO m Tj- r«^ 10 VO VO 000 000 o o I 000 000 o o o d d d CO ?? d d d d d d > u

^•^ a, CO o 60 X) 3 O C/3

CO CQ H 6^S 2 Results

1. Leaf nitrate reductase activity (NRA) Application of TPPW proved superior over GW (Table 18; Fig. 7a). An increase of 11.24%, 11.26% and 12.39% was noted by TPPW over GW at vegetative, flowering and fruiting stages respectively. Fly ash concentration, FAio recorded an increase of 28.22%, 27.82% and 27.07% over control at vegetative, flowering and fruiting stages respectively. NRA activity also decreased with increase in fly ash concentrations. TPPW^FAio proved best in comparison to other combinations. In this parameter also GW^FAio was closer towards the TPPWxFAio at all sampling stages, whereas minimum values were observed by GWxFAo. TPPWxFAio registered an increase of 43.34%, 43.92% and 44.24% over control. For this parameter all the interactions proved effective over control at all stages. A consistent decrease in enzyme activity was observed with the increasing age of the plants. 2. Total chlorophyll content Application of TPPW proved superior over GW at all the three growth stages (Table 19; Fig. 7b). An increase of 4.68%, 8.63% and 8.88% was noted over GW. At all growth stages FAio proved beneficial while decreasing value were noted with the increase in fly ash levels. Thus, FA40 recorded the lowest value. FAio registered an increase of 20.56%, 20.68% and 21.06% over control at vegetative, flowering and fruiting stages respectively. Total chlorophyll contents decreased with the increasing age of plants. 3. Leaf N, P and K contents Nitrogen Application of TPPW proved superior over GW, whereas GW recorded significantly minimum value at all sampling stages (Table 20; Fig. 8a). TPPW showed an increase of 15.47%, 18.13% and 20.10%o over GW at respective stages of sampling. Among various fly ash concentrations, FAio proved best over control. As far as interaction effect was concerned, TPPWxFAio proved best in comparison to other combinations at all sampling stages. Significantly minimum value was recorded by GWxFAo, which was found to be at par with GWXFA40. TPPWxFAio showed an increase of 32.79%, 36.21% and 38.74% over control at respective stages of sampling. All the combinations of TPPW-fly ash proved superior over GW-fly ash

63 J3 00 r-- in ON o 00 •>3- r~ ON cs NO o ON o ^ 00 rW 00 cs NO r- O CO CO ON VO O t^ CO O O O '-•-;—' ^ >n vo 60 o o o o o o o a •4-*

ON 00 ON t^ NO (S 60^ 6 C -^ 0\ 10 r«^ cs 00 m 'u cs '^ •—I ON CO ^- n- a CN 'cr rW a\ vo r-' U ^ NO NO O CO ^ C3N .2 m r- »n cs 00 in C3 "w rj- -^ "5- in m m Q o o o o o o o o en a *-> o o o CM n, a CO NO > !r «n NO NO u 3 CO o o o o o o o o &3 60^ •0o) w f 0\ CO -^ O 00 CJN to o tS W-^ Tf ON cs m i-[ k> •4-< ON rW —< O TT K 60 NO m O c V CM ON VO v 'c NO r~ r~- 50 <4_ W-) ID «n O rt J3 u •*-» •+-> O c c<3 00 vo r~ —' O OQ u c J5 J3 o C/3 cs cs r- TJ- CO CO X o O m' •—' CN 00 m •g fe OQ Jg O O OQ CO r^ in m m m •g ^ X >^ X X X en • ^ < UH < DQ < CLi t*^ •a H, o m 13 k> V en > C3 CO X) ^ •*-• C •4-u* cn C 3 ^ CO ON O CNl NO CJN in O *i ON NO '^. ^. «n ON (N <3N C •3 •^ in Tj-' m' -^ CS at JS cs r>- CO O r^ cs CO — *-> >n m NO t^ ro •^ t~~ CO "S, •^ & c 60 ^ ^ ro ^ cs '-' CO cs •<1- TT o o cs CS 00 >n — cs t^ o Q< 60

^•*- • ^» a '••aa "iKsA CS •^ CO 00 Ov rr 00 "O cs TT CO c m 00 cs .w NO (N ON o CO cs CO scd NO CO O 60 m' 00 NO' CO ON NO 00 cs v0n0 cVs) 00 cs o -^ m CO NO NO N•O rr «n m ? m m >n CO *- .S o 6 o= oI ^^a^ c QQ Wi V CQ cs •^ 00 00 ^-« »n NO — TJ- CO O NO 00 in ON 00 to NO cs C3N 13 t^ r-~ r~ NO ON r- CO •«*•' CJN NO r~-' NO CO CO r~-' o cs r~- CO CO CO 00 •^ -^ •^ rs u O cs ON NO •^ (N 00 a> Q. •a NO NO VO r^ 00 t~- •* •<4- -^ m NO «n m c .c o M 3 u o C4-I C c. 0 eso NO 00 cs r- ON CO CO ON —« ON — O in > u->' -^ NO TJ- ^ ON NO CO JS o CO r- CS — cs Si ^ o ON" CO ^ NO' in —«' CO t^' O *~* 00 CO — Tf <7N r- « ?> •^ m in ^r co' co' CO CO CO CO 00 NO 60 •^ -^ •>* 00 rr ^ *-• T m NO NO UQ ^ o 00 en 3 O C/2 CS 1000 n CD. at 5% Vegetative Flowering Fruiting 900 Water 0.134 0.117 0.111 Fly ash 0.190 0.166 0.157 .^ Interaction 0.269 0.235 0.222 '> 800 '•^-t "5 700 4> Vi x: •0 600 s3 p •o ^ S 500 au (4-1 S "bo400 •a la 6 300 _u) 1? 200 100 4

CD. at 5% 2.5 n Vegetative Flowering Fruiting Water 0.012 0.016 0.013 Fly ash 0.017 0.022 0.018 Interaction NS NS NS g 2.0 /—N 0c .£:sP % § 1.5 a, 2 1 tS 2 73 "60 1.0 "a 2P 0 E H 0 0.5 4

0.0 o^^ ^^VVV ^"^^ ^V^^^/ ^"^^ ^•t?" J^ jS^ ^^'^ (Experiment I) Fig. 7. Showing the effect of wastewater and fly ash on BG-256. The data is presented in Tables 18«&19. 0 in ON in 00 cs VO >n /-) o O VO 00 -o c r~ 00 r^ VO 0\ CS > r<^ rr Tf I/O VO VO (S 00 ro VO c O O C/3 —' '— C/3 C/3 c ^ r- ^ fS Id o oo^ <=>. P ;z; <=>. P z Z o 00 00 o X! o, T-H 1-^ VO o\ cs ^ CS «0 00 •^ ON CS o o >r> so w^ r- 00 00 X! o

O GO o — rt 0\ r-H O \o — o\ c 2^ ON O «/^ Tf r- o •ffl. •"^ in m o §^ cs rr fo ^ fe CQ ^ 0 CJ OQ g ^ X >, X X X u O 0o0 to o > ^ < E < « < •o "to CO CO X5

5 «3 s 3 00 00 o\ 00 00 m Tf 00 >—1 00 00 VO 00 ON "^ CO IT) m VO r- CO

60

— fO tS •* ON {S m 00 -- •^ m o O cS ""d- o r-- 00 m

CO

ON -— w-i (N -H (S oq r- Ov ro CO > — ON WO O 00 Tt o t^ 00 ON rf in «n «r> c (S ro ro « (SO t^ r^ t^ 60 5 C4-1 c C30^ o '% CQ CQ o c CO ON r*1 — m 00 ^- 00 r- 00 vo 00 CNl 3 eO O (^ VO (N ON 00 Tf Tt C~- -H T3 to ^ VO r^ r^ 00 ON ON ^ m m e CO *U^ a, -§C CO CO o .Si *-• CO (N TJ- r«^ VO ro O O ON CO ro ON VO r- rr — r-- m r- o ON 00 •* -^ TT in vO VO o — ON O Tf O

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c^ ,>^ »:5^ ^J-'^ Vegetative Flowering Fruiting (Experiment I) Fig. 8. Showing the effect of wastewater and fly ash on BG-256. The data is presented in Tables 20,21&22. Results

combinations. But at vegetative stage on one hand TPPWxFAo was at par with TPPWXFA40 on the other hand it was at par with GWxFAio. This proved the superiority of FAio concentration over other concentrations when interacted with TPPW. Leaf nitrogen content marked a gradual decrease from vegetative to fruiting stage. Phosphorus .'application of TPPW proved superior over GW at all sampling stages (Table 21; Fig. 8b). GW recorded significantly lowest value at all growth stages. TPPW registered an increase of 11.34%, 14.68% and 16.38% over control at vegetative, flowering and fruiting stages respectively. Among various fly ash concentrations, FAio proved best in accumulating the phosphorus at all sampling stages and lowest value was recorded by FAo being at par with FA40. FAio secured an increase of 22.78%, 22.02% and 22.47% over control at respective stages of sampling. Continuous decrease in leaf phosphorus content from vegetative to fruiting stage was observed. Potassium Treatment of TPPW proved efficient over GW, whereas significantly minimum value was recorded by GW at all growth stages (Table 22; Fig. 8c). An increase of 9.61%, 9.63% and 9.55% was reported by TPPW over control at respective stages of sampling. Application of FAio fly ash concentration proved best in accumulating potassium at all stages of sampling followed by FA20. Significantly lowest potassium content was recorded by FAo at different growth stages and it was found to be at par with FA40 at fruiting stage. FAio registered an increase of 20.58%, 20.55% and 20.06% over control at successive stages of sampling. Among the various combinations, TPPWxpAio proved best at three growth stages, followed by GWxFAio. Significantly lowest value was recorded by GWxFAo being at par with GWXFA40. TPPWxFAio secured an increase of 31.89%, 31.95% and 31.78% over control at successive stages of sampling. Like leaf nitrogen and phosphorus contents, leaf potassium content marked a gradual decrease from vegetative to fruiting stage. Among three macro-nutrients, potassium was accumulated more followed by nitrogen and phosphorus.

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3. Yield and quality parameters The yield attributes namely 100 seed weight, pods plant'', seed yield plant'', biomass plant"', harvest index and seed protein content were studied at harvest (Tables 23-25; Fig. 9). The significant data are briefly described in the following pages: 1. 100 seed weight TPPW was found to be better than GW in increasing the test weight of seeds and recorded an increase of 2.17% over GW which gave significantly minimum value (Table 23; Fig. 9a). Among various concentrations of fly ash, FAio performed better in increasing the test weight of seeds. FAo and FA40 gave minimum values. FAio gave an increase of 7.44% over control (FAo). 2. Pods plant"* Application of TPPW proved superior over GW for producing the higher number of pods, and recorded an increase of 36.04% over the later (Table 23; Fig. 9b). Fly ash concentration, FAio proved best in comparison to other fly ash concentrations recording 38.89% more than control. FAo gave the least number of pods which was found to be at par with FA40. 3. Seed yield plant"' Application of TPPW proved superior in enhancing the seed yield over GW and recorded an increase of 25.45% (Table 23; Fig. 9b). Among various levels of fly ash, FAio proved best followed by FA20. FAo and FA40 were statistically at par in their effect indicating deleterious effect of higher doses. An increase of 31.13% was registered with FAio over FAo. Regarding interaction effect TPPWxpAio proved best among various combinations, whereas lowest values were noted with GWxFAo and GWXFA40 which were statistically at par. TPPW^FAio gave an increase of 64.77% over GWxFAo. It may be noted that increase in seed yield was more when TPPW was applied with FAio as compared to FAo, FA20 and FA40. It may also be noted that increase in fly ash concentration consistently decreased the seed yield irrespective of the two waters. 4. Biomass plant"' TPPW showed comparatively better results than GW for this parameter and

65 00 c m CN •^ 0\ 0 CO 00 CJN ^"^ "^. *^ •a TJ CN 00 m t-- VO r- VO (U ^3 m t^ ON ^" CS -H CN ro -ta- -^ VO '•-' 0) :s -H CN O "S 0 0 0 0000 cO '>. W •>% 0 0 0 0 0 0 0 "O CO (A o r~ 00 ro y—^ r-> t-- o lO "O ^ o o r~- o ON in

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\^V •*-> rt ^ 3 •-. CM CN J= JS C tsO "a,?*-. ^ u. O • c^m a U) ^ »r> «o 0\ vo 00 ^ "o 00 O Ov O CN -^ ,^—V o Ov O Ov < o. u> —' CN CN CN CN CN CN CN CN fe- cd JZ c c M E u CO w ,4> c r> >^ o •r) J« cc: fe —< VO m <4-l "O ^ c o o o f] c JS CO r*^ CO o bi) CS CN (N I" 4-^ at •*-> "53 CN CN CN c • ^^ 4-» c •^ ? s O3 o o ^ E o o. VO (N rr TJ- 00 — CO CO ^^Ui ^ k. CN Tf m U bO •a O en CN u •a u J= u •*-ca> /—V C/5 CN CN CN TT Tt TT ^ u "E. J CN CN CN o a. c CO u 1^ CN CN CN > V u 4-t 5 Q, 'eo in *c •C e/5 00 vrJ X) ^ (S 3 o _« 00 CN X5 CO cd H CD. at 5% 100 seed Pods weight plant' Water 0.167 1.564 Fly ash 0.237 2.212 Interaction NS NS

100 seed weight (g) Pods plant' 16 CD. at 5% Seed Biomass 14 Yield Water 0.061 0.066 12 H Fly ash 0.086 0.094 Interaction 0.122 0.133

Seed yield (g plant") Bioinass (g plant')

CD. at 5% Har\'est Seed protein Index content Water 0.064 NS Fly ash 0.090 0.183 Interaction NS NS

O^ ^^ ^^^^ ^^^^ ^v- Harvest index (%) Seed protein content (%) (Experiment 1) Fig. 9. Showing the effect of wastewater and fly ash on BG-256. The data is presented in Tables 23,24«&25. Results registered an increase of 16.30% over GW (Table 24; Fig. 9b). Among various concentrations of fly ash like some other parameters FAio proved best. FAo and FA40 were at par in their values. The optimum concentration of fly ash, FAio gave an increase of 24.76% over control. Among various combinations, TPPWxFAio proved best interaction, while the poorest was GWxFAo although it was at par with GWxFAjo. Significant increase in biomass was noted under TPPW^FAio with an increase of 45.36% over GWxFAo indicating the superiority of FAio as well as TPPW. 5. Harvest Index Application of TPPW proved superior over GW and recorded 7.87% increase (Table 24; Fig. 9c). Significantly minimum value was recorded with GW (Control). Among various levels of fly ash, FAio was found to be responsible for slight increase in the harvest index as compared to other concentrations, followed by FA20. FAo and FA40 gave statistically at par values showing the deleterious effect at this level of fly ash concentration. An increase of 5.11% was recorded by FAio over FAo. 6. Seed protein content Among various fly ash concentrations, FAio also gave, like yield parameters, the optimum value for seed protein content being at par with FA20 and it recorded an increase of 9.95% over FAo. In terms of quality, FA40 proved deleterious as it recorded lower values than obtained by FAo and FA20 (Table 25; Fig. 9c). 4.2 Experiments I & II (Pooled) The design of experiment II was also factorial randomised. The comparative effect of the two sources of irrigation water namely ground water (GW) and thermal power plant wastewater (TPPW) under varied fly ash levels i.e. FAo, FAio, FA20 and FA40 was investigated on another cultivar of chickpea {Cicer ahetinum L.) cv. Avarodhi. Moreover, the pooled analysis of the data of Experiment I and II was undertaken to evaluate the performance of the two varieties together and briefly described below (Tables 6-25). 4.2.1 Growth parameters Twelve growth parameters namely shoot length, shoot fresh and dry weight, leaf number, leaf area, branch number, root length, root fresh and dry weight, nodule

66 Results

number, nodule fresh and dry weight were recorded at vegetative, flowering and fruiting stages (Tables 6-17). 1. Shoot length plant ^ Considering the two cultivars together, TPPW proved beneficial for both i.e. BG-256 (Experiment I) and Avarodhi (Experiment II) as both were equally . responsive for wastewater at all the growth stages (Table 6). Among fly ash treatments, FAio proved best for both the varieties. BG-256 gave an increase of 38.55%, 44.05% and 45.37% over Avarodhi under this treatment, followed by FA20 at all the samplings. FAio with BG-256 gave an increase of 15.39%, 16.07% and 15.40% over control, whereas the same treatments with Avarodhi recorded an increase of 12.15%, 12.75% and 13.82% at three stages of samplings. The value recorded by TPPWxFAio>

67 Results than Avarodhi even under FA20 and FA40 levels of fly ash. It may be noted that shoot dry weight decreased with increasing level of fly ash, and decrease was more with Avarodhi from FAio to FA20, while from FAio to FA40, decrease was almost equal with both the varieties. It indicated that BG-256 was comparatively more resistant than Avarodhi. FAioxBG-256 gave an increase of 29.89%, 32.05% and 31.33% over control, whereas FAio^Avarodhi recorded an increase of 28.81%), 28.62% and 29.32%. TPPWxFAio'

68 Results

6. Branch number plant' Both the varieties performed differently under TPPW treatment as TPPWxBG-256 (Experiment I) gave an increase of 57.91%, 72.73% and 80.31% over TPPWxAvarodhi (Experiment II), while GWxBG-256 recorded an increase of 49.79%, 69.25% and 58.86% over GWxAvarodhi at all growth stages (Table 11). BG-256 grown with fly ash FAio showed 53.21%, 77.03% and 85.95% higher values than Avarodhi at the same level of TPPW and 48.58%, 68.99% and 77.12% at GW treatment. FAioxBG-256 treatment gave an increase of 50.87%, 73.39% and 86.99% over FAIQXAvarodhi. While with BG-256 FAio showed an increase of 53.98%, 51.41% and 54.36% and with Avarodhi, it gave an increase of 55.16%, 43.08% and 44.19% over their respective controls. Branch number also increased with the increase in growth period. 7. Root length plant' TPPW proved effective for both the varieties at all growth stages and the two varieties performed differently under TPPW as it gave an increase of 21.63%, 21.03% and 22.15% over GW with BG-256 (Experiment I), whereas Avarodhi (Experiment II) showed an increase of 18.32%, 17.84% and 17.20% over control (Table 12). It was noted that TPPWxBG-256 gave an increase of 51.60%, 56.40% and 59.51% over TPPWxAvarodhi, and GWxBG-256, 47.47%, 52.27% and 53.05% over GWxAvarodhi. As noted earlier also FAio proved best as the combination FAioxBG-256 recorded an increase of 53.60%, 58.62% and 61.26% over FAiflXAvarodhi. It was also noted that BG-256 grown with fly ash treatment, FAio showed 55.42%, 60.56% and 63.99% and 51.57%, 56.31% and 58.14% higher values than Avarodhi at the same level of TPPW and GW respectively. FAio treatment, with BG-256 gave an increase of 35.62%, 36.67% and 36.04%, whereas, with Avarodhi, it recorded an increase of 28.63%, 29.02% and 27.22% over their control. Root length increased from vegetative to fruiting stage. 8. Root fresh weight plant' TPPW was found to be more effective than GW with both varieties at all sampling stages. With BG-256 (Experiment I), TPPW gave an increase of 18.25%, 19.13% and 18.77%, whereas with Avarodhi (Experiment II), it showed an increase of

69 Results

16.75%, 17.77% and 17.82% over their control i.e. GW (Table 13). TPPW>=BG-256 gave an increase of 36.93%, 40.40% and 44.77% over TPPWxAvarodhi, while GWxBG-256 secured an increase of 35.19%, 38.80% and 43.62% over GWxAvarodhi. FAio proved best thus FAioxBG-256 obtained an increase of 38.95%, 41.35% and 45.52% over FAioxAvarodhi. It was also noted that BG-256 grown with fly ash treatment FAjo showed 39.62%, 41.94% and 46.06% higher values than Avarodhi at the same level of TPPW and 38.21%, 40.78% and 44.92% at GW treatment. With BG-256, FAjo gave an increase of 23.76%, 24.75% and 24.98% whereas, with Avarodhi 19.67%, 21.84% and 22.47% over controls. Root fresh weight also showed a consistent increase with increasing age of the plants. 9. Root dry weight plant'^ TPPW proved beneficial in enhancing the root dry weight for both the varieties at all the three stages of growth. TPPW, with BG-256 showed an increase of 53.28%, 52.94% and 52.68%, whereas with Avarodhi it gave an increase of 35.16%, 36.75% and 39.55% over control (Table 14). It was noted that TPPWxBG-256 recorded an increase of 52.03%, 60.35% and 67.10% over TPPWxAvarodhi, while GWxBG-256 registered an increase of 34.07%, 43.37% and 52.73% over GWxAvarodhi at all growth stages. BG-256 performed better than Avarodhi as it gave higher values in most cases. It also proved superior over Avarodhi under different levels of fly ash treatments. It may be noted that root dry weight showed decreasing trend with increasing concentration of fly ash. FAio proved best in enhancing the accumulation of dry matter for both the varieties and under this treatment BG-256 recorded an increase of 45.59%, 53.57% and 61.95% over Avarodhi. BG-256 grown with FAio gave 53.85%, 61.17% and 68.86% higher values than Avarodhi at the same level of TPPW and 34.48%, 43.87% and 52.84% when grown under GW. It was also noted that FAio with BG-256 recorded an increase of 50.00%, 50.58% and 50.82% whereas with Avarodhi, 47.83%, 50.00% and 50.00% over control. Root dry weight showed a consistent increase from vegetative to fruiting stage. 10. Nodule number plant' TPPW proved beneficial and effective in increasing the nodule number for

70 Results both the varieties at all sampling stages (Table 15). BG-256 was superior in producing nodule when compared to Avarodhi under waste water as well as GW indicating its superiority in possibly higher nitrogen fixation. This variety also proved superior over Avarodhi when compared under different levels of fly ash. Although fly ash treatment FAio proved optimum for both varieties, however, BG-256 was more responsive showing an increase of 64.17%, 73.53% and 66.39% over Avarodhi at vegetative flowering and fruiting stages respectively. Similarly, BG-256 responded better than Avarodhi under FA20 and FA40. It may be important to note that nodule number decreased with the increase in fly ash levels in both varieties. The value obtained by FAioxBG-256 was found to be highest among all the values. Number of nodules increased from vegetative to floweringstag e and started decreasing from flowering to fruiting stage. 11. Nodule fresh weight plant'^ TPPW was more effective in increasing the nodule fresh weight than GW. Both the varieties were almost equally responsive with TPPW treatment (Table 16). FAio proved best for both the varieties and BG-256 gave an increase of 62.14%, 69.93% and 53.36% over Avarodhi due to this treatment at vegetative, flowering and fruiting stages respectively. FAio with BG-256 showed an increase of 30.46%, 31.12% and 33.21%, while with Avarodhi 27.23%, 28.92% and 30.05% over FAo. Nodule fresh weight also increased upto flowering stage, after that it started decreasing upto fruiting stage. 12. Nodule dry weight plant'' TPPW proved beneficial in enhancing the dry matter for both the varieties. Wastewater alongwith BG-256 (Experiment I) recorded an increase of 30.43%, 37.35% and 26.32% over control, whereas TPPW^Avarodhi (Experiment II) showed an increase of 25.35%, 25.38% and 25.00% (Table 17). BG-256 responded better than Avarodhi as it gave higher values for nodule dry weight under TPPW as well as GW. Regarding fly ash treatment, FAio proved best for the same and FAioxBG-256 gave an increase of 53.03%, 53.72% and 57.41% over FAio^Avarodhi at successive stages of sampling. Similariy, BG-256 gave better values than Avarodhi even under control, FA20 and FA40 levels of fly ash. It may be noted that with increasing levels of fly ash

