DOI QR코드

DOI QR Code

원형질체융합 기법을 이용한 산느타리 계통육성

Breeding of new variety Pleurotus pulmonarius using protoplast fusion technique

  • 권희민 (국립농업과학원 농식품자원부 발효가공식품과) ;
  • 이윤혜 (국립농업과학원 농업환경부 토양비료과) ;
  • 김정한 (경기도농업기술원 친환경미생물연구소) ;
  • 백일선 (경기도농업기술원 친환경미생물연구소) ;
  • 강희완 (한경대학교 생명공학부 원예생명공학전공) ;
  • 최종인 (경기도농업기술원 환경농업연구과)
  • Gwon, Hee-Min (Fermented Processing Food Science Division, Department of Agrofood Resource, NIAS, RDA) ;
  • Lee, Yun-Hae (Soil and Fertilizer Division, National Institute of Agricultural Sciences) ;
  • Kim, Jeong-Han (Eco-friendly Environment & Microorganism Research Institute, Gyenggi-Do Agricultural Research & Extension services) ;
  • Baek, Il-Sun (Eco-friendly Environment & Microorganism Research Institute, Gyenggi-Do Agricultural Research & Extension services) ;
  • Kang, Hee-Wan (Department of Holticultural Biotechnology, Division of Biotechnology, Hankyong National University) ;
  • Choi, Jong-In (Department of environmental agriculture research, Gyenggi-Do Agricultural Research & Extension services)
  • 투고 : 2021.09.01
  • 심사 : 2021.09.25
  • 발행 : 2021.09.30

초록

원형질체 융합 기술은 종·속간 유전적 한계를 넘어 육종과 그 소재로 활용하고데 목적이 있다. 본 연구에서는 '흑타리'(P. ostreatus)와 '호산'(P. pulmonarius)의 단핵균사를 이용하여 원형질체를 나출하고 나출된 원형질체를 융합하여 종간 교배 계통을 육성하였다. 육성계통의 균사생장속도는 '호산', '흑타리', PF160313, PF160306 계통 순으로 빠른 편이었다. 균사 밀도는 PF160306 계통이 가장 높았고, 나머지는 중간 수준의 밀도를 나타내었다. 원형질체 융합계통인 PF160306과 PF160313 계통은 '흑타리' 품종보다 배양 기간이 10일, '호산' 품종보다 2일 단축되었다. 자실체 생장 기간은 '흑타리'와 '호산에 비하여 각각 3일, 1일 단축되었다. PF160306 계통의 생산량은 135.9 g/병으로 '호산'에 비하여 높았으나 통계적으로 유의차가 없었다. 자실체 발생기간은 15℃에서 9일, 25℃에서 4.5일로 온도가 높아짐에 빨라졌다. 자실체의 갓색은 21℃ 노란색이 가장 선명하게 발현되었다. URP primer 7을 사용하여 PCR 밴드 패턴을 비교하였을 때, 전체적으로 '호산' 품종과 유사하였다. DPPH radical 소거능과 폴리페놀 함량에 있어 '순정'은 각각 62.5%, 43.5 mg/mL였으며, PF160313 계통은 각각 65.7%, 49.9 mg/mL를 나타내어 계통간 유의차가 있었다. ACE 활성은 '순정' 74%, PF160313 계통 75%로 유사한 수준이었다.

In this study, monokaryons of "Heukari" (Pleurotus ostreatus) and "Hosan" (Pleurotus pulmonarius) were separated to remove the cell wall, and a cross-species protoplast fusion was developed through chemical treatment with polyethylene glycol. The protoplast-fused PF160306 and PF160313 strains have a culture period of 10 and 2 days shorter than that of the "Heuktari" and "Hosan" cultivars, respectively. Furthermore, the growth of the strains was faster than that of the existing cultivars. The yield was 135.9 g per bottle, which was approximately 8% higher than that of the commercially available "Hosan" cultivar; however, it was not statistically significant. A growth survey was conducted after treatment at five temperatures (15, 18, 21, 23, and 25℃). The growth of the strains accelerated with the increase in temperature. However, at 21℃, the yellow color of pileus was the brightest. Band pattern, assessed using URP Primer 7, was similar to that of the "Hosan" cultivar. The DPPH radical scavenging capacity and polyphenol content were 62.5% and 43.5 mg/mL, respectively, for "Sunjung" and 65.7% and 49.9 mg/mL, respectively, for PF160313. Furthermore, the antihypertensive activities of the "Sunjung" cultivar and PF160313 were similarly high at 74% and 75%, respectively. In conclusion, cross-species hybridization via the protoplast fusion technique can be used for obtaining primary data for mushroom breeding to develop new varieties. In addition, the protoplast fusion technique might aid in expanding the market for yellow mushrooms.

키워드

과제정보

본 결과물은 농림축산식품부의 재원으로 농림식품기술기획평가원의 Golden Seed 프로젝트 사업의 지원을 받아 연구되었으며(213007-05-5-SB120) 이에 감사드립니다.

