Mycorrhizal colonization with Glomus intraradices and development stage of transformed tomato roots significantly modify the chemotactic response of zoospores of the pathogen Phytophthora nicotianae

The chemotaxic response of zoospores of the plant pathogen, Phytophthora nicotianae, towards exudates from mycorrhizal and non-mycorrhizal transformed tomato roots was studied. A bi-compartmental in vitro system was used to grow Ri T-DNA-transformed tomato roots colonized or non-colonized with the arbuscular mycorrhizal fungus, Glomus intraradices, and to collect root and mycorrhizal exudates. The root and mycorrhizal growth dynamics were first characterized in order to determine two times of exudate sampling. Exudates collected from 16-wk-old mycorrhizal roots were significantly more attractive for P. nicotianae zoospores than exudates from non-inoculated roots. On the contrary, concentrated exudates harvested from 24-wk-old mycorrhizal roots were repulsive to zoospores compared to exudates from non-colonized roots and the water control. In exudates of G. intraradices-inoculated roots, HPLC–MS analyses revealed significantly higher concentrations of proline and isocitrate after 24 wk of growth, while after 16 wk of growth, proline concentration did not differ between exudate types, and the isocitrate concentration was lower in mycorrhizal root exudates. Mycorrhizal inoculation had no effect on the amounts of other amino acids and organic acids and on the sugars quantified within exudates. Our results suggest that modification in exudate composition of mature roots by mycorrhizal colonization may provoke the repulsion of P. nicotianae, and that their capacity to infect host roots may in this manner be reduced.     

The bacterial community of tomato rhizosphere is modified by inoculation with arbuscular mycorrhizal fungi but unaffected by soil enrichment with mycorrhizal root exudates or inoculation with Phytophthora nicotianae

Arbuscular mycorrhizal (AM) fungi have been shown to induce the biocontrol of soilborne diseases, to change the composition of root exudates and to modify the bacterial community structure of the rhizosphere, leading to the formation of the mycorrhizosphere. Tomato plants were grown in a compartmentalized soil system and were either submitted to direct mycorrhizal colonization or to enrichment of the soil with exudates collected from mycorrhizal tomato plants, with the corresponding negative controls. Three weeks after planting, the plants were inoculated or not with the soilborne pathogen Phytophthora nicotianae growing through a membrane from an adjacent infected compartment. At harvest, a PCR-Denaturing gradient gel electrophoresis analysis of 16S rRNA gene fragments amplified from the total DNA extracted from each plant rhizosphere was performed. Root colonization with the AM fungi Glomus intraradices or Glomus mosseae induced significant changes in the bacterial community structure of tomato rhizosphere, compared to non-mycorrhizal plants, while enrichment with root exudates collected from mycorrhizal or non-mycorrhizal plants had no effect. Our results support that the effect of AM fungi on rhizosphere bacteria would not be mediated by compounds present in root exudates of mycorrhizal plants but rather by physical or chemical factors associated with the mycelium, volatiles and/or root surface bound substrates. Moreover, infection of mycorrhizal or non-mycorrhizal plants with P. nicotianae did not significantly affect the bacterial community structure suggesting that rhizosphere bacteria would be less sensitive to the pathogen invasion than to mycorrhizal colonization. Of 96 unique sequences detected in the tomato rhizosphere, eight were specific to mycorrhizal fungi, including two Pseudomonas, a Bacillus simplex, an Herbaspirilium and an Acidobacterium. One Verrucomicrobium was common to rhizospheres of mycorrhizal plants and of plants watered with mycorrhizal root exudates.     

