Interactions Between Plant Growth Promoting Fungi and Arbuscular Mycorrhizal Fungus Glomus Mosseae and Induction of Systemic Resistance to Anthracnose Disease in Cucumber

Cucumber plants were treated with plant growth promoting fungi (PGPF), Phoma sp. (isolates GS8-2 and GS8-3) and Penicillium simplicissimum (isolate GP17-2) with or without the arbuscular mycorrhizal fungus (AMF) Glomus mosseae. Induction of systemic resistance in cucumber against the anthracnose disease caused by Colletotrichum orbiculare was tested to evaluate the nature of the interaction between the PGPF and AMF. Root colonizing ability of each fungal species as influenced by their interaction was also evaluated. Plant roots were pre-inoculated with each PGPF isolate and/or G. mosseae for four weeks and leaves were then challenge inoculated with the pathogen C. orbiculare. Plants treated with each PGPF isolate showed considerable protection against the disease, but the treatment of G. mosseae had no significant effect on disease development. However, combined inoculation of Phoma GS8-2 or GS8-3 with G. mosseae reduced the level of disease protection induced by single inoculation of each Phoma isolate. In contrast, the high levels of protection induced by the P. simplicissimum GP17-2 were not altered by combining it with G. mosseae. Root colonization of both Phoma sp. isolates was also suppressed by the presence of the G. mosseae, but such an effect was not found on the population development of P. simplicissimum. The percent cucumber root length colonized by G. mosseae was not affected by any of the PGPF isolates tested.     

Mass production of Glomus mosseae spores

Five crops inoculated with Glomus mosseae were grown for 10 weeks and the development of mycorrhizal infection and sporulation were assessed. Infected roots from pot cultures of different ages were used to examine the host effect on the development of mycorrhizae. The effectiveness of each host was assessed by measuring spore numbers. For all hosts, the percentage of root length infected increased rapidly up to 10 weeks after sowing. Infectivity of root inocula increased with increasing percentage of root length infected with the inoculum for all crops, except where large numbers of mature spores (1755) had been produced on barley. The highest spore numbers were achieved in the rhizosphere of barley plants, followed by chickpea and beans. The lowest spore numbers were found in the rhizosphere of corn and okra plants. The type of the crop as well as the harvest date greatly influenced the size of the spore population and the extent of root colonization of G. mosseae.     

Absence of VA colonization in Oxalis pes-caprae inoculated with Glomus mosseae

Although members of the Oxalidaceae family have been described as host plants of vesicular-arbuscular mycorrhizal fungi, Oxalis pes-caprae did not become colonized by Glomus mosseae. Extracts of Ox. pes-caprae root inhibited the germination of G. mosseae spores. However, the presence of G. mosseae in the rhizosphere of Ox. pes-caprae produced browning of the roots, which was interpreted as a hypersensitivity response of the plant to the presence of VA fungus.     

Presence of specific polypeptides in onion roots colonized by Glomus mosseae

We studied changes in gene expression during the establishment of vesicular-arbuscular (VA) mycorrhizal symbiosis. Polypeptides were obtained by in vitro translation of total root RNA extracted from VA-colonized and noncolonized root-tissue of onion (Allium cepa L. cv. Babosa), and resolved by two-dimensional polyacrylamide gel electrophoresis. VA mycorrhization led to a specific appearance of eight new polypeptides, and the disappearance of seven polypeptides in VA-colonized root. Our findings indicate that gene expression is altered in response to morphological and physiological changes resulting from the establishment of VA mycorrhizas.     

丛枝菌根真菌促进南美蟛蜞菊生长及对难溶磷的吸收

为了解2种丛枝菌根真菌(AMF)摩西管柄囊霉(Funneliformis mosseae, FM)和地表球囊霉(Glomus versiforme, GV)对入侵植物南美蟛蜞菊(Wedelia trilobata)的生长和对难溶性磷酸盐利用的影响,采用沙培盆栽方式,研究了南美蟛蜞菊在接种AMF与添加难溶性磷酸盐的生长和磷含量的变化。结果表明,在磷限制环境下FM对南美蟛蜞菊的侵染率达55%～69%,GV的侵染率达到63%～80%。添加难溶性磷酸盐后,2种AMF均促进了南美蟛蜞菊茎的伸长(FM:+46%; GV:+65%)、总生物量的增加(FM:+27.2%; GV:+40%)和磷含量的增加(FM:+36.6%; GV:+40.7%)。对比FM,GV对植物利用难溶性磷有更显著的促进作用。因此,南美蟛蜞菊与2种AMF形成的共生体系可以促进植物生长和对营养资源的利用,提高对难溶性磷的吸收效率可能使得南美蟛蜞菊在营养贫乏的环境中更好地建立种群。     

