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Dr. Y. S. Parmar University of Horticulture & Forestry, Solan
Dr. Yashwant Singh Parmar University of Horticulture and Forestry, Solan, was established on 1st December, 1985 with the objective to promote education, research and extension education in the fields of Horticulture, Forestry and allied disciplines. Late Dr. Yashwant Singh Parmar, the first Chief Minister and the architect of Himachal Pradesh perceived the importance of Horticulture and Forestry to develop and improve the State economy which led to the establishment of this University. Its history lies in erstwhile Himachal Agricultural College, Solan, established in 1962 and affiliated to the Panjab University. It became one of the campuses of Agriculture Complex of Himachal Pradesh University on its formation in 1970. Consequent upon the establishment of Himachal Pradesh Krishi Vishvavidyalaya in 1978, this campus became its Horticulture Complex and finally in 1985, assumed the status of a State University, being the only University in the country engaged exclusively in teaching, research and extension in Horticulture and Forestry.
The University is located at Nauni in Solan District of Himachal Pradesh, 13 km from Solan on Solan-Rajgarh Road, at an elevation of 1300 metres above mean sea level. Solan town is situated on national highway (NH-22) and is well connected by train and bus services.
The University has four constituent colleges, out of which, two are located at the main campus Nauni, one for horticulture and the other for forestry, having 9 and 7 departments, respectively. The third College i.e., College of Horticulture & Forestry is located at Neri in Hamirpur District on Nadaun-Hamirpur state highway, about 6 Km from Hamirpur town and is well connected with bus service. The college offers three Undergraduate Degree Programmes i.e. BSc (Hons.) Horticulture, BSc (Hons.) Forestry and B. Tech. Biotechnology and MSc degree programme in a few subjects. The fourth college i.e. College of Horticulture and Forestry, Thunag (Mandi) is located at Thunag District Mandi. This college offer BSc (Hons.) Horticulture and BSc (Hons.) Forestry degree programme. In addition, there are five Regional Research Stations, 12 Satellite Stations and five Krishi Vigyan Kendras (KVKs) situated in different zones of the State.
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ThesisItem Open Access AN INNOVATIVE GREEN TECHNOLOGY FOR AGRICULTURAL WASTE UTILIZATION(NAUNI,UHF, 2020-10) SHARMA, KANIKA; SHARMA, NIVEDITAABSTRACT In the present investigation, different agricultural wastes were collected from Himachal Pradesh and their adjoining states i.e. Punjab and Haryana and an attempt had been made to utilize this waste as substrate for its enzymatic degradation to enhance their saccharification and bioconversion to ethanol by co-fermentation with ethanologens. Based upon the chemical composition of mixed agricultural waste and large availability of rice straw, these were finally selected for further study. Different physico-chemical pretreatment methods applied to rice straw/mixed waste, where maximum reducing sugars i.e. 46.98 and 44.45 mg/g from rice straw/mixed waste respectively were obtained in microwave pretreatment. After optimization of different process parameters by one factor at a time (OFAT) approach for enzymatic saccharification under SmF, highest reducing sugars reported from untreated and pretreated rice straw/mixed waste were 34.40 and 56.70 mg/g; 38.0 and 63.50 mg/grespectively, at enzymatic ratio of 3.5:1.5, 50oC temperature, 5.5 pH, incubation period 72 h and 7.5% biomass loading followed by CCD of RSM by varying temperature, pH and incubation time. SEM analysis revealed that the surface structure of rice straw/mixed waste was significantly changed after pretreatment and enzymatic hydrolysis. The reducing sugars so obtained were analyzed using HPLC technique. In 2nd mode i.e. SSF (biological degradation), two fungal strains selected P. chrysoporium and P. ostreatus fungal strains had showed maximum amount of reducing sugars produced i.e. 55.70 and 49.05 mg/g by using microwave pretreated rice straw and mixed waste respectively. After optimization of process parameters through OFAT and RSM maximum reducing sugars from untreated and pretreated rice straw (450 watt) /mixed waste (600 watt) had the critical values as 51.12 and 37.98 mg/g; 71.99 and 73.10 mg/g respectively, at 30 and 25oC temperature and incubation period 