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Chaudhary Charan Singh Haryana Agricultural University, Hisar
Chaudhary Charan Singh Haryana Agricultural University popularly known as HAU, is one of Asia's biggest agricultural universities, located at Hisar in the Indian state of Haryana. It is named after India's seventh Prime Minister, Chaudhary Charan Singh. It is a leader in agricultural research in India and contributed significantly to Green Revolution and White Revolution in India in the 1960s and 70s. It has a very large campus and has several research centres throughout the state. It won the Indian Council of Agricultural Research's Award for the Best Institute in 1997. HAU was initially a campus of Punjab Agricultural University, Ludhiana. After the formation of Haryana in 1966, it became an autonomous institution on February 2, 1970 through a Presidential Ordinance, later ratified as Haryana and Punjab Agricultural Universities Act, 1970, passed by the Lok Sabha on March 29, 1970. A. L. Fletcher, the first Vice-Chancellor of the university, was instrumental in its initial growth.
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ThesisItem Open Access Biodiesel production from lipid generating microalgae(CCSHAU, 2014) Narula, Amrita; Anand, R.C.In the present scenario, the future of crude oil looks quite bleak as it may lead to all oil resources coming close to a moribund one day. Moreover, consumption of present petroleum sourced fuels at the rocket speed rate has also led to various environmental problems. One of the major concern is global warming. To make a dent in global warming, bioenergy must be generated at a very high speed. Microalgae may be the best option to produce bio-energy at rates high enough to replace a substantial fraction of fossil fuel used by our society. The primary objective of this study was to isolate lipid generating microalgal isolates for biodiesel production. The microalgal water samples were collected from ponds of different locations of eight districts of Haryana including Hisar, Rohtak, Fatehabad, Sirsa, Panipat, Karnal, Ambala and Kurukshetra. The samples were analysed for pH, EC, salinity, turbidity, total nitrogen, phosphorous and potassium. The microalgal samples were enriched in the Bold’s Basal medium and incubated at 23 ±10C, 50-55% humidity with 16:8 hours light: dark photoperiod for 21 days. A total of twenty four microalgal isolates were obtained after purification. Using Nile red staining technique microalgal isolates HMA-2 and FMA-2 were selected for further study and grown in four media of different chemical composition viz., Soil extract, Chu-13, Kuhl and Bold’s Basal medium. The microalgal isolates HMA-2 and FMA-2 produced maximum biomass (0.21 and 0.29g/l ) in Bold’s Basal medium whereas lipid content (29.6 and 25.2%) was maximum in Kuhl medium when grown at 23 ±10C, 50-55% humidity with 16:8 hours light: dark photoperiod for 21 days. Among the inorganic nitrogen sources tested potassium nitrate (original source) resulted in biomass accumulation of 0.12 and 0.19 g/l with 29.4 and 25.2% lipid content in the microalgal isolates HMA-2 and FMA-2 respectively while among organic nitrogen sources, peptone supplementation showed maximum lipid content of 38.2 and 22.1% from 0.04 g/l and 0.10 g/l biomass respectively. Similarly, among the carbon sources, glucose had been found to be a better supplement as it produced biomass of 1.59 and 1.21 g/l with 57.6 and 50.2% lipid content in microalgal isolates HMA-2 and FMA-2 respectively. Lipid and biomass production studies at different pH, temperature and salinity indicated biomass production of 0.14 g/l having 32.6% lipid at pH 8, 0.23g/l biomass having 42.4% lipid at 250C and 0.07g/l biomass containing 61.4% lipid on 20 g/l sodium chloride supplementation in microalgal isolate HMA-2 whereas in isolate FMA-2, 0.13, 0.21 and 0.11g/l biomass containing 26.1, 40.2 and 36.2% lipid was obtained under similar conditions respectively. Transesterification of algal oil of microalgal isolate HMA-2 using NaOH as catalyst and methanol resulted in biodiesel production to the tune of 9%.ThesisItem Open Access