Interactions of Paecilomyces lilacinus strain 251 with the mycorrhizal fungus Glomus intraradices: Implications for Meloidogyne incognita control on tomato

The interactions of Paecilomyces lilacinus strain 251 with the arbuscular mycorrhizal fungus Glomus intraradices and their significance for the control of Meloidogyne incognita on tomato were investigated in greenhouse experiments. Application of P. lilacinus had no effect on the frequency and intensity of tomato root colonization by G. intraradices. Likewise, the decline of the nematophagous fungus densities after single application in soil was not affected by the presence of the mycorrhizal fungus. Single application of P. lilacinus, as pre-planting soil treatment, resulted in significant reduction of nematode damage. In contrast, mycorrhizal inoculation did not provide sufficient biocontrol. Combined application of the two agents did not enhance root protection compared to single treatments. Double treatment of mycorrhized seedlings with P. lilacinus, as seedling drench and pre-planting soil treatment, 4 and 1 week before transplanting, respectively, resulted in the highest reduction of the nematode damage. These results indicate the potential of the commercial P. lilacinus strain 251 and mycorrhiza for integration in nematode control strategies.     

Accumulation of apocarotenoids in mycorrhizal roots of leek (Allium porrum)

Colonization of the roots of leek (Allium porrum L.) by the arbuscular mycorrhizal fungus Glomus intraradices induced the formation of apocarotenoids, whose accumulation has been studied over a period of 25 weeks. Whereas the increase in the levels of the dominating cyclohexenone derivatives resembles the enhancement of root length colonization, the content of mycorradicin derivatives remains relatively low throughout. Structural analysis of the cyclohexenone derivatives by mass spectrometry and NMR spectroscopy showed that they are mono- and diglycosides of 13-hydroxyblumenol C and blumenol C acylated with 3-hydroxy-3-methyl-glutaric and/or malonic acid. Along with the isolation of three known compounds five others are shown to be hitherto unknown members of the fast-growing family of mycorrhiza-induced cyclohexenone conjugates.     

Arbuscular mycorrhizal colonization of tomato by Gigaspora and Glomus species in the presence of root flavonoids

The effect of flavonoids isolated from arbuscular mycorrhizal (AM) colonized and noncolonized clover roots on the number of entry points and percentage of root colonization of tomato (Lycopersicum esculentum L.) by Gigaspora rosea, Gi margarita, Glomus mosseae and G. intrarradices symbionts was determined. With fungi of both genera, a correlation between the number of entry points and the percentage of root colonization was found in the presence of some of the tested flavonoids. The flavonoids acacetin and rhamnetin, present in AM clover roots, inhibited the formation of AM penetration structures and the AM colonization of tomato roots, whereas the flavonoid 5,6,7,8,9-hydroxy chalcone, which could not be detected in AM clover root, inhibited both parameters. The flavonoid quercetin, which was present in AM clover roots, stimulated the penetration and root colonization of tomato by Gigaspora. However, the flavonoids 5,6,7,8-hydroxy-4'-methoxy flavone and 3,5,6,7,4'-hydroxy flavone, which was not found in AM clover root, increased the number of entry points and the AM colonization of tomato roots by Gigaspora. These results indicated that flavonoids could be imnplicated in the process of regulation of AM colonization in plant root, but its role is highly complex and depend not only on flavonoids, but also on AM fungal genus or even species.     