10 days respectively. Among two processes of ethanol fermentation evaluated in the present study, SHF was found to be the best in terms of highest ethanol productivity with S. cerevisiae I+ P. stipitis as best fermenting microorganism from enzymatic saccharified and fungal degraded sugary syrup. Estimation of bioethanol production after enzymatic saccharfied/ biological degrdaded microwave pretreated rice straw/mixed waste hydrolysate fermented by co-culture of S. cerevisae I and P. stipitis was done with the help of GC-MS. Scale up of ethanol fermentation using 7.5 l stirred tank bioreactor, batch conversion of rice straw/mixed waste enzymatic saccharified hydrolysate to ethanol was carried out by co-culture of S. cerevisiae I + P. stipitis under SHF. The highest ethanol yield of 52.14 and 46.89 g/l with 88.12 and 80.01 % of fermentation efficiency was achieved after 48 and 36 h of fermentation with an agitation speed of 200 rpm and aeration rate of 0.05vv, pH 6 at 25 ±2°C and inoculating the bioreactor with 24 h old co-culture of yeasts (S. cerevisiae I + P. stipitis). Also to combat the pollution problem, value addition of agricultural waste for cultivation of mushroom supplemented with apple pomace had emerged as an ecofriendly technique with higher yield and better utilization substrate.ThesisItem Open Access STUDIES ON THE EFFECT OF INOCULATED Pseudomonas STRAINS ON PLANT GROWTH PROMOTING POTENTIAL OF RHIZOSPHERIC SOIL OF APPLE(UHF,NAUNI, 2016) POOJA, DEVI; KAUR, MOHINDERABSTRACT Apple replant disease has plagued apple growers for many years and is now found in most fruit growing regions world-wide. The use of plant growth promoting rhizobacteria (PGPR) for the benefits of agriculture and horticulture is gaining worldwide importance and viewed as a novel and potential tool for providing substantial benefits to the horticulture. In the present study, isolation and characterization of fluorescent Pseudomonas sp. from replanted site of apple orchard (Shimla HP) was done. The eleven Pseudomonas isolates and four already isolated Pseudomonas strains L, M, C and G were screened out for various plant growth promoting activities like siderophore, phosphate solubilization, antifungal activity, plant growth regulators (auxins, cytokinins and gibberellins), lytic enzymes and production of HCN and ammonia. On the basis of PGPR activities, two isolates (ArSh-3 and ArSh-4) were genotypically characterized by 16S rRNA gene sequencing. Already isolated Pseudomonas strains L, M, C and G used in different liquid formulations of individual and consortium fluorescent Pseudomonas to treat apple plants in replanted site of orchards at Sharontha and Siao. The performance of apple plants was much better in terms of root colonization capacity, plant establishment and increase in plant growth in terms of plant height, number of nodes, chlorophyll content of leaves, plant growth promoting activities and NPK of rhizosphere soil over their respective control after twelve months of treatment. These strains can be further exploited for management of replant problem of apple after conducting few more field trials in replanted sites and can have great importance in the field of horticulture.ThesisItem Open Access BIOFUEL PRODUCTION FROM APPLE POMACE- A MAJOR HORTICULTURAL WASTE OF HIMACHAL PRADESH(UHF, NAUNI, 2016) PATHANIA, SHRUTI; SHARMA, NIVEDITAABSTRACT In the present investigation, an attempt was made to utilize apple pomace as substrate for its degradation by potential microorganisms and evaluated process parameters to enhance their rate of hydrolysis a key step for its bioconversion to ethanol by co-fermentation with immobilized yeast cell. In total 7 fungi were isolated, among them, F2 was screened for multiple carbohydrases production and was identified using ITS 5.8S rRNA technique as R. delemar F2 and registered under accession no. KX5123312. Multiple carbohydrases production was optimized through classical approach one factor at a time (OFAT) varying microwave dose, moisture, temperature and incubation time. The partial purification of multiple carbohydrases was done by cold acetone precipitation and carbohydrases from bacteria by ammonium sulphate precipitation. To reduce the production cost of ethanol, cheap untreated and pretreated cellulosic biomass i.e. apple pomace used as a substrate for sugar production. Bioconversion of microwave pretreated apple pomace to ethanol was studied under two different fermentation processes i.e separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF). The optimum parameters obtained for the saccharification of microwave pretreated apple pomace using carbohydrases from bacteria were at pH 5.0, 10% (w/v) of biomass loading,500C at 6:1:2:1 of enzymatic loading after 36 h of incubation with fermentable sugar yield of 36mg/g of reducing sugar and 62.01mg/g of total sugar. Similarly sacchaification of 450 watt dose of microwave pretreated apple pomace with multiple carbohydrases from R.delemar F2 yielded 20.87mg/g of reducing sugar and 28.36mg/g of total sugar at enzymatic loading of 20ml/5gm at 500C after incubation for 46h under SHF. Structural changes of apple pomace before and after enzymatic pretreatment were further investigated through ccanning electron microscopy (SEM), monosaccharide content in the saccharified samples was quantified using high performance liquid chromatography (HPLC) system. Maximum ethanol yield of 48.19 g/l with fermentation efficiency of 61.42% was achieved by immobilized co-culture of S. cerevisiae I + P. stipitis in sodium alginate with 4.5 mm of bead size from hydrolyzed syrup of apple pomace from bacterial carbohydrases. Scale up of SHF with apple pomace using carbohydrases from bacteria and fungus with immobilized co-culture of S. cerevisiae I + P. stipitis was performed in 7.5 L bioreactor, achieving highest ethanol production after 36 h of fermentation with 36.97g/l of ethanol (72.35% of fermentation efficiency) and 31.60g/l of ethanol yield (with 61.80% of fermentation efficiency) at 100 rpm of agitation rate, 0.05vvm of aeration rate respectively, observed the best combination for highest bioethanol production. Also the value addition to the left over residue of apple pomace for single cell protein production and mushroom production help to dispose this waste in an environment friendly manner and production of microbial protein.ThesisItem Open Access IMPACT OF INDIGENOUS PLANT GROWTH PROMOTING RHIZOBACTERIA AND CHEMICAL FERTILIZERS ON SOIL HEALTH AND PRODUCTIVITY OF CAPSICUM (Capsicum annuum L.)(UHF,NAUNI, 2016) GUPTA, SHWETA; KAUSHAL, RAJESHABSTRACT Capsicum is a renumerative crop to the farmers and is grown in both open fields and protective structures. The nutrient requirements and its sensitivity to extremes of environment coupled with higher incidence of diseases such as damping off (Pythium spp.), phytophthora collar rot (Phytophthora capsici), fruit rot (Colletotrichum capsici) and bacterial wilt (Ralstonia solanacearum) resulted in sizeable yield losses. The use of PGPR to supplement chemical fertilizers and pesticides is a potential alternative but no commercial formulation is available for use. So, the present investigations entitled âImpact of indigenous plant growth promoting rhizobacteria and chemical fertilizers on soil health and productivity of capsicum (Capsicum annuum L.)â was carried out during 2013-2016. Rhizosphere and root samples of capsicum were collected from twenty eight locations of agro-climatic zone I, II and III of Himachal Pradesh. Among 157 isolates, 34 morphological distinct isolates were selected for screening of possession of multifarious plant growth promoting traits. Among selected isolates, all were P-solubilizers, nitrogen fixers, siderophore producers, 18 isolates were able to produce IAA, 19 isolates were ACC- deaminase producers, 12 isolates were ammonia producers and only 5 isolates were HCN producer. Under laboratory conditions, except THE 17 isolate, all the tested isolates were able to inhibit the growth of one or more test pathogens i.e. Fusarium solani, Rhizoctonia solani, Pythium spp., Ralstonia solanacearum, Phytophthora capsici and Colletotrichum capsici. Fifteen isolates possessing maximum PGP traits were characterized further. The TCP solubilisation by selected fifteen isolates ranged from 169.84 ”g/ml to 60.16 ”g/ml, siderophore production efficiency ranged from 56.36 percent to 12.79 per cent, able to grow in a temperature range of (20 - 40ÂșC), pH (5.0-8.0) and salt concentration (2-6%). On the basis of 16S rRNA gene sequencing three isolates (JHA6, ROH6 and ROH14) possessing maximum multifarious PGP traits were identified as Providencia sp. (ROH6), Pseudomonas aeruginosa (JHA6) and Bacillus amyloliquefaciens (ROH14). The application of isolated indigenous PGPR Providencia sp. (ROH6) reduced the disease incidence of bacterial wilt (Ralstonia solanacearum) by 70% as compared to pathogen inoculated control. The conjoint application of PGPR isolate (JHA6 