丛枝菌根与植物寄生性线虫相互作用及抗性机制

丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)能够与大多数陆地植物互惠共生,促进植物对养分的吸收,提高植物对各种生物和非生物胁迫的抗逆性,对植物健康生长有重要的作用。在土壤中丛枝菌根真菌与植物寄生性线虫共同依靠寄主植物根系完成生命循环,但二者对寄主植物作用完全相反,引起研究者广泛兴趣,成为菌根研究的热点和焦点之一。本文分析了丛植菌根真菌与植物寄生线虫的相互作用,并探讨了菌根提高植物对线虫抗性的可能机制:菌根真菌改善植物的生长和营养状况、改变植物根系形态结构、影响根系分泌物和根际微生物区系、诱导寄主植物产生防御反应等,旨在深入挖掘丛枝菌根真菌的生物学功能,进一步发挥其在农业生产中的应用潜力。     

Soil sickness of peanuts is attributable to modifications in soil microbes induced by peanut root exudates rather than to direct allelopathy

The quantity and quality of peanut yields are seriously compromised by consecutive monoculture in the subtropical regions of China. Root exudates, which represent a growth regulator in peanut–soil feedback processes, play a principal role in soil sickness. The growth inhibition of a species in an in vitro bioassay enriched with root exudates and allelochemicals is commonly viewed as evidence of an allelopathic interaction. However, for some of these putative examples of allelopathy, the results have not been verified in more natural settings with plants continuously growing in soil. In this study, the phenolic acids in peanut root exudates, their retention characteristics in an Udic Ferrosol, and their effects on rhizosphere soil microbial communities and peanut seedling growth were studied. Phenolic acids from peanut root exudates were quickly metabolized by soil microorganisms and did not accumulate to high levels. The peanut root exudates selectively inhibited or stimulated certain communal bacterial and fungal species, with decreases in the relative abundance of the bacterial taxa Gelria glutamica, Mitsuaria chitosanitabida, and Burkholderia soli and the fungal taxa Mortierella sp. and Geminibasidium hirsutum and increases in the relative abundance of the bacterial taxon Desulfotomaculum ruminis and the fungal taxa Fusarium oxysporum, Bionectria ochroleuca and Phoma macrostoma. The experimental application of phenolic acids to non-sterile and sterile soil revealed that the poor performance of the peanut plants was attributed to changes in the soil microbial communities promoted by phenolic acids. These results suggest that pathogenic fungal accumulation at the expense of such beneficial microorganisms as plant growth promoting rhizobacteria, mycorrhizal fungi induced by root exudates, rather than direct autotoxicity induced by root exudates, might represent the principal cause underlying the soil sickness associated with peanut plants. We hope that our study will motivate researchers to integrate the role of soil microbial communities in allelopathic research, such that their observed significance in soil sickness during continuous monocropping of fields can be further explored.     

Pre-symbiotic and symbiotic interactions between Glomus intraradices and two Paenibacillus species isolated from AM propagules. In vitro and in vivo assays with soybean (AG043RG) as plant host

Two indole-producing Paenibacillus species, known to be associated with propagules of arbuscular mycorrhizal (AM) fungi, were examined for their mycorrhization helper bacteria activity at pre-symbiotic and symbiotic stages of the AM association. The effects were tested under in vitro and in vivo conditions using an axenically propagated strain of the AM fungus Glomus intraradices and Glycine max (soybean) as the plant host. The rates of spore germination and re-growth of intraradical mycelium were not affected by inoculation with Paenibacillus strains in spite of the variation of indole production measured in the bacterial supernatants. However, a significant promotion in pre-symbiotic mycelium development occurred after inoculation of both bacteria under in vitro conditions. The Paenibacillus rhizosphaerae strain TGX5E significantly increased the extraradical mycelium network, the rates of sporulation, and root colonization in the in vitro symbiotic association. These results were also observed in the rhizosphere of soybean plants grown under greenhouse conditions, when P. rhizosphaerae was co-inoculated with G. intraradices. However, soybean dry biomass production was not associated with the increased development and infectivity values of G. intraradices. Paenibacillus favisporus strain TG1R2 caused suppression of the parameters evaluated for G. intraradices during in vitro symbiotic stages, but not under in vivo conditions. The extraradical mycelium network produced and the colonization of soybean roots by G. intraradices were promoted compared to the control treatments. In addition, dual inoculation had a promoting effect on soybean biomass production. In summary, species of Paenibacillus associated with AM fungus structures in the soil, may have a promoting effect on short term pre-symbiotic mycelium development, and little impact on AM propagule germination. These findings could explain the associations found between some bacterial strains and AM fungus propagules.     