71 Results nodule dry weight started decreasing for both the varieties. Nodule dry weight increased upto flowering stage then it started decreasing. 4.2.2 Physiological parameters Physiological observations namely leaf nitrate reductase activity total chlorophyll contents and NPK were investigated at vegetative, flowering and fruiting stages. Most of the data were found significant and described briefly as follows (Tables 18-22). 1. Leaf nitrate reductase activity (NRA) Waste water enhanced the enzymatic activity in both the varieties. Therefore, TPPW with BG-256 gave an increase of 11.24%, 11.26% and 12.39% whereas Avarodhi recorded an increase of 9.67%, 10.32% and 13.84% over respective controls at vegetative, flowering and fruiting stages (Table 18). TPPWxBG-256 showed an increase of 22.59%, 25.59% and 25.75% over TPPWxAvarodhi, whereas GWxBG-256 reported an increase of 20.85%, 24.40%) and 27.37% over GWx Avarodhi. BG-256 gave better results than Avarodhi under waste water as well as ground water. FAio proved optimum for the two varieties tested and BG-256 recorded an increase of 23.08%i, 26.04% and 26.92% over Avarodhi because of this treatment. BG-256 grown with fly ash treatment (FAio) showed 24.06%, 24.08% and 25.56% higher NRA than Avarodhi at the same level of TPPW and 21.98%, 24.87% and 28.53% at GW treatment. FAio with BG-256, gave an increase of 28.22%, 27.82% and 27.07% while with Avarodhi, it showed an increase of 25.02%, 24.27% and 24.08% over control. It may also be pointed out that BG-256 responded better than Avarodhi under different levels of fly ash. Enzyme activity decreased with increasing level of fly ash and decrease was more pronounced in Avarodhi. Leaf NRA decreased with increasing age of the plants. 2. Total chlorophyll content TPPW effectively enhanced the chlorophyll contents in both varieties (BG- 256; Experiment I and Avarodhi; Experiment 11) and their response was on similar lines under waste water treatment at the three samplings (Table 19). Undertaken in these trials, fly ash treatment FAio proved best for both varieties as BG-256 recorded an increase of 12.78%, 14.82% and 14.72% over Avarodhi because of this fly ash

72 ^w^* Results

concentration. On the other hand, FAio with BG-256, registered an increase of 20.56%, 20.68% and 21.06% whereas with Avarodhi, it gave an increase of 17.63%, 17.48% and 18.21% over control. Total chlorophyll content also decreased with increasing age of the plant. 3. Leaf N, P and K contents Nitrogen Like other parameters discussed so far here also TPPW was found to be superior to GW in enhancing the leaf nitrogen content in both the varieties. TPPW, with BG-256, showed an increase of 15.48%, 18.13% and 20.10% whereas, with Avarodhi, it gave an increase of 12.61%, 13.64% and 17.30% over controls (Table 20). It was noted that TPPWxBG-256 recorded an increase of 5.19%), 7.38% and 5.69% over TPPWxAvarodhi, while GWxBG-256 registered an increase of 2.58%, 3.31% and 3.23% over GWxAvarodhi. BG-256 responded better than Avarodhi as it gave higher values under both waters. It was noted that TPPWxFAioxBG-256 recorded an increase of 4.35%, 6.69% and 4.92% over TPPWxFAioxAvarodhi, whereas, GWxFAioxBG-256 registered an increase of 2.61%, 4.20% and 4.17% over GWxFAioxAvarodhi at vegetative, flowering and fruiting stages respectively. BG-256 also gave better results than Avarodhi under different levels of fly ash. Both the varieties were found to be equally responsive to FAio treatment, as it gave similar increase over its control at all the sampling stages. Leaf nitrogen content decreased with increasing age of the plant. Phosphorus TPPW effectively increased the phosphorus content and the variety BG-256 grown under TPPW showed an increase of 11.34%, 14.68% and 16.38% over GW, whereas, Avarodhi, under the same water treatment showed an increase of 10.80%), 11.22% and 11.39% over GW, at vegetative, flowering and fruiting stages respectively (Table 21). TPPWxBG-256 recorded an increase of 6.69%,, 9.65% and 17.05% over TPPWxAvarodhi, whereas, GWxBG-256 gave an increase of 6.17%, 6.34% and 12.03% over GWxAvarodhi. FAio treatment proved best and BG-256 gave an increase of 8.07%, 9.02% and 16.58% over Avarodhi due to this concentration of fly ash. BG-256 grown with the same fly ash treatment showed 8.42%, 10.89% and

73 Results

18.18% higher phosphonis content at the same level of TPPW and 7.42%, 7.39% and 14.77% at GW treatment. Leaf phosphorus content showed a decreasing trend with increasing age of the plants. Potassium Like N and P, TPPW also proved beneficial as it increased leaf potassium content. Both the varieties were equally responsive under TPPW treatment (Table 22). FAio proved best and FAioxBG-256 gave an increase of 6.77%, 7.80% and 8.28% over FAio^Avarodhi. Similarly, FAio with BG-256 showed an increase of 20.58%, 20.55% and 20.06% whereas with Avarodhi, it gave an increase of 18.60%, 18.58% and 17.65% over control. Leaf potassium content also showed a decreasing trend with increasing age of the plants. It was also observed that leaf potassium content was accumulated more by the leaf tissue followed by nitrogen and phosphorus at all the three stages of growth. 4.2.4 Yield and quality parameters The yield and quality characteristics studied at harvest included 100 seed weight, pods plant'', seed yield plant'', biomass plant'', harvest index, seed protein content and seeds pod"' (Tables 23-25). The significant data was considered in the following pages. 1. 100 seed weight TPPW proved beneficial for both the varieties. In case of BG-256, TPPW showed an increase of 2.29% whereas, with Avarodhi, it gave an increase of 6.40% over its control (Table 23). Regarding fly ash concentrations FAio proved best for both the varieties therefore, FAioxBG-256 gave an increase of 20.34% over FAio^Avarodhi. BG-256 FAio recorded an increase of 7.44%, whereas, with Avarodhi, it gave an increase of 4.64% over control. With different levels of fly ash BG-256 gave better values than Avarodhi, even FAo, FA20 and FA40 gave higher values with this variety. 100 seed weight started decreasing with increasing levels of fly ash. 2. Pods plant'* TPPW increased the pod production in both varieties. TPPW, with variety BG-256 (Experiment I) gave an increase of 36.04%, while with Avarodhi

74 Results

(Experiment II) it gave an increase of 33.43% over control (Table 23). TPPWxBG-256 performed better and gave an increase of 50.61% over TPPWxAvarodhi, whereas GWxBG-256 recorded an increase of 47.73%, over GWx Avarodhi. Regarding fly ash treatment, FAio enhanced maximum pod number as it gave (FioxBG-256) an increase of 51.08% over FAio Avarodhi showed 52.12%) increase at the same level of TPPW and 49.60% at GW treatment. Both the varieties were found to be more responsive for FAio treatment. 3. Seed yield plant"^ TPPW proved more effective than GW as it increased seed yield plant"' with both the varieties. BG-256 (Experiment I) treated with TPPW gave an increase of 25.45%, whereas, vAth Avarodhi (Experiment II), it showed an increase of 24.71%) over its control (Table 23). It was noted that TPPWxBG-256 gave an increase of 92.38% over TPPWxAvarodhi, while GWxBG-256 recorded an increase of 91.25% over GWxAvarodhi. FAio proved best as BG-256 gave significant increase of 93.68% over Avarodhi under this concentration of fly ash. BG-256 grown with FAio produced 94.07% higher seed yield than Avarodhi at the same level of TPPW and 93.49% at GW treatment. It was also noted that FAio, for BG-256 increased seed yield by 31.13%), whereas for Avarodhi, it recorded an increase of 28.89%) over control. Thus, it may be concluded that TPPW with FAio increased seed yield in both the varieties. It may also be concluded that BG-256 responded better than Avarodhi at different levels of fly ash treatment. Seed yield plant"' decreased with increasing level of fly ash and decrease was more pronounced in Avarodhi. 4. Biomass plant'^ TPPW as a whole increased the biomass of plants in both the varieties. The waste water was almost effective for both the varieties (Table 24). FAio proved best and the combination FAioxBG-256 gave an increase of 60.15%) over FAio^ Avarodhi. BG-256 FAio gave an increase of 24.76%), whereas Avarodhi gave an increase of 23.10% over no fly ash control. 5. Harvest Index TPPW effectively increased harvest index for both the varieties (BG-256 and Avarodhi). The two varieties responded on the same pattern when treated with TPPW

75 Results

(Table 24). Regarding fly ash treatments, FAjo gave the maximum harvest index as this treatment with BG-256 recorded an increase of 20.98% over Avarodhi. Thus, combination (FAio^BG-256) recorded an increase of 5.11%, whereas, with Avarodhi, it gave an increase of 4.56% over FAo. 6. Seed protein contents Total seed protein contents was not affected under TPPW treatment, which can also be a positive point for the waste water application to leguminous crops specially the chickpea in the present study. It may be pointed out that aljleast quality of the seeds was not adversely affected. But fly ash concentration, FAio gave optimum value being at par with FA20 (Table 25). Therefore, FAioxBG-256 gave an increase of 13.45% over FAio>

76 flowering and fniiting stages respectively (Table 26; Fig. 10^^ Aiti^"ii^"jyi nitrogen treatments, Nio proved optimum in enhancing shoot length, being at par with N20. Comparatively higher dose of nitrogen i.e. N30 gave lower value at all the sampling stages. An increase of 13.73%, 14.77% and 14.12% was obtained under fertilizer dose of Nio over control. A linear increase in shoot length was observed from vegetative to fruiting stage. 2. Shoot fresh weight planf^ Application of TPPW proved beneficial and gave an increase of 22.63%, 22.04% and 22.01% over GW (Table 27; Fig. 10b). Nio proved optimum at all the stages and N20 was at luxury consumption as it did not increase the fresh weight. The lowest values were recorded by No at all growth stages. Nio recorded an increase of 28.78%, 32.39% and 29.06% over control. Among interactions, TPPW^Nio proved optimum at flowering and fruiting stages of sampling being at par with TPPWxNao followed by TPPW^Nao, whereas lowest values were recorded by GW^No. Nitrogen treatment of 10kg ha'' proved optimum when interacted with TPPW as well as GW. TPPW-nitrogen combination gave better results than GW-nitrogen combinations. Shoot fresh weight increased linearly from vegetative to fruiting stage. 3. Shoot dry weight plant'* TPPW proved more effective for dry matter accumulation and gave an increase of 19.78%, 18.74% and 19.93% over GW (Table 28; Fig. 10c). Nio proved optimum at all the three growth stages. N20 was at par and it was followed by N30, whereas lowest value was recorded by No. An increase of 38.08%, 41.64% and 40.75%) was given by Nio over No. The data was significant only at fruiting stage where TPPWxNjo proved optimum being at par with TPPWXN20, followed by TPPWXN30. Similarly GW^Nio proved optimum among ground water combinations. TPPW-nitrogen combination gave comparatively better results than GW-nitrogen. Thus, it may be pointed out that TPPW proved beneficial in terms of fertilizer economy. Shoot dry weight increased consistently from vegetative to the fruiting stage of growth. 4. Leaf number plant'' TPPW proved efficient for increasing the leaf number and showed an increase

77 VO 00 lO VO 60 .S r> TJ- oo' in 'C O r- c u ^ Tt •^ >n •<:}• c/2 on Q. VO T3 ro •^ •^ o m m 00 o 2i 2 Q o d d d d —" C U E

i 60 VO CO O O o\ r-- fo O VO o\ > *^. °°. ^. \o oo*—<' m r^ 00 r— o. a\ vd fo \o ••§ Tj- ^ ^ ^ O) oo c/3 Vi en Ti" TT M VO VO 03 u O u-i »ri ON 2; 2 IZ; o o 60^ 6 or d d d d > W^ —' 00 00 00 I •^ r- o ON q o V -a ON vd m vd oo' VO VO o m 'S- -^ CO o X) O c o — 00 o —I OO o 00 O c X oo' as • ^ < iz < < H u C3

60

•»-^> co: 5 M ? a, o m 00 r^ m ON C w-i VO Ov .— (U cd •«-* r-' cs 00 iri ts ON" t—I m 80 VO t^ O

^ 60 -T3 C o 2 TJ- m Ov VO O 00 m >o •^ m (^ '^f 00 en >/^ 00 f<^ O VO vq --« Tt VO O vd cs ON r~-' •«*•• ^ -^ (N t-' <^ f«^ •^ •^ ^' 00 in VO r~- VO cs m

•^ > I M VO VO ^ C 4-> 00 m r-- 60^ ON ON 0\ 00 ON •o. '^. so r~ VO r- oo -rt ^ ST) '•5 r<^' ^ r-' V) Tj- NO ed C o r-' TT o •" (S m m «s •i^ •^ o •^ VO r- r~- NO ^^ m Tf Tj- ON VO m CO 60J r~- q 00 \o in VO ti c x> ^1 80 n CD. at 5% Vegetative Flowering Fruiting Water 0.822 0.759 0.740 70 Fly ash 1.163 1.074 Interaction NS

16 CD. at 5% 14 H Vegetative Flowering Fruiting Water 0.207 0.229 0.247 Nitrogen 0.293 0.324 0.350 £ 12 Interaction NS NS 0.495 "S.

c 2

O^ ^^^N'^^^^^^'^ Vegetative Flowering Fruiting (Experiment III) Fig. 10. Showing the effect of wastewater and nitrogen on BG-256. The data is presented in Tables 26,27«fe28 0) 00^ vq —; c IT) vo vo vo ^^ CM 00 CM o Ov ON vo r- ^ c/3 —' CM o CM c^^ -^ vo vo ^ o o o o o o (U a, on o ON —I o ON CM -^ 00 t^ OO »—' m Ov 00 —i CM o o K '-'' ON en CS f<^ m M- vo oo ex —' CM —' ct3 — C>1 CM Tf- >ri UO ^ T3 C Q o o o o o o 03 .60 U

&0^ 00 r^ oo VO (S 'I- O vo •«*• 00 CM O > ja ° vo o r- o t- Du JS Ov •<;f CM « S o CO X) tfa (50 •^ CM 00 r^ ^-< t- O (U ^ CM ^ ^ CM vo •PQ o ia Ov •—' O IT) oo' vo" •H c C ca 3 o c O 00 ^ 2 DH i: bO ^ o

ca V Ov -^ o — CM f,, as t-^ 00 Ov c ^^ O 00 Ov r<^ 00 ON oo' w-i ID ^ CM CM CO O u, .^ C o >/-) m -^ O CJN ^ rj ^^ CO ^ ^ "1 00 o rr r^j °l o f: Ov O Tf C^ o Tf v^' in* r~-' ON 00 VO < ^' ON' t^' Ov >ri CM c vo CM m en

00 00 00 oo n vo Ov vo r<^ ON CM NO > ON m m 00 c PQ en «/^ vo r- r- r~ o ^ ^ NO f. . O «ri vd w-i IT) oo' o Ov 00 CM O ^' 00 Tf ^-' on '-> CM CM CQ 6 O « E^ Ov r~ CM _ CM CO o. rr 00 2 :::; VI '- m •Si «« CM -* oo VO vo VO r^j rj r- t> CO u-> VO >/^ Ov p O -C t/3 00 o c^ —-' 00 •

§ ^ o •-I i rv| o vo CM — r- CM — r~ -H O ^ Ov c/i — OOi-l cs ^ vo r- n vn r^ ^- t^ o o o ^ 't' -^ vo t~-: t-: ex <*^ r«1 CM CM O -* GO t/3 X) o O -— ^ CM CS 2 Z Q O c O O O O O O CO E 00^ r^ 00 00 r- r- o >o ^ o ^ VO ^ > 00 TT oo m o. c 'C Tf' w-i *r\ 1/1 r^ ON 00 •4-* fn r- r- t^ c/D CO c/3 o u x: o (30^ o q q -. ;2: Z Z o, J2. 6 <*- .SP > o o o o o 'S VJO vn o t<^ r~- v~) in o m o Z T^ TT ON r-' ON 00 rj-' u-i -^^ 13 C/l o CO o X3 »—o ' O ON <0 ON CS VO CQ c ^ rn ON vq CO m 00 c X o d CO CO >r( NO' m' CQ 00 C < 2 " o a m 3 h rt X pk. XXX S-' t« > ^ < iz < CQ < OH U CO

00 I s to 60 > '" ON O c VO O r- CM ON ro O m a CM IT\ NO' OO' 00 c *-• o "?« « §0 c •^ »n o r^ — rj- r- rr VO VO VO VO ^ 2 vq o en 00 O ON «0 00 C^ vq CM ON CO a| o vd oo' r-' O r<-i •^ ON CM r-i CM' < < •S > o r—< yr\ r^ t^ ON 00 bO^ 00 rt VO a^ ^ m > ON f^ m ON vq •—' so o vq ri r^ m «n t4_r O tN CO f^ K ON oo' O "^ 00 a «^ 6 Pi 3 T3 > CQ CQ O_ .H^

ON O O CO CM en 00 O •* CO CL. 0) -5 4) CO 00 CN O ON vq f»i vq CM •«*• c^i 00 m o t-^ CN CM' r-' ON 00 cs m' CO CM •^ r^ ^ I < •^ CO c c: C o 00 I-) «/-> t-~ •—I 00 ON ON cn u-i ON m ^ CM •a 4) m VO in vq t--- — CO — o »- «n f<-i 00 Tt NO CS CM CM in vo" VO < 00 O ON aq w ^ •C ,<» to O 00 cs c -3 « s OS CO m H 2 Results

of 37.07%, 44.21% and 39.74% over GW (Table 29; Fig. 11a). While nitrogen dose of 10kg ha"' proved optimum at all stages being at par with N20. An increase of 19.63%, 21.73% and 20.79% was recorded by Nio over control. TPPWxNio proved optimum at flowering and fruiting stages as the data at vegetative stage was non-significant (Table). It was at par with TPPW^Nzo followed by TPPW^Nso and TPPWxNo. Among GW combinations again GWxNio proved optimum. Nitrogen treatment of Nio in combination with TPPW as well as GW gave better results than other combinations. A linear increase in leaf number was observed from vegetative to fruiting stages. 5. Leaf area plant'^ TPPW gave better effect than GW at the three successive stages of sampling giving an increase of 26.17%, 24.83% and 25.32% over GW (Table 30; Fig. 1 lb). Nio treatment proved optimum and it was at par with N20 followed by N30. An increase of 35.34%, 36.35% and 35.49% over control was recorded by the optimum treatment. TPPW-nitrogen combinations gave better results than GW-nitrogen combinations. TPPW xNio recorded an increase of 70.33%, 69.80% and 69.51% over control. However, it may be pointed out that all combinations of nitrogen with TPPW were more effective than their counterparts in GW and nitrogen except the combination of GW with N20 which was better than TPPWxNo showing the importance of nitrogen which should be included whenever TPPW is applied to chickpea. Leaf area increased from vegetative to fruiting stage of growth. 6. Branch number plant'^ Application of TPPW proved effective over GW. It enhanced branching by 35.57%, 33.71% and 27.58% over GW (Table 31; Fig. lie). Nio enhanced branch number optimally at all the three stages of sampling. An increase of 35.81%, 38.92% and 37.21% was registered by it over No. N20 proved wasteful, while N30 proved toxic. TPPWxNio proved optimum for this parameter being at par with TPPWXN20 followed by N30 at all the growth stages. Among GW combinations, GWXN20 gave optimum value being at par with GWxNio and TPPWxNo at vegetative stage but it was found to be at par with GWxNio at flowering and fruiting stages. TPPWxNio gave an increase of 82.27%, 84.22% and 75.77% over control. Branch number showed a

78 c •J CO •^ r- CM r- m K CM' in 00 S; vo m on o OS r~ c/D CO ON m o m ^ CM vo 00 2; ^^ O -^ CM CN CM **» to o O 00 -"^ O — '- CO t^ t^ C-- c ••-* t-~ m o VO o o m CM VO o %> o m vo C m •^ ^ •^ o vo c/D o. w-i o en OS m CO o CM CM o »o vo i^; -o c c r- ^ o\ Q O —' — CM oi CM cO U u o. 00^ o • 15 •-* •>;r -^ CM OS c/3 on 00 O 00 o r< o u ji: CM o. O o < •^-^6 or g r r «9 z 2 z CQ <4-c o c o CM »r> Tj- ON — W-) r» r^ CM cn c-~ o O I—1 00 o CM r- u 00 -I o o -^ < CO x> CO E X) C O (4-1 r-. ^ Tt 00 00 m CO m O t^ o r^ t~ o iff X O '-' vo oo cs c f^ O 00 X X 3

CO x: on

CO

D CO >o — r» o c o •—' VD CM OS so CO oo — o r-~ m »D ^

fe S (U o OS ON Ov CM " CM \0 00 00 0\ Os VD VD VD m > "^. '^. "^ VO 00 00 m ^. vo c*- O ^ g^ vr> vri vri V) vd ^ o^ CJv O sd sd '^ >0 r> vo fS OS vn CM - >D < 6 6 or O « PQ ^^ > E^ 00 r- 00 m -^ TT ON VD — 00 CO {x t^ r^ cs o — ^ VD ^ 4> CO o en r*^ c*^ Ov' sd vd — ^^ CO vrj lo vr^' vd '^ o^ OS >D CM CM r- »D vo o\ r^ •o c2i «o t- vo £ ct3 §^ o vo vo —' 00 0\ ^ CM OS — %/i — O ro f-; ^ vo 60-5 O en CM — VO Tf V CO z oo' E •*-» CO dd CD. at 5% Vegetative Flowering Fruiting Water 9.426 11.146 15.761 Nitrogen 13.330 15.763 22.290 2000 H Interaction 18.851 22.292 31.522

25 n CD. at 5% Vegetative Flowering Fruiting Water 0.099 0.195 0.234 Nitrogen 0.139 0.275 0.330 Interaction 0.197 0.389 0.467

Vegetative Flowering Fruiting (Experiment III) Fig. 11. Showing the effect of wastewater and nitrogen on BG-256. The data is presented in Tables 29,30&31 NO NO o\ J3 O CS ON o NO 00 00 O >< H ^ s; o VO r- S § en (Zi S 00 oo C 1—< ^. ^. ^. ^ 2 <»• ^ O 00 00 JU S — in (A CO •* ON ON VO en b4 ON 00 1—> VO •>!«• o O •^ c T}- t-~ 00 NO 00 m 00 o D- y vd •4-* m r-1 .—I m NO in C/D 00 ON ON in NO CO U 00 ON ^ c^^ c^ :^ d o Q O NO VO 2 d d CO (J o V OH

00 •^ ON O o 00 fo ON NO > r» m t^ VO o o< IS V) 00* »n •4-» •«* en VO VO c« o .2 ON o 00 00 VO VO 00 ir> ON ir> O o O Tt --^ t^ ON 2 o. u*o o E ON 00 u (A o 00 00 00 00 o CM' in" ON •o NO O 00 ON NO c ON "cO JS O «n CO "Q- CO X> «j U u CO ON NO ON NO ON c X 00 O ON ir\ « fe ffl CQ H 1—1 00 c ON o X X 3 o OQ <

