Citrus canker, caused by Xanthomonas axonopodis pv. citri (‘Xac’), is an important quarantine disease in citrus crops. Arbuscular mycorrhizal fungi (AMF) form symbiotic interactions with host plants and further affect their disease resistance, possibly by modulating the activity of salicylic acid (SA), a key phytohormone in disease resistance. Common mycorrhizal networks (CMNs) can interconnect plants, but it is not yet clear whether CMNs promote resistance to citrus canker and, if so, whether SA signaling is involved in this process. To test this possibility, we used a two‐chambered rootbox to establish CMNs between trifoliate orange (Poncirus trifoliata) seedlings in chambers inoculated (treated) or not (neighboring) with the AMF, Paraglomus occultum. A subset of the AMF‐inoculated seedlings were also inoculated with Xac (+AMF+Xac). At 2 d post‐inoculation (dpi), compared with the +AMF−Xac treatment, neighboring seedlings in +AMF+Xac treatment had lower expression levels of the SA biosynthetic genes, PtPAL, PtEPS1, and PtPBS3, but higher SA levels, which attributed to the upregulation of PtPAL and PtPBS3 in treated seedlings and the transfer of SA, via CMNs, to the neighboring seedlings. At 4 dpi, the pathogenesis‐related (PR) protein genes, PtPR1, PtPR4, and PtPR5, and the transcriptional regulatory factor gene, PtNPR1, were activated in neighboring seedlings of +AMF+Xac treatment. At 9 dpi, root phenylalanine ammonia‐lyase activity and total soluble phenol and lignin concentrations increased in neighboring seedlings of +AMF+Xac treatment, likely due to the linkage and signal transfer, via CMNs. These findings support the hypothesis that CMNs transfer the SA signal from infected to neighboring healthy seedlings, to activate defense responses and affording protection to neighboring plants against citrus canker infection.

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Citrus canker is a quarantined disease, severely harming citrus plants. Soil beneficial arbuscular mycorrhizal fungi (AMF) can provide a biological control pathway to resist pathogens. This work was to test changes of signal substances including hydrogen peroxide (H2O2), nitric oxide (NO), calmodulin (CaM), salicylic acid (SA) and jasmonic acid (JA) and the pathogen defense gene expression in roots of AMF (Paraglomus occultum) and non-AMF trifoliate orange (Poncirus trifoliata) seedlings after infected by a expressions citrus canker pathogen (Xanthomonas axonopodis pv. Citri, Xac). AMF inoculation significantly improved plant height, stem diameter and leaf number. Xac infection dramatically decreased root H2O2, NO, and SA levels, but increased root CaM and JA concentrations in non-AMF seedlings. There were higher H2O2 and CaM levels and lower JA levels in Xac-infected seedlings than in non-Xac-infected seedlings under mycorrhization. Under non-Xac infection, mycorrhizal treatment reduced root H2O2, NO, and SA but increased CaM and JA levels. However, under Xac infection, mycorrhizal inoculation distinctly accelerated root H2O2, NO, CaM, and SA accumulation, accompanied with up-regulated expression levels of root PtEPS1 (enhance pseudomonas susceptibility 1) and PtPR4 (pathogenesis related gene 4), indicating that Xac stimulated mycorrhizal roles in enhancing resistance of citrus canker. Such results imply that citrus plants with pre-inoculated AMF had stronger resistance to Xac infection through increasing signal substrate accumulation and pathogen defense gene expressions.   ********* In press - Online First. Article has been peer reviewed, accepted for publication and published online without pagination. It will receive pagination when the issue will be ready for publishing as a complete number (Volume 47, Issue 4, 2019). The article is searchable and citable by Digital Object Identifier (DOI). DOI link will become active after the article will be included in the complete issue. *********

Common mycorrhizal networks (CMNs) connecting two or more neighbouring plants are confirmed to transfer signals, whereas little information about CMNs effects on the signal substances production is known. In this study, a two-chambered rootbox separated by 37 µm nylon mesh was used to establish donor and receptor chambers. Two chambers both were planted with trifoliate orange (Poncirus trifoliata) and then only donor chamber inoculated with Diversispora versiformis, Paraglomus occultum and Rhizoglomus intraradices. The roots of the donor and receptor plants both were mycorrhizated suggesting that CMNs were established between donor and receptor seedlings. Moreover, the AMF association dramatically increased plant height, stem diameter, leaf numbers, and shoot and root biomass in both the donor and receptor seedlings. The AMF inoculation in the donor plants and the subsequent mycorrhizal colonization by CMNs in the receptor plants significantly increased root calmodulin (CaM) and salicylic acid (SA) concentrations, while considerably decreased root nitric oxide (NO) and jasmonic acid (JA) concentrations. This was accompanied by down-regulated expression of three JA synthetic genes (PtLOX, PtAOS and PtAOC), regardless of donor and receptor seedlings. These results thus suggest that CMNs between trifoliate orange seedlings manifestly promote plant growth and affect the production of signal substances.