IN-PLANTA TRANSFORMATION OF SOYBEAN (Glycine max L.) TO ENHANCE SALINITY AND DROUGHT TOLERANCE
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Date
2017-10-10
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Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra.
Abstract
In the present investigation entitled “In-Planta transformation of soybean (Glycine max L.) to enhance salinity and drought tolerance” was under taken with the view to develop In-planta regeneration system for elite soybean genotype. The study was carried out at Biotechnology Centre, Dr. PDKV, Akola during year 2015-16.
In vitro culture and genetic transformation of soybean is difficult due to its recalcitrant nature. Establishment of gene transfer procedure is prerequisite to devolved transgenic plant of soybean in a shorter period. Therefore, the transformation was perform using the most popular soybean cv., JS-335 to optimize the factor influencing transformation efficiency through Agrobacterium tumefaciens based in-planta transformation using LBA4404 strain harboring gene, PDH45. In the present study four strategies were employed for in-planta transformation i.e. infection to seed, infection to seed and then incubation for 24 h in Agrobacterium, infection to every developing stage and infection to flowering and pod filling stage. Several parameters, such as optical density, incubation time, acetosyringone concentration influencing In-planta transformation, have been evaluated in this study. In seed radicle treatment, number of flower and pod formation was highest among the treatments. All the plant samples (T1) tested were found to be negative after PCR confirmation with gene specific primer.
Similarly, in treatment injected to seed radicle and incubated for 30 min. out of 50 sample two samples were found to be positive after PCR analysis with gene specific primer.
Putative transgenics were screened by PCR confirmation. It helped to eliminate non-transgenic. The method also decreased the time and effort involved in the screening process The procured plasmid containing PDH 45 gene was transformed into the Agrobacterium using In-planta method without using helper strain and confirmed by restriction digestion and the gene specific set of primers.
Description
In-planta transformation provide opportunity in order to develop reliable and rapid transgenic system. In-planta transformation does not required in-vitro regeneration and cannot pass through tissue culture cycles that means a non tissue culture based method of generating transgenic soybean.
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Citation
LUNGE AKSHAY RAJENDRA. (2017). In-Planta Transformation of soybean (Glycine max L.) to enhance salinity and drought tolerance. Biotechnology Centre, Department of Agricultural Botany. Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola. M. Sc. 2017. Print. xii, 65p. (Unpublished).