|本期目录/Table of Contents|

 QUE Wancai,HUANG Ning,LIU Feng,et al.Isolation and expression of a eukaryotic translation initiation factor 5A gene from sugarcane[J].Chinese Journal of Applied & Environmental Biology,2015,21(06):1120-1127.[doi:10.3724/SP.J.1145.2015.04008]





Isolation and expression of a eukaryotic translation initiation factor 5A gene from sugarcane
阙万才 黄宁 刘峰 肖新换 凌辉 张玉叶 苏炜华 苏亚春 吴期滨 阙友雄
1福建医科大学附属协和医院药学部 福州 350001 2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/国家甘蔗产业技术研发中心 福州 350002
QUE Wancai HUANG Ning LIU Feng XIAO Xinhuan LING Hui ZHANG Yuye SU Weihua SU Yachun WU Qibin QUE Youxiong
1Department of Pharmacy, Fujian Medical University Union Hospital, Fuzhou 350001, China 2Key Laboratory of Sugarcane Biology and Genetic Breeding (Fujian), Ministry of Agriculture, Fujian Agriculture and Forestry University/Sugarcane Research & Development Center, China Agricultural Technology System, Fuzhou 350002, China
sugarcane eIF5A Sporisorium scitamineum bioinformatics Real-time Quantitative PCR
Q78 : S566.103.4
真核生物翻译起始因子5A(Eukaryotic translation initiation factor 5A,eIF5A)是一种在动植物和真菌体内普遍存在的蛋白质. 为了解eIF5A基因在甘蔗应答生物逆境胁迫中的作用,本研究首先从黑穗病菌胁迫下甘蔗抑制消减杂交(Suppression subtractive hybridization,SSH)文库中获得一条与玉米eIF5A基因(GenBank Accession Number:EU958725.1)同源性为93%的甘蔗EST序列;其次,以此序列作为探针,通过电子克隆技术获得一条甘蔗eIF5A基因的cDNA拼接序列;最后,经RT-PCR扩增和测序验证,结果显示电子克隆序列与测序序列一致,将该序列命名为SceIF5A(GenBank Accession Number:KJ577595). 生物信息学分析显示,SceIF5A基因cDNA全长1 174 bp,含有长度为483 bp、编码160个氨基酸的完整开放读码框;SceIF5A蛋白是酸性稳定蛋白,分子量(Mr)为17 453.6,含有12个保守氨基酸序列,推测定位于细胞质. 实时荧光定量PCR(Real-time quantitative PCR,RT-qPCR)分析结果表明,在黑穗病菌、水杨酸、茉莉酸甲酯、脱落酸胁迫下,SceIF5A均上调表达,推测SceIF5A基因在甘蔗响应黑穗病菌侵染过程中被诱导表达,且其表达受内源激素信号通路的调控. 本研究获得的SceIF5A基因的结构特征和功能作用模式为研究该基因在甘蔗与黑穗病菌互作中的作用积累了基础数据.
Eukaryotic translation initiation factor 5A (eIF5A) is ubiquitous in animals, plants and fungi. To reveal the responses of sugarcane eIF5A gene to biotic stresses, the present study first obtained an EST sequence with 93% homologs to Zea mays eIF5A (GenBank Accession Number:EU958725.1) from a suppression subtractive hybridization (SSH) library of sugarcane challenged by Sporisorium scitamineum. The EST sequence was then used as the probe for in silico cloning to obtain a putative cDNA sequence of sugarcane eIF5A gene. This gene was validated by reverse transcription-PCR (RT-PCR) amplification sequencing, which showed that the sequence cloned by in silico cloning was in accordance with the sequence amplified by RT-PCR method, and was named as SceIF5A (GenBank Accession Number: KJ577595). Bioinformatics analysis indicated that SceIF5A, with a length of 1 174 bp, containing a 483 bp open reading frame (ORF) encoding 160 amino acids protein, was a stable acidic cytoplasm protein with a weight of 17 453.6 Da containing 12 highly conserved amino acid sequences. Real-time quantitative PCR analysis showed that the expression of SceIF5A was up-regulated under smut fungus infection and the treatments of SA, MeJA and ABA. The results suggested that SceIF5A in sugarcane is most probably involved in response to sugarcane smut fungus infection and to hormone related bioprocess signaling. The results of this study regarding the structure and function of SceIF5A should lay the foundation for further research in the elaboration of functions of this gene during the interaction between sugarcane and S. scitamineum.


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国家自然科学基金项目(31101196)、福建省杰出青年科学基金项目(2015J06006)、福建省高等学校新世纪优秀人才支持计划(JA14095)和农业部甘蔗生物学与遗传育种重点实验室开放课题项目资助 Supported by the National Natural Science Foundation of China (31101196), the Natural Science Foundation of Fujian Province (2015J06qyx), the Program for New Century Excellent Talents in Fujian Province Universities (JA14095), and the Open Fund of Key Laboratory of Sugarcane Biology and Genetic Breeding, Ministry of Agriculture
更新日期/Last Update: 2016-01-05