We have mapped the expression of the tonoplast intrinsic protein （TIP） gene family members in Arabidopsis seeds by fluorescent protein tagging of their genomic sequences and confocal microscopy. Three isoforms （TIP...We have mapped the expression of the tonoplast intrinsic protein （TIP） gene family members in Arabidopsis seeds by fluorescent protein tagging of their genomic sequences and confocal microscopy. Three isoforms （TIP1;1, TIP2;1, and TIP2;2） have distinct patterns of expression in maternal tissues （outer integument and placento-chalazal region）. Two isoforms, TIP3;1 and the previously uncharacterized TIP3;2, are the only detectable TIPs in embryos during seed maturation and the early stages of seed germination. Throughout these developmental stages, both isoforms co-locate to the tono- plast of the protein storage vacuoles, but also appear to label the plasma membrane. Plasma membrane labeling is specific to TIP3;1 and TIP3;2, is independent of the position of the fluorescent protein tag, and appears to be specific to early seed maturation and early germination stages. We discuss these results in the context of the predicted distribution of aquaporins in Arabidopsis seeds.展开更多
The endopeptidases （EPs） in wheat endosperm during seed germination and subsequent seedling growth were characterized by gradient-polyacrylamide gel electrophoresis with gelatin copolymerized into the gel. Four cyst...The endopeptidases （EPs） in wheat endosperm during seed germination and subsequent seedling growth were characterized by gradient-polyacrylamide gel electrophoresis with gelatin copolymerized into the gel. Four cysteine EPs （EP1, EP2, EP3 and EP4） were detected in wheat endosperm during the 7 d growth after seed imbibition. The results also showed that the activities of all of these EPs increased continuously, and EP2 first appeared and had the highest proteolytic activity among the four EPs in this experimental process. The optimum pH and temperature of all four EPs were 4.0 and 40.0 ~C. All EPs were completely inhibited by 25 μmol/L E-64 and had no good thermal stabilities, especially EP1. In addition, these EPs had different substrate specificities to albumins, globulins, gliadins and glutenins; the main storage proteins of mature wheat endosperm. Among them, EP2 had the highest proteolytic activities on globulins, gliadins and glutenins, and might be the most important and specific EP with potential to be tightly correlated with seedling development.展开更多
文摘We have mapped the expression of the tonoplast intrinsic protein （TIP） gene family members in Arabidopsis seeds by fluorescent protein tagging of their genomic sequences and confocal microscopy. Three isoforms （TIP1;1, TIP2;1, and TIP2;2） have distinct patterns of expression in maternal tissues （outer integument and placento-chalazal region）. Two isoforms, TIP3;1 and the previously uncharacterized TIP3;2, are the only detectable TIPs in embryos during seed maturation and the early stages of seed germination. Throughout these developmental stages, both isoforms co-locate to the tono- plast of the protein storage vacuoles, but also appear to label the plasma membrane. Plasma membrane labeling is specific to TIP3;1 and TIP3;2, is independent of the position of the fluorescent protein tag, and appears to be specific to early seed maturation and early germination stages. We discuss these results in the context of the predicted distribution of aquaporins in Arabidopsis seeds.
基金Supported by the National Natural Science Foundation of China (30370851).
文摘The endopeptidases （EPs） in wheat endosperm during seed germination and subsequent seedling growth were characterized by gradient-polyacrylamide gel electrophoresis with gelatin copolymerized into the gel. Four cysteine EPs （EP1, EP2, EP3 and EP4） were detected in wheat endosperm during the 7 d growth after seed imbibition. The results also showed that the activities of all of these EPs increased continuously, and EP2 first appeared and had the highest proteolytic activity among the four EPs in this experimental process. The optimum pH and temperature of all four EPs were 4.0 and 40.0 ~C. All EPs were completely inhibited by 25 μmol/L E-64 and had no good thermal stabilities, especially EP1. In addition, these EPs had different substrate specificities to albumins, globulins, gliadins and glutenins; the main storage proteins of mature wheat endosperm. Among them, EP2 had the highest proteolytic activities on globulins, gliadins and glutenins, and might be the most important and specific EP with potential to be tightly correlated with seedling development.
