Loading...
Thesis
Browse
12 results
Search Results
ThesisItem Open Access Genetic Investigations on Heat Adaptive Traits in Bread Wheat by Using Molecular Markers(University of Agricultural Science, Dharwad, 2016-11) Jaya; Biradar, Suma S.ThesisItem Open Access Evaluation and Characterization of Genetically Modified Cotton G. herbaceum Var. Jayadhar for Helicoverpa armigera Resistance(University of Agricultural Science, Dharwad, 2016-11) Mahawar, Sonam; Katageri, I.S.ThesisItem Open Access Marker Aided Introgression of Blast Resistance Genes Pi1, Pi2 and Pi54 Into Intan Rice Variety(University of Agricultural Science, Dharwad, 2016-12) Debnath, Prasenjit; Prashanthi S.K.Rice blast caused by Magnaporthe oryzae is the most devastating fungal disease that causes approximately 80 % yield loss. Use of resistant cultivars is the most effective and economical way to control rice blast disease. “Intan” is a medium slender indica variety, popular with farmers and consumers in Karnataka but highly susceptible to blast disease. BPT5204 NIL-28, 18, 30 introgressed with Pi1, Pi2 and Pi54 which was developed in the Department of Biotechnology, UAS, Dharwad and Tetep, having Pi1 and Pi54 were used as donor parents in the crossing programme to develop F1’s, BC1F1’s and BC2F1’s. Marker assisted backcross breeding approach was adopted to introgress broad spectrum blast resistance genes Pi1, Pi2 and Pi54 independently and pyramided into Intan in 2015 & 2016. Molecular markers genic/linked, flanking and unlinked to target genes were used as foreground, recombinant and background selection markers respectively. Genic marker RM224 for Pi1 gene, tightly linked marker AP5659-5 (0.10 cM) for Pi2 gene, RM206 (0.6 cM) for Pi54 gene showed polymorphism among the parents. These polymorphic markers were employed to confirm target genes in hybrids and backcross population. F1 plants generated from ‘Intan x Tetep’ cross was confirmed for the presence of Pi1 + Pi54 genes and confirmed plants were backcrossed. Further, these F1 plants were challenge inoculated with Magnaporthe oryzae isolates and F1’s showed resistant reaction, confirming the hybridity. Foreground selection was exercised and heterozygous plants with Pi1, Pi54, Pi1 + Pi54 pyramids were identified in BC1F1 generation. Sixty three genome wide markers were subjected for polymorphism and polymorphic markers were employed for background selection. From ‘Intan x BPT5204 NIL-18’ cross, heterozygous plants for Pi2 gene were confirmed in BC2F1 generation and subjected for background selection, in which recurrent parent genome recovery ranged from 57.40 % to 81.48 %.ThesisItem Open Access Pyramiding of Durable Leaf Rust Resistance Genes in Popular Bread Wheat (Triticum aestivum L.) Variety C-306 Through Marker Assisted Backcrossing(University of Agricultural Science, Dharwad, 2016-06) Sowmya G.S.; Desai, S.A.The present investigation was undertaken to pyramid two effective durable leaf rust resistance genes Lr34 and Lr46 for genetic enhancement and improvement of a bread wheat variety C-306. HD2189 carries both Lr34 and Lr46 genes were used as donor for introgression of these two genes employing marker assisted backcrossing. The F1 hybrids of the cross between C-306 x HD-2189 were validated for the presence of both slow rusting genes Lr34 and Lr46, with the help of already reported molecular markers viz., SSR marker cslv34 for Lr34, STS marker Xwmc44 for Lr46 genes, respectively. The obtained BC1F2 and F3 populations were subjected to molecular analysis and thus confirmed plants for Lr34 and Lr46 were grouped into four categories as plants carrying Lr34 alone, Lr46 alone, Lr34 and Lr46 and plants lacking both genes to study the influence of these minor genes on yield and yield attributing traits. Individual plants of BC1F2 and F3 were confirmed for the presence of Lr34 and Lr46 using SSR and STS markers. BC1F2 plants with Lr46, and Lr34+Lr46 genes exhibited high frequency of transgressive segregants, for most of the agronomic traits as compared to Lr34 gene alone. In F3 population, the highest transgressive segregants were recorded for the trait seeds per spike for background of Lr46 alone. Positive promising segregants for both slow rusting genes Lr34 and Lr46 will be the source for durable leaf rust resistance breeding through molecular/marker assisted selection (MAS).ThesisItem Open Access Identification of QTLs for Trichome Traits Implicated in Jassid Resistance of Cotton(University of Agricultural Science, Dharwad, 2016-08) Sankeshwar, Mahantesh; Patil, Rajesh S.An investigation was carried out to identify QTLs for trichome traits implicated in jassid resistance of cotton. A set of 200 recombinant inbred lines derived from the cross Gossypium hirsutum var. DS-28 and Gossypium barbadense var. SBYF-425 was evaluated separately under both protected and unprotected conditions for trichome traits study and jassid incidence respectively, during kharif 2015-16 at the Agricultural Research Station, Dharwad Farm, Dharwad. The mean data of leaf