| 温度和药剂胁迫下茶刺盲蝽内参基因稳定性分析 |
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| 引用本文： | 王政,朱茜,孟倩倩,孙世伟,高圣风,刘爱勤.温度和药剂胁迫下茶刺盲蝽内参基因稳定性分析[J].植物保护学报,2019,46(3):530-541. |
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| 作者姓名： | 王政 朱茜 孟倩倩 孙世伟 高圣风 刘爱勤 |
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| 作者单位： | 中国热带农业科学院香料饮料研究所, 海南省热带香辛饮料作物遗传改良与品质调控重点实验室, 万宁 571533,中国热带农业科学院香料饮料研究所, 海南省热带香辛饮料作物遗传改良与品质调控重点实验室, 万宁 571533;海南大学热带农林学院, 海口 570228,中国热带农业科学院香料饮料研究所, 海南省热带香辛饮料作物遗传改良与品质调控重点实验室, 万宁 571533,中国热带农业科学院香料饮料研究所, 海南省热带香辛饮料作物遗传改良与品质调控重点实验室, 万宁 571533,中国热带农业科学院香料饮料研究所, 海南省热带香辛饮料作物遗传改良与品质调控重点实验室, 万宁 571533,中国热带农业科学院香料饮料研究所, 海南省热带香辛饮料作物遗传改良与品质调控重点实验室, 万宁 571533;海南大学热带农林学院, 海口 570228 |
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| 基金项目： | 海南省自然科学基金（317272），中国热带农业科学院基本科研业务费专项（1630142018016） |
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| 摘 要： | 为筛选特定条件下实时荧光定量PCR检测体系中能稳定表达的茶刺盲蝽Helopeltis theivora内参基因,从其转录组中筛选并克隆11个候选基因Actin、β-tubulin1、RPL13A、EF1α、RPS3A、GAPDH、18S RNA、G6PDH、EIF4A、TBP和UBQ,测定它们在不同温度及药剂胁迫下mRNA的表达水平,并通过geNorm、NormFinder、BestKeeper、Delta Ct和RefFinder软件(算法)分析各基因的稳定性。结果表明,11个候选基因引物均具有良好的特异性和扩增效率(90.10%～96.87%)。温度胁迫下,EIF4A有最小的平均变异度(mean variability,M)、稳定值(stability value,SV)和几何平均值,分别为0.200、0.096和2.060;RPS3A的平均标准偏差(average of standard deviation,STDEV)最小,为0.605;二者是表达最稳定的基因。药剂胁迫下,RPL13A的M值和STDEV最小,分别为0.123和0.660;RPS3A的SV、标准偏差(standard deviation,SD)和几何平均值均最小,分别为0.063、0.336和1.189;二者是表达最稳定的基因;Actin为最不稳定基因。全样品评价中,RPL13A(M值为0.282、几何平均值为1.565)和RPS3A(SV为0.099、STDEV为0.614)的评价值同比最小,是表达最稳定的基因;最不稳定的基因为Actin、β-tubulin1和TBP。geNorm分析结果还显示所有处理条件下最适内参基因数目均为2个。表明EIF4A和RPS3A可作为茶刺盲蝽与温度相关的抗性基因表达研究的内参基因,RPS3A和RPL13A可作为该虫抗药基因表达研究的内参基因。
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| 关 键 词： | 茶刺盲蝽 温度 药剂 内参基因稳定性 实时荧光定量PCR |
| 收稿时间： | 2018/10/24 0:00:00 |
Stability evaluation of reference genes in tea mosquito bug Helopeltis theivora under temperature and pesticide stress |
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| Wang Zheng,Zhu Xi,Meng Qianqian,Sun Shiwei,Gao Shengfeng and Liu Aiqin.Stability evaluation of reference genes in tea mosquito bug Helopeltis theivora under temperature and pesticide stress[J].Acta Phytophylacica Sinica,2019,46(3):530-541. |
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| Authors: | Wang Zheng Zhu Xi Meng Qianqian Sun Shiwei Gao Shengfeng and Liu Aiqin |
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| Institution: | Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops, Research Institute of Spice and Beverage, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, Hainan Province, China,Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops, Research Institute of Spice and Beverage, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, Hainan Province, China;Institute of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, Hainan Province, China,Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops, Research Institute of Spice and Beverage, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, Hainan Province, China,Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops, Research Institute of Spice and Beverage, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, Hainan Province, China,Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops, Research Institute of Spice and Beverage, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, Hainan Province, China and Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops, Research Institute of Spice and Beverage, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, Hainan Province, China;Institute of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, Hainan Province, China |
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| Abstract: | |
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| Keywords: | Helopeltis theivora temperature pesticide expression stability of reference genes quantitative real-time PCR |
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