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2013 - 201512
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Subject: Molecular Biology and Biotechnology ×

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Now showing 1 - 9 of 12
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    ThesisItemOpen Access
    Wine production from over ripe guava fruits using saccharomyces cerevisiae
    (JNKVV, 2013) Patil, Yogesh Kalyanrao; Rajput, L.P.S.
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    ThesisItemOpen Access
    Studies on effect of cytokinin in combination with auxins on micropropagation efficiency of pomegranate (Punica granatum L.)
    (JNKVV, 2013) Thombare, Tulshiram Devidas; Tiwari, S.
    Abstract Pomegranate (Punica granatum L.) of family Punicaceae a native to Persia has been associated with the most ancient civilization in the Middle East. The total area under pomegranate cultivation in India is 100,000 ha yielding 0.45 million tons of fruit per year. It is cultivated commercially only in Maharashtra in over 25,000 ha and in Madhya Pradesh over 10,000 ha. The demand for tissue cultured plantlets of commercial plants both in agriculture and horticulture as well as in social forestry is growing day by day. Yields of pomegranate are lowering down because of the traditional method of propagation which is very cumbersome and carry several diseases. But the use of tissue culture techniques for fruit tree propagation has increased significantly. The explants were subjected to surface sterilization by treatment involving HgCl2 (0.5 %). Reliable and reproducible protocol to get healthy plants from different juvenile explants like stem segment without meristematic buds, stem segment with axillary buds and nodal segment with lateral buds for pomegranate cv ‘Bhagava’ has been developed. Out of these three explants, multiplications of shoots were observed in nodal segment with lateral buds on MS medium fortified with 3 mg/l BAP. MS medium containing 0.5 mg/L of GA3 was found to be the best for shoot elongation. Elongated shoots were transferred in MS media fortified 0.1 to 0.5 mg/L of NAA and 0.1 to 0.5 mg/L of IBA for rhizogenesis but root initiation was not started within 25 days incubation period as it may require some more time. Present study is carried out with an aim to develop a protocol for commercial production of pomegranate. Although, root induction was not observed in present investigation, but the multiplication and proliferation of shoots showed potential of this micropropagation protocol to be used in large-scale disease free true to type clonal multiplication.
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    ThesisItemOpen Access
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    ThesisItemOpen Access
    Molecular diversity analysis of whitefly (Bemisia tabaci) collected from different regions of madhya pradesh
    (JNKVV, 2013) Kale, Sumit Prakashrao; Tiwari, S.
    ABSRACT The present study entitles “Molecular diversity analysis of whitefly (Bemisia tabaci) collected from different regions of Madhya Pradesh” was carried out with the objectives to standardize genomic DNA isolation method and analyze the genetic diversity of whitefly using DNA fingerprinting. This study aimed to employ DNA based method to clarify the existing of biotype among whitefly populations in East Madhya Pradesh, India. The genomic DNA of whitefly collected from different geographical regions of East Madhya Pradesh was isolated by standardize protocol (SDS + Proteinase K) produce intact band with high molecular weight. The mtCOI primer was used for identification of B biotype. Amplified products were resolved by electrophoresis on 1.2% agarose gel and photographed under gel documentation system. Specific mtCOI primer CI-J-2195 and L2-N-3014 showed the presence of ~880bp bands which confirmed the presence of B biotype in different geographical regions of East Madhya Pradesh. All identified B biotypes were used for molecular diversity analysis. The DNA samples were amplified with 5 RAPD primers in thermal cycler. Amplified products were resolved by electrophoresis on 1.2% agarose gel and photographed under gel documentation system. Four decamer primers amplified 8 RAPD marker loci. All 8 bands scored by RAPD markers found to be monomorphic. Average number of bands per primer was 2.00. One RAPD primer is not amplified in all population. No variation was found among whitefly samples collected from different geographical regions of East Madhya Pradesh. Its seems that B. tabaci found in there region of collection are of conserved type.
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    ThesisItemOpen Access
    Studies on Agrobacterium-Mediated Transformation in Oat (Avena sativa L.)
    (JNKVV, 2013) Dattgonde, Nagesh Raosaheb; Tiwari, S.
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    ThesisItemOpen Access
    Amplification of Phy Gene From Bacillus Species sp. Isolated from Soil
    (JNKVV, 2013) Shinde, Vishwajeet; Tantwai, Keerti
    ABSTRACT Phytases (myo-inositol hexakisphosphate phosphohydrolase), also known as phytase-degrading enzymes, hydrolyses phytate to myo-inositol and phosphoric acid in stepwise manner forming myo-inositol phosphate intermediates. Phytases are widespread in nature because they can be found in animals, plants and microorganisms. Phytases produced biotechnologically have emerged as indispensable entities in feed and food industry for increased bioavailability of essential minerals. In plant-derived foods, phytate acts as an antinutritive factor by causing mineral deficiency due to efficient chelation of metal ions and by forming complexes with proteins. Unlike ruminates, monogastric animals such as pigs and poultry lack phytase-producing microorganisms in their gut. The addition of microbial phytase to the feedstuff for monogastric animals can substantially improve phosphorus utilization, reduce excretion of phosphorus in the faces and counteract the antinutritional properties of phytase. Ongoing interest in isolation of new bacterial strains producing phytase and optimization of this enzyme produces the recombinant feed poses the need for further investigation. The present investigation, entitled “Amplification of phy gene from Bacillus spp. isolated from soil” deals with the amplification and sequencing of phy gene with the help of phy gene specific primers. Genomic DNA was isolated from the single colonies of all the four bacterial isolates (BS 1, BS 2, BS 3 and BS 4). Then the purified DNA was used for the PCR amplification at 58ºC annealing temperature by using phy gene specific primers. The amplified PCR product showed fragment of ~650 bp in size. The amplified sequences of phy gene were sequenced and sequencing revealed the length of 634 and 635 bp from BS2 and BS4 isolates respectively. In silico analysis was carried out by using different software’s which showed 69.7% homology with B. subtilis strains. Phylogenetic tree also justified similarities with other Bacillus spp. Analysis of ORF and amino acid sequence was carried out, the nucleotide sequences showed the complete open reading frame and deduced amino acid sequences showed two sites of stop codon.
