[1]代红军,秦晨亮,徐伟荣.赤霞珠葡萄发育后期RT-PCR内参基因的筛选和验证[J].江苏农业学报,2016,(03):668.[doi:10.3969/j.issn.1000-4440.2016.03.028]
　DAI Hong-jun,QIN Chen-liang,XU Wei-rong.Screening and validation of reference genes for real-time fluorescence quantitative PCR during grape berry development of Cabernet Sauvignon[J].,2016,(03):668.[doi:10.3969/j.issn.1000-4440.2016.03.028]
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赤霞珠葡萄发育后期RT-PCR内参基因的筛选和验证()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2016年03期
页码:
668
栏目:
园 艺
出版日期:
2016-06-30

文章信息/Info

Title:
Screening and validation of reference genes for real-time fluorescence quantitative PCR during grape berry development of Cabernet Sauvignon
作者:
代红军1; 2;  秦晨亮1; 2;  徐伟荣1; 2
1.宁夏大学农学院,宁夏 银川 750021; 2.宁夏大学教育部葡萄与葡萄酒工程中心,宁夏 银川 750021
Author(s):
DAI Hong-jun1; 2;  QIN Chen-liang1; 2;  XU Wei-rong1; 2
1.School of Agriculture, Yinchuan 750021, China; 2.Grape and Wine Engineering Research Center of Ningxia University, Ministry of Education, Yinchuan 750021, China
关键词:
赤霞珠葡萄;  内参基因;  实时荧光定量PCR
Keywords:
Cabernet Sauvignon grape;  reference gene;  qRT-PCR
分类号:
S663.1
DOI:
10.3969/j.issn.1000-4440.2016.03.028
文献标志码:
A
摘要:
葡萄果实发育后期基因表达水平的变化对果实品质的形成具有重要影响,选择合适的内参基因是提高实时荧光定量PCR分析准确性的首要条件。本试验以赤霞珠葡萄发育后期不同取样时间的果皮为材料,通过qRT-PCR分析了常用候选内参基因β-actin、EF-1α、GAPDH和SAND的表达变化,借助geNorm、NormFinder和BestKeeper程序筛选出在葡萄发育后期qRT-PCR分析的理想内参基因。结果表明, β-actin的稳定性最好,其次是SAND,而GAPDH和EF-1α的稳定性相对较低; 同时,用筛选的内参基因β-actin和SAND分析葡萄白藜芦醇合成途径中二苯乙烯合酶基因的表达水平,其表达规律一致。说明, β-actin和SAND是研究赤霞珠葡萄发育后期基因表达的理想内参基因。
Abstract:
The change of gene expression level during berry ripening is crucial for the formation of grape fruit quality. The selection of a suitable reference gene is an important factor for accurate gene expression analysis by real-time fluorescence quantitative PCR(qRT-PCR). In this study, the pericarp of wine grape(Cabernet Sauvignon)was taken as material at different time during berry ripening to detect the expression levels of four commonly used housekeeping genes β-actin, EF-1α, GAPDH and SAND by qRT-PCR, so as to select reliable reference genes using softwares geNorm, NormFinder, and BestKeeper. The stability was found to the best in β-actin, followed by SAND. The expression of STS gene in the pathway of grape resveratrol anthocyanin biosynthesis was analyzed, and the results indicated that the variation dynamics of STS using β-actin as reference gene was exactly consistent with that using SAND as reference gene. Therefore, β-actin and SAND are ideal reference genes for gene expression level analysis during grape berry development.

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备注/Memo

备注/Memo:
收稿日期:2015-10-25
基金项目:国家自然科学基金项目(31260456)
作者简介::代红军(1967-),女,宁夏银川人,博士,教授,研究方向为葡萄栽培生理。(Tel)13139589616;(E-mail)dai_hj@nxu.edu.cn
更新日期/Last Update: 2016-06-30