JOURNAL OF MICROBIOLOGICAL METHODS  卷: 114   页: 62-65   出版年: JUL 2015

关键词:Clonostachys rosea; Reference gene; Elongation factor; Mycoparasitism; Sclerotinia sclerotiorum; Real-time PCR

摘要

Reference genes are important to precisely quantify gene expression by real-time PCR. In order to identify stable and reliable expressed genes in mycoparasite Clonostachys rosea indifferent modes of nutrition, seven commonly used housekeeping genes, 18S rRNA, actin, beta-tubulin, elongation factor 1, ubiquitin, ubiquitin-conjugating enzyme and glyceraldehyde-3-phosphate dehydrogenase, from the effective biocontrol isolate C rosea 67-1 were tested for their expression under sclerotial induction and during vegetative growth on PDA medium. Analysis by three software programs showed that differences existed among the candidates. Elongation factor 1 was most stable; the M value in geNorm, SD value in Bestkeeper and stability value in Normfinder analysis were 0.405, 0.450 and 0.442, respectively, indicating that the gene elongation factor 1 could be used to normalize gene expression in C. rosea in both vegetative growth and parasitic process. By using elongation factor 1, the expression of a serine protease gene, sep, in different conditions was assessed, which was consistent with the transcriptomic data. This research provides an effective method to quantitate expression changes of target genes in C rosea, and will assist in further investigation of parasitism-related genes of this fungus. (C) 2015 Elsevier B.V. All rights reserved.