Screening Musa germplasm for resistance to burrowing nematode populations from Uganda

Nematode pathogenicity experiments on Musa host plants were carried out in pot trials. Both the final nematode population densities and percentages root necrosis on different host plants were higher for the Radopholus similis population from Mbarara than for the populations from Namulonge, Ikulwe and Mukono (Uganda). The R. similis population from Mbarara managed to break the resistance of the diploid banana Pisang Jari Buaya, a known worldwide source of resistance to R. similis. The diploid banana hybrid TMB2× 9128-3 and the triploid dessert banana Yangambi km5 showed resistance against the four R. similis populations. These results indicate that differences in pathogenicity among different R. similis populations exist and should be taken into consideration for plantain and banana germplasm enhancement.     

Improved zinc tolerance in Eucalyptus globulus inoculated with Glomus deserticola and Trametes versicolor or Coriolopsis rigida

The potential of interactions between saprophytic and arbuscular mycorrhizal (AM) fungi to improve Eucalyptus globulus grown in soil contaminated with Zn were investigated. The presence of 100 mg kg ⁻¹ Zn decreased the shoot and root dry weight of E. globulus colonized with Glomus deserticola less than in plants not colonized with AM. Zn also decreased the extent of root length colonization by AM and the AM fungus metabolic activity, measured as succinate dehydrogenase (SDH) activity of the fungal mycelium inside the E. globulus root. The saprophytic fungi Trametes versicolor and Coriolopsis rigida increased the shoot dry weight and the tolerance of E. globulus to Zn when these plants were AM-colonized. Both saprophytic fungi increased the percentage of AM root length colonization and elevated G. deserticola SDH activity in the presence of all Zn concentrations applied to the soil. In the presence of 500 and 1000 mg kg⁻¹ Zn, there were higher metal concentrations in roots and shoots of AM than in non-AM plants; furthermore, both saprophytic fungi increased Zn uptake by E. globulus colonized by G. deserticola. The higher root to shoot metal ratio observed in mycorrhizal E. globulus plants indicates that G. deserticola enhanced Zn uptake and accumulation in the root system, playing a filtering/sequestering role in the presence of Zn. However, saprophytic fungi did not increase the root to shoot Zn ratio in mycorrhizal E. globulus plants. The effect of the saprophytic fungi on the tolerance and the accumulation of Zn in E. globulus was mediated by its effect on the colonization and metabolic activity of the AM fungi.     

Root herbivory by Tipula paludosa larvae increases colonization of Agrostis capillaris by arbuscular mycorrhizal fungi

The objective of this study was to determine if root-feeding by insect larvae affects subsequent colonization of roots by arbuscular mycorrhizal fungi. We investigated grazing by larvae of Tipula paludosa on colonization of Agrostis capillaris by two species of fungi, Glomus mosseae and G. intraradices. Host plants were subjected to 7 days of grazing only, continuous grazing for 42 days, or no herbivory. Those plants with no herbivory had significantly lower levels of colonization by arbuscules and hyphae compared to plants which were grazed for 7 or 42 days. The effect only occurred in the upper parts of the root system, where larvae were active. We suggest that this effect was most likely mediated by a change in quantity and composition of root exudates.     

Root exudates as pre-invasive factors in the modulation of chick pea varieties

Five chick pea (Cicer arietinum L.) varieties when inoculated with an effective Rhizobium strain showed significant variations in nodule number. The root exudates of the plant varieties were analysed for sugars, TCA cycle intermediates and amino acids. Xylose, citrate, alanine and phenylalanine were detected in exudates of one or two varieties, in addition to compounds common to all varieties such as glucose, glucosamine, ribose, serine, homoserine and glutamic acid. However, the concentration of all these compounds in the exudates collectively was inadequate to support Rhizobium growth under aseptic conditions. Several of the exuded compounds were chemotactic at very low concentrations. The rhizobia showed taxes toward growing roots of all the varieties in agar plates. Competition experiments showed that the rhizobia have a multiple-chemotactic system like that of Escherichia coli. It appears that root exudates accumulate Rhizobium along the growing roots through chemotaxis. The minor differences in exudate composition among varieties is not a factor responsible for variation in nodule number.     

