Differential effects of AM fungal isolates on Medicago truncatula growth and metal uptake in a multimetallic (Cd, Zn, Pb) contaminated agricultural soil

Toxic metal accumulation in soils of agricultural interest is a serious problem needing more attention, and investigations on soil–plant metal transfer must be pursued to better understand the processes involved in metal uptake. Arbuscular mycorrhizal (AM) fungi are known to influence metal transfer in plants by increasing plant biomass and reducing metal toxicity to plants even if diverging results were reported. The effects of five AM fungi isolated from metal contaminated or non-contaminated soils on metal (Cd, Zn) uptake by plant and transfer to leachates was assessed with Medicago truncatula grown in a multimetallic contaminated agricultural soil. Fungi isolated from metal-contaminated soils were more effective to reduce shoot Cd concentration. Metal uptake capacity differed between AM fungi and depended on the origin of the isolate. Not only fungal tolerance and ability to reduce metal concentrations in plant but also interactions with rhizobacteria affected heavy metal transfer and plant growth. Indeed, thanks to association with nodulating rhizobacteria, one Glomus intraradices inoculum increased particularly plant biomass which allowed exporting twofold more Cd and Zn in shoots as compared to non-mycorrhizal treatment. Cd concentrations in leachates were variable among fungal treatments, but can be significantly influenced by AM inoculation. The differential strategies of AM fungal colonisation in metal stress conditions are also discussed.     

Influence of arbuscular-mycorrhizal fungi, Rhizobium meliloti strains and PGPR inoculation on the growth of Medicago arborea used as model legume for re-vegetation and biological reactivation in a semi-arid mediterranean area

Medicago arborea can be used for re-vegetationpurposes under semiarid conditions. These woody legumes have the ability toforman association with arbuscular mycorrhizal (AM) fungi and rhizobial bacteria,which can be maximised by microorganisms producing certain stimulatingmetabolites acting as plant growth promoting rhizobacteria (PGPR). The effectsof single and combined inoculations using microorganisms with different andinteractive metabolic capacities, namely three Glomusspecies, two Rhizobium meliloti strains (a wild type, WTand its genetically modified derivative GM) and a plant growth promotingrhizobacterium, (PGPR), were evaluated. All three inoculated AM fungi affectedMedicago growth in different ways. Differences weremaintained when soil was co-inoculated with each of the rhizobial strains (WTorGM) and the PGPR. Mycorrhizal fungi were effective in all cases, but the PGPRonly affected plant growth specific microbial situations. PGPR increased growthof G. mosseae-colonised plants associated withRhizobium WT strain by 36% and those infected byG. deserticola when associated with the rhizobial GMstrainby 40%. The most efficient microbial treatments involved mycorrhizalinoculation, which was an indication of the AM dependency of this plantspecies.Moreover, PGPR inoculation was only effective when associated with specificmycorrhizal endophytes (G. mosseae plus WT andG.deserticola plus GM rhizobial strain). The reduced root/shoot (R/S)ratio resulting from PGPR inoculation, was an indication of more effective rootfunction in treated plants. AM colonisation and nodule formation wereunaffectedby the type of AM fungus or bacteria (rhizobial strain and/or PGPR). AM fromnatural soil were less infective and effective than those from the collection.The results supported the existence of selective microbial interactionsaffecting plant performance. The indigenous AM fungi appeared to be ineffectiveand M. arborea behaved as though it was highly dependentonAM colonisation, which implied that it must have a mycorrhizal association toreach maximum growth in the stressed conditions tested. Optimum growth ofmycorrhizal M. arborea plants was associated with specificmicrobial groups, accounting for a 355% increase in growth overnodulatedcontrol plants. The beneficial effect of PGPR in increasing the growth of awoody legume, such as M. arborea under stress, was onlyobserved with co-inoculation of specific AM endophytes. As a result of theinteraction, only shoot biomass was enhanced, but not as a consequence ofenhancing of the colonising abilities of the endophytes. The growthstimulation,occurring as a consequence of selected microbial groups, may be critical anddecisive for the successful establishment of plants under Mediterraneanclimaticand soil conditions.     

