Regeneration of transgenic plants expressing the GFP gene from rape cotyledonary and leaf explants: Effects of the genotype and ABA

Regeneration of transgenic plants expressing the GFP gene from rape cotyledonary and leaf... Cotyledons of five-day-old seedlings and leaves of 6-week-old plants of two rape cultivars (Brassica napus L., cvs. Westar and Podmoskovnyi) were co-cultured with the culture of Agrobacterium tumefaciens cells comprising the genetic construct with the marker gfp gene, on Murashige and Skoog nutrient medium supplemented with benzyladenine, NAA, and ABA in various combinations. A capacity for regeneration on both types of explants was rather high, but leaf explants produced weakly differentiated shoots and most of them were vitrificated. On cotyledonary explants of transformed rape plants of both cultivars expressing the gfp gene, regeneration frequency was 70%. On leaf explants, it was much lower (47% in cv. Westar and 28% in cv. Podmoskovnyi). The gfp gene was expressed on all stages of shoot development. On primary, starting differentiation calli, we observed the strongest fluorescence of GFP in meristematic and vascular tissues. On leaf blades, GFP fluorescence was much brighter in old than young leaves; often it was observed only in the cell groups; it. PCR analysis of seed generation of transformants showed that some plants did not follow the Mendelian inheritance of a monogenic trait (transgene) in self-pollinated plants. This phenomenon could be explained as a result of meiotic recombination or production of genotypic chimeric organisms at regeneration. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Russian Journal of Plant Physiology Springer Journals

Regeneration of transgenic plants expressing the GFP gene from rape cotyledonary and leaf explants: Effects of the genotype and ABA

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Publisher
SP MAIK Nauka/Interperiodica
Copyright
Copyright © 2012 by Pleiades Publishing, Ltd.
Subject
Life Sciences; Plant Physiology; Plant Sciences
ISSN
1021-4437
eISSN
1608-3407
D.O.I.
10.1134/S1021443712030089
Publisher site
See Article on Publisher Site

Abstract

Cotyledons of five-day-old seedlings and leaves of 6-week-old plants of two rape cultivars (Brassica napus L., cvs. Westar and Podmoskovnyi) were co-cultured with the culture of Agrobacterium tumefaciens cells comprising the genetic construct with the marker gfp gene, on Murashige and Skoog nutrient medium supplemented with benzyladenine, NAA, and ABA in various combinations. A capacity for regeneration on both types of explants was rather high, but leaf explants produced weakly differentiated shoots and most of them were vitrificated. On cotyledonary explants of transformed rape plants of both cultivars expressing the gfp gene, regeneration frequency was 70%. On leaf explants, it was much lower (47% in cv. Westar and 28% in cv. Podmoskovnyi). The gfp gene was expressed on all stages of shoot development. On primary, starting differentiation calli, we observed the strongest fluorescence of GFP in meristematic and vascular tissues. On leaf blades, GFP fluorescence was much brighter in old than young leaves; often it was observed only in the cell groups; it. PCR analysis of seed generation of transformants showed that some plants did not follow the Mendelian inheritance of a monogenic trait (transgene) in self-pollinated plants. This phenomenon could be explained as a result of meiotic recombination or production of genotypic chimeric organisms at regeneration.

Journal

Russian Journal of Plant PhysiologySpringer Journals

Published: Apr 15, 2012

References

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