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Differential coexistence of multiple genotypes of Ophiocordyceps sinensis in the stromata, ascocarps and ascospores of natural Cordyceps sinensis

Yu-Ling Li, Xiu-Zhang Li, Yi-Sang Yao, Zi-Mei Wu, Ling Gao, Ning-Zhi Tan, Zhou-Qing Luo, Wei-Dong Xie, Jian-Yong Wu, View ORCID ProfileJia-Shi Zhu
doi: https://doi.org/10.1101/2022.06.20.496766
Yu-Ling Li
1State Key Laboratory of Plateau Ecology and Agriculture, Qinghai Academy of Animal and Veterinary Sciences, Qinghai University, Xining, Qinghai, China
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Xiu-Zhang Li
1State Key Laboratory of Plateau Ecology and Agriculture, Qinghai Academy of Animal and Veterinary Sciences, Qinghai University, Xining, Qinghai, China
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Yi-Sang Yao
2Shenzhen Key Laboratory of Health Science and Technology, Institute of Biopharmaceutical and Health Engineering, Shenzhen International Graduate School, Tsinghua University, Shenzhen 518055, China
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Zi-Mei Wu
3School of Life Sciences, Tsinghua University, Beijing, China
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Ling Gao
2Shenzhen Key Laboratory of Health Science and Technology, Institute of Biopharmaceutical and Health Engineering, Shenzhen International Graduate School, Tsinghua University, Shenzhen 518055, China
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Ning-Zhi Tan
2Shenzhen Key Laboratory of Health Science and Technology, Institute of Biopharmaceutical and Health Engineering, Shenzhen International Graduate School, Tsinghua University, Shenzhen 518055, China
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Zhou-Qing Luo
3School of Life Sciences, Tsinghua University, Beijing, China
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Wei-Dong Xie
2Shenzhen Key Laboratory of Health Science and Technology, Institute of Biopharmaceutical and Health Engineering, Shenzhen International Graduate School, Tsinghua University, Shenzhen 518055, China
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Jian-Yong Wu
4State Key Laboratory of Chinese Medicine and Molecular Pharmacology, Shenzhen 518057, Guangdong, China
5Department of Applied Biology and Chemistry Technology, The Hong Kong Polytechnic University, Hong Kong
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Jia-Shi Zhu
1State Key Laboratory of Plateau Ecology and Agriculture, Qinghai Academy of Animal and Veterinary Sciences, Qinghai University, Xining, Qinghai, China
2Shenzhen Key Laboratory of Health Science and Technology, Institute of Biopharmaceutical and Health Engineering, Shenzhen International Graduate School, Tsinghua University, Shenzhen 518055, China
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  • ORCID record for Jia-Shi Zhu
  • For correspondence: zhujosh@163.com
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Abstract

Objective To examine the differential occurrence of Ophiocordyceps sinensis genotypes in the stroma, stromal fertile portion (SFP), and ascospores of natural Cordyceps sinensis.

Methods Immature and mature C. sinensis were harvested. Mature C. sinensis were continuously cultivated in our laboratory (altitude 2,200 m). The SFPs (with ascocarps) and ascospores of C. sinensis were collected for microscopic and molecular analyses using species-/genotype-specific primers.

Results Fully and semi-ejected ascospores were collected from the same specimens. The semi-ejected ascospores tightly adhered to the surface of the asci. The multicellular heterokaryotic ascospores showed uneven staining of nuclei. The ascospores were found to contain several genotypes of O. sinensis, Samsoniella hepiali, and an AB067719-type fungus. The genotypes within AT-biased Cluster-A occurred in all compartments of C. sinensis, but those within AT-biased Cluster-B were absent in the ascospores. Genotypes #13‒14 of O. sinensis occurred differentially in fully and semi-ejected ascospores, which featured DNA segment substitutions and genetic material recombination between the genomes of the parental fungi, Hirsutella sinensis and the AB067719-type fungus. These offspring genotypes combined with varying abundances of S. hepiali in the 2 types of ascospores participated in the control of the development, maturation and ejection of the ascospores.

Conclusion Multiple genotypes of O. sinensis coexist differentially in the SFP and 2 types of C. sinensis ascospores, along with S. hepiali and the AB067719-type fungus. The fungal components in different combinations and their dynamic alterations in the compartments of C. sinensis during maturation play symbiotic roles in the lifecycle of natural C. sinensis.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted June 20, 2022.
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Differential coexistence of multiple genotypes of Ophiocordyceps sinensis in the stromata, ascocarps and ascospores of natural Cordyceps sinensis
Yu-Ling Li, Xiu-Zhang Li, Yi-Sang Yao, Zi-Mei Wu, Ling Gao, Ning-Zhi Tan, Zhou-Qing Luo, Wei-Dong Xie, Jian-Yong Wu, Jia-Shi Zhu
bioRxiv 2022.06.20.496766; doi: https://doi.org/10.1101/2022.06.20.496766
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Differential coexistence of multiple genotypes of Ophiocordyceps sinensis in the stromata, ascocarps and ascospores of natural Cordyceps sinensis
Yu-Ling Li, Xiu-Zhang Li, Yi-Sang Yao, Zi-Mei Wu, Ling Gao, Ning-Zhi Tan, Zhou-Qing Luo, Wei-Dong Xie, Jian-Yong Wu, Jia-Shi Zhu
bioRxiv 2022.06.20.496766; doi: https://doi.org/10.1101/2022.06.20.496766

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