甘蔗亲免素基因ScFKBP12-1的克隆及表达特性分析  

Cloning and Expression Analysis of an Immunophilin Gene ScFKBP12-1 from Sugarcane(Saccharum spp. complex)

作　　者：于庆[1] 汤翰臣 钟小强 马晶晶[1] 许莉萍[1] 郭晋隆[1] Yu Qing;Tang Hanchen;Zhong Xiaoqiang;Ma Jingjing;Xu Liping;Guo Jinlong(Key Laboratory of Sugarcane Biology and Genetic Breeding,Fujian Agriculture and Forestry University,Fuzhou,350002)

机构地区：[1]福建农林大学,农业部福建甘蔗生物学与遗传育种重点实验室,福州350002

出　　处：《分子植物育种》2020年第16期5280-5289,共10页Molecular Plant Breeding

基　　金：国家重点研发计划项目(2018YFD1000503);国家现代农业产业技术体系建设专项(CARS-20)共同资助。

摘　　要：FKBP (FK506 binding proteins)是一类能与免疫抑制剂他克莫司(tacrolimus, FK506)和雷帕霉素(rapamycin, RAPA)特异结合的受体蛋白,以基因家族的形式存在于植物中并参与多种生物进程,是植物亲免素蛋白的成员。本研究通过甘蔗(Saccharum spp. complex)茎cDNA文库测序获得一条亲免素全长基因ScFKBP12-1 (登录号:MN242814)。ScFKBP12-1 cDNA序列全长696 bp,开放读码框长339 bp,共编码112个氨基酸。生物信息学分析显示,ScFKBP12-1推导蛋白含有一个FKBP类型的肽脯氨酰顺反异构酶催化结构域,是定位于细胞质的稳定亲水性蛋白,分子量为12.20 kD。组织特异性表达分析表明,ScFKBP12-1基因在甘蔗根中的相对表达量显著高于其在叶、蔗茎髓部、蔗茎表皮和腋芽中的表达水平。定量PCR结果显示,甘蔗小苗中ScFKBP12-1对RAPA (5μmol/L)的处理不敏感,提示了ScFKBP12失去与RAPA绑定的能力。定量PCR结果进一步显示,ScFKBP12-1的表达水平受PEG (25%)和MeJA (100μmol/L)的诱导显著上调,在处理前期就分别上调为对照的2.75和3.11倍,并在处理周期内分别维持在高于对照1.87和2.00倍的水平。此外,该基因的表达还受到低温4℃,Na Cl (250 mmol/L)和SA (5 mmol/L)处理的诱导,分别在处理24 h、48 h和3 h时上调表达。以上结果提示了ScFKBP12-1参与了甘蔗对外源激素(MeJA和SA)、低温、高盐和PEG模拟干旱胁迫等逆境的应答过程,其功能可能与甘蔗响应渗透胁迫密切相关,并受MeJA信号的正向调控。FKBP(FK506 binding proteins),a member of the immunophilin family,are proteins specific binding to the immunosuppressive agents of tacrolimus(FK506)and rapamycin(RAPA)which comprise a large gene family and are involved in various biological processes.A c DNA sequence of FKBP,termed as ScFKBP12-1(Accession number:MN242814),was cloned from sugarcane(Saccharum spp.complex)stem full-length cDNA library.The full length of the ScFKBP12-1 was 696 bp,containing a 339 bp open reading frame(ORF)and encoding 112 amino acids.Bioinformatics analysis revealed that the deduced protein ScFKBP12-1 was a stable hydrophilic protein with a molecular weight of 12.20 kD.It contained a single PPIase-FKBP type domain and was predicted to localize in the cytoplasm.The real-time qPCR results demonstrated that the expression level of ScFKBP12-1 in sugarcane root was significantly higher than that in the leaf,pith of the stem,and bud.The results also indicated that the seedlings of sugarcane were insensitive to RAPA(5μmol/L),implying ScFKBP12 lost the function to bridge the interaction of RAPA and TOR in sugarcane.The expression level of ScFKBP12-1 significantly up-regulated under the treatments of PEG(25%)and MeJA(100μmol/L),which was 2.75 and 3.11 times higher than control at 12 h and 3 h,respectively,and kept 1.87 and 2.00 times high level,respectively,during the whole treatment stage.Additionally,the expression level of ScFKBP12-1 induced by NaCl(250 mmol/L),chill(4℃),and SA(5 mmol/L)at 24 h,48 h,and 3 h,respectively.These results were suggesting that ScFKBP12-1 was involved in sugarcane responding to exogenous hormones(Me JA and SA),low temperature,salt,and PEG simulated drought stresses,and might play an active role in MeJA-regulated osmotic stress responses.

关 键 词：甘蔗(Saccharum spp.complex) 亲免素 FKBP 非生物胁迫 实时荧光定量PCR 

分 类 号：S566.1[农业科学—作物学]