and ROH14) along with 80 per cent NP (N80 and P61 kg/ha) brought a significant increase in yield by 8.93%, available N and P contents by 8.64 and 20.73 per cent, over recommended doses (N100 and P76 kg/ha) besides saving 20 per cent chemical fertilizers. The study, therefore, indicates the potential of Bacillus amyloliquefaciens (ROH14) and Pseudomonas aeruginosa (JHA6) in partial replacement of N and P (~20%) applied through chemical fertilizers, besides higher productivity of crops. Further, the application of selected indigenous isolates (JHA6 and ROH14) can induce drought tolerance by increasing antioxidant enzymes (superoxide dismutase, catalase and peroxidase) activities. Thus, the conjoint application of PGPR isolates at 80 per cent doses of N & P has good prospects to be used as biofertilizer, biocontrol and biostimulant not only for enhanced growth and yield of capsicum but also to sustain soil health.ThesisItem Open Access PRODUCTION AND PURIFICATION OF PECTINASE AND ITS APPLICATION IN PREPARATION OF PROBIOTIC FORTIFIED FUNCTIONAL FRUIT JUICES(UHF,NAUNI, 2016) HANDA, SHWETA; SHARMA, NIVEDITAABSTRACT Fruit compost being a highly probable source for pectinolytic microorganisms was utilized as a source for isolation of pectinolytic microorganisms. In total, 16 pectinolytic bacteria and 8 fungi have been screened from fruit compost. Among them, two bacterial strains C1 and S5 and one fungal strain C4 were selected on the basis of highest pectinase production (IU/ml) and were identified as B. parabrevis C1 |KU323596|, S. violaceoruber S5 |KX512313 | and R. sexualis C4 |KX512311|. Cultural conditions and process parameters viz. media types, pH, temperature, inoculum size, incubation time, substrate concentration, divalent ions and surfactants etc. were optimized firstly through classical one variable at a time (OVAT) followed by statistical optimization by employing central composite design of response surface methodology. The enzymes obtained from all the three strains were purified to homogeneity by following a sequential purification approach. B. parabrevis C1, S. violaceoruber S5 and R. sexualis C4 pectinase was purified to a final purification fold of 2.26, 3.31 and 2.68 respectively. Molecular weight of B. parabrevis C1 and R. sexualis C4 pectinase was 43 kDa whereas; S. violaceoruber S5 pectinase had a molecular weight of 60 kDa. Pectinase activity was found to be maximum at 50 oC and pH 6.0 for B. parabrevis C1 and R. sexualis C4 and 60 oC and pH 9.0 for S. violaceoruber S5. Pectinase from all the three strains was quite thermostable with retention of more than 50% activity after incubation of 90 min at 45-60 oC. Kinetic characteristics of pectinase from all the three strains showed that the enzyme was very efficient qualitatively as well as quantitatively. Raw pectin hydrolysis ability shown by pectinase from all the three strains is a rare feature of pectinase making it a potential candidate for industries. Purified pectinase of S. violaeceoruber S5 being most potential among all the pectinases study along with the other hydrolytic enzymes viz. cellulase and amylase was employed for the juice extraction and clarification by statistical optimization by employing central composite design of response surface methodology. Treatment 21 having pectinase 10U, Cellulase 2U, Amylase 5U at temperature 52 oC in time 60 min showed maximum apple juice yield and clarification. Further, apple juice was fermented with the inhouse probiotic strains i.e. L. reuteri F8, L. delbreuckii subsp. bulgaricus and L. plantarum F22 in different sets, and Set D having consortia of these three probiotic strains was highly nutritious having high content of ascorbic acid, phenols and proteins and was successfully accepted in its sensory evaluation. The apple juice proved to be a suitable media for the production of a fermented probiotic drink and can serve as a healthy beverage.ThesisItem Open Access STUDIES ON CHARACTERIZATION AND BIOCONTROL POTENTIAL OF CHITINOLYTIC PLANT GROWTH PROMOTING RHIZOBACTERIA(2016) THAKUR, DEEPALI; CHAUHAN, ANJALIABSTRACT Plant growth promoting rhizobacteria (PGPR) are root associated bacteria and benefit plants by providing growth promotion through a variety of mechanism. The public concern over the harmful effects of chemical pesticides on the environment and human health has enhanced the search for safer, environmentally friendly control alternatives. Due to the importance of chitinolytic enzymes in insect, nematode, and