Revista de Biología Tropical ISSN Impreso: 0034-7744 ISSN electrónico: 2215-2075

Estimation of genetic variation in closely related cycad species in Ceratozamia (Zamiaceae: Cycadales) using RAPDs markers
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Ceratozamia norstogii complex
genetic variation
founder effects
complejo de Ceratozamia norstogii
variación genética
efecto fundador

How to Cite

Pérez-Farrera, M. A., Vovides, A. P., González, D., López, S., Hernandez-Sandoval, L., & Martínez, M. (2017). Estimation of genetic variation in closely related cycad species in Ceratozamia (Zamiaceae: Cycadales) using RAPDs markers. Revista De Biología Tropical, 65(1), 305–319.


The Ceratozamia norstogii complex from Southern Mexico is made up of four closely related taxa and occurs in similar habitats (Quercus forest). All have linear-lanceolate leaflets with great similarity between them, especially in juvenile stages, but differentiate with age. There has been debate regarding delimitation of species due to character loss in herbarium specimens. The aim of this study was to determine the genetic variation, and to measure genetic similarity between the four taxa. We studied populations in Cintalapa (Chiapas) for C. alvarezii and C. norstogii; the Sierra Atravesada (Oaxaca) for C. chimalapensis, and Villa Flores (Chiapas) for C. mirandae. One population for each taxon was sampled (only one population is known for C. alvarezii) 11-15 randomly chosen adult individuals were sampled. Twenty-eight primers were tested of which five were polymorphic using the RAPD’S technique. The data were analyzed using Bayesian methods. Results revealed low genetic diversity, and a differentiation was found between species, suggesting a recent divergence. A previous morphological and anatomical study on the complex has found the taxa to be distinct. However, the results of this study have shown that the C. norstogii species complex is in a divergence process, probably through genetic drift and founder effects.
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Adam, R. P., Morris, J. A., Pandey, R. N., & Schwarzbach, A. E. (2005). Cryptic speciationbetween Juniperus deltoides and Juniperus oxycedrus (Cupressaceae) in the Mediterranean. Biochemical Systematics and Ecology, 33, 771-787.

Archak S., Karihaloo, J. L., & Jain, A. (2002). RAPD markers reveal narrowing genetic base of Indian tomato cultivars. Current Science, 82, 1139-1143.

Bruschi, P., Vendramin, G. G., Bussati, F., & Grossoni, P. (2003). Morphological and molecular diversity among Italian population of Quercus petraea (Fagaceae). Annals of Botany, 91, 707-716.

Burnham, R. J., & Graham, A. (1999). The history of neotropical vegetation: new developments and status. Annals of the Missouri Botanical Garden, 86, 546-589.

Cabrera-Toledo, D., González-Astorga, J., & Vovides, A. P. (2008). Heterozygote excess in ancient populations of the critically endangered Dioon caputoi (Zamiaceae, Cycadales) from central Mexico. Botanical Journal of the Linnean Society, 158, 436-447.

Cabrera-Toledo, D., González-Astorga, J., Nicolalde-Morejón, F., Vergara-Silva, F., & Vovides, A. P. (2010). Allozyme diversity levels in two congeneric Dioon spp. (Zamiaceae, Cycadales) with contrasting rarities. Plant Systematics and Evolution, 290, 115-125.

Carelli, B. P., Gerald, L. T. S., Grazziotin, F. G., & Echeverrigaray, S. (2006). Genetic diversityamong Brazilian cultivars and landraces of tomato Lycopersicon esculentum Mill. Revealed by RAPD markers. Genetic Resources and Crop Evolution, 53, 395-400.

Chaiprasongsuk, M., Tantivivat, S., Ming, M., Thongpan, A., Sangduan, N., Lindstrom, A. J., & Namwongprom, K. (2004). Phylogenetic relationships in Encephalartos (Zamiaceae) using random amplified polymorphic DNA (RAPD). In A. J. Lindstrom (Ed.), Proceedings of the Sixth International Conference on Cycad Biology: The Biology, Structure and Systematics of the Cycadales (pp. 37-43). Chonburi, Thailand: Nong Nooch Tropical Botanical Garden.