Interactions between arbuscular mycorrhizal fungi and soil bacteria

The soil environment is interesting and complicated. There are so many interactions taking place in the soil, which determine the properties of soil as a medium for the growth and activities of plants and soil microorganisms. The soil fungi, arbuscular mycorrhiza (AM), are in mutual and beneficial symbiosis with most of the terrestrial plants. AM fungi are continuously interactive with a wide range of soil microorganisms including nonbacterial soil microorganisms, plant growth promoting rhizobacteria, mycorrhiza helper bacteria and deleterious bacteria. Their interactions can have important implications in agriculture. There are some interesting interactions between the AM fungi and soil bacteria including the binding of soil bacteria to the fungal spore, the injection of molecules by bacteria into the fungal spore, the production of volatiles by bacteria and the degradation of fungal cellular wall. Such mechanisms can affect the expression of genes in AM fungi and hence their performance and ecosystem productivity. Hence, consideration of such interactive behavior is of significance. In this review, some of the most important findings regarding the interactions between AM fungi and soil bacteria with some new insights for future research are presented.     

Trade-offs between arbuscular mycorrhizal fungal competitive ability and host growth promotion in Plantago lanceolata

In this study we tested for trade-offs between the benefit arbuscular mycorrhizal (AM) fungi provide for hosts and their competitive ability in host roots, and whether this potential trade-off shifts in the presence of a plant stress (herbivory). We used three species of AM fungi previously determined to vary in host growth promotion and spore production in association with host plants. We found that these AM fungal species competed for root space, and the best competitor, Scutellospora calospora, was the worst mutualist. In addition, the worst competitor, Glomus white, was the best mutualist. Competition proved to have stronger effects on fungal infection patterns than herbivory, and competitive dominance was not altered by herbivory. We found a similar pattern in a previous test of competition among AM fungi, and we discuss the implications of these results for the persistence of the mutualism and feedbacks between AM fungi and their plant hosts.     

AM真菌对采煤沉陷区黄花菜生长及根际土壤养分的影响

于陕北黄土沟壑采煤沉陷区内布设试验小区,对黄花菜(Hemerocallis citrina Baroni)接种丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)—摩西管柄囊霉菌,通过测定黄花菜光合性能、植株生长、抗逆性、土壤养分含量、根际微生物数量等,揭示AM真菌对黄花菜生长和土壤养分的影响。结果表明,黄花菜种植3—5个月后,接种AM真菌显著提高了黄花菜株高、冠幅及其根系菌根侵染率、菌丝密度。与不接种对照区相比,接种AM真菌后黄花菜叶片的光合速率、可溶性糖含量和过氧化氢酶活性分别提高了51%、12%、79%。接种AM真菌处理区黄花菜根际土壤的电导率、有机质、碱解氮和速效钾含量等均显著高于对照区,细菌数量和磷酸酶活性的菌根贡献率分别达77%和24%。表明采煤沉陷区扰动土壤接种AM真菌具有增强土壤微生物活性、改善土壤肥力和提高黄花菜植株抗逆性的作用,对促进陕北黄土沟壑采煤沉陷区经济作物生长和提高土壤质量具有重要现实生态意义。     

Arbuscular mycorrhizal (AM) technology for the conservation of <Emphasis Type="Italic">Curculigo orchioides</Emphasis> Gaertn.: an endangered medicinal herb

Curculigo orchioides Gaertn. (family Hypoxidaceae) is an endangered anticarcinogenic and aphrodisiac herb, native of India. This study reports the effect of three arbuscular mycorrhizal (AM) fungal inocula on post-transplanting performance of ‘in vitro’ raised C. orchioides plantlets. The three AM fungal inocula consisted of two monospecific cultures of Glomus geosporum and G. microcarpum and one crude consortium of AM fungal spores isolated from rhizosphere soil of C. orchioides growing in natural habitat. Complete plantlets of C. orchioides were raised by direct organogenesis of leaf explants on half strength Murashige and Skoog’s medium devoid of any growth hormone. C. orchioides plantlets responded significantly different to all three mycorrhizal treatments. Mycorrhization enhanced the survival rate of C. orchioides plantlets to 100%. The inoculated plantlets fared significantly better than the uninoculated ones in terms of biomass production and number of leaves and roots per plant. Mycorrhizal plantlets exhibited higher concentrations of photosynthetic pigments as well as minerals P, Mg, Cu, Zn, Mn and Fe in both shoots and roots. Among the three inocula tested, plantlets inoculated with the mixed consortium of AM fungi consistently performed better in terms of the parameters evaluated. The study suggests use of mixed consortium of AM fungi over monospecific cultures for the sustainable cultivation and conservation of endangered medicinal plant: Curculigo orchioides.     

