摘要
以大花美人蕉(Canna×generalis)无菌苗叶鞘为外植体,采用均匀设计与正交设计法,探讨美人蕉愈伤组织诱导与增殖、不定芽分化与增殖、生根培养和驯化移栽的影响因素,成功建立大花美人蕉叶鞘不定芽再生体系。结果表明,诱导愈伤组织的最佳培养基为MS+6.62 mg·L^(-1)6-苄基腺嘌呤(6-BA)+1.5mg·L^(-1)萘乙酸(NAA)+0.2 mg·L^(-1)噻苯隆(TDZ),诱导率为80.77%。暗培养14 d为愈伤组织增殖最适培养条件,增殖的最佳培养基为MS+2.3 mg·L^(-1)6-BA+2.28 mg·L^(-1)TDZ+0.1 mg·L^(-1)3-吲哚乙酸(IAA),增殖率为97.50%,增殖系数为4.28。不定芽分化的最佳培养基为MS+1.0 mg·L^(-1)6-BA+0.1 mg·L^(-1)IAA,分化率达88.00%。不定芽增殖的最佳培养基为MS+6 mg·L^(-1)6-BA+1.5 mg·L^(-1)NAA+0.2 mg·L^(-1)TDZ,增殖系数可达18.00。无菌苗在1/2MS+0.5 mg·L^(-1)NAA+30 g·L^(-1)蔗糖生根培养基上生根率为100%,移栽在体积比1:1:1的草炭-有机肥-土混合营养基质中成活率高达98.33%。本研究建立的大花美人蕉再生体系可为研究其种质资源保存、遗传转化体系建立、基因工程育种及新品种选育等提供技术支持。
The leaf sheaths in sterile seedlings of Canna×generalis were used as explants to investigate the factors influencing callus induction and proliferation,adventitious shoots differentiation and proliferation,rooting culture,and domestication transplanting using uniform and orthogonal design methods.The adventitious bud regenerating system using leaf sheaths of C.×generalis was successfully established in this study.The results showed that the optimal medium for callus induction was MS+6.62 mg·L^(-1)6-benzylaminopurine(6-BA)+1.5 mg·L^(-1)naphthalene acetic acid(NAA)+0.2 mg·L^(-1)thidiazuron(TDZ),with an induction rate of 80.77%.The best condition for callus proliferation was dark culture for 14 days.The best medium for C.×generalis callus proliferation was MS+2.3 mg·L^(-1)6-BA+2.28 mg·L^(-1)TDZ+0.1 mg·L^(-1)3-indoleacetic acid(IAA),the proliferation rate was 97.50%,and the proliferation coefficient was 4.28.The optimal medium for adventitious bud differentiation was MS+1.0 mg·L^(-1)6-BA+0.1 mg·L^(-1)IAA,with a differentiation rate of 88.00%.The optimal medium for adventitious shoots multiplication was MS+6 mg·L^(-1)6-BA+1.5 mg·L^(-1)NAA+0.2 mg·L^(-1)TDZ,and the proliferation coefficient could be 18.00.In addition,sterile seedlings were grown in 1/2MS+0.5 mg·L^(-1)NAA+30 g·L^(-1)sucrose medium for rooting culture,and the rooting rate was 100%.Moreover,the survival rate of seedlings transplanted in a 1:1:1 volume ratio of grass charcoal–organic fertilizer–soil mixture was 98.33%.Over all,the regenerating system of C.×generalis established in this study could provide technical support for germplasm resource conservation,genetic transformation system establishment,genetic improvement,and new variety breeding of C.×generalis.
作者
甘祖燕
潘学军
李琴
舒木林
段玫
张文娥
GAN Zuyan;PAN Xuejun;LI Qin;SHU Mulin;DUAN Mei;ZHANG Wen'e(College of Agriculture,Guizhou University,Guiyang 550025,China)
出处
《植物生理学报》
CAS
CSCD
北大核心
2023年第8期1543-1554,共12页
Plant Physiology Journal
基金
贵州省科技成果转化项目(黔科合成果[2021]一般009)
贵州省高层次创新型人才培养项目(黔科合人才[2016]4038号)
国家自然科学基金(31460371)。
关键词
不定芽
大花美人蕉
愈伤诱导
分化
再生体系
adventitious bud
Canna×generalis
callus induction
differentiation
regeneration system