Chapco, W., Ashton, N. W., Martel, R. K. B., Antonishyn, N., & Crosb, W. L. (1992). A feasibilitystudy of the use of random amplified polymorphic DNA in the population genetics and systematics of grasshoppers. Genome, 35, 569-574.

Cheng, K. T., Chang, H. C., Su, C. H., & Hsu, F. L. (1997). Identification of dried rhizomes of Coptis species using random amplified polymorphic DNA. Botanical Bulletin of Academia Sinica, 38, 241-244.

Cibrián-Jaramillo, A., Marler, T. E., De Salle, R., & Brenner, E. D. (2008). Development of EST- microsatellites from the cycad Cycas rumphii, and their use in the recently endangered Cycas micronesica. Conservation Genetics, 9, 1051-1054.

Cibrián-Jaramillo, A., Daly, A. C., Brenner, E., De Salle, R., & Marler, T. E. (2010). When North and South don’t mix: genetic connectivity of a recently endangered oceanic cycad, Cycas micronesica, in Guam using EST‐microsatellites. Molecular Ecology, 19, 2364-2379.

Crawford, D. L. (1983). Phylogenetic and systematic inferences from electrophoretic studies. In S. D. Tanksley, & T. J. Orton (Eds.), Isozymes in plant genetics and breeding Part B (pp. 257-287). Amsterdam, The Netherlands: Elsevier.

De Castro, O., Vázquez-Torres, M., & De Luca, P. (2006). Utility of AFLP markers for the assessment of molecular relationships in Ceratozamia Brongn. (Zamiaceae). Plant Biosystems, 140, 221-228.

Demeke, T., Lynch, D. R., Kawchuk, L. M., Kozub, G. C., & Armstrong, J. D. (1996). Genetic diversity of potato determined by random amplified polymorphic DNA analysis. Plant Cell Reports, 15, 662-667.

Dudley, E. C. (1978). Adaptive radiation in the Melastomataceae along an altitudinal gradient in Peru. Biotropica, 10, 134-143.

Ellstrand, N. C., Ornduff, R., & Clegg, J. M. (1990). Genetic structure of the Australian cycad Macrozamia communis (Zamiaceae). American Journal of Botany, 77, 677-681.

Erlich, H. A., Gelfand, D., & Sninsky, J. J. (1991). Recent advances in the Polymerase Chain-Reaction. Science, 252, 1643-1651.

Ekué, M., Gailing, R. M., Hölscher, O., Sinsin, D. B., & Finkeldey, R. (2008). Population genetics of the cycad Encephalartos barteri ssp. barteri (Zamiaceae) in Benin with notes on leaflet morphology and implications for conservation. Belgian Journal of Botany, 141, 78-94.

Feng, X., Wang, Y., & Gong, X. (2014). Genetic diversity, genetic structure and demographic history of Cycas simplicipinna (Cycadaceae) assessed by DNA sequences and SSR markers. BMC Plant Biology, 14, 187-202.

Gepts, P. (1993). The use of molecular and biochemical markers in crop evolution studies. Evolutionary Biology, 27, 51-94.

González, D., & Vovides, A. P. (2002). Low intralineage divergence in the genus Ceratozamia Brongn. (Zamiaceae) detected with nuclear ribosomal DNA ITS and chloroplast DNA trnL-F non-coding region. Systematic Botany, 27, 654-661.

González-Astorga, J., Vovides, A. P., Ferrer, M. M., & Iglesias, C. (2003). Population genetics study in the Mexican endemic cycad Dioon edule Lindl. (Zamiaceae): Biogeographical and evolutionary implications. Biological Journal of the Linnean Society, 80, 457-467.

González-Astorga, J., Vovides, A. P., Cruz-Angón, A., Octavio-Aguilar, P., & Iglesias, C. (2005). Allozyme variation in the three extant populations of the narrowly endemic cycad Dioon angustifolium Miq. (Zamiaceae) from northeastern Mexico. Annals of Botany, 95, 999-1007.

