[1]张青,陆明星,祝树德.灰飞虱2种热激蛋白基因Hsp70的克隆、分析[J].江苏农业学报,2015,(06):1257-1264.[doi:doi:10.3969/j.issn.1000-4440.2015.06.010]
　ZHANG Qing,LU Ming-xing,ZHU Shu-de.Cloning,sequence analysis and expression profiling of two heat shock protein 70 genes in small brown plant hopper,Laodelphax striatellus (Hemiptera:Delphacidae)[J].,2015,(06):1257-1264.[doi:doi:10.3969/j.issn.1000-4440.2015.06.010]
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灰飞虱2种热激蛋白基因Hsp70的克隆、分析()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年06期
页码:
1257-1264
栏目:
植物保护
出版日期:
2015-12-31

文章信息/Info

Title:
Cloning,sequence analysis and expression profiling of two heat shock protein 70 genes in small brown plant hopper,Laodelphax striatellus (Hemiptera:Delphacidae)
作者:
张青12; 陆明星1; 祝树德1
（1.扬州大学园艺与植物保护学院，江苏扬州225009；2.苏州市农业科学院，江苏苏州215155）
Author(s):
ZHANG Qing12; LU Ming-xing1; ZHU Shu-de1
(1.School of Horticulture and Plant Protection,Yangzhou University,Yangzhou 225009,China; 2.Suzhou Academy of Agricultural Sciences,Suzhou 215155,China)
关键词:
灰飞虱; 热激蛋白70; 发育; 温度; 实时定量PCR
Keywords:
Laodelphax striatellus; heat shock protein 70; development; temperature; real-time quantitative PCR
分类号:
S435.112+.3
DOI:
doi:10.3969/j.issn.1000-4440.2015.06.010
文献标志码:
A
摘要:
灰飞虱[Laodelphax striatellus (Fallén)]是一种分布广，对环境有很强适应性的重要水稻害虫。热激蛋白70（Hsp70）家族是与生物体环境适应最为密切的一类蛋白质。本研究通过RT-PCR与RACE技术克隆灰飞虱Hsp70基因，利用生物信息方法分析了该热激蛋白的特征，采用实时荧光定量PCR技术研究该基因在不同生长发育阶段和不同温度条件下表达变化规律。结果显示:克隆了Hsp70基因的2种cDNA全长序列，分别命名为LsHsc70（GenBank登录号为KF660252）和LsHsp70（GenBank登录号为KF660251）。LsHsc70全长为2 399 bp，编码657个氨基酸，编码的蛋白质等电点为5.3，分子量为73 000；LsHsp70全长为2 468 bp，编码690个氨基酸，编码的蛋白质等电点为5.8，分子量为74 900。LsHsc70和LsHsp70的氨基酸序列中均含有 Hsp70蛋白质家族的3个签名序列及1个ATP-GTP结合位点。系统进化关系分析结果表明它们与多种昆虫的Hsp70氨基酸序列有较高的同源性。不同发育阶段的灰飞虱体内LsHsc70和LsHsp70呈现出不同的表达模式，LsHsc70在雄性成虫阶段表达量达到最大值，而LsHsp70的最高表达量在1龄若虫阶段。不同性别的灰飞虱成虫体内LsHsc70表达量有显著差异，而LsHsp70没有显著差异。高温和低温胁迫都可以诱导灰飞虱体内LsHsc70和LsHsp70的表达，在-4 ℃和-9 ℃的低温胁迫下，LsHsc70和LsHsp70分别达最大的表达量。在40 ℃，LsHsc70和LsHsp70表达水平均在处理后0.5 h时最高。而在-4 ℃，LsHsc70和LsHsp70的表达水平均在处理后1 h时达到最大值。结果表明灰飞虱可以通过调节体内的LsHsc70和LsHsp70表达，来应对不良的环境温度。
Abstract:
The small brown planthopper Laodelphax striatellus (Fallén),an important rice pest insect,feeds on rice and causes the decrease of yield and quality.The small brown planthopper is widely distributed and adapts to various environments.Heat shock protein 70 subfamily is considered as the protein related mostly to environmental adaption of organism.In this study,two full-length cDNA encoding Hsp70,LsHsc70 (GenBank accession no.KF660252) and LsHsp70 (GenBank accession no.KF660251)， were cloned. The complete cDNA of LsHsc70 was 2 399 bp in length,encoding a protein of 657 amino acids,with the theoretical isoelectric point of 5.3 and molecular weight of 73 000.The complete cDNA of LsHsp70 was 2 468 bp in length,encoding a protein of 690 amino acids,with the theoretical isoelectric point of 5.8 and molecular weight of 74 900.The amino acid sequence of LsHsc70 and LsHsp70 both contained three signature sequences of Hsp70 family and a ATP-GTP binding site.The phylogenic tree demonstrated that LsHsc70 and LsHsp70 shared high homology with Hsp70 amino acid sequence from other insect species.Real-time PCR analyses revealed that the expression level of LsHsc70 differed from LsHsp70 in different developmental stages;LsHsc70 reached the highest level in the male adult of L.striatellus,while LsHsp70 expression level peaked in the first instar nymphs. LsHsc70 showed significant differences between male and female adults,but LsHsp70 did not.The LsHsc70 and LsHsp70 expressions of L.striatellus could be induced by both heat and coldness stress.The highest LsHsc70 and LsHsp70 levels were observed at -4 ℃ and -9 ℃,respectively.At 40 ℃,LsHsc70 and LsHsp70 levels reached the peaks 0.5 h after treatment.It was indicated that L.striatellus could regulate the levels of LsHsc70 and LsHsp70 to adapt to adverse temperatures.

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备注/Memo

备注/Memo:
收稿日期：2015-09-08 基金项目：国家科技支撑计划项目(2011BAD16B03); 苏州市科技支撑计划项目(SNG201204) 作者简介：张青（1969-），男， 江苏高邮人， 博士， 副研究员，研究方向为农业昆虫与害虫治理。（Tel）0512-66700877;（E-mail）zhangqingsz@163.com 通讯作者：祝树德，（E-mail）sdzhu@yzu.edu.cn
更新日期/Last Update: 2015-12-31