山茶叶片的离体再生及内源激素与不定芽分化的关系  

Establishment of Regeneration in Vitro System of Camellia japonica and Relationship Between Endogenous Hormones and Adventitious Bud Differentiation

作　　者：王建昭 高园 刀梅 张洪烨 杨自云[1] 陈龙清 吴田[1] WANG Jianzhao;GAO Yuan;DAO Mei;ZHANG Hongye;YANG Ziyun;CHEN Longqing;WU Tian(Yunnan Province Engineering Research Center for Functional Flower Resources and Industralization,School of Landscape Architecture and Horticulture Sciences,Southwest Forestry University,Kunming 650224,China)

机构地区：[1]西南林业大学园林园艺学院,云南省功能性花卉资源及产业化技术工程研究中心,昆明650224

出　　处：《园艺学报》2024年第8期1891-1905,共15页Acta Horticulturae Sinica

基　　金：云南省农业基础研究联合专项面上项目(202301BD070001-242);国家重点研发专项(2019YFD1001005);云南省教育厅研究生项目(2024Y587)。

摘　　要：以山茶(Camellia japonica)叶片为外植体进行愈伤组织诱导和再分化。结果表明:叶片在MS+5.0 mg·L^(-1)6-BA+0.1 mg·L^(-1)2,4-D+5.0 mg·L^(-1)PVP+3%蔗糖培养基上愈伤组织诱导率最高(79.19%);将愈伤组织转至原配方培养基光照培养(12 h·d^(-1)),可分化出不定芽,分化率为51.32%。不定芽在不定根诱导培养基(1/8MS+0.5 mg·L^(-1)IBA+1.0 mg·L^(-1)PVP+2%蔗糖)上生根率最高(50%);生根试管苗移栽至田间,成活率达66.67%。利用液相色谱串联质谱法(LC–MS/MS)检测激素发现,具不定芽分化能力的愈伤组织中,激素信号转导途径、玉米素生物合成代谢途径、α–亚麻酸代谢途径激素含量发生显著变化。在所有检测的内源激素中,有30种(90.9%)含量下调,仅有3种激素含量上调。结果显示,在培养条件一致的前提下,内源激素含量是决定山茶不定芽分化的主要因素。A regeneration system has been established in this study using Camellia japonica leaves as explants.The highest frequency of callus induction(79.19%)was obtained by culturing the leaf explants on the MS media containing 5.0 mg·L^(-1)6-BA,0.1 mg·L^(-1)2,4-D,5.0 mg·L^(-1) PVP and 3%sucrose.Differentiated adventitious buds were generated by directly transferring the explants to the condition of light(12 h·d^(-1)).The frequency of adventitious buds generation was 51.32%.Consequently,adventitious roots were induced from these buds using 1/8MS media containing 0.5 mg·L^(-1) IBA,1.0 mg·L^(-1) PVP and 2%sucrose.The highest frequency of root induction was 50%.The C.japonica seedlings rate was 66.67%.The endogenous hormone levels in the tissue-cultured materials were detected using LC–MS/MS.The results demonstrated that the plant hormone signal transduction pathway,zeatin biosynthesis pathway andα-linolenic acid metabolic pathway were changed in the callus with ability to differentiate adventitious buds.Among all the endogenous hormones detected,a total of 30 were down-regulated(90.9%),only three hormones showed up-regulation.The endogenous hormone levels largely determined the induction of callus and differentiation of adventitious buds of C.japonica explants grown under the same culture conditions.

关 键 词：山茶 再生体系 内源激素 不定芽分化 

分 类 号：S685.14[农业科学—观赏园艺]