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    Research HighlightItemOpen Access
    CHARACTERIZATION OF DRUDGERY OF WOMEN IN PRODUCTION SYSTEMS
    (PROFESSOR JAYASHANKAR TELANGANA AGRICULTURAL UNIVERSITY, RAJENDRANAGAR, HYDERABAD, 2016) Dr.A.MRUNALINI Technical Coordinator
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    ThesisItemOpen Access
    STANDARDIZATION OF AGROBACTERIUM MEDIATED TRANSFORMATION PROTOCOL IN TOMATO (Solanum lycopersicum L.) Cv. PKM-1.
    (ACHARYA N.G. RANGA AGRICULTURAL UNIVERSITY, RAJENDRANAGAR, HYDERABAD., 2008) HIMABINDU, K.B; SHANTI PRIYA, M
    The present investigation was carried out to standardize the Agrobacterium mediated transformation protocol in tomato and it is carried out at transgenic laboratory of Dept. of Plant Physiology, RARS, Tirupati and Dept. of Genetics and Plant Breeding, S.V. Agricultural College, Tirupati. Seeds treated with 5% NaOCl for 20 min and inoculated on MS medium with out sucrose and incubation in dark for three days produced healthy and uniform seedlings without contamination. Explants i.e. cotyledons and hypocotyls isolated from 10 day old seedlings were found ideal for high frequency of regeneration compared to younger or older seedlings. Among the two explants i.e. cotyledon and hypocotyls, cotyledons showed better response compared to hypocotyls. Hence cotyledonary explants from 10 days old in vitro seedlings of PKM-1 were used for further studies on regeneration and transgenic protocols. Among the various plant growth regulators combinations tried the best shoot regeneration was obtained when MS medium was supplemented with BAP 1.5 mg/L + Kinetin 1.0 mg/L and root regeneration was obtained when MS medium was supplemented with Kinetin 1.0 mg/L respectively. Cotyledonary explants excised out from 10 days old seedlings were incubated for 10 min with over night grown Agrobacterium culture and cocultivated for 2 days followed by transfer to media containing cefotaxime 500 mg/L for 4 days before transferring to the medium containing 75 mg/L kanamycin which was found to be optimum for checking the Agrobacterium growth. Higher plantlet survival (86%) was obtained in soilrite mixture and 9.6 days has been taken for acclimatization. The transformation was carried out using the Agrobacterium strain LBA4404 containing the binary vector pCAMBIA 2301 harboring npt II as selectable marker and GUS as reporter gene. Confirmation of the transgene integration in the putative transformants was done by using the histochemical staining and PCR. The transformation efficiency of 44.4% was obtained in the cultivar PKM-1. The transformation frequency was 3.5% and the GUS gene transient expression level in transformants was 44.4%. Thus, the present study successfully demonstrated the indirect regeneration of transgenic plants from cotyledonary explants through Agrobacterium mediated genetic transformation approach in tomato Cv. PKM-1. The standardized protocols of present study may be utilized for further transgenics development in PKM-1 cultivar genetic background. Seeds treated with 5% NaOCl for 20 min and inoculated on MS medium without sucrose and dark incubation for three days produced healthy and uniform seedlings without contamination. Cotyledons and hypocotyl explants isolated from 10 day old seedlings were found ideal for high frequency regeneration compared to younger or older seedlings. Comparatively cotyledons showed better explant response than hypocotyls. Hence, cotyledonary explants from 10 days old in vitro seedlings were used for further studies on regeneration and transgenic protocols. Best shoot and regeneration, was obtained when MS medium is supplemented with BAP 1.5 mg/L + Kinetin 1.0 mg/L and root regeneration, was obtained when MS medium is supplemented with Kinetin 1.0 mg/L respectively and higher plantlet survival (86%) was obtained in soilrite mixture with in 10 days. The transformation was carried out using the Agrobacterium strain LBA4404 containing the binary vector pCAMBIA 2301 harboring GUS as reporter gene. Cotyledonary explants excised out from 10 days old seedlings were incubated for 10 min with over night grown Agrobacterium culture and co-cultivated for 2 days followed by transfer to media containing cefotaxime 500 mg/L for 4 days before transferring to the medium containing 75 mg/L kanamycin was found to be optimum for checking the Agrobacterium growth. Stable integration of the transgene in the putative transformants was confirmed by using the histochemical staining and PCR assay.
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    Research HighlightItemOpen Access
    Extension Highlights 2001 - 02
    (Angrau Hyderabad, 2002)
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    Research HighlightItemOpen Access
    Advances In Mango Research
    (Fruit Research Station Sanagareddy Angrau) Vijaya N Et Al
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    Research HighlightItemOpen Access
    Extension Highlights 1991 - 92
    (Angrau Hyderabad, 1992)
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    Research HighlightItemOpen Access
    Package Of Practice In Horticulture
    (Angrau Hyderabad, 1999)
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    Research HighlightItemOpen Access
    Extension Highlights 2005 - 06
    (Angrau Hyderabad, 2006)
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    Research HighlightItemOpen Access
    Extension Highlights 2000 - 01
    (Angrau Hyderabad, 2001)
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    Research HighlightItemOpen Access
    Glimpses Of Recent Soil Science Research At Angrau
    (Angrau ; Hyderabad, 2004) Riazuddin Ahmed S Et Al