pubescence and yield traits were subjected to analysis of variance and QTL mapping. Analysis of variance revealed significant differences among the RILs for leaf pubescence and productivity traits indicating the presence of substantial variation among the recombinant inbred lines. The jassid population and injury grade were significantly and negatively correlated with leaf pubescence traits viz., abaxial leaf pubescence count, leaf mid-rib pubescence count and leaf vein pubescence count. Notably, the trichome length did not show any relation with either trichome density or jassid numbers on the leaf. Seed cotton yield recorded positive phenotypic correlation with number of bolls per plant and boll weight. The genetic linkage map using 178 recombinant inbred lines constructed earlier was used to analyze and map the QTLs for leaf pubescence, jassid injury resistance and yield traits. QTL mapping software QciMapping was used to analyze the data by single marker analysis, simple interval mapping and composite interval mapping. A total of forty two QTLs were detected on eleven chromosomes, each explaining 7.61 to 18.88 per cent phenotypic variation with peak LOD score of 3.04 to 5.79. Out of forty two QTLs, fifteen affected the leaf pubescence traits, three affected jassid injury resistance and twenty four affected the yield and yield contributing traits. These identified markers associated with leaf pubescence and yield traits can be effectively utilized for maker assisted selection after validation.ThesisItem Open Access Validation of Molecular Markers Linked to Powdery Mildew Resistance QTL in Advanced Breeding Lines and Mutant Lines in Mungbean(University of Agricultural Science, Dharwad, 2016-10) Surya Prakash; Bhat, SumangalaThe present study was conducted to evaluate advanced breeding lines and mutant lines of mungbean [Vigna radiata (L). Wilczek] for powdery mildew resistance and to validate with previously reported molecular markers associated with powdery mildew resistance. The field experiment was conducted at the Main Agricultural Research Station, University of Agricultural Sciences, Dharwad during kharif 2014. The molecular experiments were conducted in the Department of Biotechnology. Phenotypic screening of advanced breeding lines and mutant lines along with the respective parental lines confirmed that parental lines BGS 9, Chinamung, DGGV 2, Selection 4, CO 7 and SML668 are susceptible to powdery mildew reaction and TARM 1 and TM-96-2 resistant to powdery mildew. Among the advanced breeding lines only BGT96-7 and BGT96-12 showed resistant reaction and were on par with TM-96-2. However, for other yield related traits such as number of cluster per plant, number pods per plant, number of seeds per pods and 100 seed weight these two lines were not superior to BGS9, the female parent involved in developing these lines. Advanced breeding lines were also genotyped using markers (CEDG259, CEDG304, MBSSR238, CEDG290, DMBSSR199 and CEDG191) already linked to powdery mildew resistant QTLs. None of the markers showed significant association with powdery mildew resistance in the advanced breeding lines used in this study. As they might carry additional QTLs imparting resistance to powdery mildew, BGT96-7 and BGT96-12 can be used as a resistance source while breeding for powdery mildew resistance.ThesisItem Open Access Isolation and Characterization of Bioactive Compounds from Marine Microbes(University of Agricultural Science, Dharwad, 2016-09) Vishwas, M.; Moger, NarayanThe present investigation was carried out to exploit the anti-phytopathogenic property of marine microbes. Several microbial cultures (four hundred thirty) isolated from the marine water and sediment samples from coastal parts are maintained at Department of Biotechnology, UAS, Dharwad. Among these resources sixty one bacterial, fifty fungal and forty one actinomycetes viable isolates were selected for investigation. These isolates were tested for anti-phytopathogenic activity against Sclerotium rolfsii, Fusarium oxysporum, Colletotrichum sp. and Ralstonia solanacearum in vitro. Three isolates (AUDI-172 from bacteria, AUDI-585 from fungi and AUDI-841 from actinomycete) showed good anti-phytopathogenic activity against all phytopathogens. The molecular characterization of potent microbial isolates were done based on 16s rDNA region for bacteria and actinomycete whereas internal transcribed spacer (ITS) region for fungi. BLAST analysis showed bacterial isolate as Enterobacter cloacae, actinomycete as Streptomyces macrospores and potent fungus as Debaryomyces hansenii. To find out bioactive compounds, secondary metabolites from these potent isolates were extracted using ethyl acetate solvent-solvent extraction method and again tested for antimicrobial activity against the phytopathogens. Functional groups of bioactive compounds were characterized by using gas chromatography-mass spectrometry (GCMS) from the crude extract of potent isolates. Twelve different bioactive compounds were identified from