Mycobiology

The Korean Society of Mycology (한국균학회)
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An Improved Total RNA Extraction Method for White Jelly Mushroom Tremella fuciformis Rich in Polysaccharides

  • Zhu, Hanyu (College of Food Science and Technology, Huazhong Agricultural University) ;
  • Sun, Xueyan (College of Food Science and Technology, Huazhong Agricultural University) ;
  • Liu, Dongmei (College of Food Science and Technology, Huazhong Agricultural University) ;
  • Zheng, Liesheng (College of Plant Science and Technology, Huazhong Agricultural University) ;
  • Chen, Liguo (College of Plant Science and Technology, Huazhong Agricultural University) ;
  • Ma, Aimin (College of Food Science and Technology, Huazhong Agricultural University)
  • Received : 2017.09.11
  • Accepted : 2017.11.30
  • Published : 2017.12.01
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Abstract

An improved method for extracting high quality and quantity RNA from a jelly mushroom and a dimorphic fungus-Tremella fuciformis which is especially rich in polysaccharides, is described. RNA was extracted from T. fuciformis mycelium M1332 and its parental monokaryotic yeast-like cells Y13 and Y32. The A260/280 and A260/230 ratios were both approximately 2, and the RNA integrity number was larger than 8.9. The yields of RNA were between 108 and $213{\mu}g/g$ fresh wt. Downstream molecular applications including reverse transcriptional PCR and quantitative real-time PCR were also performed. This protocol is reliable and may be widely applicable for total RNA extraction from other jelly mushrooms or filamentous fungi rich in polysaccharides.

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References

  1. Zhu H, Yuan Y, Liu J, Zheng L, Chen L, Ma A. Comparing the sugar profiles and primary structures of alkali-extracted water-soluble polysaccharides in cell wall between the yeast and mycelial phases from Tremella fuciformis. J Microbiol 2016;54:381-6. https://doi.org/10.1007/s12275-016-5533-x
  2. Nickerson KW, Atkin AL. Deciphering fungal dimorphism: Farnesol's unanswered questions. Mol Microbiol 2017;103: 567-75. https://doi.org/10.1111/mmi.13601
  3. Hou L, Chen Y, Ma C, Liu J, Chen L, Ma A. Effects of environmental factors on dimorphic transition of the jelly mushroom Tremella fuciformis. Cryptogam Mycol 2011;32: 421-8. https://doi.org/10.7872/crym.v32.iss4.2011.421
  4. Li Y, Wang W, Du X, Yuan Q. An improved RNA isolation method for filamentous fungus Blakeslea trispora rich in polysaccharides. Appl Biochem Biotechnol 2010;160:322-7. https://doi.org/10.1007/s12010-008-8289-x
  5. Muoki RC, Paul A, Kumari A, Singh K, Kumar S. An improved protocol for the isolation of RNA from roots of tea (Camellia sinensis (L.) O. Kuntze). Mol Biotechnol 2012;52: 82-8. https://doi.org/10.1007/s12033-011-9476-5
  6. Li JH, Tang CH, Song CY, Chen MJ, Feng ZY, Pan YJ. A simple, rapid and effective method for total RNA extraction from Lentinula edodes. Biotechnol Lett 2006;28:1193-7. https://doi.org/10.1007/s10529-006-9074-y
  7. Kumar A, Singh K. Isolation of high quality RNA from Phyllanthus emblica and its evaluation by downstream applications. Mol Biotechnol 2012;52:269-75. https://doi.org/10.1007/s12033-011-9492-5
  8. Ghawana S, Paul A, Kumar H, Kumar A, Singh H, Bhardwaj PK, Rani A, Singh RS, Raizada J, Singh K, et al. An RNA isolation system for plant tissues rich in secondary metabolites. BMC Res Notes 2011;4:85. https://doi.org/10.1186/1756-0500-4-85
  9. Deng LH, Luo MW, Zhang CF, Zeng HC. Extraction of highquality RNA from rubber tree leaves. Biosci Biotechnol Biochem 2012;76:1394-6. https://doi.org/10.1271/bbb.120014
  10. Rajakani R, Narnoliya L, Sangwan NS, Sangwan RS, Gupta V. Activated charcoal-mediated RNA extraction method for Azadirachta indica and plants highly rich in polyphenolics, polysaccharides and other complex secondary compounds. BMC Res Notes 2013;6:125. https://doi.org/10.1186/1756-0500-6-125
  11. Fleige S, Pfaffl MW. RNA integrity and the effect on the realtime qRT-PCR performance. Mol Aspects Med 2006;27:126-39 https://doi.org/10.1016/j.mam.2005.12.003
  12. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the $2^{-{\Delta}{\Delta}Ct}$ method. Methods 2001;25:402-8. https://doi.org/10.1006/meth.2001.1262