海南地区油茶未成熟胚分生结节的植株再生  

Inducing plant regeneration by Camellia vietnamensis immature embryo meristematic nodules in Hainan

作　　者：侯辛辛 陈健妙 高兴[4] 赖杭桂 王健[5] 胡新文[3] 吴友根[5] HOU Xin-xin;CHEN Jian-miao;GAO Xing;LAI Hang-gui;WANG Jian;HU Xin-wen;WU You-gen(College of Forestry,Hainan University,Haikou 570228,China;Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresources,Haikou 570228,China;College of Tropical Crops,Hainan University,Haikou 570228,China;Administration for Market Regulation of Zhuzhou,Zhuzhou,Hunan 412000,China;College of Horticulture,Hainan University,Haikou 570228,China)

机构地区：[1]海南大学林学院,海口570228 [2]海南省热带生物资源可持续利用重点实验室,海口570228 [3]海南大学热带作物学院,海口570228 [4]株洲市市场管理监督局,湖南株洲412000 [5]海南大学园艺学院,海口570228

出　　处：《南方农业学报》2020年第4期867-878,共12页Journal of Southern Agriculture

基　　金：海南省自然科学基金面上项目(319MS014);海南省重大科技计划项目(ZDKJ2017004);海南大学“三全育人”综合改革专项项目(19SQYR08)。

摘　　要：【目的】建立海南地区油茶未成熟胚再生体系,为油茶组培快繁和遗传转化研究提供技术支持。【方法】以海南本地油茶合子胚为外植体,对其进行初代诱导、分化和增殖培养,筛选出最适于诱导分化的外植体胚发育时期,明确植株再生途径,从而建立分生结节组织植株再生体系。【结果】(1)盛花期后240~300 d的油茶幼胚在改良MS+激动素(KT)2.0 mg/L+萘乙酸(NAA)0.2 mg/L+玉米素(ZT)5.0 mg/L培养基上,初代诱导率在90.0%以上,以分生结节组织为主;(2)分生结节组织的适宜增殖培养基为改良MS+KT 1.0~2.0 mg/L+NAA 0.2 mg/L,增殖系数约4.6;(3)未成熟胚分生结节组织的分化培养基为改良MS+6-苄氨基嘌呤(6-BA)2.0 mg/L+NAA 0.2 mg/L+吲哚-3-乙酸(IAA)0.2 mg/L+ZT 5.0 mg/L+酸水解酪蛋白(CH)400.0 mg/L,平均每块分生结节组织诱导不定芽11.3个,不定芽以无糖暴露培养生根方式生根,移栽成活率为91.6%;(4)分生结节组织在改良MS+6-BA 2.0 mg/L+NAA 0.2 mg/L+ZT 5.0 mg/L+CH 400.0 mg/L上以分化体细胞胚状体为主,每块分生结节分化6.5个体细胞胚状体,具双子叶的体细胞胚状体在改良MS+6-BA 0.2 mg/L+IAA 2.0 mg/L+赤霉酸(GA3)0.5 mg/L+CH 200.0 mg/L培养基上有效成熟和萌发,萌发率为72.1%,胚芽和胚根有效伸长形成体胚再生植株,移栽成活率为94.7%。【结论】以油茶未成熟合子胚为外植体,通过分生结节组织方式增殖,可经体细胞胚状体发生和不定芽发生两种途径成功建立海南油茶未成熟胚分生结节植株再生体系。【Objective】To establish immature zygotic embryos regeneration system of Camellia vietnamensis,which provided technical support for the research on rapid propagation and genetic transformation of C.vietnamensis.【Method】Hainan native C.vietnamensis zygotic embryos were used as explants,zygotic embryos were induced to differentiate and proliferate to select the most suitable stage for inducing differentiation.Meanwhile,and the proliferation and plant regeneration pathways were determined,so as to establish the regeneration system of meristematic nodules induced from immature zygotic embryos of C.vietnamensis.【Result】(1)When the immature embryo at 240-300 d after full flowering stage cultured at the modified MS+kinetin(KT)2.0 mg/L+naphthaleneacetic acid(NAA)0.2 mg/L+zeatin(ZT)5.0 mg/L medium,the initial induction rate of the immature embryo was more than 90.0%and the meristematic nodules were majority.(2)The suitable medium for the proliferation of meristematic nodule tissues was the modified MS+1.0-2.0 mg/L KT+0.2 mg/L NAA,and the proliferation coefficient was approximately 4.6.(3)The differentiation medium for immature embryo meristematic nodules was modified MS+2.0 mg/L 6-benzyl adenine(6-BA)+0.2 mg/L NAA+0.2 mg/L indole-3-acetic acid(IAA)+ZT 5.0 mg/L and the average adventitious bud number was 11.3.Adventitious shoots rooted by sugar-free exposed cultivation,and plant transplant survival rate was 91.6%.(4)The meristematic nodule cultured at the modified MS+2.0 mg/L 6-BA+0.2 mg/L NAA+ZT 5.0 mg/L mainly differentiated into somatic embryo with the average somatic embryo number of 6.5.Somatic embryos with dicotyledonate were effectively germinated and developed into plantlets on modified MS+0.5 mg/L 6-BA+2.0 mg/L IAA+0.5 mg/L gibberellic acid(GA3)+CH 200.0 mg/L medium.The germination rate was 72.1%,the germ and the radicle were effectively extended to form a somatic embryo regeneration plant and plant transplant survival rate was 94.7%.【Conclusion】With the immature zygotic embryos as explants,two rege

关 键 词：油茶 未成熟胚 分生结节组织 不定芽 体细胞胚状体 海南 

分 类 号：S794.4[农业科学—林木遗传育种]