Screening and analysis on reference genes of <i>Ganoderma pseudoferreum</i>

ZHAO Xinyang; LUO Youhong; CAI Haibin; ZHOU Yi; TU Min

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ZHAO X Y, LUO Y H, CAI H B, et al. Screening and analysis on reference genes of Ganoderma pseudoferreum [J]. Fujian Journal of Agricultural Sciences，2024，39（5）：1−9

Citation:

ZHAO X Y, LUO Y H, CAI H B, et al. Screening and analysis on reference genes of Ganoderma pseudoferreum [J]. Fujian Journal of Agricultural Sciences，2024，39（5）：1−9

ZHAO X Y, LUO Y H, CAI H B, et al. Screening and analysis on reference genes of Ganoderma pseudoferreum [J]. Fujian Journal of Agricultural Sciences，2024，39（5）：1−9

Citation:

ZHAO X Y, LUO Y H, CAI H B, et al. Screening and analysis on reference genes of Ganoderma pseudoferreum [J]. Fujian Journal of Agricultural Sciences，2024，39（5）：1−9

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Screening and analysis on reference genes of Ganoderma pseudoferreum

ZHAO Xinyang^{1, 2
,},
LUO Youhong^{1, 2},
CAI Haibin²,
ZHOU Yi^{1
,
,},
TU Min²

1.
College of Agriculture, Yangtze University, Jingzhou, Hubei,　434000, China
2.
Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan　571101, China

Received Date: 2024-02-27
Rev Recd Date: 2024-04-24

Available Online: 2024-06-26

Abstract

Abstract

: Objective The stability of candidate reference genes of Ganoderma pseudoferreum under different stress treatments were evaluated to provide reference for exploring the gene function and molecular mechanism of G.pseudoferreum infestation of rubber tree roots. Methods The G. pseudoferreum mycelia under five abiotic stresses ( temperature, salt, oxidation, pH, drought ) and one biotic stress (biocontrol bacteria) were collected to isolate RNA and reverse transcribed into cDNA. Real-time fluorescence quantitative PCR technology was used to amplify the six candidate reference genes (UBC, ACT, RPL6, β-TUB, APT, 28s), and the softwares of geNorm, NormFinder, BestKeeper and RefFinder were used to evaluate genes stability to find the most suitable reference genes for different stresses. Results The RNA of all samples showed two clear bands, and the melting curves of the six candidate reference genes showed obvious single peak. Combined analysis of geNorm, NormFinder, Bestkeeper and RefFinder, found that the stability of gene expression under temperature stress was UBC ＞ ACT ＞ RPL6 ＞ β-TUB ＞ 28s ＞ APT, and the stability of gene expression under salt stress was ACT ＞ RPL6 ＞ UBC ＞ APT ＞ β-TUB ＞ 28s; the stability of gene expression under oxidative stress was UBC ＞ ACT ＞ 28s ＞ APT ＞ β-TUB ＞ RPL6; the stability of gene expression under pH stress was RPL6 ＞ UBC ＞ APT ＞ ACT ＞ β-TUB ＞ 28s, the stability of gene expression under drought stress was ACT ＞ UBC ＞ β-TUB ＞ RPL6 ＞ APT ＞ 28s, and the stability of gene expression under biotic stress was UBC ＞ ACT ＞ RPL6 ＞ 28s ＞ APT ＞ β-TUB. Conclusion Combining all genes and experimental conditions, we recommend genes ACT and UBC as the best reference genes for normalizing gene expression under drought, oxidation, temperature and biotic stress experimental conditions. Genes ACT and RPL6 are the most suitable reference genes under salt stress, and UBC and RPL6 are the most suitable reference genes under pH stress.
- Rubber tree,
- Red root rot disease,
- Ganoderma pseudoferreum,
- Reference genes,
- RT-qPCR

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References(28)

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