Plant and Cell Physiology, 2000, Vol. 41, No. 7 881-888
© 2000 Oxford University Press
Antisense Waxy Genes with Highly Active Promoters Effectively Suppress Waxy Gene Expression in Transgenic Rice
1 Plantech Research Institute, 1000 Kamoshida, Aoba-ku, Yokohama, 227-0033 Japan 2 Laboratory of Plant Molecular Genetics, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, 630-0101 Japan 3 Department of Genetics and Botany, University of Georgia, Athens, GA 30602, U.S.A. 4 Division of Gene Expression and Regulation, National Institute for Basic Biology, Myodaiji, Okazaki, 444-8585 Japan
To regulate Waxy (Wx) gene expression by introducing antisense genes, we connected the 2.3 kb Wx cDNA having 450 bp of the Wx first intron in reverse orientation to rice Wx and maize alcohol dehydrogenase1 (Adh1) promoters and used these constructs to transform rice plants. Of 10 independent transgenic lines analysed, four lines showed various degrees of reduction in amylose and WAXY (WX) protein levels in the endosperm. In two transgenic lines, complete absence of amylose was observed which made the seeds opaque white like glutinous rice (amylose-deficient waxy (wx) mutant). In one of the transgenic lines, A1 line, the presence of the antisense Wx gene cosegregated with reduction of amylose content in the endosperm. In the same line, a reduction in the level of endogenous Wx mRNA was observed in immature endosperm. Interestingly, this reduction was observed only with mature spliced transcripts but not with unspliced transcripts. Reduced amylose synthesis was also observed in pollen grains of four transgenic lines. These results suggest that integrated antisense Wx gene caused a reduction in amylose synthesis in endosperms and pollen grains of transgenic rice carrying the antisense Wx cDNA. These results indicate that manipulation of starch and other carbohydrates in rice grain is possible using antisense genes.
5 Present address: Department of Biochemistry, University of Minnesota, 435 Delaware St S.E., Minneapolis, MN 55455, U.S.A.
6 Corresponding author: E-mail, simamoto@bs.aist-nara.ac.jp; Fax, +81-743-72-5509.
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