The interface between the arbuscular mycorrhizal fungus Glomus intraradices and root cells of Panax quinquefolius: a Paris-type mycorrhizal association

Two major types of arbucular mycorrhizal associations, the Arum-type and the Paris-type, have been identified based on morphological features. Although the Paris-type is the most common, it is the Arum-type that has been most intensively studied in terms of structure/function because of its prevalence in agronomically important plant species. In this study, the interface between the host cell cytoplasm and intracellular hyphae (extensive hyphal coils and arbusculate coils), which typify the Paris-type mycorrhiza, was studied. Using immunofluorescence techniques combined with laser scanning confocal microscopy, dramatic changes in the cytoskeleton in colonized cells were observed. Changes in the positioning of both host cell microtubules and actin filaments occurred in colonized plant cells. Both microtubules and actin filaments were associated with the hyphal coils and the arbusculate coils. An interfacial matrix, of host origin, was demonstrated between hyphal coils and arbusculate coils using various affinity techniques. It formed an apoplastic compartment consisting of cellulose and pectins between the fungus and host cell cytoplasm. There was less labelling adjacent to the fine branches of arbusculate coils compared to the hyphal coils. These observations show some similarities to those seen with Arum-type mycorrhizas.     

Morphological types of arbuscular mycorrhizal fungi in roots of understory plants in Japanese deciduous broadleaved forests

Morphological types of arbuscular mycorrhizal (AM) fungi in roots of understorey plants were examined in three different Japanese deciduous broadleaved forests. In total, 43 species belonging to 33 genera from 27 families were examined for the morphological types of AM. The number of flowering plant species having Paris-type AM was greater than those having Arum-type AM in each plot. This tendency was more prominent in herbaceous plants than woody plants with nine species having Paris-type associations among ten herbaceous plant species examined. Therefore, it is suggested from the ecological point of view that Paris-type associations could be advantageous for the herbaceous understorey plants growing slowly in these environments. The influence of plant identity on the morphological types of AM was also discussed by arranging the plants examined with the morphological types in a current plant phylogeny scheme. In this study, some new records on the morphological types of AM in some new plant families were obtained including the first report of a typical Arum-type AM in gymnosperms.     

Positive association between mycorrhiza and foliar endophytes in Poa bonariensis, a native grass

The interaction between mycorrhiza and leaf endophytes (Neotyphodium sp.) was studied in three Poa bonariensis populations, a native grass, differing significantly in endophyte infection. The association between endophytes and mycorrhizal fungi colonisation was assessed by analysing plant roots collected from the field. We found that roots from endophyte-infected populations showed a significantly higher frequency of colonisation by mycorrhizal fungi and that soil parameters were not related to endophyte infection or mycorrhiza colonization. In addition, we did not observe significant differences in the number of AM propagules in soils of the three populations sites. We also report the simultaneous development of Paris-type and Arum-type mycorrhiza morphology within the same root systems of P. bonariensis. The co-occurrence of both colonisation types in one and the same root system found in the three populations, which differed in Neotyphodium infection, suggests that foliar endophytes do not determine AM morphology. The percentage of root length colonised by different types of fungal structures (coils, arbuscules, longitudinal hyphae and vesicles) showed significant and positive differences in arbuscular frequency associated with endophyte infection, whereas the much smaller amounts of vesicles and hyphal coils did not differ significantly.     

大兴安岭落叶松林丛枝菌根真菌多样性

对大兴安岭落叶松林17个科26种植物丛枝菌根(AM)定居情况和丛枝菌根真菌(AMF)多样性进行了调查。在26种植物根样中形成典型AM定居的有23种,占88.46%。菌根侵染率为0～78.7%,平均为21.69%。在26种植物根样中,4种植物形成重楼型(Par-is-type)AM,占15.38%;10种植物形成疆南星型(Arum-type)AM,占38.46%;9种植物形成中间型(Intermediatetype)AM,占34.62%,3种植物未能形成AM菌根,占11.54%。从23种植物根围土样中分离到50种AMF,分属于5个属,其中无梗囊霉属(Acaulospora)24种占48%;球囊霉属(Glomus)19种占38%;内养囊霉属(Entrophospora)5种占10%;原囊霉属(Archaeospora)1种占2%;巨孢囊霉属(Gigaspora)1种占2%。Acaulospora和Glomus2属真菌在大兴安岭落叶松林土壤中占绝对优势;尼氏无梗囊霉(A.nocolsonii)和幼套球囊霉(G.etunicatum)是优势种。植物根围土壤中AMF的孢子密度平均为68.94个.100g-1风干土;AM...     

