About 25,000 rice T-DNA insertional mutant lines were generated using the vector pCAS04 which has both promoter-trapping and activation-tagging function. Southern blot analysis revealed that about 40% of these mutants...About 25,000 rice T-DNA insertional mutant lines were generated using the vector pCAS04 which has both promoter-trapping and activation-tagging function. Southern blot analysis revealed that about 40% of these mutants were single copy integration and the average T-DNA insertion number was 2.28. By extensive phenotyping in the field, quite a number of agronomically important mutants were obtained. Histochemical GUS assay with 4,310 primary mutants revealed that the GUS-staining frequency was higher than that of the previous reports in various tissues and especially high in flowers. The T-DNA flanking sequences of some mutants were isolated and the T-DNA insertion sites were mapped to the rice genome. The flanking sequence analysis demonstrated the different integration pattern of the right border and left border into rice genome. Compared with Arabidopsis and poplar, it is much varied in the T-DNA border junctions in rice.展开更多
With the completion of the Populus genome sequence in 2006,the study on functional genomics in Populus has become a major task.Establishment of Populus mutant population is an essential approach for forestry functiona...With the completion of the Populus genome sequence in 2006,the study on functional genomics in Populus has become a major task.Establishment of Populus mutant population is an essential approach for forestry functional genomics study.The activation tagging is a promising and powerful method to construct mutant library of Populus and to further discover new gene and elucidate its function now.A novel T-DNA activation tagging pCAS05 was constructed for forest functional genomics in this study.We could select conveniently and easily transgenic plants by spraying Basta because bar gene is constructed in this vector as a selection marker.Moreover,we could also construct a large scale mutant library in Populus by this activation tagging because of effectively selection of bar gene.展开更多
基金supported by the National High Technology Research and Development Program of China (No.2002AAZ2001)the National Natural Sciences Foundation of China (No.30270758 and 30621001)
文摘About 25,000 rice T-DNA insertional mutant lines were generated using the vector pCAS04 which has both promoter-trapping and activation-tagging function. Southern blot analysis revealed that about 40% of these mutants were single copy integration and the average T-DNA insertion number was 2.28. By extensive phenotyping in the field, quite a number of agronomically important mutants were obtained. Histochemical GUS assay with 4,310 primary mutants revealed that the GUS-staining frequency was higher than that of the previous reports in various tissues and especially high in flowers. The T-DNA flanking sequences of some mutants were isolated and the T-DNA insertion sites were mapped to the rice genome. The flanking sequence analysis demonstrated the different integration pattern of the right border and left border into rice genome. Compared with Arabidopsis and poplar, it is much varied in the T-DNA border junctions in rice.
文摘With the completion of the Populus genome sequence in 2006,the study on functional genomics in Populus has become a major task.Establishment of Populus mutant population is an essential approach for forestry functional genomics study.The activation tagging is a promising and powerful method to construct mutant library of Populus and to further discover new gene and elucidate its function now.A novel T-DNA activation tagging pCAS05 was constructed for forest functional genomics in this study.We could select conveniently and easily transgenic plants by spraying Basta because bar gene is constructed in this vector as a selection marker.Moreover,we could also construct a large scale mutant library in Populus by this activation tagging because of effectively selection of bar gene.