AM真菌控制蚕豆枯萎病发生的根际微生物效应

在温室盆栽条件下,研究了接种尖孢镰刀菌蚕豆专化型和丛枝菌根(AM)真菌(摩西球囊霉Gm、扭形球囊霉Gt、根内球囊霉Gi及幼套球囊霉Ge)对灭菌连作土壤中蚕豆幼苗生长、枯萎病发生、根际镰刀菌数量和微生物代谢功能多样性的影响.结果表明: 接种AM真菌能显著增加蚕豆幼苗地上部和地下部鲜质量;接种Gm、Gt、Gi和Ge真菌使蚕豆枯萎病病情指数分别显著降低94.0%、60.0%、64.0%和94.0%,使蚕豆根际镰刀菌数量分别显著降低98.6%、74.3%、77.8%和90.4%,以Gm和Ge真菌对蚕豆枯萎病的抑制效应最好.接种Gm、Gt和Ge显著提高了根际微生物对糖类(CH)、氨基酸类(AA)、羧酸类(CA)和酚酸类(PA)碳源的利用,使蚕豆根际微生物的AWCD值分别显著提高34.4%、31.5%和50.8%;而接种Gi对AA、CA、PA类碳源利用和AWCD值均无显著影响,使微生物对CH类碳源的利用显著降低59.3%.主成分和相关分析表明, 接种AM真菌明显改变了蚕豆根际微生物的群落结构.接种Gt真菌提高了蚕豆根际微生物对AA类碳源(β-甲基-D-葡萄糖苷、D-半乳糖酸-γ-内酯、D-甘露醇、N-乙酰基-D-葡萄糖胺、D-纤维二糖)和CA类碳源(D-半乳糖醛酸)的利用,而接种Gi真菌降低了微生物对以上碳源的利用;接种Gm和Ge真菌提高了根际微生物对L-精氨酸和4-羟基苯甲酸的利用.表明糖类和羧酸类是接种Gt真菌后蚕豆根际土壤微生物利用的主要碳源, 氨基酸和酚酸类是接种Gm和Ge后蚕豆根际土壤微生物利用的主要碳源.接种AM真菌显著提高了蚕豆根际微生物的活性,改变了微生物群落结构并抑制病原菌的增殖,是AM真菌减轻了蚕豆枯萎病的发生的重要原因.不同AM真菌减轻了蚕豆枯萎病的发生与其改变微生物的碳源利用密切相关.     

Synergistic effects of antagonistic fungi and a plant growth promoting rhizobacterium, an arbuscular mycorrhizal fungus, or composted cow manure on populations of Meloidogyne incognita and growth of tomato

Effects of four antagonistic fungi (Paecilomyces lilacinus, Pochonia chlamydosporia, Trichoderma harzianum and Gliocladium virens) alone and together with a plant growth promoting rhizobacterium Pseudomonas putida, an arbuscular mycorrhizal fungus Glomus intraradices or with composted cow manure (CCM) were assessed on the growth of tomato and on the reproduction of Meloidogyne incognita in glasshouse experiments. Application of all antagonistic fungi (except G. virens), P. putida, G. intraradices or CCM caused a significant increase in the growth of plants without nematodes. However, use of either of these fungi, P. putida, G. intraradices and CCM against plants with nematodes caused a significant increase in tomato growth. Paecilomyces lilacinus caused a 42% increase in the growth of nematode-inoculated plants followed by P. chlamydosporia (36%), T. harzianum (18%) and G. virens (15%). CCM caused about 57% increase in the growth of nematode-inoculated plants followed by P. putida (37%) and G. intraradices (31%). Maximum increase (71%) in the growth of nematode-inoculated plants was observed when CCM was used with P. lilacinus. Moreover, P. lilacinus caused a high reduction (55%) in galling and nematode multiplication, while G. virens the least (25%). Use of P. putida also caused a 39% reduction in galling and nematode multiplication followed by CCM (34%) and G. intraradices (32%). Combined use of CCM with P. lilacinus caused maximum reduction (79%) in galling and nematode multiplication. Re-isolation of antagonistic fungi from nematodes revealed that P. lilacinus parasitised more females and eggs than other antagonistic fungi. Root colonisation by P. putida was increased with P. lilacinus, while colonisation by G. intraradices was reduced in the presence of antagonistic fungi.     

