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A Modified Protocol for Total Rna Isolation from Different Oil Palm (Elaeis guineensis) Tissues using Cetyltrimethylammonium Bromide


Affiliations
1 Hainan Key Laaboratory of Tropical Oil Crops Biology/Coconut Research Institute, Chinese Academy of Tropical Agricultural Science, Wenchang, Hainan 571339, China
2 Department of Environmental Sciences, Lahore College for Women University Lahore, Lahore 54600, Pakistan
3 Institute of Horticultural Sciences, University of Agriculture, Faisalabad, Pakistan
 

Extraction of high-quality RNA from oil palm tissues is challenging due to the presence of polysaccharides, polyphenols and other complexes that co-precipitate with RNA. Therefore, isolation of high-quality RNA from oil palm is challenging due to the presence of varying amounts of these constituents in diverse tissues. This communication describes a modified RNA extraction protocol based on the cetyltrimethylammonium bromide (CTAB) method which is useful for extracting high-quality RNA from different oil palm tissues. Total RNA isolation using a modified CTAB protocol was compared with two different methods, a conventional TRIzol method and the method for RNA isolation from palms (MRIP). Both methods were useful for isolating RNA from leaf tissues; however, they were not effective in isolating RNA from other tissues. The current protocol based on a modified CTAB method was efficient in isolating high-quality total RNA from oil palm fruit tissues, including mesocarp and endosperm, stem, ischolar_main and flower tissues. This modified CTAB-based protocol gave approximately 20-30 μg of total RNA from 150 mg of tissue within 5-6 h.

Keywords

CTAB Method, Gene Expression, Oil Palm, RNA Extraction.
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  • Alain, R. and Larkins, B. A., Biotechnology in Agriculture and Forestry, Springer-Verlag, Heidelberg, Germany, 2007, vol. 63, pp. 59-80.

  • Nair, P., Oil palm (Elaeis guineensis Jacquin). In The Agronomy and Economy of Important Tree Crops of the Developing World, Elsevier Inc., London, UK, 2010, pp. 209-236.

  • Singh, R. P., Hakimi Ibrahim, M. Norizan Esa and Iliyana, M. S., Composting of waste from palm oil mill: a sustainable waste management practice. Rev. Environ. Sci. Biotechnol., 2010, 9, 331- 344.

  • Sambanthamurthi, R., Singh, R., Kadir, A. P. G., Abdullah, M. O. and Kushairi, A., Opportunities for the oil palm via breeding and biotechnology. In Breeding Plantation Tree Crops: Tropical Species, Springer, 2009, pp. 377-421.

  • Baud, S., Mendoza, M. S., To, A., Harscoët, E., Lepiniec, L. and Dubreucq, B., WRINKLED1 specifies the regulatory action of leafy cotyledon2 towards fatty acid metabolism during seed maturation in arabidopsis. Plant J., 2007, 50, 825-838.

  • Maeo, K. et al., An ap2-type transcription factor, wrinkled1, of arabidopsis thaliana binds to the aw-box sequence conserved among proximal upstream regions of genes involved in fatty acid synthesis. Plant J., 2009, 60, 476-487.

  • Helen, H. T., Pelletier, C. and Beardmore, T., Total RNA isolation from Picea mariana dry seed. Plant Mol. Biol. Rep., 2004, 22, 93a-93e.

  • Wang, X., Tian, W. and Li, Y., Development of an efficient protocol of rna isolation from recalcitrant tree tissues. Mol. Biotechnol., 2008, 38, 57-64.

  • Salzman, R. A., Fujita, T., Zhu-Salzman, K., Hasegawa, P. M. and Bressan, R. A., An improved RNA isolation method for plant tissues containing high levels of phenolic compounds or carbohydrates.

  • Plant Mol. Biol. Rep., 1999, 17, 11-17.

  • Li, J.-H., Tang, C.-H., Song, C.-Y., Chen, M.-J., Feng, Z.-Y. and Pan, Y.-J., A simple, rapid and effective method for total RNA extraction from Lentinula edodes. Biotechnol. Lett., 2006, 28, 1193-1197.

  • Chun Jun, Z. Y.-F., Sheng-Hua, W. and Fang, C., A RNA isolation method suitable for a wide range of materials. Prog. Biochem. Biophys., 2008, 35, 591-597.

  • Thanh, T., Omar, H., Abdullah, M. P., Chi, V. T. Q., Noroozi, M., Ky, H. and Napis, S., Rapid and effective method of RNA isolation from green microalga Ankistrodesmus convolutus. Mol. Biotechnol., 2009, 43, 148-153.

  • Takahashi, H. et al., A method for obtaining high quality RNA from paraffin sections of plant tissues by laser microdissection. J. Plant Res., 2010, 123, 807-813.

