Hydrolysis of lipid and protein reserves in loblolly pine seeds in relation to protein electrophoretic patterns following imbibition

Abstract

The correlation between changes in seed protein electrophoretic patterns and the hydrolysis of lipid and protein reserves of loblolly pine (Pinus taeda L.) seed was studied. Seeds were incubated at 30°C for up to 12 days following stratification, then megagametophytes and embryos were assayed for lipid and protein content after each day of imbibition. The megagametophyte of mature seed was found to contain 20% lipid and 12% storage protein on a fresh weight basis. The embryo contained 26% lipid and 15% protein. Both lipid and protein reserves were depleted constantly following imbibition. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of soluble and insoluble protein fractions showed a 60 kDa protein that was representative of crystalloid‐like proteins. These crystalloid‐like proteins comprised 85% of the insoluble protein storage reserves. A small number of insoluble storage proteins, including a 47 kDa protein, were distinct in that they were unaffected by 2‐mercaptoethanol treatment. The soluble fractions from both tissues were labelled with [35S]‐methionine, and incorporation was visualized by two‐dimensional electrophoresis. Proteins were found to belong to one of three categories, those synthesized constitutively (comprising the bulk of newly synthesized proteins), those synthesized during germination or those synthesized after radicle emergence. Accompanying seed reserve hydrolysis were developmental shifts in protein pattern and synthesis, suggesting the possibility that control of hydrolysis is at the level of enzyme accumulation.