Measurement of the viability of arbuscular-mycorrhizal fungi using three different stains; relation to growth and metabolic activities of soybean plants

Histochemical staining of alkaline phosphatase (ALP) and succinate dehydrogenase (SDH) activities in four arbuscular mycorrhizal fungi (Glomus intraradices, G. fasciculatum, G. monosporum and G. mosseae) and their relation to growth and metabolic activities of soybean plants were investigated in a greenhouse experiment. In general, mycorrhizal inoculation significantly increased the growth responses, phosphorus and nitrogen contents, acid and alkaline phosphatases as well as total soluble protein of soybean compared to non-mycorrhizal plants. Stimulation was related to the viability of each mycorrhizal fungus. The localization of succinate dehydrogenase (as a vital stain of metabolically active fungus) and alkaline phosphatase activity (as a potential marker of efficiency of the symbiosis) in the arbuscular mycorrhizal fungi were variable. The activity appeared in young arbuscles and intercellular hyphae, whereas the collapsed arbuscules were inactive. The histochemical staining results demonstrated that the activity of alkaline phosphatase fungi was lower than succinate dehydrogenase. The use of nitroblue tetrazolium chloride as a vital stain for SDH activity showed that all mycorrhizal infection revealed by trypan blue staining was not physiologically active. Thus, the possible utilization of these enzymes to assess the activity of mycorrhizal fungi and its relation with effectively for plant growth and mineral contents is discussed.     

Effects of two sterol biosynthesis inhibitor fungicides (fenpropimorph and fenhexamid) on the development of an arbuscular mycorrhizal fungus

The effects of different concentrations (0.2, 2, 20, 200 mg l⁻¹) of two sterol biosynthesis inhibitor (SBI) fungicides, i.e. fenpropimorph and fenhexamid, were evaluated on the spore germination, germ tube elongation, sporulation, and root colonization of Glomus intraradices grown monoxenically in association with transformed carrot roots. The percentage of germinated spores incubated on the SBI fungicides and the length of the germ tubes decreased with increasing concentrations of both fungicides. However, for spore germination this impact was fungistatic rather than fungicidal. Extraradical mycelium architecture and spore production in contact with the SBI fungicides were also strongly impacted at high concentration (20 mg l⁻¹). Conversely, the colonization of roots developing in the fungicide-free compartment, but interconnected with the extraradical mycelium developing on the SBI fungicides, appeared unaffected. Our results demonstrated that the monoxenic culture system could be used as a standardized, reproducible technique to compare the impacts of different molecules on arbuscular mycorrhizal fungi, and for the initial screening of new candidate molecules before registration.     

丛枝菌根真菌对薰衣草耐热性的影响

于不同温度（25℃/20℃、35℃/30℃和40℃/35℃）下测定接种丛枝菌根真菌（arbuscular mycorrhizal fungi,AMF）摩西斗管囊霉Funneliformis mosseae、变形球囊霉Glomus versiforme和F. mosseae+G. versiforme 混合菌种处理对狭叶薰衣草Lavandula angustifolia耐热性的影响。结果表明,供试AMF能与狭叶薰衣草根系形成菌根共生体,以混合菌种处理的侵染率最高,达到68%。40℃/35℃下,与不接种AMF对照相比,混合菌种处理的狭叶薰衣草叶片可溶性糖、可溶性蛋白、脯氨酸和叶绿素等含量以及根系活力分别提高了46%、68%、65%、29%和70%;与不接种AMF对照相比,3种温度处理下接种AMF显著增加了狭叶薰衣草植株超氧化物歧化酶、过氧化物酶、过氧化氢酶、抗坏血酸过氧化物酶和硝酸还原酶活性,而降低相对电导率及丙二醛含量。表明接种AMF能增强狭叶薰衣草抗氧化酶活性,减轻高温造成的伤害,增强耐热性,与单一接种相比以混合接种摩西斗管囊霉和变形球囊霉提高狭叶薰衣草耐热性的效应最大。     

