Sensitivity of Seeds to Chemical Mutagens, Detection of DNA Polymorphisms and Agro-Metrical Traits in M1 Generation of C
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J. Crop Sci. Biotech. 2019 (December) 22 (5) : 451 ~ 464 DOI No. 10.1007/s12892-019-0175-0 RESEARCH ARTICLE
Sensitivity of Seeds to Chemical Mutagens, Detection of DNA Polymorphisms and Agro-Metrical Traits in M1 Generation of Coffee (Coffea arabica L.) César Vargas-Segura1, Emmanuel López-Gamboa1, Emanuel Araya-Valverde2, Marta Valdez-Melara1, Andrés Gatica-Arias1* 1
Laboratorio Biotecnología de Plantas, Escuela de Biología, Universidad de Costa Rica, 2060 San Pedro, Costa Rica Centro Nacional de Innovaciones Biotecnológicas (CENIBiot), CeNAT-CONARE, 1174-1200, San José, Costa Rica
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Received: June 25, 2019 / Revised: June 26, 2019 / Accepted: July 01, 2019 Ⓒ Korean Society of Crop Science and Springer 2019
Abstract Coffee (Coffea arabica L.) is threatened by biotic and abiotic stresses. Nevertheless, the breeding of Arabica coffee is restricted due to its low genetic diversity. Crop improvement via mutagenesis represents an alternative for increasing genetic variability and facilitating breeding. In this sense, coffee seeds cv. Catuaí were treated for 8 h with a solution of sodium azide (NaN3) (0, 50, 75, 100, and 125 mM) and ethyl methane sulfonate (EMS) (0, 80, 160, 240, 320, and 400 mM). The genetic variability induced in coffee plants after mutagenic treatment with sodium azide was determined by RAPD and AFLP analyses. As the concentration of applied NaN3 and EMS increased, the germination, seedling height, and root length decreased. The LD50 values for NaN3 and EMS were between 50-75 mM and 160-240 mM, respectively. For the 12 RAPD primers evaluated, a total of 46 fragments were obtained of which 34 were polymorphic bands (74%). The amplification with six AFLP selective primer combinations allowed the identification of 36 polymorphisms (17.8%). The analysis revealed that both NaN3 and EMS induced variability within the DNA regions amplified with AFLP and RAPD markers. Finally, under field conditions, significant differences were noticed with respect to plant height, number of nodes in the orthotropic stem, and number of branches of the M1 mutant (NaN3-treated) plants compared to the non-mutant plants. Optimal conditions for NaN3 and EMS mutagenesis using seeds were determined and the optimized conditions have been used to generate a NaN3 mutant M1 coffee var. Catuaí population. Key words : Coffee, chemical mutagenesis, EMS, sodium azide, AFLP, RAPD, M1 generation
Introduction The flavor, aroma, and the stimulating effects of caffeine have converted coffee into one of the most highly valuable non-alcoholic beverages for consumption worldwide (Ivamoto et al. 2017). C. arabica L. (2n=4x=44) and C. canephora Pierre ex A. Froehner (2n=2x=22) are responsible for 70 and 30%, respectively, of coffee production (Campos et al. 2017). Nevertheless, several biotic (diseases and pests) and abiotic (drought, heat stress, and salinity) factors threaten coffee production (dos Santos et al. 2011; Silva et al. 2006). For this reason, Arabica coffee cultivars have been conventionally bred to confer resistance against bio
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