An optimized isolation protocol yields high-quality RNA from cassava tissues (Manihot esculenta Crantz)

被引:18
作者
Behnam, Babak [1 ]
Bohorquez-Chaux, Adriana [2 ]
Fernando Castaneda-Mendez, Oscar [2 ]
Tsuji, Hiroyuki [1 ]
Ishitani, Manabu [2 ]
Becerra Lopez-Lavalle, Luis Augusto [2 ]
机构
[1] Yokohama City Univ, Kihara Inst Biol Res, Yokohama, Kanagawa, Japan
[2] Int Ctr Trop Agr CIAT, Recta Cali Palmira Km 17, Valle Del Cauca, Colombia
来源
FEBS OPEN BIO | 2019年 / 9卷 / 04期
基金
比尔及梅琳达.盖茨基金会;
关键词
cassava; qRT-PCR; RNA isolation; RNA sequencing; EFFICIENT PROTOCOL; TRANSCRIPTOME ANALYSIS; FUNCTIONAL RNA; EXTRACTION; DROUGHT; RICH;
D O I
10.1002/2211-5463.12561
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We developed and modified a precise, rapid, and reproducible protocol isolating high-quality RNA from tissues of multiple varieties of cassava plants (Manihot esculenta Crantz). The resulting method is suitable for use in mini, midi, and maxi preparations and rapidly achieves high total RNA yields (170-600gg(-1)) using low-cost chemicals and consumables and with minimal contamination from polysaccharides, polyphenols, proteins, and other secondary metabolites. In particular, A(260):A(280) ratios were >2.0 for RNA from various tissues, and all of the present RNA samples yielded ribosomal integrity number values of greater than six. The resulting high purity and quality of isolated RNA will facilitate downstream applications (quantitative reverse transcriptase-polymerase chain reaction or RNA sequencing) in cassava molecular breeding.
引用
收藏
页码:814 / 825
页数:12
相关论文
共 32 条
  • [1] Overexpression of Arabidopsis FLOWERING LOCUS T (FT) gene improves floral development in cassava (Manihot esculenta, Crantz)
    Adeyemo, O. Sarah
    Chavarriaga, Paul
    Tohme, Joe
    Fregene, Martin
    Davis, Seth J.
    Setter, Tim L.
    [J]. PLOS ONE, 2017, 12 (07):
  • [2] Amuge T, 2017, SCI REP, V6, P32006
  • [3] Transcriptome profiling of low temperature-treated cassava apical shoots showed dynamic responses of tropical plant to cold stress
    An, Dong
    Yang, Jun
    Zhang, Peng
    [J]. BMC GENOMICS, 2012, 13
  • [4] Inversion induced Manihot esculenta stem tubers express key tuberization genes; Mec1, RZF, SuSy1 and PIN2
    Bowrin, Valerie
    Sutton, Fedora
    [J]. PLANT SIGNALING & BEHAVIOR, 2016, 11 (01)
  • [5] A simple hydroponic hardening system and the effect of nitrogen source on the acclimation of in vitro cassava (Manihot esculenta Crantz)
    Castaneda-Mendez, Oscar
    Ogawa, Satoshi
    Medina, Adriana
    Chavarriaga, Paul
    Gomez Selvaraj, Michael
    [J]. IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-PLANT, 2017, 53 (02) : 75 - 85
  • [6] Chang S. J., 1993, Plant Molecular Biology Reporter, V11, P113, DOI 10.1007/BF02670468
  • [7] Chaudhary S, 2016, J ADV RES BIOTECH, V1, P7
  • [8] CHOMCZYNSKI P, 1987, ANAL BIOCHEM, V162, P156, DOI 10.1016/0003-2697(87)90021-2
  • [9] Chun J, 2008, PROG BIOCHEM BIOPHYS, V35, P591
  • [10] A simple and efficient protocol for isolation of high quality functional RNA from different tissues of turmeric (Curcuma longa L.)
    Deepa, K.
    Sheeja, T. E.
    Santhi, R.
    Sasikumar, B.
    Cyriac, Anu
    Deepesh, P. V.
    Prasath, D.
    [J]. PHYSIOLOGY AND MOLECULAR BIOLOGY OF PLANTS, 2014, 20 (02) : 263 - 271