Potential of seed and root exudates of the common bean Phaseolus vulgaris L. for immediate induction of rhizobial chemotaxis and nod genes

Abstract

Immediate induction of rhizobial chemotaxis and nod genes by seed and root exudates of the common bean Phaseolus vulgaris L. was investigated. One hour after the onset of Rhizobium incubation in a culture medium composed of common bean exudates and soil extract, a large number of Rhizobium cells were attracted by the seed exudates of all the cultivars (cvs Hokkaikintoki, Himetebou and Kurodanekinugasa), while the media containing root exudates was not statistically different from the control. After 1 h of incubation, the nod genes were induced significantly by the seed exudates of Himetebou and Kurodanekinugasa. In contrast, the seed exudate of Hokkaikintoki and the root exudates of all the cultivars failed to exert a beneficial effect on the immediate induction of the nod genes. These results suggested that the seed exudates displayed a higher potential for immediate induction of rhizobial chemotaxis and nod genes than the root exudates, except for nod gene induction by the seed exudate of Hokkaikintoki.     

Chemotaxis of deleterious rhizobacteria to birdsfoot trefoil

Intact seeds and seed and seedling root exudates of birdsfoot trefoil (Lotus corniculatus L.) were used as chemoattractants in experiments to determine the relative importance of chemotaxis in spermosphere and rhizosphere colonization by selected rhizobacteria. Results for soft-agar, capillary tube and soil chemotaxis assays indicated that selected deleterious rhizobacteria were attracted to seed and seedling root exudates. Several sugars and phenolic fractions detected in exudates were chemoattractants for these rhizobacteria. Using soil-chemotaxis assemblies, migration of rhizobacterial isolates through 2 cm distances of soil toward birdsfoot trefoil seeds was detected within 24 h. Isolates were not detected at the same site in soils without seeds until 72 h after inoculation. These results suggest that attraction of deleterious rhizobacteria toward seeds and seedling roots mediated by exudates (chemotaxis) might be the first step in the establishment and subsequent colonization of bacteria involved in soilborne disease complexes of birdsfoot trefoil.     

Take-all disease is systemically reduced in roots of mycorrhizal barley plants

The systemic effect of root colonization by the arbuscular mycorrhizal fungus Glomus mosseae on infection of barley by Gaeumannomyces graminis var. tritici (Ggt) was studied. In split-root systems of barley one side was inoculated with G. mosseae and the other side was inoculated with Ggt.Root infection by Ggt was systemically reduced when barley plants showed high degrees of mycorrhizal root colonization, whereas a low mycorrhizal root colonization exhibited no effect on Ggt infection. Our results show a clear systemic bioprotectional effect depending on the degree of root colonization by the mycorrhizal fungus. At a higher mycorrhizal colonization rate the concentration of salicylic acid (SA) was increased in roots colonized by the mycorrhizal fungus but no systemic increase of SA could be measured in non-mycorrhizal roots of mycorrhizal plants, indicating that the systemic bioprotectional effect against Ggt is not mediated by salicylic acid.     

Nematode dispersion by runoff water: Case study of Radopholus similis (Cobb) Thorne on nitisol under humid tropical conditions

To minimize application of nematicides in banana fields, crop systems have been developed in the French West Indies that combine fallow or rotation crops and nematode-free in vitro plants. After two to four years, populations of the burrowing nematode Radopholus similis have developed enough to cause economic losses, leading banana growers to use nematicides. To understand how banana fields are recontaminated, we studied the dissemination of R. similis by water flow. At a 1-m scale, we analyzed the dispersion of R. similis under a rainfall simulator: we isolated a 1-m² study plot, placed a R. similis suspension on the upstream soil surface, and simulated a 60 mm/h rainfall for 72 min. We collected soil samples every 10 cm downstream after 12 min of rainfall, and subsequently at 20-min intervals, and extracted the nematodes using a Seinhorst elutriator and then a Baermann funnel. Our results showed that the nematode dissemination follows an inverse exponential law, and depends more on soil moisture at the beginning of rainfall than on the length of rainfall: in fresh soil, 69–80% of the R. similis recovered were found less than 10 cm downstream from the nematode inoculation line, whereas in wetted soil, 76–85% of the recovered individuals were collected in the outlet tub located downstream from the apparatus. This passive dissemination model partially explains the distance covered by individual nematodes but not the low percentage of motile nematodes recovered in the outlet tub (10% and 36% in fresh and wet soils) compared to the percentage of motile nematodes found in the soil (80% and 84% in fresh and wet soils). Indeed, water runoff is likely to disseminate R. similis over long distances only when soil moisture is close to field capacity.     

