The Cucurbita maxima Duchesne is a vegetable crop plant cultivated and maintained by traditional Amazon communities, Brazil. The situation is worsened by the possibility of disappearance of local populations and genetic variability of this specie, taking into account the today changes promoted in family farming. The aim of this study was to estimate the current levels of genetic variability of local cultivars through the use of molecular markers (Amplified Fragment Length Polymorphism—AFLP). We chose to collect in two distinct micro regions in order to identify possible influences of geographic isolation and different levels of market requirements in the conservation of the genetic variability of the C. maxima . For the molecular analysis, bulk samples of fresh leaves of 15 plants/half-sibling family were collected in paper bags. There were 34 samples from the half-sib families. The analysis of the results half-sib obtained by methods of estimation of genetic variation by molecular markers shows that the forms of cultivation and management adopted by family farmers maintain the identities of the local/landraces (native cultivars) and, at the same time, the levels of diversity for the assurance of adaptability macro-environmental.
KeywordsCrop Genetic VariabilityMolecular MarkerAmplified Fragment Length Polymorphism<I>Cucurbita maxima</I>Duchesne
Fonseca, M.A., Queiroz, M.A., Lopes, J.F., Assis, J.G.A., Romão, R.L., Ramos, S.R.R., Carmo, C.A.S., Moura, M.C.L., Dias, R.C.S., Silveira, L.M., Barbieri, R.L., Silva, D.J.H. and Noronha, S. (2015) Geographical Distribution and Conservation of Cucurbita in Brazil. Magistra, Cruz das Almas, BA, 27, 432-442.
Esquinas-Alcazar, J.T. and Gulick, P.J. (1983) Genetic Resources of Cucurbitaceae. IPBGR, Rome, 101.
Saade, R.L. and Hernández, S.M. (1992) La Agricultura em Mesoamerica: Cucurbitas. In: Hernández Bermejo, J.E. and Léon, J., Eds., Cultivos Marginados. Outra perspectiva de 1492. Colección FAO: Producción y protección vegetal, No. 26, Roma.
Heiden, G.; Barbieri, R. L.; Neitzke, R. S. (2007) Key for the identification of species of pumpkins (Cucurbita, Cucurbitaceae) grown in Brazil. (Documents, 197). Embrapa Temperate Weather, Pelotas, 31.
Canul Ku, J. Vallejo, P.R., González, F.C. and Servia, J.L.C. (2005) Morphological Diversity of Squash Grown in the Center-East of Yucatan, Mexico. Revista Fitotecnia Mexicana, 28, 339-349.
Carvalho, P.G.B., de Peixoto, A.A.P. and Ferreira, M.A.J.F. (2011) Characterization of Pumpkins as to the Levels of Total Carotenoids Alpha and Betacarotene (Research and Development Bulletin 78). Embrapa Hortaliças, Brasilia, 20.
Lima, G.K.L. (2013) Rescue and Germplasm Study of Cucurbita spp. of Rio Grande do Norte. Thesis, Mossoró.
OECD, Organization for Economic Co-operation and Development, Environment, Health and Safety Publications (2012) Consensus Document on the Biology of Cucurbita L. (Squashes, Pumpkins, Zucchinis and Gourds). Series on Harmonization of Regulatory Oversight in Biotechnology. No. 53 ENV/JM/MONO31, 77.
Bisognin, D.A. (2002) Origin and Evolution of Cultivated Cucurbits. Rural Science. Santa Maria, 32, 715-723. https://doi.org/10.1590/S0103-84782002000400028
Pasqual, M., Rezende, R.K.S., Villa, F. and Chagas, E.A. (2008) Biotechnology Applied to the Improvement of Fruit Trees. In: Bruckner, C.H., Ed., Fundamentals of Fruit Tree Breeding, UFV, Viçosa, 117-170.
Faleiro, F.G. (2007) Genetic-Molecular Markers Applied to Conservation and Use of Genetic Resources. Embrapa, Cerrados, Planaltina, 102.
Vos, P., Hogers, R., Bleeker, M., Reijans, M., van de Lee, M.T., Hornes, M., Frijters, A., Pot, J., Peleman, J., Kuiper, M. and Zabeau, M. (1995) AFLP: A New Technique for DNA Fingerprinting. Nucleic Acids Research, 23, 4407-4414. https://doi.org/10.1093/nar/23.21.4407
Caixeta, E.T., de Oliveira, A.C.B., de Brito, G.G. and Sakiyama, N.S. (2009) Types of Markers. In: Borém, A. and Caixeta, E.T. (Eds), Molecular Markers, Folha de Viçosa, Viçosa, 11-93.
