Thumbnail Image

Master Degree Theses

Browse

2010 - 20124
 Reset filters

Settings

Subject: Plant Biotechnology ×

Search Results

Now showing 1 - 4 of 4
  • Thumbnail Image
    ThesisItemOpen Access
    Isolation and Molecular Characterization of Arsenite Tolerant Bacteria
    (Biotechnology N. M. College of Agriculture Navsari, 2010-11) Jain, Raina; Jha, Sanjay
    Two arsenite tolerant bacteria, one from marine and other from non-marine environment, were isolated by enrichment culture method at 28°C in aerobic conditions. Marine isolate, designated as strain ALANG-4 can tolerate 27mM ASIII (arsenite) and non-marine industrial isolate, GIDC-S can tolerate S4mM Asm, in rich medium. Based on 165 rDNA analysis, strain ALANG-4 was identified as Halomonas sp. and strain GIDC-5 as Alishewanella sp. Both strains were also able to oxidize ASIII to As v by the action of arsenite oxidase. ASIII oxidase activity (localized in membrane fraction) was 88.43"M/min/mg protein and 75.D4J1M1minlmg protein, in presence of ImM ASIII, in ALANG-4 and GIDC-5 respectively. Arsenite oxidase gene, aoxB and arsenite transporter genes, arsB and ACR3, were amplified in DNA samples of both strains, using specific degenerate primers for each gene. Specific activities of antioxidant enzymes like CAT, APX, SOD and GST were increased with increasing amount of ASIII in medium. GR activity was inversely proportional to concentration of arsenite in the medium. When I DmM ASIII was present in medium, 75% and 94% inhibition in activity of PDH, and 85% and 94% inhibition in CI-KGDH activity was observed in ALANG-4 and GIDC-5, respectively. Halomonas sp. strain ALANG-4 overcomes the toxic effect of ASIII by action of efficient eflux system, PYC and GST. Also, its TCA cycle operates at low pace . • In case of Alishewanella sp. strain GIDC-5, GDH and MDH activities were increased proportionate to ASIII concentration; suggesting ED pathway and Glyoxylate cycle were operating in this strain in arsenite stress condition. The results of present study suggest the mode of action of arsenite and on-going metabolic processes in Halomonas sp. strain ALANG-4 and Alishewanella sp. strain GIDC-5 to combat the harmful effects of ASIii.
  • Thumbnail Image
    ThesisItemOpen Access
    In vitro Regeneration and Genetic Transformation of Castor (Ricinus Communis L.) Genotype Skp-84
    (Biotechnology Department, N. M. college of Agriculture Navsari Agricultural University, 2011-11) Kansara, Rohan Vijaykuamr; Jha, Sanjay
    An efficient in vitro plant regeneration and Agrobacterium tumefaciens mediated genetic transformation protocol has been described for castor (Ricinus communis L.) using meristem as explants. Shoot apex containing apical meristems were excised from 5-7 days old in vitro grown seedlings and cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of cytokinins singly or in combination. Kinetin had marked effect on shoot initiation and shoot quality. Kinetin (0.50 mg/l) in combination with BAP (0.25 mg/l) produced maximum number of shoot (10.33) and shoot length (5.20 cm). For root induction. in vitro produced shoots were transferred to rooting media containing half strength MS basal media with NAA. In Agrobacterium tumefaciens mediated genetic transformation, Agrobac/erium tumefaciens strain containing construct pBIN 1 F harboring nptIl and cry] F gene was used. The integration of the gene was confirmed by using peR. The transformation experiment was performed by optimizing two important parameters: age of the seedlings and co-cultivation duration with Agrobac/erium. When the effect of the age of the seedlings and co-cultivation duration were evaluated fifteen days old co-cultivated seedlings for three days yielded 2% frequency of trans format ion.
  • Thumbnail Image
    ThesisItemOpen Access
    In Vitro regeneration and Genetic Transformation of Pigeon Pea [Cajanus Cajan (L.) Millsp.] Genotype GT-102
    (Biotechnology Department, N. M. college of Agriculture Navsari Agricultural University, 2012-05) Parek, Mithilkumar Jitendrakumar; Mahatma, M. K.
    Pigeon pea [Cajanus cajan (L.) Millsp.] is an important legume crop cultivated across several country; and ranks fifth in area among pulses. It is consumed as a major source of protein (2 I %) to the human population in many developing countries while in India it is the second important food legume contributing to 80% of the global production. The most important yield constraint on Pigeon pea is the lepidopteron pest, Helicoverpa armigera. Therefore, in present investigation Agrobacterium mediated genetic transformation of cry1Aa3 in Pigeon pea genotype GT-102 was achieved. It was prerequisite to develop an efficient in vitro regeneration system and therefore here it was developed a simple and efficient regeneration protocol from apical meristem from young seedlings of Pigeon pea. While during experiment it is also observed that transformation by multiplication of existing meristem might resulted in very low frequency and chimeric transgenic plants which are difficult to handle. So, further in vitro genetic transfOlmation protocol was developed by using embryo axes.
  • Thumbnail Image
    ThesisItemOpen Access
    Isolation and Molecular characterization of Phosphate Solublising bacteria From Rhizospheric Soil
    (Biotechnology Department, N. M. college of Agriculture Navsari Agricultural University, 2012-01) Singh, Lochan; Jha, Sanjay
    Seven phosphate solubiiising bacteria, belonging to Enterobacteriaceae family, capable of utilizing Xylose and rock phosphate as the sole carbon and phosphate source were isolated by enrichment culture method at 28°C in aerobic conditions. This PSB exhibited mineral phosphate solubilising (MPS) phenotype on xylose and glucose, which are substrates for enzyme glucose dehydrogenase (GDH), as carbon sources. The Phosphate solubilising capacity of isolates ranged from 385~M-547~M at pH 5.0 in 60-168 hours in Xylose supplemented Tris minimal rock phosphate medium. The isolates secreted 26-46mM formic acid and 25-35mM xylonic acid by utilizing xylose whereas in glucose medium 2-7mM formic acid and 24-40mM gluconic acid was produced. In all the isolates, GDH activity was found when grown on Glucose and Xylose, indicating that it is constitutive and could act on a wide range of aldose sugars. PFL enzyme activity in all xylose utilizing cells was higher, since PFL IS a marker of anaerobic conditions thus all the isolates were facultative anaerobes. Efficiency of the isolates for phosphorus solubilization in natural environments was further confirmed by pot experiments on mung bean plants.