[Establish quality evaluation method based on standard decoction of Danshen extract].

The quality of Danshen extract granules on market is largely different from each other mainly due to the heterogeneous quality of raw materials of Salvia miltiorrhiza, various producing procedures and lack of good quality evaluation method. Formula granule and "standard decoction" have the same quality. In this paper, a systematic evaluation method for the quality of Danshen decoction was established from the perspective of "standard decoction", in order to explore the main factors affecting the quality uniformity of Danshen extract granules. Danshen standard decoction was prepared; then the fingerprint method was developed to determine the content of salvianolic acid B; and the main peaks in the fingerprint were identified with UPLC-QTOF/MS to clarify the chemical compositions of Danshen decoction. Three indexes were calculated to evaluate the stability of whole process, including the extraction ratio; transfer rate of index components and pH value. The results showed that the main components of Danshen decoction were phenolic acids, while the extraction rate, the transfer rate of salvianolic acid B and pH value were in a relatively stable level, and the similarity in the fingerprint of standard decoction was high, indicating that the preparation procedure was stable. The level of salvianolic acid B in the standard decoction was in a large range, which was mainly due to the difference in the quality of Salviae Miltiorrhizae Radix et Rhizoma.

Proteomics insight into the biological safety of transgenic modification of rice as compared with conventional genetic breeding and spontaneous genotypic variation.

The potential of unintended effects caused by transgenic events is a key issue in the commercialization of genetically modified (GM) crops. To investigate whether transgenic events cause unintended effects, we used comparative proteomics approaches to evaluate proteome differences in seeds from 2 sets of GM indica rice, herbicide-resistant Bar68-1 carrying bar and insect-resistant 2036-1a carrying cry1Ac/sck, and their respective controls D68 and MH86, as well as indica variety MH63, a parental line for breeding MH86, and japonica variety ZH10. This experimental design allowed for comparing proteome difference caused by transgenes, conventional genetic breeding, and natural genetic variation. Proteomics analysis revealed the maximum numbers of differentially expressed proteins between indica and japonica cultivars, second among indica varieties with relative small difference between MH86 and MH63, and the minimum between GM rice and respective control, thus indicating GM events do not substantially alter proteome profiles as compared with conventional genetic breeding and natural genetic variation. Mass spectrometry analysis revealed 234 proteins differentially expressed in the 6 materials, and these proteins were involved in different cellular and metabolic processes with a prominent skew toward metabolism (31.2%), protein synthesis and destination (25.2%), and defense response (22.4%). In these seed proteomes, proteins implicated in the 3 prominent biological processes showed significantly different composite expression patterns and were major factors differentiating japonica and indica cultivars, as well as indica varieties. Thus, metabolism, protein synthesis and destination, and defense response in seeds are important in differentiating rice cultivars and varieties.

Comparative proteomic study reveals the involvement of diurnal cycle in cell division, enlargement, and starch accumulation in developing endosperm of Oryza sativa.

The development and starch accumulation of cereal endosperms rely on the sugar supply of leaves, which is subject to diurnal cycles, and the endosperm itself also experiences a light/dark switch. However, revealing how the cereal endosperm responds to diurnal input remains a major challenge. We used comparative proteomic approaches to probe diurnally affected processes in rice endosperm (Oryza sativa) 10 days after flowering under 12-h light/12-h dark. Starch granules in rice endosperm showed a growth ring structure under a normal light/dark cycle but not under constant light. Sucrose showed a high level in light and low level in dark. Two-dimensional (2-D) differential in-gel electrophoresis-based proteomic analysis revealed 101 protein spots diurnally changed and 91 identities, which were involved in diverse processes with preferred distribution in stress response, protein synthesis/destination and metabolism. Proteins involved in cell division showed high expression in light and those in cell enlargement and cell wall synthesis high in dark, while starch synthesis proteins were light-downregulated and dark-upregulated. Redox homeostasis-associated proteins showed in-phase peaks under light and dark. These data demonstrate diurnal input-regulated diverse cellular and metabolic processes in rice endosperm, and coordination among these processes is essential for development and starch accumulation with diurnal input.

Integrated proteomic and cytological study of rice endosperms at the storage phase.

The endosperm at the storage phase undergoes a series of coordinated cellular and metabolic events, including starchy endosperm cell death, starch synthesis, and starch granule packaging, which leads to efficient accumulation of starch. However, the mechanism underlying the interconnections remains unknown. We used integrated proteomic and cytological approaches to probe the interconnections in rice (Oryza sativa) endosperm at the storage phase from 12 to 18 days after flowering (DAF). Starch granule packaging was completed first in the inner part of endosperm at 15 DAF and spread to almost the entire endosperm at 18 DAF. Programmed starchy endosperm cell death occurred after the starch granule packaging. Endogenous H(2)O(2) was detectable in the inner part of endosperm at 12 DAF and the region beyond the inner part at 15 DAF, with an H(2)O(2) burst at 15 DAF. Proteomics analysis with 2-D fluorescent difference gel electrophoresis and matrix-assisted laser-desorption ionization time-of-flight/time-of-flight mass spectrometry revealed 317 proteins, including almost all known antioxidants, differentially expressed throughout the 3 stages of the developmental phase. More than two-thirds of the 317 proteins were potential thioredoxin targets, with a preferential skew toward central carbon metabolism, alcoholic fermentation, starch metabolism, amino acid metabolism, and protein synthesis or folding. These proteins implicated in starch synthesis and gluconeogenesis were upregulated, whereas those involved in anabolism of biomacromolecules such as proteins, lipids, and cell wall components were downregulated, with upregulated expression of proteins involved in catabolism of these biomacromolecules, which suggests remobilization of nutrients for starch synthesis. These data suggested important roles of the H(2)O(2)-antioxidant interface in coordinating starch accumulation, programmed cell death of starchy endosperm, and remobilization of nutrients during the cell death.