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Sensitivity of Seeds to Chemical Mutagens, Detection of DNA Polymorphisms and Agro-Metrical Traits in M1 Generation of Coffee (Coffea arabica L.)

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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.

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Acknowledgments

This study was financed by the University of Costa Rica, the Ministerio de Ciencia, Tecnology Telecomunicaciones (MICITT) and the Consejo Nacional para Investigaciones Cientcas y Tecnolas (CONICIT) (project No. 111-B5- 140; FI-030B-14). The authors would like to thank Dr. Paul Hanson (School of Biology, University of Costa Rica) for language correction of the manuscript.

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  1. Laboratorio Biotecnología de Plantas, Escuela de Biología, Universidad de Costa Rica, 2060, San Pedro, Costa Rica

    César Vargas-Segura, Emmanuel López-Gamboa, Marta Valdez-Melara & Andrés Gatica-Arias

  2. Centro Nacional de Innovaciones Biotecnológicas (CENIBiot), CeNAT-CONARE, 1174-1200, San José, Costa Rica

    Emanuel Araya-Valverde

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  1. César Vargas-Segura
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  2. Emmanuel López-Gamboa
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  3. Emanuel Araya-Valverde
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  4. Marta Valdez-Melara
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Contributions

C.V.S designed and performed the experiments and analyzed data; E.L.G performed the phenotyping of the M1 mutants; E.A.V. performed AFLPs and analyzed data; M.V.M. discussed the results and edited the paper; A.G.A conceived the project, designed and coordinated the experiments, analyzed data, and wrote the paper.

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Correspondence to Andrés Gatica-Arias.

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The authors declare that they have no conflict of interest. All authors read and approved the final manuscript.

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Vargas-Segura, C., López-Gamboa, E., Araya-Valverde, E. et al. Sensitivity of Seeds to Chemical Mutagens, Detection of DNA Polymorphisms and Agro-Metrical Traits in M1 Generation of Coffee (Coffea arabica L.). J. Crop Sci. Biotechnol. 22, 451–464 (2019). https://doi.org/10.1007/s12892-019-0175-0

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