fungal growth and development, they are receiving attention in regard to their use as biopesticides and microbial biocontrol agents. Therefore, the present study was aimed to characterize rhisospheric bacterial isolates associated with various horticultural crops and medicinal plants. A total of eighty two bacterial isolates were screened for chitinase production on minimal agar medium where zone of clearance and enzyme activity index were used for initial screening of chitinolytic bacteria. Fifty two out of eighty two bacterial isolates were selected as chitinase producers on the basis of zone size ranging from 1.60 mm to 45.40 mm and were further screened for quantitative chitinase production. All the fifty two isolates were screened for multifarious plant growth promoting traits viz. phosphate solubilisation, IAA production, siderophore production, HCN production and antifungal activity against Fusarium oxysporum and Phytopathora capsici. Five bacterial isolates which exhibited maximum chitinase activity along with maximum plant growth promoting traits were further evaluated for their biocontrol potential against Fusarium wilt of tomato caused by FOL under net house conditions. One isolates CT59T showed maximum biocontrol efficacy of 60.34 % against Fusarium wilt in tomato under net house studies comparison to control where disease index of 91.67 % was recorded. On the whole maximum disease control was found in case of fungal challenged tomato plant initially treated with CT59T. The identification of CT59T was tentatively confirmed by biochemical characterization and it was identified as strain belonging to Bacillus sp. The study on chitinase production by bacterial isolate was optimized under different cultural conditions resulted in maximum chitinase production (52.80 U/ml) after 72 h of incubation at 40 °C in minimal salt medium of pH 5.0 with an inoculum size of culture of 5.0 %. Bacterial isolate CT59T was grown at optimum culture conditions and chitinase was partially purified by ammonium sulphate precipitaion at 20-70% concentration and by dialysis of the proteins. The partially purified enzyme was characterized for stability of enzyme activity and it was found to be stable within a pH range of 5-7, temperature range of 30-40 ÂșC and reaction time of 90-120 minutes. The inhibition effect of crude and partially purified chitinase enzyme activity from Bacillus sp. strain CT59T on Fusarium oxysporum f.sp. lycopersici showed its potential as biocontrol agent.ThesisItem Open Access CHARACTERIZATION OF BIOACTIVE METABOLITES PRODUCING STRAINS OF Pseudomonas SPECIES FOR SURVIVAL AND GROWTH PROMOTION OF REPLANTED APPLE(2016) KAUNDAL, KIRTI; KAUR, MOHINDERABSTRACT The present study focuses on characterization of secondary antifungal metabolites i.e. Phenazine and Pyrrolnitrin produced by fluorescent Pseudomonas sp. isolated from the normal and replant sites of Maggota and Sioa of apple orchards. Twelve strains were isolated from the replant sites of Maggota and Siao of Shimla distt. and four already isolated strains of fluorescent Pseudomonas sp. from replant sites of apple were screened out for direct and indirect plant growth promoting activities like antifungal, siderophores, phosphate solubilization and production of HCN, ammonia, plant growth regulators (auxins, cytokinins and gibberellins) and lytic enzymes. By 16S Rrna gene characterization isolates An-3-Mg and An-19-Mg showed 99% similarity with Pseudomonas poae and Pseudomonasputida respectively. Three fungal isolates from replant site of orchard at Maggota showed similarity i.e. ARF1 (Fusarium oxysporum, 99%), ARF2 (Acremonium zeae, 97%) and ARF3 (Curvularia australiensis, 99%) and one isolate from replant site of orchard at Siao i.e. ARF4 99% similarity with Plectosphaerella cucumerina by ITS gene sequencing method. Optimization of media, time of incubation, temperature, pH, carbon and mineral sources were done for production and extraction of antifungal metabolites i.e. Phenazine & Pyrrolnitrin and for mass multiplication of inoculums for field trials. These metabolites were extracted with benzene and acetone identified and characterized through TLC at Rf 0.52 and 0.25 respectively. Antifungal activity, MIC and thermal stability of both these metabolites were evaluated. Phenazine was further characterized through HPLC, NMR and FTIR techniques. The extracted metabolite showed inhibition of fungal pathogens i.e. 24.2 to 44.2%. Individual and consortial formulation of four best