title: Transgenic cowpea (Vigna unguiculata) seeds expressing a bean alpha-amylase inhibitor 1 confer resistance to storage pests, bruchid beetles.
abstract: Cowpea is one of the important grain legumes. Storage pests, Callosobruchus maculatus and C. chinensis cause severe damage to the cowpea seeds during storage. We employ a highly efficient Agrobacterium-mediated cowpea transformation method for introduction of the bean (Phaseolus vulgaris) alpha-amylase inhibitor-1 (alphaAI-1) gene into a commercially important Indian cowpea cultivar, Pusa Komal and generated fertile transgenic plants. The use of constitutive expression of additional vir genes in resident pSB1 vector in Agrobacterium strain LBA4404, thiol compounds during cocultivation and a geneticin based selection system resulted in twofold increase in stable transformation frequency. Expression of alphaAI-1 gene under bean phytohemagglutinin promoter results in accumulation of alphaAI-1 in transgenic seeds. The transgenic protein was active as an inhibitor of porcine alpha-amylase in vitro. Transgenic cowpeas expressing alphaAI-1 strongly inhibited the development of C. maculatus and C. chinensis in insect bioassays.
authors.affiliation: Department of Biotechnology, Indian Institute of Technology Guwahati, Guwahati, Assam 781039, India.
authors.forename: Siva Kumar
authors.initials: SK
authors.lastname: Solleti
authors.forename: Souvika
authors.initials: S
authors.lastname: Bakshi
authors.forename: Jubilee
authors.initials: J
authors.lastname: Purkayastha
authors.forename: Sanjib Kumar
authors.initials: SK
authors.lastname: Panda
authors.forename: Lingaraj
authors.initials: L
authors.lastname: Sahoo
mesh_terms: D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D007887:Fabaceae / Q000235:genetics* / Q000378:metabolism / Q000469:parasitology*; D015870:Gene Expression; D017343:Genes, Plant; D005822:Genetic Vectors; D010935:Plant Diseases / Q000469:parasitology; D037121:Plant Lectins / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified; D012231:Rhizobium / Q000235:genetics; D012639:Seeds / Q000378:metabolism / Q000469:parasitology; D048889:Weevils / Q000472:pathogenicity*
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D018744:DNA, Plant; D037121:Plant Lectins; C036483:T-DNA; C061510:alpha-amylase inhibitor, Phaseolus vulgaris
references.citation: Transgenic Res. 2008 Jun;17(3):425-36
references.pmid: 17638109
references.citation: Plant Cell Rep. 2006 Apr;25(4):304-12
references.pmid: 16244884
references.citation: Proc Natl Acad Sci U S A. 2000 Apr 11;97(8):3820-5
references.pmid: 10759552
references.citation: Plant J. 1994 Aug;6(2):271-82
references.pmid: 7920717
references.citation: Plant Physiol. 1995 Apr;107(4):1233-1239
references.pmid: 12228429
references.citation: J Econ Entomol. 2007 Aug;100(4):1416-22
references.pmid: 17849896
references.citation: J Biosci. 2006 Sep;31(3):339-45
references.pmid: 17006016
references.citation: Plant Physiol. 1990 Jun;93(2):805-10
references.pmid: 16667540
references.citation: Exp Cell Res. 1968 Apr;50(1):151-8
references.pmid: 5650857
references.citation: Anal Biochem. 1976 May 7;72:248-54
references.pmid: 942051
references.citation: Pest Manag Sci. 2007 May;63(5):440-6
references.pmid: 17340671
references.citation: Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7603-7
references.pmid: 8052627
references.citation: Plant Mol Biol. 2000 Jul;43(4):495-502
references.pmid: 11052201
references.citation: Nucleic Acids Res. 1984 Nov 26;12(22):8711-21
references.pmid: 6095209
references.citation: Biochim Biophys Acta. 1999 Jun 15;1432(1):104-12
references.pmid: 10366733
references.citation: Proc Natl Acad Sci U S A. 1974 Sep;71(9):3672-6
references.pmid: 4530328