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Selection and validation of reference genes for gene expression studies by reverse transcription quantitative PCR in Xanthomonas citri subsp. citri during infection of Citrus sinensis

机译:通过逆转录定量PCR在柠檬黄单胞菌亚种中进行基因表达研究的参考基因的选择和验证。柑橘感染期间的柠檬

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摘要

Xanthomonas citri subsp. citri (Xcc) causes citrus canker, a worldwide disease found mainly in sweet oranges (Citrus sinensis (L.) Osbeck). The expression of nine candidate internal reference genes was analyzed in Xcc grown alone and during C. sinensis infection to identify genes most suitable for comparative expression studies in Xcc using reverse transcription quantitative PCR (qRT-PCR). The stability of these genes was determined using the programs geNorm, NormFinder and BestKeeper. The genes most suitable for data normalization during C. sinensis infection were atpD, rpoB, gyrA and gyrB. The use of at least three reference genes is essential for accurate data normalization in Xcc.
机译:Xanthomonas citri subsp.。柠檬(Xcc)引起柑橘溃疡病,这是一种全世界性疾病,主要发现于甜橙(Citrus sinensis(L.)Osbeck)。分析了九种候选内部参考基因在单独生长的中华绒螯蟹中以及在中华绒毛线虫感染过程中的表达,从而确定了最适合使用逆转录定量PCR(qRT-PCR)在Xcc中进行比较表达研究的基因。这些基因的稳定性使用geNorm,NormFinder和BestKeeper程序确定。最适合在中华梭菌感染期间进行数据标准化的基因是atpD,rpoB,gyrA和gyrB。至少三个参考基因的使用对于Xcc中的准确数据标准化至关重要。

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