Plant Physiology

Choosing of Methods for Total RNA Extraction from Malus sieversii Fruits in Xinjiang

  • XIA Chun-lan ,
  • WEN Zhi-bin ,
  • ZHANG Yu-fang
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  • 1. Key Laboratory of Biogeography and Biological Resources under Chinese Academy of Sciences,Xinjiang institute of Ecology and Geography,Chinese Academy of Sciences,Urumqi 830011,Xinjiang,China;
    2. University of Chinese Academy of Sciences,Beijing 101408,China;
    3. College of Life Science,Shihezi University,Shihezi 832000,Xinjiang,China

Received date: 2018-04-24

  Revised date: 2018-08-30

  Online published: 2025-10-18

Abstract

Malus sieversii fruits are rich in polysaccharides,polyphenols,soluble pectin and other secondary metabolites. Therefore,it is relatively difficult to extract the total RNA from M.sieversii fruits. In order to choose an effective extraction method,five RNA extraction methods including the TaKaRa Kit,Trans Kit,improved CTAB method by Tian Wei,modified CTAB method I and CTAB method Ⅱwere compared using the fruits at five different developmental stages. Results showed that neither kits method was able to be used to extract the RNA from M.sieversii fruits. The quality of total RNA extracted with three modified CTAB methods was good,with two distinct electrophoresis bands of 28S and 18S rRNA,the OD260/OD280 values ranged from 1.8 to 2.2,and the OD260/OD230 value was higher than 2.0. But only the modified CTAB method I was an effective method to yield high quantity of total RNA. The average extraction values of fruits at five different developmental stages were ranged from 116.79 μg·g-1 to 474.76 μg·g-1. For the modified CTAB method I,the impurity in the RNA was effectively removed by extending LiCl precipitation time and placing extraction under 4 ℃ for overnight aging (not longer than 16 hours)instead of -20 ℃ for 15 min. Although this method took 2 days,the method was easy to operate,and the extraction cost was low. Therefore,the modified CTAB method I is an effective method for total RNA extraction from M.sieversii fruits (especially for the ripe fruits),which could be used in further molecular biology research.

Cite this article

XIA Chun-lan , WEN Zhi-bin , ZHANG Yu-fang . Choosing of Methods for Total RNA Extraction from Malus sieversii Fruits in Xinjiang[J]. Arid Zone Research, 2019 , 36(2) : 437 -443 . DOI: 10.13866/j.azr.2019.02.20

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