感染羽茅的香柱菌属内生真菌对丛枝菌根真菌孢子萌发的影响

内生真菌与丛枝菌根(AM)真菌是构成草原生态系统的重要组成部分.内生真菌会抑制其宿主植物的AM真菌侵染率.本研究以感染2种香柱菌属内生真菌[Epichloё gansuensis(Eg)和E. sibirica(Es)]的天然禾草羽茅为供试材料,进行体外纯培养的内生真菌培养滤液、感染内生真菌的羽茅叶片(包括鲜叶和枯叶)浸提液,以及根系分泌物对摩西球囊霉(Gm)和幼套球囊霉(Ge)2种AM真菌孢子萌发影响试验.结果表明: 香柱菌属内生真菌的培养滤液会显著抑制2种AM真菌孢子的萌发,而感染香柱菌属内生真菌的羽茅根系分泌物只对Ge孢子萌发有显著抑制作用,且上述抑制作用与内生真菌种类无关;鲜叶浸提液对Gm和Ge的孢子萌发率均无显著影响,而枯叶浸提液对Ge的孢子萌发有显著抑制作用.在自然生态系统中,香柱菌属内生真菌通常存在于宿主植物体内,可能通过影响宿主植物的根系分泌物来影响AM真菌孢子的萌发.     

Calcareous impact on arbuscular mycorrhizal fungus development and on lipid peroxidation in monoxenic roots

The present work underlined the negative effects of increasing CaCO₃ concentrations (5, 10 and 20 mM) both on the chicory root growth and the arbuscular mycorrhizal fungus (AMF) Glomus irregulare development in monoxenic system. CaCO₃ was found to reduce drastically the main stages of G. irregulare life cycle (spore germination, germinative hyphae elongation, root colonization, extraradical hyphae development and sporulation) but not to inhibit it completely. The root colonization drop was confirmed by the decrease in the arbuscular mycorrhizal fungal marker C16:1ω5 amounts in the mycorrhizal chicory roots grown in the presence of CaCO₃. Oxidative damage evaluated by lipid peroxidation increase measured by (i) malondialdehyde (MDA) production and (ii) the antioxidant enzyme peroxidase (POD) activities, was highlighted in chicory roots grown in the presence of CaCO₃. However, MDA formation was significantly higher in non-mycorrhizal roots as compared to mycorrhizal ones. This study pointed out the ability of arbuscular mycorrhizal symbiosis to enhance plant tolerance to high levels of CaCO₃ by preventing lipid peroxidation and so less cell membrane damage.     

亚热带典型树种根毛特征及其与共生真菌的关系

根毛和共生真菌增加了吸收面积,提高了植物获取磷等土壤资源的能力。由于野外原位观测根表微观结构较为困难,吸收细根、根毛、共生真菌如何相互作用并适应土壤资源供应,缺乏相应的数据和理论。该研究以受磷限制的亚热带森林为对象,选取了21种典型树种,定量了根毛存在情况、属性变异,分析了根毛形态特征与共生真菌侵染率、吸收细根功能属性之间的关系,探讨了根表结构对低磷土壤的响应和适应格局。结果表明:1)在亚热带森林根毛不是普遍存在的, 21个树种中仅发现7个树种存有根毛, 4个为丛枝菌根(AM)树种, 3个为外生菌根(ECM)树种。其中,马尾松(Pinus massoniana)根毛出现率最高,为86%;2)菌根类型是理解根-根毛-共生真菌关系的关键,AM树种根毛密度与共生真菌侵染率正相关,但ECM树种根毛直径与共生真菌侵染率负相关; 3) AM树种根毛长度和根毛直径、ECM树种根毛出现率与土壤有效磷含量呈负相关关系。该研究揭示了不同菌根类型树种根毛-共生真菌-根属性的格局及相互作用,为精细理解养分获取策略奠定了基础。     

Differential effects of tropical arbuscular mycorrhizal fungal inocula on root colonization and tree seedling growth: implications for tropical forest diversity