the potent isolates of bacteria and fungi. Thirteen bioactive compounds were identified from potent actinomycete. Mostly, secondary metabolites are the products of polyketide synthesis (PKSs) and non-ribosomal peptide synthesis (NRPSs) gene clusters. Degenerative primer pairs were used for the amplification of gene fragments from PKS-1 and NRPS gene clusters. Presence of PKS-1 and NRPS gene fragments were confirmed in the potent isolates. The present study depicts a promising scenario to focus on marine microbial derived bioactive compounds against phytopathogens. In vivo analysis of these bioactive compounds can help to identify novel biocontrol agents against phytopathogens.ThesisItem Open Access Evaluation of Advanced Breeding Lines for Mungbean Yellow Mosaic Virus (MYMV) Resistance in Mungbean [Vigna radiata (L.) Wilzeck](University of Agricultural Science, Dharwad, 2016-08) Ravikumar; Suma MogaliIn the present investigation, eighty nine advanced breeding lines were evaluated for Mungbean Yellow Mosaic Virus (MYMV) resistance during two successive seasons summer2014 and summer 2015, amongst which, twenty fouradvanced lines showed moderate resistance to MYMV. Further, twenty four lines were evaluated in two replications for MYMV resistance during summer season 2015 under protected and unprotected condition. Out of twenty four advanced breeding lines, seventeen were found to be moderate resistant, three moderate susceptible, two susceptible and only one wild relatives of mungbean Vigna trilobata were found to be immune to MYMV during summerseason 2015.The genetic variability was also studied. Among the twenty four advanced breeding lines yield per plant were significantly and positively associated with number of pods per plant and number of seeds per pod. These advanced breeding lines which are high-yielding and also possess moderate resistance to MYMV can be released as new variety after large-scale field testing. The study was also conducted to validate molecular markers that were previously reported to be linked to major locus/QTLs governing MYMV resistance. Thirty markers were used for marker-trait analysis.Among the thirty,ten markers (viz.,CEDG008, CEDG257, CEDG006, CEDG244, CEDG115, CEDG041, CEDG084, CEDG201, CEDG304 and CEDG191) showed parental polymorphism. But these markers did not show association with MYMV resistance in advanced breeding lines. The crosses between MYMV susceptible and MYMV resistant parents were carried to develop mapping population for MYMV resistance. All the F1 plants from the cross between DGGV2 × Vigna trilobata were validated with thirty markers. Only two markers (CEDG006 and CEDG088) showed parental polymorphism in F1s. Two markers were used for molecular confirmation of F1 derived from a cross between DGGV2 × Vigna trilobata.ThesisItem Open Access Selection of Single Chain Fragment Variable Monoclonal Antibody Against Papaya Ringspot Virus Coat Protein(University of Agricultural Science, Dharwad, 2016-06) Chougala, Annasab S.; Moger, NarayanPapaya ringspot virus (PRSV) is an important virus causing the most devastating disease in papaya. The symptoms caused by PRSV are not visible at early stages. This necessitates early diagnosis of PRSV infections before planting papaya seedlings to main field. The most commonly used diagnostic tool for PRSV detection is immunological assays, which is dependent on the availability of highly specific antibody to differentiate the viruses. Production of antibody using phage display technology requires pure protein therefore, PRSV-CP was expressed in bacteria. An 864 bp PCR product containing coat protein coding region of PRSV was amplified using gene specific primers and amplified product was cloned into the pTZ57R/T and further expressed in the pQE30. After transformation the clones were confirmed through PCR, restriction analysis and sequencing. Amplification with expected size of 864 bp and 94 % homology with other isolates showed integrity of the clone. Further, the coat protein appeared to be expressed at 3 hr after induction with 1 mM IPTG. A band of 32 KDa on the SDS-PAGE confirmed that coat protein was fused to the His-tag. The yield of coat protein about 1.2 mg/100 ml was purified using His-tag purification kit. Four rounds of biopanning were performed by coating purified PRSV-CP into the immunotube using Tomlinson library. The fourth biopan reading (1.46) shown higher binding specificity to PRSV-CP. These were subsequently used for production of scFv monoclones against PRSV-CP. Finally, the randomly selected scFv clones were screened by ELISA. The ELISA readings were observed that three clones had higher binding affinity to PRSV-CP. The sequencing of the clones showed 94% homology with the scFv antibody gene (KF732845.1). The selected clone is highly specific to PRSV-CP as there was no cross reaction with banana BBTV, cucumber CMV and groundnut GBNV. Further, the sensitivity test results imply that the developed scFv antibody can detect at the concentration of 10 µg/ml antigen.
