浙江农林大学学报2024，Vol.41Issue(2)：314-321,8.DOI:10.11833/j.issn.2095-0756.20230304
滇杨组织培养再生及遗传转化体系建立
Regeneration system and genetic transformation of Populus yunnanensis
摘要
Abstract
[Objective]This study is aimed to establish a highly efficient system for regeneration and genetic transformation of in vitro leaves of Populus yunnanensis so as to help genetic engineering breeding research in P.yunnanensis.[Method]First,with P.yunnanensis leaves as explants,an investigation was conducted of the effects of different combinations of exogenous hormone concentrations on the induction of callus,adventitious bud differentiation and rooting so as to obtain regenerated seedlings of P.yunnanensis.Then,Agrobacterium tumefaciens mediated method was used to explore the effects of bacterial concentration,infection time and co-culture time on the genetic transformation efficiency of P.yunnanensis.[Result]The optimal medium for callus induction was 1/2 MS + 0.005 mg·L-1 thidiazuron(TDZ)+ 0.010 mg·L-1 1-naphthylacetic acid(NAA),with an induction rate of 91.7%.The optimal medium for the induction of adventitious bud was 1/2 MS + 0.002 mg·L-1 TDZ + 0.010 mg·L-1 NAA,with an induction rate of 75.0%whereas the optimal medium for rooting was 1/2 MS + 0.010 mg·L-1 NAA + 0.100 mg·L-1 3-indolebutyric acid(IBA),and the rooting rate was as high as 96.7%,with an average number of 2.57 roots.The optimal concentration of transforming bacterium D(600)was 0.2,the infection time was 5 min,and the co-cultivation time was 2 days.During differentiation,the optimal concentration of cefotaxime to inhibit the growth of A.tumefaciens was 200 mg·L-1,and the optimal concentration of kanamycin in the screening medium for resistant buds was 20 mg·L-1.The regenerated plants were identified by β-glucuronidase(GUS)staining and PCR,identified 20 positive plants,with a positive transformation rate of 45.45%.[Conclusion]The leaf regeneration system and genetic transformation system of P.yunnanensis were successfully established.[Ch,5 fig.5 tab.27 ref.]关键词
滇杨/叶片/愈伤组织/再生体系/遗传转化Key words
Populus yunnanensis/leaf/callus/regeneration system/genetic transformation分类
农业科技引用本文复制引用
张晓琳,纵丹,李嘉其,杨玲,余进德,何承忠..滇杨组织培养再生及遗传转化体系建立[J].浙江农林大学学报,2024,41(2):314-321,8.基金项目
国家自然科学基金资助项目(31860219) （31860219）
云南省教育厅自然科学基金资助项目(2021Y272) （2021Y272）