The potential for mycorrhizae to influence the diversity and structuring of plant communities depends on whether their affinities and effects differ across a suite of potential host species. In order to assess this potential for a tropical forest community in Panama, we conducted three reciprocal inoculation experiments using seedlings from six native tree species. Seeds were germinated in sterile soil and then exposed to arbuscular mycorrhizal fungi in current association with naturally infected roots from adults of either the same or different species growing in intact forest. The tree species represent a range of life histories, including early successional pioneers, a persistent understory species, and emergent species, typical of mature forest. Collectively, these experiments show: (i) the seedlings of small-seeded pioneer species were more dependent on mycorrhizal inocula for initial survival and growth; (ii) although mycorrhizal fungi from all inocula were able to colonize the roots of all host species, the inoculum potential (the infectivity of an inoculum of a given concentration) and root colonization varied depending on the identity of the host seedling and the source of the inoculum; and (iii) different mycorrhizal fungal inocula also produced differences in growth depending on the host species. These differences indicate that host–mycorrhizal fungal interactions in tropical forests are characterized by greater complexity than has previously been demonstrated, and suggest that tropical mycorrhizal fungal communities have the potential to differentially influence seedling recruitment among host species and thereby affect community composition.     

丛枝菌根真菌对枳根净离子流及锌污染下枳苗矿质营养的影响

盆栽实验研究了不同施Zn水平(0、300 mg/kg和600 mg/kg)下,接种丛枝菌根真菌Glomus intraradices对枳苗生长、Zn、Cu、P、K、Ca、Mg分布的影响,并采用非损伤微测技术测定分析了菌根化与非菌根化枳根净Ca²⁺、H⁺、NO₃^-离子流动态。结果表明:(1)在不同施Zn水平下,接种菌根真菌显著提高了枳苗地上部及根部鲜重;随着施Zn水平的提高,菌根侵染率呈降低趋势,枳苗地上部与根部Zn含量呈增加趋势,且接种株根部Zn含量显著高于未接种株。(2)接种株未施Zn处理的地上部Cu、P、K、Mg和根部Cu含量、施600 mg/kg Zn处理的根部Cu及施300 mg/kg Zn处理的根部P含量均显著高于对照,而菌根真菌侵染对枳苗Ca含量并无显著性影响。(3)接种株未施Zn处理的根部距根尖端0 μm和600 μm处净Ca²⁺流出速率、600 μm处净H⁺流入速率、2400 μm处净NO₃^-流入速率均显著高于未接种株。     

Arbuscular mycorrhizal colonization of giant sequoia (Sequoiadendron giganteum) in response to restoration practices

Interactions with soil microbiota determine the success of restoring plants to their native habitats. The goal of our study was to understand the effects of restoration practices on interactions of giant sequoia Sequoiadendron giganteum with arbuscular mycorrhizal (AM) fungi (Glomeromycota). Natural regeneration of Sequoiadendron is threatened by the absence of severe fires that create forest canopy gaps. Generating artificial canopy gaps offers an alternative tool for giant sequoia restoration. We investigated the effect of regeneration practices, including (i) sapling location within gaps, (ii) gap size and (iii) soil substrate, on AM fungal colonization of giant sequoia sapling roots in a native giant sequoia grove of the Sierra Nevada, California. We found that the extent of AM fungal root colonization was positively correlated with sapling height and light availability, which were related to the location of the sapling within the gap and the gap size. While colonization frequency by arbuscules in saplings on ash substrate was higher relative to saplings in mineral soil, the total AM fungal root colonization was similar between the substrates. A negative correlation between root colonization by Glomeromycota and non-AM fungal species indicated antagonistic interactions between different classes of root-associated fungi. Using DNA genotyping, we identified six AM fungal taxa representing genera Glomus and Ambispora present in Sequoiadendron roots. Overall, we found that AM fungal colonization of giant sequoia roots was associated with availability of plant-assimilated carbon to the fungus rather than with the AM fungal supply of mineral nutrients to the roots. We conclude that restoration practices affecting light availability and carbon assimilation alter feedbacks between sapling growth and activity of AM fungi in the roots.     

