Suppr超能文献

通过定量实时PCR筛选绿豆在生物胁迫和激素处理下的内参基因

Screening of Reference Genes under Biotic Stress and Hormone Treatment of Mung Bean () by Quantitative Real-Time PCR.

作者信息

Zhou Yanyan, Liu Huan, Wu Ting, Zheng Yu, Wang Ruimin, Xue Dong, Yan Qiang, Yuan Xingxing, Chen Xin

机构信息

College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China.

Institute of Industrial Crops, Jiangsu Academy of Agricultural Sciences/Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement, Nanjing 210014, China.

出版信息

Genes (Basel). 2023 Aug 30;14(9):1739. doi: 10.3390/genes14091739.

Abstract

Mung bean () production has been greatly threatened by numerous diseases. Infection with these pathogens causes extensive changes in gene expression and the activation of hormone signal transduction. Quantitative real-time PCR (qRT-PCR) is the most common technique used for gene expression validation. Screening proper reference genes for mung bean under pathogen infection and hormone treatment is a prerequisite for ensuring the accuracy of qRT-PCR data in mung bean disease-resistance research. In this study, six candidate reference genes (, , , , , and ) were selected to evaluate the expression stability under four soil-borne disease pathogens (, , , and ) and five hormone treatments (SA, MeJA, ETH, ABA, and GA). In the samples from different treatments, the Ct value distribution of the six candidate reference genes was different. Under the condition of hormone treatment, the Ct value ranged from a minimum of 17.87 for to a maximum of 29.63 for . Under the condition of pathogen infection, the Ct value ranged from a minimum of 19.43 for to a maximum of 31.82 for . After primer specificity analysis, it was found that was not specific, so the five reference genes , , , and were used in subsequent experiments. The software products GeNorm, NormFinder, BestKeeper and RefFinder were used for qRT-PCR data analysis. In general, the best candidates reference genes were: for SA, ABA, GA3, and treatment; for ETH treatment; for MeJA and treatment; and for and treatment. The most stably expressed genes in all samples were , while was the least stable reference gene. Finally, the reliability of the reference gene was further validated by analysis of the expression profiles of four mung bean genes (, , , and ) selected from transcriptome data. Our results provide more accurate information for the normalization of qRT-PCR data in mung bean response to pathogen interaction.

摘要

绿豆()的生产受到多种病害的严重威胁。这些病原体的感染会导致基因表达发生广泛变化以及激素信号转导的激活。定量实时聚合酶链反应(qRT-PCR)是用于基因表达验证的最常用技术。在病原体感染和激素处理条件下筛选合适的绿豆内参基因是确保绿豆抗病性研究中qRT-PCR数据准确性的前提。本研究选择了6个候选内参基因(、、、、和),以评估在4种土传病害病原体(、、、和)和5种激素处理(SA、MeJA、ETH、ABA和GA)下的表达稳定性。在不同处理的样本中,6个候选内参基因的Ct值分布不同。在激素处理条件下,Ct值范围从的最小值17.87到的最大值29.63。在病原体感染条件下,Ct值范围从的最小值19.43到的最大值31.82。经过引物特异性分析,发现不具有特异性,因此在后续实验中使用了5个内参基因、、、和。使用GeNorm、NormFinder、BestKeeper和RefFinder软件产品进行qRT-PCR数据分析。总体而言,最佳候选内参基因是:SA、ABA、GA3和处理时为;ETH处理时为;MeJA和处理时为;和处理时为。所有样本中表达最稳定的基因是,而最不稳定的内参基因是。最后,通过对从转录组数据中选择的4个绿豆基因(、、、和)的表达谱分析,进一步验证了内参基因的可靠性。我们的结果为绿豆对病原体相互作用反应中qRT-PCR数据的标准化提供了更准确的信息。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/306e/10530681/5d2bfd3a5381/genes-14-01739-g001a.jpg

相似文献

2
Signs and symptoms to determine if a patient presenting in primary care or hospital outpatient settings has COVID-19.
Cochrane Database Syst Rev. 2022 May 20;5(5):CD013665. doi: 10.1002/14651858.CD013665.pub3.
3
Systemic pharmacological treatments for chronic plaque psoriasis: a network meta-analysis.
Cochrane Database Syst Rev. 2021 Apr 19;4(4):CD011535. doi: 10.1002/14651858.CD011535.pub4.
4
Diagnostic test accuracy and cost-effectiveness of tests for codeletion of chromosomal arms 1p and 19q in people with glioma.
Cochrane Database Syst Rev. 2022 Mar 2;3(3):CD013387. doi: 10.1002/14651858.CD013387.pub2.
5
Thoracic imaging tests for the diagnosis of COVID-19.
Cochrane Database Syst Rev. 2022 May 16;5(5):CD013639. doi: 10.1002/14651858.CD013639.pub5.
8
Antibody tests for identification of current and past infection with SARS-CoV-2.
Cochrane Database Syst Rev. 2022 Nov 17;11(11):CD013652. doi: 10.1002/14651858.CD013652.pub2.
9
Antidepressants for pain management in adults with chronic pain: a network meta-analysis.
Health Technol Assess. 2024 Oct;28(62):1-155. doi: 10.3310/MKRT2948.
10
Rapid, point-of-care antigen tests for diagnosis of SARS-CoV-2 infection.
Cochrane Database Syst Rev. 2022 Jul 22;7(7):CD013705. doi: 10.1002/14651858.CD013705.pub3.

引用本文的文献

本文引用的文献

1
2
SlWRKY30 and SlWRKY81 synergistically modulate tomato immunity to by directly regulating .
Hortic Res. 2023 Mar 15;10(5):uhad050. doi: 10.1093/hr/uhad050. eCollection 2023 May.
3
4
Global distribution, traditional and modern detection, diagnostic, and management approaches of associated with legume crops.
Front Microbiol. 2023 Feb 6;13:1091288. doi: 10.3389/fmicb.2022.1091288. eCollection 2022.
5
6
Endophytic antagonize soil-borne fungal pathogens and suppress wilt complex disease in chickpea plants ( L.).
Front Microbiol. 2022 Nov 2;13:994847. doi: 10.3389/fmicb.2022.994847. eCollection 2022.
7
The Phytophthora effector Avh94 manipulates host jasmonic acid signaling to promote infection.
J Integr Plant Biol. 2022 Nov;64(11):2199-2210. doi: 10.1111/jipb.13358. Epub 2022 Oct 14.
8
Selection and Validation of Reference Genes for qRT-PCR Gene Expression Analysis in .
Genes (Basel). 2022 Aug 14;13(8):1445. doi: 10.3390/genes13081445.
9
Analysis of var Gene Transcript Patterns by Quantitative Real-Time PCR.
Methods Mol Biol. 2022;2470:149-171. doi: 10.1007/978-1-0716-2189-9_13.
10
Identification and Validation of Reliable Reference Genes for Gene Expression Studies in .
Genes (Basel). 2022 Apr 19;13(5):714. doi: 10.3390/genes13050714.

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验