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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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Subject: Microbiology × End date: 2018 × Start date: 2018 × Type: Thesis × Thesis Type: Ph.D ×
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    ThesisItemOpen Access
    Characterization of sulphur oxidizing bacteria and their effect on growth of mustard (Brassica juncea L.)
    (CCSHAU, 2018) Chaudhary, Suman; Goyal, Sneh
    Sulphur is a vital element for plants next to nitrogen (N), phosphorus (P), and potassium (K). It is an important constituent of proteins, enzymes, vitamins, lipids, carbohydrates, and other biomolecules. Sulphur is required for growth and development of plants especially in the crop production. Sulphur undergoes a number of biological alterations in nature carried out exclusively by microbes through sulphur cycle. Oxidation of sulphur is the most important step of S cycle which improves soil fertility. It results in the production of sulphate, which can be easily used by the plants, while the acidity produced by oxidation step helps to solubilize plant nutrients and thus improves soil health. Sulphur deficiencies in soils of tropical and subtropical regions have been recognized for many years and have been reported from over 70 countries, including India. Pyrite is good source of sulphur used as sulphur fertilizers and it can be easily used by sulphur oxidizing bacteria. In the present investigation, sulphur oxidising bacteria were isolated from different places of Haryana and evaluated for their effect on growth of mustard. A total of 130 bacteria were obtained, out of which 46 were selected on the basis of dye reduction test and further screened for sulphate production. Five bacteria namely SSD11, SSR1, SSG8, SSF17 and SSH10 were selected on the basis of maximum sulphate production for further studies and were optimized for laboratory cultural conditions. The period of 48 hrs. was found optimum incubation time for all bacteria and 30 oC was best temperature for the growth of SSD11, SSR1, SSG8, SSH10 while 35 oC was for SSF17. The pH 8 was found best for all four isolates except for SSF7, which was 6, while media having glucose carbon source was producing maximum sulphate. The best nitrogen source was found to be ammonium sulphate. After optimization, the selected bacterial isolates were evaluated for their effect on growth of mustard under pot house. There was a significant increase in the height, weight, no. of siliquae, 100 seed weight, oil content, leaves protein content, leaves chlorophyll content, viable rhizospheric bacterial count with the inoculation of bacterial isolates in comparison to control. The selected bacterial isolates were characterized on the basis of morphological and biochemical characteristics. After sequencing of 16S rRNA of bacterial isolates or molecular characterization, the four bacterial isolates were recognized as SSF17 (Phytobacter diazotrophicus), SSR1 (Enterobacter cloacae), SSD11 (Enterobacter cloacae) and SSG8 (Klebsiella oxytoca).
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    ThesisItemOpen Access
    Bioherbicidal potential of antagonistic rhizosphere bacteria in management of wild oat (Avena fatua L.) and their inoculation effect on growth of wheat (Triticum aestivum L.)
    (CCSHAU, 2018) Dahiya, Anupma; Sindhu, S.S.
    In the present study, seventy five rhizobacterial isolates were obtained on the basis of morphological characteristics from wheat rhizosphere soil. These rhizobacterial isolates were screened for antagonistic interaction along with 13 reference strains against Rhizoctonia solani and Neovossia indica under in vitro conditions. Twenty three isolates showed antagonistic activity against R. solani and 33 isolates showed antagonistic activity against N. indica. Isolates BWA36, RWA42, RWA48, RWA53, HCA3, HCA61 and RCA3 showed significant growth inhibition against R. solani, whereas isolates BWA6, BWA19, BWA23, RWA48, RWA53, HCA61 and RCA3 showed the inhibition against N. indica. Thirty four selected antagonistic bacteria were studied for their effect on root and shoot growth of Avena fatua seedlings on 0.8% water agar plates. Eight rhizobacterial isolates BWA6, BWA19, BWA29, RWA48, RWA55, RWA63, RWA71 and HCA61 showed root growth inhibition at both 5th and 10th day of seed germination. Bacterial isolates i.e. BWA6, BWA19, BWA29, BWA38, RWA48, HCA61 and JMM24 caused shoot growth inhibition at both 5th and 10th day of seed germination of A.fatua. Culture filtrate of selected cultures were sprayed on 1-week and 2-weeks old Avena fatua and wheat seedlings. Isolates BWA25, BWA29, RCA3 and SYB101 caused yellowing of leaves, whereas two rhizobacterial isolates BWA18 and RWA52 caused appearance of disease spot on weed. IAA production in selected rhizobacterial isolates and it varied from 3.49 to 53.80 μg/ml. Significant ALA production (> 7 μg/ml) was observed in bacterial isolates BWA25, HCA61, RCA3, HCA3 and SYB101. ACC utilization was observed in 85.29% bacterial isolates and