Selection of reliable reference genes for RT-qPCR during methyl jasmonate, salicylic acid and hydrogen peroxide treatments in Ganoderma lucidum

Lu, Xiao-Xiao; Liu, Yongzhi; Zhao, Linchao; Liu, Yong-Nan; Ren, Ang

doi:10.1007/s11274-018-2476-x

Cited by 19 publications

(16 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Several studies have been performed on the selection and evaluation of stable reference genes under different conditions, such as different development stages and nutrient conditions as well as abiotic stresses in Morchella [19], Ganoderma lucidum [20,21,22], Volvariella volvacea [23,24], and L. edodes [25,26]. Nevertheless, the information about the reference genes used for qRT-PCR normalization of the hot-air-dried L. deodes is rather limited.…”

Section: Introductionmentioning

confidence: 99%

Selection of Reference Genes for qRT-PCR Analysis in Lentinula edodes after Hot-Air Drying

et al. 2018

View full text Add to dashboard Cite

Volatile sulfur compounds gradually develop in Lentinula edodes after hot-air drying, and many genes are involved in the generation of these sulfur compounds. The expression stability of reference genes may vary in a particular experimental treatment when analyzing their expressions by quantitative real-time polymerase chain reaction (qRT-PCR). In this study, the expression profile of 17 candidate genes was assessed in L. edodes under treatment at 50 °C for 0, 1, 2, and 3 h, and the expression stability of each reference gene was analyzed by three statistical algorithms, including geNorm, NormFinder, and BestKeeper. Results indicated that the two optimal reference genes for mycelium and fruiting body were CAC and DAHP as well as CAC and NUP, respectively. Additionally, CAC and DAHP were found to be the two most stable reference genes across the mycelium and fruiting body set. Our results will provide a genetic foundation for further research on the metabolism genes of sulfur compounds in L. edodes.

show abstract

Section: Introductionmentioning

confidence: 99%

Selection of Reference Genes for qRT-PCR Analysis in Lentinula edodes after Hot-Air Drying

et al. 2018

View full text Add to dashboard Cite

show abstract

“…RNA isolation and cDNA synthesis were performed as previously described method [19]. Briefly, 0.2 g mycelia were collected and subsequently were disrupted under liquid nitrogen conditions.…”

Section: Rna Isolation and Cdna Synthesismentioning

confidence: 99%

Analysis of reference genes stability and histidine kinase expression under cold stress in Cordyceps militaris

Liu

et al. 2020

PLoS ONE

View full text Add to dashboard Cite

The development of fungal fruiting bodies from a hyphal thallus is inducible under low temperature (cold stress). The molecular mechanism has been subject to surprisingly few studies. Analysis of gene expression level has become an important means to study gene function and its regulation mechanism. But identification of reference genes (RGs) stability under cold stress have not been reported in famous medicinal mushroom-forming fungi Cordyceps militaris. Herein, 12 candidate RGs had been systematically validated under cold stress in C. militaris. Three different algorithms, geNorm, NormFinder and BestKeeper were applied to evaluate the expression stability of the RGs. Our results showed that UBC and UBQ were the most stable RGs for cold treatments in short and long periods, respectively. 2 RGs (UBC and PP2A) and 3 RGs (UBQ, TUB and CYP) were the suitable RGs for cold treatments in short and long periods, respectively. Moreover, target genes, two-componentsystem histidine kinase genes, were selected to validate the most and least stable RGs under cold treatment, which indicated that use of unstable expressed genes as RGs leads to biased results. Our results provide a good starting point for accurate reverse transcriptase quantitative polymerase chain reaction normalization by using UBC and UBQ in C. militaris under cold stress and better support for understanding the mechanism of response to cold stress and fruiting body formation in C. militaris and other mushroom-forming fungi in future research.

show abstract

“…RNA isolation and cDNA synthesis were performed as previously described method [41]. Briefly, 0.2 g mycelia were collected and subsequently were disrupted under liquid nitrogen conditions.…”

Section: Rna Isolation and Cdna Synthesismentioning

confidence: 99%

Analysis of Reference Genes stability and Histidine Kinase Expression under Cold Stress in Cordyceps militaris

Liu

et al. 2020

Preprint

View full text Add to dashboard Cite

Background: The development of fungal fruiting bodies from a hyphal thallus represents a transition from simple to complex multicellularity that is inducible under low temperature (cold stress). The molecular mechanism has been subject to surprisingly few studies. Analysis of gene expression level has become an important means to study gene function and its regulation mechanism. But identification of reference genes (RGs) stability under cold stress have not been reported in famous medicinal mushroom-forming fungi Cordyceps militaris.Results: Herein, 12 candidate RGs had been systematically validated under cold stress in C. militaris.Three different algorithms, geNorm, NormFinder and BestKeeper were applied to evaluate the expression stability of the RGs. Our results showed that UBC and UBQ were the most stable RGs for cold treatments in short and long time, respectively. 2 RGs (UBC and PP2A) and 3 RGs (UBQ, TUB and CYP) were the suitable housekeeping genes for cold treatments in short and long time, respectively.Moreover, target genes, two-component-system histidine kinase genes, were selected to validate the most and least stable RGs under cold treatment, which indicated that use of unstable expressed genes as housekeeping genes leads to biased results. Conclusions:Our results provide a good starting point for accurate reverse transcriptase quantitative polymerase chain reaction normalization by using UBC and UBQ in C. militaris under cold stress and better support for understanding the mechanism of response to cold stress and fruiting body formation in C. militaris and other mushroom-forming fungi in future research. BackgroundCordyceps militaris, a famous traditional Chinese medicine, has been used as a healthy food for a long time in China. The C. militaris fruiting body has anti-inflammatory [1], anti-tumor [2], antiinfluenza virus [3] and radio-protection [4] functions. Methods for commercial production of fruiting bodies of this fungus have been established in artificial media [5,6] or with insects, such as silkworm Bombyx mori pupae [7]. The molecular mechanism of mushroom fruiting body formation is a basic biological problem. The essential role of light in fruit body development in C. militaris was demonstrated in previous study [8]. A blue-light receptor gene white collar-1 (wc-1) inactivation

show abstract

Selection of reliable reference genes for RT-qPCR during methyl jasmonate, salicylic acid and hydrogen peroxide treatments in Ganoderma lucidum

Cited by 19 publications

References 47 publications

Selection of Reference Genes for qRT-PCR Analysis in Lentinula edodes after Hot-Air Drying

Selection of Reference Genes for qRT-PCR Analysis in Lentinula edodes after Hot-Air Drying

Analysis of reference genes stability and histidine kinase expression under cold stress in Cordyceps militaris

Analysis of Reference Genes stability and Histidine Kinase Expression under Cold Stress in Cordyceps militaris

Contact Info

Product

Resources

About