油茶未授粉胚珠愈伤组织诱导及形态学和细胞学特征  

Callus induction and morphological and cytological observation of unpollinated ovules of Camellia oleifera

作　　者：靳皓然 杨善为 袁蕾慧子 潘倩 侯思璐 范晓明[1] 袁德义[1] JIN Haoran;YANG Shanwei;YUAN Leihuizi;PAN Qian;HOU Silu;FAN Xiaoming;YUAN Deyi(Key Laboratory of Non-wood Forest Products of National Forestry and Grassland Administration,Central South University of Forestry and Technology,Changsha 410004,Hunan,China;Forestry Bureau of Huaihua City,Huaihua 418099,Hunan,China)

机构地区：[1]中南林业科技大学经济林育种与栽培国家林业和草原局重点试验室,湖南长沙410004 [2]怀化市林业局,湖南怀化418099

出　　处：《浙江农林大学学报》2023年第4期773-782,共10页Journal of Zhejiang A&F University

基　　金：湖南省科技创新计划项目(2021NK1007);国家重点研发计划项目(2018YFD1000603)。

摘　　要：【目的】筛选适合油茶‘华金’Camellia oleifera‘Huajin’未授粉胚珠诱导愈伤组织的最适培养基,并对愈伤组织进行细胞学观察,为油茶单倍体育种提供依据。【方法】以油茶‘华金’为试材,筛选未授粉胚珠诱导愈伤组织的最适培养基配方,研究基础培养基类型MS培养基、木本植物用培养基(WPM)、MT培养基、1/2MS培养基、B5培养基、生长素萘乙酸(NAA)、吲哚乙酸(IAA)、吲哚丁酸(IBA)、二氯苯氧乙酸(2,4-D)和细胞分裂素苄氨基嘌呤(6-BA)、激动素(KT)、噻苯隆(TDZ)组合,以及植物生长调节剂浓度对愈伤组织诱导的影响;选取不同外部形态的愈伤组织进行石蜡切片,并根据流式细胞仪的倍性鉴定结果,研究愈伤组织来源及发育进程。【结果】MS作为基础培养基,添加1.0mg·L^(-1)NAA+2.0 mg·L^(-1)TDZ+40 g·L^(-1)蔗糖的诱导率最高,为92.97%,NAA和TDZ对诱导率影响显著(P<0.05),其中TDZ的影响程度大于NAA。胚性愈伤组织呈浅绿色颗粒状,细胞学特征为排列紧密,核大,倍性鉴定为六倍体。【结论】油茶未授粉胚珠诱导胚性愈伤组织的最适培养基配方为:MS+40 g·L^(-1)蔗糖+7 g·L^(-1)琼脂+1.0 mg·L^(-1)NAA+2.0mg·L^(-1)TDZ;愈伤组织由外珠被细胞脱分化形成。[Objective]The objective is to screen the optimal medium suitable for callus induction from unpollinated ovules of Camellia oleifera,and explore the origin and developmental process of callus by cytological observation,so as to provide data reference for haploid breeding of C.oleifera.[Method]C.oleifera‘huajin’was used as the test material to screen the optimal medium formula for inducing callus from unpollinated ovule,and to study the basic medium types MS,WPM,MT,1/2MS,and B5,as well as effects of plant growth regulator concentration such as auxins NAA,IAA,IBA,and 2,4-D,and cytokinins 6-BA,KT,and TDZ on callus induction.Calluses with different external morphology were selected for paraffin section,and their origin and development process were studied according to the ploidy identification results of flow cytometry.[Result]MS was used as the basic medium,and the highest induction rate was 92.97%when 1.0 mg·L^(−1)NAA+2.0 mg·L^(−1) TDZ+40 g·L^(−1) sucrose was added.NAA and TDZ had significant effects on the induction rate(P<0.05),and the impact of TDZ was greater than that of NAA.The embryogenic callus was light green granular,with cytological characteristics of close arrangement,large nucleus,and ploidy identification of hexaploid.[Conclusion]The optimal medium for inducing embrygenic callus from unpollinated ovules of C.oleifera is MS+40 g·L^(−1)sucrose+7 g·L^(−1) AGAR+1.0 mg·L^(−1) NAA+2.0 mg·L^(−1) TDZ.The callus is formed by dedifferentiation of outer integument cells.

关 键 词：油茶 未授粉胚珠 愈伤组织 细胞学特征 倍性鉴定 

分 类 号：S718.46[农业科学—林学]