Arbuscular Mycorrhizal Fungi and Endophytic Fungi Activate Leaf Antioxidant Defense System of Lane Late Navel Orange.

Arbuscular mycorrhizal (AM) fungi and endophytic fungi collectively symbiose well with plants and, thus, stimulate plant growth; however, it is not clear whether field inoculation of the fungi enhances the resistance potential of plants, particularly in citrus. In the present study, we inoculated AM fungi (Acaulospora scrobiculata, Diversispora spurca, and D. versiformis) and endophytic fungi (Piriformospora indica) on an eight-year-old lane late navel orange (Citrus sinensis (L.) Osb) trees grafted on Poncirus trifoliata in a field, and we analyzed the response of the leaf antioxidant defense system. Approximately 2 years after inoculation, the root fungal colonization rate and soil hyphal length significantly increased. Fungal inoculation significantly increased the activity of leaf antioxidant enzymes, such as superoxide dismutase, peroxidase, catalase, and ascorbate peroxidase, and the content of non-enzymatic antioxidants, such as reduced ascorbic acid and reduced glutathione. As a result, fungi-inoculated plants maintained lower concentrations of hydrogen peroxide and superoxide anion radicals and lower levels of membrane lipid peroxidation (according to malondialdehyde level) in leaves than uninoculated plants. Among them, inoculation of D. spurca and A. scrobiculata showed relatively higher effects in enhancing the antioxidant defense system than the other fungi. Furthermore, inoculation of D. spurca induced expressions of CsFe-SOD, CsMn-SOD, CsPOD, CsCAT1, and CsPRR7; inoculation of A. scrobiculata and D. versiformis induced expressions of CsCAT1; CsCAT1 and CsPOD were also induced by inoculation of P. indica. All four inoculations almost upregulated expressions of CsFAD6. AM fungi had superior effects than endophytic fungi (e.g., P. indica). According to our findings, inoculation with beneficial fungi, specifically mycorrhizal fungus D. spurca, activated the antioxidant defense system of field citrus trees, thus, having potentially superior resistance in inoculated plants.

Mycorrhizas alter sucrose and proline metabolism in trifoliate orange exposed to drought stress.

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.

Effects of Common Mycorrhizal Network on Plant Carbohydrates and Soil Properties in Trifoliate Orange-White Clover Association.

Common mycorrhizal network (CMN) allows nutrients and signals to pass between two or more plants. In this study, trifoliate orange (Poncirus trifoliata) and white clover (Trifolium repens) were planted in a two-compartmented rootbox, separated by a 37-µm nylon mesh and then inoculated with an arbuscular mycorrhizal fungus (AMF), Diversispora spurca. Inoculation with D. spurca resulted in formation of a CMN between trifoliate orange and white clover, whilst the best AM colonization occurred in the donor trifoliate orange-receptor white clover association. In the trifoliate orange-white clover association, the mycorrhizal colonization of receptor plant by extraradical hyphae originated from the donor plant significantly increased shoot and root fresh weight and chlorophyll concentration of the receptor plant. Enzymatic activity of soil ß-glucoside hydrolase, protease, acid and neutral phosphatase, water-stable aggregate percentage at 2-4 and 0.5-1 mm size, and mean weight diameter in the rhizosphere of the receptor plant also increased. The hyphae of CMN released more easily-extractable glomalin-related soil protein and total glomalin-related soil protein into the receptor rhizosphere, which represented a significantly positive correlation with aggregate stability. AMF inoculation exhibited diverse changes in leaf and root sucrose concentration in the donor plant, and AM colonization by CMN conferred a significant increase of root glucose in the receptor plant. These results suggested that CMN formed in the trifoliate orange-white clover association, and root AM colonization by CMN promoted plant growth, root glucose accumulation, and rhizospheric soil properties in the receptor plant.