Análisis de la Variabilidad Genética entre treinta accesiones de tarwi (Lupinus mutabilis Sweet) usando marcadores moleculares ISSR

Michelle C. Chirinos-Arias, Jorge E. Jiménez, Lizbeth S. Vilca-Machaca

Resumen


Con el fin de realizar el análisis de variabilidad genética inter-accesión de treinta accesiones de tarwi (L. mutabilis Sweet) pertenecientes al Banco de Germoplasma del Instituto Nacional de Innovación Agraria (INIA). Se extrajo el ADN de 300 plantas, se construyeron bulks, se estandarizó el protocolo de amplificación de los marcadores moleculares Inter Simple Sequence Repeat (ISSR), de los cuales se eligió a los más polimórficos y nítidos para corrida en gel de acrilamida. Encontrándose 255 bandas con 8 iniciadores ISSR. El análisis de la variabilidad genética con estos iniciadores comprobó una alta variabilidad genética de las muestras en estudio. Observándose también un polimorfismo relativamente alto para una especie autógama como L. mutabilis. Finalmente los fenogramas mostraron una relación con la ubicación geográfica, posiblemente debido al flujo génico in situ debido al intercambio o venta de semillas en ferias o mercados aledaños a la zona de colecta.

Palabras clave


Lupinus mutabilis, bulks, variabilidad genética, ISSR

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Referencias


Aguilera, J.G.; Pessoni, L.A.; Rodrigues, G.B., Elsayed, A.Y.; Da Silva, D.J.; De Barros, E.G. 2011. Genetic variability by ISSR markers in tomato (Solanum lycopersicon Mill). Revista Brasileira Ciencias Agrarias 6: 243-252.

Astarini, I.A.; Plummer, J.A.; Lancaster, R.A.; Yan, G. 2006. Genetic diversity of Indonesian cauliflower cultivars and their relationships with hybrid cultivars grown in Australia. Scientia Horticulturae 108: 143-150.

Azofeifa, A. 2006. Uso de marcadores moleculares en plantas: aplicaciones en frutales del trópico. Agronomía Mesoamericana 17: 221-242.

Bornet, B.; Branchard, M. 2001. Nonanchored inter simple sequence repeat (ISSR) markers: reproducible and specific tools for genome fingerprinting. Plant Molecular Biology Reporter 19: 209–215.

Bussell, J.D.; Waycott, M.; Chappill, J.A. 2005. Arbitrarily amplified DNA markers as characters for phylogenetic. Perspectives in Plant Ecology, Evolution and Systematics 7: 3-26.

Camarena F, Huaringa A, Jiménez J, Mostacero E. 2012. Revalorización de un cultivo subutilizado: Chocho o Tarwi (Lupinus mutabilis Sweet). Editorial CONCYTEC, Lima, Perú.

Cao, P.Y.; Yao, Q.F.; Ding, B.Y.; Zeng, H.Y.; Zhong, Y.X.; Fu, C.X.; Jin, X.F. 2006. Genetic diversity of Sinojackia dolichocarpa (Styracaceae), a species endangered and endemic to China, detected by inter-simple sequence repeat (ISSR). Biochemical Systematics and Ecology 34: 231-239.

Castañeda, B.; Manrique, R.; Gamarra, F.; Muñoz, A.; Ramos, F.; Lizaraso, F.; Martínez, J. 2008. Probiótico elaborado en base a las semillas de Lupinus mutabilis Sweet. Acta Médica Peruana 25: 210-215.

Castañeda, B.; Castro de la Mata, R.; Manrique, E.; Ibañez, L. 2007. Efectos metabólicos de Lepidium meyenii Walpers.“MACA” y Lupinus mutabilis Sweet. “chocho” en ratas. Revista Horizonte Médico 7: 32-38.

Ciancio, A.; Mukerji, K.G. 2008. Integrated Management and Biocontrol of Vegetable and Grain Crops Nematodes 2: 102-103

Doyle, J.J.; Doyle, J.L. 1987. A rapid DNA isolation procedure from small quantities of fresh leaf tissues. Phytochem Bull. 19:11-15.

