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Enhancement of the germination efficiency of Gastrodia elata seeds using a new Mycena species

신규 애주름버섯 NIFOS101을 이용한 천마 종자의 발아효율 증진

  • Received : 2017.02.06
  • Accepted : 2017.03.15
  • Published : 2017.03.31

Abstract

This study investigated the in vitro symbiotic seed germination of the achlorophyllous orchid, Gastrodia elata, using a new Mycena species. A leaf-disc ($2{\times}2cm$) of Quercus accutissima was inoculated with either of the two fungal species, NIFOS101 (NCBI accession number KY449288) or KFRI1212 (HQ662845), following which it was placed on water agar medium, prior to sowing seeds of G. elata. After 20 days of symbiotic culture, NIFOS101 and KFRI1212 germinated 94% and 70% of G. elata seeds, respectively, although the initiation of protocorm development was induced much earlier by KFRI 1212 than by NIFOS101. Furthermore, the NIFOS101 mycelia grew much faster than KFRI 1212 at all temperatures tested. A phylogenetic analysis using the internal transcribed spacer (ITS) sequences showed that NIFOS101 belonged to a clade with M. purpureofusca, which completely differed from the clade with KFRI1212. This study not only identified a new fungal species, NIFOS101, which improved the rate of symbiotic seed germination up to 94% as compared to KFRI1212 (70%), but also revealed that G. elata required a broad taxonomic range of fungi for its symbiotic germination.

천마는 광합성 기능이 퇴화된 난과식물로 독립적인 생육이 불가능하다. 생장 단계에 따라 애주름버섯균(Mycena)과 뽕나무버섯균(Amillaria mellea)을 이용하여 유성증식법으로 생산한다. 본 연구에서는 신규 발아균주인 NIFOS101과 기존의 KFRI1212를 이용하여 천마종자의 발아효율과 원구경의 생장을 비교하였다. 두 균주를 낙엽미강배지에 접종한 후 천마 종자가 파종된 WA배지에 옮겨 배양하였다. 그 결과 NIFOS101과 KFRI1212를 이용한 발아율은 각각 94%와 70%이었으며, NIFOS101을 이용할 경우 KFRI1212에 비하여 원구경의 발달이 빠른 것으로 확인되었다. 또한 배양온도에 따른 균사의 생장력을 비교한 결과 NIFOS101이 KFRI1212 보다 균사의 생장이 빠른 것을 확인하였다. ITS 영역의 염기서열을 비교한 결과 애주름버섯속에서 NIFOS101은 붉은둘레애주름버섯과 유연관계가 높은 반면에 KFRI1212와는 유연관계가 낮은 것으로 나타났다. 본 연구에서는 천마의 신규 발아균으로 NIFOS101을 동정하고 천마의 기내 발아율 증진과 원구경의 생장 증진을 확인함으로서 향후 천마의 생산성을 높이는데 도움이 될 것으로 판단된다.

Keywords

References

  1. Bae KH, Ko MS, Choi SA, Lee HB, Kim NY, Song JM and Song GP (2012) In vitro germination of Gastrodia verrucosa Blume and Hetaeria sikokiana Tuyama treated by NaOCl. J Plant Biotechnol 39:163-168 https://doi.org/10.5010/JPB.2012.39.3.163
  2. Bak WC, Lee BH and Ka KH (2009) Characteristics of new shiitake strain "Sanlim No. 7" produced by di-mon hybridization method. Korean J Mycol 38(1):25-28 https://doi.org/10.4489/KJM.2010.38.1.025
  3. Bin H and Chen F (2004) Preparative isolation and purification of gastrodin from the chinese medicinal plant Gastrodia elata by high-speed counter current chromatography. J Chromatogr A 1052:229-232 https://doi.org/10.1016/j.chroma.2004.09.005
  4. Chang HM and But PH (1986) Pharmacology and applica-tion of chinese materia medica. World Scientific Publishing, Singapore, 185
  5. Han MH, Kim KD, Pyee JH, Choi SM and Park DS (2016) Observation of Scleropthora macrospora causing downy mildew from Zoysiagrass with leaf yellowing and excessive tillering. Weed Turf Sci 5:23-28 https://doi.org/10.5660/WTS.2016.5.1.23
  6. Hayashi J, Sekine T, Deguchi S, Lin Q, Horie S, Tsuchiya S, Yano S, Watanabe K and Ikegami F (2002) Phenolic compounds from Gastrodia rhizoma and relaxant effects of related compounds on isolated smooth muscle preparation. Phytochemistry 59:513-519 https://doi.org/10.1016/S0031-9422(02)00008-0
  7. Hong IP, Nam SH, Jung IY, Sung GB and Nam HW (2004) Studies on the conditions of seed germination of Gastrodia elata. Korean J Mycol 32:39-44 https://doi.org/10.4489/KJM.2004.32.1.039
  8. Huang ZL (1985) Recent developments in pharmacological study and clinical application of Gastrodia elata in china. Chung-His-I-Chieh-Ho-Tsa-Chih 5:251-254
  9. Kim HT, Kim JA and Park EJ (2012) Genetic diversity and metabolite analysis of Gastrodia elata by inter-simple sequence repeats (ISSR) markers. Korean J Medicinal Crop Sci 20:440-446 https://doi.org/10.7783/KJMCS.2012.20.6.440
  10. Kim HT and Park EJ (2013) Change of major functional components of Gastrodia elata Blume with cultivation conditions and harvest times. Korean J Medicinal Crop Sci 21:282-288 https://doi.org/10.7783/KJMCS.2013.21.4.282
  11. Kim HT, Kim ST, Lee WY and Park EJ (2013) Induction and growth of vegetative stems through In vitro culture of Gastrodia elata. Korean J Medicinal Crop Sci 21:142-147 https://doi.org/10.7783/KJMCS.2013.21.2.142
  12. Lee BY, Choi HS and Hwang JB (2002) Analysis of food components of Gastrodia rhizoma and changes in several characteristics at the various drying conditions. Korean J Food Sci Technol 34:37-42
  13. Lee JY (1982) The propagation of Gastrodia elata by using rhizomorphs of Armillariella mella. Seoul Women's University. Rural Development Res Service 7:39-45
  14. Park EJ, Lee WY and Ahn JK (2012) In vitro propagation of myco-heterotrophic Gastrodia elata. Hort Environ Biotechnol 53:415-420 https://doi.org/10.1007/s13580-012-0046-y
  15. Park EJ and Lee WY (2013) In vitro symbiotic germination of myco-heterotrophic Gastrodia elata by Mycena species. Plant Biotechnol Rep 7:185-191 https://doi.org/10.1007/s11816-012-0248-x
  16. Smith SE and Read DJ (1997) Mycorrhizal symbiosis (2nd ed). Academic Press. San Diego
  17. White TJ, Bruns T, Lee S and Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ(ed.). PCR Protocols. A Guide to Methods and Applications. Academic Press. NY. USA. pp.315-322