CO

cu DO H

to »n oo ON CS O 00 NO 00 C O) CO eq VO NO (S ON CS 00 60 m I—I t^ o •^ en O •^ VD o en «n ON

c ON fS ON ON TT o 00 T}- VO ON ON r- O ON > >r^ (N r- CS VO -rf r- 00 t-~ VO »r> VO o o \r, vn t~~ NO 0} to a> — t—< 1—! r*i -- f--' m' ri 00 ri —' K ON <-• CO CO c NO lO O 00 — •^ *r, »—if 00 00 —^ ON m ^ TT Tj- VO m »—• •^ CS VO —I 00 «i ^ ^^ ^ CS -- "" " 2 -^ If U CO o ^ Tf pn '^ •^ O yn •«1- r^ NO r~- CS m •o « CS CS o O •«»; (N tN TT m r- en r- o ON CO — ^ NO iri ^ •> O O wS vd en c^' ON •'T CS un « CO o > 00 ^ — NO >n O 00 00 TJ- «0 O CS m m 0c>0 O ON ON ^ J= *-• c m •^ •«1- (S m TT « 60 ,<0 ? > J^

to O o r<^ Ji CQ r

C/3 a. on o TT 00 ^ (S r^ m 00 *^ r^ O CS — t> -^ OO ON O •^ _c 00 00 o r-' o\ 00 m' r- »o in ON OO o

c 00^ 0\ ON -^ r*i cs f«^ o c 00 00 r<^ > '^ ^. <^. S© o VO 00 VO oo D. 00 O 0\ •/! •^' 0\ t--" vn vn OS r~- o O O o O oo o o o o o o o a. B BQ 3 > B m ro 00 •^ fO Tf o O ". "*. *^ t^ c^ cs c 00 O OS rr as K

X3 C u o 00 OS ON ^ o —I O VO 00 r>. ^ rt is- c X <2 vo K VO O TT CNJ C t) fc CO 3 •c -«^ X i: X M U >c< 3 ^ < 2 < OQ < u. 60 rt CT3 4-u1 •*-» «J C/3 ^ J3 00

O u 00

4-* <1> o 00 o •a 4-» ON ON •'J- — ^ VO O ^ O — •^ m •^ c "(3 'S cn vo o r-- •^ «/^ m t- r- i^ — ^ VO E <4^ ts rW r<^ T}-' vd w-> 00 O ON r~-' CN C3N i-i o ^ CN — u tn 4-» > c (N ON -^ 00 ON ON (d > ro ro ro •^ m i Z Tj- ^- 00 NO •^ '^. •*. 00 (S O VO Ui m • o 1 (S m' oi •^ wo •^' t^ ON 00 m' r--' in *J .s: o c: 5 00 ^ S.O W S o **- tn O •§ ii ^^2 c/3 a 2 Results

linear increase from vegetative to fruiting stage. 7. Root length plant* TPPW proved superior over GW with an increase of 27.28%, 25.33% and 25.68% over GW at all sampling stages respectively (Table 32; Fig. 12a). Nitrogen treatment 10kg ha'* proved optimum as it v^as at par with N20. N30, on the other hand reduced the root length significantly at all stages of sampling. Root length showed a linear increase from vegetative to fruiting stage. 8. Root fresh weight plant' TPPW proved superior over GW, which gave significantly lowest value at all the growth stages. An increase of 23.96%, 20.04% and 21.06% was recorded by TPPW over GW (Table 33; Fig. 12b). Nitrogen treatment 10kg ha'' proved optimum in increasing the root fresh weight being at par with N20, whereas significantly lowest value was recorded by No although it was at par with N30. In case of root fresh weight, a linear increase was observed from vegetative to fruiting stage. 9. Root dry weight plant"* Watering with TPPW proved beneficial than GW which showed an increase of 58.55%, 55.58 and 55.40% over control (Table 34; Fig. 12c). Nitrogen dose of 10kg ha'' proved optimum at all the three growth stages being equalled by N20 followed by N30 whereas expectedly lowest value was recorded by No. TPPWxNio proved optimum being at par with TPPW^Njo, followed by TPPW^Nso and TPPWxNo. Among the GW combinations, the same dose of nitrogen proved optimum. For this parameter TPPW-nitrogen combinations showed better results than GW-nitrogen combinations. TPPWxNio gave an increase of 125.08% and 120.37% over control at flowering and fruiting stages respectively, while the interaction data of vegetative stage was non-significant. 10. Nodule number plant'* TPPW proved superior over GW and recorded an increase of 33.68%, 30.92% and 32.74%i over GW at vegetative, flowering and fruiting stages respectively (Table 35; Fig. 13a). Treatment Nio proved optimum at all stages of growth. It was at par with N20, whereas N30 decreased the nodulation. Nio gave an increase of 28.26%, 29.22% and 27.44% over control. TPPWxNio proved optimum being at par with

79 0) Is, o o NO r-1 00 r«i ON O VO O CM •"^ r> CM NO CO c/D O en en NO t^ ^ ^ «s o o o" o o

oo O r~ en m ON r-~ 00 6C m f ON Tf rf en o. o rn ON NO" O r^ 00 Tj- r- V3 c/3 rf vo »ri CO CS (S (N ^ TT -"T r- <^. ^ ^ c Q _o O O O O O O

60 C3N ON ON ON Tt (S '- f- en ON ^^ rf CM o. vo 00 K in vb m ON • >-< Tj- en w^ •* ON C/2 C/3 *-> '—I en en NO t~- ^ ^ O c o 6C o o o o o o< n O 0) r~ 00 00 vn o on c a\ r- m ^ o O u-i oo' r-' NO en ON •a cs en cs o 13 o CO X> c U o —I ON ON ON ON CQ O ON r^ < O 3 ex XXX < m < b s «3 60

c« O «< tk

ON •—I «-> V ON 00 r~ en C O VO CNJ Cvj un Tf en ON 00 en IT) 00 60 d. -l-t ^ cs CM en O 0) C > u > 60 •a S O ON NO f*^ 00 en NO r- f- CM f- en c •r> en •S5 ^ (^ CN Tf r-: NO < t^ ri d CM en go ^ cs cs ^« S 60

60^ NO r- r~ > ON ir> (N oo »r> ON wn t- O NO 00 c ON ON Tf r^ ON en S o •o « ri m •^ «/1 VO* ON 00 V) •*-» — — ^ «s O *ri cs K Tf ^ u CN| CM r< B I 60^ o CM en en 3 -O 6 03 O .2 § "ou O ^ 00 00 f^ Ti- c^ en O CM — NO en w-i ON 00 'a; ON *n r-- 00 00 en NO 00 CM ON «ri CS vO ON 00 K Tf —• JC 00 fs w-i •- en 00 NO U5 — •si CM —; '-^ NO c^ 00 p CM VO 2 'a o CM -^ vri CJN r- m yn -^ ^ NO en CM CM CM 2. J= ^3 5 60 S.p •c

CQ z 35 CD. at 5% Vegetative 0.751 1.062 NS

1.0 CD. at 5% 0.9 Vegetative Flowering Fruiting Water 0.012 0.015 0.014 Nitrogen 0.017 0.022 0.020 Interaction NS 0.031 0.028

Vegetative Flowering Fruiting (Experiment III) Fig. 12. Showing the effect of wastewater and nitrogen on BG-256. The data is presented in Tables 32,33&34 ^ ON ^ 00 00 00 CO ON 00 •—I oq CO ON t^ c -^ 00 C/3 i—I C/D 00 CZl o. o ;2; - ;Z 2: Z en s (X o o o 3 o vD ro "n o •^ -^ -^ •4-* VO 00 t~ m —I CO oc (S rf m t-. r- ON '—I u-i vo '^ o •4-* vo 00 r~ ON CO •—I 00 NO CO ex CO m Tj- r-' CS r) C/3 O OO CO 00 o P q ;2 -r Z ;2 Z a, CQ o o o o V «r> vo "O «r^ 00 r~- >n f^ SO r- t^ t^ o CO >n TJ- ON CO •—I •o

1/1 U vo ON CO rf C^ 00 U CO O fS o •—I «/^ .«. c —' CO CN vo ON r^ <2 V c § td X ^ X X X 3

rt

c: CNl rr ON ON cs cs O "^ o r- 00 CS C CO NO o vri ON ^ at) ,- o CN CS o

T3 OO ON 0\ CS CO 00 CO NO NO CO O c ON ON ON o 00 r~ o «o 00 ^ O CQ o 00 O ON r- «n CO ON TJ- fN| O ^ O ^ oi (N <

« C ON 00 Tj- r~ ON 00 00 VO 00 ON ON M J- > ?S °^ 9i >0 CO CO 00 «—I !/-> VO -^ O o^. -. 9 in '^ t^ >i^ Z •o 00 VO ts t^ »n o — — O 'S PQ < oa

o CO ON r- ON ON ON o ON 00 ON CO CX r^ 00 00 0) CO O 'd- «n CO ON ON -I O cs o CO 00 '- O CO t^ r- o CO >r) ON c« -- CNj ON .—i t^ O 00 O 00 •^ ^ CS 00 ON ON — •^ tN «n CO 00 CN O 5J « < O O O '—' CS CS > J^

O 't- co O c -^ ti u s OQ 6^S 2 c \o 10 TT CO in C3N ••-J CO CO vo CO »0 vo CO vo vo cs 0 0 0 cs ^ 0 0 0 0 0 0 0 0 0 0 c 2 d d d d d d 0 g HH d .2 o. 'tA ^ v> o 0 ON t^ 00 NO f- cs vo 00 en CO 00 00 00 00 >o C3N CO vo 0 cs CO vn CO cs o\ r- 0 4-* 0 0 0 C/2 a. 0 0 0 0 0 0 0 0 0 0 0 0 < d d Z C Q 0 d d d d d d cd 'bO U E C/5 2 ^ 00 0 ••• (S ro 00 _i cs r- cs 0 as 0 M- 0 NO > c c •a cs m vo vo cs vo vo 0 cs CO ex, 0 0 0 C/3 c/] 0 0 0 «s 0 0 d d 0 0 0 0 0 0 0 < 60 z •4-* > d d d d d d 0. J3 6 (H < pa > 0

'S 0 fo TT C3N cs ^ t~- CO 00 ON m ">r ON vo vo 0 fn ro •<3- VO •* -0 0 cs «r> < 0 0 0 0 0 d d 13 •t; b 0 /-~v X> 0 0 s C 0 00 00 CO 0 C3N a u 0 0 00 0 ON X v2 0 (S r>i CO CO CO V cs rr 60 < 0 0 0 0 0 d d c3 fe 0 U 0 CQ c X l-r XXX S^ ••-* 3 0-, >^ < "Z < CQ < <

S •" 60

V 0 •i Ci g ^ 8 c r^ ro (^v o c «o 0 vo •^ c CO fo 00 ON 0 CO •* CJN vo vo ^:§ V •—i *-^ CO CO •«1- »o 00 60 J2 r 0 d 0 d d d d d 0 cu 00 •0 2 0 0 c 0 vo 00 r^ 1-^ CJv o 00 VO 10 < 0 0 0" 0 d d d d d < d d 0 d d d 6 ° b

T3 — 0) 0 ••• r^ CN» ir> CO cs 00 vo 60 0 • «• 00 00 00 10 cs C> CO > 03 V) 00 cs vr> 00 CO c JS 00 rt vo vo cs t~- C3N CO CO *•*-» T3 CO «0 •* ON Sg z •a > 6 > < CQ < CQ 0 «

0 m r~ u-> CO C3N 00 0 0 r- CO 0 vo w 0. iri 0 f- ^— TT r^ cs •^ 00 •* vo ^^ VO ON cs 4J to (S CO CN CO vo •^ ON J= en 0 -" ^ cs

§,-7 1—1 1—1 MHJ ts ON 00 vo CO 0 0 vo 00 r~ CO «o r- en — 1—< iri CO ON 00 ON CO ^ ro Z CO vo •^ « "to •—rrI vo r« 0 •—' '—' ^" cs •—' •—' 0 cs CO CO •^ vo VO • 4-' t« 0 •^" c c m u 4> •3 «^ jj C c c 6 c c c e ^ ^ ^ cd 3 It) ^ ^ 0) ^ HI ^ ^ CQ 0 ^S 0 Ui 0 S S ^ s s H ^ 0 ^ s z; Results

TPPWXN20 followed by TPPWxNao at flowering and fruiting stages and the data at vegetative stage was non-significant. Among GW combinations, GWxNio proved optimum being at par with GWXN20 and TPPWxNo. An increase of 68.85% and 68.56% was registered by TPPWxNio over control. Nodule number increased from vegetative to flowering stage and decreased at the fruiting stage. It may also be pointed out that among the three stages of sampling maximum nodules were found at flowering stage while minimum at fruiting stage. 11. Nodule fresh weight p!ant"^ As noted in earlier parameters, application of TPPW proved beneficial (Table 36; Fig. 13b). Nio proved optimum at all the stages being at par with N20. It was followed by N30, whereas lowest value was given by No. An increase of 29.13%, 29.37% and 29.30% was recorded by Nio over control. Nodule fresh weight increased from vegetative to flowering and decreased from flowering to fruiting stages of growth. 12. Nodule dry weight plant'^ TPPW proved superior for this parameter also and gave an increase of 26.42%, 28.71% and 35.23% in comparison to GW (Table 37; Fig. 13c). Nio proved optimum and recorded an increase of 34.69%, 35.11% and 34.52% over control, whereas N30 proved toxic. Nodule dry weight increased from vegetative to flowering and decreased from flowering to fruiting stages of growth. 4.3.2 Physiological parameters Physiological observations namely leaf nitrate reductase activity, total chlorophyll contents and NPK were investigated at vegetative, flowering and fruiting stages. Most of the data were found significant and described briefly as follows (Tables 38-42; Figs. 14-15). 1. Leaf nitrate reductase activity (NRA) TPPW enhanced the enzymatic activity by 14.85%, 15.89% and 16.57% over GW (Table 38; Fig. 14a). On this parameter Nio registered an increase of 19.29%, 18.31% and 17.45% over control at vegetative, flowering and fruiting stages respectively. N20 could not increase the NRA while it decreased under the treatment N30. TPPWxNio proved optimum at the three stages of sampling being at par with

80 O m 00 m u •^ 00 00 (S ON vo (S 00 CS ON ON ^- CO >—' C/3 ON CN o >n m o CS CO ^ O O O ^ -H' --' !t 00 ON ^ •ca o Q o d o ^' ^' '-• CO O o

CO Tj- r- \o ON <—I «n O •^ •^ •^ •* -"^r •^ > r^ cv) CO CO t^ '- 'C ^ XT, V> f^ o o m m vo c/D Vi o f>i vd •4-* O 00 00 CO vo t-~ ^ OD t^ vo

t- OS CO 0*0 00 in CO CO f*^ Tf fO CO O 3 cs vo •a (^ vo ro '- 13 o en c CO c o VO ' — ON m vo < vo •^ ro r~ o c o r~ ro »n v»^ X «2 •^' 00 r-.' ,— o >^ < iz < CQ < CO 60 aen o bO u •a D Tj- CO ON ON O 00 O vo C 00 O ON •<;*; ^ CO u ON 0\ 00 in o o 60 rNj CO Tj- vo r-' in O (S en vo" ^" Tj- vo O. i-i c 00 ON ON 00 CO vo o cs — 00 ON ON ro CO oo O O O r- t-~ r- O vo CO CO ON oo' Co" oo" •*' vo o •5 « cs CO CO TT m »n

00 ON ON cs ; 00 ^f cs «s r^ U1 -^ r~ -^ ON vo CO •^ CO

M O CO OO f3 i 140 CD. at 5% Vegetative Flowering 1.685 Water 1-631 2.383 120 H Nitrogen 2.307 3.370 Interaction NS 1 100

1.0 1 CD. at 5% Vegetative Flowering 'ruiting 0.9 Water 0.0216 0.0593 Nitrogen 0.0305 0.0838 I 0.8 Interaction NS NS "=9 0.7

.'P^^'^ Vegetative Flowering Fruiting (Experiment III) Fig. 13. Showing the effect of wastewater and nitrogen on BG-256. The data is presented in Tables 35,36«&37 t- ON m -^ (U CS ^ in .—' 60^ TT »0 t^ ON ^^ 00 00 m O —' -^ CS C/D C/2 00 o o o o 'a 0000^ 2 2 (L> d d d d

(X CO CO ^ O vo •«-» o vo r^

60 (S •^ m r- 00 00 C>J O -^ _> cS r-« t^ CS vo 00 O 0\ m r- t-» ON 'C rt »r^ u~, ^ CO CO O. o ^ o •4-t o ^ — rs CO c/D c/: O O O O M u <=>. q q q ;z; 2 2 <-> 0000 ^6 0 o 6 6C^ V (L> ^ o\ c-> en CO J3 \o vo -^ — 00 O O w TT >n »n 00 ON ON T3 (U O O O o o o to CO 60 CO CO x: fe•4-CO ' k> J3 an ••-» ^ ^ u s60 •#-M» V 9i CS r~ NO Tj- ^ o ON NO ca TT 00 CO r^ c ^ 4= CS CS ^ o ro -^ •4-J CS CS •-• Tj- in 60 d d d d d d O c to d d cx c l-l 60 0) 0 c ^ NO CS ON m Tj- o Ol O CO 00 NO r~- CO 0 ^ NO m -^ O NO ^^ «n ^- r- r- CS ex •5 CS CS (N ^ m TT CS CS CS CO ^ • •4-^ r- CS ON ^ m in o NO O 00 ^_ — CS r- c CS CO CS J^ •^ m m c 2 •a CS CO CS J» Tf m rj- C4_ O c 0 0 O "^ CO k> •4-* d d d «s d d d O O O CS d d d u 5 < c '^ i 60^ 6 b > 6 3 •« CD •0 < JC PQ •4-» ;« "^ ^ wo o CO O CS 0 •«r CO ^ >n CO ON CO o. •^ VO ^ ON •^ -"S- ON m ON r- o <—I m fW CS CO CS •n Tj- ^ ^ J= CO d d d d d d d d d d d d •^ CO -a c s60 o x: 3 C o 0 /—N k> J ro ON •— NO «/0 •—I rf CS 00 «n Tj- m NO 60 Tt —I 00 O CO »-' >—' ON m 00 a cO C|-c CS CS CS ro TT CO CS CS CS CO ro ro CS 0 s;: 4-* s: d d d d d d d d d d d d 5 60 U •a ,1> u fcl C 0.0 W "3 O '*-' \o' c CO O ro u v E C/D CO X) R] to PQ H H 2: (U cs o oq o o C/3 C/2 00 00 on GO C/3 d d cS d d 5 a. m en 00 t^ tn CO OS ^^ 03 0\ -^ (N c O o ^ (N (N V(0 .-< 00 CO »/-i O. 'C O O (Zl ^ — C/3 C/3 <+- d d d d d d T3 o 0) o P ;2; o q ;z; Z: C Q CO o o o o 'So u 00^ t~- m o > Tf 00 \0 v-. CN ON VO c -: ^. -1 Cn CN CN (S {/5 C/3 C/a C/3 (/3 C/3 (Z) tn o o o '5 o •^ CS 00 —' t^ -^ a fO 00 VO Vi cs 00 >r) o d d d CN (S (N •o en c3 3 d d d XI •o o c vo m >r) •T) ^ OO '^u^ O o -^ ts VO —1 00 c: c ^ cs ^ s o X o 00 O O 03 C o o o d d d 3 •4-oJ XXX > ^ < Z < OQ < P- tsO rt

^ 2

—I \0 OS Tf OO Os t^ 00 VO fO o o t- •^ OS cs so CO CO 00 O .t! vo 00 r~ o 0\ 00 ON CO 00^ 00 CO so r- o\ 00 ^ cs) <0 CO CN JS »n t^ so rs> so OS o o o J^ o o o O CO ^^ O d o d fs d d d u d d d §•0 6 PQ CQ VO cs -^ so t~- cs o vri O CO W1 O CO r~ o\ 00 wo r~- so C3S CO ^^ o o o — TT CO JS en o o o O — -^ *-• rt ^ I d d d d d d d d d d d d •o g § 00 C2 -^Oi O '-> •O « obJ — m r~ r^ 0\ 00 rr so o —' so ^ en -7 soo ro- voo o00 O— Oos t~ OS 00 SO o § TT wo v^ o o o o d d d d d d o o o d d d 5 bO d d d o 'C

TPPWXN20 followed by TPPW^Nso and GWxNo gave the lowest value for this parameter. It may be pointed out that with both waters, Nio proved effective, whereas all other combinations proved wasteful in terms of fertilizers. It was observed that TPPW successively reduce the use of inorganic fertilizers. The nitrate reductase activity generally decreased with the increasing age of the plants. 2. Total chlorophyll content TPPW proved superior over GW and recorded an increase of 6.80%, 9.88% and 10.83% over GW at successive stages of sampling (Table 39; Fig. 14b). Nio treatment proved optimum and was at par with N20 followed by N30. This treatment enhanced chlorophyll contents by 21.36%, 20.14% and 19.48% over control. Total chlorophyll content showed a linear decrease from vegetative to fruiting stage. 3. Leaf N, P and K contents Nitrogen TPPW proved efficacious over GW and recorded an increase of 21.33%, 23.70% and 24.92% (Table 40; Fig. 15a). Effect of nitrogen was not as distinct as noted in some other parameters. Thus all the nitrogen doses were at par in their effect and no nitrogen (control) was lowest recording critically different value. Among interactions, TPPW^Nio proved optimum regarding nitrogen uptake at all sampling stages being at par with TPPWXN20 and TPPW^Nso, whereas minimum value was recorded by GWxNo. It was noted that Nio proved effective when interacted with both waters and TPPW-nitrogen combinations gave higher values as compared to GW-nitrogen. Leaf nitrogen content decreased with the increasing age of the plants. Phosphorus Application of TPPW enhanced the phosphorus content in the leaves of chickpea as it recorded an increase of 9.42%, 18.91% and 13.89% over GW (Table 41; Fig. 15b). Like nitrogen content, the effect of nitrogen doses on phosphorus content was also the same as the three nitrogen doses were at par in their effect. Like nitrogen, phosphorus content also decreased from vegetative to fruiting stage. Potassium TPPW also enhanced the potassium contents over GW and recorded an increase of 15.04%, 16.98% and 10.24% over control (Table 42; Fig. 15c). Among

81 45 c ON O ON f«^ or J2 C/3 o m TT ^ c r- 00 o r^ o r- •*-> O «o' —; vd *r^ vo vo t^ r- 00 ^ C/3 :s r~ -^ fS fo * ^H ^ •<}• Tj- vo 00 CO J2; — «in vo r~ 00 O CO CO in vd r-~' S 60 fea .2 "53 «5 o vo t-~ cs vo ON 00 vn Of •^ 1—1 00 CO O vo CO _c ON o a. >n ON •^ in •* r- o .—' 00 Tf f<-i oo' »n O ^j (S 00 00 ON O O t/3 VO CS ON ,-1 ^ vo oo R] CO »n m in CO ON j2; T3 < in SO m r- oo r- vo c •4-> Q o •^' •<«•' t--' ON ON ;> O b VI o cd bC o ••• O^ 0\ 0\ m ON •—1 O oa C O PS TT CS 00 y^ O M t^ 00 a> 00 vo '^ m t~ 00 O a. M O 'u ^ o rr K »n" 1/1 ro ts r~' vo ed vo r-- r~ •<;r S S C/3 X/i eg 00 O 00 •^ n in ON — ts '^ S en O U^ « CQ b <>; > O

4> O o "O O en o n VO VO r~ 00 00 t^ en VM f tlH ea C3 *-• ^—s X)

O m >n TT 00 m •—1 (S o ON O O O ° tsO *^. *^. *--'. X oo' «n r-' in 'c (S 00 «r> •<;r -t ON (S U O OQ vo r- vo in «r> m vo XXX 3 OK C OH <+- > ^ < iz < CQ < < H o 7 ^-^ CO rt tt "S J: « > 00 to « ^ c ?~ • •—1 (U C •S! « /-—N CO a C o r- r- Tir c r- — r- ON 5^ ^S •

o CO CO 00 o oo ON ON TJ- CO ON "O "?* v JS \o vo — m 00 •«*• >^"> > •a <^ 00 m y^ CO •<*• ON (S c ON CO VO V© CO 00 m o o 00 (N O" V3 «n (S en" r-' '•2 o o oo' O ON Tj-' (S 00 ON «*- s •t-t u ON 00 M- r< vo O CO CO fc. t- VO -« O 3 t3 > < pa «; CQ O 4> 4J e<3 o vn r- ^o ^ tS CO O o ON TT CS o o O vo «^ Ou 2 OO 00 PO •n 00 o CO •^' Tt O 00 o o Ov t^ r<^ vo ON (S CO r~ in r— (S CS r-> ON •o -^ < VO t^ r^ 00 ON ON 00 < •^ m >n vo r- vo in c JC U. b