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.     

荒漠油蒿（Artemisia ordosica）根围AM真菌分布与土壤酶活性

于2007年10月在油蒿(Artemisia ordosica)集中分布区选取4个典型样地,分别从0～10cm、10～20cm、20～30cm、30～40cm和40～50cm土层采集根围土样,分离其丛枝菌根(arbuscular mycorrhiza,AM)真菌并测定了3种土壤酶活性.结果表明,在各样地0～50cm土层中油蒿根系AM真菌菌丝定殖率均很高,有典型的泡囊与丛枝结构.菌丝定殖率与泡囊定殖率呈显著正相关,但菌丝、泡囊和丛枝定殖率与AM真菌孢子密度和土壤酶活性之间都无相关性;孢子密度在不同样地及采样深度间差异明显,与土壤有机质、速效磷和速效氮含量都呈一定正相关关系.孢子密度与脲酶和酸性磷酸酶活性有显著或极显著相关关系,与碱性磷酸酶活性之间的相关性受到土壤pH的显著影响.孢子密度的峰值出现在0～10cm表层土,并随土壤剖面深度增加而降低.土壤酶活性在土壤垂直剖面显示与孢子密度同样的规律.不同样地间AM真菌分布及油蒿根系定殖率的差异表明,油蒿与AM真菌之间有良好共生性,对维护荒漠生态环境系统结构的完整性具有重要意义.     

Ultrastructural localization of acid phosphatase in arbusculate coils of mycorrhizal Phoenix canariensis roots

Acid phosphatase (ACP) activity has been detected in roots of mycorrhizal and non-mycorrhizal Phoenix canariensis. This enzyme was ultrastructurally localized in arbusculate coils for the first time. This localization was carried out using a cerium-based method, which minimizes non-specific precipitation. The ACP was localized in inter- and intracellular hyphae, in the fungal cytoplasm as well as at the interface and the fungal cell wall and the periarbuscular membrane limiting it. The novel localization of an ACP in the arbuscular mycorrhizal (AM) interface of arbusculate coils suggests that this enzyme may be involved in the phosphorus efflux from the mycorrhizal fungus to the host. The results presented in this article indicate that the role played by ACP in AM symbiosis may be more important than was previously thought and that arbusculate coils are highly relevant when considering nutrient transfer through AM symbiosis.     

西北荒漠带细枝岩黄耆AM真菌与DSE真菌时空分布研究

为探明细枝岩黄耆Hedysarum scoparium根系丛枝菌根真菌（arbuscular mycorrhizae,AM）和深色有隔内生真菌（dark septate endophytes,DSE）定殖特征及其生态地理分布,充分理解菌根共生体在植物生长和植被恢复中的功能,本研究连续3年采集我国西北荒漠带不同样地细枝岩黄耆0-30cm土层土壤样品和根样,不同样地细枝岩黄耆均能被AM和DSE侵染,形成AM典型结构菌丝、泡囊和丛枝,DSE典型结构深色有隔菌丝和微菌核。细枝岩黄耆根系AM真菌定殖高于DSE定殖,但极端干旱条件下DSE定殖优于AM真菌定殖。AM真菌定殖率不同年际间表现为2016年最高,同一年份不同样地,除乌海、沙坡头样地,菌丝定殖率自东向西呈降低趋势。DSE真菌定殖率,在同一样地不同年份：乌海样地2016年定殖率显著高于2015年和2017年,在同一年份不同样地：2015年沙坡头样地DSE菌丝定殖率、总定殖率显著高于其他样地,而2016、2017年鄂尔多斯样地最高。NMDS分析结果显示,AM和DSE真菌定殖不同年际间差异显著。相关性分析表明,AM真菌定殖率与DSE定殖率显著正相关;土壤温度与AM定殖显著负相关;有机碳、速效磷、磷酸酶、湿度与AM定殖显著正相关;DSE菌丝定殖率、定殖强度仅与酸性磷酸酶正相关。本研究比较分析西北荒漠带不同样地AM和DSE定殖与土壤因子的相关性,有助于充分理解菌根共生体在植物生长和植被恢复中的功能和意义。     

Co-occurrence of Arum- and Paris-type morphologies of arbuscular mycorrhizae in cucumber and tomato

