烟草胚珠单倍体植株加倍与快繁一体化技术研究  

Integrated technology of doubling and rapid propagation of tobacco ovule haploid plants

作　　者：曹景林 王欣 程君奇 李亚培 吴成林 张俊杰 蔡长春 CAO Jinglin;WANG Xin;CHENG Junqi;LI Yapei;WU Chenglin;ZHANG Junjie;CAI Changchun(Tobacco Research Institute of Hubei Province,Wuhan 430030,China)

机构地区：[1]湖北省烟草科学研究院,武汉430030

出　　处：《烟草科技》2024年第4期32-41,共10页Tobacco Science & Technology

基　　金：中国烟草总公司湖北省公司科技重点项目“烤烟抗青枯病、赤星病新品系生物育种与示范”(027Y2022-021)。

摘　　要：为探讨胚珠单倍体植株的染色体加倍及其双单倍体快繁技术,以烟草雄性不育杂交种KRK26的胚珠单倍体植株为材料,选取6种发育阶段的叶片,以其主脉切段作为外植体,采用细胞培养加倍法对胚珠单倍体植株进行染色体加倍。结果表明,采用体细胞培养加倍法对胚珠单倍体植株进行染色体加倍优于秋水仙碱浸苗法;在不同发育阶段的叶片中,以完全展开达21~28 d的叶片的主脉切段所诱导再生植株的染色体加倍频率最高,达50.15%~54.06%;同一发育阶段的叶片,其主脉切段所诱导再生植株的染色体加倍频率高于叶肉组织小块所诱导再生植株的染色体加倍频率;同一发育阶段的叶片,其主脉基部至近主脉基部1/4处的主脉切段所诱导再生植株的染色体加倍频率最高,达81.79%;通过流式细胞仪鉴定筛选出的双单倍体幼苗经水培炼苗14 d左右,烟苗移栽成活率可达100%。建立了胚珠单倍体植株染色体加倍与快繁一体化育苗技术体系。To investigate the chromosome doubling and double haploid rapid propagation technique of ovule haploid plants,six developmental stages of leaves from ovule haploid plants of male sterile tobacco hybrid KRK26 were selected,and their main vein segments were used as explants to double the chromosomes of ovule haploid plants by the cell culture doubling method.The results showed that chromosome doubling of ovule haploid plants by somatic cell culture was superior to the colchicine soaking method.Among leaves at different developmental stages,the chromosome doubling frequency of regenerated plants induced by the main vein segment of leaves,which remained on the plant for 21 to 28 days after full unfolding,was the highest,reaching 50.15%-54.06%.At the same developmental stage,the chromosome doubling frequency of regenerated plants induced by the main vein segments of leaves was higher than that induced by the mesophyll tissue fragments of leaves;that induced by the main vein segment from the base of the main vein to about 1/4 of the base of the main vein in leaves was the highest,reaching 81.79%.The transplantation survival rate of the double haploid seedlings selected by flow cytometry identification could reach 100%after hydroponic cultivation for about 14 days.An integrated technology system for chromosome doubling and rapid propagation of ovule haploid plants was established.

关 键 词：烟草 胚珠单倍体 染色体加倍 双单倍体 快繁技术 

分 类 号：S572.01[农业科学—烟草工业]