Thumbnail Image

Thesis

Browse

20102
Reset filters
Subject: Agricultural Biotechnology × Author: Mr. B. J. Malviya ×
Reset filters

Search Results

Now showing 1 - 2 of 2
  • Thumbnail Image
    ThesisItemOpen Access
    PHENOTYPIC AND GENOTYPIC DIVERSITY OF DIFFERENT GROUNDNUT (Arachis hypogaea L) ROOT NODULE BACTERIA (Rhizobium)
    (jau,junagadh, 2010-01) Mr. B. J. Malviya; Dr. V. P. Chovatia
    An investigation was carried out to study the diversity of groundnut Root Nodule Bacteria (RNB), isolated from different areas of Junagadh and Rajkot districts, using various phenotypic and molecular techniques. Twenty four RNB isolates of groundnut, comprising 21 RNB isolates from different areas of Junagadh and Rajkot districts, were studied with three standard strains of Rhizobium, collected from National Research Centre for Groundnut (NRCG), Junagadh. The 21 RNB isolates were selected and confirmed as Rhizobium, using several biochemical tests and the pure cultures were preserved on YEMA slants for further study. All the strains were analyzed for phenotypic tests, namely, pH tolerance, salt tolerance, antibiotics resistance profile and carbohydrate utilization pattern. All the RNB isolates and standard strains grew well at the pH 6.5 to 8.5. Most of RNB isolates were able to tolerate higher NaCl but were unable to tolerate higher CaCl2. Out of eleven antibiotics tested, carbenicillin, erythromycin, gentamicin and streptomycin could differentiate RNB isolates into different groups; while polymyxin-B, ampicillin, tetracycline and chloramphenicol were found ineffective in grouping the RNB isolates. All RNB isolates and standard strains were able to utilize most of carbon sources used here. In plant infection (nodulation) test, carried out in vitro (pot study) with groundnut cultivars (GG-20 and TAG-24) and the RNB isolates, viz JND-3, JND-5, JND-9, JND-13, RJ-4, RJ-7 and IGR-6 revealed positive results for GG-20. While JND-1, JND-3, JND-9, JND-10, RJ-2 and TAL-100 revealed positive results for TAG-24. The isolates JND-9, RJ-5, JND-12 were found superior for root length, shoot length and fresh weight in GG-20. While the isolates JND-3, JND-9, JND-13 were found superior for root length, shoot length and fresh weight in TAG-24. The 16S rDNA PCR-RFLP assay clustered all the isolates into four groups. Majority of RNB isolates were distributed district wise in dendogram. Two strain JND-10 and JND-11 were grouped separately, confirming the phylogenetic relationship. IGR-6 was grouped with RNB isolates of Junagadh district. While TAL-1000 was grouped with RNB isolates of Rajkot districts. The nif H gene amplification revealed only single desired fragment in all RNB isolates and standard strains (approximately 781 bp). BOX-PCR clustered all RNB isolates and standard strains into five groups. All the RNB isolates of group I were similar to standard strains TAL-1000 and IGR-6, while all the isolates of group III were similar to standard strain NC-92.
  • Thumbnail Image
    ThesisItemOpen Access
    PHENOTYPIC AND GENOTYPIC DIVERSITY OF DIFFERENT GROUNDNUT (Arachis hypogaea L) ROOT NODULE BACTERIA (Rhizobium)
    (jau,junagadh, 2010-01) Mr. B. J. Malviya; Dr. V. P. Chovatia
    An investigation was carried out to study the diversity of groundnut Root Nodule Bacteria (RNB), isolated from different areas of Junagadh and Rajkot districts, using various phenotypic and molecular techniques. Twenty four RNB isolates of groundnut, comprising 21 RNB isolates from different areas of Junagadh and Rajkot districts, were studied with three standard strains of Rhizobium, collected from National Research Centre for Groundnut (NRCG), Junagadh. The 21 RNB isolates were selected and confirmed as Rhizobium, using several biochemical tests and the pure cultures were preserved on YEMA slants for further study. All the strains were analyzed for phenotypic tests, namely, pH tolerance, salt tolerance, antibiotics resistance profile and carbohydrate utilization pattern. All the RNB isolates and standard strains grew well at the pH 6.5 to 8.5. Most of RNB isolates were able to tolerate higher NaCl but were unable to tolerate higher CaCl2. Out of eleven antibiotics tested, carbenicillin, erythromycin, gentamicin and streptomycin could differentiate RNB isolates into different groups; while polymyxin-B, ampicillin, tetracycline and chloramphenicol were found ineffective in grouping the RNB isolates. All RNB isolates and standard strains were able to utilize most of carbon sources used here. In plant infection (nodulation) test, carried out in vitro (pot study) with groundnut cultivars (GG-20 and TAG-24) and the RNB isolates, viz JND-3, JND-5, JND-9, JND-13, RJ-4, RJ-7 and IGR-6 revealed positive results for GG-20. While JND-1, JND-3, JND-9, JND-10, RJ-2 and TAL-100 revealed positive results for TAG-24. The isolates JND-9, RJ-5, JND-12 were found superior for root length, shoot length and fresh weight in GG-20. While the isolates JND-3, JND-9, JND-13 were found superior for root length, shoot length and fresh weight in TAG-24. The 16S rDNA PCR-RFLP assay clustered all the isolates into four groups. Majority of RNB isolates were distributed district wise in dendogram. Two strain JND-10 and JND-11 were grouped separately, confirming the phylogenetic relationship. IGR-6 was grouped with RNB isolates of Junagadh district. While TAL-1000 was grouped with RNB isolates of Rajkot districts. The nif H gene amplification revealed only single desired fragment in all RNB isolates and standard strains (approximately 781 bp). BOX-PCR clustered all RNB isolates and standard strains into five groups. All the RNB isolates of group I were similar to standard strains TAL-1000 and IGR-6, while all the isolates of group III were similar to standard strain NC-92.