Research article Topical Sections

How to store plant tissues in the absence of liquid nitrogen? Ethanol preserves the RNA integrity of Cannabis sativa stem tissues

  • Received: 07 July 2016 Accepted: 26 September 2016 Published: 28 September 2016
  • The preservation of intact RNA is a limiting step when gene expression profiling is performed using field-collected plant material. The use of liquid nitrogen ensures the optimal preservation of RNA, however it is not always practical, especially if the plant material has to be sampled in remote locations. Ethanol is known to preserve DNA in plant tissues even after a long storage period and here its suitability to preserve the RNA of textile hemp cortical tissues was tested. Hemp (Cannabis sativa L.) is an economically important fibre crop because it supplies cellulosic bast fibres used in different industrial sectors. In this study we demonstrate the suitability of ethanol for RNA preservation by analyzing tissues stored at 4 °C for 1, 2, 4 and 8 days. We show that in all the cases the extracted RNA is intact. We finally analyze hemp stem tissues stored in ethanol for 1 month and demonstrate the preservation of the tissue structure, particularly of bast fibres.

    Citation: Lauralie Mangeot-Peter, Sylvain Legay, Jean-Francois Hausman, Gea Guerriero. How to store plant tissues in the absence of liquid nitrogen? Ethanol preserves the RNA integrity of Cannabis sativa stem tissues[J]. AIMS Molecular Science, 2016, 3(4): 560-566. doi: 10.3934/molsci.2016.4.560

    Related Papers:

  • The preservation of intact RNA is a limiting step when gene expression profiling is performed using field-collected plant material. The use of liquid nitrogen ensures the optimal preservation of RNA, however it is not always practical, especially if the plant material has to be sampled in remote locations. Ethanol is known to preserve DNA in plant tissues even after a long storage period and here its suitability to preserve the RNA of textile hemp cortical tissues was tested. Hemp (Cannabis sativa L.) is an economically important fibre crop because it supplies cellulosic bast fibres used in different industrial sectors. In this study we demonstrate the suitability of ethanol for RNA preservation by analyzing tissues stored at 4 °C for 1, 2, 4 and 8 days. We show that in all the cases the extracted RNA is intact. We finally analyze hemp stem tissues stored in ethanol for 1 month and demonstrate the preservation of the tissue structure, particularly of bast fibres.


    加载中
    [1] Guerriero G, Hausman J-F, Strauss J, et al (2016) Lignocellulosic biomass: Biosynthesis, degradation, and industrial utilization. Eng Life Sci 16: 1-16. doi: 10.1002/elsc.201400196
    [2] De Pauw MA, Vidmar JJ, Collins J, et al. (2007) Microarray analysis of bast fibre producing tissue of Cannabis sativa identifies transcripts associated with conserved and specialised processes of secondary wall development. Func Plant Biol 34: 737-749. doi: 10.1071/FP07014
    [3] Van den Broeck HC, Maliepaard C, Ebskam MJM, et al. (2008) Differential expression of genes involved in C1 metabolism and lignin biosynthesis in wooden core and bast tissues of fibre hemp (Cannabis sativa L.). Plant Sci 2: 205-220.
    [4] Bainard LD, Klironomos JN, Hart MM (2010) Differential effect of sample preservation methods on plant and arbuscular mycorrhizal fungal DNA. J Microbiol Methods 82: 124-130. doi: 10.1016/j.mimet.2010.05.001
    [5] Bressan EA, Rossi ML, Gerald LT, et al. (2014) Extraction of high-quality DNA from ethanol-preserved tropical plant tissues. BMC Res Notes 7: 268. doi: 10.1186/1756-0500-7-268
    [6] Cooper A (1994) DNA from museum specimens, In: Hermann B. and Hummel S., eds. Ancient DNA, New York: Springer-Verlag, 49-165.
    [7] Linke B, Schröder K, Arter J, et al. (2010) Extraction of nucleic acids from yeast cells and plant tissues using ethanol as medium for sample preservation and cell disruption. Biotechniques 49: 655-657. doi: 10.2144/000113476
    [8] Lou JJ, Mirsadraei L2, Sanchez DE, et al. (2014) A review of room temperature storage of biospecimen tissue and nucleic acids for anatomic pathology laboratories and biorepositories. Clin Biochem 47: 267-273. doi: 10.1016/j.clinbiochem.2013.12.011
    [9] Tuorto SJ, Brown CM, Bidle KD, et al. (2015) BioDry: an inexpensive, low-power method to preserve aquatic microbial biomass at room temperature. PLoS One 10: e0144686. doi: 10.1371/journal.pone.0144686
    [10] Alexandersson E, Jacobson D, Vivier MA, et al. (2014) Field-omics-understanding large-scale molecular data from field crops. Front Plant Sci 5: 286.
    [11] Guerriero G, Mangeot-Peter L, Hausman J-F, Legay S (2016) Extraction of high quality RNA from Cannabis sativa bast fibres: a vademecum for molecular biologists. Fibers 4: 23.
    [12] Neutelings G (2011) Lignin variability in plant cell walls: contribution of new models. Plant Sci 181: 379-386. doi: 10.1016/j.plantsci.2011.06.012
    [13] Guerriero G, Sergeant K, Hausman JF (2013) Integrated -omics: a powerful approach to understanding the heterogeneous lignification of fibre crops. Int J Mol Sci 14: 10958-10978. doi: 10.3390/ijms140610958
    [14] Maier TS, Kuhn J, Müller C (2010) Proposal for field sampling of plants and processing in the lab for environmental metabolic fingerprinting. Plant Methods 6: 6. doi: 10.1186/1746-4811-6-6
    [15] Andre CM, Hausman JF, Guerriero G (2016) Cannabis sativa: the plant of the thousand and one molecules. Front Plant Sci 7: 19.
    [16] Guerriero G, Sergeant K, Hausman JF (2014) Wood biosynthesis and typologies: a molecular rhapsody. Tree Physiol 34: 839-855. doi: 10.1093/treephys/tpu031
    [17] Parrotta L, Guerriero G, Sergeant K, et al (2015) Target or barrier? The cell wall of early- and later-diverging plants vs cadmium toxicity: differences in the response mechanisms. Front Plant Sci 6: 133.
    [18] Natsume S, Takagi H, Shiraishi A, et al (2015) The draft genome of hop (Humulus lupulus), an essence for brewing. Plant Cell Physiol 56: 428-441 doi: 10.1093/pcp/pcu169
  • Reader Comments
  • © 2016 the Author(s), licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0)
通讯作者: 陈斌, bchen63@163.com
  • 1. 

    沈阳化工大学材料科学与工程学院 沈阳 110142

  1. 本站搜索
  2. 百度学术搜索
  3. 万方数据库搜索
  4. CNKI搜索

Metrics

Article views(5694) PDF downloads(1079) Cited by(0)

Article outline

Figures and Tables

Figures(3)  /  Tables(1)

/

DownLoad:  Full-Size Img  PowerPoint
Return
Return

Catalog