Selección de mutantes de arroz (Oryza sativa) con tolerancia a salinidad obtenidos por radiación gamma
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Fecha
2020
Autores
Fernández Acuña, Alexis Ángel
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Universidad Nacional, Costa Rica
Resumen
Esta investigación tuvo como objetivo aprovechar la variabilidad genética inducida por la radiación gamma cobalto 60 (Co60) en el cultivo del arroz, para seleccionar mutantes con tolerancia a la salinidad. Se utilizó 1,5 kg semillas de arroz de la variedad CR5272 irradiadas a una dosis de 400 Gy y se multiplicó para obtener la generación M2 en condiciones de campo. Se valoró la susceptibilidad a la salinidad (NaCl) de esta variedad mediante la determinación de la dosis letal media (DL50) usando valores de conductividad eléctrica (CE) de 2 a 20 dS/m (deciSiemens por metro) con incrementos de dos unidades entre cada valor. Para ello se utilizó el cultivo hidropónico, utilizando la solución nutritiva descrita por Yoshida et al. (1976), para el crecimiento de plantas de arroz. En campo se logró cosechar 14 Kg de semilla M2. En invernadero se determinó que la DL50 del factor de estrés salino (factor de selección) corresponde a 11.75 dS/m. No obstante, se usó 14 dS/m como factor de estrés debido a que esta dosis fue más eficiente para discriminar mutantes tolerantes de los susceptibles al compararse con la de 11.75 dS/m. Una vez obtenida la semilla M2 (14 kg) en campo y definido el nivel de estrés salino a ser aplicado a la población segregante se tomaron 9450 semillas M2 y se sembraron en invernadero en el sistema hidropónico, al cual se le adicionó la solución nutritiva ya descrita salinizada a 14 dS/m de CE desde la germinación hasta los 21 días de edad. En esta primera fase se seleccionaron 30 mutantes, los cuales fueron trasplantados a un medio sin sal usando el mismo sistema hidropónico para que continuaran su crecimiento hasta la etapa de prefloración. En la cual fueron sometidas nuevamente al factor de estrés durante 25 días. En esta segunda evaluación sobrevivieron ocho mutantes promisorios de los cuales se recuperó semilla para ser incrementada. A lo largo de todo el ciclo fenológico del cultivo se evaluaron diferentes variables agronómicas con el fin de describir a los mutantes promisorios. Esta investigación demostró que la técnica de inducción de mutaciones usando irradiaciones gamma Co60 es capaz de generar mutantes promisorios con tolerancia a salinidad, los cuales pueden ser usados en la obtención de nuevas variedades que contribuyan a enfrentar el avance de la salinización de los suelos de Costa Rica.
The objective of this research was to take advantage of the genetic variability induced by cobalt-60 (Co60) gamma irradiation in rice to select mutants with salinity tolerance. We used 1.5 kg of rice seeds of variety CR5272 irradiated at a dose of 400 Gy and multiplied to obtain the M2 generation under field conditions. The susceptibility to salinity (NaCl) of this variety was assessed by determining the mean lethal dose (LD50) using electrical conductivity (EC) values from 2 to 20 dS/m (deciSiemens per meter) with increments of two units between each value. Hydroponic culture was used for this purpose, using the nutrient solution described by Yoshida et al. (1976), for the growth of rice plants. In the field, 14 kg of M2 seed were harvested. In the greenhouse, it was determined that the LD50 of the salt stress factor (selection factor) corresponds to 11.75 dS/m. However, 14 dS/m was used as the stress factor because this dose was more efficient in discriminating tolerant mutants from susceptible ones when compared to 11.75 dS/m. Once the M2 seed (14 kg) was obtained in the field and the level of salt stress to be applied to the segregating population was defined, 9450 M2 seeds were taken and sown in the greenhouse in the hydroponic system, to which the nutrient solution already described was added, salinized at 14 dS/m of EC from germination until 21 days of age. In this first phase, 30 mutants were selected and transplanted to a medium without salt using the same hydroponic system to continue their growth until the pre-flowering stage. In which they were again subjected to the stress factor for 25 days. In this second evaluation, eight promising mutants survived and seed was recovered from them to be increased. Throughout the phenological cycle of the crop, different agronomic variables were evaluated in order to describe the promising mutants. This research demonstrated that the mutation induction technique using Co60 gamma irradiation is capable of generating promising mutants with tolerance to salinity, which can be used in the breeding of new varieties that contribute to face the advance of salinization in Costa Rican soils.
The objective of this research was to take advantage of the genetic variability induced by cobalt-60 (Co60) gamma irradiation in rice to select mutants with salinity tolerance. We used 1.5 kg of rice seeds of variety CR5272 irradiated at a dose of 400 Gy and multiplied to obtain the M2 generation under field conditions. The susceptibility to salinity (NaCl) of this variety was assessed by determining the mean lethal dose (LD50) using electrical conductivity (EC) values from 2 to 20 dS/m (deciSiemens per meter) with increments of two units between each value. Hydroponic culture was used for this purpose, using the nutrient solution described by Yoshida et al. (1976), for the growth of rice plants. In the field, 14 kg of M2 seed were harvested. In the greenhouse, it was determined that the LD50 of the salt stress factor (selection factor) corresponds to 11.75 dS/m. However, 14 dS/m was used as the stress factor because this dose was more efficient in discriminating tolerant mutants from susceptible ones when compared to 11.75 dS/m. Once the M2 seed (14 kg) was obtained in the field and the level of salt stress to be applied to the segregating population was defined, 9450 M2 seeds were taken and sown in the greenhouse in the hydroponic system, to which the nutrient solution already described was added, salinized at 14 dS/m of EC from germination until 21 days of age. In this first phase, 30 mutants were selected and transplanted to a medium without salt using the same hydroponic system to continue their growth until the pre-flowering stage. In which they were again subjected to the stress factor for 25 days. In this second evaluation, eight promising mutants survived and seed was recovered from them to be increased. Throughout the phenological cycle of the crop, different agronomic variables were evaluated in order to describe the promising mutants. This research demonstrated that the mutation induction technique using Co60 gamma irradiation is capable of generating promising mutants with tolerance to salinity, which can be used in the breeding of new varieties that contribute to face the advance of salinization in Costa Rican soils.
Descripción
Licenciatura en Ingeniería Agronómica con modalidad de graduación: tesis
Palabras clave
ARROZ, RICE, ENERGÍA RADIANTE, RADIANT ENERGY, SALINIDAD, SALINITY, MUTACIONES, MUTATIONS, GENÉTICA VEGETAL, PLANT GENETICS