Amelioration of salt stress in chickpea (Cicer arietinum L.) by inoculation of ACC deaminase-containing mesorhizobia and rhizobacteria(CCSHAU, 2013) Chaudhary, Deepika; Sindhu, S.S.Chickpea is a major legume crop grown in the semi-arid tropics of Asia and Africa. Salinity mainly affects plant growth by decreasing the availability of water to the roots due to osmotic effect of external salts. Salinity also influences other physiological processes such as seed germination, photosynthesis, respiration and metabolite accumulation. The use of plant growth-promoting rhizobacterial as inoculants have been reported to facilitate plant growth in saline lands. Fifty isolates of Mesorhizobium were obtained from the nodules of chickpea plant samples and fifty five isolates of rhizobacteria were obtained from the chickpea rhizosphere soil collected from different part of Haryana. Among the fifty Mesorhizobium isolates, only four isolates i.e., MHD2, MSD41, MHD12 and MHD14 showed growth up to 4% NaCl salt concentration whereas 28 isolates among the 55 rhizobacterial isolates showed growth up to 4% salt with different colony size. Two Mesorhizobium isolates i.e., MHD1 and MHD12 and six rhizobacterial isolates i.e., RSD17, RSD19, RSD23, RBD12, RHD2 and RHD18 showed significant growth on ACC supplemented plates as compared to ammonium sulphate incorporated medium plates. Inoculation of selected Mesorhizobium or rhizobacterial isolates on sterilized chickpea seeds in water agar plates showed that isolate KR48 showed maximum growth of seedling roots at 5 days whereas MBD26 showed maximum growth of shoot at 10 days. Isolates RHD18, RSD3, RBD12 and RSD23 showed significant stimulation of shoot growth even at 8 dS/m salt concentration as compared to uninoculated control. Coinoculation studies with ACC+ as well as ACC- isolates of Mesorhizobium and rhizobacterial isolates were made in chickpea cultivar HC-1 under chillum jar conditions containing sloger’s broth with salt (EC, 4dS/m) and without salt. Coinoculation of ACC+ Mesorhizobium isolate MBD26 with rhizobacterial isolate RHD18 produced maximum 59 nodules/plant and 50.6% increase in plant dry weight and in the presence of salt slight decrease in nodulation (49 nodules/plant) and gain in plant dry weight (39.3%) was observed in comparison to single inoculation of MBD26 at 50 days of plant growth. At 80 days of plant growth, coinoculation of both ACC- MBD20 and RBD19 formed 41 nodules/plant and only 7.8% increase in shoot dry weight was observed as compared to the single inoculation. Coinoculation of Mesorhizobium isolate MBD26 with rhizobacterial isolate RHD18 caused increase in nodule number (78 nodules/plant), nodule weight (413 mg/plant) and shoot dry weight (61.7% increase). Whereas in the presence of salt, same treatment formed 53 nodules per plant and caused only 53.2% increase in plant biomass in comparison to single Mesorhizobium inoculated plants at 80 days of plant growth.ThesisItem Open Access Biological control of subterranean termites (Isoptera: Termitidae) with soil bacteria(CCSHAU, 2014) Dua, Seema; Sindhu, S.S.Selected ten bacterial strains having termite killing ability along with two control strains was studied for antagonistic interactions with local soil termites. Live bacterial cells, heat killed cells and culture supernatants were tested for termiticidal activity. Different bacterial strains showed more than 80% killing of termites at 5 days of incubation. Four bacterial strains KBM79, KPM35, PPM147 and PBM195 caused 100% killing at 10 days of observation. The cell free culture filtrate studies of these antagonistic cultures showed that antagonistic substance is extracellular. Termite mortality percentage increased when culture supernatants were used as compared to whole cell broth of bacterial strains. The killing frequency of the culture supernatants obtained from different bacterial strains varied from 78.0 to 96.0% at 2nd day of observation. Termicidal activity was lost on treatment of whole cell broth and culture supernatant of bacterial strains by incubation at high temperature (1000C for 10 minutes). Bacterial strains KBM79 and KPM35 possessed