cBacillus cyclic lipopeptides (LPs) have been well studied for their phytopathogen-antagonistic activities. Recently, research has shown that these LPs also contribute to the phenotypic features of Bacillus strains, such as hemolytic activity, swarming motility, biofilm formation, and colony morphology. Bacillus subtilis 916 not only coproduces the three families of well-known LPs, i.e., surfactins, bacillomycin Ls (iturin family), and fengycins, but also produces a new family of LP called locillomycins. The genome of B. subtilis 916 contains four nonribosomal peptide synthase (NRPS) gene clusters, srf, bmy, fen, and loc, which are responsible for the biosynthesis of surfactins, bacillomycin Ls, fengycins, and locillomycins, respectively. By studying B. subtilis 916 mutants lacking production of one, two, or three LPs, we attempted to unveil the connections between LPs and phenotypic features. We demonstrated that bacillomycin Ls and fengycins contribute mainly to antifungal activity. Although surfactins have weak antifungal activity in vitro, the strain mutated in srfAA had significantly decreased antifungal activity. This may be due to the impaired productions of fengycins and bacillomycin Ls. We also found that the disruption of any LP gene cluster other than fen resulted in a change in colony morphology. While surfactins and bacillomycin Ls play very important roles in hemolytic activity, swarming motility, and biofilm formation, the fengycins and locillomycins had little influence on these phenotypic features. In conclusion, B. subtilis 916 coproduces four families of LPs which contribute to the phenotypic features of B. subtilis 916 in an intricate way.

The antagonistic activity of lipopeptides in Bacillus subtilis 916 has been well documented, yet relatively little is known about their mechanism in biofilm formation and environmental colonization. This study sought to examine the interaction of B. subtilis 916 on Rhizoctonia solani-infected rice sheath to elucidate the mechanism of colonization on plant leaves. Results showed that the mutants Δbac, Δsrf, and Δsrf + bac of B. subtilis 916, deficient in bacillomycin L and surfactin production, respectively, not only altered colony morphology but also changed swarming motility, reduced antagonistic activity, and decreased biofilm formation. In particular, biofilm formation in mutant Δbac, not Δsrf or Δsrf + bac, were restored with addition of surfactin and bacillomycin L at 10 and 50 μg/mL, respectively. Moreover, surfactin and bacillomycin L were able to restore or enhance swarming motility in the corresponding mutants at 10 μg/mL, respectively. With the aid of green fluorescent protein tagging, it was demonstrated that B. subtilis 916 formed a robust biofilm on the rice sheath blight lesion and colonized well on R. solani-infected rice sheath, while its corresponding mutants performed poorly. These observations also correlated with the rice cultivar pot experiments, in which B. subtilis 916 exhibited greater biocontrol than its mutants. Our results suggest that surfactin and bacillomycin L contribute differently but synergistically to the biocontrol of rice sheath blight in B. subtilis 916 through its antifungal activity, biofilm formation, and colonization.

Domain of unknown function 1644 (DUF1644) is a highly conserved amino acid sequence motif present only in plants. Analysis of expression data of the family of DUF1644-containing genes indicated that they may regulate responses to abiotic stress in rice. Here we present our discovery of the role of OsSIDP366, a member of the DUF1644 gene family, in response to drought and salinity stresses in rice. Transgenic rice plants overexpressing OsSIDP366 showed enhanced drought and salinity tolerance and reduced water loss as compared to that in the control, whereas plants with downregulated OsSIDP366 expression levels using RNA interference (RNAi) were more sensitive to salinity and drought treatments. The sensitivity to abscisic acid (ABA) treatment was not changed in OsSIDP366-overexpressing plants, and OsSIDP366 expression was not affected in ABA-deficient mutants. Subcellular localization analysis revealed that OsSIDP366 is presented in the cytoplasmic foci that colocalized with protein markers for both processing bodies (PBs) and stress granules (SGs) in rice protoplasts. Digital gene expression (DGE) profile analysis indicated that stress-related genes such as SNAC1, OsHAK5 and PRs were upregulated in OsSIDP366-overexpressing plants. These results suggest that OsSIDP366 may function as a regulator of the PBs/SGs and positively regulate salt and drought resistance in rice.

Remote ischemic conditioning has been convincingly shown to render the myocardium resistant to a subsequent more severe sustained episode of ischemia. Compared with other organs, little is known regarding the effect of transient liver ischemic conditioning. We proposed the existence of cardioprotection induced by remote liver conditioning. Male Sprague-Dawley rats were divided into sham-operated control (no further hepatic intervention) and remote liver ischemic conditioning groups. For liver ischemic conditioning, three cycles of 5 min of liver ischemia-reperfusion stimuli were conducted before-(liver preconditioning), post-myocardial ischemia (liver postconditioning), or in combination of both (liver preconditioning + liver postconditioning). Rats were exposed to 45 min of left anterior descending coronary artery occlusion, followed by 3 h of reperfusion thereafter. ECG and hemodynamics were measured throughout the experiment. The coronary artery was reoccluded at the end of reperfusion for infarct size determination. Blood samples were taken for serum lactate dehydrogenase and creatine kinase-MB test. Heart tissues were taken for apoptosis measurements and Western blotting. Our data demonstrate that liver ischemic preconditioning, postconditioning, or a combination of both, offered strong cardioprotection, as evidenced by reduction in infarct size and cardiac tissue damage, recovery of cardiac function, and inhibition of apoptosis after ischemia-reperfusion. Moreover, liver ischemic conditioning increased cardiac (not hepatic) glycogen synthase kinase-3β (GSK-3β) phosphorylation. Accordingly, inhibition of GSK-3β mimicked the cardioprotective action of liver conditioning. These results demonstrate that remote liver ischemic conditioning protected the heart against ischemia and reperfusion injury via GSK-3β-dependent cell-survival signaling pathway. Remote ischemic conditioning protects hearts against ischemia and reperfusion (I/R) injury. However, it is unclear whether ischemic conditioning of visceral organs such as the liver, the largest metabolic organ in the body, can produce cardioprotection. This is the first study to show the cardioprotective effect of remote liver ischemic conditioning in a rat model of myocardial I/R injury. We also, for the first time, demonstrated these protective properties are associated with glycogen synthase kinase-3β-dependent cell-survival signaling pathway.

