Revista Chapingo Serie Ciencias Forestales y del Ambiente
Universidad Autónoma Chapingo
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Revista Chapingo Serie Ciencias Forestales y del Ambiente
Volume XVIII, issue 1, January - April 2012
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OPTIMIZACIÓN DE UN PROTOCOLO DEL AISLAMIENTO DEL ADN Y DE UN SISTEMA DE AMPLIFICACIÓN ISSR-PCR PARA Ceratozamia mexicana Brongn. (Zamiaceae).
OPTIMIZATION OF A PROTOCOL FOR DNA ISOLATION AND ISSR-PCR AMPLIFICATION SYSTEM FOR Ceratozamia mexicana Brongn. (Zamiaceae).

Nadia Guadalupe Sánchez-Coello; Mauricio Luna-Rodríguez; Santiago Mario Vázquez-Torres; Lázaro Rafael Sánchez-Velásquez; Nancy Santana-Buzzy; Pablo Octavio-Aguilar; Lourdes Georgina Iglesias-Andreu

http://dx.doi.org/10.5154/r.rchscfa.2011.03.024

Received: 2011-03-16

Accepted: 2011-11-01

Available online: / pages.123-133

 

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  • descriptionAbstract

    Most of the cycads contain high concentrations of essential oils, flavonoids, polyphenols, and polysaccharides that interfere with DNA extraction, causing erroneous or no PCR products. The optimization of DNA isolation, employing inter-simple sequence repeats (ISSRs) primers were investigated in Ceratozamia mexicana Brongn., an endangered Mexican cycad. The DNA obtained from fresh-leaf tissues with a modified cetyltrimethylammonium bromide buffer protocol gave a good quality of DNA with no colored pigments and contaminants. The main modification to the CTAB-based DNA extraction protocol was the one hour leaf tissue soaking pre-treatment with a 0.7 M NaCl solution, to facilitate the cell lysis. The DNA extracted was successfully amplified by PCR using six arbitrary ISSR primers. Reproducible amplifiable products were observed in all PCR reactions. Our results show a significant improvement in the DNA quality obtained using low primer concentration (25 pM). 23 strong bands were detected, 9 of which were polymorphic. The results indicated that the optimized protocol for DNA isolation and PCR system is suitable for further work in this specie. This work is the first DNA extraction and ISSR protocols reported for this ornamental and endangered species.

    Keyworks: Ceratozamia mexicana, Cycads, DNA extraction, endangered species, ISSR.
  • beenhereReferences
    • Aljanabi, S., Forget, L. & Dookun, A. (1999). An improved rapid protocol for the isolation of polysaccharide and polyphenol- free sugarcane DNA. Plant Molecular Biological Report, 17, 1-8.

    • Avise, J. C. (1994). Molecular Markers, Natural History, and Evolution. New York. Chapman & Hall.

    • Boiteux, L. S., Fonseca, M. & Simon, P. W. (1999). Effects of Plant Tissue and DNA Purification Method Randomly Amplified Polimorphic DNA-based Genetic Fingerprinting Analysis in Carrot. American Society of Horticulture Science, 124, 32-38.

    • Clark, M.S. (1997). Plant Molecular Biology, a Laboratory Manual. Berlin. Springer-Verlag.

    • Dellaporta, S. L., Wood, J. & Hocks, J. B. (1983). A plant DNA minipreparation: version II. Plant Molecular Biology Report, 1, 19-21.

    • Deshmukh, A. S., Treeback, J. T., Long, Y. C., Viollet, B., Wojtaszewski, J., F. & Zierat, H. J. R. (2007). Role of adenosine 5’-monophosphate-activated protein kinase subunits in skeletal muscle mammalian target of rapamycin signaling. Molecular Endocrinology, 22(5), 1105-1112.

    • Doyle, J. J. & Doyle J. L. (1990). A rapid DNA isolation procedure from small quantities of fresh leaf tissues. Phytochemical Bulletin, 19, 11-15.

    • Fang, G., Hammar, S. & Rebecca, R. (1992). A quick and inexpensive method for removing polysaccharides from plant genomic DNA. Biotechniques, 13, 52-56.

    • Flachowsky, H., Schumann, E., Weber, W. E. & Peil, A. (2001). Application of AFLP for the detection of sex-specific markers in hemp. Plant Breeding, 120, 305-309.

    • González-Astorga, J., Vovides, A. P., Octavio-Aguilar, P., Aguirre- Fey, D., Nicolalde-Morejon, F. & Iglesias, C. (2006). Genetic diversity and structure of the cycad Zamia loddigesii Miq. (Zamiaceae): implications for evolution and conservation. Botanical Journal of the Linnean Society, 152, 533-544.

    • Gangopadhyay, G., Roy, R. S., Ghose, K., Poddar, R., Bandyopadhyay, T., Basu, D. &, Mukherjee, K. K. (2007). Sex detection of Carica papaya and Cycas circinalis in pre-flowering stage by ISSR and RAPD. Current Science, 92(4), 524-526.

    • Jianguang, X., Shuguang, J. & Nian, L. (2005). Genetic variation in the endemic plant Cycas debaoensis on the basis of ISSR analysis. Australian Journal of Botany, 53, 141-145.

    • Katterman F. & Shattuck, V. L. (1983). An effective method of DNA isolation from the mature leaves of Gossypium species that contain large amounts of phenolic terpenoids and tannins. Preparative Biochemical, 13, 347-359.

    • Lodhi, M. A., Ye, G. N., Weeden, N. F. & Reisch, B. I. (1994). A simple and efficient method for DNA lysis from grapevine cultivars and Vitis species. Plant Molecular Biological Report, 12, 6-13.