five isolates i.e., BWA20, BWA23, BWA29, BWA38 and RCA3 showed significant growth on ACC supplemented plates. Four bacterial isolates BWA25, BWA52, RWA53 and RCA3 showed significant HCN production. Ten bacterial isolates i.e., BWA2, BWA18, BWA25, BWA29, RWA48, RWA52, RWA69, SYB101, HCA61 and RCA3 were selected on the basis of different beneficial properties to study their inoculation effect on growth of wheat and weed under pot house conditions. Rhizobacterial isolates BWA18, RWA52, RWA69 and SYB101 stimulated growth of wheat and rhizobacterial isolates i.e., BWA18, BWA29 and RWA52 inhibited the growth of A. fatua in comparison to RDF amended uninoculated soil treatment. At 25 days of observation, bacterial isolate BWA18, RWA69, SYB101 showed significant increase in root dry weight (RDW) and shoot dry weight (SDW) of wheat, whereas its inoculation decreased RDW and SDW of A. fatua. At 50 days of observation, inoculation of bacterial isolates BWA18 and RWA48 increased RDW and SDW of wheat and weed both. Isolates RWA69 and SYB101 showed significant increase in RDW and SDW of wheat, whereas its inoculation decreased RDW and SDW of A. fatua. At 75 days of observation, inoculation with bacterial isolates RWA52, RWA69 and SYB101 caused significant increase in RDW and SDW of wheat, whereas its inoculation decreased RDW and SDW of A. fatua as compared to RDF amended uninoculated soil. Rhizobacterial isolates BWA18, RWA52 and RWA69 were identified as Acinetobacter variabilis, Bacillus siamensis and Bacillus endophyticus, respectively by the 16S rRNA sequence analysis. These bacterial isolates could be further exploited as bioherbicide for wild oat and growth improvement of wheat under field conditions.
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    ThesisItemOpen Access
    Impact of municipal solid waste on soil microbiological properties and wheat (Triticum aestivum L.) growth
    (CCSHAU, 2018) Dhanker, Rinku; Goyal, Sneh
    With rapid growth in population and industrialization, municipal solid waste (MSW) generation has been escalating day by day. It is a solid residual product, which is generated from the municipal wastewater treatment plants containing huge quantities of organic matter, micro and macronutrients and some of the trace elements. The direct application of MSW lead to build up the soil organic matter and may stimulate the plant growth. However, MSW may contain some heavy metals and pathogens. While some heavy metals are necessary for the living organisms in trace amounts, most are harmful and hazardous in high quantities. The repeated application of MSW may result in the accumulation of these heavy metals to such an extent, which may prove detrimental to the plants and microbes. In the present study, two different municipal solid wastes were collected from sewage treatment plants situated at CCS HAU Hisar (MSW1) and Hisar city (MSW2) Haryana and analysed for organic matter, different plant nutrients (total N, P, C and K) and heavy metal concentrations. The heavy metal contents were below the EU permissible limits. The DTPA extractable forms of heavy metals were more in solid waste collected from CCS HAU Hisar (MSW1) as compared to the solid waste collected from Hisar city (MSW2). These MSW were added to the soil at the rates of 5, 10, 20, 30 and 50 t/ha and incubated under laboratory conditions upto 90 days. Sub samples were drawn at different time intervals and analysed for different chemical and microbiological properties of soil. There was significant increase in soil organic C, total N and EC over control. Soil organic C gradually declined with incubation time, however no significant difference was observed in total N after 90 days. The DTPA extractable forms of heavy metals (Pb) increased upto 15 and (Ni, Cr, Cu and Cd) 30 days of incubation and then declined with further incubation of 90 days. Microbial biomass C, N, soil dehydrogenase, alkaline phosphatase, urease and cellulase activities, increased significantly in soil amended with different levels of both MSW. Total bacterial and E. coli count increased with increasing levels of solid waste throughout the 90 days of incubation. Fungal and phosphate solubilizing bacterial count increased upto 30 days, however, population of Azotobacter increased with increasing application rates of MSW1 and MSW2, but no significant difference was observed with incubation period. Addition of the MSW1 and MSW2 to the soil promoted the functional diversity (catabolic potential) of the soil microbial population. Under pot house conditions the dry matter yield and uptake of N, P, K by wheat crop increased significantly with increasing application rates of MSW1 and MSW2 from 5 to 50 t/ha, over the control. However, the yield and nutrient uptake by mustard crop increased with application of MSW upto the level of 20 and 30 t/ha of MSW1 and MSW2, which further declined with 50 t/ha of solid waste. The accumulation of heavy metals (Cr, Cd, Ni and Pb) in Indian mustard crop increased significantly with increasing levels of MSW1 and MSW2 and was more than wheat crop.
