相黄单胞菌最佳内参基因选择及潜在靶基因鉴定。菊花感染。

IF 2 Q3 BIOCHEMICAL RESEARCH METHODS
Shu-Cheng Chuang, Shefali Dobhal, Teresita D Amore, Anne M Alvarez, Mohammad Arif
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引用次数: 0

摘要

相黄单胞菌。dieffenbachiae (Xpd)是天南星科红祖菜(Anthurium)细菌性枯萎病的致病因子,被列为EPPO A2检疫生物。虽然Xpd的全基因组已经测序,但红掌细菌性枯萎病(ABB)的分子机制仍然未知。在Xpd与红掌相互作用的初始阶段,选择合适的内参基因是获得准确可靠的基因表达谱的关键。采用反转录定量PCR (RT-qPCR)数据,采用best keeper、geNorm和delta Ct (ΔCt)三种统计方法评估4个内参基因的稳定性。其中,rpoD和gyrB基因在Xpd与敏感品种“Marian seefth”互作过程中表现出最一致的基因表达谱,而atpD和thyA基因在4个时间点(0、0.5、1和2 h)的表达谱不太稳定。通过使四个致病性相关基因的表达水平正常化,验证了这些内参候选基因的适用性。相互作用1小时后观察到编码黄原胶生物合成糖基转移酶的gumD的高度上调表达,表明它可能是xpd -红掌病理系统中关键的毒力决定因素。本研究确定的稳定内参基因将有助于今后在xpd -红掌病理系统中更准确、更全面的基因表达研究。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Optimal Reference Gene Selection and Potential Target Gene Identification During Xanthomonas phaseoli pv. dieffenbachiae-Anthurium andreanum Infection.

Xanthomonas phaseoli pv. dieffenbachiae (Xpd), the causal agent of bacterial blight in Anthurium within the Araceae family, is listed as an EPPO A2 quarantine organism. Although the whole genome of Xpd has been sequenced, the molecular mechanisms underlying anthurium bacterial blight (ABB) remain unknown. Selecting an optimal reference gene is crucial for obtaining accurate and reliable gene expression profiles during the initial interactions between Xpd and Anthurium. The stability of four reference genes was evaluated by applying three statistical methods-BestKeeper, geNorm, and delta Ct (ΔCt)-using reverse-transcription quantitative PCR (RT-qPCR) data. The rpoD and gyrB genes exhibited the most consistent gene expression profiles, whereas atpD and thyA were less stable at four time points (0, 0.5, 1, and 2 h) during the interactions between Xpd and susceptible A. andreanum cultivar 'Marian Seefurth.' The suitability of these reference gene candidates was validated by normalizing the gene expression levels of four pathogenicity-related genes. The highly upregulated expression of gumD, which encodes xanthan biosynthesis glycosyltransferase, observed after 1 h of interaction, suggests it may be a key virulence determinant in the Xpd-Anthurium pathosystem. The stable reference genes identified here will facilitate more accurate and comprehensive gene expression studies in the Xpd-Anthurium pathosystem going forward.

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来源期刊
Methods and Protocols
Methods and Protocols Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (miscellaneous)
CiteScore
3.60
自引率
0.00%
发文量
85
审稿时长
8 weeks
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