Research Project
Grant-in-Aid for Scientific Research (C)
We constructed T-DNA insertional lines of a model legume, Lotus japonicus, using a multifunctional vector for gene and exon activation tagging. The vector had the CaMV 35S promoter together with two additional enhancer elements, the start codon, and splice donor and acceptor sites facing to the left border, in anticipation of the activation of T-DNA flanking genes and forced expression of flanking exons. The improved transformation technique yielded more than 3,500 lines, including 45 dominant mutant candidates with abnormal phenotypes in aerial parts, nodules and roots. Among selected 44 lines, one copy of T-DNA was inserted into the genome of 37 lines (84%). The T-DNA flanking regions of seven lines were isolated by thermal asymmetric interlaced(TAIL)-PCR or reverse transcription(RT)-PCR, and the corresponding genomic clones were analyzed. The transcripts of four genes adjacent to T-DNA out of eleven genes tested were increased in the T_1 generation, demonstrating that gene and exon activation effects by the newly developed tagging vector are heritable. This is the first report on the activation tagging of a legume, and the T-DNA insertional population of L.japonicus will provide legume-specific dominant mutants.
All 2005 2004 2003 2002 Other
All Journal Article (15 results) Book (1 results) Publications (1 results)
Plant Physiology 137
Pages: 882-891
Biochemical and Biophysical Research Communications 330
Pages: 907-913
Plant Physiology 137(3)
Biochemical and Biophysical Research Communications 330(3)
DNA Research 11
Pages: 263-274
DNA Research 11(4)
Plant and Cell Physiology 44
Pages: 103-112
10010272519
Plant Physiology 131
Pages: 941-951
Plant and Cell Physiology 44(2)
Plant Physiology 131(3)
Plant Cell Reports 21
Pages: 238-243
Plant Journal 31
Pages: 555-564
Plant Journal 31(5)
Plant Cell Reports 21(3)
