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Subject: Plant Biochemistry × End date: 2009 × Start date: 2005 × Type: Thesis ×
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    ThesisItemOpen Access
    Cloning and Characterization of Serine Protease Inhibitor Genes (SPIs) From Plants
    (UAS, Dharwad, 2005) D.A.Shirani; Sumangala Bhat
    Plant serine protease inhibitors (SPIs) can be effective in developing transgenic crop plants with resistance to insect attack. Such genes have few problems related to codon bias, RNA processing and protein stability. In the present study, an attempt was made to clone serine protease inhibitor genes from soybean (Glycine max), cowpea (Vigna unguiculata) and yard long bean (Vigna sesquipedalis). Using specific primers, genes encoding kunitz trypsin inhibitor of soybean (SKTI) and cowpea trypsin inhibitor (CpTI) from cowpea and yard long bean were amplified and cloned into pTZ57R/T vector. The clones were confirmed through PCR amplification and restriction digestion analysis. Further, the clones were sequenced and analyzed in silico. Cloned kunitz trypsin inhibitor of soybean showed 99 and 100 per cent homology with reported kunitz inhibitor genes of soybean at nucleotide and protein level respectively. The cloned CpTI of cowpea and yard long bean showed 99% homology with Phaseolus vulgaris trypsin proteinase inhibitor gene and published CpTI coding sequences (324bp). The deduced amino acid sequence of cloned cowpea CpTI showed 100% homology with Phaseolus vulgaris trypsin inhibitor whereas CpTI of yard long bean showed 98 percent homology. Further, to facilitate transformation of crop plants, soybean kunitz trypsin inhibitor and CpTI of cowpea and yard long bean were sub cloned into a plant transformation vector pHS100 at BamH1 and Xba1 restriction sites. The recombinant clones pHS-SBC1, pHS-CpC1 and pHS-YLBC3 were mobilized into Agrobacterium tumefaciens LBA4404 by triparental mating. The A. tumefaciens with recombinant clones pHS-SBC1 and pHS-CpC1 were used to transform tobacco and the presence of inserts in kanamycin resistant plants was checked through PCR with specific primers. The events have to be confirmed further and expression tested.
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    ThesisItemOpen Access
    Yield and quality of grapes (cv. thompson seedless) in relation to soil fertility status of vineyards in Bijapur taluk of karnataka
    (UAS, Dharwad, 2005) H.Yogeeshappa; S.I.Tolanur
    Nutrient depletion has become more pronounced consequent to other exploitation of natural resources to meet the food requirement in view of an ever-increasing population. It is therefore important to examine the fertility status of soils from time to time. A systematic investigation was conducted on soil fertility status of the low and high yielding vineyards in Bijapur taluk. Sixty vineyards were surveyed during 2006-07. Out of 60, 30 vineyards were selected based on previous year yield data for the purpose of collecting soil and petiole samples. The vineyards which produced less than 10 tonnes per acre and those which produced more than 10 tonnes per acre were categorised as low yielding and high yielding vineyards, respectively. [ All the soils belong to clay in texture. The pH of soils was alkaline in reaction (8.19 – 8.50). The soils were non saline. The organic carbon content of the soils ranged from medium to high. The CEC of the soil ranged from 49.45 to 59.48 cmol (p+) kg-1. The available nitrogen content of the soils ranged from 164.20 to 231.71 kg ha-1. The available phosphorus content in the soils ranged from 18.64 to 35.89 kg ha-1. The available potassium content of the soil raged from 432.64 to 543.39 kg ha-1. The available sulphur content of the soils ranged from 22.81 to 35.32 kg ha-1. The soils were optimum in available micronutrients. The nutrients in the petiole were non-significant and positively correlated with yield except P and K at 45 days after April pruning. The average cane girth after April pruning (vigour of vine) was found more in case of high yielding vineyards than in low yielding vineyards. The average girth of fruiting shoot was also more in high yielding vineyards. The better management practices by the farmers ensure higher yields.