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University of Agricultural Sciences, Bengaluru
University of Agricultural Sciences Bangalore, a premier institution of agricultural education and research in the country, began as a small agricultural research farm in 1899 on 30 acres of land donated by Her Excellency Maharani Kempa Nanjammanni Vani Vilasa Sannidhiyavaru, the Regent of Mysore and appointed Dr. Lehmann, German Scientist to initiate research on soil crop response with a Laboratory in the Directorate of Agriculture. Later under the initiative of the Dewan of Mysore Sir M. Vishweshwaraiah, the Mysore Agriculture Residential School was established in 1913 at Hebbal which offered Licentiate in Agriculture and later offered a diploma programme in agriculture during 1920. The School was upgraded to Agriculture Collegein 1946 which offered four year degree programs in Agriculture. The Government of Mysore headed by Sri. S. Nijalingappa, the then Chief Minister, established the University of Agricultural Sciences on the pattern of Land Grant College system of USA and the University of Agricultural Sciences Act No. 22 was passed in Legislative Assembly in 1963. Dr. Zakir Hussain, the Vice President of India inaugurated the University on 21st August 1964.
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ThesisItem Open Access MOLECULAR CHARACTERIZATION OF YEAST STRAINS FOR EFFICIENT ETHANOL PRODUCTION USING SSR MARKERS(University of Agricultural Sciences, Bangalore, 2009-07-15) AYEESHA, MUNAWERY; HARINIKUMAR, K.M.The study was carried out to isolate yeast strains from their natural habitat and to screen them for ethanol tolerance. A total of 45 yeast strains were isolated from sugar rich sources. Twelve were identified as Saccharomyces spp. Based on colony type and cell morphological characters such as cell shape and budding characters. Saccharomyces spp. were screened for the ability to tolerate different ethanol concentrations from 5-15% growth in different ethanol concentrations varied from one isolate to another. Yeast isolates showed tolerance level from 5-15%. The best strain had 15% ethanol tolerance strain YDE, which showed high tolerance to ethanol and YMS with low tolerance were mutated by UV radiations with time intervals of 1, 3 and 5 mins and through chemical method using Acridine orange and Ethidium bromide at concentrations of 0.25, 0.50. 0.75 and 1.00 g/ml and were subjected to screening under same ethanol concentrations from 5-15%. Mutated isolates showed decreased growth and tolerance under high ethanol stress compared to their original isolates. SSR profiling was employed to characterize yeast isolates. Eleven Saccharomyces spp. were selected and were subjected for SSR analysis using 10 yeast specific SSR primers selected from public domain and constructed by chromos Biotech Company. Showed polymorphism among the isolates. Dendrogram was constructed according to unweighted pair group arithmetic means using STATISTICA software. Cluster analysis revealed major two groups, two isolates formed one group and other nine isolates formed other group. There was no correlation between SSR profiling and physiological screening of yeast isolates for ethanol tolerance.ThesisItem Open Access TRANSFORMATION OF POMEGRANATE (Punica granatum L.) BY AMP (ANTIMICROBIAL PEPTIDE) GENE TO CONFER RESISTANCE AGAINST BACTERIAL BLIGHT OF POMEGRANATE(University of Agricultural Sciences, Bangalore, 2009-07-15) NUNGSHILEPDEN; SUKHADA, MOHANDASPomegranate is a popular fruit crop and is of considerable economic importance. The bacterial blight of pomegranate is becoming a serious problem in major pomegranate growing area in India. All conventional ways and means of controlling this disease have failed. In pomegranate, existing protocols have shown slow response by regeneration and less work has been reported on transformation. Hence, in the present study, a systematic investigation was carried to standardize an efficient in vitro regeneration protocol from different explants of pomegranate cv. Bhagwa, find out the best treatment for faster regeneration and transforming it with AMP gene. As a result, we could able to achieve a reliable and efficient regeneration protocol and standardize an efficient transformation protocol. Agrobacterium tumefaciens carrying gene pCAMBIA construct with the constitutive CaMV35S promoter, AMP gene terminator and nptII selectable marker (Kanamycin resistance), was used for transformation of explants. Putative transformants were identified on selection medium containing kanamycin at different concentration. Transgene insertion