Journal of Mushroom (한국버섯학회지)

The Korean Society of Mushroom Science (한국버섯학회)
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Detection of single-nucleotide polymorphism in RPB2 of Wolfiporia hoelen strains and assessment of its applicability for strain breeding

복령 균주의 RPB2 유전자 내 단일염기다형성 및 육종 활용성 분석

  • Su Yeon, Kim (Forest Microbiology Division, Department of Forest Bio-Resources, National Institute of Forest Science) ;
  • Mi-Jeong, Park (Forest Microbiology Division, Department of Forest Bio-Resources, National Institute of Forest Science) ;
  • Seong Hwan, Kim (Department of Microbiology, College of Science & Technology, Dankook University) ;
  • Kang-Hyeon, Ka (Forest Microbiology Division, Department of Forest Bio-Resources, National Institute of Forest Science)
  • 김수연 (국립산림과학원 산림생명자원연구부 산림미생물연구과) ;
  • 박미정 (국립산림과학원 산림생명자원연구부 산림미생물연구과) ;
  • 김성환 (단국대학교 과학기술대학 미생물학과) ;
  • 가강현 (국립산림과학원 산림생명자원연구부 산림미생물연구과)
  • Received : 2022.11.30
  • Accepted : 2022.12.22
  • Published : 2022.12.31
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Abstract

The demand for novel strains has been rising in the domestic market to increase the production of sclerotia from Wolfiporia hoelen. To improve strain breeding efficiency, we investigated whether single-nucleotide polymorphisms (SNPs) in the RNA polymerase II subunit (RPB2) gene, which may be linked to the mating type locus, are useful for distinguishing monokaryons from dikaryons in Korean W. hoelen strains. We designed a specific primer set to efficiently amplify a region of RPB2 using PCR with the genomic DNA of 12 cultivated strains and 31 wild strains of W. hoelen collected from Korea. Nucleotide sequences of the PCR-amplified RPB2 genes were determined and analyzed for the presence of SNPs among the 43 W. hoelen strains. Previously reported SNP loci were detected in the RPB2 gene of all W. hoelen strains tested. However, these previously reported SNP loci could not be applied to differentiate monokaryons from dikaryons in approximately one-third of Korean wild strains with homozygous genotypes. Three additional SNPs in the RPB2 gene, which may improve the ability to distinguish monokaryons from dikaryons, were identified by searching through the multiple sequence alignments of the 43 W. hoelen strains. The applicability of these three novel SNPs, together with the previously known SNPs, in the RPB2 gene to W. hoelen strain breeding was verified by examining the hybrid strains and their parental strains.

국내 시장에서는 Wolfiporia hoelen의 균핵 생산량을 늘리기 위해 새로운 균주 육종을 요구되고 있다. 이에 본 연구는 국내에서 수집한 31개의 야생 균주와 12개의 재배 균주에 대해, 최근 보고된 교배형 연관 유전자의 RPB2의 단일염기다형성(single nucleotide polymorphism, SNP)을 적용해 단핵균사와 이핵균사를 구분할 수 있는지 확인함으로써 SNP 정보가 육종에 유용한지 알아 보고자 수행되었다. 이를 위해 해당 정보를 효율적으로 얻을 수 있는 프라이머도 개발하였다. 분석 결과, 국내 야생 균주들은 동형접합인 경우가 중국 균주보다 많아 기존 SNP의 한계를 확인하였다. 이를 보완하고자 RPB2 유전자에서 3개의 SNP를 추가로 발견하였으며, 이를 통해 단핵균사와 이핵균사의 구분 능력을 높였다. 나아가 4개의 SNP를 기존에 육성한 교잡균주와 교잡에 사용한 단포자 균주에 적용함으로써 육종에서의 활용 가능성이 있음을 확인하였다.

Keywords

Acknowledgement

본 연구는 국립산림과학원 (FP0800-2020-02)과제의 예산을 지원받아 수행되었으며 이에 감사드립니다.