Effect of Tilemsi phosphate rock-solubilizing microorganisms on phosphorus uptake and yield of field-grown wheat (Triticum aestivum L.) in Mali

With the broad aim of biologically improving P uptake by wheat fertilized with Tilemsi phosphate rock (TPR), we investigated the effect of inoculation with TPR-solubilizing microorganisms isolated from Malian soils and with a commercial isolate of the arbuscular mycorrhizal (AM) fungus Glomus intraradices (Gi). AM root length colonization, and growth yield and P concentration of the cultivar Tetra of wheat were measured under field conditions in Mali. Experimental plots were established in Koygour (Diré) during the 2001–2002 cropping season. Inoculation treatments included two fungal isolates, Aspergillus awamori (C1) and Penicillium chrysogenum (C13), and an isolate of Pseudomonas sp. (BR2), used alone or in fungus-bacterium combinations in the presence or absence of the AM fungus Gi. In fertilized treatments, 0 or 30 kg P ha⁻¹ was applied as TPR or diammonium phosphate (DAP). In 45-day-old wheat plants, the highest root length AM colonization (62%) was observed with TPR fertilized wheat inoculated with Gi and BR2. Our results suggest that BR2 is a mycorrhizal-helper bacteria and a good plant growth-promoting rhizobacteria. In fact, inoculation of wheat Tetra fertilized with TPR with a combination of Gi, BR2 and C1 produced the best grain yield with the highest P concentration. This work shows that by inoculating seeds with TPR-solubilizing microorganisms and AM fungi under field conditions in Mali it is possible to obtain wheat grain yields comparable to those produced by using the expensive DAP fertilizer.     

Combined inoculation with Glomus intraradices and Rhizobium tropici CIAT899 increases phosphorus use efficiency for symbiotic nitrogen fixation in common bean (Phaseolus vulgaris L.)

This study compared the response of common bean (Phaseolus vulgaris L.) to arbuscular mycorrhizal fungi (AMF) and rhizobia strain inoculation. Two common bean genotypes i.e. CocoT and Flamingo varying in their effectiveness for nitrogen fixation were inoculated with Glomus intraradices and Rhizobium tropici CIAT899, and grown for 50 days in soil–sand substrate in glasshouse conditions. Inoculation of common bean plants with the AM fungi resulted in a significant increase in nodulation compared to plants without inoculation. The combined inoculation of AM fungi and rhizobia significantly increased various plant growth parameters compared to simple inoculated plants. In addition, the combined inoculation of AM fungi and rhizobia resulted in significantly higher nitrogen and phosphorus accumulation in the shoots of common bean plants and improved phosphorus use efficiency compared with their controls, which were not dually inoculated. It is concluded that inoculation with rhizobia and arbuscular mycorrhizal fungi could improve the efficiency in phosphorus use for symbiotic nitrogen fixation especially under phosphorus deficiency.     

Impact of two fluorescent pseudomonads and an arbuscular mycorrhizal fungus on tomato plant growth,root architecture and P acquisition

The ability of fluorescent pseudomonads and arbuscular mycorrhizal fungi (AMF) to promote plant growth is well documented but knowledge of the impact of pseudomonad-mycorrhiza mixed inocula on root architecture is scanty. In the present work, growth and root architecture of tomato plants (Lycopersicon esculentum Mill. cv. Guadalete), inoculated or not with Pseudomonas fluorescens 92rk and P190r and/or the AMF Glomus mosseae BEG12, were evaluated by measuring shoot and root fresh weight and by analysing morphometric parameters of the root system. The influence of the microorganisms on phosphorus (P) acquisition was assayed as total P accumulated in leaves of plants inoculated or not with the three microorganisms. The two bacterial strains and the AMF, alone or in combination, promoted plant growth. P. fluorescens 92rk and G. mosseae BEG12 when co-inoculated had a synergistic effect on root fresh weight. Moreover, co-inoculation of the three microorganisms synergistically increased plant growth compared with singly inoculated plants. Both the fluorescent pseudomonads and the myco-symbiont, depending on the inoculum combination, strongly affected root architecture. P. fluorescens 92rk increased mycorrhizal colonization, suggesting that this strain is a mycorrhization helper bacterium. Finally, the bacterial strains and the AMF, alone or in combination, improved plant mineral nutrition by increasing leaf P content. These results support the potential use of fluorescent pseudomonads and AMF as mixed inoculants for tomato and suggest that improved tomato growth could be related to the increase in P acquisition.     

