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.

• MeSH
• kyselina salicylová metabolismus   MeSH
• Poncirus metabolismus   mikrobiologie   MeSH
• Xanthomonas axonopodis patogenita   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kyselina salicylová MeSH

Flavonoids are secondary metabolites widely found in plants with antioxidants, of which chalcone synthase (CHS) is a key enzyme required in flavonoid synthesis pathways. The objective of this study was to clone a CHS gene from trifoliate orange (Poncirus trifoliata) and analyze its biological information and partial functions. A PtCHS gene (NCBI accession: MZ350874) was cloned from the genome-wide of trifoliate orange, which has 1156 bp in length, encoding 391 amino acids, with a predicted protein relative molecular mass of 42640.19, a theoretical isoelectric point of 6.28, and a lipid coefficient of 89.82. The protein is stable, hydrophilic, and high sequence conservation (92.49% sequence homology with CHS gene of other species). PtCHS was highly expressed in stems, leaves and flowers, but very low expression in roots and seeds. Soil water deficit could up-regulate expressions of PtCHS in leaves. An arbuscular mycorrhizal fungus, Funneliformis mosseae, significantly increased plant biomass production, CHS activity, expressions of PtCHS, and total flavonoid content in leaves and roots, independent of soil water status. Total flavonoids were significantly positively correlated with PtCHS expression in leaves only and also positively with root mycorrhizal colonization. Such results provide insight into the important functions of PtCHS in trifoliate orange.

• Klíčová slova
• arbuscular mycorrhiza, chalcone synthase, drought, flavonoid, trifoliate orange,
• Publikační typ
• časopisecké články MeSH

Arbuscular mycorrhizas (AMs) have the ability to enhance drought tolerance of citrus, but the underlying mechanisms have not been clearly elucidated. Considering the strong association of cell membrane fatty acid (FA) unsaturation with plant drought tolerance, the present study hypothesized that AM fungi (AMF) modulated the composition and unsaturation of FAs to enhance drought tolerance of host plants. Drought-sensitive citrus rootstocks, trifoliate orange (Poncirus trifoliata) seedlings, were inoculated with AMF (Funneliformis mosseae) for 3 months and were subsequently exposed to drought stress (DS) for 8 weeks. Mycorrhizal seedlings exhibited better plant growth performance, higher leaf water potential and lower root abscisic acid concentrations under both well-watered (WW) and DS conditions. Arbuscular mycorrhiza fungus inoculation considerably increased root methyl oleate (C18:1), methyl linoleate (C18:2) and methyl linolenate (C18:3N3) concentrations under both WW and DS conditions, and root methyl palmitoleate (C16:1) concentrations under WW, while it decreased root methyl stearate (C18:0) levels under both WW and DS. These changes in the composition of FAs of mycorrhized roots resulted in higher unsaturation index of root FAs, which later aided in reducing the oxidative damage on account of lower concentration of malondialdehyde and superoxide radicals. The changes of these FAs were a result of AMF-up-regulating root FA desaturase 2 (PtFAD2), FA desaturase 6 (PtFAD6) and Δ9 FA desaturase (PtΔ9) genes under WW and PtFAD2, PtFAD6 and Δ15 FA desaturase (PtΔ15) genes under DS conditions. Our results confirmed that mycorrhization brought significant changes in root FA compositions, in addition to regulation of gene expression responsible for increasing the unsaturation level of FAs, a predisposing physiological event for better drought tolerance of citrus.

• Klíčová slova
• Poncirus trifoliata, drought, mycorrhizal symbiosis, unsaturation index of fatty acids,
• MeSH
• Citrus * MeSH
• Glomeromycota * MeSH
• kořeny rostlin MeSH
• mykorhiza * MeSH
• období sucha MeSH
• Poncirus * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Soil arbuscular mycorrhizal fungi (AMF) enhance the tolerance of plants against soil moisture deficit stress (SMDS), but the underlying mechanisms are still not fully understood. Polyamines (PAs) as low-molecular-weight, aliphatic polycations have strong roles in abiotic stress tolerance of plants. We aimed to investigate the effect of AMF (Funneliformis mosseae) inoculation on PAs, PA precursors, activities of PA synthases and degrading enzymes, and concentration of reactive oxygen species in the roots of trifoliate orange (Poncirus trifoliata) subjected to 15 days of SMDS. Leaf water potential and total chlorophyll levels were comparatively higher in AMF-inoculated than in non-AMF-treated plants exposed to SMDS. Mycorrhizal plants recorded a significantly higher concentration of precursors of PA synthesis such as L-ornithine, agmatine, and S-adenosyl methionine, besides higher putrescine and cadaverine and lower spermidine during the 15 days of SMDS. AMF colonization raised the PA synthase (arginine decarboxylase, ornithine decarboxylase, spermidine synthase, and spermine synthase) activities and PA-degrading enzymes (copper-containing diamine oxidase and FAD-containing polyamine oxidase) in response to SMDS. However, mycorrhizal plants showed a relatively lower degree of membrane lipid peroxidation, superoxide anion free radical, and hydrogen peroxide than non-mycorrhizal plants, whereas the difference between them increased linearly up to 15 days of SMDS. Our study concluded that AMF regulated PA homeostasis in roots of trifoliate orange to tolerate SMDS.