Eguiarte, L.E.; Souza, V.; Aguilar, X. 2007. Ecología Molecular. Editorial SEMARNAT. México.

FAOa. Org. Comunicado de prensa: 925 millones de personas sufren hambre crónica en el mundo. Disponible en: http://www.fao.org/news/story/es/item/45291/icode/

FAOb. Org. Cultivos Andinos, tarwi o chocho (Lupinus mutabilis). Disponible en: http://www.rlc.fao.org/es/agricultura/produ/cdrom/contenido/libro10/cap03_1_3

Ferriol, M.; Picó, B.; Nuez, F. 2001. Genetic Variability in Pumpkin (Cucurbita maxima) Using RAPD Markers. Cucurbit Genetics Cooperative Report 24: 94-96.

Fikiru, E.; Tesfaye, K.; Bekele, E. 2007. Genetic diversity and population structure of Ethiopian lentil (Lens culinaris Medikus) landraces as revealed by ISSR marker. African J. of Biotechnology 6: 1460-1468.

Fisher, P.J.; Gardner, R.C. y Richardson, T.E. 1996. Single locus microsatellites isolated using 5’anchored PCR. Nucleic Acids Research 24: 4369-4371.

Gilbert, J.E.; Lewis, R.V.; Wilkinson, M.J.; Caligari; P.D.S. 1999. Developing an appropriate strategy to assess genetic variability in plant germplasm collections. Theor. Appl. Genet. 98: 1125-1131.

Girma, G.; Tesfaye, K.; Bekele, E. 2010. Inter Simple Sequence Repeat (ISSR) analysis of wild and cultivated rice species from Ethiopia. African Journal of Biotechnology 9: 5048-5059.

Gross, R.; von Baer, E.; Koch, F.; Marquard, R.; Trugo, L.; Wink, M. 1988. Chemical composition of a new variety of the Andean lupin (Lupinus mutabilis cv. Inti) with low alkaloid content. J. Food Comp. Anal. 1: 353-361.

Guo, H.B.; Huang, K.Y.; Zhou, T.S.; Wu, Q.H.; Zhang, Y.J.; Liang, Z.S. 2009. DNA isolation, optimization of ISSR-PCR system and primers screening of Scutellaria baicalensis. Journal of Medicinal Plants Research 3(11): 898-901.

Hatcher, P.E.; Wilkinson, M.J.; Albani, M.C.; Hebbern, C.A. 2004. Conserving marginal populations of the food plant (Impatiens noli-tangere) of an endangered moth (Eustroma reticulatum) in a changing climate. Biological Conservation 116: 305-317.

Hollingsworth, P.M.; Ennos, R.A. 2004. Neighbour joining trees, dominant markers and population genetic structure. Heredity 92: 490-498.

Horovitz, A.; Harding, J. 1983. Genetics of Lupinus. XII. The mating system of Lupinus pilosus. Bot. Gaz. 144: 276-279.

Hou, Y.C.; Yan, Z.H.; Wei, Y.M.; Zheng, Y.L. 2005. Genetic diversity in barley from west China based on RAPD and ISSR analysis. Barley Genetics Newsletter 35: 9-22.

Isshiki, S.; Iwata, N., Mizanur, M.D. 2008. ISSR variations in eggplant (Solanum melongena L.) and related Solanum species. Scientia Horticulturae 117: 186-190.

Jacobsen, E.; Mujica, A. 2006. El tarwi (Lupinus mutabilis Sweet) y sus parientes silvestres. Universidad Mayor de San Andrés, La Paz, Bolivia. Pp. 458-482.

Jian, S.; Tang, T.; Zhong, Y.; Shi, S. 2004. Variation in inter-simple sequence repeat (ISSR) in mangrove and non-mangrove populations of Heritiera littoralis (Sterculiaceae) from China and Australia. Aquatic Botany 79: 75-86.

Jiménez, J. 2006. Biodiversity of traditional seed propagated crops cultivated in Peruvian highland. Tesis Ph.D. University of SILESIA. Polonia.