and expression at various levels were confirmed using PCR. Out of four putative transformants analyzed, three transgenic plants showed PCR +ve with AMP gene specific primer and further screening is going on using different molecular techniques.ThesisItem Open Access MOLECULAR FINGERPRINTING OF OKRA ACCESSIONS BY DNA MARKERS(University of Agricultural Sciences, Bangalore, 2009-07-15) PRASHANTH KUMAR, G. M.; ANANTHANARAYANAN, T. V.The molecular analysis of genetic diversity and relatedness assumes considerable significance in evolving appropriate strategies for crop breeding. These investigations were focused on the assessment of the genetic diversity in okra genotypes using DNA markers. In the present study, eighty four accessions were studied to know the genetic similarity using RAPD markers. Out of 60 primers screened, 8 gave good amplification and polymorphism. Out of 8 primers, OPF 10 gave the highest discriminative power (%) i.e. 10.20. OPG 10 gave least polymorphism (50%) among 8 primers. Among the eighty-four accessions, eight random primers generated 17 scorable RAPD loci of which 16 RAPD loci were polymorphic (94.11%). The average number of polymorphic bands per primer was 2. endrogram was constructed based on the pooled data using the STATISTICA software. The genetic dissimilarity value in the distance matrix ranged from 0 to 15% suggesting a narrow genetic base within the okra genotypes. Cluster analysis based on 17 number of RAPD polymorphic bands revealed that 84 genotypes were clustered at a linkage distance of 156 units in the dendrogram with IC 282288 and IC-33315 spanning the extremes. The data generated can be used to improve the agronomically important traits of okra. Knowledge on the genetic variability in okra accessions increases the efficiency of germplasm conservation and enables precision breeding for crop improvement. Key words: RAPD, Abelmoschus esculentus L, genetic diversity, DNA polymorphismThesisItem Open Access ISOLATION AND MOLECULAR CHARACTERIZATION OF Bacillus megaterium AND THE RESPONSE OF AEROBIC RICE TO THE ISOLATES(University of Agricultural Sciences, Bangalore, 2009-07-15) REMYA, G. PILLAI; SURESH, C. K.The investigation was carried out to study the molecular characterization of Bacillus megaterium isolated from soils of the ten different agroclimatic zones of Karnataka. A pot culture experiment was carried out to find out the influence of Bacillus megaterium on aerobic rice under greenhouse conditions. Aerobic rice plants inoculated with Bacillus megaterium isolates manifested increase in plant height, number of leaves, number of tillers, biomass and grain weight compared to uninoculated plants. Among the ten isolates inoculated, the Bacillus megaterium isolates form zone 7 (Southern transition zone) recorded significantly high values in almost all growth parameters chosen for the study. Simultaneously biochemical parameters of the aerobic rice inoculated with 10 isolates of Bacillus megaterium was studied. In the inoculated plants the biochemical and chemical parameters like chlorophyll content, nitrogen, phosphorous, and total sugars was higher as compared to uninoculated plants. The diversity of these isolates was characterized by randomly amplified polymorphic DNA (RAPD) marker. RAPD analysis revealed a total of 47 bands, out of which 40 bands were found to be polymorphic. The RAPD marker analysis clearly depicted that all the ten Bacillus megaterium isolates formed two major clusters. Among the two major groups, isolates from zone 3 and zone 4 formed the first group while those from zone 1, zone 2, zone 5, zone 6, zone 7, zone 8, zone 9 and zone 10 formed the second group. The RAPD banding pattern of these isolates could easily distinguish the isolates of different zones. Variation in these isolates was also reflected on the growth and development of aerobic rice plants as discussed above.ThesisItem Open Access IDENTIFICATION OF DNA MARKERS LINKED TO BACTERIAL BLIGHT DISEASE IN POMEGRANATE (Punica granatum L.)