Characterization of Amaranthus cruentus L. seed proteins by 2-DE and LC/MS–MS: Identification and cloning of a novel late embryogenesis-abundant protein

Amaranth is taking great attention as an important cereal crop that could fulfill food requirements for the growing population, especially in developing countries. However, the protein composition of these seeds is not well known yet. We have used the proteomics tools to characterize amaranth seed proteome. About 400 proteins spots were resolved on two-dimensional gel electrophoresis (2-DE) and protein spots were analyzed by LC-MS/MS (liquid chromatography tandem mass spectrometry). Identified proteins were related to stress and defense responses, metabolic, respiratory, and oxide-reduction processes. One abundant spot was identified as a Late Embryogenesis Abundant (LEA) protein and the gene was cloned and characterized. The AcLEA cDNA contains a 418 bp open reading frame (ORF) encoding a polypeptide of 139 amino acids. Bioinformatics analysis showed that AcLEA belongs to LEAs Group 5. Proteomics is a powerful technique that could be used even in non-sequenced organisms such as amaranth. The obtained information reveals that amaranth seed, beyond the classical seed storage proteins, contains proteins related to protection against stress. The identification of these proteins opens the door to the application of new strategies to improve the quality of amaranth production.     

Quantitative proteomic analysis of wheat grain proteins reveals differential effects of silencing of omega-5 gliadin genes in transgenic lines

Novel wheat lines with altered flour compositions can be used to decipher the roles of specific gluten proteins in flour quality. Grain proteins from transgenic wheat lines in which genes encoding the omega-5 gliadins were silenced by RNA interference (RNAi) were analyzed by quantitative 2-dimensional gel electrophoresis (2-DE). The precise effects of the genetic modifications on the proteome were assessed in four homozygous lines generated with the same RNAi construct. Two of the lines showed >80% decreases in omega-5 gliadins with only small changes in the levels of other gluten proteins. In the other two lines, omega-5 gliadins were not detectable by 2-DE. However, there were notable reductions in all omega-1,2 gliadins. Small decreases in several other gluten proteins were also detected in one of the lines. The other line showed notable decreases in three HMW-GS and an s-type LMW-GS, increases in two m-type LMW-GS and several alpha gliadins, as well as increases in both serpins and triticin. The study demonstrates that the same RNAi construct can have differential effects on the wheat grain proteome and highlights the importance of detailed proteomic analyses of transgenic grain prior to selecting lines for further assessment of flour quality and allergenic potential.     

高温逆境对黄金葛叶片中渗透调节物质积累的影响

以3~5个叶片的黄金葛扦插苗为试材,采用单因素随机区组设计,设4个温度处理,3次重复,测定不同温度、不同时间处理下黄金葛叶片内渗透调节物质含量的变化。结果表明,在同一时间高温胁迫下,随温度的不断升高,黄金葛叶片的可溶性蛋白含量整体呈下降趋势,可溶性糖含量呈上升趋势;同一温度处理下,在35℃,不同时间处理可溶性蛋白含量和可溶性糖含量变化不大,在45℃处理40 h,可溶性蛋白含量比胁迫初期降低了35.02%,而可溶性糖含量比胁迫初期增加了97.06%。     

梅花鹿茸可溶性蛋白提取工艺及免疫活性

以梅花鹿茸为原料,采用超声波辅助法提取可溶性蛋白。以鹿茸可溶性蛋白得率为评价指标,通过单因素试验研究提取溶剂浓度、超声提取时间、超声提取功率和液料比对鹿茸可溶性蛋白得率的影响,在单因素试验的基础上进行正交试验,优化提取工艺条件。结果表明：鹿茸可溶性蛋白的最优提取工艺条件为,以pH＝10．00、0．05 mol· L-1的碳酸盐缓冲溶液为提取溶剂,液料比10∶1,超声提取功率200 W,超声提取时间20 min。在此条件下,鹿茸可溶性蛋白得率为36．849 mg· g-1。体内免疫活性试验表明,鹿茸蛋白提取物能够提高免疫功能低下小鼠的单核巨噬细胞吞噬指数、血清溶菌酶质量浓度、血清及肝组织中酸性磷酸酶活力,具有良好的免疫活性。     

