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ThesisItem Open Access In Vitro Multiplication Of Oriental And Asiatic Hybrids Of Lilium(Chaudhary Charan Singh Haryana Agricultural University; Hisar, 2006) Krishnarajah, Shelomi A.; Chowdhury, V.K.ThesisItem Open Access Identification Of Sex-Linked Dna Markers In Jojoba (Simmondsia Cbinensis (Link) Schneider)(Chaudhary Charan Singh Haryana Agricultural University; Hisar, 2006) Sharma, Richa; Chowdhury, Vijay KumarThesisItem Open Access Dna Fingerprinting And Mapping Of Anthracnose Resistance Gene In Sorghum(Chaudhary Charan Singh Haryana Agricultural University; Hisar, 2005) Singh, Monika; Boora, K. S.ThesisItem Open Access Dna Fingerprinting And Phylogenetic Analysis Of Basmati And Other Rice Types Using Ssr And Transposon Element Based Markers(Chaudhary Charan Singh Haryana Agricultural University; Hisar, 2006) Kaushik, Amit; Jain, R. K.ThesisItem Open Access Genetic and microsatellite marker analysis of azucena (japonica) x HBC19(Basmati) F5-F6 rice population(Chaudhary Charan Singh Haryana Agricultural University;Hisar, 2005) Pankaj Kumar; Chowdhury, V. K.ThesisItem Open Access Genetic improvement of Saccharomyces spp. for ethanol production from starch(CCSHAU, 2005) Bajaj, Anshu; Chaudhary, KamlaA study was undertaken to improve ethanol production from starch by yeast Saccharomyces spp. A total of 10 isolates of yeast were isolated. After secondary screening in submerged culture conditions six isolates were selected for further investigation. Based on Maximum (6.4U) amylase and ethanol (8.8g/l) production, the Saccharomyces spp. B6 was selected and mutagenized using UV rays and nitrous acid. The Saccharomyces spp. mutant B6U3 produced maximum amylase (8.9U) & ethanol 16.4g/l (in starch) and 26.8g/l (in dextrose) among various mutants and was selected for further studies. The optimization of culture conditions showed that Saccharomyces spp. B6U3 produced increased level of amylase as compared to parent strain. Ethanol production was found to increase with increase in the level of starch. At high concentration of starch the ethanol production was decreased. Starch at a concentration of 10% was found be the best for production of ethanol by Saccharomyces spp. B6U3. Peptone was found to be best nitrogen source for production of ethanol by Saccharomyces Spp. B6U3 producing a maximum 33.4 g/l of ethanol in 48h. In media containing ammonium sulphate, ammonium nitrate casein & urea less amount of ethanol was produced. Peptone was found to the best nitrogen source at 2% level and 34.4g/l of ethanol was produced in 48h of fermentation. Increase in the conc. of peptone led to decreased ethanol production. On increasing the inoculum size upto 20% the ethanol production also increased from 5.6g/l to 32.8g/l after 48h of fermentation. A little variation was found on using 20% of starch grown and dextrose grown inoculum. There was a slight increase in the ethanol production on using dextrose grown inoculum (34.8g/l) than starch grown (33.2g/l) after 48h of fermentation. Ethanol production was found to be maximum after 48h of fermentation. Out of different starch sources used Soluble starch, Sorghum, spoiled wheat, Potato and Cassava, soluble starch was found to be the best substrate for production of ethanol by Sacchromyces spp. B6U3.ThesisItem Open Access Cellulase synthesis in 2-deoxy glucose resistant mutants of trichoderma reesei p-2(CCSHAU, 2005) Tina Mittra; Chaudhary, KamlaA study was under taken to isolate 2-DG resistant mutants of Trichoderma reesei P-2 for cellulase synthesis. Spores of T. reesei were mutagenized with UV irradiation for a time period of 15 minutes. At this time maximum number of mutants were obtained.Antimetabolite resistant mutants were obtained on 0.05% 2-DG containing Reese medium plates. A total of 24 isolates which grew profusely, were purified and again patched on Reese medium contained 2-DG. Finally 11 mutants showed good growth on this medium. 