selected strains i.e. P. putida (L), P.fluorescens(M) and P.aeruginosa (G and H) showed growth promotion in terms of plant height, no. of nodes, Chl content, available NPK and available micronutrients of replanted apple at Maggota and Siao. Therefore these strains can be exploited for the management of replant problem of apple. SignatureThesisItem Open Access STUDY OF RHIZOBACTERIAL COMMUNITY STRUCTURE AND EVALUATION OF PLANT GROWTH PROMOTING RHIZOBACTERIAL APPLICATION STRATEGIES TO PROMOTE GROWTH AND YIELD IN APPLE(2016) CHANDEL, SHALINI; SHIRKOT, C.K.ABSTRACT Apple (Malus domestica Borkh.) is one of the most important fruit crops grown and consumed in the world. In Himachal Pradesh, apple crop occupies an area of 1,07,700 hectare with a production of 7,38,700 metric tons. Attempts to enhance apple productivity, in the Trans Himalayan region has resulted in rapid expansion in area under apple cultivation. However, the strict physio-geographic conditions, pose several hurdles. High yielding modern varieties are being adopted, to increase the productivity, which are highly responsive to external inputs. This results in indiscriminate use of chemical inputs, which poses potential threat to human health and environment. The utilization of plant growth promoting rhizobacteria (PGPR) may help to develop an environmentally benign biological approach for managing fungal diseases and enhancing the plant health resulting in increase in yield. In the last decade of research, several PGPR strains that have been identified and the research on PGPR has seen a tremendous boost, mainly because the role of the rhizosphere as an ecological unit has gained importance in the functioning of the biosphere. The present study envisages better understanding of structural and functional rhizobacterial diversity associated with apple replant sites and root rot infected diseased apple orchards. Principal component analysis revealed detailed insight into correlation between apple associated rhizobacterial population and different plant growth promoting traits in (a) healthy and root rot infected apple orchards; (b) apple replant sites and new plantation apple orchards. Molecular diversity analysis employing techniques viz. amplified ribosomal DNA restriction analysis (ARDRA) and DNA sequence analysis of 16S rRNA gene revealed comparative insight into structural variations in rhizobacterial diversity. In planta evaluations using microbial formulations developed using PGPR strains viz. Bacillus licheniformis strain CKA1, B.methylotropicus strain CKAM, Serratia plymuthica strain G4(3), B.methylotropicus Strain RG1(3), and their consortium have been comprehensively evaluated in the present study. Bio-control potential of microbial formulations against phyto-pathogens has also been assessed using in planta experimentation. Strain CKA1 based bio-formulation proved to be the most effective as compared to all other PGPR strains in providing antagonism against pytopathogens as revealed by experimentations carried out under net house conditions. Attempts have also been made, in the present study to understand the genetic basis of mineral phosphate solubilization in strain CKA1. Cloning and sequencing of gdh gene has paved the way for understanding molecular mechanisms involved in nutrient mobilization of essential nutrient (phosphorus) and its subsequent enhanced uptake by plants, as mediated by the rhizobacteria. Multi-location trials were carried out with inoculation of B. licheniformis strain CKA1 to access its effect on early vegetative growth of apple plants under field conditions. Effect of strain CKA1 on vegetative growth of apple was also compared at fresh plantation and re-plantation sites. Results revealed significant increase in vegetative growth over un-inoculated controls at all the locations studied. Effect of liquid formulation of CKA1 on apple yield, pomological traits, and fruit content and nutrient parameters was also evaluated at different locations. Comparison of different application strategies of liquid formulation viz. soil drenching in basin of apple trees alone, foliar spray at pink bud stage and soil drenching in combination with foliar spray for three consecutive years revealed that maximum apple yield/tree (28.00kg at RHRS, Mashobra; 103.75kg at Sablog, Matiana; 20.23kg at Nanni, Matiana; 110.50kg at Kyari, Kotkhai) was observed when apple trees were treated with CKA1 using a combination of root inoculation and foliar spray. Population dynamics