Effects of an arbuscular mycorrhizal fungus and two plant-parasitic nematodes on<Emphasis Type="Italic"> Musa</Emphasis> genotypes differing in root morphology

In this study, the effect of an arbuscular mycorrhizal fungus (AMF) and two migratory endoparasitic nematodes on Musa plant growth, including the root system, were examined. In addition, the AMF-nematode interaction was studied. Seven Musa genotypes with different root systems were selected. Based on their relative mycorrhizal dependency, two genotypes (Calcutta 4 and Obino l'Ewai) were selected for AMF-nematode interaction studies. The experiments were performed under greenhouse conditions. Mycorrhization with Glomus mosseae resulted in a significantly better plant growth even in the presence of nematodes. The effect of AMF on the root system was genotype-dependent and seemed to be related to the relative mycorrhizal dependency of the genotype. The nematodes also affected the root system, decreasing branching. Nematode population densities were significantly reduced in the presence of AMF, except for Pratylenchus coffeae in Obino l'Ewai. In the root system, it appeared that the decreased branching caused by the nematodes was counterbalanced by the increased branching caused by the AMF.     

Role of wheat root exudates in associative nitrogen fixation

Root exudates and related exudate diffusion gradients were studied using ¹⁴C-radioisotope techniques. With inoculated wheat plants (Triticum aestivum cv. Nugaines), 3.7% of the ¹⁴C-labeled photosynthate was released as soluble exudate whereas 3.0% was found with axenic plants. Root surface areas averaged 54 cm² plant^?1. The microbial cells produced were sufficient to colonize 7.4%, of the root surface with a cell monolayer. Gradient studies showed that with inoculated root systems, rapid utilization of soluble exudate markedly decreased the distance of exudate diffusion. Microbial colonization also was a function of the physiological features of the test culture. The relationship between root colonization, exudate production and potential for associative nitrogen fixation is discussed.     

Water status and stomatal behaviour of cowpea,Vigna unguiculata (L.) Walp,plants inoculated with two Glomus species at low soil moisture levels

The effects of arbuscular mycorrhizal (AM) fungi on water status and stomatal behaviour of cowpea, Vigna unguiculata (L.) Walp. cv. B89-504, under water-stressed conditions in the greenhouse were studied. The 3 × 2 experimental design included two levels of mycorrhizal colonisation (Glomus mosseae, Glomus versiforme) and non-mycorrhizal control treatment and two soil moisture levels (well-watered pots and pots allowed to dry). Relative water content and leaf water potential values were higher in well-watered mycorrhizal and non-mycorrhizal plants than in water-stressed mycorrhizal and non-mycorrhizal plants. AM species had no significant effect on leaf osmotic potential, stomatal conductance and leaf transpiration in both well watered and water-stressed plants. The values of stomatal conductance and leaf transpiration were high during the vegetative stage and low during the flowering stage. These responses which can be related to the age of the plant suggest that mycorrhizal colonisation did not affect stomatal closure of cowpea plants during water stress. The decrease in plant growth and dry matter production in both mycorrhizal and non-mycorrhizal plants shows that drought resistance in cowpea was unaffected by mycorrhiza in the vegetative phase.     

