title: [Attempting to enhance the efficiency of T-DNA insertional mutant application in rice].
abstract: T-DNA tagging method is a high throughput system for identifying and cloning novel genes from T-DNA-inserted mutant population created via genetic transformation by Agrobacterium tumefaciens. However, the efficiency of using T-DNA-inserted mutant population to clone genes in rice was much lower than in Arabidopsis. In this study, a rice tagged line with two copies of T-DNA segments inserted independently to each other was screened out via a series of verification tests, including the co-segregated analysis between the mutated character and the sequence of T-DNA or the genomic sequence flanking inserted T-DNA. From this tagged line, two inserted incidents were separated from the progeny population of a plant heterozygous in two tagged sites, and some plants with the target trait and one of the inserted incidents were obtained, which were important basic materials for the subsequently co-segregated analysis between the mutated character and the sequence of inserted T-DNA, and for cloning the mutant gene in future. Based on this study, we have some thoughts about the gene cloning from the T-DNA tagged lines with more than one inserted sequence independently and put forward to discuss with colleagues.
authors.affiliation: Key Laboratory of Crop Functional Genomics of Jiangsu Province, Yangzhou University, Yangzhou 225009, China. yfzhang@yzu.edu.cn
authors.forename: Ya-Fang
authors.initials: YF
authors.lastname: Zhang
authors.forename: Cun-Hong
authors.initials: CH
authors.lastname: Pan
authors.forename: Ai-Hong
authors.initials: AH
authors.lastname: Li
authors.forename: Wen
authors.initials: W
authors.lastname: Tang
authors.forename: Ru
authors.initials: R
authors.lastname: Wu
authors.forename: Zong-Xiang
authors.initials: ZX
authors.lastname: Chen
authors.forename: Ai-Xia
authors.initials: AX
authors.lastname: Xu
authors.forename: Xue-Biao
authors.initials: XB
authors.lastname: Pan
mesh_terms: D004269:DNA, Bacterial / Q000235:genetics*; D016254:Mutagenesis, Insertional; D012275:Oryza / Q000235:genetics* / Q000254:growth & development; D010641:Phenotype; D030821:Plants, Genetically Modified / Q000235:genetics / Q000254:growth & development; D014170:Transformation, Genetic
chemical_list: D004269:DNA, Bacterial; C036483:T-DNA