• Klíčová slova
• Poncirus trifoliata, citrus, mycorrhiza, polyamine, water deficit,
• Publikační typ
• časopisecké články MeSH

Arbuscular mycorrhizas absorb water from soil to host plants, while the relationship between mycorrhizas and aquaporins (AQPs, membrane water channel proteins, which function in water transport) in mycorrhizal plants is unclear. In this study, Funneliformis mosseae-colonized trifoliate orange (Poncirus trifoliata) seedlings were grown in pots fitted with 37-μm nylon meshes at the bottom of each pot to allow mycorrhizal hyphae absorb water from an outer beaker. The expression of seven plasma membrane intrinsic proteins (PIPs) genes, six tonoplast intrinsic proteins (TIPs) genes, and four nodulin-26 like intrinsic proteins (NIPs) genes were analyzed in roots of both well-watered (WW) and drought stressed (DS) plants. The six-week DS plants dramatically increased hyphal water absorption rate by 1.4 times, as compared with WW plants. Mycorrhizal plants exhibited greater plant growth performance, leaf water status (water potential and relative water content), and gas exchange under both WW and DS conditions. Mycorrhizal inoculation induced diverse expression patterns in these AQPs under WW: up-regulation of PtNIP1;1, PtPIP2;1, and PtPIP2;5, down-regulation of PtNIP1;2, PtNIP6;1, PtPIP1;2, PtPIP1;5, PtPIP2;8, PtTIP1;1, PtTIP1;2, PtTIP1;3, and PtTIP5;1, and no changes in other AQPs. However, the expression of PtPIPs and PtNIPs was down-regulated by mycorrhizal inoculation under DS, and PtTIPs was not induced by mycorrhizal colonization under DS. The expression pattern of AQPs in response to mycorrhizas under DS is a way of mycorrhizal plants to minimize water loss.

• Klíčová slova
• Aquaporins, Hyphal length, Mycorrhization, Soil drought, Trifoliate orange,
• MeSH
• mykorhiza fyziologie   MeSH
• období sucha * MeSH
• Poncirus metabolismus   mikrobiologie   MeSH
• půdní mikrobiologie MeSH
• symbióza MeSH
• Publikační typ
• časopisecké články MeSH

Aquaporins (AQPs) involved in water and small molecule transport respond to environmental stress, while it is not clear how arbuscular mycorrhizal fungi (AMF) regulate AQP expression. Here, we investigated the change in leaf water potential and expression level of four tonoplast intrinsic proteins (TIPs), six plasma membrane intrinsic proteins (PIPs), and four nodin-26 like intrinsic proteins (NIPs) genes in trifoliate orange (Poncirus trifoliata) inoculated with Funneliformis mosseae under well-watered (WW), salt stress (SS), and waterlogging stress (WS). Root AMF colonization and soil hyphal length collectively were reduced by SS and WS. Under WW, inoculation with AMF gave diverse responses of AQPs: six AQPs up-regulated, three AQPs down-regulated, and five AQPs did not change. Such up-regulation of more AQPs under mycorrhization and WW partly accelerated water absorption, thereby, maintaining higher leaf water potential. However, under SS, all the fourteen AQPs were dramatically induced by AMF inoculation, which improved water permeability of membranes and stimulated water transport of the host. Under WS, AMF colonization almost did not induce or even down-regulated these AQPs expressions with three exceptions (PtTIP2;2, PtPIP1;1, and PtNIP1;2), thus, no change in leaf water potential. As a result, mycorrhizal plants under flooding may have an escape mechanism to reduce water absorption. It is concluded that AMF had different strategies in response to environmental stresses (e.g. SS and WS) by regulating leaf AQP expression in the host (e.g. trifoliate orange).

