AUTHOR=Ji Tingting , Ma Si , Liang Meiting , Wang Xueyun , Gao Lihong , Tian Yongqiang 

TITLE=Reference genes identification for qRT-PCR normalization of gene expression analysis in Cucumis sativus under Meloidogyne incognita infection and Pseudomonas treatment

JOURNAL=Frontiers in Plant Science

VOLUME=13

YEAR=2022

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2022.1061921

DOI=10.3389/fpls.2022.1061921

ISSN=1664-462X

ABSTRACT=<p>qRT-PCR is a common and key technical means to study gene expression in biological research. However, reliability and accuracy of quantification by qRT-PCR is entirely dependent on the identification of appropriate reference genes. Cucumber as an economical vegetable is widely cultivated worldwide and is subject to serious nematode infection, especially from <italic>M. incognita</italic>. Plant could employ beneficial soil bacteria in the rhizosphere to enhance plant adaptability to various stresses. In this study, the optimal reference genes in cucumber under <italic>M. incognita</italic> stress and <italic>Pseudomonas</italic> treatment were calculated and confirmed. A total of thirteen candidate reference genes were identified across three different treatments. Of these, geNorm, NormFinder and BestKeeper programs combined RefFinder software identified <italic>EF1</italic> and <italic>UBI</italic> are the most suitable reference gene in the root knot and whole root of cucumber infected <italic>M. incognita</italic>, respectively, and <italic>CACS</italic> is the most suitable reference gene in the whole root of cucumber treated by <italic>Pseudomonas</italic>. The work first validated the most suitable reference genes for the normalization gene expression in cucumber by nematode infected or <italic>Pseudomonas</italic> inoculated, and these results would facilitate the further research on <italic>M. incognita</italic> or <italic>Pseudomonas</italic> soil rhizosphere microbe interaction with cucumber.</p>