文献类型：期刊文献

英文题名：Selection of suitable reference genes for quantitative real time PCR in different Tulasnella isolates and orchid-fungus symbiotic germination system

作者：Wang, Tao[1,2] Cao, Xiaolu[1] Wang, Xiaojing[1] Chi, Miao[1,2] Li, Lubin[1] Yao, Na[1]

第一作者：Wang, Tao

通信作者：Yao, N[1]

机构：[1]Chinese Acad Forestry, State Key Lab Tree Genet & Breeding, Key Lab Tree Breeding & Cultivat, Natl Forestry & Grassland Adm,Res Inst Forestry, Beijing 100091, Peoples R China;[2]Beijing Floriculture Engn Technol Res Ctr, Beijing Lab Urban & Rural Ecol Environm, Beijing Bot Garden, Beijing 100093, Peoples R China

年份：2021

卷号：48

期号：1

起止页码：527-538

外文期刊名：MOLECULAR BIOLOGY REPORTS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000604518700005)】；

基金：This work was supported by grants from National Natural Science Foundation of China (No. 31700547), and the Beijing Botanical Garden (No. BZ201901).

语种：英文

外文关键词：Reference genes; RT-qPCR; Tulasnella; Orchid mycorrhizal fungi

摘要：Under natural conditions, mycorrhizal symbiosis accompanies nearly the entire life cycle of orchids from seed germination through to flowering and fruiting. Tulasnella-like orchid mycorrhizal fungi are the most common mycorrhizal fungi found in association with orchid species. Presently suitable reference genes have not been systematically selected for the quantification of gene expression via Real-Time Quantitative Reverse Transcription PCR (RT-qPCR). We evaluated 12 candidate Tulasnella genes in nine different Tulasnella isolates and in the Dendrobium-fungal symbiotic germination associations followed by statistical analysis using the programs Bestkeeper, geNorm, and Normfinder to analyze the expression stability of the individual genes. The results showed that the EF2, UBC, and PP2A genes had the highest rankings with relatively stable expression levels across the different genotypes and during the symbiotic seed germination process by the three programs, and may be suitable for RT-qPCR normalization. Furthermore, the gene encoding C-5 Sterol desaturase (C5SD) was selected to verify the reliability of EF2, UBC, and PP2A expression during the Tulasnella-Dendrobium symbiotic seed germination process. This study is the first systematic exploration of optimal reference genes for gene expression studies during the colonization of orchid seeds by the mycorrhizal fungus Tulasnella.