González-Astorga, J., Vergara-Silva, F., Vovides, A. P., Nicolalde-Morejón, F., Cabrera-Toledo, D., & Pérez-Farrera, M. A. (2008). Diversity and genetic structure of three species of Dioon Lindl. (Zamiaceae, Cycadales) from the Pacific seaboard of Mexico. Biological Journal of the Linnean Society, 94, 765-776.

González-Astorga, J., Vovides, A. P., Cabrera-Toledo, D., & Nicolalde-Morejón, F. (2009). Diversity and genetic structure of the endangered cycad Dioon sonorense (Zamiaceae) from Sonora, Mexico: Evolutionary and conservation implications. Biochemical Systematics and Ecology, 36, 891-899.

Gottlieb, L. D. (1981). Electrophoretic evidence and plant populations. Progress in Phytochemistry, 7, 1-46.

Grant, V. (1985). The evolution process: A critical review of evolutionary theory. New York: Columbia University Press.

Hamrick, J. L. (2004). Response of forest trees to global environment changes. Forest Ecology and Management, 197, 323-335.

Hamrick, J. L., & Godt, M. J. (1989). Allozyme diversity in plant species. In A. H. D. Brown, M. T. Clegg, A. H. Kahler, & B. S. Weir (Eds.), Population genetic and germplasm resources in crop improvement genetic resources (pp. 43-63). Sunderland, England: Sinauer.

Huang, S., Chiang, Y. C., Schaal, B. A., Chou, C. H., & Chiang, T. Y. (2001). Organelle DNA phylogeography of Cycas taitungensis, a relict species in Taiwan. Molecular Ecology, 10, 2669-2681.

Keppel, G., Lee, S. W., & Hodgskiss, P. D. (2002). Evidence for long isolation among populations of a pacific cycad: genetic diversity and differentiation in Cycas seemannii A.Br. (Cycadaceae). Journal of Heredity, 93, 133-139.

Khan, A. I., Khan, I. A., Awan, F. S., Sadaqat, H. A., & Bahadur, S. (2011). Estimation of genetic distance based on RAPDs between 11 cotton accessions varying in heat tolerance. Genetics Molecular Research, 10, 96-101.

Landry, B. S., Dextraze, L., & Boivin, G. (1993). Random amplified polymorphic DNA markers for DNA fingerprinting and genetic-variability assessment of minute parasitic wasp species (Hymenoptera, Mymaridae and Trichogrammatidae) used in biological-control programs of phytophagous insects. Genome, 36, 580-587.

Long-Qian, X., Xue-Jun, G. J., Xun, G., Gang, H., & Si-Xiang, Z. (2004). ISSR variation in the endemic and endangered plant Cycas guizhouensis (Cycadaceae). Annals of Botany, 94, 133-138.

Lynch, M., & Milligan, B. G. (1994). Analysis of population genetic structure with RAPD markers. Molecular Ecology, 3, 91-99.

Meerow, A. W., &. Nakamura, K. (2007). Ten microsatellite loci from Zamia integrifolia (Zamiaceae). Molecular Ecology Notes, 7, 824-826.

Meerow, A. W., Francisco-Ortega, J., Calonje, M., Griffiths, M. P., Ayala-Silva, T., Stevenson, D. W., & Nakamura, K. (2012). Zamia (Cycadales: Zamiaceae) on Puerto Rico: Asymmetric genetic differentiation and the hypothesis of multiple introductions. American Journal of Botany, 99, 1828-1839.

Mekanawakul, M., & Juntawong, N. (2004). Diversity and genetic variation among populations of Thai cycads revealed by ALFP markers. In A. J. Lindstrom (Ed.), Proceedings of the sixth International conference on cycad biology. The biology, structure and systematics of the Cycadales (pp. 44-59). Chonburi, Thailand: Nong Nooch Tropical Botanical Garden.

