摘要
【目的】昆虫在高温或农药的胁迫下,通过高效表达热休克蛋白(HSP)等建立应激自我保护机制。本研究为从转录组水平上认识大豆蚜Aphis glycines在热应激和吡虫啉胁迫下hsp70和hsc70 mRNA表达分子机制,进而寻找自我保护应激反应中的薄弱环节,为大豆蚜的生物防治提供理论基础。【方法】采用同源克隆、RACE技术和实时荧光定量PCR等方法研究不同热激时间和热激后不同恢复时间及不同吡虫啉浓度对大豆蚜4龄若虫hsp70和hsc70的表达影响。【结果】37℃热激后,大豆蚜4龄若虫中hsp70表达量先上调,1 h时升至对照组的10.36倍(P<0.05),然后逐渐下降。同样热激后恢复时间的长短对大豆蚜若蚜中hsp70的表达具有显著影响。热激处理后,大豆蚜若蚜中hsp70立即大量表达,表达量为对照组的8.78倍(P<0.05),随后表达量下降至对照组水平,而hsc70的表达量并没有显著变化(P>0.05)。大豆蚜若蚜受吡虫啉的胁迫时,其hsp70和hsc70的表达量受吡虫啉的浓度及胁迫的时间的影响,呈现先升高后下降的趋势,具有明显的短期效应。【结论】吡虫啉诱导大豆蚜hsp70和hsc70表达量的上调;而热胁迫对hsp70和hsc70 mRNA具有不同的表达模式,高温可以诱导hsp70的表达,但对hsc70没有明显的诱导作用。
[ Aim] Insects will express heat shock proteins （HSPs） efficiently to establish self-protection mechanism under high temperature and pesticide stress. This study aims to know the molecular mechanisms of hsp70 and hsc70 mRNA expression in Aphis glycines under thermal and imidacloprid stress at the transcriptome level and look for the vulnerable spot, so to provide some theoretical basis for biological control. [ Methods ] We adopted the homology-based cloning, RACE technique and real-time fluorescent quantitative PCR methods to research the expression of hspTO and hsc70 in the 4th instar nymphs of A. glycines at different heat shock time or different recovery time after heat shock and after treatment with different concentrations of imidacloprid. [ Results] The hsp70 transcription level in the 4th instar nymphs of A. glycines increased first after heat shock at 37℃, reaching 10.36-fold as high as that of the control （P 〈 0.05） at 1 h after heat shock, and then declined little by little. The recovery time after heat shock significantly influenced the hsp70 expression quantity. The hsp70 of A. glycines was abundantly expressed immediately after heat shock, with the expression level of 8.78-fold as high as that of the control （P 〈 0.05 ）, and then decreased to the control level, while the hsc70 expression quantity did not change significantly （P 〉 0.05 ）. Treated with imidacloprid, the peak time of hsp70 and hsc70 expression quantity in the 4th instar nymphs of A. glycines varied with the concentration of imidacloprid and stress time, showing an obvious short-term effect with a downward trend after rising first.[ Conclusion ] The increases. However, expression patterns ： effect on hsc70. transcription level of hsp70 and hsc70 in A. glycines under imidacloprid stress hsp70 and hsc70 in A. glycines subjected to heat shock have different transcription the high temperature induces the expression of hsp70, but has no obvious inductiveeffect on hsc70.
出处
《昆虫学报》
CAS
CSCD
北大核心
2014年第4期387-394,共8页
Acta Entomologica Sinica
基金
公益性行业(农业)科研专项(201103002)
国家级大学生创新训练项目
黑龙江省博士后基金(IBH-Z09261)
关键词
大豆蚜
热休克蛋白
吡虫啉
热激
MRNA表达
Aphis glycines
heat shock protein （HSP）
imidacloprid
heat shock
mRNA expression
