The proteins of Ras family are a large group of monomeric GTPases and act as molecular switches transducing extracellular signals into the cell in higher eukaryotes. However, little is known about roles of Ras family in the foliar pathogens. In this research, we cloned the gene named StRas2 encoding Ras in Setosphaeria turcica and investigated its function by RNA interference technology. We found that the growth rate of RNAi transformants named as R1, R2, R3, R4, R5 and R6, in which the StRas2 silencing efficiency fell in turn. With the highest silencing efficiency, the transformant R1 showed anomalistic hyphae morphology, indicating its growth was significantly affected. The transformants with a middle-silencing efficiency, such as R3, R4, displayed a delay when forming appressoria and invasive hyphae. R1 could not form conidia and appressoria. However, the conidial formation in R5 and R6 was significantly reduced, and these two transformants could form appressoria and penetrate the artificial cellophane, only that its invasive hyphae were fascicular and rarely branched. The HT-toxin biological activity of all transformants showed no difference. All results suggested that StRas2 is involved in the morphogenesis, conidiation, and appressorium development and is not related to the biosynthesis of HT-toxin.

No abstract

ObjectiveIn order to improve fermentation quality of natural grasses, their silage preparation and fermentation quality in meadow steppe (MS) and typical steppe (TS) were studied.MethodsThe small-scale silages and round bale silages of mixed natural grasses in both steppes were prepared using the commercial lactic acid bacteria (LAB) inoculants Chikuso-1 (CH, Lactobacillus plantarum) and cellulase enzyme (AC, Acremonium cellulase) as additives.ResultsMS and TS contained 33 and 9 species of natural grasses, respectively. Stipa baicalensis in MS and Stipa grandi in TS were the dominant grasses with the highest dry matter (DM) yield. The crude protein (CP), neutral detergent fiber and water-soluble carbohydrate of the mixed natural grasses in both steppes were 8.02% to 9.03%, 66.75% to 69.47%, and 2.02% to 2.20% on a DM basis, respectively. All silages treated with LAB and cellulase were well preserved with lower pH, butyric acid and ammonia-N content, and higher lactic acid and CP content than those of control in four kinds of silages. Compared with CH- or AC-treated silages, the CH+ AC-treated silages had higher lactic acid content.ConclusionThe results confirmed that combination with LAB and cellulase may result in beneficial effects by improving the natural grass silage fermentation in both grasslands.

In this study, we analyzed the fermentation quality, microbial community, and metabolome characteristics of ryegrass silage from different harvests (first harvest-AK, second harvest-BK, and third harvest-CK) and analyzed the correlation between fermentative bacteria and metabolites. The bacterial community and metabolomic characteristics were analyzed by single-molecule real-time (SMRT) sequencing and ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS/MS), respectively. After 60 days of ensiling, the pH of BK was significantly lower than those of AK and CK, and its lactic acid content was significantly higher than those of AK and CK. Lactiplantibacillus and Enterococcus genera dominate the microbiota of silage obtained from ryegrass harvested at three different harvests. In addition, the BK group had the highest abundance of Lactiplantibacillus plantarum (58.66%), and the CK group had the highest abundance of Enterococcus faecalis (42.88%). The most annotated metabolites among the differential metabolites of different harvests were peptides, and eight amino acids were dominant in the composition of the identified peptides. In the ryegrass silage, arginine, alanine, aspartate, and glutamate biosynthesis had the highest enrichment ratio in the metabolic pathway of KEGG pathway enrichment analysis. Valyl-isoleucine and glutamylvaline were positively correlated with Lactiplantibacillus plantarum. D-Pipecolic acid and L-glutamic acid were positively correlated with Levilactobacillus brevis. L-phenylalanyl-L-proline, 3,4,5-trihydroxy-6-(2-methoxybenzoyloxy) oxane-2-carboxylic acid, and shikimic acid were negatively correlated with Levilactobacillus brevis. In conclusion, this study explains the effects of different harvest frequencies on the fermentation quality, microbial community, and metabolites of ryegrass, and improves our understanding of the ensiling mechanisms associated with different ryegrass harvesting frequencies.

Silage exposed to air is prone to deterioration and production of unpleasant volatile chemicals that can seriously affect livestock intake and health. The aim of this study was to investigate the effects of Lactobacillus plantarum (LP), Lactobacillus buchneri (LB), and a combination of LP and LB (PB) on the quality, microbial community and volatile chemicals of Leymus chinensis silage at 0, 4, and 8 days after aerobic exposure. During aerobic exposure, LP had higher WSC and LA contents but had the least aerobic stability, with more harmful microorganisms such as Penicillium and Monascus and produced more volatile chemicals such as Isospathulenol and 2-Furancarbinol. LB slowed down the rise in pH, produced more acetic acid and effectively improved aerobic stability, while the effect of these two additives combined was intermediate between that of each additive alone. Correlation analysis showed that Actinomyces, Sphingomonas, Penicillium, and Monascus were associated with aerobic deterioration, and Weissella, Pediococcus, Botryosphaeria, and Monascus were associated with volatile chemicals. In conclusion, LB preserved the quality of L. chinensis silage during aerobic exposure, while LP accelerated aerobic deterioration.