蔷薇科果树CLE多肽家族的鉴定及梨PbrCLE31调控花粉管生长功能分析  

Identification of CLE peptide family in Rosaceae fruit trees and regulation of pollen tube growth by PbrCLE31 in pear

作　　者：程梦雨 李小强 王鹏[1] 张华[2] 张绍铃 吴巨友 CHENG Mengyu;LI Xiaoqiang;WANG Peng;ZHANG Hua;ZHANG Shaoling;WU Juyou(College of Horticulture/State Key Laboratory of Crop Genetics and Germplasm Innovation,Nanjing Agricultural University,Nanjing 210095,China;Shanghai Vocational College of Agriculture and Forestry,Shanghai 201699,China;Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement,Nanjing 210014,China)

机构地区：[1]南京农业大学园艺学院/作物遗传与种质创新国家重点实验室,江苏南京210095 [2]上海农林职业技术学院,上海201699 [3]江苏省高效园艺作物遗传改良重点实验室,江苏南京210014

出　　处：《南京农业大学学报》2021年第5期850-861,共12页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金项目(31772256);上海市教育发展基金会“晨光计划”项目(11CGB18)。

摘　　要：[目的]本文旨在鉴定分析CLE(CLAVATA3/Embryo surrounding region)多肽家族的特点及进化特征,深入了解蔷薇科果树CLE多肽的功能。[方法]基于全基因组测序结果,采用生物信息学方法,参考拟南芥中CLE多肽序列鉴定梨、苹果、桃、梅、草莓和黑树莓6个蔷薇科果树物种CLE多肽家族序列信息,重点分析梨CLE多肽家族的基因结构和理化特性等;通过转录组数据分析PbrCLE成员在不同组织以及在花柱和花粉不同发育阶段的表达模式;并通过实时荧光定量PCR(RT-qPCR)技术分析5个PbrCLE成员在花粉和花柱中的表达模式。通过外源CLE多肽处理梨花粉,研究其调控花粉管生长的功能。[结果]本研究在蔷薇科果树中共筛选到91个CLE候选基因。各物种CLE基因不均匀分布在5~12条染色体上。聚类分析将所有CLE基因分为4组。K a/K s分析表明大部分PbrCLE基因都经历了纯化选择。转录组结果表明大部分CLE基因在梨不同组织中的表达量均较低,这与多肽信号分子在植物体内含量较低相一致。外源合成的PbrCLE31多肽可以抑制体外培养的梨花粉管生长。[结论]蔷薇科果树中存在保守的CLE基因,并且PbrCLE31多肽可以抑制梨花粉管的生长。[Objectives]This article aimed to identify and analyze the characteristics and evolutionary characteristics of the CLE(CLAVATA3/Embryo surrounding region)peptide family,and to gain an in-depth understanding of the functions of CLE peptides from Rosaceae fruit trees.[Methods]Based on the results of whole-genome sequencing,using bioinformatics methods and referring to the CLE peptide sequences in Arabidopsis thaliana,the CLE peptide family sequences information was identified in fruit trees of Rosaceaes,including Pyrus bretschneideri,Malus domestica,Prunus persica,Prunus mume,Fragaria vesca and Rubus occidentalis.The gene structure and physical and chemical properties of the pear CLE peptide family were analyzed,the expression patterns of PbrCLE members in different tissues and at different developmental stages of style and pollen through transcriptome data were analyzed,and the expression patterns of 5 PbrCLE member in pollen or style were analyzed by real-time fluorescent quantitative PCR(RT-qPCR)technology.The pear pollen was processed by exogenous CLE peptide to study its function of regulating the growth of pollen tube.[Results]91 CLE candidate genes were screened in Rosaceae fruit trees.The CLE genes of various species were unevenly distributed on 5-12 chromosomes.Cluster analysis divided all CLE genes into 4 groups.K a/K s analysis showed that most of the PbrCLE genes had undergone purification selection.The transcriptome results showed that the expression levels of most CLE genes in different tissues of pears were low,which was consistent with the low content of peptide signal molecules in plants.Exogenously synthesized PbrCLE31 peptide could inhibit the growth of pear pollen tubes cultured in vitro.[Conclusions]There was conservative CLE gene in Rosaceae fruit trees,and the PbrCLE31 peptide could inhibit the growth of pear pollen tubes.

关 键 词：梨 CLE多肽 蔷薇科果树 CLE基序 表达分析 

分 类 号：S661.2[农业科学—果树学]