Influence of the sunflower rhizosphere on the biodegradation of PAHs in soil

Reduced bioavailability to soil microorganisms is probably the most limiting factor in the bioremediation of polycyclic aromatic hydrocarbons PAH-polluted soils. We used sunflowers planted in pots containing soil to determine the influence of the rhizosphere on the ability of soil microbiota to reduce PAH levels. The concentration of total PAHs decreased by 93% in 90 days when the contaminated soil was cultivated with sunflowers, representing an improvement of 16% compared to contaminated soil without plants. This greater extent of PAH degradation was consistent with the positive effect of the rhizosphere in selectively stimulating the growth of PAH-degrading populations. Molecular analysis revealed that the increase in the number of degraders was accompanied by a dramatic shift in the structure of the bacterial soil community favoring groups with a well-known PAH-degrading capacity, such as Sphingomonas (α-Proteobacteria), Commamonas and Oxalobacteria (β-Proteobacteria), and Xhanthomonas (γ-Proteobacteria). Other groups that were promoted for which degrading activity has not been reported included Methylophyllus (β-Proteobacteria) and the recently described phyla Acidobacteria and Gemmatimonadetes. We also conducted mineralization experiments on creosote-polluted soil in the presence and absence of sunflower root exudates to advance our understanding of the ability of these exudates to serve as bio-stimulants in the degradation of PAHs. By conducting greenhouse and mineralization experiments, we separated the chemical impact of the root exudates from any root surface phenomena, as sorption of contaminants to the roots, indicating that sunflower root exudates have the potential to increase the degradation of xenobiotics due to its influence on the soil microorganisms, where sunflower root exudates act improving the availability of the contaminant to be degraded. We characterized the sunflower exudates in vitro to determine the total organic carbon (TOC) and its chemical composition. Our results indicate that the rhizosphere promotes the degradation of PAHs by increasing the biodegradation of the pollutants and the number and diversity of PAH degraders. We propose that the biostimulation exerted by the plants is based on the chemical composition of the exudates.     

Interactions between arbuscular mycorrhizal fungi and fungivorous nematodes on the growth and arsenic uptake of tobacco in arsenic-contaminated soils

The effects of inoculation with two AM fungi (M1, Glomus caledonium; M2, Glomus spp. and Acaulospora spp.) and a fungivorous nematode Aphelenchoides sp. on growth and arsenic (As) uptake of Nicotiana tabacum L. were investigated in soils contaminated with a range of As. The reproduction of Aphelenchoides sp. was triggered by the co-inoculation of AM fungi regardless of AM fungal isolates and As levels. Stimulative effects of Aphelenchoides sp. on the development of mycorrhiza, slightly different between two AM fungi, were found particularly at the lowest As level. Irrespective of mycorrhizal inoculi, increasing soil As level decreased plant growth, but increased plant As uptake. Co-inoculation of AM fungi and Aphelenchoides sp. led plants to achieving further growth and greater As accumulation at the lowest As level. Results showed that the interactions between AM fungi and fungivorous nematodes were important in plant As tolerance and phytoextraction at low level As-polluted soil.     

Fluorescence detection of aluminum in arbuscular mycorrhizal fungal structures and glomalin using confocal laser scanning microscopy

Arbuscular mycorrhizal spores and glomalin-related soil protein (GRSP) isolated from acid soils were analyzed using confocal laser scanning microscopy (CLSM) for Al detection. Mycorrhizal structures of Glomus intraradices produced under in vitro conditions as well as spores and GRSP from neutral and Cu-polluted soils were used as contrasting criteria. Spores and GRSP from soils with 7 and 70% Al saturation showed autofluorescence which increased especially at the highest soil Al level and when Al³⁺ solution was added. G. intraradices spores showed fluorescence only when exogenous Al³⁺ was added. On the contrary, spores and GRSP from neutral and Cu-polluted soils showed little or no significant fluorescence. This fluorescence shown by fungal structures and GRSP when subjected to high Al (of endogenous or exogenous origin) suggest a high capacity for Al immobilization, which could be an effective way to reduce Al activity and phytotoxicity in acid soils.     

Root exudate-stimulated RNA accumulation in the arbuscular mycorrhizal fungus Gigaspora rosea

Plant root exudates induce the transition from asymbiosis to presymbiosis in arbuscular mycorrhizal fungi. In order to get an insight into this developmental switch, two libraries of Gigaspora rosea and one library of Gigaspora gigantea were screened for fragments of genes that show enhanced RNA accumulation 1 h after addition of a semi-purified exudate fraction of carrot roots. Among 150 clones, 40 seemed to contain inserts of root exudate-induced genes. One of the genes, GrosRbp1, putatively encoding an RNA binding protein involved in developmental control showed RNA accumulation which correlates to the extent of stimulation of presymbiotic hyphal branching.