Studies on initiation and development of the partner association inGeosiphon pyriforme (Kütz.) v. Wettstein,a unique endocytobiotic system of a fungus (Glomales) and the cyanobacteriumNostoc punctiforme (Kütz.) Hariot

Summary Phagocytosis ofNostoc filaments byGeosiphon, a fungus closely related to AM forming Glomales, was observed under light microscopes. Incorporation can only be performed ifNostoc primordia come into contact with growing hyphal tips of the fungus. The fungal wall just below the apex softens, and fungal cytoplasm is bulged out repeatedly covering the vegetativeNostoc cells but not the heterocytes. New heterocytes are differentiated by the internalised filament whose cells can increase up to ten times in volume after recovering from incorporation strain. TheNostoc cells are coated stepwise by short finger-shaped protuberances of the fungal hypha. These hernia-like outgrowths first remain separated, after 1 to 2 days they merge. Adjacent hyphal walls inside the complex covering disintegrate. Periphal fungal wall portions are united to form a smooth strong outer envelope. Internalisation is categorised as phagocytosis. The partnership is partly specific,Nostoc strains capable of living endocytobiotically are often partners in other symbioses besidesGeosiphon.Abbreviations AM arbuscular mycorrhiza (formerly VAM vesicular arbuscular mycorrhiza) - DIC differential interference contrast - LD light/dark Dedicated to Prof. Dr. Dr. h.c. Eberhard Schnepf on the occasion of his retirement     

Evidence for specificity of cultivable bacteria associated with arbuscular mycorrhizal fungal spores

Bacteria associated with arbuscular mycorrhizal (AM) fungal spores may play functional roles in interactions between AM fungi, plant hosts and defence against plant pathogens. To study AM fungal spore-associated bacteria (AMB) with regard to diversity, source effects (AM fungal species, plant host) and antagonistic properties, we isolated AMB from surface-decontaminated spores of Glomus intraradices and Glomus mosseae extracted from field rhizospheres of Festuca ovina and Leucanthemum vulgare. Analysis of 385 AMB was carried out by fatty acid methyl ester (FAME) profile analysis, and some also identified using 16S rRNA gene sequence analysis. The AMB were tested for capacity to inhibit growth in vitro of Rhizoctonia solani and production of fluorescent siderophores. Half of the AMB isolates could be identified to species (similarity index 0.6) within 16 genera and 36 species. AMB were most abundant in the genera Arthrobacter and Pseudomonas and in a cluster of unidentified isolates related to Stenotrophomonas. The AMB composition was affected by AM fungal species and to some extent by plant species. The occurrence of antagonistic isolates depended on AM fungal species, but not plant host, and originated from G. intraradices spores. AM fungal spores appear to host certain sets of AMB, of which some can contribute to resistance by AM fungi against plant pathogens.     

Presence of different arbuscular mycorrhizal infection patterns in roots of<Emphasis Type="Italic"> Lotus glaber</Emphasis> plants growing in the Salado River basin

Morphological types of arbuscular mycorrhizal (AM) fungi associated with Lotus glaber in sodic soils of the Salado River basin were studied. At least eight colonization patterns (IP) of AM fungi in roots of L. glaber were observed after 30 plants were analyzed. Arum- and Paris-type infection were found in the same plant species. This result supports the idea that AM morphology is not solely under plant control, but is also influenced by fungal identity. One infection pattern, presumably corresponding to Glomus intraradices, and a second, possibly assignable to Glomus tenue, were the most commonly found. Our results reinforce previous suggestions that G. intraradices is well adapted to sodic-saline conditions and may play a role in the resistance of L. glaber to these soils.     

