植物学报2025，Vol.60Issue(3)：425-434,10.DOI:10.11983/CBB24092
发根农杆菌介导的野葛毛状根遗传转化体系
Agrobacterium rhizogenes-mediated Transformation System of Pueraria lobata Hairy Roots
摘要
Abstract
INTRODUCTION:An efficient Agrobacterium rhizogenes-mediated transformation system for Pueraria lobata was estab-lished. RATIONALE:In this study,tissue-cultured plantlets of P.lobata were used as explants to investigate the effects of dif-ferent genotypes,A.rhizogenes strains,explants,precultivation times,infection times,culture days,subculture times,and culture methods on the efficiency of hairy root genetic transformation in P.lobata. RESULTS:The results indicated that the induction rate of hairy root formation was the highest when the immature leaves of YG-19 were used as the explant material,reaching 10.2％.A.rhizogenes K599 was identified as the most suitable strain.The optimal explant material was immature leaves that had just unfolded from the first to second nodes of the 5th to 13th generation tissue culture plantlets subcultured for 8 days.After 3 days of pre-culture and 15 minutes of bacterial infection,the highest induction rate of hairy roots reached 22.4％.The optimal type of culture medium for the proliferation of hairy roots in P.lobata was solid medium culture,and the fresh weight of hairy roots grown on solid medium was 75 times greater than that of hairy roots grown in liquid medium.PCR detection and fluorescence microscopy assays re-vealed that the expression of GFP and rolB genes in the hairy roots of P.lobata was stable,and the rate of cotransfor-mation was 80％. CONCLUSION:Genotype,A.rhizogenes strain,and culture duration were the most critical factors for the efficient genetic transformation of hairy roots in P.lobata.关键词
野葛/发根农杆菌/毛状根/遗传转化Key words
Pueraria lobata/Agrobacterium rhizogenes/hairy roots/genetic transformation引用本文复制引用
曾文丹,龙紫媛,李婕宇,严华兵,吴正丹,尚小红,曹升,陆柳英,肖亮,施平丽,程冬..发根农杆菌介导的野葛毛状根遗传转化体系[J].植物学报,2025,60(3):425-434,10.基金项目
广西自然科学基金(No.2023GXNSFBA026297)、广西科技重大专项(No.桂科AA23023035)、广西科技计划(No.桂科AB22080090)、广西薯类创新团队(No.nycytxgxcxtd-2023-11)和科技先锋队(No.桂农科盟202414) （No.2023GXNSFBA026297）
