摘要
自交授粉后花粉管程序化死亡是植物自交不亲和的重要机制,乙烯在细胞程序化死亡过程中具有重要的调控作用。本研究以油茶(Camellia oleifera)良种‘华硕’为材料,通过逆转录PCR (RT-PCR)方法克隆出油茶乙烯受体蛋白(ETR)基因家族的3个成员,分别命名CoETR1、CoETR2.1和CoETR2.2,基因编码区长度分别为2 220、2 292和2 292 bp,分别编码740、764和764个氨基酸,含有3~4个跨膜结构域。氨基酸比对和进化树分析结果表明, CoETR1与CoETR2.1、CoETR2.2同源性分别为53.4%、53.1%;三个蛋白与茶树(C. sinensis) ETR1、ETR2.1和ETR2.2的同源性分别为98.0%、98.8%和97.6%,与进化树分析结果一致。利用MEME在线工具对建树的3个蛋白进行保守基序(motif)的预测和分析,结果显示其中10个基序是ETR家族的保守基序, motif 13是ETR1的特征基序, motif 14是ETR2的特征基序。亚细胞定位分析表明三个基因均定位在内质网上。实时荧光定量PCR (qRT-PCR)结果显示自交授粉后36 h时, CoETR1、CoETR2.1和CoETR2.2在子房中表达量达到最高,且显著高于其他组合和时间段。本研究为进一步研究乙烯受体在油茶自交花粉管程序化死亡过程中的作用提供参考。
Programmed cell death(PCD) of pollen tubes after self-pollination is an important mechanism of plant self-incompatibility, and ethylene plays an important regulatory role in PCD. Using Camellia oleifera ‘Huashuo’ as material, three members of ethylene receptor protein gene family were cloned by RT-PCR and named CoETR1, CoETR2.1 and CoETR2.2, respectively. The results show that the coding regions of CoETR1, CoETR2.1 and CoETR2.2 were 2 220, 2 292 and 2 292 bp, encoding 740, 764 and 764 amino acids, and containing 3 or 4 transmembrane domains, respectively. Alignment of amino acid sequences indicates that CoETR1 share 53.4% and 53.1% homology with CoETR2.1 and CoETR2.2, respectively, but had the highly homology with ETR1(98.0%), ETR2.1(98.8%), and ETR2.2(97.6%) in tea(C. sinensis), which were consistent with the results of phylogenetic tree analysis. MEME results showed that 10 motifs were conserved motifs of ETR family, motif 13 was characteristic motif of ETR1 and motif 14 was characteristic motif of ETR2. All the CoETR genes were located on the endoplasmic reticulum by analyzing for subcellular localization. The results of quantitative real-time PCR showed that the expression of CoETR1, CoETR2.1 and CoETR2.2 reached highest levels at 36 h in the ovary after self-pollination, and were significantly higher than other treatments and time. This study provides a reference for the future study of ethylene receptors’ role on PCD of self-pollinated pollen tube in C. oleifera.
作者
周俊琴
卢梦琪
余姝姝
刘懿瑶
杨进
谭晓风
ZHOU Junqin;LU Mengqi;YU Shushu;LIU Yiyao;YANG Jin;TAN Xiaofeng(Central South University of Forestry and Technology,Key Laboratory of Cultivation and Protection for Non-wood Forest Trees,Ministry of Education,Changsha 410004,China)
出处
《植物生理学报》
CAS
CSCD
北大核心
2020年第4期721-733,共13页
Plant Physiology Journal
基金
国家自然科学基金重点项目(31730016)
湖南省研究生科研创新项目(CX2018B440)
中南林业科技大学研究生科技创新基金资助项目(20181014)。
关键词
油茶
自交不亲和
乙烯受体
亚细胞定位
基因表达
oiltea
self-incompatibility
ethylene receptor
subcellular localization
gene expression