Interactions between arbuscular mycorrhizal fungi and bacteria and their potential for stimulating plant growth

Arbuscular mycorrhizal (AM) fungi and bacteria can interact synergistically to stimulate plant growth through a range of mechanisms that include improved nutrient acquisition and inhibition of fungal plant pathogens. These interactions may be of crucial importance within sustainable, low-input agricultural cropping systems that rely on biological processes rather than agrochemicals to maintain soil fertility and plant health. Although there are many studies concerning interactions between AM fungi and bacteria, the underlying mechanisms behind these associations are in general not very well understood, and their functional properties still require further experimental confirmation. Future mycorrhizal research should therefore strive towards an improved understanding of the functional mechanisms behind such microbial interactions, so that optimized combinations of microorganisms can be applied as effective inoculants within sustainable crop production systems. In this context, the present article seeks to review and discuss the current knowledge concerning interactions between AM fungi and plant growth-promoting rhizobacteria, the physical interactions between AM fungi and bacteria, enhancement of phosphorus and nitrogen bioavailability through such interactions, and finally the associations between AM fungi and their bacterial endosymbionts. Overall, this review summarizes what is known to date within the present field, and attempts to identify promising lines of future research.     

Arbuscular mycorrhizal fungi: a specialised niche for rhizospheric and endocellular bacteria

Arbuscular mycorrhizal (AM) fungi produce an extensive hyphal network which develops in the soil, producing a specialised niche for bacteria. The aim of this paper is to review briefly the interactions shown by these symbiotic fungi with two bacterial groups: (i) the plant-growth promoting rhizobacteria (PGPRs) which are usually associated with fungal surfaces in the rhizosphere, and (ii) a group of endocellular bacteria, previously identified as being related to Burkholderia on the basis of their ribosomal sequence strains. The endobacteria have been found in the cytoplasm of some isolates of AM fungi belonging to Gigasporaceae and offer a rare example of bacteria living in symbiosis with fungi. This revised version was published online in August 2006 with corrections to the Cover Date.     

Liberation of amino acids by heterotrophic nitrogen fixing bacteria

Summary. Large amounts of amino acids are produced by nitrogen-fixing bacteria such as Azotobacter, Azospirillum, Rhizobium, Mesorhizobium and Sinorhizobium when growing in culture media amended with different carbon and nitrogen sources. This kind of bacteria live in close association with plant roots enhanced plant growth mainly as a result of their ability to fix nitrogen, improving shoot and root development suppression of pathogenic bacteria and fungi, and increase of available P concentration. Also, it has been strongly evidenced that production of biologically substances such as amino acids by these rhizobacteria are involved in many of the processes that explain plant-grown promotion. This paper reviews literature concerning amino acids production by nitrogen-fixing bacteria. The role of amino acids in microbial interactions in the rhizosphere and establishment of plant bacterial association is also discussed.     

Arbuscular mycorrhizal fungal species suppress inducible plant responses and alter defensive strategies following herbivory

In a greenhouse experiment using Plantago lanceolata, plants grown with different arbuscular mycorrhizal (AM) fungal species differed in constitutive levels of chemical defense depending on the species of AM fungi with which they were associated. AM fungal inoculation also modified the induced chemical response following herbivory by the specialist lepidopoteran herbivore Junonia coenia, and fungal species varied in how they affected induced responses. On average, inoculation with AM fungi substantially reduced the induced chemical response as compared with sterile controls, and inoculation with a mixture of AM fungi suppressed the induced response of P. lanceolata to herbivory. These results suggest that AM fungi can exert controlling influence over plant defensive phenotypes, and a portion of the substantial variation among experimental tests of induced chemical responses may be attributable to AM fungi.     

Antifungal and Root Surface Colonization Properties of GFP-Tagged Paenibacillus brasilensis PB177

Summary This study evaluates the potential of Paenibacillus brasilensis strain PB177 to inhibit phytopathogenic fungi commonly causing maize diseases and to colonize maize plants. In vitro assays demonstrated antagonistic activity against the fungal pathogens, Fusarium moniliforme and Diplodia macrospora. The PB177 strain was tagged with the gfp gene, encoding the green fluorescent protein (GFP) and GFP-tagged bacteria were detected attached to maize roots by stereo- and confocal microscopy. The GFP-tagged bacteria were also used to treat maize seeds before challenging the seeds with two phytopathogenic fungi. The results demonstrated that the bacterial cells are mobilized to the maize roots in the presence of the fungal pathogens. The ability of P. brasilensis PB177 to inhibit fungal growth in vitro and its capability of colonization of maize roots in vivo suggest a potential application of this strain as a biological control agent. This is the first report on the successful introduction of the GFP marker gene into a P. brasilensis strain, enabling the direct observation of these promising plant growth promoting bacteria on maize roots in situ.     