(University of Agricultural Sciences, Bangalore, 2009-07-15) AVINASH, K. N.; RAVISHANKAR, K. V.Pomegranate (Punica granatum L.) is one of the oldest known edible fruit of tropical and subtropical regions belongs to the family Punicaceae. Bacterial blight caused by Xanthomonas axonopodis pv. punicae is one of the severe disease limiting crop yield and productivity thus, affecting the cultivation. In Indian Institute of Horticultural Research, Bangalore, attempts are being made to incorporate resistance to popular cultivars through plant breeding methods. ‘Ganesh’ is a popular variety (susceptible to bacterial blight disease) and ‘Daru’ which is a resistant genotypes are being used as parents in breeding programme. A total of 80 F2 populations derived from a cross between ‘Ganesh’ and ‘Daru’ were used to identify molecular markers linked with the resistant trait. Initially we screened ISSR (Inter Simple Sequence Repeats), RAPD (Randomly Amplified Length Polymorphism), SRAP (Sequence Related Length Polymorphism), RGA (Resistance Gene Analogues) and a few designed primers for determining polymorphism among the contrasting genotypes. A total of 41 molecular markers, including 40 RAPD and one ISSR markers were selected. These primers were then used to amplify segregating F2 s from cross `Ganesh x Daru` to develop a preliminary linkage map in Punica granatum L. using Haldane mapping function. In linkage analysis, 35 markers were mapped on 8 linkage groups. The linkage map length varied from 3.3 cM to 39.8 cM. The map covered a total length of 146.3 cM with an average marker density of 24.38 cM between the two adjacent markers. The maximum number of markers, 8 were found on the linkage groups LG2 and LG6. This linkage map developed forms a basis for development of high density mapping in pomegranate. An attempt was also made to identify marker linked to resistance by following Bulk Segregant Analysis. The primer OPD11 amplified band of approximately 1kbp size in resistant bulk and resistant parent `Daru`.ThesisItem Open Access INDUCTION OF SOMATIC EMBRYOGENIC ACTIVITY IN POMEGRANATE (Punica granatum L.) CULTURES OF MATURE ORIGIN(University of Agricultural Sciences, Bangalore, 2009-07-15) HARSHA, A. S.; LEELA, SHAHIJRAMA study on inducing som atic embryogenic activity in pom egranate was carried ou t by using explants of mature origin viz., petal, le a f and anthers/stamens and m olecular profiling o f the embryogenic callus issued from these explants w as carried out by using RAPD m arkers with an objective o f developing standardized protocol for regeneration of pomegranate, to develop cultivars resistant to bacterial blight pathogen (Xanthomonas axonopodis pv. Punicae). U sing conventional breeding procedures, it is difficult to develop resistant cultivars due to time and space constraint. Hence, somatic embryogenesis offers viable protocols since it produces both root and shoot meristems necessaiy for complete plant growth. For inducing somatic embryos, explants of field origin were collected. Initiation of growth and maintenance o f explants w as done on MS and W PM semi-solid media. Callus initiation was seen after 3 and 8 weeks o f inoculation from petal and leaf explants respectively and somatic em bryos were obtained. Response o f anthers/stamens w as seen positively w hich produced embryogenic callus. Shoot apex form ation was seen from petal and leaf derived embryogenic callus 22 w eeks of inoculation. Callus issued from petal and anther was subjected for RAPD marker analysis. 12 different primers OPG -03, O PG -05, OPG -08, OPG - 13, OPG -16 OPG -18, OPI-01, OPI -03, OPI -10, OPI -12, OPI -14 & OPI -18 screened revealed clear amplification between petal derived and anther derived callus from which it was strongly believed that the anther derived callus is o f sporophytic origin.ThesisItem Open Access EVALUATION OF Bt TRANSGENIC BRINJAL (Solanum melongena L.) AND Bt TRANSGENIC TOMATO (Solanum lycopersicum) FOR RESISTANCE TO THE FRUIT BORERS.(University of Agricultural Sciences, Bangalore, 2009-07-15) PURUSHOTHAMA, A.; VAGEESHBABU, H. S.Brinjal and tomatoes are important, popular, solanceous vegetable crops in the Indian subcontinent. Brinjal fruit and shoot borer (Leucinodes orbonalis Guenee) and tomato fruit borer (Helicoverpa armigera Hubner) are the major pests for brinjal and tomato crop plants respectively. There is no source of resistance in these crop plants. Bt transgenic technology offers a sustainable and successful method of managing these pests. Synthetic Bt genes, cry1Aa3 and cry2Aa were used to transform into brinjal (cv. Arka Keshav) and tomato (cv. Arka Vikas). Evaluation of Bt transgenic brinjal and tomato plants was carried out using molecular and insect feeding analysis. NptII specific primer was used in the PCR based characterization of the transgenic plants using total genomic DNA. Qualitative dot-ELISA was used to study expression of Cry1Aa3 and Cry2Aa Bt proteins in the transgenic plants using Bt Xpresstrips® (Desigen, Mahyco, Jalna). In vitro and transgenic nethouse rearing of Leucinodes orbonalis and Helicoverpa armigera was done to maintain and multiply the insect inoculum for the purpose of challenging on to the