Colonization by arbuscular mycorrhizal (AM) fungi was investigated in cucumber (Cucumis sativus), tomato (Lycopersicon esculentum) and Clethra barbinervis (Ericales) grown in field-collected soil known from previous studies to generate Paris-type arbuscular mycorrhizae in C. barbinervis. Spores of Paraglomus, Acaulospora, Glomus, and Gigaspora were found in the soil. Formation of hyphal coils and arbusculate coils of Paris-type mycorrhizae and of arbuscules of Arum-type mycorrhizae in roots raised in this soil in the growth chamber were compared with the detection of DNA of AM fungi from the same root systems using Glomales-specific primers. Only Paris-type mycorrhizae with extensive arbusculate coils developed in C. barbinervis, but cucumber and tomato developed both Paris- and Arum-types in the same root systems. Glomaceae and Archaeosporaceae and/or Paraglomaceae were detected strongly in the DNA from both cucumber and tomato roots, in which Arum-type mycorrhizae were observed. In contrast, DNA of Glomaceae was detected more sparingly in C. barbinervis, in which Paris-type mycorrhizae dominated. Acaulosporaceae and Gigasporaceae were strongly detected in the DNA from both C. barbinervis and tomato, whereas they were more weakly detected in cucumber. These results indicate that the morphology of colonization is strongly influenced by the selection of fungi to colonize the host plant from among those in the soil environment.     

Light and Electron Microscopic Study on the Mycorrhizae of Sporophytes of Botrychium virginianum - Arbuscular Structure Resembling Fossil Forms

Abstract: The mycorrhizal structures of the sporophyte of the strictly protected fern Botrychium virginianum were studied by light and electron microscopy. The paper presented here is the first ultrastructural study of the mycorrhiza of the sporophyte of the genus Botrychium. The fungal colonization is restricted to two to three cell layers of the root cortex. The sometimes bifurcately ramifying hyphae form coils in the host cells. Single hypha arising from the coils penetrates into neighbouring cells and form arbuscules therein. The electron micrographs show high functional activity of the arbuscules, characterized by high glycogen-like content in the hyphae, a thin layer of filamentous matrix material in the interface between fungus and root cell, frequent cell organelles in the cytoplasm and lack of starch in the host cells containing arbuscules. The main distinctive feature of the mycorrhizae of Botrychium virginianum is the morphology of the arbuscules, which differs not only from the common fine-branched arbuscules, but also from the special structures reported previously from Ophioglossaceae. The robust, lobe-like branching arbuscules most resemble arbuscule structures found in Triassic fossils.     

内蒙古中西部草原主要植物的丛枝菌根及其结构类型研究

2002年6-9月,对内蒙古中西部草原建群植物及优势植物的丛枝菌根共生状况进行了调查。在观察的28科125种植物中,被丛枝菌根真菌侵染的植物有104种(占83．2％);在过去认为不能侵染的莎草科植物中发现卯穗苔草(Carex duriuscula)和黄囊苔草( C. korshinskyi)有侵染现象。在所调查的植物中,多年生草本和灌木类植物被丛枝菌根真菌侵染的比例较高,占被调查该类植物总数的90．4％;而一年生和二年生草本植物被侵染的比例仅为47．6％。本地区野生植物的丛枝菌根结构类型多数为Arum类型,占65．38％,尤其在百合科(Liliaceae)、菊科(Compositae)、豆科(Leguminosae)和蔷薇科(Rosaceae)植物中比例较高;而Paris类型仅有19．23％,多见于禾本科(Gramineae!)、唇形科(Labiatae)、桔梗科(Campanulaceae)和百合科。丛枝菌根的结构类型与植物的根系类型、生活型和菌根侵染率无关,而与植物所属的科属关系比较密切。     

Comparative study of mycorrhizal susceptibility and anatomy of four palm species

A morphological and anatomical study of the root systems of the palm species Brahea armata S. Watson, Chamaerops humilis L., Phoenix canariensis Chabaud and Phoenix dactylifera L. has been carried out to determine possible mycorrhizal colonization sites. Furthermore, the arbuscular mycorrhizal (AM) anatomical types formed by the four palm species in association with Glomus mosseae (Nicol. & Gerd.) Gerdemann & Trappe have been examined. The presence of a continuous sclerenchymatic ring in the outer cortex and aerenchyma in the inner cortex that are anatomical indicators of mycorrhizal nonsusceptibility in all four palm species is observed. The root systems of B. armata and C. humilis present only one group of third-order roots, while the third-order roots of P. canariensis and P. dactylifera may be divided into five different groups: short thick roots, mycorrhizal thickened roots, fine short roots, fine long roots, and pneumatorhizas. Third-order and some second-order roots of B. armata and C. humilis are susceptible to colonization by AM fungi, while only the mycorrhizal thickened roots form mycorrhizas with arbuscules in the Phoenix species. The root system of the Phoenix species also presents AM colonization in fine roots with only intraradical hyphae and spores, but without arbuscules, and pseudomantles of spores anchored in the pneumatorings of the second-order roots, which are described for the first time. The mycorrhizas formed by the four palm species are of an intermediate type, between the Arum and the Paris types, and are characterized by intercalary arbusculate coils and not only by intracellular but also by intercellular fungal growth. Our study suggests that a different degree of adaptation may exist among palm mycorrhizas toward the slow growth of palms and low spore numbers in the soil where they grow.     

