Dark septate endophytes (DSEs) are one of the most studied groups of root fungal endophytes in recent years. However, the effects of DSE on host plant are still under debate, and the molecular mechanisms are poorly understood. In this study, we identified a DSE fungus of the genus Anteaglonium, named T010, from the wild blueberry. When inoculated into Vaccinium corymbosum L. plants, T010 could enhance root growth and promote shoot branching, leading to increased plant growth. By comparative transcriptome analysis, we obtained 1948 regulated differentially expressed genes (DEGs) from the V. corymbosum plants treated by T010. Further functional enrichment analysis identified a series of DEGs enriched in transcriptional regulation, material transport, phytohormone biosynthesis and flavonoid biosynthesis. Moreover, the comparative analysis of liquid chromatography–mass spectrometry verified that T010 treatment induced the changes in the contents of various phytohormones and flavonoids. This is the first report on the isolation of DSE fungi of the genus Anteaglonium from blueberry roots. Moreover, our results suggested that T010 colonization could result in a series of changes in cell metabolism, biosynthesis and signal pathways, thereby promoting plant growth. Particularly, the changes of phytohormone and flavonoid metabolism induced by T010 colonization might contribute to the promotion of blueberry growth. Our results will provide new insights into understanding of the interaction of DSE fungi and host plants, as well as the development and utilization of DSE preparations.

Ser/Thr phosphorylation by serine/threonine protein kinases (STPKs) plays significant roles in molecular regulation, which allows Mycobacteria to adapt their cell wall structure in response to the environment changes. Identifying direct targets of STPKs and determining their activities are therefore critical to revealing their function in Mycobacteria, for example, in cell wall formation and virulence. Herein, we reported that RmlA, a crucial L-rhamnose biosynthesis enzyme, is a substrate of STPK PknB in Mycobacterium tuberculosis (M. tuberculosis). Mass spectrometry analysis revealed that RmlA is phosphorylated at Thr-12, Thr-54, Thr-197, and Thr-12 is located close to the catalytic triad of RmlA. Biochemical and phenotypic analysis of two RmlA mutants, T12A/T12D, showed that their activities were reduced, and cell wall formation was negatively affected. Moreover, virulence of RmlA T12D mutant was attenuated in a macrophage model. Overall, these results provide the first evidence for the role of PknB-dependent RmlA phosphorylation in regulating cell wall formation in Mycobacteria, with significant implications for pathogenicity.

Drought is a significant global issue affecting agricultural production, and the utilization of beneficial rhizosphere microorganisms is one of the effective ways to increase the productivity of crops and forest under drought. In this study, we characterized a novel growth-promoting dark septate endophytes (DSE) fungus R16 (Dothideomycetes sp.) derived from blueberry roots. Hyphae or microsclerotia were visible within the epidermal or cortical cells of R16-colonized blueberry roots, which was consistent with the typical characteristics of DSE fungi. Inoculation with R16 promoted the growth of blueberry seedlings, and the advantage over the control group was more significant under PEG-induced drought. Comparison of physiological indicators related to drought resistance between the inoculated and control groups was performed on the potted blueberry plants, including the chlorophyll content, net photosynthetic rate, root activities, malondialdehyde and H2O2 content, which indicated that R16 colonization mitigated drought injury in blueberry plants. We further analyzed the effects of R16 on phytohormones and non-structural carbohydrates (NSCs) to explore the mechanism of increased drought tolerance by R16 in blueberry seedlings. The results showed that except for the gibberellin content, indole-3-acetic acid, zeatin and abscisic acid varied significantly between the inoculated and control groups. Sucrose phosphate synthase and sorbitol-6-phosphate dehydrogenase activities in mature leaves, the key enzymes responsible for sucrose and sorbitol synthesis, respectively, as well as sorbitol dehydrogenase, sucrose synthase, cell wall invertase, hexokinase and fructokinase in roots, the key enzymes involved in the NSCs metabolism, showed significant differences between the inoculated and control groups before and after drought treatment. These results suggested that the positive effects of R16 colonization on the drought tolerance of blueberry seedlings are partially attributable to the regulation of phytohormone and sugar metabolism. This study provided valuable information for the research on the interaction between DSE fungi and host plants as well as the application of DSE preparations in agriculture.

Lectin receptor-like kinases (LecRLKs), a large family of plant receptor-like kinases (RLKs), play an important role in plant response to abiotic stresses. However, little information is available about the roles of LecRLKs in the salt stress response of sweet cherry (Prunus avium). Here, an L-type LecRLK gene (PaLectinL7) was characterized from sweet cherry. Subcellular localization analysis revealed that PaLectinL7 was a plasma membrane protein. The expression of PaLectinL7 was up-regulated by salt, drought and exogenously gibberellin (GA3) treatments. Overexpression of PaLectinL7 in the roots of Gisela 6 enhanced its tolerance to salt stress. Additionally, transcriptome analysis showed that lignin metabolic related genes were regulated by PaLectinL7 overexpression. Meanwhile, the lignin contents and associated enzymes (CAD and COMT) rose concurrently with PaLectinL7 overexpression under salt stress. We also found that PaCAD1, a key enzyme involved in lignin metabolism, interacted with PaLectinL7 and could be phosphorylated by PaLectinL7 in vitro, suggesting that PaLectinL7 may regulate the enzyme activity of PaCAD1. Therefore, these results indicated that PaLectinL7, as a membrane-bound regulator, promoted lignin deposition by regulating the activities of enzymes related to lignin metabolism, thus enhancing salt tolerance.