Identification Of Variation Traits And Characterization Of T-DNA Insertion Sites Of Different Transgenic Cotton Lines

Some trait variation or mutant individuals often appear in research of transgenic cottons. The reason is probably the inducement in the process of somatic culture or insertion mutagenesis caused by T-DNA integrating into cotton genome. Choosing a lot of transgenic lines from those variation lines, four transgenic lines among them has been compared with primitive receptor variety Jihe321(five near-isogenetic lines in theory ) and analyzed, and comparatively analyzed the difference between four transgenic lines and receptor variety Jihe321 in agricultural traits, quality traits. Four crosses obtained from four female parent transgenic lines and Jihe321 male parent. Four F₂ populations was identified the resistance to kanamycin, which 150 cotton plants were detected in the field, respectively. PCR amplification using primers of report gene NPTII was conducted to confirm the segregation ratio of negative plants to positive plants. Difference of quality traits and agronomy traits were comparied between PCR positive group and negative group in F₂ and F₃. Southern blotting was analyzed to a lot of transgenic lines. Meanwhile, Using TAIL-PCR (Thermal Asymmetric Interlace PCR) technology obtained and analyzed Transferred-DNA flanking sequences. Its result is as follows:1. Among four F₂ populations, the segregation ratio of positive plants to negative plants is both consistent with 3:1 by kanamycin and PCR detection. Southern blotting indicated that single copy inserted into the genome of four transgenic lines, respectively; Counting from one copy to four copies in other several transgenic lines. The result is better that HindIII enzyme cuts the cotton genome of the difference transgenic lines than other enzymes.2. F₂ populations and F_2:3 populations had been investigated quality, agricultural traits amounting to 14 traits indexes. According to PCR positive and PCR negative, each population was been divided into two groups, and compared the traits difference and dependence of T-DNA and traits between two groups. Among them...