Xyloglucanases in the interaction between saprobe fungi and the arbuscular mycorrhizal fungus Glomus mosseae

We studied the production of xyloglucanase enzymes of pea and lettuce roots in the presence of saprobe and arbuscular mycorrhizal (AM) fungi. The AM fungus Glomus mosseae and the saprobe fungi Fusarium graminearum, Fusarium oxysporum-126, Trichoderma harzianum, Penicillium chrysogenum, Pleurotus ostreatus and Aspergillus niger were used. G. mosseae increased the shoot and root dry weight of pea but not of lettuce. Most of the saprobe fungi increased the level of mycorrhization of pea and lettuce, but only P. chrysogenum and T. harzianum inoculated together with G. mosseae increased the dry weight of pea and lettuce respectively. The AM and saprobe fungi increased the production of xyloglucanases by plant roots. The level of xyloglucanase activities and the number of xyloglucanolytic isozymes in plants inoculated with G. mosseae and most of the saprobe fungi tested were higher than when both microorganisms were inoculated separately. The possible relationship between xylogucanase activities and the ability of AM and saprobe fungi to improve the dry weight and AM root colonization of plants was discussed.     

Interaction of Glomus mosseae and Paecilomyces lilacinus on Meloidogyne javanica of tomato

The effects of Glomus mosseae and Paecilomyces lilacinus on Meloidogyne javanica of tomato were tested in a greenhouse experiment. Chicken layer manure was used as a carrier substrate for the inoculum of P. lilacinus. The following parameters were used: gall index, average number of galls per root system, plant height, shoot and root weights. Inoculation of tomato plants with G. mosseae did not markedly increase the growth of infected plants with M. javanica. Inoculation of plants with G. mosseae and P. lilacinus together or separately resulted in similar shoots and plant heights. The highest root development was achieved when mycorrhizal plants were inoculated with P. lilacinus to control root-knot nematode. Inoculation of tomato plants with G. mosseae suppressed gall index and the average number of galls per root system by 52% and 66%, respectively, compared with seedlings inoculated with M. javanica alone. Biological control with both G. mosseae and P. lilacinus together or separately in the presence of layer manure completely inhibited root infection with M. javanica. Mycorrhizal colonization was not affected by the layer manure treatment or by root inoculation with P. lilacinus. Addition of layer manure had a beneficial effect on plant growth and reduced M. javanica infection.     

Early developmentally regulated genes in the arbuscular mycorrhizal fungus Glomus mosseae: identification of GmGIN1, a novel gene with homology to the C-terminus of metazoan hedgehog proteins

The life cycle of the obligate biotrophic arbuscular mycorrhizal fungi comprises several well-defined developmental stages whose genetic determinants are still unknown. With the aim of understanding the molecular processes governing the early developmental phase of the AM fungal life cycle, a subtractive cDNA library was constructed using a suppressive subtractive hybridization technique. The library contains more than 600 clones with an average size of 500 bp. The isolated cDNAs correspond to genes up-regulated during the early development of the AM fungus Glomus mosseaeversus genes expressed in extraradical hyphae. The expression of several of the isolated genes was further confirmed by RT-PCR analysis. Among the isolated clones, a novel gene named GmGIN1 only expressed during early development in G. mosseae was found. The full-length GmGIN1 cDNA codes for a protein of 429 amino acids. The most interesting feature of the deduced protein is its two-domain structure with a putative self-splicing activity. The N-terminal domain shares sequence similarity with a novel family of GTP binding proteins while the C-terminus has a striking homology to the C-terminal part of the hedgehog protein family from metazoa. The C-terminal part of hedgehog proteins is known to participate in the covalent modification of the N-terminus by cholesterol, and in the self-splicing activity which renders the active form of the protein with signalling function. We speculate that the N-terminal part of GmGIN1, activated through a similar mechanism to the hedgehog proteins, has GTP-binding activity and participates in the signalling events prior to symbiosis formation.     