Miller, M. P. (1997). Tool for population genetic analyses (TFPGA) 1.3: A windows program for the analysis of allozyme and molecular population genetic data. Computer software distributed by author.

Moretti, A., Sabato, S., & Vázquez-Torres, M. (1980). The distribution of Ceratozamia (Zamiaceae). Delpinoa, 20, 13-21.

Moynihan, J., Meerow, A. W., & Francisco-Ortega, J. (2007). Isolation, characterization and cross‐ species amplification of microsatellite loci in the cycad genus Dioon (Zamiaceae). Potential utilization in population genetics studies of Dioon edule. Molecular Ecology Notes, 7, 72-74.

Mullis, K. B. (1990). The unusual origin of the Polymerase Chain-Reaction. Scientific American, 262, 56-64.

Norstog, K. J., & Nicholls, T. J. (1997). The biology of cycads. Ithaca: Cornell University Press.

Norstog, K. J. (1987). Cycads and the origin of insect pollination. American Scientist, 75, 270-279.

Norstog, K., & Fawcett, P. K. S. (1989). Insect-cycad symbiosis and its relation to the pollination of Zamia furfuracea (Zamiaceae) by Rhopalotria mollis (Curculionidae). American Journal of Botany, 76, 1380-1394.

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.

Osborne, R., Calonje, M., Hill, K., Stanberg, L., & Stevenson, D. W. (2012). The world list of cycads. Memoirs of the New York Botanical Garden, 106, 480-508.

Peakall, R., & Smouse, P. E. (2006) GENALEX 6: genetic analysis in Excel. Population genetic software for teaching and research. Molecular Ecology Notes, 6, 288-295.

Pérez-Farrera, M. A., Vovides, A. P., & Iglesias, C. (1999). A new species of Ceratozamia (Zamiaceae, Cycadales) from Chiapas, México. Novon, 9, 410-413.

Pérez-Farrera, M. A., Vovides, A. P., & Iglesias, C. (2001). The cycad Ceratozamia norstogii D.W.Stev. (Zamiaceae) from southern Mexico: new information on distribution, habitat and vegetative morphology. Botanical Journal of the Linnean Society, 137, 71-7.

Pérez-Farrera, M. A., Vovides, A. P., Hernández-Sandoval, L., González, D., & Martínez, M. (2004). A morphometric analysis of the Ceratozamia norstogii complex (Zamiaceae). In T. Walters & R. Osborne (Eds.), Cycad classification: concepts and recommendations (pp. 127-136). Wallingford, England: CABI Publishing.

Pérez-Farrera, M. A., Vovides, A. P., & Avendaño, S. (2014). Morphology and leaflet anatomy of the Ceratozamia norstogii (Zamiaceae, Cycadales) species complex in Mexico with comments on relationships and speciation. International Journal of Plant Science, 175, 110-121.

Pinares, A., González-Astorga, J., Vovides, A. P., Lazcano, J., & Vendrame, W. A. (2009). Genetic diversity of the endangered endemic Microcycas calocoma (Miq.) A. DC. (Zamiaceae, Cycadales): Implications for conservation. Biochemical Systematics and Ecology, 37(4), 385-394.

Qing-Qing, Z., Wang, J. F., Gong, X., & Peng, H. (2011). Patterns of chloroplast DNA variation in Cycas debaoensis (Cycadaceae): conservation implications. Conservation Genetics, 12, 959-970.

Raynolds, J., Weir, B. S., & Cockerham, C. C. (1983). Estimation of the co-ancestry coefficient: basis for a short-term genetic distance. Genetics, 105, 767-779.

Sangin, P., Thongpan, A., Lindstrom, A. J., Sangduan, N., & Mingmuang, M. (2004). Identification of Cycas by restricted fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD). In A. J. Lindstrom (Ed.), Proceedings of the sixth International conference on cycad biology. The biology, structure and systematics of the Cycadales (pp. 30-36). Chonburi, Thailand: Nong Nooch Tropical Botanical Garden.

