剑麻PGIP基因克隆和表达研究  

Cloning and Expression Analysis of Polygalacturonase-inhibiting Protein in Sisal

作　　者：张燕梅[1] 王瑞芳 杨子平[1] 李俊峰[1] 鹿志伟 赵艳龙[1] 陆军迎[1] 周文钊[1] ZHANG Yanmei;WANG Ruifang;YANG Ziping;LI Junfeng;LU Zhiwei;ZHAO Yanlong;LU Junying;ZHOU Wenzhao(South Subtropical Crops Research Institute,Chinese Academy of Tropical Agricultural Sciences/Zhanjiang City Key Laboratory for Tropical Crops Genetic Improvement,Zhanjiang,Guangdong 524091,China)

机构地区：[1]中国热带农业科学院南亚热带作物研究所/湛江市热带作物遗传改良重点实验室

出　　处：《热带作物学报》2019年第12期2397-2404,共8页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金项目(No.31401427);国家麻类产业技术体系项目(No.CARS-16);广东省甘蔗剑麻产业技术体系创新团队(2019KJ104-03)

摘　　要：多聚半乳糖醛酸酶抑制蛋白(polygalacturonase inhibitor proteins, PGIPs)是一类与植物自身免疫相关的多功能蛋白,在植物防卫反应中扮演着重要角色。为了探讨剑麻PGIP基因的功能,本研究利用PCR的方法从剑麻H.11648中克隆2个剑麻PGIP基因--AhPGIP1和AhPGIP2。利用荧光定量PCR(qRT-PCR)分析AhPGIP1和AhPGIP2基因在烟草疫霉侵染、伤害、低温、盐胁迫、水杨酸(SA)和茉莉酸甲酯(MeJA)处理后的表达模式。结果表明:AhPGIP1基因cDNA全长为1008 bp,编码335个氨基酸,蛋白分子量约为36.7 kDa,等电点为8.65。AhPGIP2基因全长为981 bp,编码326个氨基酸,蛋白分子量约为35.8kDa,等电点为8.98。AhPGIP1基因在烟草疫霉侵染过程中表达水平先下降后明显上升,侵染48h达到最大值,在盐、伤、SA和MeJA处理后表达水平明显上升,分别在3、12、3、12h达到最大值,低温处理6 h表达水平不变,3、12、24 h表达水平明显下降。AhPGIP2基因在烟草疫霉侵染24、36、48 h表达水平明显下降,侵染72 h明显上升并达到最大值,在盐胁迫、低温、SA和MeJA处理后表达水平明显上升,分别在3、24、3、12h达到最大值,在伤处理12h后显著上升并达到最高水平,在3、6、24h明显下降。本研究为深入探讨剑麻PGIP基因在不同逆境胁迫反应中的功能奠定了基础。Polygalacturonase-inhibiting proteins(PGIPs), a group of plant defense proteins, play a crucial role in plant defense reaction. In order to excavate the important function of AhPGIPs, the study cloned AhPGIP1 and AhPGIP2 from H.11648 by PCR. The expression levels of AhPGIP1 and AhPGIP2 were analyzed in sisal H.11648 under several treatments, including Phytophthora nicotianae Breda, methyl jasmonate(MeJA), salicylic acid(SA), salt, low temperature and wounding by fluorescence quantitative PCR(qRT-PCR) technique. The results showed that the full-length cDNA of AhPGIP1 contained 1008 bp and was predicted to encode a protein of 335 amino acids with a theoretical molecular mass of 36.7 kDa and pI of 8.65. The full-length cDNA of AhPGIP2 contained 981 bp, and was predicted to encode a protein of 326 amino acids with a theoretical molecular mass of 35.8 kDa and pI of 8.98. The expression level of AhPGIP1 was not changed at 24 h and then significantly up-regulated by Phytophthora nicotianae Breda, and reached the max value after inoculating 48 h, and was up-regulated by MeJA, SA, salt, and wounding, and reached the max value after stresses 3, 12, 3 and 12 h, respectively. The expression level of AhPGIP1 was not changed by low temperature at 6 h and significantly down-regulated at 3, 12 and 24 h. The expression level of AhPGIP2 was significantly down-regulated at 24, 36 and 48 h and up-regulated at 72 h by Phytophthora nicotianae Breda, was significantly up-regulated by salt, low temperature, SA and MeJA, and reached the max value after stresses 3, 24, 3 and 12 h respectively. The expression level of AhPGIP2 was significantly up-regulated and reached the max value after stress 12 h by wounding, and significantly down-regulated after stress 3, 6, and 24 h. The study laid a solid foundation for further exploration of the function of AhPGIPs.

关 键 词：剑麻 QRT-PCR 基因表达 PGIP 

分 类 号：S563.8[农业科学—作物学]