Alterations in Root Exudation of Intercropped Tomato Mediated by the Arbuscular Mycorrhizal Fungus Glomus mosseae and the Soilborne Pathogen Fusarium oxysporum f.sp. lycopersici

Arbuscular mycorrhizal fungi (AMF) can control soilborne diseases such as Fusarium oxysporum f.sp. lycopersici (Fol). Root exudates play an important role in plant–microbe interactions in the rhizosphere, especially, in the initial phase of these interactions. In this work, we focus on (i) elucidating dynamics in root exudation of Solanum lycopersicum L. in an intercropping system due to AMF and/or Fol; (ii) its effect on Fol development in vitro; and (iii) the testing of the root exudate compounds identified in the chromatographic analyses in terms of effects on fungal growth in in vitro assays. GC‐MS analyses revealed an AMF‐dependent increase in sugars and decrease in organic acids, mainly glucose and malate. In the HPLC analyses, an increase in chlorogenic acid was evident in the combined treatment of AMF and Fol, which is to our knowledge the first report about an increase in chlorogenic acid in root exudates of AM plants challenged with Fol compared with plants inoculated with AMF only, clearly indicating changes in root exudation due to AMF and Fol. Root exudates of AMF tomato plants stimulate the germination rate of Fol, whereas the co‐inoculation of AMF and Fol leads to a reduction in spore germination. In the in vitro assays, citrate and chlorogenic acid could be identified as possible candidates for the reduction in Fol germination rate in the root exudates of the AMF+Fol treatment because they proved inhibition at concentrations naturally occurring in the rhizosphere.     

Dual inoculation with Aspergillus fumigatus and Glomus mosseae enhances biomass production and nutrient uptake in wheat (Triticum aestivum L.) supplied with organic phosphorus as Na-phytate

In a pot experiment, wheat was grown for 50 days in two heat-sterilized low-phosphorus (P) soils supplied with organic P as Na-phytate. Seed inoculation with the phosphatase-producing fungus (PPF) Aspergillus fumigatus or soil inoculation with the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus mosseae increased shoot and root dry weight and root length, phosphatase activity in the rhizosphere and shoot concentrations of P and to a lesser extent of K and Mg. As a rule, the greatest effects on those parameters were most in the combined inoculation treatment (PPF + VAM). Shoot concentrations of Cu and Zn were only enhanced by VAM, not by PPF. At harvest, depletion of organic P in the rhizosphere soil increased in the order of: sterilized soil < PPF < VAM < PPF + VAM which corresponded with the enhanced P concentrations in the plants. The results demonstrate that organic P in form of Na-Phytate is efficiently used by VAM and that use of organic P can be increased by simultaneous inoculation with phosphatase-producing fungi.     

Interaction between arbuscular mycorrhizal fungus Glomus mosseae and plant growth promoting fungus Phoma sp. on their root colonization and growth promotion of cucumber (Cucumis sativus L.)

Interaction between arbuscular mycorrhizal fungus Glomus mosseae and plant growth promoting fungus Phoma sp. was studied for its effect on their root colonization and plant growth of cucumber. Two isolates of Phoma sp. (GS8-2 and GS8-3) were tested with G. mosseae. The percent root length colonized by G. mosseae was not adversely affected by the presence of Phoma isolates. In contrast, the root colonization of both isolates GS8-2 and GS8-3 in 4-week-old plants was significantly reduced (80.7% and 84.3%, respectively) by added G. mosseae. Inoculating plants with each Phoma isolate significantly increased the shoot dry weight. However, dual inoculation of each Phoma isolate with G. mosseae had no significant effect on growth enhancement.     

Effects of the root-lesion nematode Pratylenchus vulnus and the mycorrhizal fungus Glomus mosseae on the growth of EMLA-26 apple rootstock

The interaction between Pratylenchus vulnus and the endomycorrhizal fungus Glomus mosseae on the growth of EMLA 26 apple rootstock was studied under shadehouse conditions in the field during the first 6 months of growth. Fresh top weights, fresh root weights, and shoot lengths of mycorrhizal plants with and without P. vulnus were significantly higher than those of nonmycorrhizal plants. Addition of P to non-mycorrhizal controls had little overall effect. Mycorrhizal treatments with the nematode showed a significantly lower amount of nematodes per gram of root than nonmycorrhizal treatments with P. vulnus. Root colonization by G. mosseae was not affected by the presence of the nematode. No nutrient deficiencies were detected in foliar analyses, although low levels of K, A1, and Fe were detected in nematode treatments. The highest levels of S, Mg, Mn and Zn were detected in P. vulnus inoculated plants. Mycorrhizal plants had the highest levels of N, Na, P, K, and Fe. The importance of early mycorrhizal infection of EMLA 26 apple root-stock in the presence of the nematode is discussed.     