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    ThesisItemOpen Access
    Development of promiscuous and effective rhizobia nodulating kharif legumes
    (CCSHAU, 2018) Subha; Gera, Rajesh
    Clusterbean, cowpea, mothbean, mungbean, pigeonpea and urdbean are the commonly grown kharif legumes of arid and hyper-arid zones. Inoculation of effective and promiscuous rhizobia improves the nitrogen fixation as well as plant biomass yield of legumes. So, the present investigation has been undertaken for development of promiscuous and effective rhizobia nodulating kharif legumes. A total 209 rhizobial isolates were isolated from 31 soil samples collected from different districts of South-West Haryana and Rajasthan. All rhizobial isolates were checked for their nodulation ability and efficiency in six different kharif legumes. Three rhizobial isolates (PKR177, PKR188 and PKR194) have ability to nodulate all these six crops but with poor nodulation efficiency. So, these three rhizobial isolates were mutagenised by EMS (0.15%) for enhancing nodulation efficiency in all six legumes and it was found that, out of 300, only 4 rhizobial mutants (PKR177m30, PKR188m85, PKR194m24 and PKR194m93) exhibited increase in nodulation efficiency in all the legumes. Rhizobial mutant PKR188m85 was tagged with gfp gene to check its nodulation occupancy in all the legumes. Twenty five native rhizobial isolates, 4 mutants and one GFP marked strain having capability of nodulating more than three crops were evaluated for different traits. Most of the native and mutant rhizobia were able to grow up to 40oC, 4% salt and 30% PEG concentration. All rhizobia were able to produce IAA and ammonia, however, 47 and 63% rhizobia had the ability of bacteriocin production and phosphate solubilization, respectively, whereas, 83% of rhizobia were able to utilize ACC. Most of rhizobia were found to have intrinsic resistance to ampicillin, chloramphenicol, nalidixic acid and streptomycin upto concentration of 100 μg/ml. All the rhizobia showed the presence of nodC gene, however, nifH gene amplification was observed in 26 rhizobia. ARDRA results showed that all the isolates were distributed in to two major groups with sub and sub groups and divergence among these started at 63 percent similarity with restriction enzymes, MspΙ and HaeΙΙΙ. Rhizobial mutants PKR177m30 and PKR188m85 showed better nodulation efficiency and plant growth parameters than control in chillum jar and pots under both sterilized as well as unsterilized conditions. It was observed that promiscuous GFP marked strain showed more than 80% nodule occupancy under unsterilized conditions.The promising promiscuous rhizobial mutants were identified as Rhizobium sp. on the basis of ribo-typing of partial 16S rRNA gene.
