毛白杨翻译起始因子基因PtoeIF5A2的克隆及其表达特性分析

林业科学 ›› 2014, Vol. 50 ›› Issue (2): 63-69.

毛白杨翻译起始因子基因PtoeIF5A2的克隆及其表达特性分析

管阳, 王宏芝, 张杰伟, 陈亚娟, 朱丹, 丁莉萍, 魏建华

北京市农林科学院北京农业生物技术研究中心农业基因资源与生物技术北京市重点实验室北京 100097

收稿日期:2013-03-27 修回日期:2013-06-20 出版日期:2014-02-25 发布日期:2014-03-11
基金资助:
国家重点基础研究发展计划（973）项目（2012CB114501）；国家高技术研究发展计划（863）项目（2011AA100201）；国家自然科学基金项目（30972392）。

Isolation and Expression of Eukaryotic Translation Initiation Factor 5A2 in Populus tomentosa

Guan Yang, Wang Hongzhi, Zhang Jiewei, Chen Yajuan, Zhu Dan, Ding Liping, Wei Jianhua

Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Sciences Beijing Key Laboratory of Agricultural Genetic Resources and Biotechnology Beijing 100097

Received:2013-03-27 Revised:2013-06-20 Online:2014-02-25 Published:2014-03-11
Contact: 魏建华

摘要/Abstract

摘要：

真核翻译起始因子5A（eIF5A）在植物生长发育和抗逆过程中发挥着重要作用。以2个月的毛白杨‘BJHR01’幼苗总RNA反转录的cDNA为模板，克隆得到PtoeIF5A2 （GenBank登录号：HQ529480）基因全长cDNA序列。该基因包含483 bp开放读码框，编码160个氨基酸，预测蛋白分子质量为18 kDa，等电点为5.76。Real-Time qRT-PCR分析表明：PtoeIF5A2 在6个月毛白杨幼苗叶片中的表达量最高，约为其在幼根中表达量的5倍。PtoeIF5A2 响应了干旱、ABA（200 μmol ·L^-1）、低温（4 ℃），尤其是NaCl（300 mmol ·L^-1）胁迫。在NaCl胁迫24 h后，PtoeIF5A2 的表达量上调25倍，结合其启动子中含有6个病原体与高盐响应（GT-1 box）元件，推测该基因在响应高盐胁迫时起着重要作用。

关键词: 真核翻译起始因子5A, 毛白杨, 盐胁迫, PtoeIF5A2

Abstract:

Eukaryotic translation initiation factor 5A (eIF5A) plays an important role in plant development and various stresses. In this study, the full length PtoeIF5A2 (GenBank Accession number: HQ529480) cDNA was isolated from the cDNA library prepared from 2-month-old Populus tomentosa‘BJHR01’seedlings using RT-PCR technique. The PtoeIF5A2 coding region contained 483 nucleotides, encoding 160 amino acids with the molecular weight of about 18 kDa and PI of 5.76. The real time quantitative RT-PCR displayed that the expression of PtoeIF5A2 was highest in leaves in 6-month-old seedlings, about 5 times higher than that of in root. Moreover, the expression of PtoeIF5A2 in P. tomentosa responded to the drought, ABA and low temperature, especially NaCl treatment. After 24 h under NaCl, the PtoeIF5A2 expression was 25 times higher than control, and the PtoeIF5A2 promoter contained 6 response elements to pathogen and salinity, indicating that PtoeIF5A2 would be mainly involved in response of salt stress.

Key words: eIF5A, Populus tomentosa, salt stress, PtoeIF5A2

中图分类号:

S718.46

管阳, 王宏芝, 张杰伟, 陈亚娟, 朱丹, 丁莉萍, 魏建华. 毛白杨翻译起始因子基因PtoeIF5A2的克隆及其表达特性分析[J]. 林业科学, 2014, 50(2): 63-69.

Guan Yang, Wang Hongzhi, Zhang Jiewei, Chen Yajuan, Zhu Dan, Ding Liping, Wei Jianhua. Isolation and Expression of Eukaryotic Translation Initiation Factor 5A2 in Populus tomentosa[J]. Scientia Silvae Sinicae, 2014, 50(2): 63-69.

