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Cloning and expression of a sorghum gene with homology to maize vp1. Its potential involvement in pre-harvest sprouting resistance
Authors:Carrari  Fernando  Perez-Flores  Laura  Lijavetzky  Diego  Enciso  Silvina  Sanchez  Rodolfo  Benech-Arnold  Roberto  Iusem  Norberto
Affiliation:(1) Laboratorio de Fisiología y Biología Molecular, FCEyN, Universidad de Buenos Aires, Ciudad Universitaria, Argentina;(2) Departamento de Ciencias de la Salud, DCBS, Universidad Autónoma Metropolitana-Iztapalapa, D.F. México;(3) CICV, INTA, Instituto de Biotecnología, Castelar, Argentina;(4) IFEVA, Facultad de Agronomía, Universidad de Buenos Aires, Argentina
Abstract:Pre-harvest sprouting (PHS) in sorghum is related to the lack of a normal dormancy level during seed development and maturation. Based on previous evidence that seed dormancy in maize is controlled by the vp1 gene, we used a PCR-based approach to isolate two Sorghum bicolor genomic and cDNA clones from two genotypes exhibiting different PHS behaviour and sensitivity to abscisic acid (ABA). The two 699 amino acid predicted protein sequences differ in two residues at positions 341 (Gly or Cys within the repression domain) and 448 (Pro or Ser) and show over 80, 70 and 60% homology to maize, rice and oat VP1 proteins respectively.Expression analysis of the sorghum vp1 gene in the two lines shows a slightly higher level of vp1 mRNA in the embryos susceptible to PHS than in those resistant to PHS during embryogenesis. However, timing of expression was different between these genotypes during this developmental process. Whereas for the former the main peak of expression was observed at 20 days after pollination (DAP), the peak in the latter was found at later developmental stages when seed maturation was almost complete.Under favourable germination conditions and in the presence of fluridone (an inhibitor of ABA biosynthesis), sorghum vp1 mRNA showed to be consistently correlated with sensitivity to ABA but not with ABA content and dormancy.
Keywords:dormancy  germination  pre-harvest sprouting  Sorghum bicolor  vp1 gene.
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