Investigation of Aegilopsumbellulatafor stripe rust resistance,heading date,and the contents of iron,zinc, and gluten protein

Aegilops umbellulata (UU) is a wheat wild relative that has potential use in the genetic improvement of wheat.  In this study, 46 Ae. umbellulata accessions were investigated for stripe rust resistance, heading date (HD), and the contents of iron (Fe), zinc (Zn), and seed gluten proteins.  Forty-two of the accessions were classified as resistant to stripe rust, while the other four accessions were classified as susceptible to stripe rust in four environments.  The average HD of Ae. umbellulata was significantly longer than that of three common wheat cultivars (180.9 d vs. 137.0 d), with the exception of PI226500 (138.9 d).  The Ae. umbellulata accessions also showed high variability in Fe (69.74–348.09 mg kg^–1) and Zn (49.83–101.65 mg kg^–1) contents. Three accessions (viz., PI542362, PI542363, and PI554399) showed relatively higher Fe (230.96–348.09 mg kg^–1) and Zn (92.46–101.65 mg kg^–1) contents than the others.  The Fe content of Ae. umbellulata was similar to those of Ae. comosa and Ae. markgrafii but higher than those of Ae. tauschii and common wheat.  Aegilops umbellulata showed a higher Zn content than Ae. tauschii, Ae. comosa, and common wheat, but a lower content than Ae. markgrafii.  Furthermore, Ae. umbellulata had the highest proportion of γ-gliadin among all the species investigated (Ae. umbellulata vs. other species=mean 72.11% vs. 49.37%; range: 55.33–86.99% vs. 29.60–67.91%).  These results demonstrated that Ae. umbellulata exhibits great diversity in the investigated traits, so it can provide a potential gene pool for the genetic improvement of these traits in wheat.     

Proximate composition and functional properties of four cultivars of bambara groundnut ( Voandezeia subterranea)

Proximate composition and selected functional properties of four cultivars of bambara groundnut ( Voandezeia subterranea Thoura) were ascertained. Crude protein ranged from 17.5 to 21.1 percent; crude fat 7.3–8.5 percent; total ash 4–5 percent; crude fiber 1.8–2.0 percent; carbohydrate and moisture content for the different cultivars were 53.0–60.8 percent and 7.5–12.3 percent, respectively. The results of functional property determinations indicated that the bulk density ranged from 0.65 to 0.75 g/ml; water binding capacity 2.1–2.9 g/2g sample; oil binding capacity 0.9–1.6 g/2g sample; emulsifying activity 55.-1–60.0 percent and emulsifying stability 10–12 percent. The results show that bambara groundnut has great potential for incorporation into various human foods where it could provide useful plant proteins.     

Effect of two wheat genotypes and Swedish environment on falling number, amylase activities, and protein concentration and composition

Variation in falling number, amylase activities, protein concentration and composition were investigated in two wheat cultivars grown in Sweden over two seasons, in four locations, with four N fertilizer rates, with and without fungicide treatment. The results showed that;

高温胁迫下糙皮侧耳胞内钙离子对热激蛋白基因表达的调控

以糙皮侧耳(Pleurotus ostreatus)菌株CCMSSC00389为材料,研究高温胁迫(40℃、120min)下钙离子的响应及其对热激蛋白的调控。在高温胁迫条件下,在培养基中添加胞内钙离子螯合剂、胞外钙离子螯合剂、钙调蛋白抑制剂、质膜钙通道抑制剂、三磷酸肌醇受体拮抗剂,利用钙离子特异性荧光探针检测菌丝体胞内钙离子含量,采用荧光定量PCR检测热激蛋白的基因表达量。结果表明,高温胁迫促进胞内钙离子浓度升高,增加Hsp60、Grp78、Hsp90、Hsp104等4种热激蛋白的基因表达量;培养基中添加钙离子螯合剂或抑制剂会降低胞内钙离子浓度,抑制这些热激蛋白的基因表达。研究结果表明高温胁迫会诱导糙皮侧耳胞内钙离子浓度升高,参与信号转导,从而调控热激蛋白的表达来缓解高温胁迫引起的伤害。