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Salt-induced delay in cotyledonary globulin mobilization is abolished by induction of proteases and leaf growth sink strength at late seedling establishment in cashew
Authors:Luiz Ferreira Aguiar Ponte,André   Luis Coelho da Silva,Fabrí  cio Eulá  lio Leite Carvalho,Josemir Moura Maia,Eduardo Luiz Voigt,Joaquim Albenisio Gomes Silveira
Affiliation:1. Centro de Ciências Agrárias e Biológicas, Universidade Estadual Vale do Acaraú, CEP 62040-370, Sobral, CE, Brazil;2. Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Ceará, CEP 60451-970, Fortaleza, Ceará, Brazil;3. Campos IV – Catolé do Rocha, Centro de Ciências Humanas e Agrárias, CCHA, Universidade Estadual da Paraiba, Paraiba, Brazil;4. Laboratório de Estudos em Biotecnologia Vegetal, Departamento de Biologia Celular e Genética, Universidade Federal do Rio Grande do Norte, RN, Brazil
Abstract:Seedling establishment in saline conditions is crucial for plant survival and productivity. This study was performed to elucidate the biochemical and physiological mechanisms involved with the recovery and establishment of cashew seedlings subjected to salinity. The changes in the Na+ levels and K/Na ratios, associated with relative water content, indicated that osmotic effects were more important than salt toxicity in the inhibition of seedling growth and cotyledonary protein mobilization. Salinity (50 mM NaCl) induced a strong delay in protein breakdown and amino acid accumulation in cotyledons, and this effect was closely related to azocaseinolytic and protease activities. In parallel, proline and free amino acids accumulated in the leaves whereas the protein content decreased. Assays with specific inhibitors indicated that the most important proteases in cotyledons were of serine, cysteine and aspartic types. Proteomic analysis revealed that most of the cashew reserve proteins are 11S globulin-type and that these proteins were similarly degraded under salinity. In the late establishment phase, the salt-treated seedlings displayed an unexpected recovery in terms of leaf growth and N mobilization from cotyledon to leaves. This recovery coordinately involved a great leaf expansion, decreased amino acid content and increased protein synthesis in leaves. This response occurred in parallel with a prominent induction in the cotyledon proteolytic activity. Altogether, these data suggest that a source–sink mechanism involving leaf growth and protein synthesis may have acted as an important sink for reserve mobilization contributing to the seedling establishment under salinity. The amino acids that accumulated in the leaves may have exerted negative feedback to act as a signal for the induction of protease activity in the cotyledon. Overall, these mechanisms employed by cashew seedlings may be part of an adaptive process for the efficient rescue of cotyledonary proteins, as the cashew species originates from an environment with N-poor soil and high salinity.
Keywords:2-DE, two dimensional electrophoresis   DAS, days after sowing   E-64, 10   μM trans-epoxysuccinyl-l-leucylamide (4-guanidine) butane   ESI, electrospray ionization   PEPS, pepstatin   PMSF, phenylmethylsulfonyl fluoride   Q-TOF, quadrupole time-of-flight   RWC, relative water content   SSP, seed storage protein   UA, unity of activity   Ψs, osmotic potential
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