A mycorrhizal fungus changes microtubule orientation in tobacco root cells

Summary Cortical cells of mycorrhizal roots undergo drastic morphological changes, such as vacuole fragmentation, nucleus migration, and deposition of cell wall components at the plant-fungus interface. We hypothesized that the cytoskeleton is involved in these mechanisms leading to cell reorganization. We subjected longitudinal, meristem to basal zone, sections of uninfectedNicotiana tabacum roots to immunofluorescence methods to identify the microtubular (MT) structures associated with root cells. Similar sections were obtained from tobacco roots grown in the presence ofGigaspora margarita, an arbuscular mycorrhizal fungus which penetrates the root via the epidermal cells, but mostly develops in the inner cortical cells. While the usual MT structures were found in uninfected roots (e.g., MTs involved in mitosis in the meristem and cortical hoops in differentiated parenchyma cells), an increase in complexity of MT structures was observed in infected tissues. At least three new systems were identified: (i) MTs running along large intracellular hyphae, (ii) MTs linking hyphae, (iii) MTs binding the hyphae to the host nucleus. The experiments show that mycorrhizal infection causes reorganization of root MTs, suggesting their involvement in the drastic morphological changes shown by the cortical cells.     

Mycotrophy of Annona cherimola and the morphology of its mycorrhizae

The mycotrophic character of Annona cherimola (Magnoliales), a tropical/subtropical plantation crop of interest, is described for the first time. This crop seems to depend on mycorrhizae (arbuscular) for optimal growth, with Glomus deserticola being the most effective endophyte tested. Study of the morphology of the arbuscular mycorrhizae in Annona roots showed exclusively intracellular hyphal development, with cell-to-cell fungal passage and an abundance of arbuscules and coiled hyphae within cells. Intercellular distributive hyphae were not observed. The morphology and the pattern of spread of the mycorrhizal colonization were similar for the different endophytes involved and appeared to be dependent on the host root. Such features of mycorrhizal colonization are characteristic of host species lacking intercellular air channels and have been described for some species of ecological interest, but they are not commonly noted in the mycorrhizal literature, especially that dealing with crop species. Some ecophysiological consequences of this pattern of colonization are discussed.     

Arbuscular mycorrhiza alter the concentration of essential oils in oregano (Origanum sp., Lamiaceae)

The effect of root colonization by Glomus mosseae on the qualitative and quantitative pattern of essential oils (EO) was determined in three oregano genotypes (Origanum sp.). To exclude a simple P-mediated effect through mycorrhization the effect of P application to plants on the EO accumulation was also tested. In two genotypes the leaf biomass was increased through mycorrhization. Root colonization by the arbuscular mycorrhizal fungus (AMF) did not have any significant effect on the EO composition in oregano; however, in two genotypes the EO concentration significantly increased. As EO levels in P-treated plants were not enhanced, we conclude that the EO increase observed in mycorrhizal oregano plants is not due to an improved P status in mycorrhizal plants, but depends directly on the AMF–oregano plant association.     

Effects of arbuscular mycorrhizal fungi on calorific value and contents of carbon and ash in Robinia pseudoacacia

该试验以根内球囊霉(Glomus intraradices)和地表球囊霉(G. versiforme)为接种剂, 研究了丛枝菌根真菌对刺槐(Robinia pseudoacacia)生物量、热值、含碳量、灰分、能量积累和碳素积累的影响。结果表明, 接种根内球囊霉和地表球囊霉对提高刺槐生物量、热值、能量积累和碳素积累都起到了重要作用。接种根内球囊霉和地表球囊霉后刺槐的总生物量比对照分别增加了89.61%和91.34%, 能量积累分别比对照增加102.20%和94.19%, 碳素积累分别比对照增加93.30%和77.21%; 同时发现刺槐的能量和碳主要分布在根系和叶, 而茎中能量和碳所占的比例较小。接种根内球囊霉提高了刺槐的干重热值, 其根、茎、叶的干重热值分别比对照增加7.72%、8.94%和8.41%; 接种地表球囊霉也显著(p < 0.05)提高了刺槐的干重热值, 但其效果低于根内球囊霉。接种根内球囊霉显著(p < 0.05)提高了刺槐根的含碳量, 对茎和叶的含碳量影响不明显。接种根内球囊霉和地表球囊霉都显著(p < 0.05)提高了刺槐茎和叶的去灰分热值。