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在不同实验条件下，用于西伯利亚偃麦草（）实时荧光定量 PCR 基因表达分析的合适参考基因的选择。

Selection of Suitable Reference Genes for RT-qPCR Gene Expression Analysis in Siberian Wild Rye () under Different Experimental Conditions.

机构信息

The State Key Laboratory of Grassland Agro-Ecosystems, Key Laboratory of Grassland Livestock Industry Innovation, Ministry of Agriculture and Rural Affairs, College of Pastoral Agriculture Science and Technology, Lanzhou University, Lanzhou 730020, China.

出版信息

Genes (Basel). 2019 Jun 13;10(6):451. doi: 10.3390/genes10060451.

DOI:10.3390/genes10060451
PMID:31200580
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6627066/
Abstract

, which is a perennial and self-pollinated grass, is the typical species of the genus , which plays an important role in forage production and ecological restoration. No reports have, so far, systematically described the selection of optimal reference genes for reverse transcriptase quantitative real-time polymerase chain reaction (RT-qPCR) analysis in . The goals of this study were to evaluate the expression stability of 13 candidate reference genes in different experimental conditions, and to determine the appropriate reference genes for gene expression analysis in . Five methods including Delta Ct (ΔCt), BestKeeper, NormFinder, geNorm, and RefFinder were used to assess the expression stability of 13 potential reference genes. The results of the RefFinder analysis showed that and were the most stable reference genes in different genotypes. and had the most stable expression in different developmental stages. and were suitable reference genes in different tissues. Under salt stress, and were identified as the most stable reference genes. and showed the most stability under heat stress. For cold stress, and presented the highest degree of expression stability. and were considered as the most stable reference genes under osmotic stress. The optimal reference genes were selected to investigate the expression pattern of target gene in different conditions. This study provides suitable reference genes for further gene expression analysis using RT-qPCR in .

摘要

是一种多年生自花授粉的草本植物，是属的典型物种，在饲料生产和生态恢复中起着重要作用。到目前为止，还没有系统地描述过在 中用于逆转录定量实时聚合酶链反应 (RT-qPCR) 分析的最佳参考基因的选择。本研究的目的是评估 13 个候选参考基因在不同实验条件下的表达稳定性，并确定用于 基因表达分析的合适参考基因。使用 Delta Ct (ΔCt)、BestKeeper、NormFinder、geNorm 和 RefFinder 五种方法评估 13 个潜在参考基因的表达稳定性。RefFinder 分析的结果表明，在不同基因型下， 和 是最稳定的参考基因。在不同发育阶段， 和 具有最稳定的表达。在不同组织中， 和 是合适的参考基因。在盐胁迫下， 和 被鉴定为最稳定的参考基因。在热胁迫下， 和 表现出最稳定的表达。在冷胁迫下， 和 表现出最高的表达稳定性。在渗透胁迫下， 和 被认为是最稳定的参考基因。选择最佳的参考基因来研究不同条件下靶基因 的表达模式。本研究为进一步使用 RT-qPCR 在 中进行基因表达分析提供了合适的参考基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cb1/6627066/d2d674c718b4/genes-10-00451-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cb1/6627066/ddb64821002c/genes-10-00451-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cb1/6627066/1cabc19c7fbe/genes-10-00451-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cb1/6627066/6fafbe761e2a/genes-10-00451-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cb1/6627066/ad91089fa6b1/genes-10-00451-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cb1/6627066/1c3b90125e3b/genes-10-00451-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cb1/6627066/1cec2a88bf3f/genes-10-00451-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cb1/6627066/a53acb6f91d9/genes-10-00451-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cb1/6627066/d2d674c718b4/genes-10-00451-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cb1/6627066/ddb64821002c/genes-10-00451-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cb1/6627066/1cabc19c7fbe/genes-10-00451-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cb1/6627066/6fafbe761e2a/genes-10-00451-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cb1/6627066/ad91089fa6b1/genes-10-00451-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cb1/6627066/1c3b90125e3b/genes-10-00451-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cb1/6627066/1cec2a88bf3f/genes-10-00451-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cb1/6627066/a53acb6f91d9/genes-10-00451-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0cb1/6627066/d2d674c718b4/genes-10-00451-g008.jpg

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