Milach, S.C. K. (1998) DNA Markers. Applications in Plant Breeding. Biotechnology Science & Development, 5, 14-17.
Ferreira, M.E. and Grattapaglia, D. (1998) Introduction to the Use of Molecular Markers in Genetic Analysis. 3rd Edition, Document 20, Embrapa-Cenargen, Brasília, 220 p.
Brammer, S.P. (2000) Molecular Markers: Basic Principles and Use in Plant Breeding Programs. http://www.cnpt.embrapa.br/biblio/p_do03.htm
Milach, S.C.K. (1999) Molecular Markers on Genetic Resources and Plant Breeding. In: Queiroz, M.A., Goedert, C.O. and Ramos, S.R.R., Eds., Genetic Resources and Plant Breeding for the Brazilian Northeast, Embrapa Semiárido, Brasília, DF, Embrapa Genetic Resources and Biotechnology, Petrolina. http://www.cpatsa.embrapa.br:8080/catalogo/livrorg/index.html
Vencovsky, R. (1987) Effective Population Size in the Collection and Preservation of Germplasm Outcrossing Species. IPEF, 35, 79-84.
Martins, L.H.P. (2015) Genetic Variability and Conservation of Cucurbita maxima Duchesne by Family Agriculture in the Western Central Amazon. Thesis, Federal University of Amazonas, 151 p.
Couch, J.A. and Fritz, P.J. (1990) Isolation of DNA from Plants High in Polyphenolics. Plant Molecular Biology Reporter, 8, 8-12. https://doi.org/10.1007/BF02668875
Lopes, R., Lopes, M.T.G., Carneiro, M.S., Matta, F.P., Camargo, L.E.A. and Vieira, M.L.C. (2006) AFLP Linkage Analysis and Mapping of Resistance Genes to Xanthomonas axonopodis pv. passiflorae n Yellow Passion Fruit. Genome, 49, 17-29. https://doi.org/10.1139/g05-081
Creste, S., Tulmann Neto, A. and Figueira, A. (2001) Detection of Single Sequence Repeat Polymorphisms in Denaturing Polyacrylamide Sequencing Gels by Silver Staining. Plant Molecular Biology Reporter, 19, 299-306. https://doi.org/10.1007/BF02772828
Rohlf, F.J. (2000) NTSYS-pc, Version 2.1. Numerical Taxonomy and Multivariate Analysis System. Exeter Software, New York.
Hammer, Harper, D.A.T. and Ryan, P.D. (2012) PAST: Paleontological Statistics Software Package for Education and Data Analysis. Electronic Paleontology, 4, 187 .
Priori, D., Barbieri, R.L., Castro, C.M., Oliveira, A.C., Vilella, J.C.B. and Mistura, C.C. (2012) Molecular Characterization of Creole Varieties of Pumpkins with Microsatellite Markers. Brazilian Horticulture, 30, 499-506. https://doi.org/10.1590/S0102-05362012000300024
Restrepo, J.A. and Vallejo, C.F.A. (2008) Molecular Characterization of Colombian Introductions of Zapallo Cucurbita moschata. Acta Agronómica (Palmira), 57, 9-17.
Mingoti, S.A. (2005) Data Analysis through Multivariate Statistical Methods: An Applied Approach. Belo Horizonte. UFMG.
Clement, C.R., Rizzi Rocha, S.F., Cole, D.M. and Vivan, J.L. (2007) Conservation on Farm. In: Nass, L., Ed., Plant Genetic Resources, Embrapa Genetic Resources and Biotechnology, Brasília, 511-544.
Gwanama, C., Labuschagne, M.T. and Botha, A.M. (2000) Analysis of Genetic Variation in Cucurbita moschata by Random Amplified Polymorphic DNA (RAPD) Markers. Euphytica, 113, 19-24. https://doi.org/10.1023/A:1003936019095
Ferriol, M.B.P., Cordova, P.F. and Nuez, F. (2004) Molecular Diversity of a Germplasm Collection of Squash (Cucurbita moschata) Determined by SRAP and AFLP Markers. Crop Science, 44, 654-663.
Watcharawongpaiboon, N. and Chunwongse, J. (2007) Development of Microsatellite Markers from an Enriched Genomic Library of Pumpkin (Cucurbita moschata L.). Songklanakarin Journal of Science and Technology, 29, 1217-1223.