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Selection and evaluation of suitable reference genes for quantitative gene expression analysis during infection of Cucumis sativus with Pectobacterium brasiliense.
Yuan, Lifang; Zhao, Yurong; Xie, Hua; Shi, Yanxia; Xie, Xuewen; Chai, Ali; Li, Lei; Li, Baoju.
Afiliação
  • Yuan L; Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, China.
  • Zhao Y; Shandong Academy of Grapes, Shandong Academy of Agricultural Sciences, Shandong, China.
  • Xie H; Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, China.
  • Shi Y; Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Sciences, Beijing Key Laboratory of Agricultural Genetic Resources and Biotechnology, Beijing, China.
  • Xie X; Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, China.
  • Chai A; Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, China.
  • Li L; Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, China.
  • Li B; Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, China.
J Appl Microbiol ; 132(5): 3717-3734, 2022 May.
Article em En | MEDLINE | ID: mdl-35138009
AIMS: Bacterial soft rot caused by Pectobacterium brasiliense (Pbr) has resulted in severe economic losses of cucumber production in northern China. Quantitative reverse transcription PCR (RT-qPCR) is widely used to determine the fold change in the expression of genes of interest, and an appropriate reference gene played a critical role in the evaluation of genes expression. However, the suitable reference genes for transcript normalization during the interaction between cucumber and Pbr have not yet been systematically validated. In this study, we aimed to identify the suitable reference genes for accurate and reliable normalization of cucumber and Pbr RT-qPCR data. METHODS AND RESULTS: We selected 14 candidate reference genes for cucumber and 10 candidate reference genes for Pbr were analysed by using four algorithms (the deltaCt method, BestKeeper, NormFinder and geNorm). Furthermore, five genes in cucumber involved in plant resistance and five genes in Pbr related to the virulence were selected to confirm the reliability of the reference genes by RT-qPCR. CsARF (ADP-ribosylation factor 1) and pgi (glucose-6-phosphate isomerase) were suggested as the most suitable reference genes for cucumber and Pbr respectively. CONCLUSION: Our results suggested that CsARF (ADP-ribosylation factor 1) and pgi (glucose-6-phosphate isomerase) could be as the reference genes to normalize expression data for cucumber and Pbr during the process of pathogen-host interaction respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first systematic study of the optimal reference genes specific to cucumber and Pbr, which could help advance the molecular interactions research in Cucurbitaceae vegetables and Pectobacterium species pathosystems.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Cucumis sativus / Pectobacterium Tipo de estudo: Prognostic_studies País/Região como assunto: America do sul / Brasil Idioma: En Revista: J Appl Microbiol Assunto da revista: MICROBIOLOGIA Ano de publicação: 2022 Tipo de documento: Article País de afiliação: China País de publicação: Reino Unido
Texto completo
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Cucumis sativus / Pectobacterium Tipo de estudo: Prognostic_studies País/Região como assunto: America do sul / Brasil Idioma: En Revista: J Appl Microbiol Assunto da revista: MICROBIOLOGIA Ano de publicação: 2022 Tipo de documento: Article País de afiliação: China País de publicação: Reino Unido