• Klíčová slova
• Abiotic stress, Aquaporin, Citrus, Symbiosis, Water stress,
• MeSH
• akvaporiny * MeSH
• houby MeSH
• listy rostlin metabolismus   MeSH
• mykorhiza * metabolismus   MeSH
• Poncirus * metabolismus   MeSH
• solný stres MeSH
• voda metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• akvaporiny * MeSH
• voda MeSH

The circadian rhythm of plants is associated with stress responses; however, it is not clear whether increased host plant drought tolerance by arbuscular mycorrhizal fungi (AMF) is associated with changes in the circadian clock. The present study aimed to analyze the effect of Funneliformis mosseae (Nicol. & Gerd.) Schüßler & Walker on the circadian clock gene expression patterns in trifoliate orange (Poncirus trifoliata L. Raf.) along with gas exchange, abscisic acid (ABA) levels and antioxidant enzyme gene expression under well-watered (WW) and drought stress (DS) conditions. Plant growth, net photosynthetic rate, stomatal conductance and ABA levels were significantly higher in AMF- than in non-AMF-inoculated plants regardless of soil water regimes. Six circadian clock genes, including PtPRR7, PtLHY, PtCCA1, PtGI, PtPIF3 and PtSRR1, were identified and showed rhythmic expression patterns over the course of the day. The AMF inoculation reduced the expression of most circadian clock genes in different time periods. However, AMF treatment significantly increased PtPRR7 and PtGI expression at 5:00 p.m. under WW and DS conditions, PtLHY expression at 1:00 a.m. and PtSRR1 expression at 9:00 p.m. At 1:00 a.m., AMF inoculation up-regulated the expression of the circadian clock genes PtPRR7, PtCCA1, PtLHY and PtPIF3 and the antioxidant enzyme genes PtFe-SOD, PtMn-SOD, PtCu/Zn-SOD, PtPOD and PtCAT1. Correlation analysis revealed that these changes in circadian clock gene expression were associated with antioxidant enzyme gene expression, root ABA and gas exchange. We concluded that mycorrhizal fungi have the ability to regulate the daily rhythm of the circadian clock in trifoliate orange plants in response to drought.

• Klíčová slova
• arbuscular mycorrhiza, citrus, symbiosis,
• MeSH
• cirkadiánní hodiny * MeSH
• Citrus * MeSH
• mykorhiza * fyziologie   MeSH
• období sucha MeSH
• Poncirus * genetika   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Root hairs and arbuscular mycorrhiza (AM) coexist in root systems for nutrient and water absorption, but the relation between AM and root hairs is poorly known. A pot study was performed to evaluate the effects of four different AM fungi (AMF), namely, Claroideoglomus etunicatum, Diversispora versiformis, Funneliformis mosseae, and Rhizophagus intraradices on root hair development in trifoliate orange (Poncirus trifoliata) seedlings grown in sand. Mycorrhizal seedlings showed significantly higher root hair density than non-mycorrhizal seedlings, irrespective of AMF species. AMF inoculation generally significantly decreased root hair length in the first- and second-order lateral roots but increased it in the third- and fourth-order lateral roots. AMF colonization induced diverse responses in root hair diameter of different order lateral roots. Considerably greater concentrations of phosphorus (P), nitric oxide (NO), glucose, sucrose, indole-3-acetic acid (IAA), and methyl jasmonate (MeJA) were found in roots of AM seedlings than in non-AM seedlings. Levels of P, NO, carbohydrates, IAA, and MeJA in roots were correlated with AM formation and root hair development. These results suggest that AMF could alter the profile of root hairs in trifoliate orange through modulation of physiological activities. F. mosseae, which had the greatest positive effects, could represent an efficient AM fungus for increasing fruit yields or decreasing fertilizer inputs in citrus production.