proteolytic, lipolytic and chitinolytic enzyme activities and caused 100% killing of termites at 10 days. Partial purified proteins caused 100% killing of termites within half an hour. The SDS–PAGE analysis of selected bacterial strains showed four common protein/polypeptides having molecular weight 129 KDa, 37.1 KDa, 28.7 KDa and 26.3 KDa in all the ten bacterial strains. These proteins/polypeptides were missing in control bacterial strains WPS73 and KPM31. Plasmid-cured strain KPM35 lost the protein band having molecular weight 129 KDa. Termiticidal activity was also decreased in plasmid-cured strain KPM35 indicating that some of the termiticidal genes were located on the plasmid. Moreover, crude preparation and partial purified protein of bacterial strains KBM79 and KPM35 when treated with proteinase K lost the antagonistic activity indicating that extracellular protein could be involved in termite mortality. Combination of different chemical compounds with efficient bacterial strains showed high termite mortality than individual insecticides. Application of bacterial strain KPM35 and dexamethasone (50 ug/ml) caused 100% mortality even at 12 hours of incubation. Dexamethasone alone was found more lethal than boric acid and sodium citrate, and caused 90% mortality at 48 hours of incubation. Bacterial strains KBM79 and KPM35 were identified as Bacillus subtilis and Pseudomonas synxantha. These bacterial strains could be further tested for termiticidal activity under field conditions.ThesisItem Open Access Simultaneous saccharification and co-fermentation of paddy straw for ethanol production(CCSHAU, 2012) Goel, Annu; Leela WatiContemporary industrial developments and rapid pace of urbanization have called for environmentally sustainable energy sources. Ethanol made from biomass can be considered as a safe and cleanest liquid fuel alternative to fossil fuels. Ethanol is economically produced from sugarcane and corn in Brazil and America, respectively. However, in a developing country like India with second largest population to feed and more than 200 million people living below poverty line sparing food crops for fuel ethanol production is not an option. Therefore, the priority in global future ethanol production is on lignocellulosic processing. Paddy straw is one of the most abundant lignocellulosic wastes on earth. Ethanol production from paddy straw is a three step process involving pretreatment, hydrolysis and fermentation. Ethanol production by separate hydrolysis and fermentation (SHF) enables enzymes to operate at higher temperature and fermenting yeasts at moderate temperatures, optimizing the utilization of sugars but to find economic acceptance, the cost for bioconversion must be lowered down and, for this, the most important process improvement made is the introduction of simultaneous saccharification and fermentation (SSF), which has further been improved to include the co-fermentation of multiple sugar substrates i.e., SSCF. Paddy straw (Pusa-1 variety) contained 35.07% cellulose, 24.85% hemicellulose, 6.29% lignin, 49.82% organic carbon and 0.85% nitrogen on dry weight basis. Alkali treatment of straw (mesh size 0.5mm) resulted in 70% lignin removal along with 88% cellulose recovery. Hydrolysis of pretreated paddy straw with commercial cellulase loaded @ 7.5 FPU/ g resulted in 75% saccharification with the release of 61% reducing sugars after 2 h of incubation at 50C. Hydrolysate supplemented with yeast nutrients (0.3% urea, 0.15% sodium dihydrogen phosphate and 0.5% yeast extract) inoculated with the co-culture of Saccharomyces cerevisiae and Pachysolen tannophilus resulted in production of 283 ml ethanol per kg delignified paddy straw at 35C after 72 h fermentation by SHF. Simultaneous accharification and co-fermentation of paddy straw supplemented with urea @ 0.3% resulted in production of 310 ml ethanol per kg delignified paddy straw with the co -culture of S. cerevisiae and P. tannophilus that was about 10 % higher than SHF. Sugar profile of hydrolysate after fermentation revealed xylose as nutilized sugar. Ethanol production efficiency in SSCF of paddy straw scaled up to 5 L capacity under