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    ThesisItemOpen Access
    Genetic diversity of rhizobia nodulating sesbania cultivars in Indian soils
    (CCSHAU, 2018) Kuldeep; Gera, Rajesh
    The genus Sesbania contains about 70 species widespread over tropical and subtropical regions. The genus has attracted interest for its fast growth, high crop yield, flood tolerance, root as well as stem nodulation with high nitrogen (N2) fixation activity. Due to their importance and diversity within natural populations, identification and characterization of rhizobial strains imply a comprehensive and accurate approach. Yet little is known of the rhizobia that associate with different Sesbania species in terms of their distribution, diversity, ecology or taxonomic status. Therefore, the present investigation was undertaken for genetic diversity of rhizobia nodulating Sesbania cultivars in Indian soils. A total of 70 rhizobial isolates of different Sesbania species were isolated from 56 soil samples collected from different locations of India using trap plant method. These mainly include 14 isolates of S. sesban, 20 of S. grandiflora, 10 of S. aculeata, 10 of Sesbania rostrata (root nodulating) and 16 of S. rostrata (stem nodulating). The authenticity of these rhizobial isolates were tested on the basis of Gram‟s staining, Hofer‟s alkaline medium, Ketolactase and acid or alkaline production tests. All rhizobia were able to produce IAA and ammonia, however, 79, 40 and 27% rhizobia had the ability of phosphate solubilization, siderophore production and bacteriocin production, respectively, whereas, 54% of rhizobia were able to utilize ACC. On the basis of different PGP traits and nodulation efficiency under aseptic controlled conditions, 20 promising rhizobial isolates were selected. Most of these isolates were found to have intrinsic antibiotic resistance to ampicillin, chloramphenicol, nalidixic acid and streptomycin upto concentration of 100 μg ml-1. The Genomic DNA of selected rhizobial isolates for different Sesbania species was amplified for 16S rRNA gene using BAC27F and BAC1378R primers using standard PCR amplification conditions. ARDRA results showed enormous diversity among themselves and divergence among different Sesbania rhizobial isolates was initiated at approximately 54-65% levels of similarity coefficient. These isolates were distributed into two major groups and further into various sub and sub-sub groups. Sole carbon source utilization pattern (SCSUP) with 22 different sugars of 20 selected rhizobial isolates showed diverse sugar utilization pattern. Most of the isolates (88%) utilized dextrose as carbon source, while inulin was the least utilized source of carbon. Five rhizobial isolates; SSKr(ii), SGKe(i), SAMa, SRKr(i)/r and SRTn/s were found to have more nodule number and nodule fresh weight as compared to control and remaining isolates both under sterilized as well as unsterilized conditions. These rhizobial isolates also showed significant increase in shoot and root dry weight along with shoot N and P uptake in their respective crop tested. These isolates have tremendous potential in near future to be used as biofertilizers in salt affected, alkaline and waterlogged field conditions, which will not only improve nitrogen availability and plant growth in Sesbania spp. but also act as nitrogen reserve for next crop. The promiscuous Sesbania rhizobial isolates were identified as Rhizobium sp. on the basis of ribo-typing of partial 16S rRNA gene.
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    ThesisItemOpen Access
    Evaluation of bacterial endophytes for growth promotion of mungbean (Vigna radiata (L.) Wilczek)
    (CCSHAU, 2018) Rekha; Leela Wati
    Mungbean (Vigna radiata (L.) Wilczek), also known as green gram, is one of the important pulse crop of India. India alone accounts for 65% of its world acreage and 54% of the production. Nonrhizobial endophytic bacteria exert several beneficial effects on host plant such as stimulation of plant growth, nitrogen fixation and induction of resistance to plant pathogen. The use of endophytes offers an attractive way to replace chemical fertilizers, pesticides and supplements. In the present study, Mungbean nodule samples were collected from farm area of Pulses section, Genetics and Plant Breeding, CCS HAU, Hisar during summer and kharif season to isolate the endophytes. In total, 41 endophytes were isolated using tryptone soya agar (TSA) medium. After isolation, all the endophytes were assessed for temperature tolerance (up to 450C) and salt (NaCl) tolerance (up to 5%). The majority of mungbean endophytes were found to be IAA producers, though the production varied in the range of 1.12-44.88 μg/ml and the P-solubilisation activity of endophytic isolates on solid as well as liquid medium was restricted to 49 % of the isolates only. Ammonia excretion varied in the range of 0.25-3.47 μg/ml and isolate E17 exhibited maximum ammonia excretion (3.47μg/ml). Out of 41 isolates only 10 endophytic isolates were siderophore producers and 12 endophytic isolates showed HCN production and overall 76% of mungbean nodule endophytic isolates showed ACC utilization on minimal medium plates. Potassium solubilisation was shown by only 24% of the endophytic isolates. Four promising endophytic isolates, namely E6, E13, E14 and E17 which showed maximum number of plant growth promoting traits were selected for plant growth evaluation of mungbean under pot house conditions during summer and kharif season (2016). Among various treatments, coinoculation of endophytic isolate E13 along with Rhizobium (MB703) and phosphate solubiliser (P36) was the best treatment with respect to nodulation, plant growth and yield attributes under pot house as well as field conditions. On the basis of biochemical characterization and partial 16S rDNA sequencing isolates E6, E13, E14 and E17 were identified to be Pseudomonas sp. strain PM1220, Bacillus amyloliquefaciens strain PM1218, Bacillus flexus starin PM1217and Beijerinckia fluminensis strain PM1219.