• Klíčová slova
• Citrus, Glucose, IAA, MeJA, Mycorrhiza, Root hairs,
• MeSH
• biomasa MeSH
• Citrus růst a vývoj   mikrobiologie   MeSH
• Glomeromycota fyziologie   MeSH
• kořeny rostlin růst a vývoj   metabolismus   mikrobiologie   MeSH
• mykorhiza růst a vývoj   fyziologie   MeSH
• Poncirus růst a vývoj   mikrobiologie   MeSH
• půda MeSH
• půdní mikrobiologie MeSH
• semenáček růst a vývoj   metabolismus   mikrobiologie   MeSH
• symbióza fyziologie   MeSH
• výhonky rostlin růst a vývoj   metabolismus   mikrobiologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• půda MeSH

Arbuscular mycorrhizal fungi (AMF) can enhance drought tolerance in plants, whereas little is known regarding AMF contribution to sucrose and proline metabolisms under drought stress (DS). In this study, Funneliformis mosseae and Paraglomus occultum were inoculated into trifoliate orange (Poncirus trifoliata) under well watered and DS. Although the 71-days DS notably (P < 0.05) inhibited mycorrhizal colonization, AMF seedlings showed significantly (P < 0.05) higher plant growth performance and leaf relative water content, regardless of soil water status. AMF inoculation significantly (P < 0.05) increased leaf sucrose, glucose and fructose concentration under DS, accompanied with a significant increase of leaf sucrose phosphate synthase, neutral invertase, and net activity of sucrose-metabolized enzymes and a decrease in leaf acid invertase and sucrose synthase activity. AMF inoculation produced no change in leaf ornithine-δ-aminotransferase activity, but significantly (P < 0.05) increased leaf proline dehydrogenase activity and significantly (P < 0.05) decreased leaf both Δ1-pyrroline-5-carboxylate reductase and Δ1-pyrroline-5-carboxylate synthetase activity, resulting in lower proline accumulation in AMF plants under DS. Our results therefore suggest that AMF strongly altered leaf sucrose and proline metabolism through regulating sucrose- and proline-metabolized enzyme activities, which is important for osmotic adjustment of the host plant.

• MeSH
• fyziologický stres * MeSH
• Glomeromycota fyziologie   MeSH
• listy rostlin enzymologie   metabolismus   MeSH
• metabolismus sacharidů MeSH
• mykorhiza fyziologie   MeSH
• období sucha * MeSH
• počet mikrobiálních kolonií MeSH
• Poncirus růst a vývoj   metabolismus   mikrobiologie   MeSH
• prolin metabolismus   MeSH
• sacharosa metabolismus   MeSH
• semenáček metabolismus   MeSH
• voda metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• prolin MeSH
• sacharosa MeSH
• voda MeSH

Root-hair growth and development regulated by soil microbes is associated with auxin. In this background, we hypothesized that mycorrhizal fungal inoculation induces greater root-hair growth through stimulated auxin synthesis and transport under water stress conditions. Trifoliate orange (Poncirus trifoliata) was inoculated with an arbuscular mycorrhizal (AM) fungus (Funneliformis mosseae) under well-watered (WW) and drought stress (DS) for 9 weeks. Compared with non-AM seedlings, AM seedlings displayed significantly higher density, length, and diameter of root hairs and root indoleacetic acid (IAA) level, whereas lower total root IAA efflux, regardless of soil moisture status. Root PtYUC3 and PtYUC8 involved in IAA biosynthesis were up-regulated by mycorrhization under WW and DS, whereas AM-modulated expression in PtTAA1, PtTAR2, PtYUC4, and PtYUC6 depended on status of soil moisture. Mycorrhizal inoculation down-regulated the transcript level of root auxin efflux carriers like PtPIN1 and PtPIN3, whereas significantly up-regulated the expression of root auxin-species influx carriers like PtABCB19 and PtLAX2 under DS. These results indicated that AMF-stimulated greater root-hair growth of trifoliate orange under DS that is independent on AMF species is related with mycorrhiza-modulated auxin synthesis and transport, which benefits the host plant to enhance drought tolerance.

• MeSH
• biologický transport MeSH
• fyziologický stres * MeSH
• Glomeromycota růst a vývoj   fyziologie   MeSH
• kyseliny indoloctové metabolismus   MeSH
• messenger RNA genetika   metabolismus   MeSH
• mykorhiza růst a vývoj   fyziologie   MeSH
• období sucha * MeSH
• počet mikrobiálních kolonií MeSH
• Poncirus genetika   růst a vývoj   mikrobiologie   fyziologie   MeSH
• regulace genové exprese u rostlin MeSH
• rostlinné geny MeSH
• rostlinné proteiny genetika   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• indoleacetic acid MeSH Prohlížeč
• kyseliny indoloctové MeSH
• messenger RNA MeSH
• rostlinné proteiny MeSH