The Acquisition of KdLEC1 and GbLEC1 Transgenic Cotton Regeneration Plantlets

Zhao Zhenjun;Zhang Xuejiao;Cui Baiming;Laboratory of Agricultural Biotechnology in College of Life Science of Shihezi University;

Xinhai15;;Xinluzao33;;LEC1;;Somatic embryogenesis;;Regeneration

In order to explore the effects of LEC1 on somatic embryogenesis of cotton, in this research, dexamethasone(DEX) inducible expression vector p INDEX3 was used to mediate respectively the truncated LEC1 of Kalanchoёdaigremontiana(Kd LEC1) on Xinhai15(Gossypium barbadense L.) hypocotyl and two LEC1 gene family of cotton(Gb LEC1 A,Gb LEC1B) and Kd LEC1 on Xinluzao33(Gossypium hirsutum L.) embryogenic callus by the agrobacterium to perform genetic transformation, somatic embryo induction as well as regeneration. Then,Gb LEC1 A, Gb LEC1 B and Kd LEC1 transgenic plants were obtained. Moreover, transgenic Xinhai15 embryogenic callus lines were treated respectively by DEX and no DEX suspension induction. The Kd LEC1 relative expression level was analyzed by real-time quantitative PCR. The results indicated that two Kd LEC1 transgenic Xinhai15 regeneration plants were obtained after regeneration for up to 13 months and PCR analysis. Besides, the study also obtained respectively one Gb LEC1 A, Gb LEC1 B and Kd LEC1 transgenic Xinluzao33 regeneration plants, which regeneration period was about 10 months. Real-time quantitative PCR analysis showed that Kd LEC1 relative expression level fluctuates between 2.5 and 23 times, demonstrating that p INDEX3 can induce exogenous gene Kd LEC1 expression at different levels. The study is aimed at providing fine materials for functional study of LEC1 in the process of cotton somatic embryogenesis and theoretical guidance for cotton germplasm innovation.