optimized conditions was about 56%. The fermented residue was found to contain about 14% cellulose, 2.6% nitrogen and about 50 % residual enzyme activity. Cost of ethanol production by SSCF was lower compared to SHF in view of need of only one instrument, supplementation of cheaper yeast nutrients and without the need of buffering.ThesisItem Open Access Biological control of phalaris minor in wheat (triticum aestivum L.) using rhizosphere bacteria(CCSHAU, 2012) Phour, Manisha; Sindhu, S. S.In the present study, seventy one bacterial cultures were isolated from wheat rhizosphere soil. These bacterial isolates along with twelve reference strains were screened for their effect on seed germination of Phalaris minor on water agar plates. Ten rhizobacterial isolates i.e., HWM1, HWM7, HWM9, HWM11, HWM17, HWM30, HWM37, HWM47, HWM54 and CP43 showed maximum retardation on 5th and 10th of seed germination of Phalaris minor. At 10th day of seed germination, 14.5% bacterial isolates showed retardation of shoot growth and 19.3% bacterial isolates retarded root growth. Out of fifteen rhizobacterial isolates/strains tested for phytotoxicity effect on Phalaris minor, three rhizobacterial isolates/strains i.e., HWM11, P49 and SYB101 only caused yellowing of leaves whereas eight rhizobacterial isolates/strains HWM10, HWM25, KPM15, SB153, PPM126, WPS73, CPA152 and GYB106 caused appearance of disease spot. Screening of rhizobacterial isolates for production of indole acetic acid showed that nine isolates i.e., HWM7, HWM11, HWM18, HWM23, HWM24, HWM37, HWM42, HWM57 and PPM115 produced IAA ranging from 7.0-10.0 Og/ml. Two isolates HWM49 and HWM35 produced 11.10 and 14.07 Og/ml IAA, respectively. Maximum production of IAA (> than 25 Og/ml) was observed in isolates CPS67, CP43 and HWM13. Rhizobacterial isolates/strains HWM59, HWM69, CP43 and CPS67 showed >7.0 Og/ml production of δ-aminolevulinic acid. Other 22 bacterial isolates produced ALA ranging from 2 to 7 Og/ml. Fungal growth inhibition studies showed that 20.5% of total rhizobacterial isolates inhibited the growth of Fusarium oxysporum on PDA medium plates. Large zone of inhibition was formed by bacterial isolates HWM13, WPS73, SB153, HWM25, HWM31, HWM37 and HWM57. Twelve rhizobacterial isolates/strains were tested for their effect on growth of wheat and weed under pot house conditions. Rhizobacterial strains/isolates i.e., SYB101, CPS67 and HWM11 were found to stimulate growth of wheat and inhibited the growth of Phalaris minor.ThesisItem Open Access Simultaneous saccharification and fermentation of sugarcane bagasse to ethanol(CCSHAU, 2013) Meenakshi; Leela WatiThe use of sugarcane bagasse in bioethanol production provides an alternative opportunity for more sustainable development of renewable resources. Sugarcane bagasse contains 30-40% cellulose, 20-25% hemicellulose and 18-24% lignin. Glucose and xylose sugars can be derived from the cellulose and hemicellulose portion of sugarcane bagasse that can be fermented into ethanol. Ethanol can be produced from sugarcane bagasse using three main steps: pretreatment; hydrolysis and fermentation. Pretreatment must be cost-effective besides improving the formation of sugars by hydrolysis. Alkali (Sodium hydroxide and lime) pretreatment of lignocellulosic materials reduce the crystallinity of cellulose and increase the porosity of the lignocellulosic materials. Pretreated bagasse can be fermented either by separate hydrolysis and fermentation (SHF) or simultaneous saccharification and fermentation (SSF). Simultaneous Saccharification and Fermentation improves reducing sugar availability as sugars released by enzymatic activity are used imultaneously and prevent product inhibition by glucose. Other advantage is production of more ethanol in less time because it reduces one step as hydrolysis and fermentation are carried out only in one reactor. For the present study sugarcane bagasse collected from sugar mill, Meham (Rohtak) was ground to 2.0 mm (coarse) and 0.5 mm (fine) particle size. Sugarcane bagasse had 26.5% cellulose, 20.8% hemicellulose and 18.3% lignin content before treatment. Sugarcane bagasse of three particle sizes (unscreened and screened to 2.0 and 0.5 mm) was pretreated with 1 and 5% NaOH with boiling, autoclaving and microwaving treatments and also pretreated with lime. Maximum 81.42% reduction in lignin content was obtained on treatment of coarsely ground bagasse (2.0 mm particle size) boiled in 5% NaOH for 60 min. Solid recovery after various pretreatments to different particle size sugarcane bagasse varied between 40 to 78% and a maximum of 82.70% cellulose and 45.34% hemicellulose was recovered from coarsely ground bagasse boiled in 5% NaOH for 60 min which was used for fuel ethanol production by simultaneous saccharification and fermentation due to high efficiency of delignification. Among different commercial enzymes studied, palkonol, had highest Exoglucanase (13.53 U/ml), endoglucanase (26.67 U/ml) and xylanase (16.46 U/ml) activity. Enzymatic saccharification of pretreated bagaasse with Palkonol loaded at 2% (v/v) released maximum (45.2 g /100g) total reducing sugars after 2 h reaction at 50°C. Simultaneous saccharification and fermentation (SSF) of pretreated sugarcane bagasse (10% w/v) using yeast Candida utilis (1% w/v) with 2% v/v enzyme loading and supplemented with 0.3% urea resulted in production of 2.2% (v/v) ethanol after 72 h fermentation at 35°C. On other hand, separate hydrolysis and fermentation (SHF) of sugarcane bagasse carried out under similar conditions, however resulted in lesser (0.9% v/v) ethanol production. Simultaneous saccharification and fermentation of coarsely and finely ground bagasse resulted in production of similar concentration of ethanol i.e. 2% (v/v) under optimized conditions.ThesisItem Open Access Bacterial community structural analysis of saline and sodic water irrigated soils and its effect on wheat growth(CCSHAU, 2014) Harshpreet, Kaur; Gera, RajeshThe problem of the salinity or sodicity existed long before the start of agricultural practices. It has now become a very serious problem for crop production, particularly in arid and semi-arid regions, which constitute about one third of the world’s land surface. Irrigation water always includes some amount of dissolved substances, collectively called salts. Due to inadequate and unassured supplies of good quality water in many arid and semi-arid regions of the world, farmers use saline or sodic ground water for irrigation of crops. Long-term and indiscriminate use of such water causes accumulation of salts, particularly Na, which not only adversely affects the physical and chemical properties of soil, but also modifies the quality of soil organic carbon and its biotic components. Saline water is categorized as water with an EC > 4.00 deciSiemens / meter (dS/m) while sodic water is categorized as water with an RSC > 2.5me/l. The effects of salinity or sodicity on soil chemical and physical properties are well known and their effects on the soil microbial community and microbiological processes remain relatively unstudied. Thus keeping in view the above facts, a total of 15 saline water irrigated rhizospheric soil along with their respective water samples were collected from Hisar, Jind, Sirsa and Rewari districts. The pH and ECiw of these water samples varied from 6.00 - 8.20 and 2.48 - 17.14 dS/m respectively. Their respective soil samples were analyzed for pH, ECe, organic C, total N and total P, which varied from 6.04 - 8.10, 4.10 - 21.00 dS/m, 0.13 - 0.45%, 0.012 - 0.044 % and 587 - 703 mg P/kg soil, respectively. Similarly, a total of 20 sodic water irrigated rhizospheric soil and their respective water samples were also collected from Hisar, Mahendergarh, Karnal and Kaithal districts. The water samples were analyzed for the RSC, pH and ECiw, which was above 2.50 me/l, 8.50 - 10.90 and 1.01 - 1.50 dS/m, respectively. The pH, ECe, organic C, total N and total P of soil samples ranged from 8.30 - 10.50, 1.00 - 2.40 dS/m, 0.09 - 0.46%, 0.002 - 0.040% and 625 - 701 mg P/kg soil, respectively. The viable counts of bacteria for the above soil samples collected from saline and sodic soils were taken on Jensen’s, malate, Pikovaskaya, King’s B media and soil extract agar. It was observed that with the increase in EC, there was