‘库尔勒香梨’PsiCAD1和PsiPRX6基因的亚细胞定位和表达分析  

Subcellular Localization and Expression Analysis of PsiCAD1 and PsiPRX6 Genes in'Kuerle Pear'

作　　者：张倩 贡鑫 张天正 陈燕 王浩 吴翠云[1,2] 陶书田 包建平[1,2] Zhang Qian;Gong Xin;Zhang Tianzheng;Chen Yan;Wang Hao;Wu Cuiyun;Tao Shutian;Bao Jianping(College of Plant Science,Tarim University,Alar,843300;National and Local Joint Engineering Laboratory of High-efficiency and Superior-quality Cultivation and Fruit Deep Processing Technology of Characteristic Fruit trees in South Xinjiang,Key Laboratory of Protection and Utilization of Bio-logical Resources in Tarim Basin,Xinjiang Production and Construction Corps,Tarim University,Alar,843300;College of Horticulture,Nanjing Agricultural University,Nanjing,210095)

机构地区：[1]塔里木大学植物科学学院,阿拉尔843300 [2]塔里木大学,新疆生产建设兵团塔里木盆地生物资源保护利用重点实验室,南疆特色果树高效优质栽培与深加工技术国家地方联合工程实验室,阿拉尔843300 [3]南京农业大学园艺学院,南京210095

出　　处：《分子植物育种》2023年第18期5947-5954,共8页Molecular Plant Breeding

基　　金：新疆生产建设兵团财政专项-南疆科研条件建设计划及重点产业创新发展支撑计划-科技发展专项(2017-DB006-3);新疆生产建设兵团塔里木盆地生物资源保护利用重点实验开放课题(BRYB1903);南京农业大学-塔里木大学科研联合基金项目(NNTDLH201904,NNLH202007)共同资助。

摘　　要：石细胞含量的多少是影响梨果实口感关键因素之一,而木质素是梨石细胞的主要组成成分,肉桂醇脱氢酶(CAD)和过氧化物酶(PRX)是合成木质素关键的酶。为了探索CAD和PRX基因在‘库尔勒香梨’石细胞发育过程中的作用,本研究以‘库尔勒香梨’为试验材料,测定梨果实石细胞含量与木质素含量,并运用亚细胞定位与瞬时表达技术进行PsiCAD1和PsiPRX6的功能验证。结果表明‘库尔勒香梨’果实石细胞含量与木质素含量在30~80 DAF呈先上升后下降的趋势,同时通过RT-qPCR发现PsiCAD1、PsiPRX2、PsiPRX6与PsiPRX17不同时期的表达量与果实石细胞含量和木质素含量发育趋势一致;通过亚细胞定位技术发现Psi CAD1-GFP与PsiPRX6-GFP融合蛋白荧光信号在细胞膜中表达,推测该蛋白激酶合成后可能在质膜上发挥作用;在梨果中瞬时表达过表达载体中发现PsiPRX6、PsiCAD1过表达下观察到木质素染色与含量均增加,且注射位点的PsiCAD1与PsiPRX6的表达量也有所增加。上述结果初步表明PsiCAD1与PsiPRX6基因参与调控木质素的合成,为进一步了解木质素合成和细胞壁木质化之间的关系提供了重要理论依据。The amount of stone cells is one of the key factors affecting the real taste of pear fruit,lignin is the main component of pear cells,cinnamol dehydrogenase(CAD)and peroxidase(PRX)are key enzymes for the synthesis of lignin.To explore the role of CAD and PRX genes in the development of'Kuller Pear'stone cells,using'Kuller Pear'as test material,determination of cell content and lignin content of pear fruit solid stone.Subcellular localization and transient expression were used to verify the function of PsiCAD1 and PsiPRX6.The results showed that the content of fruit stone cells and lignin decreased at 30~80 DAF.Meanwhile,it was found by RT-qPCR that the expression of PsiCAD1,PsiPRX2,PsiPRX6 and PsiPRX17 in different periods was consistent with the development trend of fruit stone cell content and lignin content;using subcellular localization techniques,PsiCAD1-GFP and PsiPRX6-GFP fusion proteins were expressed in cell membranes.The above results initially show that the PsiCAD1 and PsiPRX6 gene are involved in the regulation of lignin synthesis,providing an important theoretical basis for further understanding the relationship between lignin synthesis and lignization of the cell wall.

关 键 词：梨 PsiCAD1 PsiPRX6 亚细胞定位 瞬时表达 

分 类 号：S661.2[农业科学—果树学]