接种AM菌对西部黄土区采煤沉陷地柠条生长和土壤的修复效应

以采煤沉陷区柠条为宿主植物,研究接种丛枝菌根真菌(arbuscular mycorrhizal fungi,简称AM菌)对柠条生长和根际土壤的改良效应。结果表明:8月份接种AM菌比不接菌柠条的株高、冠幅和地径显著增加了29.11%,29.83%和14.81%,9月份接菌区柠条的根长、平均直径、根表面积和根体积分别比对照区增加了151.0%,34.2%,116.0%和129.3%。接种AM菌增强柠条的抗逆性,接菌区的柠条叶片可溶性糖含量和过氧化氢酶活性分别比对照区增加了13.4%和111.1%。8月份接种AM菌改善了土壤的生物理化性质,接菌区有机质、碱解氮、速效磷和速效钾比对照区分别增加7.06g/kg,140.0 mg/kg,1.82 mg/kg和16.72mg/kg,接种AM菌显著增加了根际土壤中真菌、放线菌、细菌数量和酸性磷酸酶活性。总之,接种AM菌促进采煤沉陷区柠条的生长和土壤的改良。     

Interactions between<Emphasis Type="Italic"> Trichoderma pseudokoningii</Emphasis> strains and the arbuscular mycorrhizal fungi<Emphasis Type="Italic"> Glomus mosseae</Emphasis> and<Emphasis Type="Italic"> Gigaspora rosea</Emphasis>

The interaction between Trichoderma pseudokoningii (Rifai) 511, 2212, 741A, 741B and 453 and the arbuscular mycorrhizal fungi Glomus mosseae (Nicol. & Gerd.) Gerdemann & Trappe BEG12 and Gigaspora rosea Nicolson & Schenck BEG9 were studied in vitro and in greenhouse experiments. All T. pseudokoningii strains inhibited the germination of G. mosseae and Gi. rosea except the strain 453, which did not affect the germination of Gi. rosea. Soluble exudates and volatile substances produced by all T. pseudokoningii strains inhibited the spore germination of G. mosseae. The germination of Gi. rosea spores was inhibited by the soluble exudates produced by T. pseudokoningii 2212 and 511, whereas T. pseudokoningii 714A and 714B inhibited the germination of Gi. rosea spores by the production of volatile substances. The strains of T. pseudokoningii did not affect dry matter and percentage of root length colonization of soybean inoculated with G. mosseae, except T. pseudokoningii 2212, which inhibited both parameters. However, all T. pseudokoningii strains decreased the shoot dry matter and the percentage of AM root length colonization of soybean inoculated with Gi. rosea. The saprotrophic fungi tested seem to affect AM colonization of root by effects on the presymbiotic phase of the AM fungi. No influence of AM fungi on the number of CFUs of T. pseudokoningii was found. The effect of saprotrophic fungi on AM fungal development and function varied with the strain of the saprotrophic species tested.     

丛枝菌根真菌与根围促生细菌相互作用的效应与机制

丛枝菌根(arbuscular mycorrhiza,AM)真菌是植物活体营养专性共生菌,广泛存在于陆地各生态系统中.研究表明,AM真菌与根围促生细菌(plant growth promoting rhizobacteria,PGPR)之间的相互作用,尤其是它们之间的协同作用不仅影响植物养分吸收利用、病原物发生发展、土壤理化特性与生物修复等,而且对于可持续农、林、牧业生产、稳定生态系统都具有十分重要的意义.因此,近年来给予众多关注和研究.综述了AM真菌与PGPR之间的相互影响及其可能的作用机制,以及AM真菌与PGPR协同改善植物营养和生长、协同抑制病原菌、协同修复土壤方面的作用,旨在总结AM真菌与PGPR相互作用的效应与机制方面的最新研究进展,为今后研究发展提供依据.