Domain of Unknown Function 966 (DUF966) gene family was found in the protein family database, which consisted of seven genes in rice. The proteins encoded by these genes contained one or two highly conserved DUF966 domains. The available data of public microarray databases implied that these genes might play crucial roles in plant response to abiotic stresses. In this study, a member of the DUF966 gene family, DUF966-stress repressive gene 2 in Oryza sativa (OsDSR2, Loc_Os01g62200), was cloned and its role in rice responding to salt and simulated drought stresses was functionally characterized. OsDSR2 was expressed mainly in nodes of stems and leaf blades from rice. Expression profile analysis of adversity showed that OsDSR2 had different transcriptional responses to salt, drought, cold, heat and oxidative (H2O2) stresses, as well as abscisic acid (ABA), methyl jasmonate, salicylic acid, gibberellin acid and auxin treatments. Transient expression demonstrated that OsDSR2 was localized in the membrane and nucleus. Overexpression of OsDSR2 could increase salt and simulated drought (polyethyleneglycol)-stress sensitivities in rice by downregulating the expression of ABA- and stress-responsive genes including OsNCED4, SNAC1, OsbZIP23, P5CS, Oslea3 and rab16C. Furthermore, OsDSR2-overexpressing plants showed reduced ABA sensitivity during the post-germination stage. These results suggested that OsDSR2 negatively regulated rice response to salt and simulated drought stresses as well as ABA signaling, which provided some useful data for understanding the functional roles of DUF966 family genes in abiotic stress responses in plants.

MicroRNAs (miRNAs) play a pivotal role in post-transcriptional regulation of gene expression in plants. In this study, we investigate miRNAs in an agronomically important common tobacco in China, named Honghua Dajinyuan (a drought-tolerant cultivar). Here, we report a comprehensive analysis of miRNA expression profiles in mock-treat grown (CK) and 20 % polyethylene glycol-grown (PEG-grown) tobacco roots using a high-throughput sequencing approach. A total of 656 unique miRNAs representing 53 miRNA families were identified in the two libraries, of which 286 unique miRNAs representing 162 microRNAs were differentially expressed. In addition, nine differentially expressed microRNAs selected from different expressed miRNA family with high abundance were subjected to further analysis and validated by quantitative real-time PCR (Q-PCR). In addition, the expression pattern of these identified candidate conserved miRNA and target genes of three identified miRNA (nta-miR172b, nta-miR156i, and nta-miR160a) were also validated by Q-PCR. Gene ontology (GO) enrichment analysis suggests that the putative target genes of these differentially expressed miRNAs are involved in metabolic process and response to stimulus. In particular, 25 target genes are involved in regulating plant hormone signal transduction and metabolism, indicating that these association microRNAs may play important regulatory roles in responding to PEG resistance. Moreover, this study adds a significant number of novel miRNAs to the tobacco miRNome.

Oligodendrocytes form myelin sheaths around axons in the central nervous system. Although cholesterol is one of the major lipid components in myelin sheath, the source of cholesterol for myelination remains to be defined. In this study, we report that low-density lipoprotein receptor (LDLR) and very low-density lipoprotein receptor (VLDLR) are selectively expressed in mature myelinating oligodendrocytes in the postnatal CNS. Both receptors are specifically expressed in differentiated oligodendrocytes in P7 spinal cord, but progressively down-regulated after P15. In adult animals, only LDLR expression can be detected in a small number of oligodendrocytes throughout the entire spinal cord. In the brain region, LDLR is expressed by the white matter oligodendrocytes of both cerebellum and cerebral cortex, whereas VLDLR has a weak expression in cerebellar oligodendrocytes. Together, our expression studies suggest that cholesterol uptake by LDLR and VLDLR may play an important role in the formation of myelin sheath.

Many studies have reported that increasing atmospheric nitrogen (N) deposition broadens N:phosphorus (P) in both soils and plant leaves and potentially intensifies P limitation for plants. However, few studies have tested whether P addition alleviates N-induced P limitation for plant belowground growth. It is also less known how changed N:P in soils and leaves affect plant belowground stoichiometry, which is significant for maintaining key belowground ecological processes. We conducted a multi-level N:P supply experiment (varied P levels combined with constant N amount) for Glycyrrhiza uralensis (a N fixing species) and Pennisetum centrasiaticum (a grass) from a desert steppe in Northwest China during 2011–2013. Results showed that increasing P addition increased the belowground biomass and P concentrations of both species, resulting in the decreases in belowground carbon (C):P and N:P. These results indicate that P inputs alleviated N-induced P limitation and hence stimulated belowground growth. Belowground C:N:P stoichiometry of both species, especially P. centrasiaticum, tightly linked to soil and green leaf C:N:P stoichiometry. Thus, the decoupling of C:N:P ratios in both soils and leaves under a changing climate could directly alter plant belowground stoichiometry, which will in turn have important feedbacks to primary productivity and C sequestration.