Joshi, N.; Rawat, A.; Subramanian, R.B.; Rao, K.S. 2010. A method for small scale genomic DNA isolation from chickpea (Cicer arietinum L.) suitable for molecular marker analysis. Indian Journal of Science and Technology 3: 0974- 6846.

Kameli, M.; Hesamzadeh, S.M.; Ebadi, M. 2013. Assessment of genetic diversity on populations of three satureja species in Iran using ISSR markers. Annals of Biological Research 4:64-72 63.

Konate, I.; El-Bekkay, B.; Filali-Maltouf, A. 2009. Inter-simple sequence repeat markers variation among natural accessions of Moroccan carob tree (Ceratonia siliqua). Int. J. Agric. Biol. 11: 168–172.

Kothera, L.; Ward, S.M.; Carney, S.E. 2007. Assessing the threat from hybridization to the rare endemic Physaria belli Mulligan (Brassicaceae). Biological Conservation 140: 110-118. 65.

Lázaro, A.; Aguinagalde, I. 2006. Genetic variation among Spanish pea landraces revealed by Inter Simple Sequence Repeat (ISSR) markers: its application to establish a core collection. Journal of Agricultural Science 144: 53–61.

Liu, L.W.; Zhao, L.P; Gong, Y.Q.; Wang, M.X.; Chen, L.M.; Yang, J.L; Wang, Y.; Yu, F.M.; Wang, L.Z. 2008. DNA fingerprinting and genetic diversity analysis of late-bolting radish cultivars with RAPD, ISSR and SRAP markers. Sci. Hort. 116: 240- 247.

Loarce, Y.; Gallego, R.; Ferrer, E. 1996. A comparative analysis of the genetic relationships between rye cultivars using RFLP and RAPD markers. Euphytica 88: 107-115.

Moreno, Y.; Melgarejo, L.M; Hernández, M.S.; Quintero, L.; Vargas, G. 2004. Caracterización molecular de un banco de germoplasma del género Theobroma mediante marcadores RAPD. Revista Colombiana de Biotecnología 4(2): 15-24.

Mujica, A. 1990. Investigación y producción del tarwi (Lupinus mutabilis Sweet.) en el Perú. INIAA, PICA. Lima.

Muthusamy, S.; Kanagarajan, S., Ponnusamy, S. 2008. Efficiency of RAPD and ISSR markers system in accessing genetic variation of rice bean (Vigna umbellata) landraces. Electronic Journal of Biotechnology 11: 0717-3458.

Nascimiento, M.A.; Batalha-Filho, H.; Waldschmidt, A.M.; Tavares, M.G.; Campos, L.A.O; Salomão, T.M. 2010. Variation and genetic structure of Melipona quadrifasciata Lepeletier (Hymenoptera, Apidae) populations base on ISSR pattern. Genetics and Biology 33(2): 394-397.

Ovesná, J.; Poláková, K.; Leisová, L. 2002. DNA Analyses and their Applications in Plant Breeding. Czech J. Genet. Plant Breed. 38: 29-40.

Palacios, A. 2003. Obtención de alcohol a partir de la malta de Lupinus mutabilis (Tarwi). Facultad de Ingeniería química. Universidad Nacional del Centro del Perú.

Rakoczy M.; Bolibok H. 2004. Characteristics and comparison of three classes of microsatellite-based markers and their application in plants. Cellular and molecular Biology Letters 9: 231-238.

Roh, M.S.; Cheong, E.J.; Choi, I.K.; Joung, Y.H. 2007. Characterization of wild Prunus yedoensis analyzed by inter-simple sequence repeat and chloroplast DNA. Scientia Horticulturae 114: 121-128.

Sbabou, L.; Brhada, F.; Alami, I.T. 2010. Genetic Diversity of Moroccan lupinus Germplasm Investigated using ISSR and AFLP Markers. International Journal of Agricultural & Biology 12: 26-32.

Sevilla, R.; Holle, M. 2004. Formas de reproducción de las plantas y Caracterización del germoplasma. Editorial Recursos genéticos vegetales Luis león Asociados S. R. L., Lima, Perú.