    • Luna-Rodríguez, M., López-Upton, J. & Iglesias-Andreu L. G. (2005). Variabilidad morfométrica y molecular (RAPD) en una plantación de Pinus patula en Veracruz, México. Agrociencia, 39, 231-235.

    • Michelmore, R. W., Paran, I. & Kesseli, R. V. (1991). Identification of markers linked to disease resistance genes by bulked segregant analysis: A rapid method to detect markers in specific genome regions by using segregating populations. Proceedings of the National Academic of Science, 88, 9828-9832.

    • Murray, M. G. & Thompson, W. F. (1980). Rapid isolation of high molecular weight DNA. Nucleic Acids Research, 8, 4321-4325.

    • Octavio-Aguilar, P., González-Astorga,J. & Vovides, A. P. (2009). Genetic diversity through life history of Dioon edule Lindley (Zamiaceae, Cycadales). Plant Biology, 11, 525-536.

    • Prakash, S. & Staden, J. (2006). Sex identification in Encephalartos natalensis (Dyer and Verdoorn) using RAPD markers. Euphytica, 152(2),197-200.

    • Peng-Quiao, M. O., Yu-Yuan, H., Xiao-Qing, Z., Guang-Lin, L., Zheng-Wen, L. & Bao-Xuan, N. (2008). Establishment of Cycas micholitzii ISSR-PCR optimal conditions with orthogonal optimization method. Bulletin of Botanical Research, 28(3), 304-309.

    • Pollegioni, P., Major, A., Bartoli, S., Ducci, F., Proietti, R., Malvolti, M. E. & Anazato, D., (2006). Application of microsatellite and dominant molecular markers for the discrimination of species and interspecific hybrids in genus Juglans. Acta Horticulture, 705, 191-197.

    • Porebski, S., Bailey, L. G. & Baum, B. R. (1997). Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components. Plant Molecular Biological Report, 15, 8-15.

    • Reddy, G S. N., Prakash, J. S. S., Matsumoto, G. I., Stackebrandt, E. & Shivaji, S. (2002). Arthrobacter roseus sp. nov., a psychrophilic bacterium isolated from an Antarctic cyanobacterial mat sample. International Journal of Systematic Evolution and Microbiology, 52, 1017-1021.

    • Reamon-Büettner, S. M. & Jung, C. (2000). AFLP-derived STS markers for the identification of sex in Asparagus officinalis L. Theoretical and Applied Genetics, 100, 432-438.

    • Sarwat, M., Negi, M. S., Lakshmikumaran, M. & Tyagi, A. K. (2006). A standardized protocol for genomic DNA isolation from Terminalia arjuna for genetic diversity analysis. Electronic Journal of Biotechnology, 9, 86-91.

    • SEMARNAT (2010). Protección Ambiental de Especies Nativas de México de Flora y Fauna Silvestre. Categorías de Riesgo y Especificaciones para su Inclusión, Exclusión o Cambio. Lista de Especies en Riesgo. México. Secretaria del Medio Ambiente y Recursos Naturales.

    • Shah, M. M., Yen, Y., Gill, K. S. & Baenzinger, P. S. (2000). Com¬parisons of RFLP and PCR-based markers to detect polymorphism between wheat cultivars. Euphytica, 114, 135-142.

    • Sharma, K. K., Lavanya, M. & Anjaiah, V. (2000). A Method for isolation and purification of peanut genomic DNA suitable for analytical applications. Plant Molecular Biology Reporter, 18, 393a-393h.

    • Stewart, C. N. & Via, L. E. (1993). A rapid CTAB DNA isolation technique useful for RAPD fingerprinting and other PCR applications. BioTechniques, 14, 748-751.

    • Suman, P. S. K., Ajit, K. S., Darokar, M. P. & Kumar, S. (1999). Rapid isolation of DNA from dry and fresh samples of plants producing large amounts of secondary metabolites and essential oils. Plant Molecular Biology Report, 17, 1-7.

    • Vovides, A. P. (1983). Zamiaceae. In: V. Sosa, ed., Flora de Veracruz, Fasc. 26, 29. Xalapa, México. Instituto Nacional de Recursos Bióticos.

    • Vovides, A. P. & Iglesias, C. G. (1994). An intregrated conservation strategy for the cycad Dioon edule Lindl. Biodiversity and Conservation, 3, 137-141.

    • Warude, D., Chavan, P., Joshi, K. & Patwardhan, B. (2003). DNA isolation from fresh, dry plant samples with highly acidic tissue extracts. Plant Molecular Biological Report, 21, 467.

    • Yagi, F., Iwaya, T., Haraguchi, T. & Goldstein, I. J. (2002). The lectin from leaves of Japanese cycad, Cycas revoluta Thunb. (Gymnosperm) is a member of the jacalin-related family. European Journal Biochemistry, 269, 4335-4341.

    • Zietkiewicz, E., Rafalaski, A. & Labuda, D. (1994). Genome fingerprinting by simple sequence repeat (SSR) anchored polymerase chain reaction amplification. Genomics, 20, 176-183.

  • starCite article

    Sánchez-Coello, N. G.,  Luna-Rodríguez, M., Vázquez-Torres, S. M.,  Sánchez-Velásquez, L. R.,  Santana-Buzzy, N., Octavio-Aguilar, P., &  Iglesias-Andreu, L. G. (2012).  OPTIMIZATION OF A PROTOCOL FOR DNA ISOLATION AND ISSR-PCR AMPLIFICATION SYSTEM FOR Ceratozamia mexicana Brongn. (Zamiaceae).. Revista Chapingo Serie Ciencias Forestales y del Ambiente, XVIII(1), 123-133. http://dx.doi.org/10.5154/r.rchscfa.2011.03.024