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

abstract

  • Cotyledon explants from mature peanut seeds (Arachis hypogaea L.) were optimized to obtain adventitious shoot buds withhigh frequencies (\90%). Efficient transformation of these cotyledons by using Agrobacterium tumefaciens strain C58 carryingneomycin 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 transgenicplants. Transformed individuals were obtained through selection on medium containing 125 mg l?1 kanamycin. A large numberof independently transformed plants (over 75) were successfully transplanted to the glasshouse. Integration of the transgenes andstable genetic transformants in the progeny were assessed by PCR amplification of 700-bp fragment of nptII and 585-bp ofIPCVcp genes, and Southern blot hybridizations in the T1 generation of transgenic plants. Analysis of 35 transgenic plants of T1generation from the progeny of a single transformation event suggested the segregation of a single copy insert in a 3:1 Mendelianratio. On an average, 120?150 days were required between the initiation of explant transformation and transfer of rooted plantsto the greenhouse. The cotyledon regeneration system proved to be an excellent vehicle for the production of a large number ofindependently transformed peanut plants. Shoot formation was rapid and prolific, and a large proportion of these shootsdeveloped into fertile plants. The method reported here provides new opportunities for the crop improvement of peanut viagenetic transformation. © 2000 Elsevier Science Ireland Ltd. All rights reserved

publication date

  • 2000
  • 2000