The Fvclp1 gene regulates mycelial growth and fruiting body development in edible mushroom Flammulina velutipes

Lyu, Xiaomeng; Jiang, Siyuan; Wang, Li; Chou, Tiansheng; Qingji, Wang; Li, Meng; Mukhtar, Irum; Xie, Baogui; Wang, Wei

doi:10.1007/s00203-021-02514-0

Cited by 8 publications

(10 citation statements)

References 24 publications

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“…In Cordyceps militaris , the best reference genes for inducing fruiting bodies in wheat medium and pupae are gpd / rpb1 and rpb1 / try , respectively [ 32 ]. However, during the fruiting bodies development of Flammulina velutipes , Botrytis cinerea and Agaricus bisporus , the stable reference genes are gapdh , tubulin and EF1-α , respectively [ 33 – 35 ]. In the research related to the development of O .…”

Section: Discussionmentioning

confidence: 99%

Stable reference gene selection for Ophiocordyceps sinensis gene expression studies under different developmental stages and light-induced conditions

Tong

Wei

Mao³

et al. 2023

PLoS ONE

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The molecular mechanism of Chinese cordyceps formation has received a substantial amount of attention because of its usage as traditional Chinese medicine. The formation process of Chinese cordyceps includes two parts: asexual proliferation (Ophiocordyceps sinensis proliferates in the hemolymph of Thitarodes armoricanus larvae) and sexual development (formation and development of fruiting bodies). Therefore, validation of reference genes under different development stages and experimental conditions is crucial for RT-qPCR analysis. However, there is no report on stable reference genes at the development stage of O. sinensis fruiting body. In this study, 10 candidate reference genes, Actin, Cox5, Tef1, Ubi, 18s, Gpd, Rpb1, Try, Tub1 and Tub2, were selected and calculated their expression stability using four methods: geNorm, NormFinder, BestKeeper, and Comparative △Ct. After comprehensive analysis of the results of these four methods with RefFinder, we determined that the most stable reference genes during asexual reproduction of O. sinensis were Tef1 and Tub1, while the most stable reference genes during fruiting body development were Tyr and Cox5, and the most stable reference genes under light-induced conditions were Tyr and Tef1. Our study provides a guidance for reference genes selections at different proliferation processes with light stress of O. sinensis, and represents a foundation for studying the molecular mechanism of Chinese cordyceps formation.

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Section: Discussionmentioning

confidence: 99%

Stable reference gene selection for Ophiocordyceps sinensis gene expression studies under different developmental stages and light-induced conditions

Tong

Wei

Mao³

et al. 2023

PLoS ONE

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“…5,6,15 The proteins encoded by Hd1 and Hd2 genes can interact to form a functional transcription factor but only if they are encoded by different alleles from a corresponding sublocus. 4,6 Many genes are involved in the mating type pathways in basidiomycetes, and the function of some of these genes, like the clampless1 (Clp1) gene, 16 has been established to a certain extent. It is known that Clp1 is an accessory gene involved in homeodomain (HD) genes in the matA pathway.…”

Section: ■ Introductionmentioning

confidence: 99%

“…Many genes are involved in the mating type pathways in basidiomycetes, and the function of some of these genes, like the clampless1 ( Clp1 ) gene, has been established to a certain extent. It is known that Clp1 is an accessory gene involved in homeodomain (HD) genes in the matA pathway.…”

Section: Introductionmentioning

confidence: 99%

Homeodomain 1 Genes of the Different HD Subloci of Flammulina velutipes Can Activate the HD Pathway and Are Involved in Mating, Clamp Cell Formation, and Upregulation of FvClp1

