Search

The establishment of symbiosis between legume and rhizobium results in the formation of nodule. Phytocyanins (PCs) are a class of plant-specific blue copper proteins, playing critical roles in plant development including nodule formation. Although a few PC genes have been isolated from nodules, their functions are still unclear. Here, we performed a genome-wide identification of PC family in seven sequenced legume species (Medicago truncatula, Glycine max, Cicer arietinum, Cajanus cajan, Lotus japonicus, Vigna angularis and Phaseolus vulgaris) and found PCs experienced a remarkable expansion in M. truncatula and G. max. Further, we conducted an in-depth analysis of PC family in the model legume M. truncatula. Briefly, 82 MtPCs were divided into four subfamilies and clustered into seven clades, with a large proportion of tandem duplications and various cross-tissues expression patterns. Importantly, some PCs, such as MtPLC1, MtENODL27 and MtENODL28 were preferentially expressed in nodules. Further, RNA interference (RNAi) experiment revealed the knockdown of MtENDOL27 and MtENDOL28 impaired rhizobia infection, nodule numbers and nitrogenase activity. Moreover, in the MtENODL27-RNAi nodules, the infected cells were reduced and the symbiosomes did not reach the elongated stage, indicating MtENDOL27 is required for rhizobia infection and nodule development. In addition, co-expression analysis showed MtPLC1, MtENODL27 and MtENODL28 were grouped into two different functional modules and co-expressed with the known symbiotic nitrogen fixation-related genes, suggesting that they might participate in nodulation via different ways. In summary, this study provides a useful resource for future researches on the structure and function of PCs in nodulation.

Leaf necrosis induced by fungal pathogens is one of the most devastating diseases of yam. The objectives of this study were to examine the production of defensive phytohormones and perform a comparative transcriptome analysis between two yam cultivars with different resistance levels against Botrytis cinerea inoculation. After inoculation with B. cinerea, the endogenous ethylene level was found to have accumulated to a higher level in the MH1 resistant cultivar. Meanwhile, expression profiles identified differential defense regulation of ethylene pathway in MH1 (versus susceptible cv. MH3) in response to B. cinerea. A number of ethylene-synthesis and -responsive genes were expressed at higher levels in MH1 than in MH3 after inoculation. Furthermore, ethylene supplementation in MH3 plants by ethephon spraying indeed enhanced the resistance against B. cinerea and Colletotrichum alatae, and elevated the expression of DaEIN2 and DaERF96. Our work improves understanding of defense mechanism and highlights the function of ethylene potentially utilized for yam protection against diseases.

Tick-borne encephalitis virus (TBEV) is one of the flaviviruses that targets the CNS and causes encephalitis in humans. The mechanism of TBEV that causes CNS destruction remains unclear. It has been reported that RANTES-mediated migration of human blood monocytes and T lymphocytes is specifically induced in the brain of mice infected with TBEV, which causes ensuing neuroinflammation and may contribute to brain destruction. However, the viral components responsible for RANTES induction and the underlying mechanisms remain to be fully addressed. In this study, we demonstrate that the NS5, but not other viral proteins of TBEV, induces RANTES production in human glioblastoma cell lines and primary astrocytes. TBEV NS5 appears to activate the IFN regulatory factor 3 (IRF-3) signaling pathway in a manner dependent on RIG-I/MDA5, which leads to the nuclear translocation of IRF-3 to bind with RANTES promoter. Further studies reveal that the activity of RNA-dependent RNA polymerase (RdRP) but not the RNA cap methyltransferase is critical for TBEV NS5–induced RANTES expression, and this is likely due to RdRP-mediated synthesis of dsRNA. Additional data indicate that the residues at K359, D361, and D664 of TBEV NS5 are critical for RdRP activity and RANTES induction. Of note, NS5s from other flaviviruses, including Japanese encephalitis virus, West Nile virus, Zika virus, and dengue virus, can also induce RANTES expression, suggesting the significance of NS5-induced RANTES expression in flavivirus pathogenesis. Our findings provide a foundation for further understanding how flaviviruses cause neuroinflammation and a potential viral target for intervention.

Ecological stoichiometry has been widely studied in terrestrial ecosystems, but these studies have been limited in terms of symbiotic association between alfalfa and arbuscular mycorrhizal fungi (AMF), especially during regrowth. To evaluate the effect of AMF on the regrowth and C:N:P stoichiometry of alfalfa (Medicago sativa L.) under well-watered and drought conditions, alfalfa plants inoculated with AMF (Rhizophagus irregularis, M), nitrogen-fixing bacteria (Sinorhizobium, R), both nitrogen-fixing bacteria and AMF or no inoculations (CK) were evaluated in a pot experiment under controlled conditions. The biomass and organic carbon (C), nitrogen (N) and phosphorus (P) nutritional status of plant leaves and roots were measured under two water treatments during regrowth. Water deficit reduced the accumulation of dry matter and the concentrations of C and N in leaves and P in roots but increased the concentrations of P in leaves and C and N in roots of alfalfa during regrowth. Compared to CK plants, inoculation significantly improved the regrowth biomass and the concentrations of C, N and P in the leaves and roots and especially increased P levels when the plant were inoculated with AMF. However, this effect of microbes on alfalfa regrowth was dependent on the soil water status. Drought reduced the C:N and C:P in the leaves and the C:N in roots, while N:P and C:P increased in the roots. Inoculation of AMF decreased the C:P and N:P in the leaves and the C:N and C:P in the roots, whereas it increased the C:N under water stress. These results indicate that AMF play a significant role in regrowth and C:N:P ecological stoichiometry after defoliation by influencing C assimilation, N and P uptake and that the responses in the leaves and the roots are opposite.

Glutathione S-transferases are important enzymes in protecting cells by scavenging reactive oxygen species induced by various stresses. In this study, a novel GST gene, MsGST (KM044312), was cloned and characterized from alfalfa. The open reading frame of MsGST contains 660bp nucleotides, encoding 219 amino acid residues. Amino acid sequence alignment indicated that the deduced MsGST protein was highly homologous to other plant tau class GST sequences. According to amino acid phylogenetic analysis, the MsGST gene was clustered into the same branch with other legume plants. Real-time quantitative PCR (qRT-PCR) revealed that the expression levels of MsGST were up-regulated in both shoots and roots under ABA treatment and various stresses, including salt, drought, cold and heat stress. The effect of nodules on MsGST gene expression indicated that the induction of MsGST expression by abiotic stress is independent of rhizobium symbiosis. In conclusion, the MsGST gene may be involved in response to different abiotic stresses in alfalfa.