[Attempting to enhance the efficiency of T-DNA insertional mutant application in rice].
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.
Zhang
Ya-Fang
YF
Key Laboratory of Crop Functional Genomics of Jiangsu Province, Yangzhou University, Yangzhou 225009, China. yfzhang@yzu.edu.cn
Pan
Cun-Hong
CH
Li
Ai-Hong
AH
Tang
Wen
W
Wu
Ru
R
Chen
Zong-Xiang
ZX
Xu
Ai-Xia
AX
Pan
Xue-Biao
XB
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
D004269:DNA, Bacterial; C036483:T-DNA