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    ThesisItemOpen Access
    Molecular Screening of Soybean Germplasm Resistant to Rhizoctonia Root Rot
    (JNKVV, 2013) Yeole, Pankaj Tryambak; Tantwai, Keerti
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    ThesisItemOpen Access
    Studies on in vitroregeneration of water chestnut (Trapa natans)
    (JNKVV, 2014) Sawalakhe, Roshani; Tiwari, S
    Abstract The water chestnut (Trapa natans), commonly known as ‘singhara’ in India, belongs to the monogeneric family trapaceae. The water chestnut is an annual hydrophyte, which bears starchy edible fruits and is grown in the shallow freshwater basins of tropical, subtropical and temperate zones. It is a rooted aquatic herb with rosettes of trullate (trowel-shaped) floating leaves. Water chestnut has been considered a minor crop because of its low yield and lack of suitable cultivation techniques. Indeed, there is a paucity of information on procedures to improve its cultivation and performance. Micropropagation has become an important aspect of commercial nursery propagation of many plants, especially of ornamentals, with possible extension to crops such as water chestnut. Regeneration is a prerequisite for the application of modern methods of in vitro culture for crop improvement. During present investigation, in vitro regeneration in water chestnut using different explants have been studied. Experiments were conducted to select most responding explant of water chestnut and growth regulator fortified in MS culture medium for efficient plant regeneration. For the in vitro regeneration studies in water chestnut from different explants viz. mature embryo, cotyledonary node and nodal cuttings cultured on MS medium supplemented with different concentrations and combinations of growth regulators. Explants collected from the field were surface sterilized with HgCl2, Tween-20, 70% ethanol followed by HgCl2. Culture initiation from explants was achieved on MS medium with BAP (1-5 mg/l), 2,4-D (1-5 mg/l). For seed germination or initiation of culture from mature embryo showed the best results on medium fortified with 2 mg/l 2,4-D (74%) germination whereas, lowest germination frequencies (28%) were observed in on combination of 5 mg/l 2,4-D which is near about the control (30%). Callus induction with mature embryo was observed within 25-30 days from mature embryos, being maximum (68%) on MS medium containing 2 mg/l 2,4-D alone and these callus were used as a explants for culture initiation by different explants. MS medium and B5 medium were also tested in which MS medium fortified with 3 mg/l 2, 4-D (74%) showed best response to explants in medium. The lowest response was observed on B5 media fortified with 3 mg/l BAP (45%). MS media responded better than B5 media. Maximum shoot initiation and multiplication was observed with in vitro germinated explants i.e cotyledonary node explants on MS medium containing 3 mg/l BAP (74%) and lowest with MS medium containing 5 mg/l BAP (28%) which is near about the control (30%). MS medium with combination of BAP and NAA was found suitable for multiple shoot formation. 3-4 in numbers of multiple shoots were obtained from each explant. The shoots regenerated in initiation experiment were elongated on MS medium with various concentration of GA3 and 0.5 mg/l of GA3 showed best elongation and proliferation of shoots. Elongated shoots were transferred in different concentration of auxins (IBA) for root induction. Up to the 24-28 days root formation do not occurs. Germination frequency of water chestnut seed is low, with a maximum of only about 60% germinated in the growing period. Efficient production of water chestnut in paddy fields requires mass production of uniform propagules and high-frequency seed germination. Therefore tissue culture is a suitable method to mass propagate aquatic plant species and offers several advantages for industry. Among the advantages is good quality of planting materials which are disease and the results of the present study clearly led to the conclusion that different explants, mature embryo of water chestnut may be preferred over other explants. It was observed with initiation experiments that media fortified with BAP and 2,4-D exhibited significant effect on seed germination as well as in case of 2,4-D, it induces callus formation when, supplied in medium. MS medium fortified with the BAP and NAA was shows better results on shoot induction from cotyledonary node. Shoot elongation and proliferation was obtained with GA3. From the experiments on rhizogenesis MS medium fortified with IBA in different concentration was used but root initiation was not observed within 25 days incubation periodvirus free at a competitive price while conserving aquatic plants in their natural habitat. During the present study, efforts have been made to develop efficient regeneration protocol for locally adapted cultivars using different explants, with consideration of urgency to use transgenic technologies for improvement of water chestnut. For the in vitro regeneration studies in water chestnut from different explants viz. mature embryo, cotyledonary node and nodal cuttings cultured on MS medium supplemented with different concentrations and combinations of growth regulators.
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    ThesisItemOpen Access