참고문헌

  1. Anne J and Peberdy JF. 1976. Induced fusion of fungal protoplasts following treatment with polyethylene glycol. J Gen Microbiol 92: 413-417. https://doi.org/10.1099/00221287-92-2-413
  2. Cho JH, Park HS, Han JG, Lee GH, Sung GH and Jhune CS. 2014. Comparative analysis of anti-oxidant effects and polyphenol contents of the fruiting bodies in oyster mushrooms. J Mushrooms 12: 311-315. https://doi.org/10.14480/JM.2014.12.4.311
  3. Cho KS and Park TH. 2014. Potato breeding via protoplast fusion. J Plant Biotechnol 41:65-72. https://doi.org/10.5010/JPB.2014.41.2.65
  4. Dhitaphichit P and Pornsuriya C. 2005. Protoplast fusion between Pleurotus ostreatus and P. djamor. Songklanakarin. J Sci Technol 27: 975-982.
  5. Djajanegara I and Masduki A. 2010. Protoplast fusion between white and brown oyster mushrooms. Indones J Agric Sci 11: 16-23. https://doi.org/10.21082/ijas.v11n1.2010.p16-23
  6. Jang IJ, Chung KC and Chang HY. 2005. Excellent strain selection and optimal mycelial growth condition of Pleurotus cornucopiae. The Korean Society of Mushroom Science 3: 40-44.
  7. Jeon D H, Lee YH, Choi JI, Gwon HM and Chi JH. 2018. Characteristics of a new Grifola frondosa Cultivar "Daebak" with stable pinheading and high yield. J Mushrooms 16: 203-207.
  8. Jhune CS, Kong WS, Yoo YB, Jang KY, Paik SB and Chun SC. 2006. Initiation and growth of fruitbody of oyster mushroom as affected by cultivation temperature. J Mushrooms 4: 33-38.
  9. Kang HW. 2012. Genetic diversity analysis of fungal species by universal rice primer(URP)-PCR. Kor J Mycol 40: 78-85. https://doi.org/10.4489/KJM.2012.40.2.78
  10. Kiguchi T and Yanagi SO. 1985. Intraspecific heterokaryon and fruit body formation in Coprinus macrorhizus by protoplast fusion of auxotrophic mutants. Appl Microbiol Biotechnol 22: 121-127. https://doi.org/10.1007/BF00250031
  11. Kim EJ, Shin PG, Jang GY, Kong WS, Han YS and Yoo YB. 2014. Nuclear DNA inheritance of intra-specific somatic hybricds by di-mono cross in Pleurotus ostreatus based on URP-PCR ananlysis. J Mushrooms 12: 96-106. https://doi.org/10.14480/JM.2014.12.2.96
  12. Lee HJ, Do JR, Chung MY and Kim HK. 2014. Antioxidant activities of Pleurotus cornucopiae extracts by extraction conditions. J Korean Soc Food Sci Nutr 43: 836-841. https://doi.org/10.3746/jkfn.2014.43.6.836
  13. Mallick P and Sikdar SR. 2014. Production and molecular characterization of somatic hybrids between Pleurotus florida and Lentinula edodes. World J Microbiol Biotechnol 30:2283-2293. https://doi.org/10.1007/s11274-014-1652-x
  14. Oh MJ, Kim EJ, Jung JH, Shin PG, Kim ES, Oh YL, Jang KY, Kong WS and Yoo YB. 2015. Characterization of a new commercial strain 'Mongdol' by intra-specific hyphal anastomosis in Pleurotus ostratus. J Mushrooms 13: 212-216. https://doi.org/10.14480/JM.2015.13.3.212
  15. Park JH, Kang MS, Kim HI, Chung BH, Lee KH and Moon WK. 2003. Study on immuno-stimulating activity of β-glucan isolated from the cell wall of yeast mutant Saccharomyces cerevisiae IS2. Korean J Food Sci Technol 35: 488-492.
  16. Peberdy JF. 1995. Fungal Protoplasts. Genetics and Biotechnology. K. Ulrich(Ed), p.49-60.
  17. Swain T and Hillis WE. 1959. Phenolic constituents of Prunus domestica I quantitative analysis of phenolic constituents. J Sci Food Agric 10: 63-68. https://doi.org/10.1002/jsfa.2740100110
  18. Toyomatsu T and Mori KI. 1987. Intra- and inter-specific protoplast fusion between some Pleurotus species. Agric Biol Chem 51: 935-937. https://doi.org/10.1271/bbb1961.51.935
  19. Um SN, Jin GE, Park KW, Yu YB and Park KM. 2010. Physiological Activity and Nutritional Composition of Pleurotus Species. Korean J Food Sci Technol 42: 90-96.
  20. Williams WB, Cuvelier ME and Berset C. 1994. Use of a free radical method to evaluate antioxidant activity. Lebensm-Wiss u-Technol 28: 25-30.
  21. Yoo YB, You CH and Cha DY. 1993. Strain improvement of the genus Pleurotus by protoplast fusion. Kor J Mycol 21: 200-211.
  22. Yoo YB, Kong WS, Jang KY, Kim IY, Oh SJ and Jhune CS. 2006. Characterization of a new commercial strain 'Gumbit'. J Mushrooms 4: 83-87.
  23. Yoo YB, Oh MJ, Oh YL, Shin PG, Jang GY and Kong WS. 2016. Development trend of the mushroom industry. J Mushrooms 14: 142-154. https://doi.org/10.14480/JM.2016.14.4.142