decrease in viable counts. A total of 215 morphotypes were obtained from the above 35 different soil samples, out of which 91 and 124 morphotypes belonged to saline and sodic soils, respectively. Most of these morphotypes were identified as gram -ve rods, cocci and small rods in addition to some fraction of gram +ve small rods and cocci. All these morphotypes were characterized for agriculturally useful traits such as P-solublization, siderophore production, IAA production and ammonia excretion. Out of 91 morphotypes obtained from saline and 124 morphotypes from sodic soils, only 15 and 18 morphotypes, respectively, were found to have all the above four characteristics. In saline soil, more numbers of morphotypes were ammonia excreters while in sodic soil, IAA producers were present more in numbers.The genomic DNA of the morphotypes obtained on soil extract, Jensen’s and malate media was isolated by the CTAB method and was amplified for nifH gene by using degenerative primers, nifH for and nifH rev. Out of 57 morphotypes obtained from saline soil and 86 morphotypes obtained from sodic soil, 28 and 42 morphotypes, respectively, showed nifH gene amplification, indicating that these morphotypss can fix nitrogen. The efficiency of six promising bacterial morphotypes isolated each from saline (JK246, JJ355, SK543, JS 509, RS 502 and JP323) and sodic soils (KtJ571, KtK569, KrK564, KrS530, KrS546 andKtP390) were checked in wheat variety KRL 210 at different levels of induced salinity having an EC of 0, 4, 8 and 12 dS/m and sodicity with RSC 0, 10 and 15 me/l, respectively. The bacterial counts were less in uninoculated plants as compared to inoculated one. Moreover, in all the treatments, the total bacterial counts were higher at 45th day of sowing then the counts recorded before sowing and as the crop reached to maturity, the viable counts start decreasing. Best results were observed at EC 4 dS/m and RSC 10 me/l in terms of plant height, no. of tillers, plant dry weight and mean grain weight. The significant difference with respect to these parameters was observed in all the induced EC or RSC levels. The inoculation of different morphotypes isolated from saline or sodic soils resulted in a significant increase in plant growth parameters as compared to control or uninoculated plants. In all the treatments with different EC or RSC levels, the consortia performed better as compared to individual morphotypes and the reference strains i.e. Mac 27 and P-36 in terms of plant growth parameters both in saline (SK543+ JS 509+ RS 502+ JP323) as well as sodic (KrK564+ KrS530+ KrS546+ KtP390) soils.ThesisItem Open Access Role of ACC (1-aminocyclopropane-1-carboxylate) utilizing pseudomonas and mesorhizobium in disease control and plant growth stimulation of chickpea (Cicer arietinum L.)(CCSHAU, 2012) Kiran Kumari; Sindhu, S. S.Chickpea is an important commercially utilizable crop and its high yielding varieties are more susceptibile to soil borne fungal phytopathogens i.e., Fusarium oxysporumand Rhizoctonia bataticola. Recently, ACC deaminase containing Mesorhizobium and Pseudomonasbacteria have been found to improve nodulation and suppressed disease in chickpea by lowering the level of stress ethylene. In this, study, 85 Mesorhizobiumisolates and sixty one Pseudomonas isolates were screened for utilization of ACC on Dworkin and Foster’s minimal medium. Frequency of Mesorhizobium and Pseudomonas isolates having the ability to utilize ACC was found 30.6 and 14.3%, respectively. Among ACC + isolates, 7.37% isolates ofMesorhizobiumshowed antifungal activity against F. oxysporumand 6.32% isolates inhibited the growth of R. bataticola.on PDA plates. Screening of Pseudomonas isolates showed that 21.98 and 47.5% isolates inhibited the growth of F. oxysporum and R. bataticola, respectively. Mutant altered in ACC utilization were derived from Mesorhizobium andPseudomonas isolates by using nitrosoguanidine. Seven mutants fromPseudomonasisolate PPBM36 and 5 mutants from Mesorhizobiumisolate CMK39 lacking ACC utilization were obtained. Mesorhizobium isolates i.e., CMK12, CMK39, CRYM13 and CMK21 formed 75 to 98 nodules per plant and 229.3 - 278.5% increase in plant dry weight was observed at 90 days of growth in chillum jar assembly conditions. Coinoculation of Mesorhizobium isolate, CMK39 with Pseudomonas isolate PPBM36 formed 73 nodules per plant and caused 48.4 - 127.5% increase in root and shoot dry weight at 90 days ofgrowth. In Fusarium oxysporuminoculated soil, single inoculation of Mesorhizobiumisolate CMK39 caused 46.5% increase whereas its coinoculation with Pseudomonasisolate PPBM36 further enhanced shoot dry weight upto 67.5% in comparison to uninoculatedcontrol. Disease caused byF. oxysporum was effectively controlled by coinoculation of Mesorhizobium isolate CMK39 with Pseudomonas isolate PPBM36. ACC utilizing Mesorhizobium and Pseudomonas isolates were found more effective in control of disease symptoms as compared to ACC - isolates and mutants. Thus, ACC utilizing Mesorhizobium and Pseudomonas isolates could be used for disease control and plant growth promotion of chickpea under field conditions.ThesisItem Open Access Evaluation of mycobiota of spoilt wheat in expediting ethanol production(CCSHAU, 2014) Preeti; Goel, SnehEthanol besides being a known feedstock for chemical and beverage industry is also being used as an additive to fuel for automobiles. Over the years, its demand in general, has been on an increase and India is no exception. Clearly, maximization of ethanol production becomes imperative and could be achieved by considering, among others, the addition of yeast nutrients to the fermentation liquor. These nutrients by accelerating the rate of fermentation reduce the fermentation time and in turn lead to maximization of ethanol volumes on a daily basis. Usually, N and P represent these nutrients and in the Indian distilleries they are conventionally sourced from chemical fertilizers, which do have a certain carbon footprint. Ideally, organic/ renewable yeast nutrients would be more suitable, as they have a low net green house gas emission. However, so far, no dedicated nutrient formulation is available in the Indian market. Spoilt wheat which ferments faster than its normal version has been shown to offer a potential for such a formulation, as it contains factor(s)/activity that is responsible for expediting rate of ethanolic fermentation. This ethanol-expediting activity has been speculated due to in situ production of yeast nutrients viz., low molecular weight peptides and free amino acids by the activities of resident bacterial and fungal flora of spoilt wheat. So, fungal flora of spoilt wheat was screened for its ethanolexpediting capability, leading to the retrieval of the best isolate. Such an isolate might later contribute to the development of more useful, environment friendly and activity-enriched wheat-based supplements to expedite ethanolic fermentation by yeast. The spoilt wheat (SW) sample tested positive for ethanol-expediting activity on 30% normal wheat (NW) hydrolysate, as its supplementation @ 15%, both as coarse flour or its aqueous extract, produced 10.6% and 11% (v/v) ethanol, respectively, against 8% (v/v) by the control. Clearly, the SW sample under study was fit for isolation of fungi. Based on colony morphology, 30 fungal isolates were recovered from the SW sample. Screening of the isolates for amylolytic, proteolytic and lipolytic activities on the plate revealed that all the 30 isolates were positive for the amylolytic, 13 were positive for proteolytic and 18 for the lipolytic activities. Thus, only three bacterial isolates viz., SWF-6, SWF- 11 and SWF-20 having dissolution factor of >1 were further utilized to explore their ethanol-expediting capabilities by producing laboratory spoilt wheat grains and subsequently estimating ethanol production by yeast on 30% laboratory developed spoilt wheat as well as on 30% normal wheat (NW) hydrolysate, as its supplementation @ 15%, both as coarse flour or its aqueous extract. Ethanol estimation at 24h showed that three isolate viz., SWF-6, SWF-11 and SWF-20 did produce a boost to alcohol production from 11 to 12.2% (v/v). These three isolates based on morphological characterization were identified to belong most probably to genus Aspergillus.