Shafie, M.; Hasan, S., Zain, A.; Shah, R. 2011. RAPD and ISSR markers for comparative analysis of genetic diversity in wormwood capillary (Artemisia capillaris) from Negeri Sembilan, Malaysia. J. Med. Plant. Res. 5(18): 4426-4437.

Sica, M.; Gamba, G.; Montieri, S.; Gaudio, L.; Aceto, S. 2005. ISSR markers show differentiation among Italian populations of Asparagus acutifolus L. BMC Genet. 6: 17

Su, Y.J.; Zan, Q.J.; Wang, T.; Ying, Z.M.; Ye, H.G 2008. High ISSR variation in 24 surviving individuals of Apterosperma oblata (Theaceae) endemic to China. Biochemical Systematics and Ecology 36: 619-625.

Svetleva, D.; Pereira, G.; Carlier, J.; Cabrita, L.; Leitão, J.; Genchev, D. 2006. Molecular characterization of Phaseolus vulgaris L. genotypes included in Bulgarian collection by ISSR and AFLP analyses. Sci Hort 109: 198-206.

Talhinhas, P.; Neves-Martins, J.; Leitao, J. 2003. AFLP, ISSR and RAPD markers reveal high levels of genetic diversity among Lupinus spp. Plant Breeding 122: 507-510.

Venkatachalam, L.; Sreedhar, R.V.; Bhagyalakshmi, N. 2008. The use of genetic markers for detecting DNA polymorphism, genotype identification and phylogenetic relationships among banana cultivars. Molecular Phylogenetics and Evolution 47: 974-985.

Vuylsteke, M.; Mank, R.; Brugmans, B.; Stam, P.; Kuiper, M. 2000. Further characterization of AFLP data as a tool in genetic diversity assessments among maize (Zea mays) inbred lines. Mol. Breed. 6: 265-276.

Wang, C.; Zhang, H.; Qian, Z.Q.; Zhao, G.F. 2008. Genetic differentiation in endangered Gynostemma pentaphyllum (Thunb.) Makino based on ISSR polymorphism and its implications for conservation. Biochemical Systematics and Ecology 36: 699-705.

Weising, K.; Nybom, H., Pfenninger, M.; Wolff, K.; Kahl, G. 2005. DNA fingerprinting in Plants. Principles, Methods, and Applications. Taylor and Francis Group, Boca Ratón. Pp 444.

Xu, F.; Sun, M. 2001. Comparative analysis of phylogenetic relationships of grain Amaranths and their wild relatives (Amaranthus; Amaranthaceae) using internal transcribed spacer, Amplified Fragment Length Polymorphism and Double-primer Fluorescent Intersimple Sequence Repeat Markers. Molecular Phylogenetics and Evolution 21: 372-387.

Yang, X.; Quiros, C. 1993. Identification and classification of celery cultivars with RAPD markers. Theor. Appl. Genet. 86: 205-212.

Zietkiewicz, E.; Rafalski, A.; Labuda, D. 1994. Genome fingerprinting by simple sequence repeats (SSR)-anchored polymerase chain reaction amplification. Genomics 20: 176–183.

Zhao, J.; Wang, X.; Deng, B.; Lou, P.; Wu, J.; Sun, R.; Xu, Z.; Vromans, J.; Koornneef, M.; Bonnema, G. 2005. Genetic relatioships within Brassica rapa as inferred from AFLP fingerprints. Theor. Appl. Genet. 110: 1301-1314.

Zhu, Y.; Hu, J.; Han, R.; Wang, Y.; Zhu, S. 2011. Fingerprinting and identification of closely related wheat (Triticum aestivum L.) cultivars using ISSR and fluorescence-labeled TP-M13-SSR markers. Australian Journal of Crop Science 5: 846-850.

Received: 10/09/14; Accepted: 14/02/15

Corresponding author: E-mail: michelle.christine16@gmail.com (M.C. Chirinos-Arias).




DOI: http://dx.doi.org/10.17268/sci.agropecu.2015.01.02

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