参考文献

付乾堂, 余迪求. 2010.拟南芥AtWRKY 25 、AtWRKY 26 和AtWRKY 33 在非生物胁迫条件下的表达分析.遗传,32(8): 848-856.
(Fu Q T, Yu D Q. 2010. Expression profiles of AtWRKY25,AtWRKY26 and AtWRKY33 under abiotic stresses. Hereditas, 32(8): 848-856. [in Chinese] )
靳宝锋,何昆,胡美茹,等. 2004.真核起始因子5A与细胞周期G1/S调控的研究.中国实验血液学杂志,(4): 325-328.
(Jin B F, He K, Hu M R, et al. 2004. The effect of eIF-5A on the G_1-S in cell cycle regulation. Journal of Experimental Hematology, (4): 325-328. [in Chinese] )
Byers T L, Lakanen J R,Coward J K, et al. 1994. The role of hypusine depletion in cytostasis induced by S-adenosyl-L-methionine decarboxylase inhibition: new evidence provided by 1-methylspermidine and 1, 12-dimethylspermine. Biochem J, 303: 363-368.
Cano V S, Jeon G A, Johansson H E, et al. 2008. Mutational analyses of human eIF5A-1 identification of amino acid residues critical for eIF5A activity and hypusine modification. FEBS J, 275(1): 44-58.
Chen J H, Xia X L, Yin W L. 2009. Expression profiling and functional characterization of a DREB2-type gene from Populus euphratica. Biochem Bioph Res Co, 378(3): 483-487.
Chen Z P, Yan Y P, Ding Q J, et al. 1996. Effects of inhibitors of deoxyhypusine synthase on the differentiation of mouse neuroblastoma and erythroleukemia cells. Cancer Lett, 105(2): 233-239.
Chou W C, Huang Y W, Tsay W S, et al. 2004. Expression of genes encoding the rice translation initiation factor, eIF5A, is involved in developmental and environmental responses. Plant Physiol, 121(1): 50-57.
Dias C A, Gregio A P, Rossi D, et al. 2012. EIF5A interacts functionally with eEF2. Amino Acids, 42(2/3): 697-702.
Duguay J, Jamal S, Liu Z, et al. 2007. Leaf specific suppression of deoxyhypusine synthase in Arabidopsis thaliana enhances growth without negative pleiotropic effects. J Plant Physiol, 164(4): 408-420.
Hopkins M T, Lampi Y, Wang T W, et al. 2008. Eukaryotic translation initiation factor 5A is involved in pathogen-induced cell death and development of disease symptoms in Arabidopsis. Plant Physiol, 148(1): 479-489.
Lee Y, Kim H K, Park H E, et al. 2002. Effect of N-l,7-diaminoheptane,an inhibitor of deoxyhypusine synthase, on endothelial cell growth, differentiation and apoptosis. Mol Cell Biochem, 237(1/2): 69-76.
Liu Z, Duguay J, Ma F, et al. 2008. Modulation of eIF5A1 expression alters xylem abundance in Arabidopsis thaliana. J Exp Bot, 59(4): 939-950.
Ma Y, Eriko M, Byung-Kook H, et al. 2010. Pumpkin eIF5A isoforms interact with components of the translational machinery in the cucurbit sieve tube system. Plant J, 64(3): 536-550.
Park H C, Kim M L, Kang Y H, et al. 2004. Pathogen- and NaCl-induced expression of the SCaM- 4 promoter is mediated in part by a GT-1 box that interacts with a GT-1-like transcription factor. Plant Physiol, 135(4): 2150-2161.
Thompson J E, Hopkin M T, Taylor C, et al. 2004. Regulation of senescence by eukaryotic translation initiation factor 5A implications for plant growth and development. Trends Plant Sci, 9(4): 174-179.
Tome M E, Gerner E W. 1996. Hypusine modification in eukaryotic initiation factor 5A in rodent cells selected for resistance to growth inhibition by ornithine decarboxylase-inhibiting drugs. Biochem J, 320: 55-60.
Wang L, Xu C, Wang C, et al. 2012. Characterization of a eukaryotic translation initiation factor 5A homolog from Tamarix androssowii involved in plant abiotic stress tolerance. BMC Plant Biology, 12(118): 1471-2229.
Xu J, Zhang B, Jiang C, et al. 2011. RceIF5A, encoding an eukaryotic translation initiation factor 5A in Rosa chinensis, can enhance thermotolerance, oxidative and osmotic stress resistance of Arabidopsis thaliana. Plant Mol Biol, 75(1/2): 167-178.

