OPTIMIZATION OF PRETREATMENTS FOR ENHANCING Agrobacterium MEDIATED TRANSFORMATION EFFICIENCY IN RICE VAR. PKV-HMT.

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Date
2017-08-28
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Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola
Abstract
The present investigation entitled “Optimization of pretreatments for enhancing Agrobacterium mediated transformation efficiency in rice var. PKV-HMT” was carried out with a view to develop resistance against yellow stem borer. Rice (Oryza sativa L.) is the important staple food for the world's human population. Several biotic and abiotic factors affect the rice production in India. However, losses due to yellow stem borer are huge and almost ubiquitous though out India and region. The estimated loss from stem borer damage varies from 3-95%. The present investigation aims to develop an efficient protocol for in vitro regeneration and genetic transformation of Indica rice cultivar viz. ‘PKV-HMT’. The Agrobacterium strain EHA105 harbouring vector pBinAR containing cry1Aabc gene was used for the transformation of rice cultivar PKV-HMT. The mature dehusked seeds of rice were surface sterilized with Bavistin (1%) for 15 minutes followed by 0.1% HgCl2 for 10 minutes which provides the maximum asceptic culture with better survival percentage. 1.25 mg/L 2,4-D in the MS medium induced profuse, white, friable, calli in rice variety ‘PKV-HMT’. Pretreatments to calli was found effective, comprises 6th day old callus along with 4 min heat shock to callus before infection and 7% maltose concentration in cocultivation medium showed higher transformation and regeneration frequency. MS medium supplemented with 3.0 mg/L BAP and 0.25 mg/L IAA induced the maximum multiple shoots and roots. Co-cultivation for 2 days at bacterial inoculums density (0.2) at 620 nm in presence of acetosyringone (100µM) increased the transformation efficiency. Selection medium containing 50 mg/L kanamycin and 600 mg/L cefotaxime was found to be the suitable for efficient screening of putative transformants in cv. ‘PKV HMT’ variety. This attempt of genetic transformation is a steping stone to develop transgenic PKV-HMT resistant against YSB.
Description
The present investigation entitled “Optimization of pretreatments for enhancing Agrobacterium mediated transformation efficiency in rice var. PKV-HMT” was carried out with a view to develop resistance against yellow stem borer.Pretreatments to calli was found effective, comprises 6th day old callus along with 4 min heat shock to callus before infection and 7% maltose concentration in cocultivation medium showed higher transformation and regeneration frequency. This attempt of genetic transformation is a steping stone to develop transgenic PKV-HMT resistant against YSB.
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Citation
MEHER, MISS. POOJA APPARAO. (2017).Optimization of pretreatments for enhancing agrobacterium mediated transformation efficiency In rice var. PKV-HMT. Biotechnology Centre. Department of Botany, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola. M.Sc. 2017. Print. xiii. 54p. (Unpublished).
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