Proteome and phosphoproteome characterization reveals new response and defense mechanisms of Brachypodium distachyon leaves under salt stress.

Salinity is a major abiotic stress affecting plant growth and development. Understanding the molecular mechanisms of salt response and defense in plants will help in efforts to improve the salt tolerance of crops. Brachypodium distachyon is a new model plant for wheat, barley, and several potential biofuel grasses. In the current study, proteome and phosphoproteome changes induced by salt stress were the focus. The Bd21 leaves were initially treated with salt in concentrations ranging from 80 to 320 mm and then underwent a recovery process prior to proteome analysis. A total of 80 differentially expressed protein spots corresponding to 60 unique proteins were identified. The sample treated with a median salt level of 240 mm and the control were selected for phosphopeptide purification using TiO2 microcolumns and LC-MS/MS for phosphoproteome analysis to identify the phosphorylation sites and phosphoproteins. A total of 1509 phosphoproteins and 2839 phosphorylation sites were identified. Among them, 468 phosphoproteins containing 496 phosphorylation sites demonstrated significant changes at the phosphorylation level. Nine phosphorylation motifs were extracted from the 496 phosphorylation sites. Of the 60 unique differentially expressed proteins, 14 were also identified as phosphoproteins. Many proteins and phosphoproteins, as well as potential signal pathways associated with salt response and defense, were found, including three 14-3-3s (GF14A, GF14B, and 14-3-3A) for signal transduction and several ABA signal-associated proteins such as ABF2, TRAB1, and SAPK8. Finally, a schematic salt response and defense mechanism in B. distachyon was proposed.

A recessive gene controlling male sterility sensitive to short daylength/low temperature in wheat (Triticum aestivum L.).

Utilization of a two-line breeding system via photoperiod-thermo sensitive male sterility has a great potential for hybrid production in wheat (Triticum aestivum L.). 337S is a novel wheat male sterile line sensitive to both short daylength/low temperature and long daylength/high temperature. Five F(2) populations derived from the crosses between 337S and five common wheat varieties were developed for genetic analysis. All F(1)'s were highly fertile while segregation occurred in the F(2) populations with a ratio of 3 fertile:1 sterile under short daylength/low temperature. It is shown that male sterility in 337S was controlled by a single recessive gene, temporarily designated as wptms3. Bulked segregant analysis (BSA) coupled with simple sequence repeat (SSR) markers was applied to map the sterile gene using one mapping population. The wptms3 gene was mapped to chromosome arm 1BS and flanked by Xgwm413 and Xgwm182 at a genetic distance of 3.2 and 23.5 cM, respectively. The accuracy and efficiency of marker-assisted selection were evaluated and proved essential for identifying homozygous recessive male sterile genotypes of the wptms3 gene in F(2) generation.

[Simulation model of barley leaf area index].

To simulate leaf area index (LAI) accurately is the key for the prediction of crop growth and yield in a crop growth model. Based on the analysis of the dynamic changes in the LAI of high-yielding barley cultivars in Wuhan and Yangzhou, a simulation model of barley LAI was established, in which, the LAI was the function of expansion coefficient of LAI for cultivar genetic property, climatic factors such as daily air temperature difference, sunshine hours, and accumulation of photosynthetic available radiation after sowing (sigma PAR), and limitation indices of water and nutrients. It was indicated that the maximum LAI and optimal LAI at the stages of booting and heading were not the same conception, but differed significantly. The model was tested by the field experiments with different barley cultivars under different sowing dates and nitrogen application rates in Yangzhou, Nanjing, and Kunming. The results showed that this model gave the good predictions of LAI at different development stages, with the RMSE values ranged in 0.742 and 2.865, and averaged 1.348. The simulated and observed LAI values were significantly positively correlated, and the correlation coefficient from y = x regression analysis was between 0.511 and 0.954.

[Wheat transformation by electroporation with ring electrode].

Electroporation has been used effectively to deliver DNA into the tissue of intact wheat immature embryos. The physical parameters of electroporation were 770 V/cm field strength, 800 microF capacitor and 100 microg/mL of plasmid DNA, containing bar and GUS gene. The electroporation was carried out by a portable and permanent ring electrode that can be fitted into the wells of 24-well plates. The samples were pulsed three times. Integration of the introduced genes into the genome of transgenic wheat plants was shown by PCR and Southern blot analysis. Transformation frequency was 7.5%, that is higher than that of the 4.2% frequency by microprojectile bombardment.

[Obtainment of transgenic wheat with the insecticidal lectin from snowdrop (Galanthus nivalis agglutinin; GNA) gene and analysis of resistance to aphid].

Snowdrop lectin (Galanthus nivalis agglutinin; GNA) is toxic to sap sucking injurious insects of Homopteran. A new gna gene has been transferred into common spring wheat Zhong60634 and winter wheat Yumai66 with high yield by using the biolistic transformation method. Transgenic wheat plants have been obtained in both of the two varieties. Two transgenic plants (T0) have been obtained from the bombarded 535 immature embryos of Zhong60634. Bioassay results show that the development of aphid could be slowed down and the survival rate of young aphid could be reduced by gna gene. Seventeen transgenic plants (T0) were obtained from the bombarded 4636 immature embryos of Yumai66. Twenty plantlets with good resistance to Rhopalosiphum padi and Macrosiphum avenae, which are mainly aphid in north wheat area, were identified from the transgenic plants of T1 generation that came from 8 T0 transgenic plants with good resistance to aphid. The anti-aphid bioassay shows that resistance to the different grain aphid is not the same in transgenic wheat plants. To Rhopalosiphum padi, the rate of survival aphid 8 days after exposing transgenic plants to aphids is significantly lower than that of nontransgenic plants. To Macrosiphum avenae, growth speed of aphids is slowed down but not killed. At the same time, the death rate of young aphids is increased. Anyway, feeding of the two kinds of aphids has been controlled in a certain degree by gna gene when aphids can free to move in plants.