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Seeds store lipids in the form of lipid bodies (LBs) for germination and early seedling growth. LBs can be easily isolated by the established floating-extraction method from oleaginous seeds containing a large quantity of LBs. Compared to oleaginous seeds, maize and other cereal seeds contain a small quantity of LBs, so it is difficult to isolate a sufficient quantity of LBs from their embryos for 2DE-based proteomic analysis. At present, only a limited number of LBs-associated proteins in maize embryos have been identified. We here reported a modified floating-extraction method using polyvinylidene difluoride disc to collect floating LBs from maize embryo extracts. The LBs-associated proteins were resolved with two-dimensional electrophoresis and identified with mass spectrometry. As a result, several well-known LBs proteins were identified in the purified LBs fraction, such as oleosin, caleosin, and steroleosin. We also identified another two LBs proteins, corticosteroid 11-β-dehydrogenase 1 and 11-β-hydroxysteroid dehydrogenase-like 5. In particular, steroleosin, corticosteroid 11-β-dehydrogenase 1, 11-β-hydroxysteroid dehydrogenase-like, and hydroxyproline-rich glycoprotein were found as the most abundant protein components in maize LBs. The data set of maize LBs subproteome would provide insights into functional research of LBs-associated proteins during seed development and germination. Additionally, the protocol developed here is expected to be applicable for isolating LBs in other seeds or tissues containing a low quantity of LBs.
Flaveria bidentis, an annual grass native to South America, has invaded into many countries all over the world, including South Africa and Egypt in Africa, Britain and France in Europe, Japan in Asia, and Australia. In China, this plant species has been widely distributed in Hebei province since 2001 of its first discovery in Tianjin. In salinized soil of northern China, F. bidentis has formed mono dominant communities owing to its opportunistic characteristics. In this study, we investigated germination rate, germination energy, germination index and vigor index in response to different saline-alkaline stresses. Lengths of seedling, hypocotyl, and radicle were also examined. During germination process, germination rate, germination energy, germination index, and vigor index decreased due to higher salinity and alkalinity (pH). Hypocotyl elongation was stimulated at low salinity and alkalinity, but decreased with increasing salinity and alkalinity. The lengths of radicle and seedling were inhibited sharply with increasing salinity and alkalinity. These results suggest that a reciprocal enhancement between salt stress and alkaline stress was a characteristic feature during seed germination of F. bidentis. Ungerminated seeds resumed to germinate after transferred to distilled water, indicating that seeds remained viable during their exposure to saline-alkaline stresses. Stepwise regression analysis indicated that the effects of salinity, alkalinity (pH), and buffer capacity on seed germination and seedling growth were significantly different in magnitude. Salinity and alkalinity (pH) were the dominant factors in seed germination and seedling growth of F. bidentis respectively. burther, the results of this study suggest that F. bidentis has developed excellent adaptative strategy in its early stage of life cycle which partially explains its current invasion success in northern China.
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