Liu,

Chou,

Wang

et al. 2024

J. Agric. Food Chem.

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Flammulina velutipes has two independent and functional mating type factors, HD and PR. The HD locus contains two separate subloci: HD-a and HD-b. In this study, we investigated the roles of Hd1 genes of the HD-a and HD-b subloci in the process of mating, clamp cell formation, and regulation of FvClp1 (F. velutipes clampless1 gene) gene expression in F. velutipes. To this end, we introduced Hd1 genes from mating compatible strains into F. velutipes monokaryon L11. Overexpression of Hd1 gene FvHd-a1-1 of the HD-a sublocus resulted in the formation of pseudoclamps in L11 monokaryons. L11 mutants overexpressing the Hd1 gene FvHd-b1-2 of the HD-b sublocus also similarly developed pseudoclamps in the L11 monokaryons. Moreover, these mutant L11 monokaryons produced complete clamps when crossed with monokaryotic strains that differed at the PR loci, i.e., when selective activation of the PR pathway was obtained through crossing. Thus, Hd1 genes of the two different HD subloci in F. velutipes can activate the HD mating type pathway and induce clamp cell formation. In addition, activation of the HD pathway resulted in upregulation of the FvClp1 gene. Finally, to complete clamp cell formation, activation of the PR pathway appears to be essential. Overall, these findings were beneficial for deepening our understanding of sexual reproduction and fruiting body development of edible fungi.

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“…Since then, studies have continued to show that A. tumefaciens- mediated transfer (ATMT) enables the random transfer of T-DNA into the genomes of fungi, including ascomycetes, basidiomycete and zygomycete ( Poyedinok and Blume, 2018 ). In recent years, Cordyceps militaris ( Zheng et al, 2011 ), Agaricus bisporus ( Wang et al, 2019 ), Flammulina velutipes ( Lyu et al, 2021 ) and other species have been successfully tested for A. tumefaciens -mediated transfer. However, this method has the disadvantage that it is not yet stable and consistent.…”

Section: Introductionmentioning

confidence: 99%

Construction of a heat-resistant strain of Lentinus edodes by fungal Hsp20 protein overexpression and genetic transformation

Ling

Zhao

2022

Front. Microbiol.

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The shiitake mushroom (Lentinus edodes) is the second most popular edible mushroom globally due to its rich nutritional value and health benefits associated with consumption. However, the characteristics of growing at low temperatures limit the area and time of its cultivating. We selected a low-temperature cultivar as the original strain. We proposed to construct a heat-shock protein expression vector to achieve genetic transformation in this low-temperature strain to improve the survivability of the strain against the heat-shock response. In this study, an overexpression vector pEHg-gdp-hsp20 for the heat shock protein 20 gene of A. bisporus was constructed using a homologous recombination method. This vector was transferred into dikaryotic and monokaryotic mycelia by the Agrobacterium tumefaciens-method. The integration of hygb and hsp20 into the genome of L. edodes mycelia was verified by growth experiments on resistant plates and PCR analysis. The expression of the reporter gene mgfp5 was verified by fluorescence microscopy analysis and statistically resulted in 18.52 and 26.39% positivity for dikaryon, and monokaryon, respectively. Real-time PCR analysis showed that the expression of the hsp20 gene was more than 10-fold up-regulated in the three transformants; the mycelia of the three overexpression transformants could resume growth after 24 h heat treatment at 40°C, but the mycelia of the starting strain L087 could not recover growth at 25°C indicating that strains that successfully expressed hsp20 had greater overall recovery after heat shock. According to the study, A. bisporus hsp20 gene overexpression effectively improves the defensive capability of low-temperature mushroom strains against heat shock, laying the foundation for breeding heat-resistant high-quality transgenic shiitake mushrooms.

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The Fvclp1 gene regulates mycelial growth and fruiting body development in edible mushroom Flammulina velutipes

Cited by 8 publications

References 24 publications

Stable reference gene selection for Ophiocordyceps sinensis gene expression studies under different developmental stages and light-induced conditions

Stable reference gene selection for Ophiocordyceps sinensis gene expression studies under different developmental stages and light-induced conditions

Homeodomain 1 Genes of the Different HD Subloci of Flammulina velutipes Can Activate the HD Pathway and Are Involved in Mating, Clamp Cell Formation, and Upregulation of FvClp1

Construction of a heat-resistant strain of Lentinus edodes by fungal Hsp20 protein overexpression and genetic transformation

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