The septins FaCdc3 and FaCdc12 are required for cytokinesis and affect asexual and sexual development,lipid metabolism and virulence in Fusarium asiaticum

Septins are a highly conserved family of GTP‐binding proteins that contribute to many cellular and metabolic functions, including cell polarity, cytokinesis, cell morphogenesis and pathogenesis. In this study, we characterized the septins FaCdc3 and FaCdc12 in the filamentous fungus Fusarium asiaticum. The functions of FaCdc3 and FaCdc12 were evaluated by constructing deletion mutants of FaCdc3 and FaCdc12, designated ΔFaCdc3‐5 and ΔFaCdc12‐71, respectively. The deletion mutants exhibited a reduced rate of mycelial growth, increased aerial hyphae formation, irregularly shaped hyphae, reduced conidiation and a lack of sexual reproduction in wheat kernels. Histochemical analysis revealed that the conidia and hyphae of ΔFaCdc3‐5 and ΔFaCdc12‐71 formed large lipid droplets (LDs). ΔFaCdc3‐5 and ΔFaCdc12‐71 also exhibited increased resistance to agents that induce osmotic stress and damage the cell membrane and cell wall. In addition, the hyphae and conidia of the two mutants formed fewer septa than those of the wild‐type and exhibited aberrant nuclear distribution. Pathogenicity assays showed that ΔFaCdc3‐5 and ΔFaCdc12‐71 exhibited reduced virulence on wheat spikelets, which was indirectly correlated with a reduced level of deoxynivalenol accumulation. All of these defects were restored by genetic complementation of the two mutants with the parental FaCdc3 and FaCdc12. These results indicate that FaCdc3 and FaCdc12 play a critical role in various cellular processes in F. asiaticum.     

秦岭细鳞鲑栖息地环境特征研究

为研究秦岭细鳞鲑（Brachymystax lenok tsinlingensis）的栖息地环境选择偏好，对陕西陇县秦岭细鳞鲑国家级自然保护区和甘肃秦州珍稀水生野生动物国家级自然保护区内25个样点进行了鱼类采样和生境测量。共采集到130尾秦岭细鳞鲑样本，样点的平均分布密度为（0.10±0.07）尾/m，两个保护区内秦岭细鳞鲑的密度存在显著性差异（P < 0.05）。与秦岭细鳞鲑密度分布呈正相关的因子依次为坡度、跌水区密度、海拔、粗糙度、遮蔽度、底栖动物生物量和流速，与密度呈负相关的环境因子依次为河宽、水深和溶氧。冗余性分析（RDA）筛选了坡度、粗糙度、遮蔽度、海拔和跌水区5个关键环境因子。基于关键环境因子的实测值建立了栖息地适合度曲线，秦岭细鳞鲑分布的最适坡度范围为5%-10%，海拔分布范围为1500-2000 m，粗糙度范围0.3-0.4；跌水区密度范围12-18个/100 m，遮蔽度在0.5以上。     

The EF‐1α promoter maintains high‐level transgene expression from episomal vectors in transfected CHO‐K1 cells

In our previous study, we demonstrated that episomal vectors based on the characteristic sequence of matrix attachment regions (MARs) and containing the cytomegalovirus (CMV) promoter allow transgenes to be maintained episomally in Chinese hamster ovary (CHO) cells. However, the transgene expression was unstable and the number of copies was low. In this study, we focused on enhancers, various promoters and promoter variants that could improve the transgene expression stability, expression magnitude (level) and the copy number of a MAR‐based episomal vector in CHO‐K1 cells. In comparison with the CMV promoter, the eukaryotic translation elongation factor 1 α (EF‐1α, gene symbol EEF1A1) promoter increased the transfection efficiency, the transgene expression, the proportion of expression‐positive clones and the copy number of the episomal vector in long‐term culture. By contrast, no significant positive effects were observed with an enhancer, CMV promoter variants or CAG promoter in the episomal vector in long‐term culture. Moreover, the high‐expression clones harbouring the EF‐1α promoter tended to be more stable in long‐term culture, even in the absence of selection pressure. According to these findings, we concluded that the EF‐1α promoter is a potent regulatory sequence for episomal vectors because it maintains high transgene expression, transgene stability and copy number. These results provide valuable information on improvement of transgene stability and the copy number of episomal vectors.