T3 a; 00,—s o (N »n Tj- CS ON ^ r~ o ON ON ON ON fS ^^ in c/3 _< •^ -^ TT CO CS CO CO fS CS (S rf ON CS r- o -c 3 ""^ o r- m o oo' 00 oo' •^' zo t-' —' ON CS m' TJ-' ^ ON vo ro (N CS t-~ o '^ 00 •«d- CO ^- r- f^^ 5 00 g 00 b< «n vo vo r~ 00 r~- t-~ < •

CD. at 5% Vegetative Flowering Fruiting Water 0.0186 0.0187 0.0193 Nitrogen 0.0264 0.0265 0.0273 Interaction NS NS NS

^^NV'P^^'^ 0^<{^ ^^N^-?^^^

(Experiment III) Fig. 14. Showing the effect of wastewater and nitrogen on BG-256. The data is presented in Tables 38&39

f^ If o 1—1 vn ro m C^ m •^ CO VO 00 o\ —' CS 00 CS — (N o -—' (zi m (/3 c/D on CTS '53 o o ;2: P 2 2; ?: o o o c?v »r) (N t-~ vo t^ 6q Tf OS fS ON ON ON O \0 t^ t~ 00 O ON in .S CS r- m o ^ csi —• o- O ^ (Z) CO •^ C/D C/3 T3 60 Q _o o. o 12 o o ;^ ^ C 60 O o o o o 6 00 ON 'sr 00 VO CN o on vo «£) vo >a ^ Tt o r-- c a z •o O CS r- O CS c/2 ro ro c/3 c/5 cx c o C^ ON 00 J^ ts cNi ci o. o. :z: o q ;z; ;z; o o < XI o UH > —'' ^ —'

Pi ^ > ^ < iz < ca < X 60

00 in m r--

h B T3 C 00 ON ON ON ro VO r~ rr 00 ^ ^ ^ VO r~ m ^o 1—I rr r- ^ r>- Tj- »n a fs

X 60 « c 00 r~ 00 t- Tf VO 60^ CS ON VO m •^ ON VO CS Ti­ •«1- O CS O ON CjN «/^ ON . <=>. in 2 < > UH ^ 3 "O < ^ CQ O <"

>o ON r~ CS r~- m o —' CS CS ON o »n T^ OO — fO ON ^ 00 00 00 ON O O CS •^ m •^ >n m VO 00 r~ X V] ^ (S tS CS CS CS z •" to

o X ON 00 rj- fO TJ- rf C7N 00 CjN >n ro Ti­ vr> ^3- O t- f- CS ro in ON es NO C3N C o vD t^ f- 00 ON CJN CS CO CS Ti- >n Tt «3 -• Q.q •C f^ M O

^^2 oa 2 en 00 lO m •* mVO m^ o lO 00 «o o\ r- O . 9 ;z: o 2 ;z; 2 o o o S a. _3 o O <— VO ir> fo Ov «0 00 '— c>) ^ VO o JuS r-- (N o 00 t- CS 'C ON Tj- ^ ^^ m o. (S rn m' ts rW rn O CS 0 C/3 -a o Q o o o o o 2 ^ Z c O b o o o" o o O 6q o\ o\ -^ TT VO »0 > C .S m VO O a "C "T3 r- ts o »-i Ov >r) Tt TT >/o Tt r^ cx o 00 vq ts •41^ c p tN rW tn m (U O (N CS •* C/3 «/^ (Z) O o < • o o o o J2; c> Z c < o o o o o a. to o 00 00 m cs 00 /^ ON O 00 -"^ o r~-

X3

c TJ- CO ON 0\ •—I O o »- •^ Cvl t^ 00 m c o CO r~; UO fO O t~-; 60 X ^< z < O 60

60

m /^ o\ ^ -^ 00 w^ ts VO VO = c tn '" « S 2 ^ oo m -- >5 O 00 rj- c z •o VO vn ON , . i-i o O rf 00 U m ^ Tt •^' '^ -^ '•3 o hO ^^ < < A 60^ [IH >es 6 3 ^ O •-5 < s ^ CQ O .2

o TJ- VO O VO m o 1"^ 3 5 — Tt 00 o r^ -^ CS ON VO «n TO gL, 00 vrv — O 00 T A. VO zo m' Tt Tf •^' TT Tt CI T*3 iO C l- 3 60 C o O r-v I- < m m en 00 VO ts on — 00 U^ (S VO fS m a o § ts 00 W-) m o t^ m m VO ^ xa: en m m m m 5 60 0.0 o '•-< o en O -§ B XI CO CQ z CD. at 5% Vegetative Flowering Fruiting Water 0.0196 0.0191 0.0189 Nitrogen 0.0277 0.0270 0.0267 Interaction 0.0391 0.0382 0.0377

I 0.20 "a 0.15 u e:i 0.10

0.00 6.0 n CD. at 5% Vegetative Flowering Fruiting Water 0.04S8 0.0225 0.0208 Nitrogen 0.0647 0.0318 0.0294 Interaction 0.0915 0.0450 0.0415

o^^ ^V^^-^^^' 0^^ Vegetative Flowering Fruiting (Experiment III) Fig. 15. Showing the effect of wastewater and nitrogen on BG-256. The data is presented in Tables 40,41&42 Results the nitrogen treatments Nio proved optimum at all the three stages of growth being at par with N20 and N30 as noted in the case of nitrogen and phosphorus content. TPPWxNio proved optimum at all the three growth stages being at par with

TPPWXN20 and TPPWxNso, whereas least value was registered by GWXNQ. Among GW combinations, GW^Nio proved optimum being at par with GWXN20 and GWXN30 followed by TPPW^No. Leaf potassium content also decreased with increasing sampling days. It was also noted that potassium contents were accumulated more followed by nitrogen and expectedly the lowest contents of phosphorus. 4.3.4 Yield and quality parameters The yield and quality attributes studied at harvest included 100 seed weight, pods plant'*, seed yield plant'', biomass plant'', harvest index and seed protein content. The significant data are described briefly (Tables 43-45; Fig. 16). 1. 100 seed weight Nitrogen treatment Nio proved optimum for 100 seed weight (Table 43; Fig. 16a). Like some other parameters it was also at par with Njo- Treatment Nio showed an increase of 8.96% over No. Effect of water treatment and their interaction with nitrogen was non-significant. 2. Pods plant'* TPPW proved effective for increased pods followed by GW and gave significant increase of 40.28% over GW (Table 43; Fig. 16a).Among the various nitrogen treatments 10kg N ha'' proved optimum, as it recorded maximum number of pods plant"'. It was at par with N20. N30 gave the lowest value among the nitrogen treatments except the control. Nio showed an increase of 36.81% over No. TPPW^Nio proved optimum being at par with TPPWXN20, followed by TPPW^Nao. Among GW combinations, GW^Nio was the optimum treatment. Again TPPW combinations proved more effective when compared with GW combinations. Therefore, the Nio irrespective of water proved optimum. An increase of 90.44% was recorded by TPPWxNio over control. 3. Seed yield plant'' In irrigation water treatment effect of TPPW proved superior over GW, the former being 30.20% more over later (Table 43; Fig. 16b). Among the nitrogen

82 O CO 00 m o O TT ro 00 oq VO ^ VO

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n . <^ Harvest Index (%) Seed protein content (%) (Experiment III) Fig. 16. Showing the effect of wastewater and nitrogen on BG-256. The data is presented in Tables 43,44&45 Results

treatments, Nio proved optimum in enhancing the seed yield. It was at par with N20, followed by N30, which proved wasteful. Treatment Nio recorded an increase of 32.03% in seed yield over control. In interactions, TPPWxNio proved optimum as it was equal to TPPWxNjo and followed by TPPWxNso- On the other hand TPPWxNo gave the least value however, it was at par with GWxNio and GWXN20 followed by GWxNao. Among the combinations of TPPW and GW, lowest value was recorded by GWxNo. An increase of 71.01% was registered by TPPWxNio over GWxNo. It was also noted that lower fertilizer dose applied under TPPW proved beneficial as it saved some inorganic fertilizer. 4. Biomass plant'* TPPW increase the biomass significantly showing an increase of 20.03% over GW (Table 44; Fig. 16b). Among the nitrogen treatments Nio gave the optimum value as it was at par with N20 showing luxury consumption of N. N30 proved toxic as it decreased the biomass. The optimum treatment increased biomass 24.16% over control. Interaction was found to be non-significant. 5. Harvest Index TPPW proved effective for the harvest index and recorded an increase of 8.60% over GW while the data regarding nitrogen treatment and interaction was non-significant (Table 44; Fig. 16c). 6. Seed protein contents TPPW proved beneficial regarding this parameter as it increased 4.49% protein over GW (Table 45: Fig. 16c). The effect of nitrogen doses was not distinct except that these doses were better than control. Therefore, Nio proved most economical treatment as it yielded an increase of 7.33% protein over control. 4.4 Experiments in & IV (Pooled) In this factorial randomized pot experiment IV, comparative effect of two irrigation waters and three basal nitrogen levels was studied on chickpea (C. arietinum L.) cv. Avarodhi. Pooled analysis of the data (BG-256; Experiment III and Avarodhf, Experiment IV) were undertaken to evaluate the comparative performance of two varieties. The growth and physiological parameters were recorded at three growth stages. Yield attributes on the other hand including seed yield and seed protein VO IT) ON t^ O (^ vd o o »ri o o 00 '-<' •^ in 00 en c/3 ON c/D c/3 en IS cd C o CO -^ c; «>. Z *>». 2 Z Z o o o -- t/3 00 O •^ Cd ON (S •4-t CO rn C/3 C/3 C/D O (S (S >0 ;Z; Z Z C 'a- »/^ ^ Q cd c U OQ o o o o

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CN CN CN < _« (S 01 (N 13 w c JC u CO •4-* an c CO c o t^ ON cn 00 m »— >% u VO VO '<-> X) vr> ^^ rn vq C) c: 0 •0 c CS m m' r4 (4-1 C<1 >/-) vd vo 0 •4-» c ^ •53 (N CN CN 1 c<] *-4 CN CN CN *-* CO c •0 s pa 3 a> o. 0 E "H. o -o ^^ CN r~ CO a. r~ >o »— (0 u &Q V 10 VO 0 cNin ov 4J J= C/5 /^ vd >/^ CO CN CN cs CN CN CN V ^ 0 sz C 0 CO •0 en CN 00 — r-~ • ^ •-• 0 .B- CN *0c (*0_ CO Jd u 3 00 to dd z Results

content were assessed at harvest. Only the significant data are briefly described below (Tables 26-45). 4.4.1 Growth parameters Twelve parameters namely shoot length, shoot fresh and dry weight, leaf number, leaf area, branch number, root length, root fresh and dry weight, nodule number, nodule fresh and dry weight were observed (Tables 26-37). 1. Shoot length plant'^ TPPW proved beneficial for both the varieties as it enhanced the shoot length upto the fruiting stage (Table 26). Both the varieties were found more or less equally responsive to wastewater. Among nitrogen treatments, Nio proved optimum, being at par with N20. BG-256 showed an increase of 40.20%, 45.03% and 47.71% over Avarodhi under this treatment at vegetative, flowering and fruiting stages respectively. With BG-256, Nio gave an increase of 13.73%, 14.77% and 14.12%, whereas with Avarodhi, it recorded an increase of 12.30%, 13.37% and 12.03% over respective controls. Shoot length showed a linear increase from vegetative to fruiting stage. 2. Shoot fresh weight plant'' Like shoot length TPPW also proved effective for increasing the fresh weight (Table 27). Both the varieties (BG-256 and Avarodhi) were almost equally responsive. Nitrogen treatment 10kg ha"' was found to be optimum as it was at par with N20, and BG-256 showed an increase of 63.88%, 69.94% and 73.56% over Avarodhi due to optimum treatment. It was noted that Nio, with BG-256, gave an increase of 28.78%, 31.91% and 29.06%, whereas with Avarodhi, it showed an increase of 25.97%, 27.11% and 26.09% over their respective controls. Although both varieties responded well to wastewater but it was BG-256 which showed comparatively more increase in fresh weight than the Avarodhi at the three stages which was more pronounced at fruiting stage. Shoot fresh weight increased with increase in growth of the plant. 3. Shoot dry weight plant"' TPPW effectively increased the dry matter production in both the varieties (Table 28). Therefore, TPPW with BG-256 (Experiment III) gave an increase of

84 Results

19.78%, 18.74% and 19.93% whereas with Avarodhi (Experiment IV) it recorded an increase of 22.89%, 22.03% and 20.35% over GW at vegetative, flowering and fruiting stages respectively. Treatment Nio proved optimum for dry matter accumulation while N20 was luxury and N30 deleterious. BG-256 registered an increase of 60.98%, 67.68% and 60.62% over Avarodhi under optimum treatment at all the samplings. Shoot dry weight also showed consistent increase fi-om vegetative to fruiting stage. 4. Leaf number plant'^ Wastewater application increased the leaf number in both the varieties (Table 29). Therefore, TPPW with BG-256 (Experiment III) gave an increase of 37.07%, 44.21% and 39.74% whereas, with Avarodhi (Experiment IV), it showed an increase of 34.87%, 45.24% and 51.43% over GW. Among nitrogen doses, Nio proved optimum and BG-256 gave an increase of 21.20%, 24.75% and 21.58% over Avarodhi under this treatment. This treatment recorded an increase of 19.63%, 21.73% and 20.79% in BG-256 whereas in Avarodhi, it gave an increase of 15.29%, 16.50% and 15.85% over No at all the samplings. Leaf number was increased upto the fruiting stage. 5. Leaf area plant' With both the varieties, TPPW gave better results than GW at all the three stages studied (Table 30). It gave an increase of 26.17%, 24.83% and 25.32% with BG-256 whereas with Avarodhi, the increase was 23.76%, 24.55% and 21.95% over GW. TPPWxBG-256 gave an increase of 34.09%, 42.96% and 47.66% over TPPWx Avarodhi whereas, GWxBG-256 showed an increase of 31.54%, 42.65% and 43.70% at three respective stages. BG-256 performed better than Avarodhi under TPPW as well as GW at all growth stages. Regarding nitrogen treatments, with BG-256 N20 recorded maximum value, which was closely followed by Nio at vegetative and flowering stages, but at fruiting N20 was at par with Nio, while in case of Avarodhi, Nio was optimum being at par with N20 at vegetative and fruiting stages and at flowering N20 gave the maximum value, which was closely followed by N10. At vegetative stage NioxBG-256 gave an increase of 34.53% over Nio^Avarodhi and N20XBG-256 showed an increase of 34.19% over N20XAvarodhi for this parameter. At

85 Results flowering N2oxBG-256 was 44.49% higher over N2o>

86 Results root fresh weight in both the varieties (BG-256; Experiment III and Avarodhi; Experiment IV). The wastewater recorded an increase of 23.96%, 20.04% and 21.06% in BG-256, whereas in Avarodhi, it increased upto 21.72%, 16.24% and 21.06% over GW at three respective stages (Table 33). It was noted that TPPWxBG-256 registered an increase of 43.98%, 47.62% and 48.15% over TPPWxAvarodhi, whereas GWxBG-256 recorded 41.37%, 42.94% and 48.16% over GWxAvarodhi. The treatment, Nio proved the best while N20, being at par with Nio was at luxury consumption. It may be noted that nitrogen dose, N30 gave less values than Nio and N20 for both varieties, but in case of Avarodhi, it was at par with N20 as well as Nio, whereas in case of BG-256, it gave less values than both the lower doses of nitrogen. It proved that N30 was found to be deleterious. TPPWxNioxBG-256 showed an increase of 45.42%, 50.89% and 52.03% over TPPW^NioxAvarodhi, whereas, GWxNioxBG-256 gave an increase of 46.87%, 45.76% and 52.02%. Root fresh weight also increased upto the last stage. 9. Root dry weight plant* TPPW proved beneficial in increasing the dry matter accumulation in both varieties (Table 34). TPPWxBG-256 registered an increase of 67.21%, 72.68% and 76.33% over TPPWxAvarodhi, whereas, GWxBG-256 recorded an increase of 40.88%, 53.91% and 61.16% over GWxAvarodhi. BG-256 responded well when compared with Avarodhi under both the irrigation waters. This variety gave better results than Avarodhi under wastewater and nitrogen treatments. Nitrogen treatments significantly affected this parameter thus Nio with N20, as it was at par and NioxBG-256 recorded an increase of 57.63%, 67.26% and 71.37% over NioxAvarodhi. BG-256 grown with Nio gave 68.97%, 75.38% and 78.00% higher values than Avarodhi under TPPW and 42.38%, 56.18% and 62.37% under GW application. BG-256 performed better than Avarodhi under different doses of nitrogen and with the highest dose (N30) root dry weight decreased proving toxic. Root dry weight showed increasing trend from vegetative to flowering and flowering to fruiting stage. 10. Nodule number plant* Like other parameters observed so far in nodule production also TPPW proved

87 Results effective in both the varieties. With BG-256 (Experiment III), TPPW showed an increase of 33.68%, 30.92% and 32.74%, whereas, with Avarodhi (Experiment IV), it gave an increase of 30.80%, 31.41% and 30.55% over GW at vegetative, flowering and fruiting stages respectively (Table 35). BG-256 gave better results than Avarodhi under TPPW as well as GW. 10kg N ha"* proved best among the nitrogen treatments tested in these trials and treatment proving more effective for BG-256 which recorded an increase of 67.24%, 78.01% and 72.56% over Avarodhi. BG-256 performed well under different nitrogen doses. Under^ N30 treatment, nodule number decreased in both the varieties, proving as excessive for these two cultivars of chickpea. It was also noted that NioxTPPWxBG-256 showed an increase of 68.76%, 77.93% and 74.00% over NioxTPPWxAvarodhi, whereas, NioxGWxBG-256 showed an increase of 65.21%, 78.11% and 70.74% over NioxGW^Avarodhi. Nodule number increased upto flowering stage and than it decreased. 11. Nodule fresh weight plant* TPPW proved beneficial. Therefore, wastewater with BG-256 (Experiment III) gave an increase of 21.15%, 21.38% and 25.59%, while with Avarodhi (Experiment IV), it showed an increase of 18.60%, 21.55% and 15.38% over ground water (Table 36). TPPWxBG-256 recorded an increase of 75.61%, 76.11% and 76.30% over TPPW^Avarodhi, whereas, GWxBG-256 registered an increase of 71.90%, 76.35% and 61.97% over GWxAvarodhi. Nio being optimum, thus, NioxBG-256 marked an increase of 69.66%, 76.37% and 67.52% over NIQXAvarodhi. It was also noted that the interaction of NioxTPPWxBG-256 increased the nodule fresh weight by 71.43%, 75.31% and 75.09% over NIQXTPPWXAvarodhi, whereas NioxGWxBG-256 showed an increase of 67.80%, 77.66% and 59.45% over N10xGWxAvarodhi. Like nodule number, this parameter also increased upto the flowering stage only. 12. Nodule dry weight plant' Wastewater of thermal power plant proved beneficial as it showed an increase of 26.42%, 28.71% and 35.23% with BG-256 (Experiment IQ) while, with Avarodhi (Experiment IV), it gave an increase of 21.13%, 36.00% and 25.00% over GW (Table 37). BG-256 responded better when compared with Avarodhi under TPPW as well as

88 Results

GW. It proved superior over Avarodhi as it gave comparatively more values under wastewater and nitrogen treatment. The treatment, Nio proved optimum for this parameter, being at par with N20 and BG-256 recorded an increase of 57.14%, 60.76% and 79.37% over Avarodhi under this treatment. The interaction of nitrogen, wastewater and variety Nio>

89 Results

the plant. 2. Total chlorophyll content plant"^ Total chlorophyll contents were enhanced by TPPW over GW in both varieties. The two varieties showed almost similar response with TPPW (Table 39). 10kg ha'' among the nitrogen treatments proved optimum for this parameter also, thus NioxBG-256 (Experiment III) gave an increase of 15.77%, 17.35% and 18.06% over Nio>

90 Results

41). Regarding nitrogen treatments, Nio and N20 proved equally effective being at par with each other and BG-256 showed an increase of 8.78%, 17.18% and 17.26% over Avarodhi because of Nio. It was noted that NioxTPPWxBG-256 recorded an increase of 4.23%, 17.34% and 12.30% over NioxTPPWxAvarodhi whereas NioxGWxBG-256 registered an increase of 14.28%, 17.56% and 22.82% over NioxGWxAvarodhi. For this parameter Avarodhi performed better than BG-256 under TPPW as well as GW and equally responded to nitrogen treatments. Leaf phosphorus content also decreased with increasing age of the plant. Potassium TPPW effectively increased the potassium content in the two varieties with BG-256 (Experiment III) it gave an increase of 15.02%, 16.98% and 10.24% whereas with Avarodhi (Experiment IV), it showed an increase of 15.27%), 14.06% and 13.37% over GW (Table 42). Nio treatment proved optimum, therefore, NioxBG-256 recorded an increase of 8.81%), 11.08% and 8.24% over NIQXAvarodhi. NioxTPPWxBG-256 registered an increase of 8.70%, 12.34% and 6.88% over NiflXTPPWxAvarodhi, whereas NioxGWxBG-256 showed an increase of 8.97%, 9.63% and 9.78% over Nio'^GWxAvarodhi. Leaf potassium also decreased from vegetative to fruiting stage. It may be noted that leaf potassium content showed higher values in general followed by nitrogen and phosphorus. 4.4.4 Yield and quality parameters The yield and quality attributes studied at harvest included 100 seed weight, pods plant'', seed yield plant'', biomass plant'', harvest index, seed protein content and seeds pod''. The significant data are described briefly (Tables 43-45). I. 100 seed weight TPPW performed better than GW in enhancing 100 seed weight in both varieties. With BG-256 (Experiment III), TPPW gave an increase of 3.14%) while with Avarodhi (Experiment IV), it registered an increase of 7.71%) (Table 43). For this parameter, Avarodhi performed better than BG-256 under TPPW. Among nitrogen treatments, Nio proved optimum BG-256 showed an increase of 24.28% over Avarodhi under this treatment. It was noted that Nio gave an increase of 8.63% over No for BG-256 whereas, for Avarodhi it showed an increase of 5.62%. Thus, it may

91 Results

be noted that BG-256 had boulder seeds than Avarodhi under nitrogen treatment. 2. Pods plant"^ TPPW effectively increased pod number in both experiments and with BG-256 (Experiment III), it gave an increase of 40.28%, whereas, with Avarodhi (Experiment IV), it showed an increase of 36.02% over GW (Table 43). It was noted that TPPWxBG-256 recorded an increase of 56.05% over TPPW^Avarodhi and GWxBG-256 registered an increase of 51.31% over GWxAvarodhi. Nitrogen treatment, Nio proved optimum and N20 was luxury while N30 proved deleterious and BG-256 showed an increase of 54.47% over Avarodhi under this treatment. NioxTPPWxBG-256 showed an increase of 56.03% over NioxTPPWxAvarodhi, whereas NioxGWxBG-256 recorded an increase of 52.33% over NIQXGWXAvarodhi. 3. Seed yield plant' TPPW performed better than GW in enhancing seed yield in the two varieties tested in Experiments III and IV, but increase was more pronounced in BG-256 (30.34%) whereas, it was 26.47% for Avarodhi. TPPWxBG-256 recorded an increase of 93.45% over TPPWxAvarodhi and GWxBG-256 registered an increase of 87.70% over GWxAvarodhi (Table 43). Nitrogen treatment of 10kg ha'' proved optimum being at par with N20 and NioxBG-256 recorded an increase of 92.12% over NIOX Avarodhi. On the other hand like some parameters N30 proved deleterious in both varieties. It was noted that for this parameter NioxTPPWxBG-256 showed an increase of 94.73% over NioxTPPWxAvarodhi, whereas, NioxGWxBG-256 gave an increase of 89% over NIQXGWXAvarodhi. With BG-256, Nio gave an increase of 31.83%) and with Avarodhi it showed an increase of 29.10%) over NQ. 4. Biomass plant'' Wastewater performed better than GW in increasing the biomass plant"' in both the varieties, but BG-256 (Experiment III) gave better values than Avarodhi (Experiment IV) as TPPW gave an increase of 20.03%) for BG-256 whereas it showed an increase of 16.70% over GW for Avarodhi (Table 44). Like other parameter, Nio proved optimum for this parameter also and its cmbination NioxBG-256 gave an increase of 58.19% over NioxAvarodhi. It was noted that NioxTPPWxBG-256 gave an increase of 60.61% over NioxTPPWxAvarodhi and NioxGWxBG-256 showed an

