Biolistic-mediated genetic transformation of cowpea (Vigna unguiculata) and stable Mendelian inheritance of transgenes

We describe a novel system of exploiting the biolistic process to generate stable transgenic cowpea (Vigna unguiculata) plants. The system is based on combining the use of the herbicide imazapyr to select transformed meristematic cells after physical introduction of the mutated ahas gene (coding for...

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Bibliographic Details
Published in:	Plant cell reports Vol. 27; no. 9; pp. 1475 - 1483
Main Authors:	Ivo, Nayche L, Nascimento, Cristina P, Vieira, Lívia S, Campos, Francisco A. P, Aragão, Francisco J. L
Format:	Journal Article
Language:	English
Published:	Berlin/Heidelberg Berlin/Heidelberg : Springer-Verlag 01-09-2008 Springer-Verlag Springer Springer Nature B.V
Subjects:	Acetolactate Synthase - genetics Biolistics Biological and medical sciences Biomedical and Life Sciences Biotechnology Cell Biology DNA, Plant - genetics Fabaceae - drug effects Fabaceae - genetics Fundamental and applied biological sciences. Psychology Genes, Reporter Genetic engineering Genetic technics Genetic Transformation and Hybridization Imidazoles - pharmacology Inheritance Patterns Life Sciences Methods. Procedures. Technologies Microscopy, Electron Niacin - analogs & derivatives Niacin - pharmacology Plant Biochemistry Plant Sciences Plants, Genetically Modified - drug effects Plants, Genetically Modified - genetics Tissue Culture Techniques Transformation, Genetic Transgenes Transgenic animals and transgenic plants Transgenic plants Transgenic plant Gene transfer Gene expression Imazapyr Transgene Microprojectile bombardment Genetic transformation Gene expression . Gene transfer Vigna unguiculata Genetic transfer Leguminosae Dicotyledones Angiospermae Spermatophyta Inheritance(genetics) Transgenesis Transgene . Transgenic plant
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Summary:	We describe a novel system of exploiting the biolistic process to generate stable transgenic cowpea (Vigna unguiculata) plants. The system is based on combining the use of the herbicide imazapyr to select transformed meristematic cells after physical introduction of the mutated ahas gene (coding for a mutated acetohydroxyacid synthase, under control of the ahas 5' regulatory sequence) and a simple tissue culture protocol. The gus gene (under control of the act2 promoter) was used as a reporter gene. The transformation frequency (defined as the total number of putative transgenic plants divided by the total number of embryonic axes bombarded) was 0.90%. Southern analyses showed the presence of both ahas and gus expression cassettes in all primary transgenic plants, and demonstrated one to three integrated copies of the transgenes into the genome. The progenies (first and second generations) of all self-fertilized transgenic lines revealed the presence of the transgenes (gus and ahas) co-segregated in a Mendelian fashion. Western blot analysis revealed that the GUS protein expressed in the transgenic plants had the same mass and isoelectric point as the bacterial native protein. This is the first report of biolistic-mediated cowpea transformation in which fertile transgenic plants transferred the foreign genes to next generations following Mendelian laws.
Bibliography:	http://dx.doi.org/10.1007/s00299-008-0573-2 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0721-7714 1432-203X
DOI:	10.1007/s00299-008-0573-2