References

  1. Au CH, Wong MC, Bao D, Zhang M, Song C, Song W, Law PTW, Kues U, Kwan HS. 2014. The genetic structure of the A mating-type locus of Lentinula edodes. Gene 535: 184-190. https://doi.org/10.1016/j.gene.2013.11.036
  2. Igari N, Minoura S, Okada M. 1995. Multinucleate nature, and mating by use of isozyme analysis in Poria cocos. Mycoscience 36: 405-411. https://doi.org/10.1007/BF02268624
  3. Jang YA, Lee JT. 2015. The evaluation of antioxidant, antiinflammatory, and anti-aging of extract solvent and Poria cocos by parts. Kor J Aesthet Cosmetol 13: 377-383.
  4. Jo WS, Lee SH, Koo J, Ryu S, Kang MG, Lim SY, Park SC. 2017. Morphological characteristics of fruit bodies and basidiospores of Wolfiporia extensa. J Mushrooms 15: 54-56. https://doi.org/10.14480/JM.2017.15.1.54
  5. Jo WS, Yoo YB, Hong IP, Kim D. 2013. Changes of the cultivation methods of Poria cocos and its commercialization. J Mushrooms 11: 303-307. https://doi.org/10.14480/JM.2013.11.4.303
  6. Ka KH, Kim S, Park MJ, Jeong YS, Ryoo R, Jang Y, Choi JW, Kim SH. 2021. Development of new strains of Wolfiporia cocos for sclerotium formation by 2-way crossbreeding. Kor J Mycol 49: 405-412. https://doi.org/10.4489/KJM.20210038
  7. Kim YS. 1998. Effects of Poria cocos powder on wet noodle qualities. Appl Biol Chem 41: 539-544.
  8. Li S, Dong C. 2022. A new type of homokaryotic strain of Wolfiporia hoelen with indistinguishable phenotypes from the parent strains. Mycosystema 41: 1279-1292.
  9. Li S, Wang Q, Dong C. 2021. Distinguishing homokaryons and heterokaryons in medicinal polypore mushroom Wolfiporia cocos (Agaricomycetes) based on cultural and genetic characteristics. Front Microbiol 11: 596715. https://doi.org/10.3389/fmicb.2020.596715
  10. Li S, Wang Q, Dong C. 2022. Bipolar system of sexual incompatibility and heterothallic life cycle in the basidiomycetes Pachyma hoelen Fr. (Fuling). Mycologia 114: 63-75. https://doi.org/10.1080/00275514.2021.2006995
  11. Niculita-Hirzel H, Labbe J, Kohler A, Le Tacon F, Martin F, Sanders IR, Kues U. 2008. Gene organization of the mating type regions in the ectomycorrhizal fungus Laccaria bicolor reveals distinct evolution between the two mating type loci. New Phytol 180: 329-342. https://doi.org/10.1111/j.1469-8137.2008.02525.x
  12. Rocha de Brito M, Foulongne-Oriol M, Moinard M, Souza Dias E, Savoie JM, Callac P. 2016. Spore behaviors reveal a category of mating-competent infertile heterokaryons in the offspring of the medicinal fungus Agaricus subrufescens. Appl Microbiol Biotechnol 100: 781-796. https://doi.org/10.1007/s00253-015-7070-7
  13. Wang YZ, Zhang J, Zhao YL, Li T, Shen T, Li JQ, Li WY, Liu HG. 2013. Mycology, cultivation, traditional uses, phytochemistry and pharmacology of Wolfiporia cocos (Schwein.) Ryvarden et Gilb.: a review. J Ethnopharmacol 147: 265-276. https://doi.org/10.1016/j.jep.2013.03.027
  14. Zhao YY, Feng YL, Du X, Xi ZH, Cheng XL, Wei F. 2012. Diuretic activity of the ethanol and aqueous extracts of the surface layer of Poria cocos in rat. J Ethnopharmacol 114: 775-778.