transgenic plants for phenotyping. A new detached leaf bioassay method was used to challenge the transgenic and control plants using neonate larval stages of Leucinodes orbonalis and Helicoverpa armigera. Transgenic plants expressing Bt proteins and showing different degrees of resistance to the pests were identified, evaluated and used for selfing to get segregating linesThesisItem Open Access GENERATION OF TRANSGENIC GROUNDNUT (Arachis hypogea L.) PLANT USING EPSPS GENE AS A MARKER(University of Agricultural Sciences, Bangalore, 2009-07-10) SHIVAKUMAR, GURABATTI; SHYAMALAMMA, S.Foot-and-mouth disease (FMD) is the most feared, viral disease of cloven-footed animals causing heavy losses to the livestock industry. The conventional vaccines for FMD have their several limitations which includes safety, temperature sensitivity and duration of immunity. Attempts have been made to overcome these limitations using recombinant DNA technology. Amongst the newer vaccines, edible vaccines are cost-effective, easy-to-administer and socio-culturally acceptable. Further, the use of marker gene to identifying transgenics helps in value addition to the product. The detection methods for the available biochemical markers like GUS protein or antibiotic selection are cumbersome. Hence in the present study, the P1-2A gene of FMDV ‘O’ serotype along with the selectable EPSPS was used. The P1-2A gene is coding for structural proteins of FMDV. EPSPS is a key enzyme in the shikimate biosynthetic pathway necessary for the production of aromatic amino acids. The herbicide glyphosate is a competitive inhibitor of EPSPS. Thus, the gene product titrates the herbicide. The EPSPS gene was cloned into pCambia vector by replacing GUS gene. The cloned gene was confirmed by colony PCR, restriction digestion and sequencing. The CaMV P I-2A gene was cloned into pCambia EPSPS vector and confirmed by colony lysis and restriction digestion. The construct was transferred to Agrobacterium and used for transformation of groundnut seeds. A fully matured leaves of groundnut plants were treated with glyphosate at a concentrations of 1, 2, 3 gm per litre of water. The plants that showed tolerance to glyphosate at 3 gm per litre of water were further screened for presence of FMD VP1-2A gene by PCR. Of the 45 transformants, 8 were found positive both for marker gene and FMDV immunogen. The positive plants are to be further evaluated as edible vaccine.ThesisItem Open Access MORPHOLOGICAL AND DNA MARKER ASSISTED DIVERSITY ANALYSIS OF OKRA GENOTYPES (Abelmoschus esculentus (L.) Moench.)(University of Agricultural Sciences, Bangalore, 2009-07-15) SUNIL KUMAR, K. S.; VIJAYAKUMAR SWAMY, H. V.The genotypes exhibited adequate variability for all eleven characters. The analysis of variance indicated the difference among genotypes were found to be significant except fallowing parameters days to 50% flowering, fruit girth, fruit length and mean internodal length. The PCV was higher than GCV for all characters. Higher GCV and PCV values were obtained for Primary branches per plant, Nodes per plant, Plant height and Fruit yield per plant indicating that variation of these characters contributed markedly to the total variability. High heritability coupled with high per cent of mean genetic advance was obtained for characters viz. Days to 50% flowering, Ridges per plant, Fruit girth, Fruit length, Fruit weight and Mean internodal length. Using D2 analysis of Mahalanodis (1936), thirty genotypes were grouped into eight clusters. Among the clusters, cluster I was the largest with fourteen genotypes followed by cluster VII and VIII with three genotypes respectively. Fruit yield per plant had strong and positive association with days to 50% flowering, Plant height and mean internodal length, both at phenotypic and genotypic level. The extent of genetic variability at molecular level in combination with eleven characters was assessed among okra genotypes along with magnitude association with fruit yield and its component characters. The dendrogram was generated based on STATISTICA for AFLP data resulted in 2 major clusters under which 4 minor clusters are obtained. Minor cluster 4 was the largest with 22 genotypes followed by minor cluster 1 and 3 with 3 genotypes each and minor cluster 2 has just one genotype. The minor cluster 1 and 2 contains IC series genotypes. And in minor cluster 3 and 4 with some local genotypes suggesting the similarity between local, KA, PB and DOV series genotypes. Highest 100% similarity was observed between IC43746 and IC43750. Most of the local varieties are grouped under minor cluster 4.