92 Results increase of 55.41% over NioxGWxAvarodhi. 5. Harvest Index TPPW effectively increased the harvest index in both Experiments (III & IV). Nitrogen treatments, were equally effective as the three treatments were at par in their effect for BG-256 (Experiment III), while it was at par with Nio only for Avarodhi (Experiment IV) and BG-256 gave an increase of 21.17% over Avarodhi due to 10kg N ha"'. It may be noted that Nio gave an increase of 6.04% for BG-256 while it showed an increase of 5.72% for Avarodhi (Table 44). 6. Seed protein content TPPW marginally increased seed protein content over GW in both varieties. Nitrogen treatment, Nio proved optimum for this parameter as it was at par with N20 and N30 and Nio>

93 Results

11.22% and 10.30% fresh weight over GW (Table 46). Treatment P30 proved optimum as P40 could not enhance this parameter, followed by P20 and Pio. An increase of 14.34%, 15.48% and 14.72% was obtained by P30 over control as TPPWXP30 proved optimum whereas lowest value was recorded by GWxPo at all the stages of growth. On one hand GW^Pao proved optimum being at par with GWXP40 and TPPWXP20 as compared to other GW-phosphorus combinations at fruiting stage. On the other hand, it was at par with TPPWxPio at vegetative and flowering stages followed by TPPWxPo, GWxPjo and GWxPio. For this parameter GWxPjo combination gave slightly less value than TPPWXP30. Thus it may be pointed out that P30 treatment can be used with both waters. Shoot length showed a linear increase from vegetative to fruiting stage. 2. Shoot fresh weight plant"' Application of TPPW proved efficacious over GW, giving an increase of 19.17%, 22.27% and 25.33% over control (Table 47). 30kg ha' phosphorus gave the optimum value followed by P20 and Pio, whereas P40 proved futile. Among various combinations, TPPWXP30 proved optimum being at par with TPPWXP40 at all the growth stages, followed by TPPWxPjo. GWXP30 proved optimum being at par with GWXP40 as compared to other GW combinations at flowering and fruiting stages but at vegetative stage TPPWxP,o was also at par with GWxPjo and GWXP40, followed by y TPPWxPo, GWxPio and GWxP,o. It may be pointed out that not only TPPWxPjo gave better results but even TPPWXP20 gave higher values than GW under higher phosphorus doses. Even without fertilizer TPPW performed better than GWxPjo, GWxP,o thus proving the superiority of TPPW over GW. A linear increase in shoot fresh weight was observed from vegetative to fruiting stage. 3. Shoot dry weight plant'* Between the two waters, TPPW performed better than GW, which gave an increase of 25.95%, 28.14% and 23.97% over control (Table 48). Phosphoius treatment of 30kg ha' proved optimum, being at par with P40, followed by P20 and Pio, whereas Po gave the minimum value at the three stages of growth. Regarding interaction effect, TPPWxPjo gave the optimum values at last two stages being a par with TPPWXP40, followed by TPPWxPjo. Among GW combinations, GWxPjo proved

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60 60 m •T3 OO r- 00 00 TT t-~-

C s > 3 r- r- •* r- ON VO -^ n m «n 60 > d d d

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> 60 •a .ti ON ON o CN (N CN 00 m 53 ^ VO n m CO >n vd CO CO 00 60 ^ fO 5 VO VO VO r~- r- VO I C •- > 3 « O s 0-7 _ 60

•^ o 0) ~ O CN »n 00 -•r ^ 00 0\ •^ r- S > ON cs o m •* 00 ON C3N VO 00° o" d in in ^ 1 o 3 O CI CO fO ^ 5- m m VO VO NO VO bo J tn >- 3 J: *-> s: Q. E II "C 3 o o o O o n o

JS t» a> Tf O CO CJN in o '•>5 cs »r> '^ •^ cd ON •"vT O 60 r- —; —^ TT «/^ a vd 00 ON •— ^ ON 4> o ^ cs 60 o I 60 o <— ^ — cs cs o o o > ^ 1 CO at) o a 2.S 0-1 b. H? 5 c o to ^ es o o CO )n «« CoO. i_ " ? O a> ~ • — I J3 c « O. CL, 1—1 to 23 eO c4 O to > .C — CO •^ n, C ^ c NO O —I CO 60 X?" IT) o O ^ > r~ 00 00 ov o ro CO m CO CO 'S t5 o € - 60 ^^ ON CS NO O "d- ON NO C3N O NO oo vo <^ 00 •4-* ^ c> ^ CO o «*- cs V 0\ c^• O ON CO NO 00 cs CO 00 60 ^ od o 2 ^ CO ro CO ^ •^ CO •§"^ b § CO o-r

CO

|o •S o OJ —' (^ ON ON r- <^ r^ CO CO o ^ -^ cs CO o ,-c \0 -^ o o ^ 00 00 r~- g 00 C3N ON oo' NO 00 o •^' ° 2 CO CO CO o ^ o cs cs CO •SI &-" CO >- «*-. S o s > CO *- -S a E w a 'c .a O ^

optimum being at par with GWXP40 and TPPWxP,o, followed by TPPWxPo, GWxPjo and GWxPio. It was noted that TPPWxPjo performed even better than GW with higher doses of phosphorus. Similarly, TPPW with 10kg ha"* phosphorus gave matching values as obtained under GWXP30 as well as GWXP40. Thus, it may be concluded that TPPW successfully reduced the use of fertilizer and can give better results even with low doses of phosphorus. Shoot dry weight plant'' showed an increasing trend from vegetative to fruiting stage. 4. Leaf number plant'^ Application of TPPW proved beneficial in enhancing leaf number at all stages (Table 49). P30 gave optimum values at all growth stages while P40 was wasteful and P20 and Pio proved deficient. Among various combinations, TPPWXP30 proved optimum, followed by TPPWXP20. Among GW combinations, GWXP30 gave optimum value at all the stages and it was also found to be at par with TPPWxP,o at vegetative and fruiting stages followed by TPPWXPQ, GWXP20 and GWxP,o. TPPW gave better results with all the treatments of phosphorus especially with P30, which also proved effective with GW. Even without fertilizer, TPPW performed better than GWXP20 and GWxPio. TPPWXP30 gave an increase of 40.04%, 42.19% and 40.21% at successive stages of sampling. Leaf number showed a linear increase from vegetative to fruiting stage. 5. Leaf area plant* TPPW also proved beneficial in enhancing the leaf area at all the growth stages studied (Table 50). P30 gave optimum value followed by P20 and Pio and P40 was not effective in comparison to P30. Thus, P30 recorded an increase of 33.49%, 34.36% and 34.49% over control and TPPWXP30 proved optimum being at par with TPPWXP40 followed by TPPWxPjo. Among GW combinations, GWxPjo proved optimum being at par with GWXP40 and TPPWxP,o at vegetative stage but at flowering stage it was at par with GWXP40 only and at fruiting stage GWXP40 was at par with GWXP30 and TPPWxP,o. It was noted that TPPW proved effective over GW when interacted with various doses of phosphorus, even TPPWxPjo showed better results as compared to GWXPJQ and GWXP40. A linear increase from vegetative to fruiting stage was observed in case of leaf area.

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c «,_ J5 o n 00^ ON o c r~ o eo ON ro •* 00 ON O 0) t^ ON (S U ^—'

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0 O ^—' 60 60 c n r- o ts 00 in C vo •—I 00 'C t^ ^ 00 o o ^ as o o •c »/n ON (N o o ^ u rt ON ts r-' r~-' Q PL, o o ^ (S cs O o o C 3 c 4> > ^ ON ** t^ ,_• fo JO o VO cd O o m ON u oo ••^ 60 E g o o 60 «/^ 00 ON ON ii :=: 11) o o -- 3 >

CO c 60 o 5 C o ^ c o o CnO. CO a, «> o o •«-• o J3 CO o. •o V T3 U o ^•^ ri 5 60 (rt tx ti. /—V m *-> c i£ CO c ^ a> i •£ r-c- — ON fO fO •«1- ON 00 60 vo VO — ON ON 00 t^ ON ON r^' fW vo ^' o oUi fO VO 00 NO vo -- iS 5 VO VO VO r- t^ C o a> G, O ^ ^ CO u o. b a> ^ o S^^ CI2 , -ca . J2 e« C4 > 60 60 I ° 00 r~ ro O 00 >n TT (S »r> —' u a> C x: > ^ r~ — — ON ON fS 00 — ro m d wS i— vd f<^ en 60 vo t^ t^ 00 00 2 c c cs r«^ m >iO vn > 3 « ?^ eo- r«

^ o ON 00 ^ ON O r— »n (N rr CO ro ro vo t^ 00 vo VO r~ r~ ON Tf t-~ o o c o ON f—I CO o o m ro § 60 vo vo r^ r- O ^ -^ •« to CO w- 60 Cn « (U ^ -C

^« S.E ^ >• 'C 3 W o o o o o OH o ^ cs f^ r^ a. a. o CU OH CU OH PH II o o o o o o 3 "* Z Z o C/2 O to S Xi z Z z z z z z « £<<<

6. Branch number planf TPPW again proved beneficial as it gave an increase of 22.07%, 23.22% and 21.17% over control (Table 51). P30 gave the optimum value followed by P20 and Pio whereas Po gave the lowest value, at all sampling stages. TPPWxPo proved optimum at all sampling stages followed by TPPWXP20. Among GW combinations, GWXP40 gave maximum value being at par with GWXP30 and TPPWxPio at vegetative and flowering stages, while GWXP30 gave similar results as that of TPPWxPio at fruiting stage followed by TPPW^Pao, GW^Pzo and GWxP,o. P30 proved equally effective when interacted with both waters but TPPW-phosphorus combinations were comparatively better than GW-phosphorus combinations. Branch number showed linear increase with the increase in growth. 7. Root length plant"' TPPW gave better results than GW. An increase of 18.96%, 27.59% and 26.82% was registered by TPPW over GW (Table 52). Phosphorus treatment of 30kg ha"' proved optimum followed by P20 and Pio. Regarding interaction effect, TPPWXP30 proved at par with TPPWXP40, followed by TPPWXP20 and GWxPo gave the lowest value at all the sampling stages. GWxPjo proved optimum as compared to other GW combinations being at par with GWXP40 at flowering and fruiting stages. On the other hand, it was also at par with TPPWxPjo at vegetative stage, followed by TPPWxPo, GWxPjo and GWxP,o. Treatment P30 proved effective in enhancing root length when interacted with both the waters. TPPW phosphorus combinations proved beneficial for this parameter over GW-phosphorus combinations. Root length showed a linear increase upto fruiting stage. 8. Root fresh weight plant"' TPPW successfully increased the root fresh weight and thus proved beneficial for this parameter also (Table 53). Treatment P30 gave the optimum value being at par with P40, followed by P20 and Po, whereas Po gave the minimum value. TPPWxPjo gave the optimum value followed by TPPWxPzo and GWxPo registered significantly minimum value. At vegetative and fruiting stages, GWXP40 gave the maximum values as compared to other GW combinations being at par with GWXP30 and TPPWxPo,, while at flowering stage, TPPWxp,o gave the maximum value being at par with

96 c 5 00 (N o ON o 60 c W-) •^

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GWXP40 and GWXP30, when compared with other GW combinations, followed by TPPWxPo, GWXP20 and GWxPio. TPPW gave better results with all phosphorus doses at ail the sampling stages, whereas P30 proved effective for both waters. Even TPPW without fertilizer proved effective over GWXP20, GWxP,o. Root fresh weight also increased linearly. 9. Root dry weight plant'* TPPW also proved effective for dry matter accumulation at all the stages of growth. It showed an increase of 26.32%, 33.18% and 24.85% over GW (Table 54). Treatment P30 as noted in other parameters, proved optimum. P40 proved ineffective in comparison with P30 while Pio and P20 were deficient. TPPWXP30 proved most effective combination as it equalled the TPPWXP40 followed by TPPWXP20 and TPPWxPio at flowering stages, whereas at fruiting stage TPPWxP,o was at par with

GWXP30 and GWXP40. All the TPPWxphosphorus combinations except TPPWXPQ gave comparatively better results than GW-phosphorus combinations at flowering and fruiting stages, while TPPWxP,o gave similar results as obtained under GWXP30 as well as GWXP40. Root dry weight increased linearly from vegetative to fruiting stage. 10. Nodule number plant' TPPW also proved beneficial in enhancing the nodules at successive stages of sampling. An increase of 21.19%, 22.68% and 20.30% was recorded under TPPW over GW (Table 55). Like other parameters, P30 gave the optimum value at all the growth stages. It was followed by P20 and Pio, whereas P40 gave statistically equal value as obtained under P30. This treatment showed an increase of 27.28%, 28.12% and 26.24%. Among various interactions, TPPWXP30 proved optimum. Among GW combinations, GWXP30 also gave the optimum value. It was noted that TPPW proved good while interacting with various doses of phosphorus over GW-phosphorus combinations. Even TPPWXP20 showed better results than GW combinations with higher phosphorus doses. TPPW without fertilizer also gave better results than GWxPjo and GWxP,o. Thus, it may be concluded that by the application of TPPW we can easily save the inorganic fertilizer. Nodule number showed an increase from vegetative to flowering stage then it started decreasing at fruiting stage.

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o (L) o O

(30 60 •a c t s VD vn m o ts ts

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2 60 5 .E C af> J3 o(> rt 0.13 u c/al. k. O u J3 c S "H- O^ z «« c ^ O ON «r> en 00 iri VO O OO en 60^ 00 O C^ t~- 00 O « O VO CM O O o c^ (N >o >in m en '^r VO VO CS CS (S oi CN •s t> •«^ ^ (U

rt je J5 > ^^ t^ CNi »n r^ ^ VO g) CN VO 00 cvj r>- ON t^ en m TT en NO iri 00 00 >N M m •^ «/^ r~ r~ CN en •^ t-- r~- s° 60 5 CS CS CS CS CN «t- CS > O

o S ^ •c .2 o o o o O CO O — O. 2 o d d d d s d d d c o «+H IS o (/> o T3 £50 60 0^ 60 ON >n ^H o t^ ^ m ^^ 0 en ON CO ^ vo — m o •-< o ^ ts X5 Tj- m m r~ t~-

CO cs >n m Tj- 00 m *>° f*^ ,^ vo o ro o o '5CO *^ 60 fo rr rr w-i v^_ •4-* o — c/3 60 g u60 o to d d d d d d 9 o. Z o « > o o §1

^ 60 2 60 2 .= a H 0 0 •4-* J3 0 K" <« CU CO »J l-r g 5 0 V •a ^ «^ I J3 4-» •<-' _ c « & "J O, V CO CO en 't tA "* US •^ CO c ^ to o m »n >— o "O c rl- (S O rl- O 60^ ON •—' ro f- t~~ CO O t~ CN •^ iri O U — CS CS (S (S NO v£) r- 00 00 i= .> « > d d ci d d d d d d d « UH k. ES

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3 x: > CO 0000 O — CN m fl 4-* o — UH UH U' Pu UH PL, tlH UH b UH s Results

11. Nodule fresh weight plant" Regarding nodule fresh weight, TPPW proved good at all the stages of growth. It registered an increase of 21.18%, 19.37% and 21.09% over control (Table 56). P30 at all growth stages proved optimum followed by P20 and Pio. TPPWxPso proved optimum being at par with TPPWXP40 followed by TPPWXP20. Among GW combinations, GWXP30 gave the optimum value being at par with GWXP40 and TPPWxPio, followed by TPPWxPo, GWxPjo and GWxP,o. TPPW proved beneficial for nodule fresh weight with phosphorus doses. Thus, TPPWXP20 proved more effective over GW-phosphorus combinations (P30 and P40). TPPWXP30 showed an increase of 55.12%, 53.51% and 55.72% over control. Nodule fresh weight increased from vegetative to flowering stage but decreased from flowering to fruiting stage. 12. Nodule dry weight plant"^ This parameter was also affected significantly under the waste water at the three successive stages of growth studied in this experiment. Therefore, TPPW gave an increase of 23.42%, 23.96% and 20.65% over GW (Table 57). Phosphorus doses at the rate of 30kg ha"' proved good followed by P20, which was found to be equal in its effect as that of Pio at vegetative and fruiting stages. Among the various interactions, TPPWXP30 proved optimum being at par with TPPWXP40 followed by TPPWxPjo. Among GW combinations, GWXP30 proved optimum. TPPW as well as GW performed better when interacted with P30. It was noted that TPPW gave better results even with Pio, as it was at par with GWXP40 and GWXP30 confirming the earlier observations where TPPW proved beneficial in terms of phosphorus availability. Nodule dry weight increased from vegetative to flowering stage only and it decreased thereafter. 4.5.2 Physiological parameters Physiological parameters viz. leaf nitrate reductase activity, total chlorophyll contents and NPK were studied at vegetative, flowering and fruiting stages of growth. The significant data are briefly described in the following pages (Tables 58-62). 1. Leaf nitrate reductase activity (NRA) TPPW also increased the enzymatic activity thus it proved better than GW. It gave an increase of 14.93%, 14.15% and 14.01% over control (Table 58). P30 proved

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> (0 _^ 2 2 60 2 .S C 2 0 a, o CO c3 0 JS 0 x: «5 0. c4 C/1 i-i I 2 0 V -rl 4> I J3 5 "Q. ^ a. OH *- O C f ^-^ « ca o. c ^ "* o VO O NO "<1- ON V~l 60^ 00 o m t>- in NO O V ^ > ON r-' ON cs ON ON i3 .> «0 «o NO NO •^ •* »r> m IT) 9, « ^ a. 0- — E fc•-> u ^ :S^ 0) 60 "^ ^ > CS "«1- ON 00 t^ 00 *rt •* 0 r- NO cs c -c CS «0 CS V~t VO cd > r~ 00 m r>- cs m IT o T^ r~ t—I 0\ ON rn iri »r> 00' 60 «n VT) NO vO NO cs 2 m_^ u-> K NO NO Vt NO > 3 « s '^ to -^ i« *^ ^ o JC -^ ffl •^ o •o O vr> >— fO ON cs •^ 0 TT U-) ro fO •/^ O — —I NO 0 0 0 f- 00 t- r- ~ t- m vn 00 ro r*^ 00 1*3 1 ^^^^ Ti" W-) »/^ W-1 I •* •^ ir> »n •^ 60 J •^ -a ^ U U 3 x: > CO K E o o o o 0 0 0 0 •c 3 o — C4 f*% r^ 0 0-1 OH A4 &< OH Oi Oi OH O<*H^ OH X) 00000 0 0 0 0 0 3 Z Z Z 2 Z Z Z c g 00000 0 0 z0 z0 z0 tS CQ cd < < < < < < < < < < UH b PL, b UH b UH UH b i^ Z u 00 C c •—' o o o o ^ 00 00 00 o o o o o (A o o o• o ^ —^ 000 c '^ o J*o- C/3

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t*-c CO o ^—.^ M 60 c>^ 60 O ^"^ C! 00 ON O (S c •"^ l^ ON J* in u tS ^ VO O O 'C o o o •^-4- * V o o o c ^ g^*: CS •O « r~ ^ fs ^ m 00 O »-' •^ •^ O — C/3 — (S tS S^ __^o b5x 52 60

(X, u H.S c fr ^ o vH^ **J eS o Ui ^^ 0) c/i (A rt 5 u ^ o o O JS CI-T3 •« o« a* »5 «2 K^ 5 o ^ o. r-N ed S't; V—• CO c3 o c 00 o VO t~- m ON (S m C ^ CO o ^ 00 o\ ON f—t 1—4 "H-iS 9i •—1 60^ 1- 4^ o o o •—' O 0) S O o O O o o o o o o i= .> ^S •£ t5 O ti -^ ex _• u B3 .U X! 'cd en .c 4} ^ a 60 ^^ '5 •^ ro O •^ 00 ro r- m IT) IT) en ON o o (N (N| 1—t C ^ P O ^^ •—' •-^ •—* »—» cd C 60 1^ o o O o o O "2 O o o O o O > b ^1 o- ^«-^ VO u. "^ cd U CS «-> 1 *-• o 00 VO - ON in o r^ (S O —' Ov O o t^ 00 ON O O ON O O o o — o g o 1 O O o o o o O O '~v o o o o o •o td 60 1-1 CO ^ (L> U 3 j=: Jd ^ *- s: > cd K E W S o o o o o o o o •C 3 •*^ n m o n n n o CO C CU OH Ox OH c/1 e« r-' o o a. (X ftx cd o o o o o o o .o vn o 3 C/3 O •«-s» Z 2 2 o o Z c c

optimum P40 luxury while P20 and Pio deficient at all the sampling stages. TPPWXP40 was at par with TPPWxPso followed by TPPWXP20. Among GW combinations, GWXP30 proved optimum being at par with GWXP40 and TPPW^Pio followed by TPPWxPo, GWXP20 and GWxP,o. The treatment P30 gave better results when interacted with TPPW as well as GW, whereas TPPW performed better than GW with various doses of phosphorus, even TPPW without fertilizer gave higher values than GWxPjo, GWxPio indicating the superiority of waste water over ground water. Enzyme activity decreased from vegetative to fruiting stage. 2. Total chlorophyll content Total chlorophyll content increased under waste water application at all the stages of sampling. Thus, TPPW recorded an increase of 9.47%, 12.00% and 11.69% over GW (Table 59). Phosphorus dose of 30kg ha'' proved optimum followed by P20 and Pio. TPPWxPjo gave the optimum value being at par with TPPWXP40. GWxPjo performed well as compared to other GW combinations as well as TPPW combinations except TPPWXP30 and TPPWXP40, which gave the maximum values.