2-DG resistant mutants of T. reesei mutants MU-22 and MU-24 showed maximum cellulase production and selected. T.reesei mutants MU-22 and MU-24 showed maximum FPase activity (1.54 IU/ml and 1.56 IU/ml) respectively, CMCase activity (6.4IU/ml and 5.5IU/ml) respectively in cellulose medium. However and -glucosidase activity was more in MU-22(0.28 IU/ml). Effect of different carbon sources on synthesis of cellulases were analysed in mutants MU-24 and MU-22. Cellulose was found to be the best carbon source whereas in cellobiose and cotton medium, cellulase activity was reduced. In phosphocellulose medium -glucosidase activity was high and sorbitol was not a good inducer for the synthesis of cellulases. The optimization studies for cellulase production by MU-24 showed that cellulose at 1% concentration produced high level of cellulases. Similarly ammonium sulphate at a concentration of 0.14% was found to be essential for all the three enzyme activity. Since T. reesei is a mesophilic organism, at high temperature it did not grow. The optimum temperature for growth of T. reesei was found to be 30ºC. Saccharification of rice straw indicated the release of 50% reducing sugar which were more (35%) then the parents strain. The protein profile of culture filtrate of T. reesei mutants were determined by SDS PAGE. It was observed that the first set of proteins which appeared during cellulase induction were of 50KDa, 40KDa, 30KDa and 25KDa. Thus from this investigation it can be concluded that level of exoglucanase, endoglucanase and -glucosidase has been altered by mutation and such improvement in altering the level of individual component of cellulase complex is practically possible.ThesisItem Open Access Fingerprinting satawar (Asparagus racemosus) genotypes using RAPD markers(CCSHAU, 2005) Arora, Puneet; Dhillon, SantoshSatawar (Asparagus racemosus) is a medicinal plant growing in tropical climates and is useful in curing a wide array of ailments. This study was thus undertaken to prepare a DNA fingerprint database of selected varieties of Asparagus racemosus and to assess genetic diversity among them using randomly amplified polymorphic DNA (RAPD) markers. Twenty five RAPD primers were used to assess molecular polymorphism in fifteen Asparagus racemosus genotypes. A total of 211 amplified products were obtained out of which 50 were monomorphic and 161 were polymorphic. Average polymorphism across fifteen genotypes was found out to be 78.650%. For the genotypes tested, 5 to 17 bands were obtained, with an average of 10.55 bands per primer. The size of amplified fragments ranged from 230-2250 bp. Some primers also produced unique alleles in specific Asparagus genotypes which could be used to distinguish them. Analysis of this polymorphism profile, generated using suitable statistical programmes, grouped the fifteen genotypes into two major clusters at a similarity coefficient of 0.680. Varieties HAR-6 and Wild-3 were found out to be the most diverse and distant from other varieties. The second cluster again divided into two minor clusters, out-grouping the Nepali variety. The next large sub-cluster contained all the other genotypes. Varieties Wild-1 and Indian Yellow were genetically most similar. Genetic similarity matrices of the genotypes ranged from 0.630 to 1.00, indicating a moderate genetic variability among the genotypes. Wild-1 and Indian Yellow, showed a genetic similarity value of 1.00 while the genotypes HAR-1 and Wild-3 were found out to be genetically most diverse, at a value of 0.630. All other genotypes varied between these two extreme values. The results indicated that RAPD markers are efficient for identification of Asparagus racemosus genotypes and for determination of the genetic relationships among them. Fingerprint data obtained in this study can be further utilized in identification and development of improved Asparagus varieties.ThesisItem Open Access Identification of gene for ADP-Glucose pyrophosphorylase from developing grains of wheat(CCSHAU, 2006) Saha, Abhigyan; Sikka, Virendera K.A step towards isolation of gene(s) has been undertaken in the proposed study. In order to achieve the target of AGPase cDNA gene library preparation, first of all total RNA was isolated from developing grains (14 DAA), of wheat varieties C306 and WH711. Modified RNA extraction protocol was employed, which gave fairly high quality RNA. Good quality mRNA was fractionated from total RNA using poly AT tract magnetic beads mRNA isolation system (Promega). Complementary DNA (cDNA) library was generated from the mRNA of C306 and WH711 using Lambda zap cDNA kit (Stratagene). Plaque lifts were hybridized with rice AGPase gene probe to identify clones containing wheat AGPase gene. However, exact clone could not be isolated. AGPase enzyme assay was carried out in wheat varieties C306, WH711, WH147M, SG70, MLU-2, at 14DAA and 28DAA. The enzyme activity showed an association with rate of starch accumulation and grain size and yield.
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