油茶自、异交雌蕊CAT活性及相关基因克隆表达分析  

CAT activity and related gene expression in the pistil of self-and cross-pollinated Camellia oleifera Abel.

作　　者：蒋思思 周俊琴[1] 卢梦琪 袁军[1] Jiang Si-Si;Zhou Jun-Qin;Lu Meng-Qi;Yuan Jun(Key Laboratory of Cultivation and Protection for Non-wood Forest Trees,Ministry of Education,Key Lab of Non-wood Forest Products of State Forestry Administration,Central South University of Forestry and Technology,Changsha 410004,China)

机构地区：[1]中南林业科技大学,经济林培育与保护教育部重点实验室,经济林育种与栽培国家林业局重点实验室,长沙410004

出　　处：《植物科学学报》2022年第6期810-819,共10页Plant Science Journal

基　　金：湖南省重点研发计划(2020NK2050);国家自然科学基金(31730016);湖南省教育厅科学研究项目(21B0274);中南林业科技大学引进人才科研启动基金(2021YJ018);湖南省研究生科研创新项目(CX20220709);中南林业科技大学研究生科技创新基金(CX202202006)。

摘　　要：油茶(Camellia oleifera Abel.)具有后期自交不亲和(LSI)特性,严重阻碍了油茶杂交育种的亲本选择和品种配置。为研究过氧化氢酶(Catalase,CAT)在油茶自交不亲和中的作用,本文对油茶自、异交授粉后雌蕊中的CAT活性进行了测定,并克隆得到3个CAT基因,命名为CoCAT1~CoCAT3。结果显示,其CDS分别为1479、1287和1479 bp,编码492、428和492个氨基酸。多序列比对结果表明,其编码产物在C末端均含有典型的三肽序列。系统进化分析发现,CoCAT2、CoCAT3与NtCAT3亲缘关系最近,CoCAT1与CsCAT1亲缘关系最近。基因表达模式分析结果显示,3个基因在授粉72 h后的自交雌蕊中表达量最大,在授粉36、84 h自交雌蕊中的表达量显著高于异交。自、异花授粉均引起油茶雌蕊内CAT活性的上升,但幅度不同,推测油茶CAT可能参与油茶自交不亲和的响应机制。Camellia oleifera Abel. shows late-acting self-incompatibility(LSI) characteristics, which seriously hinder parental selection and cultivar arrangement during cross breeding. To study the role of catalase(CAT) in the self-incompatibility of C. oleifera, CAT activities in the pistils after self-and cross-pollination were determined. Three CAT genes(CoCAT1, CoCAT2, and CoCAT3) were cloned by reverse-transcription polymerase chain reaction(RT-PCR), with coding sequences(CDSs)of 1479, 1287, and 1479 bp, encoding 492, 428, and 492 amino acids, respectively. Multiple sequence alignment showed that their coding products contained typical tripeptide sequences at the C-terminal.Phylogenetic analysis showed that CoCAT2 and CoCAT3 were most closely related to NtCAT3, and CoCAT1 was most closely related to CsCAT1. Expression pattern analysis indicated that the expression levels of the three genes were highest in the self-pollinated pistils 72 h after pollination, and expression levels in the self-pollinated pistils at 36 and 84 h were significantly higher than those in the cross-pollinated pistils.Thus, self-pollination and cross-pollination increased CAT activity in the pistils of C. oleifera, but at different magnitudes. This study indicated that CAT maybe involved in the response mechanism of self-incompatibility in C. oleifera.

关 键 词：油茶 自交不亲和 过氧化氢酶 基因克隆 表达模式 

分 类 号：Q943.2[生物学—植物学]