An efficient method for the production of transgenic plants of peanut (Arachis hypogaea L.) through Agrobacterium tumefaciens-mediated genetic transformation

Sharma, K K and Anjaiah, V (2000) An efficient method for the production of transgenic plants of peanut (Arachis hypogaea L.) through Agrobacterium tumefaciens-mediated genetic transformation. Plant Science, 159 (1). pp. 7-19. ISSN 0168-9452

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Abstract

Cotyledon explants from mature peanut seeds (Arachis hypogaea L.) were optimized to obtain adventitious shoot buds with high frequencies (\90%). Efficient transformation of these cotyledons by using Agrobacterium tumefaciens strain C58 carrying neomycin phosphotransferase II (nptII) and ß-glucuronidase (GUS; uidA), or coat protein gene of the Indian peanut clump virus (IPCVcp) and nptII on binary vectors (pBI121; pROKII:IPCVcp) led to the production of a large percentage (55%) of transgenic plants. Transformed individuals were obtained through selection on medium containing 125 mg l�1 kanamycin. A large number of independently transformed plants (over 75) were successfully transplanted to the glasshouse. Integration of the transgenes and stable genetic transformants in the progeny were assessed by PCR amplification of 700-bp fragment of nptII and 585-bp of IPCVcp genes, and Southern blot hybridizations in the T1 generation of transgenic plants. Analysis of 35 transgenic plants of T1 generation from the progeny of a single transformation event suggested the segregation of a single copy insert in a 3:1 Mendelian ratio. On an average, 120–150 days were required between the initiation of explant transformation and transfer of rooted plants to the greenhouse. The cotyledon regeneration system proved to be an excellent vehicle for the production of a large number of independently transformed peanut plants. Shoot formation was rapid and prolific, and a large proportion of these shoots developed into fertile plants. The method reported here provides new opportunities for the crop improvement of peanut via genetic transformation. © 2000 Elsevier Science Ireland Ltd. All rights reserved.

Item Type:	Article
Divisions:	UNSPECIFIED
CRP:	UNSPECIFIED
Uncontrolled Keywords:	Agrobacterium tumefaciens; Arachis hypogaea; Groundnut; Indian peanut clump virus; Shoot regeneration; Transgenic plants
Subjects:	Mandate crops > Groundnut
Depositing User:	Library ICRISAT
Date Deposited:	26 Sep 2011 04:03
Last Modified:	26 Sep 2011 04:05
URI:	http://oar.icrisat.org/id/eprint/1922
Official URL:	http://dx.doi.org/10.1016/S0168-9452(00)00294-6
Projects:	UNSPECIFIED
Funders:	OPEC, Asian Development Bank
Acknowledgement:	We greatfully acknowledge the technical assistance of M. Lavanya, D. Pandary and Jagan Mohan Reddy, and assistance of L. Vidyasagar with photography. Dr R. Ortiz, Dr S. Sivaramakrishnan and Dr N. Seetharama provided useful suggestions on the manuscript. We thank Dr Mike Mayo, Scottish Crops Research Institute, for providing plasmid pROKII:IPCVcp, and his constant encouragement. Part of the funding for this work was provided by grants from OPEC and Asian Development Bank.
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