| 张金平1,2,3、何晓明2,3、江彪2,3、谢大森2,3、彭庆务2、王永飞1、马三梅1.镰刀菌酸胁迫下节瓜实时荧光定量
PCR分析内参基因的选择[J].广东农业科学,2014,41(10):120-125 |
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镰刀菌酸胁迫下节瓜实时荧光定量
PCR分析内参基因的选择 |
| Reference gene selection for real-time quantitativePCR normalization under fusaric acid stress inBenincasa hispida Cogn. var. chieh-qua How |
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| DOI： |
| 中文关键词: 节瓜 实时荧光定量PCR 内参基因 镰刀菌酸胁迫 |
| 英文关键词: enincasa hispida Cogn. var. chieh-qua How real-time quantitative PCR reference genes fusaric acid
stress |
| 基金项目:广东省科技攻关项目（2012A020100006）；广东
省自然科学基金（S2013010012888）；广东省试验室体系建设开
放运行费(粤财教[2013]112 号) ；广东省科技计划项目(粤财教
[2011]510 号) |
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| 摘要点击次数: 1469 |
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| 中文摘要: |
| 针对不同试验材料及条件选择合适的内参基因是正确利用实时荧光定量PCR 分析目的基因表达模式
的前提条件。以4 个常用看家基因（GAPDH尧Actin尧UBQ尧CYP）和2个新内参基因（CACS尧F-box）作为候选内参基因、
运用实时荧光定量PCR 技术、分析它们在节瓜不同组织（根尧茎尧叶）及镰刀菌酸（fusaric acid, FA）胁迫下的表达稳定
性。经RefFinder 在线软件综合分析、结果表明、节瓜经枯萎病毒素FA 胁迫后Actin 和F-box 表达稳定；而在节瓜不
同组织中F-box 基因表达稳定性最高；另外、传统常用看家基因GAPDH 和CYP因在不同试验条件下总体表达水平
都比较差而不适合作为相应条件下的内参基因。研究结果为节瓜抗枯萎病相关基因的表达分析奠定了基础。 |
| 英文摘要: |
| Appropriate reference genes for specific experimental materials and conditions are critical for analyzing the
expression of target gene by real-time quantitative PCR. In this study, four common housekeeping genes GAPDH, Actin,
UBQ, CYP and two novel reference genes CACS, F-box were chosen for identifying the expression stability in Benincasa
hispida Cogn. var. chieh-qua How samples from various plant tissues (roots, stems and leaves) or under fusaric acid (FA)
stress. The expression stability of candidate genes was determined and ranked using RefFinder software. The results
revealed a remarkably stable expression pattern of Actin and F-box when samples were stressed by fusaric acid. F-box
gene showed the most stable expression level in different tissues. Additionally, expression of traditional housekeeping genes
GAPDH and CYP showed great variations and therefore they were considered unsuitable as reference genes. This study has
laid the foundation for the study of resistance-related genes expression for fusarium wilt in B. hispida Cogn. var. chieh-qua
How |
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