Stroke remains one of the leading causes of permanent disability and death worldwide. Apoptosis and autophagy are two key elements involved in ischemic brain damage. Ethanol is a commonly used and abused chemical substance that affects the prognosis of ischemic stroke. We determined the influence of chronic ethanol consumption on apoptosis and autophagy following transient focal cerebral ischemia. Male C57BL/6 J mice were randomly divided into three groups and gavage fed with 0.7 and 2.8 g/kg/day ethanol or volume-matched water daily for 8 weeks. DNA fragmentation, TUNEL-positive neurons, cleaved caspase-3-positive neurons, translocation of mitochondrial cytochrome C and apoptosis inducing factor (AIF), LC3B-positive neurons, and expression of LC3B, Beclin-1 and Bcl-2 in peri-infarct cortex were evaluated at 24 hours of reperfusion after a 90-minute unilateral middle cerebral artery occlusion (MCAO). Cerebral ischemia/reperfusion (I/R) injury was significantly improved in the 0.7 g/kg/d ethanol group but worsened in the 2.8 g/kg/d ethanol group. DNA fragmentation was significantly increased at 24 hours of reperfusion in all groups. However, the magnitude of the increase was significantly less in the 0.7 g/kg/d ethanol group. In addition, both cleaved caspase-3-positive neurons and TUNEL-positive neurons were significantly less in 0.7 g/kg/d ethanol group. Furthermore, translocation of mitochondrial cytochrome C and AIF was significantly alleviated in the 0.7 g/kg/d ethanol group. On the other hand, baseline expression of LC3B was significantly reduced in the 2.8 g/kg/d ethanol group. Post-ischemic expression of LC3B and LC3B-positive neurons were significantly attenuated in both 0.7 and 2.8 g/kg/d ethanol groups. Moreover, although post-ischemic expression of Beclin-1 was not altered in the ethanol groups, post-ischemic expression of Bcl-2 was significantly greater in both 0.7 and 2.8 g/kg/d ethanol groups. Our findings suggest that light ethanol consumption may protect against cerebral I/R injury by suppressing post-ischemic apoptosis, whereas heavy ethanol consumption may exacerbate cerebral I/R injury by suppressing autophagy.

The Mildew Resistance Locus (Mlo) gene family is reported in various species as regulators of powdery mildew (PM) resistance. However, the Mlo genes in cucurbit crops remain limited. In this study, a genome-wide investigation of Mlo genes was conducted in eight Cucurbitaceae species and in rice, maize, arabidopsis, and barley, and a total of 202 Mlo genes were identified. The phylogenetic analysis showed that 202 Mlo genes can be classified into six clades, and the Mlo genes from clades I and III are likely pivotal in regulating PM resistance in dicotyledonous and monocotyledonous plants, respectively. The Ka/Ks ratios for these homologous Mlo gene pairs ranged from 0 to 0.6, revealing that they underwent substantial purifying selection during evolution. Among 12 crops, there were the most Mlo genes (22 Cm-mlo) in melon. An expression analysis revealed that six Cm-mlo genes showed expression responses to PM infection in which Cm-mlo38 and Cm-mlo44 were phylogenetically close to Mlo genes that regulated PM resistance. Using the VIGS system for silencing, Cm-mlo38 and Cm-mlo44 enhanced resistance to PM in susceptible material. A protein interaction analysis indicated that Cm-mlo38 might regulate PM resistance through interactions with PR5 and CML proteins. These results provide a comprehensive understanding of the Mlo family in Cucurbitaceae and pave the way for future functional analysis and genetic improvement for improving PM resistance in Cucurbitaceae.

Calmodulin-binding transcription activator (CAMTA) is an important calmodulin-binding protein with a conserved structure in eukaryotes which is widely involved in plant stress response, growth and development, hormone signal transduction, and other biological processes. Although CAMTA genes have been identified and characterized in many plant species, a systematic and comprehensive analysis of CAMTA genes in the Solanaceae genome is performed for the first time in this study. A total of 28 CAMTA genes were identified using bioinformatics tools, and the biochemical/physicochemical properties of these proteins were investigated. CAMTA genes were categorized into three major groups according to phylogenetic analysis. Tissue-expression profiles indicated divergent spatiotemporal expression patterns of SmCAMTAs. Furthermore, transcriptome analysis of SmCAMTA genes showed that exposure to cold induced differential expression of many eggplant CAMTA genes. Yeast two-hybrid and bimolecular fluorescent complementary assays suggested an interaction between SmCAMTA2 and SmERF1, promoting the transcription of the cold key factor SmCBF2, which may be an important mechanism for plant cold resistance. In summary, our results provide essential information for further functional research on Solanaceae family genes, and possibly other plant families, in the determination of the development of plants.

Studying the interrelationship between agroecosystems and the microbiome is essential in achieving sustainable agricultural development. In particular, the relationship between the rhizosphere microbiome and crops is critical to the health and stability of agroecosystems. This study evaluated the diversity and keystone species of the cucumber rhizosphere microbiome and investigated their contributing role in cucumber growth and microbial community regulation. A total of 108 soil samples were collected from the rhizosphere and bulk soil of cucumber crops in different locations and under different cultivation modes in a high-yield area. These samples were analyzed to study the interrelationships between different cultivation modes and crop rhizosphere microorganisms. The results revealed significant regional differences in microbial communities due to microhabitat (rhizosphere and bulk soil) and geographic location. Common bacteria such as Pseudomonas, Sphingomonas, Lysobacter and Rhodanobacter were the most abundant and showed strong correlations with several other bacteria through co-occurrence networking. In addition, Flavobacterium sp. and Bacillus sp. were identified as keystone species for the rhizobia community structure. Synthetic microbial communities and pot experiments confirmed that these two keystone species could enrich the soil microbime and promote cucumber growth. Flavobacterium sp. may promote cucumber growth by regulating the rhizosphere microbial community while Bacillus sp. may influence cucumber growth by stabilizing the rhizosphere microbial biomass, further enhancing nutrient cycling efficiency and improving system stability, which in turn affected the agroecosystem. Our study demonstrates that specific keystone species are important in shaping the cucumber rhizosphere microbiome and plant growth. These results provide insights into agroecosystem-microbe interrelationships and provide a potential basis for optimizing agricultural management strategies.