studies carried out to determine the fate of inoculated strain of CKA1 in apple rhizosphere revelled successful establishment and colonization of the inoculated strain as indicated by high values Simpsonâs index of Dominance which ranged from 1.51 to 3.40 at different locations. Apple orchard rejuvenation projects are being ambitiously launched by State agencies for restoring six decade old planted apple orchards. Integrated nutrient management systems comprising biological systems especially, single strain multifunctional PGPR such as strain CKA1 opens up new avenues not only for improving crop yield but also in sustaining soil health.ThesisItem Open Access EVALUATION OF POTENTIAL OF LACTIC ACID BACTERIA ISOLATED FROM DAIRY PRODUCTS OF HIMACHAL PRADESH FOR THEIR EXOPOLYSACCHARIDE PRODUCTION AND ITS APPLICATION(2016) SHARMA, KANIKA; SHARMA, NIVEDITAABSTRACT The present investigation was carried out to isolate most efficient potential lactic acid bacteria from dairy products of Himachal Pradesh were screened exopolysaccharide (EPS), identification, safety assessment, evaluation of probiotic attributes, optimization, characterization as well as development of purified EPS based food product. Sixty eight potential lactic acid bacteria were screened for exopolysaccharide (EPS) production by using ruthenium red MRS agar. Out of these 68 isolates, eight i.e. KM0, KM1, KL14, KL15, KM4, JC5, KM3 and KC7 were found positive for EPS production. The quantitative analysis for EPS production was done by using phenol sulphuric acid method and KM0 showed highest EPS production (4.33 mg/ml), followed by KM1 (3.46 mg/ml) and KL14 (2.31 mg/ml). Isolates KM0, KM1 and KL14 were selected for further studies having maximum EPS production, which were isolated from Milk cream, Milk and Lassi, respectively that are dairy products of Himachal Pradesh. Isolates KM0, KM1 and KL14 were identifies as Pediococcus acidilactici, Lactobacillus paraplantarum and Lactobacillus casei by 16S rRNA gene technique and registered in NCBI under accession no. KX671557, KX671558 and KX774469, respectively. Safety assessment of EPS producing isolates was done by evaluating antibiotic susceptibility, haemolytic, DNase and gelatinase activities. All the isolates exhibited 80-100% antibiotic sensitivity, non-heamolytic, non-DNase and non-gelatinase activities, thereby proving their safe status. These screened isolates were further examined for their probiotic potential viz. acidity tolerance, bile tolerance,auto aggregation, co-aggregation, hydrophobicity, and cumulative probiotic potential. All the three screened exopolysaccharide producing isolates were found to be highly acidity tolerant strains with 93.65-99.41 % survival rate at pH 3 for 3 h. These three isolates were able to resist high bile salt concentration i.e. 2.0% with 93.12-98.22 % survival rate for 8 h. All the three isolates exhibited good autoaggregation capacity i.e. greater than 40% after 5 h and showed string hydrophobicity towards xylene i.e. > 40%. The entire screened three isolates were highly qualified with cumulative score of 95.83%.Five parameters i.e. incubation time, temperature, pH, carbon source, surfactants and nitrogen concentration were optimized by using One Factor at a Time approach (OFAT) followed by Response Surface Methodology (RSM) for EPS production. The best carbon source was lactose (1.5%, 2% for KM0 and KM1) while sucrose for KL14 (2%) and best nitrogen concentration was (0.2 % for KM0 and KM1, 0.3% for KL14). The optimum pH, temperature and incubation time were 6.50, 35ÂșC, respectively for all the isolates. Under optimized conditions, an overall increase of 34mg/ml, 32 mg/ml and 28 mg/ml EPS production was observed with KM0, KM1 and KL14, respectively. A step by step purification of exopolysaccharide was done by P. acidilactici KM0, L. paraplantarum KM1& L. casei KL14 , their characterization of KM0, KM1& KL14 was done and the purified exopolysaccharide showed wide stability in high temperature(heat shock & cold shock), pH, storage stability, waterholding capacity, thin layer chromatography (TLC). Among all the isolates, L. paraplantarum KM1 showed maximum EPS production, so it was selected for further study. KM1 isolate characterized by HPLC, MALDI-TOF and SEM. Further, the purified exopolysaccharide was used for the curd formation. The exopolysaccharide was found effectively active to be used in dairy industry as it showed 63.42 % water holding capacity, improve taste and texture of curd.