[1]	刘道凤, 王霞, 代银, 杨建峰, 马婧, 李名扬, 眭顺照. 蜡梅转录因子CpTAF10基因的克隆及功能分析[J]. 林业科学, 2019, 55(6): 176-183.
[2]	朱嘉磊, 薄慧娟, 李璇, 文春燕, 王江, 聂立水, 田菊, 宋莲君. 不同毛白杨无性系林分蓄积量的长期水氮耦合效应[J]. 林业科学, 2019, 55(5): 27-35.
[3]	安胜男, 马晓军, 朱礼智. 木粉增强P34HB生物复合材料的制备及其结构性能表征[J]. 林业科学, 2019, 55(3): 125-133.
[4]	江成, 周厚君, 赵岩秋, 何辉, 楚立威, 宋学勤, 卢孟柱. 干旱和高盐胁迫下钙离子依赖核酸酶基因CDD对银腺杨84K生长发育的影响[J]. 林业科学, 2019, 55(2): 33-40.
[5]	李建波, 贾会霞, 张进, 刘伯斌, 胡建军, 王丽娟, 卢孟柱. 毛白杨PtoWOX4a基因过表达对次生生长的影响[J]. 林业科学, 2018, 54(2): 52-59.
[6]	李豆豆, 席本野, 唐连峰, 冯超, 贺曰林, 张亚雄, 刘龙龙, 刘金强, 贾黎明. 砂壤土下滴灌毛白杨幼林土壤水分运移规律与模拟[J]. 林业科学, 2018, 54(12): 157-168.
[7]	席本野, 王烨, 贾黎明. 滴灌施肥下施氮量和施氮频率对毛白杨生物量及氮吸收的影响[J]. 林业科学, 2017, 53(5): 63-73.
[8]	王凯利, 傅鹰, 周明兵. 毛竹SQUAMOSA启动子结合蛋白SBP家族基因的鉴定及表达模式分析[J]. 林业科学, 2017, 53(12): 50-61.
[9]	储文渊, 王玉娇, 朱东悦, 陈竹, 严涵薇, 项艳. 盐和干旱胁迫下杨树新内参基因的筛选[J]. 林业科学, 2017, 53(10): 70-79.
[10]	张秀英, 宋瑞清, 张星耀. 杨树受溃疡病菌感染后转录因子的应答变化[J]. 林业科学, 2015, 51(4): 110-115.
[11]	邱念伟, 周峰, 张士超, 杨东, 刘媛, 宋贤慧, 郑媛. 人工模拟倒春寒对杨树叶片活力的影响[J]. 林业科学, 2014, 50(7): 17-22.
[12]	宋曰钦, 谢宗强, 翟明普, 贾黎明. 三倍体毛白杨不同有机残体分解及氮磷释放特征[J]. 林业科学, 2014, 50(4): 1-7.
[13]	杨秀艳, 张华新, 张丽, 刘正祥, 杨升, 武香. NaCl胁迫对唐古特白刺幼苗生长及离子吸收、运输与分配的影响[J]. 林业科学, 2013, 49(9): 165-171.
[14]	马金龙, 姜国斌, 姚善泾, 金华. 盐胁迫下杨树质外体离子对气体交换参数的影响[J]. 林业科学, 2013, 49(7): 40-47.
[15]	蒋淑磊;陈加飞;赵树堂;卢孟柱;. 应用nanoLC-MS/MS分析毛白杨次生维管系统的蛋白质表达谱[J]. , 2013, 49(5): 43-53.