Treatment GWXP20 was at par with TPPWXPQ at all stages of growth, which proved the superiority of TPPW. Total chlorophyll content decreased from vegetative to fruiting stage. 3. Leaf N, P and K contents Nitrogen TPPW enhanced leaf nitrogen content when compared to ground water. The increase was 15.49%, 15.20% and 14.98% by TPPW over control at successive stages of sampling (Table 60). Phosphorus treatment of 30kg ha'' gave the optimum value followed by P20. Among the various combinations TPPWXP30 proved the optimum being at par with TPPWxP4„. These were closely followed by TPPWXP20 at all the stages of growth. TPPW proved good when interacted with phosphorus thus, TPPWXP20 gave better results than GWxPjo and GWXP40. Leaf nitrogen content showed a linear decrease from vegetative to fruiting stage. Phosphorus Regarding leaf phosphorus content, TPPW proved superior over GW. It registered an increase of 16.97%, 16.06% and 15.03% over control (Table 61) P30

99 •a JO c c« 60 C ON 00 00 CN _C ON r- CN —' to (U to O •ll 60 60 •a C tS ON OO VO VO "3 VO CNl O —; ON m OO to OS vd ts e« o OO 00 00 X) CO Ui o cd .S 00 00 00 Q U 00 00 C-- ON 00 ti "^ ON o 00 U~l O m o ON VO •^ CD <4-l _aj o CO 2 60 a- g E C CO H C a. o o tn CO o x: CO CO 0^ to 4-^ O u c ^ OH C^ CC 1^ CO ^ s "—' c« *^ 60 00 »n »n 1^ C C O 00 >r» O -=!• (S o cues cd ON t~~ r^ ON in m ON r- «^ (S ON ON

a> <" to 13 '-> E J

J3 a •^ s: > 60 O m O 00 in r-- o 00 •^ c in •<1- VO (N

^^ - o Ov »ri CNl ON ON CN| ON CNJ VO 4» _ (^ 00 rr •—; CN (S '— •^ ts •^ *r) o •« 3 JC 13 •"

o o o o o o o ts r> f*^ G PL, OH CL, OH OH 00 o a< OH OH U o o o o o o o 3 *-' Z ^ Z 2 Z C/3 O 6 o o o 2 C •4-* o o o o o z CO < < o o r- m m *••-» ^ CN f^ 00 '53 o\ o o m r<^ o o o o ^' cs tsi ri (S 5 o o o c (U o « o 00 •a 00 00 c m cs 00 r~ r~ 00 ON 1/3 60 ^ cN m •J- CS 00 vo t^ c o o o X) —• ts cs (S (S Q o o o S I O 0) 00 (It o .2 o^ t"S" '5 3 II (U O. 00 ^ ^ ^ o\ > r- r- 00 S «u o 00 r- ON ON — CS fO 00 00 ON •—1 '— u 00 ^ ^ ^ (S C^l 00 o o o ^j CO > o o o

2 5 00 B..c Q ; ^•^ c ST =« ^1 o u ^^ a O. T3 ^>-^ CO -a .Si « > u^ ^c OH I—I

%^

CO J= 00 vo o ON vo vo (T) VO VO r/^ '— -— CM O D o >—1 Cv| VO t~ C7N ON •- > CS CS fS tS o I* CO O o

> •^ 00 in 00 •^ •s ^ ON CS OO O 00 TT r- »r) Tt vo ^ m fo W-) •^ o CM ro m ^ ts t^ vo vq VO t~- 00 o O 00 <. - rO^ S > 00 CS (N

> 3 "* SI O- o •=: i-s <-< >«» cd ^ •^ o f^ ON ^^ t-» m r~~ «r> IT) T}- O 1 ^ 00 vo m 00 ON Ov ON vo CS O —I o ^ ON O •—' m f<^ rr «n vo 00 00 c o % •— ri (S CS CS SQ -° 13 CO '-' OOw V Hi «*- eO 3 X O V •- O- tj ^ S O

W o o o o o o •C 2 o o ^ «N n m 5^5 C5O^ Ov OH Ol P^ OH 0-( 0^ OH OH 0-( o ^ ci m o 3 o o o o o 00 o z z ^ 2 z ex z ;z 2 ;£ z o o CQ o o o o o < < < < < PL, b pH tL, b 2 <

TPPWxPo, GWXP20 and GWXPJQ. It may also be pointed out that even with higher doses of phosphorus, GW could not show better results than TPPW. Phosphorus content also showed decreasing trend from vegetative to fruiting stage. Potassium Application of TPPW proved superior over GW as it gave an increase of 14.69%, 8.83% and 14.75% over control (Table 62). 30kg ha"' proved best among the phosphorus treatments followed by P20 and Pio Combination TPPWxPjo gave the optimum value at all sampling stages. TPPWXPJQ gave an increase of 38.70%, 30.60% and 36.84% over control at vegetative, flowering and fruiting stages respectively. Leaf potassium content also showed decreasing trend from vegetative to fruiting stage. It may be pointed out that potassium content were maximum among the three macronutrients followed by nitrogen and phosphorus. 4.5.4 Yield and quality parameters The yield attributes included 100 seed weight, pods plant"', seed yield plant"', biomass plant"', harvest index and seed protein content were studied at harvest. The data for yield characteristics were mostly found to be significant and are described as follows (Tables 63-65). 1. 100 seed weight In this case the effect of waste water was found to be non-significant. Treatment P30 gave the optimum value, being at par with P40, followed by P20, which was found to be at par with Pio, whereas Pio was at par with Po as noted in seed yield. Treatment P30 gave an increase of 7.43% over Po (Table 63). 2. Pods plant"* TPPW proved effective in increasing the pod number in comparison to GW as

100 -a c CO on c 00 •<3- en o o\

u o o •o •^ 60 M "cO 00 •^ VO r~- <^ VO m c TJ- i ro (*i Q o E o O o '5 c 3 DO O > eO m rs I—1 CO >r^ rf fo C7\ r^ 1—1 O o •^ kO TT CO TT VO ON •«1- \r) VO oo 00 VO V 60 CS (N CS (S CS CS 60 o o o > o o o

> 60 §: « 2 E H != 2 C « <« 4-* n o l-i -«-> j= () U CO Q. CO o 1?! en k. eO B5 O u i J3 -o .2^ 11 OH s to JS CO g .^ I/) > o ^~^ CO <-. ^ fe a. c ^ r-H c •^ (N 00 60^ 00 cs (S o r~ O U c/i r~ VO »r> ON Ov uCO m o VO CN CN ^ > 0) V VO 00 ON rn <^ •^ kD »ri r- r- fo en to ^ •^ 1- o

rs ^

^ -a 0) *- j= 60 > VO Tl- ON VO (N C K^^ VO Tj- r- ov ^ fo 00 fn m ""l- •r~^- ^ M- O VO -"fl- ON o (S r-; r- r<^ iri VO VO 00 00 60 ^ en '^ •^ '^ "^ •^ 2 > ^ I 3 « o feb o-r ill ''^ c to •" o ,—1 t) — •o o O • ^ •^ •^ VO VO O o r-i •"a- •^ »o VO go g ro r<^ r»i Tf Tj- m s^ o -o 1o 00 J ° a 3 J= •^ s: > CO

Ui h •C 3 o •-^ cs f^ n •4-* J^ •« CL, CU CL, CL, CL, CL, CL, o a O. 0^ &< o o o S3 t« < < < < < a^ U < <: < < < OQ 1 H tL, H U^ u-< b b U- :s U-( tL, UH U-, 1^ r^ r- O B (S CO •^ ed 00 ON O CN m o o o c3 (U t-^ • ^M 60 ts cs CN o o o O :s o O d d d 2 d d d Cu B ^o J3 O t*- (L> o M o ^—.^ 60 I-a "cd o\ r- •^ 00 •^ B (N •* in C/3 4_t 'C ^^ o O •

,—1 > _§ w »o 00 ON ts m '•^ ts -^ in •5 a> r- 00 00 o r-H ON cd o o o 4-» o< 00 »-H f—1 (S ts T—« o o o 1 o o d d d d 60 d d d •Sijs > §1 ?^ 2 I PH U OH >= B -§.&

H^•^- ^ -. 5. 2 _o 4-* o u « JS o U CO CU cd 4-J Vi u 5 2 U ^ o Cd o U f cu *-* «!: to O. B 1 S *- o. B ^ n «4-i ^ o c rr o >r> r^ VO B ON VO o r~ o 60^ "3. «'5 CO t^ ON o *n cd TT m vo r~ O U 0> >/^ 00 1^ "cS CO <^ •^ •^ TT fe > o O d d d o o o o o o ^iJ O *i o. c O) u T; 1-" ife RJ ,4}

sUi e•-" V T3 a> cd "^ ;>. ** •£ J= _> 00 . ""^ J^ •"^ •«-> <^j J.^^^ cs o\ VO ON CO 00 _c K^ 00 VO — O r^ VO C3 ^^ o <^ 00 Ov TT ^^ in VO r^ ON ON ^ ^t rf •^ •^ •«r ^ CO c 60 E> o O? d d d d 1 I> b o o o o o > 3 «^ ^1 o-r ^—'vO 0o0 I- «/^ cd 4> CS 60 4-* 1 •^ o

VO o •^ VO Ov r^ ON in ON • fo m CO •^ •^ CO 1 = g o o g d d d d o o o o o l-o td eio>-] • CO L- 3 J: 4- R tJ ^ > cd iSi ^ o, B U ^ o o o o o o o o ''^ 2 4-( o cs r^ (*i •^•t B OH OH CL, CL, pL, B o ^ C4 r^ f*^ 1 SS "5 ^•^ U o o o o O U OH 0-, CL, CU OH 6 E o o o o o •2 «« o z z o •4-* B Id CO Cd cd 0) o o 0) z z z z z CO < < < < < o o o o o H H b UH b b b H Z < < < < < UH UH (JL, tL, UH T3 C tso C ^ ON 00 r- VO VO VO IT) V c TJ- •^ rj- OO 00 c ^ (N ro 60 vo r^ 00 r-. ^ O 2 (S oi r4 rW rW o o o o o o c o o (30 00 13 VO •—' ON ''^ •—' •5}- 00 ON O 'C fn vo vO (N ri *c --I C^l -"T to r~ 00 ON m r^ o o o o fS (N (S m m Q o o o o O E 5 3

> VO VO t- VO ^^ t^ ON O (N •^ (S TT m cn o\ 4-* ^ ro 5 >/^ VO r~ o O r- OOP DO CNJ CN (S f^ f<1 fNj 00 o o o >

2.:c0 0 c o o P. o en o. -a O (U J3

c ^

ON VO r—1 o VO '^ TT r- ^ ON 00^ m m o fN CNI >o rj- fsj r~ r- p oj •—' m in O o -^ fs f - u O D.

S -= > 00 VO (S r^ o 0 Tt •^ Tj- V-) »r^ a ri fs ri (N r>i > eg O- S « CO -*-

*- o VO — rr m ON •^ r~ ON VO rr r> Tf ON VO VO — ON »n 00 ON 00 O — VO VO c> o —mm m Tj- •^ 'sT CN) (S (N -

> CO

•C ,2 o — fs r^ n _ o ° o c« Cb PH OM CL, &4 O — fS JO £0 o o o o o CU CU AH 3 *-» ;£ ^ z 2 2 2o Zo Zo Z Z CO O o o o o o PQ o ^ o o o < < < < < < < < < < b U^ pLi UH b Z UH U-c (IH UH til Results

it showed an increase of 25.49% pods (Table 63). Like other parameters, P30 gave the optimum value as P40 proved futile. It was followed by P20 and Pio, whereas the least value was recorded by Po being 32.20% lower than P30. Among various combinations, TPPWxPao proved best as it was at par with TPPWXP40. It was followed by TPPWXP20 and TPPWxPio. It is to be noted that not only TPPWxPjo gave better results but even TPPWXP20 performed better than GW-phosphorus interactions under higher levels, followed by TPPWxP,o. TPPWxPao gave an increase of 65.50% over control (GWxPo). 3. Seed yield plant* Application of TPPW proved beneficial in enhancing the seed yield over GW and the former recorded an increase of 21.44% over latter (Table 63). Among the various doses of phosphorus, P30 proved optimum, being at par with P40 followed by P20 and Pio and expectedly the lowest value was registered by Po. Treatment P30 recorded an increase of 27.86% over Po. Regarding interaction effect, TPPWXP30 proved optimum, being at par with TPPWXP40, followed by TPPWxPjo and the

lowest value was given by GWXPQ. Treatment P30 gave optimum values when interacted with TPPW as well as GW, but with TPPW it gave better results. It was also noted that TPPW without fertilizer dose gave better values than GWxP,o and even with GWXP20 indicating the utility of waste water as a source of this important essential macronutrient. Similarly, TPPWxP,o was at par with GWXP30 confirming the above observations. It may be pointed out that among the various levels of phosphorus Pio and P20 proved deficient while P30 optimum. On the other hand P40 was at its luxury consumption as it could not enhance the yield further in comparison to P30. 4. Biomass plant'' TPPW performed better than GW. It showed an increase of 20.31% (Table 64). Among the phosphorus treatments, P30 proved optimum being at par with P40, followed by P20 and Pio. Among various interactions TPPWXP30 gave the optimum value being at par with TPPWXP40 followed by TPPWxPjo. It was noted that TPPWxPio gave better results than GWXP20 and GWxP,o, and TPPWxp,o was found to be at par with GWXP40 and GWxPjo whereas GWXP40 was found equally effective

101 60 c

2 C U m VO O >n in n Ov m u TT rf in »r> vO '>% f-H r- m 60 >—1 r—» (N O r-^ TT Vd —J ^' f*^ m m •^ •^ o o o CO in c en -Oo a CS CS rf O O 00 en 00 CS VO BJ Vi O ov r- X) ^ 00 o vd vd O 'it Q m t-~ -" H U o o ^ 60 3 J3 60 to Si I 2 '53 ? O ov vn ^- 00 V ^ 2^ ^ o 00 00 60 4> O 00 vo en r^ vd ov "^ A to O &, o fs vd z O z O q_i CO fO m ^ O C "" o "33 5 .S c |i o "Is OH 0) o Q. O.'O u ^ c

cd C ^1 C ^ >n VO 60^ c vri in Ov ^ Ov VO O V t-~ OV O oo' oo' ON O o is .> r- 00 00 o- 2 •c t5 /-> O, o CQ —, > J5 in .c .60 '53 •a ^^ ^ 5 00 r^ Ov o m o 0) «n o vq 00 00 00 •>> o o l m m v ov •^> 00 § KJ ? ^ o »^ S ^ o?* •.-> o fO M- OV 00 t-- m VO >—I •*-• t*H

> «j

'C .s3 W "H. o o o o o "Vi o o — r« m fi CoO CO o Cu Cu Cu OH OH ^ «J VO 3 4-» o o o o o 00 O — z z z z z a < z z z z z OQ H o o o o o < < < < < z UH UH UH bu tu Results

with GWXP30 and TPPWxPo. Thus, it may be pointed out that TPPW with Pio or even without phosphorus dose gave matching values as obtained by GW with phosphorus. So, TPPW can successfully reduce the use of inorganic phosphatic fertilizer. TPPWxPao showed an increase of 46.60% over control. 5. Harvest Index Effect of waste water was non-significant. Phosphorus treatment P30 proved best being at par with P40, followed by P20, which was at par with Pio and this treatment was found to be statistically equal with Po. The optimum treatment showed an increase of 4.67% in harvest index over control (Table 64). 6. Seed protein contents Protein was not increased under TPPW as observed in various parameters including seed yield. However, it must be admitted that it was not decreased either, as the effect of waste water was non-significant on this parameter. Therefore, indirectly seed protein was enhanced under TPPW as it increased the seed yield plant"'. Among various phosphorus doses, P30 proved optimum, being at par with P40, whereas values given by Po, Pio and P20 were at par indicating lesser role of phosphorus. P30 recorded an increase of 5.42% over Po also confirmed this observation (Table 65).

102 a u C o C 00 00 •^ ON (N o 00 0 0 00 C/D 60 00 ON ON O ^1- •<4- z 5 z

til o •a •o .S •4-J •a "S CO 00 00 in o —I CO a c o la ^ Tj- 00 ccS 00 as OS (N f»1 •^ X) t d o o o

CO c2 X °c 3

a o. t<3 60 00 c N—' ON 00 Os o 60 M VO t^ r^ 00 00 CO % 'it •* Tt Tf TT cd 4 - E o. tn o CO Ui Si O « c CO 00 o 5.S CO ? 5 OH 0 OH J3 0. H CO C1.T3 5.^ 0 J5 T3 • — *i 0. «i a. 0 6J CO 2 CO -4-* to > C ^

•4-1 0 00 ON 60^ •4-c* VO O "S. I-I S^ 0) h2 /-> G, ^ •5 <* to

o .•4-c* o< J3 CC •3 '% CO H £ c 0 ? VO 4= S 0 t^ ON CN ^ O 4-» «*; CN TJ 60 00' 00' ON 0 C 0 CN °-o n) «r) CS § rt 0? bffl v—^ I- >• 0 «^ 4-u* •^ O CQ ^~\ iri ^ J 0 •^ fn 00 •0 C 5 IT) iri vd 00' 00° vd § 2 3 0 .5 U-i *>.* CO •- 60 C4-1 b 0 «3 •4-* »» U ?> .4) 0 •s-i 0 0 0 0 0 0 n n O to OH cu CO CO •«<•• 0 0 0 X) ctj vO 0 D O (1> 0 Z 0 Z c Q. 0 2 0 a ^ CQ cd < < < < H < Pu n3 •T3 a C ca O c IoS 00 en 00 00 4; O o 00 O. z z z c -a o o a C/3 CO O (U -a T3 u O OS CO to s (/) J2 00 •o cu C •o •0 s0 ^ s ^ Q V 0 B u C/3 U OD a c •«-» 'c c 3 o o c CO o o o 00 o. •a u u o c o 2 00 B .S O CO

^ 2 CO "" H a, 1- O Eo o, -a Vi -~ cd V i >

•*-» "* VO >n O C *i VO c ^ cd C •7Q; . u« 00^ ^ t2i O U « C t! .> c v u *- O 4) ^ O •o s c 5 TT ^ ^r >n »n D S 1 -a t3 o 5: CO <- 60 .Si «*- fc 3 J= O Q *- v. u CO ,ti u UJ ii' 0000 •C 3 o — r^ f*i « J^ '^ iri OLH CU CUi CU OH 52 CO VO o o o o o "S «» 3 •*-• Z 2 £ Z 2 00 o O O O O o PQ < < < < < UH tL, Ui tU b discussion Chapter-5 DISCUSSION

A perusal of the results described in the last chapter clearly indicates that irrespective of all other inputs, chickpea performed better when irrigated at regular intervals with wastewater than with groundwater. Obviously, it could be due to optimum growth and development of this crop under wastewater. Let us, therefore, consider the presence of three major essential macronutrients, viz. N, P and K, in addition to S, Ca, Mg and CI (Table 5) could have played an important role in this ameliorating effect. Let us now consider the roles played by each of these essential nutrients one by one. N, for example, is the single most important element limiting plant growth and is invariably required in large quantities deserves special consideration in this regard. As vegetative growth includes the formation of new leaves, stems and roots, the involvement of N through protein metabolism controls the growth. This is clearly indicated by the observed optimum growth (Tables 26 to 37; Figs. 10-13) in the present study. It may also be added that, on application to soil, most of the non­ organic forms of N remain readily available for uptake, during vegetative plant growth. In comparison, only about 5-75% of the organic forms is commonly mineralized and that too in about one year after application (Sommers and Giordano, 1984). This lends support to the above observation of the suitability of wastewater as a good source of this nutrient. Another aspect that requires consideration here is the fact that both NH4"^-N and NOs'-N were present in wastewater, the former being about five times more than the latter (Table 5). It is noteworthy that applied NH4*-N is toxic for some higher plants, including bean and pea (Maynard and Barker, 1969). However, in the presence of NOs'-N, it has been reported to benefit sunflower (Weisman, 1964) and wheat (Cox and Reisenaver, 1973). The observed nutritional superiority of wastewater (containing both NRt'^-N and NOs'-N) for growth of chickpea in our study is thus not exceptional. Nonetheless, it may be claimed as the first such report for a leguminous crop. Similarly, the presence of additional P in wastewater might have primarily Discussion

influenced root growth (Tables 12-14, 32-34 & 52-54; Figs. 5 & 12). It is known that for the effective use of P, various factors operate together, such as rooting pattern, length of crop growth, soil characteristics including pH as well as dose and source of P, in addition to the presence of water. Since wastewater was one source of irrigation and was comparatively richer than the other source (groundwater) by about 58% in all experiments (Table 5) the observation of improved performance of the crop (Experiments I-V) under wastewater is understandable. It is all the more noteworthy because application of phosphate fertilizers was its limitation as P fertilizer applied to the soil are very rapidly changed to less soluble forms and, therefore, become less and less available with time (Russel, 1950). Admittedly in short season crops, like some vegetables, growth responses to applied P may persist upto harvest. However, long season crops, like corn and chickpea, show only early growth responses and comparatively much lesser effect at seed formation and maturity. Frequent wastewater application until this late stage, therefore, enhanced P availability to the crop and ultimately le?id to higher seed productivity (Tables 23, 43 «& 63; Figs. 9b & 16b) in chickpea. Let us now consider the effect of the third major nutrient (K). It is well known that N is fully utilized for crop production only when K is adequate (Mengel and Kirkby, 1982). The presence of K in almost double the amount in wastewater than in groundwater (Table 5). Therefore, ibenefited the treated crop not only due to its own physiological role (Wolfe/ al., 1976) but also by enhancing the effect of N. While it increased the chlorophyll content of alfalfa leaves and also the CO2 exchange rate on plant'* basis (Collin and Duke, 1981), it is not surprising that this nutrient (along with Mg) improved the chlorophyll content in the present study also (Table 19, 39 & 59; Figs. 7b & 14b). The presence of higher NPK contents in leaves (Tables 21-23, 41-43 & 61-63; Figs. 8 & 15) grown under wastewater further confirm these observations. This ultimately led to increase 100 seed weight (Table 23, 43 & 63; Figs. 9a & 16a) and seed yield (Table 66). In addition to N, P and K, presence of S also improves growth and N fixation (Walker and Adams, 1958). Therefore, in our study S as well as Ca and CI present in wastewater (Table 5) might have contributed further towards

104 Discussion

enhanced growth and led to the promotion of the crop's yield. It may be pointed out that yield potential is the yield of a crop grown in an environment to which it is adapted and is provided with sufficient nutrients and water, in addition to other stresses being effectively controlled. Thus, considerable yield increases are possible by improving one or more physiological or morphological traits of crop, which in turn are dependent upon the availability of essential nutrients (Evans and Fischer, 1999). Obviously, all these were provided by the wastewater. Nodule number and nodule fresh weight, as well dry weight, was increased under wastewater (Experiments I-V). As pointed out earlier, with the increased amount of nutrients in the medium roots had a better chance to exploit them. This could not only result in increased root proliferation but also nodulation. The positive correlation between leaf N, P and K and nodule number (Table 66) further confirms the assumption. Franco (1977) has cited several authors who obtained increased nodulation and N2 fixation in legumes by utilizing optimum amounts of N in the medium. Similarly, frequent supply of additional P and K in the wastewater also play an important role in enhancing nodulation. In this connection the review by Andrew (1977) gives support to this report of the effect of P contention. Moreover, the importance of K for tropical legumes, specially in N2 fixation by increasing either nodulation or nodule productivity (Duke et ai, 1980) further strengthens our assertion. Add to it the role of Ca (Table 5) in symbiotic N2 fixation (Lowther and Loneragan, 1968; Freire, 1977) and the picture becomes brighter. Increase in NRA was observed throughout the course of study (Tables 18, 39, 58; Figs. 7a & 14a). The presence of nitrate-nitrogen in the irrigation waster (TPPW) as recorded in Table 5 could be mainly responsible for it. NR is a substrate-dependent enzyme (Afridi and Hewitt, 1964; Campbell, 1999). The positive regression observed between leaf N content and NRA (Figs 17-21) buttresses this view. After absorption by roots the N was translocated to leaves (Tables 20, 40 &. 60), which is a major site for its reduction. NRA seems to be indirectly affected by the presence of P in wastewater. P is involved in phosphorylation and diversion'simple towards respiration as a resuh of which oxidation of photosynthates produces more reducing power subsequently for nitrate-mediated NOa' reduction. In comparison to N and P, K

105 Table 66. Correlation coefficient values (r) between the nodule number with leaf NPK contents and seed yield at three stages (Experiments I-V)

Nutrients/ Nodule number Experiments seed yield Stages Vegetative Flowering Fruiting N 0.995** 0.985** 0.998** P 0.965** 0.994** 0.992** K 0.954** 0.965** 0.953** I Seed yield 0.998** 0.999** 0.998**

N 0.996** 0.998** 0.995** P 0.970** 0.976** 0.981** K 0.970** 0.981** 0.989** II Seed yield 0.998** 0.995** 0.998**

N 0.958** 0.961** 0.957** P 0.808* 0.913** 0.837** K 0.920** 0.947** 0.853** III Seed yield 0.994** 0.998** 0.994**

