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Universidad Autónoma de Nuevo León (2014)

Identificación de marcadores moleculares ligados a caracteres de interés en materiales élite de sorgo dulce Sorghum bicolor (L.) Moench de la FA-UANL.

Cerda Hurtado, Ivon Montserrat

Titre : Identificación de marcadores moleculares ligados a caracteres de interés en materiales élite de sorgo dulce Sorghum bicolor (L.) Moench de la FA-UANL.

Auteur : Cerda Hurtado, Ivon Montserrat

Université de soutenance : Universidad Autónoma de Nuevo León

Grade : Maestría en Ciencias en Producción Agrícola) UANL, 2014.

Résumé
The use of molecular markers in breeding programs has proven to be an effective tool in selection procedures. AFLP and RAPD markers could be effective for identification of polymorphisms and the construction of grouping by genetic distances. SCAR development allows for reproducible results of PCR-based polymorphic RAPD markers therefore be used in identifying high sorghum genotypes in the content of sugars in the juice from the stem.
The objective of the study was to develop a reliable and reproducible molecular marker that can be used in selection programs focused on sweet sorghum bioethanol production.
Fourteen sorghum genotypes were used : FA-UANL 37A, FA-UANL 33A, FAUANL 35A, AN601, FA-UANL 39, FA-UANL 5, KANSAS COLLIER, FA-UANL 33A x KANSAS COLLIER, AB601 x FA-UANL 39, FA-UANL 35A x FA-UANL 39, FAUANL 33A x FA-UANL 5 and FA-UANL 37A x FA-UANL5 and FORTUNA. Duplex reactions for 30 primer combinations EcoRI/MseI for AFLP markers were conducted, along with the test of 180 RAPD primers. 53 RAPD fragments present in the genotypes higher in sugars and absent at lower sugar genotypes were identified, purified, cloned and sequenced.
There were obtained 549 AFLP and 256 RAPD fragments. Two pairs of SCAR primers were designed : the set of primers 4 amplified a band of 231 bp present in the high sugar genotypes (A601 x FA-UANL 39, FA-UANL 35A x FA-UANL 39, FAUANL 33A x FA-UANL 5 and FA-UANL 37A x FA-UANL 5). This band did not amplified in any other genotype, but neither was present in any parent ; the set of primers 7 amplified a band of 341 bp present in the high sugar genotypes (A601 x FA-UANL 39 and FA-UANL 33A x FA-UANL 5), this band was not amplified in any other genotype, but was found in the B lines of the parents (FA-UANL 39 and FAUANL 5) and it was also present in the parent KANSAS COLLIER.
The AFLP and RAPD markers were efficient for estimating genetic variation in sorghum by presenting a genetic distance coefficient from 0.42 (Dice √ (S-1)) in the cluster tools. AFLP technique proved to be advantageous to evaluate the variation between genotypes, having a higher yield of bands by primers set ; however, they are not suitable for designing SCAR primers by size. Using RAPD markers allowed the design of two SCAR primer sets from fragments that aligned with sorghum reported genomic sequences at NCBI.

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