  • Tattersall, E. A., Ergul, A., AlKayal, F., DeLuc, L., Cushman, J. C. and Cramer, G. R., Comparison of methods for isolating high-quality RNA from leaves of grapevine. Am. J. Enol. Viticult., 2005, 56, 400-406.

  • Hou, P., Xie, Z., Zhang, L., Song, Z., Mi, J., He, Y. and Li, Y., Comparison of three different methods for total RNA extraction from Fritillaria unibracteata, a rare Chinese medicinal plant. J. Med. Plants Res., 2011, 5, 2835-2839.

  • Kiefer, E., Heller, W. and Ernst, D., A simple and efficient protocol for isolation of functional RNA from plant tissues rich in secondary metabolites. Plant Mol. Biol. Rep., 2000, 18, 33-39.

  • Bourgisa, F., Kilaruc, A., Caod, X., Ngando-Ebonguee, G.-F., Driraf, N., Ohlrogged, J. B. and Arondela, V., Comparative transcriptome and metabolite analysis of oil palm and date palm mesocarp that differ dramatically in carbon partitioning. Metabolism, 2011, 156, 564-584.

  • Tranbarger, T. J. et al., Regulatory mechanisms underlying oil palm fruit mesocarp maturation, ripening, and functional specialization in lipid and carotenoid metabolism. Plant Physiol., 2011, 156, 564-584.

  • Xiao, Y. et al., Efficient isolation of high quality RNA from tropical palms for RNA-Seq analysis. Plant Omics, 2012, 5, 584.

  • Bernardo, G. D., Gaudio, S. D., Galderisi, U., Cascino, A. and Cipollaro, M., Comparative evaluation of different DNA extraction procedures from food samples. Biotechnol. Prog., 2007, 23, 297-301.

  • Wilfinger, W. W., Mackey, K. and Chomczynski, P., Effect of pH and ionic strength on the spectrophotometric assessment of nucleic acid purity. BioTechniques, 1997, 22, 474-481.


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  • A Modified Protocol for Total Rna Isolation from Different Oil Palm (Elaeis guineensis) Tissues using Cetyltrimethylammonium Bromide

Abstract Views: 448  |  PDF Views: 124

Authors

Rashad Qadri
Hainan Key Laaboratory of Tropical Oil Crops Biology/Coconut Research Institute, Chinese Academy of Tropical Agricultural Science, Wenchang, Hainan 571339, China
Amjad Iqbal
Hainan Key Laaboratory of Tropical Oil Crops Biology/Coconut Research Institute, Chinese Academy of Tropical Agricultural Science, Wenchang, Hainan 571339, China
Yi Wu
Hainan Key Laaboratory of Tropical Oil Crops Biology/Coconut Research Institute, Chinese Academy of Tropical Agricultural Science, Wenchang, Hainan 571339, China
Jing Li
Hainan Key Laaboratory of Tropical Oil Crops Biology/Coconut Research Institute, Chinese Academy of Tropical Agricultural Science, Wenchang, Hainan 571339, China
Numrah Nisar
Department of Environmental Sciences, Lahore College for Women University Lahore, Lahore 54600, Pakistan
Muhammad Azam
Institute of Horticultural Sciences, University of Agriculture, Faisalabad, Pakistan
Yaodong Yang
Hainan Key Laaboratory of Tropical Oil Crops Biology/Coconut Research Institute, Chinese Academy of Tropical Agricultural Science, Wenchang, Hainan 571339, China

Abstract


Extraction of high-quality RNA from oil palm tissues is challenging due to the presence of polysaccharides, polyphenols and other complexes that co-precipitate with RNA. Therefore, isolation of high-quality RNA from oil palm is challenging due to the presence of varying amounts of these constituents in diverse tissues. This communication describes a modified RNA extraction protocol based on the cetyltrimethylammonium bromide (CTAB) method which is useful for extracting high-quality RNA from different oil palm tissues. Total RNA isolation using a modified CTAB protocol was compared with two different methods, a conventional TRIzol method and the method for RNA isolation from palms (MRIP). Both methods were useful for isolating RNA from leaf tissues; however, they were not effective in isolating RNA from other tissues. The current protocol based on a modified CTAB method was efficient in isolating high-quality total RNA from oil palm fruit tissues, including mesocarp and endosperm, stem, ischolar_main and flower tissues. This modified CTAB-based protocol gave approximately 20-30 μg of total RNA from 150 mg of tissue within 5-6 h.

Keywords


CTAB Method, Gene Expression, Oil Palm, RNA Extraction.

References





DOI: https://doi.org/10.18520/cs%2Fv116%2Fi3%2F479-482