Schneider, C. J., Smith, T. B., Larison, B., & Moritz, C. (1999). A test of alternative model of diversification in tropical forest: ecological gradients vs. rainforest refugia. Proceedings of the National Academy of Science, USA 96, 13869-13873.

Sharma, I. K., Jones, D. L., Forster, P. I., & Young, A. G. (1998). The extent and structure of genetic variation in the Macrozamia pauli-guilielmi complex (Zamiaceae). Biochemical Systematics and Ecology, 26, 45-54.

Sharma, I. K., Jones, D. L., Forster, P. I., & Young, A. G. (1999). Low isozymic differentiation among five species of the Macrozamia heteromera group (Zamiaceae). Biochemical Systematics and Ecology, 27, 67-77.

Sharma, I. K., Jones, D. L., & Forster, P. I. (2004). Genetic differentiation and phenetic relatedness among seven species of the Macrozamia plurinervia complex (Zamiaceae). Biochemical Systematics and Ecology, 32, 313-327.

Sokal, R. R., & Rohlf, F. J. (1985). Biometry 3rd ed. San Francisco: Freeman.

Stevenson, D. (1982). A new species of Ceratozamia (Zamiaceae) from Chiapas, México. Brittonia, 34, 181-184.

Stevenson, D. W., Sabato, S., & Vázquez Torres, M. (1986). A new species of Ceratozamia (Zamiaceae) from Veracruz, México with comments on species relationships, habitats, and vegetative morphology in Ceratozamia. Brittonia, 38, 17-26.

Van der Bank, H., Wink, M., Vorster, P., Treutlein, J., Brand, L., Van der Bank, M., & Hurter, J. (2001). Allozyme and DNA sequence comparisons of nine species of Encephalartos (Zamiaceae). Biochemical Systematics and Ecology, 29, 241-266.

Vekemans, X., Beauwens, T., Lemaire, M., & Roldán-Ruiz, I. (2002). Data from amplified fragment length polymorphism (AFLP) markers show indication of size homoplasy and of a relationship between degree of homoplasy and fragment size. Molecular Ecology, 11, 139-151.

Vovides, A. P., Rees, J. D., & Vázquez-Torres, M. (1983). Zamiaceae. In A. Gómez-Pompa & V. Sosa (Eds.), Flora de Veracruz (Fasc. 26, pp. 1-31). Xalapa, Mexico: INIREB.

Vovides, A. P., Pérez-Farrera, M. A., & Iglesias, C. (2001). Another new species of Ceratozamia (Zamiaceae) from Chiapas, México. Botanical Journal of the Linnean Society, 137, 81-85.

Vovides, A. P., Pérez-Farrera, M. A., González-Astorga, J., & Iglesias, C. (2008). A new species of Ceratozamia (Zamiaceae) from Oaxaca, Mexico with comments on habitat and relationships. Botanical Journal of the Linnean Society, 157, 169-175.

Vovides, A. P., Avendaño, S., Pérez-Farrera, M. A., & Stevenson, D. W. (2012). What is Ceratozamia brevifrons (Zamiaceae)? Brittonia, 64, 35-42.

Walters, T. W., & Decker-Walters, D. S. (1991). Patterns of allozyme diversity in the West Indies cycad Zamia pumila (Zamiaceae). American Journal of Botany, 78, 436-445.

Welsh, J., & McClelland, M. (1990). Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Research, 18, 7213-7218.

Whitelock, L. (2002). The Cycads. Portland, USA: Timber Press.

Williams, J. G. K., Kubelik, A. R., Livak, K. J., Rafalski, J. A., & Tingey, S. V. (1990). DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research, 18, 6531-6535.

Wright, S. (1943). Isolation by distance. Genetics, 28, 114-138.

Yang, S. L., & Meerow, A. W. (1996). The Cycas pectinata (Cycadaceae) complex: genetic structure and gene flow. International Journal of Plant Sciences, 157, 468-483.

Zhivotovsky, L. A. (1999). Estimating population structure in diploids with multi-locus dominant DNA markers. Molecular Ecology, 8, 907-913.


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