Uptake Kinetics of Arsenic in Upland Rice Cultivar Zhonghan 221 Inoculated with Arbuscular Mycorrhizal Fungi

Arbuscular mycorrhizal fungi (AMF) appear to be highly associated with arsenic (As) uptake in host plants because arsenate (As(V)) and phosphorus (P) share the same transporter, whereby AMF can enhance P uptake. A short-term experiment was conducted for low- (0 to 0.05 mM As) and high-affinity (0 to 2.5 mM As) uptake systems, to investigate the AMF role on As uptake mechanism in plants, which may explain As uptake kinetics in upland rice cultivar: Zhonghan 221. When concentration of As ranged from 0 to 0.05 mM, Funneliformis geosporum (Fg) significantly decreased arsenite (As(III)) and monomethylarsonicacid (MMA) uptake when (p < 0.05) compared to non-mycorrhizal (NM) treatment, since the major route for (As(III)) in rice roots—rice silicon transporter Lsi1 would be influenced by Fg inoculation at high As concentrations. Fg can also reduce As(V) uptake significantly (p < 0.05) under both uptake systems relative to NM treatment, whereas, Funneliformis mosseae (Fm) increased As(V) and MMA uptake in rice roots, with MMA uptake rate generally lower than As(III) and As(V). Using suitable AMF species inoculation with rice, As uptake and accumulation in rice grains can be reduced and the risk to human health, once consumed, can be minimized.     

Amelioration of Pb and Mn Toxicity to Arbuscular Mycorrhizal Fungus Glomus intraradices by Maize Root Exudates

The effect of maize root exudates on the toxicity of lead and manganese to arbuscular mycorrhizal fungus (AMF) Glomus intraradices was studied in vitro by observation of intraradical hyphae regrowth from colonised root segments. Higher heavy metal (HM) concentrations strongly reduced the hyphal growth, however, the inhibitory effect was to a large extent eliminated by the addition of maize root exudates to the media. However, the capacity of exudates to ameliorate HM toxicity was limited and did not operate when a threshold HM concentration was reached. This revised version was published online in July 2006 with corrections to the Cover Date.     

AM真菌对百合调节激素平衡与细胞渗透性以及改善耐盐性的研究

该试验以东方百合(Lilium brownii)品种‘西伯利亚’和丛枝菌根(AM)真菌摩西斗管囊霉(Funneliformis mosseae)、变形球囊霉(Glomus versiforme)为材料,在温室盆栽条件下,设置NaCl胁迫(0、0.4%、0.8%和1.2%NaCl溶液处理)和接种AM真菌[接种摩西斗管囊霉、变形球囊霉、摩西斗管囊霉+变形球囊霉及未接种对照]双因素试验,分析各处理百合的激素平衡与细胞渗透性变化特征,以明确AM真菌提高百合耐盐性的效应,初步探索AM真菌增强百合耐盐性的作用机制。结果表明:(1)接种AM真菌能有效增加盐胁迫下百合植株的株高和生物量,显著提高百合耐盐系数,双接种处理的株高及地上、地下部分干重在1.2%NaCl胁迫下分别比未接种对照显著增加8.9%、14.5%和11.2%。(2)接种AM真菌能显著提高盐胁迫下百合植株叶片的矿质元素P、K、S含量,显著降低叶片内Na和Fe含量,双接种处理的P、K、S含量在1.2%NaCl胁迫下分别比对照提高10.9%、8.3%、13.7%,而其Na和Fe含量分别显著下降28.4%和66.4%。(3)接种AM真菌能显著提高盐胁迫下百合内源激素吲哚乙酸(IAA)和脱落酸(ABA)含量,双接种处理在1.2%NaCl胁迫下百合内源IAA、ABA含量分别是对照的1.2和1.5倍。(4)接种AM真菌能显著提高盐胁迫下百合可溶性蛋白含量,显著降低其脯氨酸含量,双接种处理在1.2%NaCl胁迫下比对照的升降幅度分别为69%和31%。(5)接种AM真菌能显著降低盐胁迫下百合叶片丙二醛和相对电导率,双接种处理在1.2%NaCl胁迫下比对照分别显著下降58.1%和9.0%。研究发现,AM真菌可以通过增强百合对养分的吸收、降低氧化胁迫造成的伤害、调节植物内源激素平衡状况与细胞渗透性来增强自身的耐盐能力,且双接种处理的效果优于单一接种。     

