梨FWL5基因克隆及表达分析  

Cloning and Expression Analysis of FWL5 Gene in Pear

作　　者：张艳[1] 田嘉[1] 张峰 温玥 Zhang Yan;Tian Jia;Zhang Feng;Wen Yue(College of Forestry and Horticulture,Xinjiang Agricultural University,Urumqi,830052;Xinjiang Korla Research Center of Fragrant Pear,Korla,841000)

机构地区：[1]新疆农业大学林学与园艺学院,乌鲁木齐830052 [2]新疆库尔勒市香梨研究中心,库尔勒841000

出　　处：《分子植物育种》2022年第15期4940-4947,共8页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31760561);新疆维吾尔自治区园艺重点学科基金项目(2016-10758-3)共同资助。

摘　　要：Fw2.2是调控果实大小的数量性状基因,在果实生长发育过程中扮演重要角色。为探明梨FWL(fruit weight 2.2-like)家族基因在果实发育过程中的作用,本研究以小果型野生杜梨、中果型库尔勒香梨、大果型库尔勒香梨芽变品种‘早美香’以及大果栽培型鸭梨为试验材料,从梨基因组中筛选分离出梨FWL5基因,对该基因进行克隆和序列分析,并利用荧光定量PCR技术检测梨FWL5基因在四个梨品种的果实细胞分裂期表达水平。结果显示,梨FWL5基因开放读码框长度为729 bp,编码了242个氨基酸,为亲水性不稳定蛋白,存在两个跨膜结构。结构域分析发现梨FWL5蛋白包括一个富含半胱氨酸的结构域,属于PLAC8家族基因。系统进化树分析表明,梨FWL5与樱桃Pav CNR12蛋白序列最相近,樱桃PavCNR12蛋白在细胞分裂期能够负调控甜樱桃果实大小,并有可能参与重金属转运。荧光定量PCR检测结果显示,在花期,梨FWL5除在鸭梨盛花期表达量较高,在其余三个梨品种中表达水平相对一致。在幼果期,除‘早美香’外,梨FWL5基因表达水平为杜梨>香梨>鸭梨。本研究结果可为梨FWL5基因在调控果实大小功能方面的研究提供理论依据。Fw2.2 is a quantitative trait gene that regulates fruit size and plays an important role in fruit growth and development.In order to explore the role of FWL(fruit weight 2.2-like)family genes in the fruit development process of pear,in this study,small-fruited wild Duli pear,medium-fruited Korla fragrant pear,large-fruited Korla fragrant pear bud variety‘Zaomeixiang’,and large-fruited Yali were used as test materials,screened and isolated the pear FWL5 gene from the pear genome,cloned and sequenced the gene,used fluorescence Quantitative PCR analysis was used to analyze the expression level of pear FWL5 gene in the fruit cell division of four pear varieties.The results showed that the open reading frame(ORF)of the pear FWL5 gene was 729 bp in length,encoded a predicted protein of 242 amino acids,and the encoded product was an unstable hydrophilic protein with two transmembrane domains.Pear FWL5 protein had a cysteine-rich domain and belongs to the PLAC8 family.Phylogenetic tree analysis showed that the protein sequence of pear FWL5 and cherry PavCNR12 is the closest,and cherry PavCNR12 protein can negatively regulate the size of sweet cherry fruit during cell division and may participate in heavy metal transport.The results of fluorescence quantitative PCR analysis showed that the expression level of the FWL5 gene of the four pear cultivars at the young fruit period was greater than that at the flowering period.In the flowering period,the expression level of pear FWL5 was relatively consistent in the other three pear cultivars except that the expression level of pear FWL5 was higher in the full bloom period.In the young fruit period,except for‘Zaomeixiang’,the expression levels of the FWL5 gene in pears were Duli pear>Korla fragrant pear>Yali pear.The results of this study can provide a theoretical basis for the research on the function of pear FWL5 gene in regulating fruit size.

关 键 词：梨 FWL5基因 基因克隆 表达分析 

分 类 号：S661.2[农业科学—果树学]