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An Improved Total RNA Extraction Method for White Jelly Mushroom Tremella fuciformis Rich in Polysaccharides 원문보기

Mycobiology, v.45 no.4, 2017년, pp.434 - 437  

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)

Abstract AI-Helper 아이콘AI-Helper

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. ...

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제안 방법

  • 8% (w/v) agarose gel. The purity and quantity of RNA was tested by evaluating the ratio of A260/280 and A260/230 using a NanoDrop 2000 spectrophotometer (Thermo Scientific, Waltham, MA, USA). RNA integrity number (RIN) was examined using an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA).

대상 데이터

  • A dikaryotic strain of T. fuciformis M1332 (mycelial form) and two parental monokaryotic strains Y13 and Y32 (yeast form), were obtained from the Culture Collection of State Key Laboratory of Agricultural Microbiology which is the part of China Center for Type Culture Collection (CCTCC). The strains were incubated at 25℃ for 7 days to harvest yeast-like cells and 20 days to obtain mycelium.

이론/모형

  • Each qRT-PCR reaction was carried out with independent triplicate trials and the expression ratios were calculated according to the 2−ΔΔCt method [12].
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참고문헌 (12)

  1. 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 54 381 386 27095457 

  2. 2 Nickerson KW Atkin AL Deciphering fungal dimorphism: Farnesol's unanswered questions Mol Microbiol 2017 103 567 575 27987234 

  3. 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 32 421 428 

  4. 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 160 322 327 18561031 

  5. 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 88 22144070 

  6. 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 1197 16799758 

  7. 7 Kumar A Singh K Isolation of high quality RNA from Phyllanthus emblica and its evaluation by downstream applications Mol Biotechnol 2012 52 269 275 22209897 

  8. 8 Ghawana S Paul A Kumar H Kumar A Singh H Bhardwaj PK Rani A Singh RS Raizada J Singh K An RNA isolation system for plant tissues rich in secondary metabolites BMC Res Notes 2011 4 85 21443767 

  9. 9 Deng LH Luo MW Zhang CF Zeng HC Extraction of high-quality RNA from rubber tree leaves Biosci Biotechnol Biochem 2012 76 1394 1396 22785464 

  10. 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 23537338 

  11. 11 Fleige S Pfaffl MW RNA integrity and the effect on the real-time qRT-PCR performance Mol Aspects Med 2006 27 126 139 16469371 

  12. 12 Livak KJ Schmittgen TD Analysis of relative gene expression data using real-time quantitative PCR and the 2 −ΔΔCt method Methods 2001 25 402 408 11846609 

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