N 0.957** 0.959** 0.885** P 0.897** 0.905** 0.823** K 0.936** 0.929** 0.891** IV Seed yield 0.990** 0.991** 0.976**

N 0.995** 0.999** 0.999** P 0.998** 0.996** 0.997** K 0.997** 0.959** 0.998** V Seed yield 0.999** 0.999** 0.999** »3U - (a) • y == 222.59X -112.41 ^ 800- R^ == 0.8268 Ic ^ 750- • '60 •S 700- • e •^ 650 - < • ^^='^ "• ^ 600 -

550 - 1 1 1 —1 3.2 3.4 3.6 3.8 4.2 Nitrogen content (%) 750 y = 230.02X - 5.4374 (b) -"" 700 R^ = 0.7883

_ 650 H '60 O s 600-

550-

500 —I— 2.2 2.4 2.6 2.8 3.2 Nitrogen content (%) 650 y = 309.09.\ +23.794 (c) ~r 600 R^ = 0.795

_ 550 '60 O 6 500

450

400 —I— —I— —I— —I— 1.3 1.4 1.5 1.6 1.7 1.8 1.9 Nitrogen content (%)

Fig. 17 Linear regression between NRA and nitrogen content at (a) vegetative, (b) flowering and (c) fruiting stages computed from the data in table 18 and 20. 700 y=192.14x-99.351 (a) R^ = 0.8844 -r 650

- 600

i 550

500

450 —I 1— 3.0 3.2 3.4 3.6 3.8 4.0 Nitrogen content (%) 600 (b) 580 y = 216.91x-61.989 ~r 560 A R^ = 0.8749 ^ 540- - 520- 60 o 500- i 480 J 460 g 440 420 400 —I— 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3.0 Nitrogen content (%)

1.8 Nitroge content (%)

Fig. 18 Linear regression between NRA and nitrogen content at (a) vegetative, (b) flowering and (c) fruiting stages computed from the data in table 18 and 20. •3

4.9 5.1 Nitrogen content (%) 900 y=151.65x +304.94 850 - R' = 0.8559

^ 800 U-, '60 "o 750 S =3 700 g< 650

600 —I— 2.3 2.5 2.7 2.9 3.1 3.3 3.5 3.7 3.9 Nitrogen content (%) 750 y = 267.09x+184.86 (c)

-'^ 700 R^ = 0.8675 bu _ 650 £ a. 600 < 550

500 —r— —I— —I— —I— —r— —I— 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2.0 Nitroge content (%)

Fig. 19 Linear regression between NRA and nitrogen content at (a) vegetative, (b) flowering and (c) fruiting stages computed from the data in table 38 and 40. 575 —1 1 1 1 —1 r- 3.2 3.4 3.6 3.8 4 4.2 4.4 4.6 4.8 Nitrogen content (%)

2.6 2.8 Nitrogen content (%)

(c) y== 210.24x-t-162.94 560- • • R^ = 0.8751 540- 520 - ^^^ •

"60 500- "o e 480- a. ••^ 460- 440- ^^...-.^^'^ • 420 -

400 - T 1 r 1 —1 1 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 Nitroge content (%)

Fig. 20 Linear regression between NRA and nitrogen content at (a) vegetative, (b) flowering and (c) fruiting stages computed from the data in table 38 and 40. 1050 y=216.04x +23.509 ^ 1000 R^ = 0.9948

950

"o 900 E 850 <

800

750 3.3 3.9 4.9 Nitrogen content (%) 900 875 - y = 255.48x +39.864 _r^ 850 - R^ = 0.9988 > 825 fc 800 - "M 775 - 1 750- 3 725 ^ 700 Z 675 650

625 1 1 1 1 1 1 1 1 1 1 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3.0 3.1 3.2 3.3 3.4 Nitrogen content (%) 750 y = 377.06X + 33.285 725 R^ = 0.9994 700 675 650 "S B 625 H 600 575 H 550 525 —I— —I— —I— 1.3 1.4 1.5 1.6 1.7 1.8 1.9 Nitroge content (%)

Fig. 21 Linear regression between NRA and nitrogen content at (a) vegetative, (b) flowering and (c) fruiting stages computed from the data in table 58 and 60. Discussion is proved to be an activator of many enzymes including NRA (Suelter, 1970). Similar to above observations, effect of wastewater was noted to be significant on seed protein content in Experiments III and IV, when graded dose of N were applied, while it was non-significant in I, II and V (Figs. 9c & 16c; Table 25, 45 & 65). Increase in seed protein content in Experiments III and IV was due to the presence of N, P and K in wastewater (Table 5) which might have played their roles as explained on page 111 and 113, respectively. The data reveals that protein content of seeds of the plants grown with wastewater was at par with the seed protein content of those grown under GW. This was also observed earlier at Aligarh by Aziz et al. (1999) while working with petrochemical refinery wastewater. The reason may be traced to the "dilution factor". Because of increased seed yield, apparently due to enhanced se^d production in wastewater treated plants. Thus, the tendency to cross the level of significance was nullified by the dilution effect. To confirm this assertion, the seed yield obtained with wastewater was computed with seed protein percentage which showed a marginal increase in protein yield plant"' over the ground water- irrigated plants. However, it may be inferred that wastewater has deleterious nor beneficial effect on seed quality. This may still be considered as a plus point for wastewater irrigation of chickpea. In all the five experiments, shoot length (Tables 6, 26 & 46; Figs. 3a «fe 10a), shoot fresh weight (Tables 7, 27 & 47; Figs. 3b & 10b) and dry weight (Tables 8, 28 & 48; Figs. 3c & 10c), root length (Tables 12, 32 & 52; Figs. 5a & 12a) and root fresh weight (Tables 13, 33 & 53; Figs. 5b & 12b) and dry weight (Tables 14, 34 & 54; Figs. 5c & 12c) increased progressively upito the fruiting stage which is a common phenomenon among various cereals and pulses. Considering nodule formation and growth, nodule number (Tables 15, 35 & 55; Figs. 6a & 13a), nodule fresh weight (Tables 16, 36 & 56; Figs. 6b & 13b) and dry weight (Tables 17, 37 & 57; Figs. 6c & 13c) was noted to increase only uplo the following stage and decreased thereafter. It may be due to the fact that initially the competition for photosynthates was confined to roots, nodules and aerial vegetative organs, but when flowering and fruiting started, these new sinks might have provided more demanding sites for the photosynthates, thus creating shortage for the nodules as a result of which their degradation set in. It is

106 Discussion

noteworthy that chlorophyll and NRA also decreased with increasing age of the plants, comparatively more slowly from vegetative to flowering stage and more sharply from flowering to fruiting stage. This may by attributed to the mobilization of organic and inorganic substances to sink. The concomitant increase in leaf area would thus result in further dilution. This confirms that the density of the photosynthetic pigments (chlorophylls) and the enzymes (NR) unit'" leaf area becomes lesser and lesser with advancing age as pointed out by Hesketh et al. (1981), Bhagsari and Brown (1986), Davies et al. (1987). Among the three major nutrients (N, P and K), assayed in leaves, K content was highest (Tables 22, 42 & 62; Figs. 8c & 15c), followed by that of N (Tables 20, 40 & 60; Figs. 8a & 15a) and P (Tables 21, 41 & 61; Figs. 8b 8c 15b) in that order. For higher plants K is the only essential monovalent cation among the macro-nutrients and generally it is also the most abundant cation in plant tissues (Hubber, 1985). Further, like chlorophyll and NRA, leaf N, P and K contents also decreased with increase in age of the plants. Mention may be made of the work of Rhykerd and Overdahl (1972) wherein rapid decline in leaf K concentration has been reported with maturity in forage legume herbage. Similarly, decline in leaf P concentration with growth was observed by Gomide et al. (1964) in six tropical grasses. They were of the opinion that this decline was due to the dilution factor because of increased growth and/or redistribution of these nutrients to younger plant parts. Like P and K, the observed decreased in N was due to exponential increase in growth (weight and volume) due to which any increase in nutrient concentrations is nullified and even higher quantities of nutrients appear to be less when expressed on unit'" basis (Moorby and Besford, 1983). In addition, the translocation of nutrients to sinks (seeds) during their formation could also be responsible for such observations.

Considering the effect of fly ash on growth and yield parameters, including seed significant increase due to 10% fly ash application was noticed, higher levels being less effective (Figs. 22a & b). It has been supported that fly ash can increase the soil fertility by improving its texture (Chang et al., 1989) and water holding capacity (Sharma et al., 1990), thereby affecting the plant growth indirectly. Its most important direct role is to correct the nutrient balance in the medium (Hill and Lamp, 1980) as

107 o en 'tn C3 X)

•*-» C o I > TD C

c .E < c

> c

KS

c

'C

X

O O 0) OQ g^ 1

1 • 1 I 1 N o 00 VO c5 ( juBid 3) piaiX paag ( }UB|d 3) pfsiX paag o __

oi (N Discussion

some of the naturally existing essential nutrients enrich it (Klein et ai, 1975; Koakinen et al., 1975). It is known to be source of B (Wallace and Wallace, 1986), Ca (Martens and Beahm, 1976), Cu (Wallace et ai, 1980), K (Martens et al, 1970), Mg (Hill and Lamp, 1980), Mo (Gary et al, 1983), S (Elseewi et al, 1978) and Zn (Schnappinger et al, 1975). Expectedly, it was due to the presence of these essential element in our fly ash samples (Table 4) that supplemented those supplied by the soil and wastewater. However, the benefit of fly ash proved only of limited nature as noted above. The decrease in yield was probably due to increased levels of sulphate, chloride, carbonate and bicarbonates (Table 4). Some toxic compounds i.e. dibenzofuran and dibenzo-p-dioxine mixture (Helder et al, 1982; Sawyer et al, 1983) and elements like Ni, As, Cd, Cr, Pb, Se, Zn, Cu (Wadge and Hutton, 1987) were reported to occur in fly ash might have also contributed towards the lesser yield under higher fly ash concentrations. Detrimental effect of higher levels of fly ash on plants have also been reported earlier due to either the phytotoxicity of B (Adriano et al, 1978) or a shift in the chemical equilibrium of the soil (Singh and Yunus, 2000). Nodulation, like growth and yield, was increased on adding fly ash albeit upto a limited level (10%). More than 10% amendment decreased it due to variation in pH. At higher levels, toxic amounts of soluble salts released from fly ash seem to affect roots and rhizosphere adversely. It may also be added that high doses of fly ash added to the soil decrease the microbial activity due to change in soil salinity or concentrations of potentially toxic elements (Singh and Yunus, 2000). This could not only delay nodulation but also cause a decrease in their number as noted by Martensson and Witter (1990). NRA and leaf N, P and K were also decreased by higher doses of fly ash (Experiments I & II). Although fly ash contained an extremely small amount of nitrogen, an increase in NRA by its application was observed in the present study. The presence of Mo (Gary et al, 1983) in fly ash and sufficient quality of available nitrogen in the soil (Table 4) might have accelerated the rate of NR activity. Gonsidering the increase in seed protein content due to the application of fly ash (Table 25; Fig. 9c) the pressure of additional P and K in it may be responsible for it. This is clear from regression studies of the two nutrients and seed protein (Figs. 24- 32). This has also been reported by Bhaisare et al (2000), Khan et al (1996),

108 25.0 - (a) y = 21.42X+15.153 ^ 24.5 - R^ = 0.7451 ^^---^ ^ 24.0 - • ^ 1 23.5 - • ^^^^ c • S 23.0 - 2 a. •ua 22.5 -

" 22.0 - •

21.5- T 1 1 1 1 1 0.31 0.33 0.35 0.37 0.39 0.41 0.43 0.45 Phosphorus content (%) 25.0 (b) y = 28.927x+16.171 ^^ 24.5 % R^ = 0.6519 *i 24.0 t) o 23.5 c 2 23.0 8 o< •a 22 5 u C/«1 22.0

21.5 —I 1 1— 0.2 0.21 0.22 0.23 0.24 0.25 0.26 0.27 0.28 0.29 Phosphorus content (%) 24.5 y = 33.582x+16.514 ^ 24.0 R^ = 0.63 I 23.5 - o c 23.0

1. 22.5 •o u u ^ 22.0

21.5 —I— —I— 0.16 0.17 0.18 0.19 0.2 0.21 0.22 0.23 0.24 Phosphorus content (%)

Fig. 24 Linear regression between seed protein content and phosphorus content at (a) vegetative, (b) flowering and (c) fruiting stages computed from the data in tables 21 and 25. 3.5 3.7 3.9 4.1 4.3 4.5 4.7 Potassium content (%)

2.4 2.5 2.6 2.7 2.8 2.9 3.1 3.2 3.3 Potassium content (%)

2.3 Potassium content (%)

Fig. 25 Linear regression between seed protein content and potassium content at (a) vegetative, (b) flowering and (c) fruiting stages computed from the data in tables 22 and 25. 21,6 y= 16,953x+14.641 (a) 21.4 ^^% R^ = 0.7648 • 21.2 a 21.0 e o 20.8 (J c 20.6 20.4 • ^^^ 20,2 u o 20.0 ^^^ •• 19.8

19.6 1 1 1 1— 1 0.30 0.32 0.34 0.36 0.38 0.40 0.42 Phosphorus content (%) 21.6 (b) 21.4 y = 25.738.\ + 14.867 ^^^* • 21,2 R- = 0.744 21.0 u s 20.8 o u 20.6 c • ^,^'^ s 20.4 o< 20.2 •ua u 20.0 CO ^^-^^^ •• 19.8

19.6 - 1 1 r 1 1 1 0.19 0.20 0.21 0.22 0.23 0.24 0.25 0.26 Phosphorus content (%) 21.6 (c) 21.4 y = 32.942.\ + 14.933 21.2 R- = 0.7019 «-* 21.0 uB o 20.8 (J .c 20.6 '5 e 20.4 20.2 Tu3 20.0 19.8 19.6 0.14 0.15 0.16 0.17 0.18 0.19 0.20 Phosphorus content (%)

Fig. 26 Linear regression between seed protein content and phosphorus content at (a) vegetative, (b) flowering and (c) fruiting stages computed from the data in tables 21 and 25. 3.7 3.9 4.1 4.5 Potassium content (%) 21.6 21.4 y = 2.3061x4-14.564 21.2 R^ = 0.755 21.0 H I 20.8 20.6- 20.4- 1 20.2 20.0 19.8 19.6 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 3.0 3.1 Potassium content (%)

1.95 2.05 2.15 2.25 2.35 2.45 2.55 Potassium content (%)

Fig. 27 Linear regression between seed protein content and potassium content at (a) vegetative, (b) flowering and (c) fruiting stages computed from the data in tables 22 and 25. 24.0 y = 16.488X + 16.091 23.5 R^ = 0.9597

s 23.0 o u c 22.5 'S s 22.0 a, •o o o 21.5 C/2 21.0 0.30 0.33 0.35 0.38 0.40 0.43 0.45 0.48 0.50 Phosphorus content (%) 24.0 (b) y = 27.325X + 16.689 • /^f ^ • 23.5 R^ = 0.9617

I 23.0 c o u c 22.5 • • •^ "3 G. 22.0 ^^ • •a u ^ 21.5 H • 21.0 1 —1 —1 —1 —1 0.16 0.18 0.20 0.22 0.24 0.26 0.28 Phosphorus content (%) 24.0 (c) y = 32.453x+17.334 23.5 R^ = 0.9304

23.0

22.5

22.0 u 21.5 H

21.0 -r -r 1 1 1 1 0.12 0.13 0.14 0.15 0.16 0.17 0.18 0.19 0.20 0.21 Phosphorus content (%)

Fig. 28 Linear regression between seed protein content and phosphorus content at (a) vegetative, (b) flowering and (c) fruiting stages computed from the data in tables 41 and 45. 24 (a) y= 1.8588X+14.447 23.5 R^ = 0.9886

g 23 oc u c 22.5 1 a. 22 •a o u 21.5

21 —I— —I— —r- 3.5 3.7 3.9 4.1 4.3 4.5 4.7 4.9 5.1 Potassium content (%)

2.7 2.8 2.9 3.2 3.3 Potassium content (%) 24.0 y = 3.6496x+13.852 R' = 0.9971

2.8 Potassium content (%)

Fig. 29 Linear regression between seed protein content and potassium content at (a) vegetative, (b) flowering and (c) fruiting stages computed from the data in tables 42 and 45. 4.2 4.6 4.8 Nitrogen content (%) 25.8 25.6 y=1.7605x+19.831 R^ = 0.7644 25.4 H 25.2 25.0 H 24.8 e 24.6 a, •a u 24.4 u to 24.2 H 24.0 2.3 2.7 2.8 2.9 3.0 Nitrogen content (%) 25.8 (c) 25.6 . y = 3.1908X+19.756 R^ = 0.766 25.4 B 25.2 iSJ B O 25.0

Fig. 30 Linear regression between seed protein content and nitrogen content at (a) vegetative, (b) flowering and (c) fruiting stages computed from the data in tables 60 and 65. c U So o c

•Tu3 U CO

0.40 0.42 0.44 0.50 Phosphorus content (%)

0.18 0.19 0.20 0.21 0.22 0.25 Phosphorus content (%) 25.8 (c) 25.6 y = 30.523x+19.83 R^ = 0.7823 2? 25.4 25.2 25.0 H 24.8 24.6 uo 24.4 lO 24.2 24.0 0.13 0.14 0.15 0.16 0.17 0.18 0.19 0.20 Phosphorus content (%)

Fig. 31 Linear regression between seed protein content and phosphorus content at (a) vegetative, (b) flowering and (c) fruiting stages computed from the data in tables 61 and 65. 25.8 y= 1.087X+19.834 25.6 R- = 0.8115 25.4 H 25.2 25 24.8 24.6 T3

C/3 24.4 24.2 24 3.8 4.8 5.4 Potassium content (%) 25.8 25.6 y = 2.1951x+18.428 R^ = 0.9445 25.4 C o 25.2 c o u 25.0 e 24.8 H 24.6 u o 24.4 H to 24.2

24.0 1 2.5 2.8 2.9 3.0 3.1 3.4 Potassium content (%)

—r— 2.3 2.4 2.5 2.6 2.7 2.8 Potassium content (%)

Fig. 32 Linear regression between seed protein content and potassium content at (a) vegetative, (b) flowering and (c) fruiting stages computed from the data in tables 62 and 65. Discussion

MiLovsky (1992), Sriramachandrasekharan (2001). Similarly, due to phytotoxicity of some heavy metals and conversion of some trace elements like Mo and B into some inorganic complexes availability of nutrients including NPK was adversely affected under high levels of fly ash (Bilteanu et ai, 1973). Let us now consider some of the data of Experiments III &. IV. Plant growth is the expression of interplay between meristematic activities and metabolic processes leading to an increase in biomass (Moorby and Besford, 1983). In addition to its role in cell division and expansion (Gardner et ah, 1985), N is also essential for a number of biologically important molecules. Therefore, the requirement of N (and the other essential nutrients) during the vegetative growth of a plant is determined primarily by the rate of CO2 assimilation and if it is high, the required nutrients must be correspondingly at optimum levels in order to convert the photosynthates efficiently into other metabolites. Thus, growth and yield parameters were noted to be significantly affected by N application (Nio proving optimum) as a resuh of the cumulative enhancement of growth and yield parameters, including seed yield (Figs. 22a & b). This dose was also found to be optimum for leaf area, NRA and chlorophyll content (Tables 30, 38 & 39; Figs, lib, 14a & b) which finally led to more pods and the heavier seeds (Table 43; Fig. 16a). By contrast application of excess N (N30) resulted in decreased grain yield and proved deleterious. Toxicity due to N, when applied as urea is known to appear at two stages of plant growth. The first at seedling stage may be due to accumulation of NH4* (i-^^r hydrolysis of urea) which becomes toxic at pH 8 and above. The second is due to accumulation of NO2 under certain conditions damages young plants (Court et ai, 1964). Contrary to above findings, higher optimum doses upto 30 kg N ha'' were reported for chickpea by Sharma et al. (1989) and Krishna et al. (2001). It was not surprising that in our study comparatively lower dose (Nio) proved effective as the applied wastewater had sufficient N in the form of NH4"' and NO3" ions. In case of legumes due to rhizobial activities, host plants grow well in soil even with low N doses and no benefit from this association may occur if high levels of fertilizer N are given (Ozanne, 1980). Which was in conformity with the present observations. Since Njo and N20 were at par in their effect therefore it may be concluded that N20 led to

109 Discussion

luxury consumption, thereby proving wasteful, while N30 affected adversely thereby proved toxic when wastewater was the source of irrigation, which proved economically as well as environmentally viable. When nodulation was considered similar observations were made. The beneficial effect of lower dose (Nio) was noted to increased root formation (Table 32; Fig. 12a). This provided more surface area for bacterial infection. However, application of N beyond a certain level is known to delay and even suppress nodulation (Wilson and Hallsworth, 1965; Arrese-Igore/a/., 1997; Koike e/a/., 1997; Krugova, 1997; Nishiwaki et al., 1997). On the other hand, the crop grown without nitrogen (No) expectedly gave significant lowest values as some starter dose of N is always needed even by the leguminous plants to grow normally. Nitrate reductase levels have been shown to fluctuate in response to changes in environmental conditions, including availability of N (Afridi and Hewitt, 1964; Beevers and Hageman, 1972). Enzymes are sensitive to nutrient levels as is indicated in the present study where NRA was found to decrease with comparatively higher N dose (Table 38; Fig. 8a). Similar observation has been made in trifoliate leaves in Phaseolus lunatus at different canopy positions by Wallace (1986) and Andrews et al. (1990). The induction of NRA requires very low concentration of nitrate suggesting that nitrate is actually sensed more as a hormone than as a nutrient (Crav^ord, 1995). Nitrogen also increased the leaf chlorophyll and NPK contents as it increased the availability of substrate for protein synthesis allowing the development of more and larger chloroplasts with extensive thylakoid system and larger stomal volme (Kutik et al, 1995). The increase in leaf NPK (Tables 40, 41 & 42; Figs. 15a, b, c) was due to the synergistic interplay of the three nutrients, which are known to accelerate root proliferation, thus,^ extracting more nutrients present in the root zone leading to development of larger canopies (Table 30; Fig. lib) and greater dry matter accumulation (Tables 28, 34, 37 &. 43; Figs. 10c, 12c, 13c & 16b). Similar positive interactions between N and P were also noted by Russell (1973) and between N and K by Murphy (1980). N as an essential macronutrient has the distinction of being absorbed both as cation as well as an anion. This puts N in a unique relationship of both an anion-cation as well as cation-cation interaction.