Influence of Arbuscular Mycorrhizal (AM) Symbiosis on Phosphorus Leaching through Soil Cores

Two experiments with soil cores were carried out to investigate the effects of arbuscular mycorrhizal (AM) fungal colonization on mobility of phosphorus (P) during leaching of repacked columns of a soil with a loamy sand texture. Trifolium subterraneum plants inoculated with an AM fungus or not inoculated were grown in cores with low or high P concentrations for 8 or 10 weeks in the glasshouse. Cores were then irrigated with 2500 mL water and the leachate collected. Plant growth and the amounts of P removed by plants, remaining in soil as available P and removed dissolved in leachate were measured. Mycorrhizal fungal colonization and development of external hyphae were also determined. Inoculation and/or P application significantly increased plant growth and plant P removal and decreased P leaching. In low P soils AM fungal colonization significantly increased plant P uptake and decreased soil available P and total dissolved P in leachates. Lower P leaching from cores with AM plants under low P conditions was related to enhancement of plant growth and to scavenging and removal of P from the soil by roots and/or external hyphae. When P was applied AM effects were not observed and available P remaining in the soil after leaching was much higher, regardless of AM fungal colonization.     

Effects of the root-lesion nematode Pratylenchus vulnus and the vesicular-arbuscular mycorrhizal fungus Glomus mosseae on the growth of three plum rootstocks

The effects of Pratylenchus vulnus and the endomycorrhizal fungus Glomus mosseae on growth of Myrobalan 605, Marianna 2624 and San Julian 655-2 plum rootstocks were measured under shadehouse conditions in the field for two growing seasons (1990–91). Shoot dry weights were higher in the majority of the vesicular-arbuscular mycorrhizal (VAM) alone inoculated plants after both growing seasons. Root weights of mycorrhizal Myrobalan and Marianna were higher than root weights of the same rootstocks lacking mycorrhizae, inoculated with P. vulnus, and VAM in combination with the nematode. Mycorrhizal Marianna inoculated with the nematode showed a considerably higher final nematode population in relation to non-inoculated VAM treatments. No correlation was found in the number of nematodes per gram of root between mycorrhizal and non-mycorrhizal treatments. P. vulnus adversely affected the mycorrhizal colonization in Marianna, but not in Myrobalan and San Julian. Marianna appears to be more mycorrhizal dependent than the two other rootstocks.     

Litter N retention over winter for a low and a high phenolic species in the alpine tundra

Mycorrhizal fungus colonization of roots may modify plant metal acquisition and tolerance. In the present study, the contribution of the extraradical mycelium of an arbuscular mycorrhizal (AM) fungus, Glomus mosseae (BEG 107), to the uptake of metal cations (Cu, Zn, Cd and Ni) by cucumber (Cucumis sativus) plants was determined. The influence of the amount of P supplied to the hyphae on the acquisition and partitioning of metal cations in the mycorrhizal plants was also investigated. Pots with three compartments were used to separate root and root-free hyphal growing zones. The shoot concentration of Cd and Ni was decreased in mycorrhizal plants compared to non-mycorrhizal plants. In contrast, shoot Zn and Cu concentrations were increased in mycorrhizal plants. High P supply to hyphae resulted in decreased root Cu concentrations and shoot Cd and Ni concentrations in mycorrhizal plants. These results confirm that some elements required for plant growth (P, Zn, Cu) are taken up by mycorrhizal hyphae and are then transported to the plants. Conversely, Cd and Ni were transported in much smaller amounts by hyphae to the plant, so that arbuscular mycorrhizal fungus colonization could partly protect plants from toxic effects of these elements. Selective uptake and transport of plant essential elements over non-essential elements by AM hyphae, increased growth of mycorrhizal plants, and metal accumulation in the root may all contribute to the successful growth of mycorrhizal plants on metal-rich substrates. These effects are stimulated when hyphae can access sufficient P in soil.