摘要
为探究一个受低温诱导的香蕉MaTIFY9基因的序列特征及在不同激素和逆境胁迫处理下的表达模式,分别使用PCR和反转录PCR(RT-PCR)技术克隆该基因的gDNA和cDNA序列,分析其核苷酸和编码蛋白序列特性,利用实时荧光定量PCR(qRT-PCR)技术研究其在不同器官和不同激素、逆境胁迫处理下的表达情况.结果显示:MaTIFY9编码了一个分子式为C_(841)H_(1346)N_(236)O_(249)S_(7),氨基酸大小为172 aa,分子量为18 989.93,等电点为9.55的不稳定亲水性蛋白.MaTIFY9无信号肽和跨膜结构,具有TIFY和Jas(CCT-2)两个保守结构域,属于JAZ亚家族,与小果野蕉TIFY9亲缘关系最近.亚细胞定位预测结果显示MaTIFY9主要定位于细胞核. MaTIFY9启动子含有大量的光响应相关元件,此外还含有许多防御及应激、SA和ABA响应元件. qRT-PCR结果显示MaTIFY9在根中表达量最高,其次是假茎和球茎,叶最低;其表达受脱落酸ABA、水杨酸SA和枯萎病显著诱导,受高温抑制.本研究表明,与其他植物TIFY类似,MaTIFY表达受多种激素和逆境调控,可能在香蕉响应非生物和生物逆境胁迫过程中发挥重要作用.
We investigated the sequence characteristics and expression patterns of a low temperature inducible banana gene, MaTIFY9, under different phytohormone and abiotic stress treatments. To do this, the gDNA and cDNA sequences of the gene were successfully amplified by PCR and reverse transcription PCR(RTPCR), respectively. Thereafter, a series of bioinformatic analyses were performed to reveal the characteristics of its nucleotide and encoded protein sequences. The gene expression patterns in different tissues and organs were analyzed under different treatments using quantitative real-time PCR(qRT-PCR). MaTIFY9 was found to encode for an unstable hydrophilic protein of 172 aa with a molecular formula of C(841)H_(1346)N_(236)O_(249)S_(7), a molecular weight(Mr) of 18 989.93, and an isoelectric point of 9.55. The protein had no signal peptide or transmembrane structure but had two conserved domains(TIFY and Jas) and is thus suggested to be a member of the JAZ(Jasmonate ZIM-domain) protein subfamily. Phylogenic relationship analysis revealed that the MaTIFY9 protein shared its closest relationship with the TIFY9 protein of Musa acuminata subsp. malaccensis. The protein subcellular localization prediction results showed that MaTIFY9 was mainly located in the nucleus. The promoter of MaTIFY9 was shown to contain many light responsive elements, as well as numerous defense and stress-,salicylic acid(SA)-, and abscisic acid(ABA)-responsive elements. qRT-PCR results showed that MaTIFY9 expression was highest in the roots, followed by the pseudostem and corm, while the lowest expression was found in the leaves. MaTIFY9 expression was significantly induced by ABA, SA, and Fusarium wilt treatments,but was inhibited by high temperatures. Similar to TIFY genes in other plant species, MaTIFY9 was found to be responsive to various phytohormone and stress treatments, and thus is likely to play an important role in banana responses to both abiotic and biotic stresses.
作者
刘嘉鹏
谭秋月
伍俊为
王斌
田娜
刘范
孙雪丽
程春振
LIU Jiapeng;TAN Qiuyue;WU Junwei;WANG Bin;TIAN Na;LIU Fan;SUN Xueli;CHENG Chunzhen(College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou 350002,China)
出处
《应用与环境生物学报》
CAS
CSCD
北大核心
2021年第6期1609-1615,共7页
Chinese Journal of Applied and Environmental Biology
基金
国家重点研发计划课题(2019YFD1000901)
国家自然科学基金项目(31601713)
国家现代农业产业技术体系建设专项资金项目(CARS-32-11)资助。