110 Discussion

Expectedly the application of N enhanced seed protein contents (Figs. 23c & d) as it chief constituents of proteins. Its adequate supply can increase the amino acid levels through the conversion of organic produced from carbohydrates during respiration. As pointed out by Pretty (1980), some quality factors in a few grasses were related to the effective utilization of N and the conversion of N-compounds into true proteins. Improvement in seed protein content was also boosted due to the addition of K, applied uniformity as the starter dose alongwith N, as K influences the level of some non-protein N components and positive role in converting these proteins. The N effect on seed protein was also dependent upon the type of crop, its cultivars and other environmental factors including water. Smika and Greb (1973) observed the relationship of soil NOa'-N and soil water for the protein in wheat. The former was positively correlated with grain protein where opposite relationship was noted due to available soil water. In their opinion adequate soil moisture in addition to N were the important factors for this parameter. Since, the present work was carried Wut in pots and water was given regularly, therefore, possible protein in the present study was increased. Growth and yield of chickpea also responded favaourably to P application (Experiment V). It has to be kept in mind that plant factors as well as soil factors have great influence over the utilization of this indispensable nutrient. Regarding the former, root structure, species differences and the effect of defoliation are considered important. Among the soil factors, phosphate buffering capacity, distribution of P in soil profile, reactivity of soil applied phosphate fertilizer, soil pH and the residual value earlier applications, in addition to moisture regime and presence of other nutrients, play important roles for its availability. Its effect on leaf area enhancement has been reported by Rao and Subramanian (1990) in cowpea and by Reddy et a/. (1991) in groundnut. Similarly, increase in branch number has been noted due to P apphcation in chickpea by Parihar (1990), in dry matter accumulation by Khokar and Warsi (1987), in nodule number and nodule dry weight by Idris et al. (1989) as well as Singh and Ram (1992) and in seed yield by Parihar (1990). Legumes show an evident preference for phosphatic fertilizers (Raju and Verma, 1984). In the present study, P30 proved optimum for most of the growth, yield (Fig. 22c) and physiological

III Discu.s.sioii

parameters (Tables 58-62). As mentioned earlier, the enhanced leaf area (Table 50) enabled the plants to produce more photosynthates and dry matter (Tables 48, 54, 57 &. 64) and also more pods (Table 63). These observations are pertinent, as P is known to facilitate the partitioning of photosynthates between source and sink (Giaquinta and Quebedeaux, 1980). We may now consider the role of P in nodulation and the resulting increase in yield. It has been observed earlier that dinitrogen fixation by Rhizobium is enhanced if the plants are supplied P along with K (Gukova and Tjulina, 1968; Wu et al., 1969; Mengel et al, 1974). Mention may also be made of the observations by Andrews and Robins (1969) who noted a positive correlation between applied P and leaf N concentration while working on tropical and temperate pasture legume. They further reported that P fertilization specially on N and P deficient soils, enhanced nodule development by increasing nodule number, dry weight and nodule growth rate. In the present study, the observed increase in nodulation due to P application (Tables 55-57) was through its role in the proliferation of roots which provided larger surface area as indicated by higher root dry weight (Table 54) of the P-treated plants. This favourable response regarding dry matter accumulation was probably due to increased production of photosynthates in the shoot and its transfer to root. This assumption is strengthened by the observed increase in leaf area (Table 50) and shoot dry weight (Table 48). In our study, P30, a dose comparatively lower than that recommended for chickpea, seems to have become sufficient probably due to the presence of additional P specially in wastewater and to a lesser extent in fly ash. It may be pointed out that P is often limiting nutrient due to its low availability in comparison to K which is not only easily recycled fi:om organic residues but is also reading available from the fertilizers applied to the soil. Hence, the ameliorating role played by the P present in wastewater and fly ash. A deficiency of N, on the other hand, is largely compensated in the legume corps through nitrogen fixation. Therefore, the quality of P fertilizer to be applied to such crops is critically important as the amount of available P often declines with time. In addition, P is neither recycled like sulphur nor is readily released from organic residues. Therefore, the regular supply of P, through wastewater application, proved effective and useflil as mentioned above. P also

112 Discussion

increased leaf N content but it remained ineffective in increasing leaf P level. Such observations of enhanced N concentration due to P fertilization in tropical legumes were also reported by Shaw et al. (1966), Andrews and Robins (1969) and Dradu (1974). Higher nodulation due to P application might have increased the N content in leaf through more efficient nitrogen fixation, followed by that of K, thus indirectly enhancing the growth performance of the corp. The application of P also proved beneficial for seed protein content (Table 65; Fig. 23 c) due to its assured availability and continuous utilization by carbon skeletons for amino acid synthesis as well as that of energy rich ATP and manifesting this in enhanced protein synthesis in the seeds. Being a part of the protein molecules enhanced N levels in leaves due to the application of P (Table 60) might have triggered and maintained the conversion of various organic acids (produced from carbohydrates during respiration) into amino acids. The known role of K (also present in sufficient quantity) in activating various enzymes involved in protein synthesis (Evans and Sorger, 1996; Tamhane et al., 1970). The strong positive regression of seed protein content with leaf NPK contents (Experiment V) at various stages of growth further strengtheqf this view (Figs. 30-32). The ameliorative effect of nutrients present in the applied wastewater and fly ash, together with the N and P applied as fertilizers, was pronounced when interaction was considered. On defining interaction, Russell (1973) states that if two factors are limiting or nearly limiting growth, adding only one of them will have little effect, while adding both together will have a very considerable effect. In the context of crop plants, two such factors show a positive interaction if the response of the crop to both together is larger than the sum of responses to each separately. If the crop response to the two factors together equalled the sum of its responses to each separately, we could say that the two factors showed no interaction, or worked entirely independently of each other. On the other hand, if the response to the two factors together is less than the sum of the responses to each factor separately, they should be deemed to have a negative interaction with each other. In this context, mention may be made of the work conducted by Bouldin and Sample (1958, 1959) who observed that such a positive interaction was also due to the increased of P because of close

113 Discussion association of the two fertilizers applied as sources of N and P. Reports are also available where P has been found to increase N concentration in same tropical legumes (Andrew and Robbins, 1969; Dradu, 1974). Dradu (1974) also observed depressed nodulation, seedling vigour and seedling growth in plants grown without P. Similarly Kilcher et al. (1965) observed that N dose had little effect on yield increases while the combination of N and P increased the yield in forages. Similarly, Black (1968) observed that N and P together increased the forage yields in wheat grass and no yield increase was observed under P alone. It may be emphasized that wastewater and fly ash supplemented these nutrients thereby proving economically efficacious on the one hand and environmentally acceptable on the other. Among the significant interactions, low concentration of fly ash plus wastewater i.e. TPPW x FAio (Experiments I & II) proved beneficial due to the positive nutritional role played by the constituents of the wastewater products generated from the same source (Thermal Power Plant). Whereas, addition of nitrogen, TPPW x FA20 >< Nio (Experiments III & IV) and phosphorus, TPPW x FAio x Nio '^ P30 (Experiment V) further improved the performance of the crop tested. As mentioned earlier fly ash was deficient in N which was amply compensated by the application of wastewater having sufficient nitrogen in the form of NHt"^ and NO3" in the presence of low doses of N and P fertilizers. Crop species differ in their morphological and physiological characteristics as well as yielding ability in response to their surroundings. Genetic variability is supposed to be largely responsible for such observations (Frageria et al., 1991) although environmental factors do play a role. In agricultural crops, therefore, genetic potential must be of sufficient magnitude and flexibility so that they may be grown over a wide range of agroclimatic conditions. This accounts for differences in crop productivity and simultaneously allows a particular crop or cultivar to adapt itself to a particular environmental conditions (Lafever, 1981; Heinrich et al., 1983; Bruckner and Frohberg, 1987). The pooled data of Experiments I and II shows that for vegetative growth, wastewater proved beneficial for both cultivars (Avarodhi and BG- 256) whereas in Experiments III and IV, BG-256 performed better. Under fly ash treatments (Experiments I and II) BG-256 performed better than Avarodhi as growth of the former was enhanced by fly ash in general and FAio in particular. Similarly,

114 Discussion

Nio (Experiments III and IV) significantly enhanced the growth of BG-256. The percent investigation, thus showed that due to the superior inherent genetic potential of BG-256, it proved more efficient in comparison to Avarodhi. However, it must be admitted that the amount and kind of nutrient applied for better growth and yield in particular crop, and even its species or cuhivars, is important as the magnitude of differences various between the species well adopted to the same climate, same soil and in some cases even the same management. The superior performance of BG-256 was due to increased nodulation, and dry weight as well as better developed root system (Tables 15-17 & 35-37; 8, 28, 48, 12-14, 32-34) in comparison to Avarodhi. It also showed enhanced leaf area resulting into higher matter accumulation (Tables 8, 28, 14, 34, 17, 37, 23, 43). The pooled analysis (Experiments I-IV) show that BG-256 significantly differed from Avarodhi in leaf NRA, chlorophyll content, leaf NPK contents, seed yield (Fig. 22) and seed protein contents (Fig. 23) under wastewater, fly ash and nitrogen. The better performance of BG-256 in this regard could be traced back to enhanced shoot dry weight and root dry weight - the most important criteria to access vegetative growth. The differences in N, P and K status of the two cultivars reflected their differential efficiency to absorb and accumulate these nutrients (Tables 20-22, 40-42). To conclude, the addition of P in combination with other inputs (TPPW x FAio ^ Nio X P30) significantly increased growth, yield (Fig. 22) and nodulation in chickpea cv. BG-256 which ultimately led to higher pod number, heavier seeds (Table 63) and higher seed protein content (Table 65). This improvement was due to the combined effect of N and P applied to the soil together with the nutrients present in wastewater and fly ash, confirming their suitability as a combined source of irrigation water and nutrients (Tables 4 & 5). These findings thus provide a positive conclusion with regard to the objectives of the present study. Therefore, for the cultivation of chickpea, basal application of 10 kg fly ash ha', 10 kg N ha' and 30 kg P ha'" may be recommended under TPPW irrigation. Among the available varieties, BG-256 may be preferred for cultivation in this region (Western Uttar Pradesh, India). Finally, TPPW and fly ash, which are by all means waste product of Thermal Power Plant, may be profitably utilized for agriculture purpose.

115 a (U

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c « E ?- ^- /^^ < a. W ^—^ c 4-* C R (N o U. Q u CQ c «+- 4-* C uo. E C o. i<^ ton T3 VO Vl CO u CN 1 a. t/3 o S 'S CQ < o JC -t—• m 1 (N r~ in

Conclusion 1. Thermal Power Plant Wastewater (TPPW) irrigation proved beneficial for growth, nodulation and yield of chickpea (Experiments I-V) and may, therefore, be recommended for the purpose of irrigation of this crop. 2. TPPW, on its own, neither increased nor decreased seed protein content. This may be taken as a positive point for its use for such crops. However, it is noteworthy that when TPPW was supplemented with fertilizer N (Experiments III and IV) seed protein content was increased upto 4%. 3. Study of wastewater revealed its suitability for irrigation as the analysed parameters were within the permissible limits of India Standards. 4. Inclusion of 10% fly ash (FAio) in the irrigation water proved beneficial in comparison to FA20 and FA40 (Experiments I and II). 5. Since fly ash was deficient in N, its application should be restricted to crops that have the ability to fix atmospheric nitrogen. 6. Application of fly ash (FAio) increased nodulation and seed protein content but it showed decreasing trend as its level was increased (Experiments I & II). 7. Plants irrigated with TPPW performed better when supplemented with low fertilizer N level, Nio (Experiments III & IV), thus proving the utility of wastewater in saving some amount of costly nitrogenous fertilizers which simultaneously solving the problem of its disposal partially. N30 proved deleterious, while N20 showed luxury consumption when given with wastewater. 8. Nodulation and seed protein content were increased by the application of Nio, while N30 decreased nodulation (Experiments III & IV). 9. Low P levels (Pio and P20) proved efficient for most of the parameters while P30 proved optimum (Experiment V). 10. Nodulation, as well as protein content, increased with increasing levels of P. 11. Among the nutrients, K was accumulated more by the leaves, which was followed by N and P. 12. Shoot length, shoot fresh and dry weight, branch number, leaf number, leaf area, root length, root fresh and dry weight increased with increasing age of

116 Discussion

the plants. 13. NRA, chlorophyll and leaf N, P and K contents decreased with increasing age of the plants. 14. Nodule number, nodule fresh weight and dry weight increased only upto flowering stage and decreased at fruiting. 15. Both BG-256 and Avarodhi cultivars grew well under wastewater irrigation but BG-256 responded better and may, therefore, be preferred over the latter for cultivation under wastewater irrigation together with FAio added to a low fertilizer dose (N10P30K20) as this combination of TPPW, FA and fertilizer proved highly cost-effective. Proposal for future studies The above observations recorded meticubsly in pot culture and spread over a three year period have no doubt established the utility of wastewater and fly ash for growing chickpea. Some economy of fertilizers has also been noted. However, before recommending the technique to the local farmers for adopting, the test should be repeated in farmer's fields for verification. In addition, an attempt may be made to record the following observations that could not be undertaken in the present study due to unavailability of facilities: 1. Microbiological study of wastewater. 2. Heavy metal analysis in wastewater, fly ash, soil and seeds. 3. Nitrogenase activity for assessing the N2 fixing ability of chickpea and other seed legumes under wastewater and fly ash application.

117 Summary Chapter-6 SUMMARY

Chapter 1 (Introduction) explains and justifies briefly the need for undertaking the present study. Chapter 2 (Review of Literature) includes an up-to-date review pertaining to fly ash and industrial wastewater, their effect on plants, especially the N2 fixing leguminous crops, and of the fertilizer requirement of chickpea (Cicer arietinum L). Chapter 3 (Materials and Methods) contains the methodology and techniques employed for the five pot trials undertaken together with relevant data regarding soil, fly ash and water analysis. Chapter 4 (Experimental Results) comprises the data analysed statistically and presented in the form of Tables (6-65) and Figures (3-16). The significant data are summarized briefly below. Experiments I & II were conducted on chickpea cultivar BG-256 and Avarodhi respectively during the "rabi" (winter) season of 1999-2000 to study the comparative effect of thermal power plant wastewater (TPPW) and ground water (GW) and four levels of fly ash (0, 10, 20 and 40kg ha'') under uniform basal doses of N, P and K (20kg ha*' each). TPPW proved superior for most of the parameters studied, including nodulation, NRA and seed yield but not seed protein content. Among fly ash treatments, FAio proved more effective than FA20 and FA40. The concentration of leaf K was comparatively higher than that of N and P. However, the contents of the three nutrients decreased with increase in growth. Experiments III & IV were conducted during 2000-2001 on the same two cultivars. Four levels of basal nitrogen (0, 10, 20 and 30kg ha'') were tested with a uniform basal dose of P and K (20kg ha'' each) and FAio. TPPW again proved superior to GW for growth and development, including NRA, nodulation and seed yield plant''. Nio proved optimum for NRA, nodulation, seed yield and protein content, whereas N20 encouraged luxury growth and N30 found to be determined for all parameters except leaf NPK content. In these trials, seed protein content also increased upto 4.49% (Experiment III) and upto 12.3% when the data of this Summary

experiment were pooled with those of Experiment IV). When the data of experiments I & II and of Experiments III & IV were pooled to assess the performance of the two cultivars, it was revealed that BG-256 responded better than Avarodhi to wastewater irrigation. Moreover, BG-256 performed better under FAio (Experiments I & II) and Nio doses (Experiments III & IV). Experiment V was conducted during the "rabi" season of 2001-2002 to evaluate the performance of cultivar BG-256 (selected on the basis of its better performance) grown under the same two irrigation sources and five levels of phosphorus (0, 10, 20, 30 and 40kg ha*') supplemented with 10kg fly ash ha'' and 10kg N ha'' (selected on the basis of Experiments I & III respectively), together with uniform basal dose of K (20kg ha''). As noted earlier, TPPW proved more effective than GW while, among phosphorus doses, Pio and P20 proved deficient, P30 was optimum for NRA, nodulation, seed yield and protein content among other parameters studied, whereas P40 proved wasteful. It may, therefore, be concluded that, with increasing levels of P, nodulation and seed protein content increased with P30 proving optimum. It may also be noted that the combination of TPPW, FAio, Nio and P30 proved most effective. BG-256 performed better than Avarodhi with respect to seed yield as well as seed quality. Chapter 5 (Discussion) includes critical consideration of the experimental results and their regression and coefficient correlations (Tables 6-66; Figs. 3-21 & 24- 32) in the light of research work carried out by other scientists on cuhivated crops in general and leguminous crops in particular. Conclusions have been drawn in the end, credit claimed on their basis for important new additions to literature on the subject and some suggestions incorporated for future work. Chapter 6 (Summary) is the present chapter. It gives a glimpse of the entire study. It is followed by an up-to-date bibliography comprising references cited in the text. An appendix comprising the composition of the reagents used during the experimental work, is also appended.

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146 ^^pendix APPENDIX

l-Ainino-2-naphthol-4-suIphonic acid 0.5g l-amino-2-naphthol-4-sulphonic acid dissolved in 195ml 15% sodium bisulphite solution to which 5ml 20% sodium sulphite solution was added.

Alkali iodide azide reagent 50g sodium hydroxide and 15g potassium iodide diluted to 100ml with double distilled water (DDW). Ig sodium azide dissolved in 4ml of DDW and added to the above solution.

• Ammonium acetate solution (IN) Dilute 57ml glacial acetic acid to 800ml DDW and neutralize to pH 7.0 with concentrated ammonium hydroxide and final volume made upto 1000ml.

• Ammonium chloride-ammonium hydroxide buffer (a) 16.9g ammonium chloride dissolved in 143ml concentrated ammonium hydroxide (b) 1.179g of dissolved EDTA and 0.780g magnesium sulphate dissolved in 50ml DDW. Both (a) and (b) solutions mixed and dilute to 250ml with DDW.

• Ammonium molybdate solution (2.5%) (a) 25.Og ammonium molybdate dissolved in 175ml DDW (b) Add 280ml concentrated H2SO4 to 400ml DDW and cool. Mix the two solutions (a) and (b) and final volume made upto 1 litre with DDW.

• Ammonium purpurate 150mg ammonium purpurate dissolved in lOOg ethylene glycol.

• Conditioning reagent 50ml of glycerol mixed in a solution containing 30ml concentrated HCl + 300ml DDW + 100ml 95% ethyl alcohol and 75g sodium chloride.

• Dickman and Bray's reagent 15g ammonium molybdate dissolved in 300ml warm DDW (about 60°C) cooled and filtered, if necessary. To this, 400ml ION HCl was added and final volume was made upto lOOOml with DDW.

• Diphenylamine indicator 0.5g diphenyl amine dissolved in a mixture of 20ml DDW and lOOml concentrated H2SO4.

• EDTA (0.0IM) 3.723g disodium salt of ethylene diamine tetra acetic acid dissolved in DDW and diluted to 1000ml. Appendix

• Eriochrome black T indicator 0.4g Eriochrome black T grind with lOOg powdered sodium chloride.

• Ferrous ammonium sulphate (0.5N) 196g ferrous ammonium sulphate dissolved in DDW, to this 20ml concentrated H2SO4 was added and the final volume made upto 1000ml.

• Ferrous ammonium sulphate solution (O.IN) 39.2g ferrous ammonium sulphate dissolved in DDW. 20ml of concentrated sulphuric acid was added and volume made upto 1000ml.

• Folin phenol reagent lOOg sodium tungstate and 25g sodium molybdate dissolved in 700ml DDW to which 50ml 85% phosphoric acid and 100ml concentrated hydrochloric acid were added. The solution was refluxed on a heating mantle for 10 hrs. At the end, 150g lithium sulphate, 50ml DDW and 3-4 drops liquid bromine were added. The reflux condenser was removed and the solution was boiled for 15 minutes to remove excess bromine, cooled and diluted upto 1000ml. The strength of this acidic solution was adjusted to IN by titrating it with IN sodium hydroxide solution using phenolphthalein indicator.

• Hydrochloric acid (O.OIN) 0.86ml pure hydrochloric acid mixed with DDW and final volume made upto 1000ml.

• Hydrochloric acid (O.IN) 8.62ml hydrochloric acid mixed with DDW and final volume made upto 1000ml

• Hydrochloric acid (0.2N) 17.24ml HCl mixed with DDW and final volume made upto 1000ml.

• Isopropanol solution (5%) 5ml isopropanol mixed with 95ml DDW.

• Liquid ammonia (1:1) Ammonia having 0.88 specific gravity diluted with equal volume of DDW.

• Manganous sulphate solution 108g manganous sulphate dissolved in boiled DDW and volume made upto 200ml.

• Methyl orange indicator (0.05%) 0.5g methyl orange dissolved in 100ml DDW.

• Molybdic acid reagent (2.5%i) 6.25g ammonium molybdate dissolved in 75ml ION sulphuric acid. To this Appendix

solution, 175ml DDW was added and maintained the total volume 250ml.

• Murexide indicator 0.2g ammonium purpurate grind with lOOg powdered sodium chloride.

Naphthylethylenediamine dihydrochloride (NED-HCI) solution (0.02%) 20mg naphthylethylenediamine dihydrochloride dissolved in sufficient DDW and final volume maintained upto 100ml with DDW.

Nessler's reagent 3.5g potassium iodide dissolved in 100ml DDW to which 4% mercuric chloride solution was added with stirring until a slight red precipitate remained. Thereafter, 120g sodium hydroxide with 250ml DDW was added. The volume was made upto 1 litre with DDW. The mixture was filtered twice and kept in an amber coloured bottle.

Olsen's reagent 42.Og dissolved in 1000ml and DDW. The pH was adjusted to 8.5 with the addition of small quantity of sodium hydroxide.

Phenol disulphonic acid This was prepared by taking 25g pure phenol (CeHsOH, crystal white) in a conical flask (500ml) to which 150ml concentrated H2SO4 and 75ml fuming sulphuric acid were added and kept on boiling water bath for 2 hours. After cooling, it was stored in an amber coloured bottle.

• Phenolphthaiein indicator 0.5g phenolphthaiein dissolved in 50ml of 95% and add 50ml DDW. Add 0.05N CO2 free NaOH solution drop wise until the solution turns faintly pink.

Phosphate buffer (O.IM) for pH 7.5 (a) 13.6g potassium dihydrogen ortho phosphate dissolved in sufficient DDW and final volume made upto 1000ml with DDW (b) 17.42g dipotassium hydrogen ortho phosphate dissolved in sufficient DDW and final volume maintained upto 1000ml with DDW. 160ml of solution (a) and 840ml of solution (b) were mixed for getting phosphate buffer.

Potassium chromate indicator (5%) 5g potassium chromate dissolved in DDW and final volume made upto 100ml.

Potassium dichromate solution (IN) 49.04g potassium dichromate dissolved in 1000ml DDW.

Potassium dichromate solution (0.25N) 12.259g potassium dichromate dissolved in DDW and final volume made upto 1000ml.

lU Appendix

Potassium nitrate solution (0.2M) 2.02g potassium nitrate dissolved in sufficient DDW and final volume maintained upto 100ml with DDW.

Reagent A 0.5% copper sulphate solution and 1% sodium tartarate solution mixed in equal volumes.

Reagent B 50ml 2% sodium carbonate solution mixed with 1ml reagent 'A'.

Silver nitrate solution (0.02N) 3.4g silver nitrate dissolved in DDW and diluted to 1000ml.

Sodium hydroxide solution (O.IN) 4g sodium hydroxide dissolved in 1000ml DDW.

• Sodium hydroxide solution (IN) 4g NaOH dissolved in DDW and final volume made upto 100ml.

• Sodium hydroxide solution (2.5N) lOOg sodium hydroxide dissolved in 1000ml DDW.

• Sodium hydroxide solution (6N) 24g NaOH dissolved in sufficient DDW and final volume made upto 100ml.

• Sodium thiosulphate solution (0.025N) 6.2g sodium thiosulphate dissolved in 1000ml DDW.

• Stannous chloride solution lOg crystalline stannous chloride dissolved in 25ml concentrated HCl by warming, and then stored in an amber coloured bottle, giving 40% stannous chloride stock solution. Just before use, 0.5ml was diluted to 66ml with DDW.

• Starch indicator Ig starch dissolved in 100ml warm (80-90°C) DDW and a few drops of formaldehyde solution were added.

• Sulfanilic acid solution 600mg sulfanilic acid dissolved in 70ml warm DDW. After addition of 20ml concentrated HCl, the volume was made upto 100ml.

• Sulphanilamide solution (1%) Ig sulphanilamide dissolved in 3N 100ml hydrochloric acid.

IV Appendix

Sulphuric acid (0.0IN) 0.272ml sulphuric acid diluted in DDW and final volume made upto 1000ml.

Sulphuric acid (0.02N) 0.544ml sulphuric acid added to DDW and the final volume made upto 1000ml.

Sulphuric acid (7N) 190.4ml concentrated sulphuric acid added to DDW and the final volume made upto 1000ml

Sulphuric acid solution 500ml concentrated H2SO4 added to 125ml DDW and cooled.