Comparison of phosphatase localization in the intraradical hyphae of arbuscular mycorrhizal fungi,Glomus spp. and Gigaspora spp.

The localization of acid and alkaline phosphatases in the intraradical hyphae of the arbuscular mycorrhizal fungi, Glomus mosseae (Nicol. and Gerd.) Gerd. and Trappe (Gm), Gl. etunicatum Becker and Gerd. (Ge) and Gigaspora rosea Nicol. and Schenck (Gir) were compared. Marigold (Tagetes patula L.) and leek (Allium porrum L.) were inoculated with each of the three fungi. The mycorrhizal roots were harvested at 3, 4, 5 and 6 weeks after sowing (WAS), treated with a digestion solution containing cellulase and pectinase, and then stained for phosphatase activities at pH 5.0 and pH 8.5. The development of fungal structures in the host root was also examined. Gm formed fine-branched (mature) arbuscules only at the early phase of infection (3 to 4 WAS). Mature arbuscules of Ge and Gir were observed from the early phase (4 WAS) up to the end of experiment. At pH 5.0, the localization of the phosphatase activities of the three fungi were similar irrespective to host plant species. The activity appeared in mature arbuscules and intercellular hyphae, whereas the collapsed arbuscules were inactive. Ten millimolar NaF, an acid phosphatase inhibitor, inhibited the phosphatase activities of Gm and Ge but did not affect that of Gir. At pH 8.5, a difference among the fungal species was found in the localization of phosphatase activity while that between host species was not. The mature arbuscules were also the active sites in all three species. Only Gir showed the activity in the intercellular hyphae while the two Glomus spp. did not. Five millimolar EDTA inhibited the activity of Gir at pH 8.5 while the activities of Ge and Gm were not affected by either 5 mM EDTA or 10 mM KCN (both are alkaline phosphatase inhibitors).     

山西历山珍稀药用植物AM真菌资源与土壤因子的关系

以山西历山自然保护区暴马丁香、连香树、南方红豆杉和领春木4种珍稀药用植物为材料,研究其根际土壤中丛枝菌根真菌(AMF)的菌根结构类型和种属分布,同时分析土壤因子与其侵染率和孢子密度的关系。结果表明:(1)暴马丁香菌根类型为中间型(I-型),连香树和领春木为重楼型(P-型),南方红豆杉为疆南星型(A-型);4种植物根际共鉴定AMF 27种,分别隶属于球囊霉属(Glomus)、无梗囊霉属(Acaulospora)、盾巨孢囊霉属(Scutellos-pora)、多孢囊霉属(Diversispora)和原囊霉属(Archaeospora)5属,其中Glomus为优势属。(2)pH与暴马丁香和领春木的侵染率和孢子密度呈正相关,但与连香树和南方红豆杉呈负相关;速效磷与暴马丁香、南方红豆杉和领春木的侵染率和孢子密度呈正相关,但与连香树呈负相关;碱解氮、有机质与暴马丁香、连香树和领春木的侵染率和孢子密度呈正相关,但与南方红豆杉呈负相关。(3)4种药用植物的菌根侵染率主要受其根际土壤碱解氮的影响,而根际AMF孢子密度主要受根际土壤pH制约。可见,历山自然保护区内AMF资源丰富,多样性程度也较高;宿主植物不同,土壤因子对其侵染率和孢子密度的影响也不同。     

Expression patterns of defense-related genes in different types of arbuscular mycorrhizal development in wild-type and mycorrhiza-defective mutant tomato

The expression of defense-related genes was analyzed in the interactions of six arbuscular mycorrhizal (AM) fungi with the roots of wild-type tomato (Lycopersicon esculentum Mill.) cv. 76R and of the near-isogenic mycorrhiza-defective mutant rmc. Depending on the fungal species, wild-type tomato forms both major morphological AM types, Arum and Paris. The mutant rmc blocks the penetration of the root surface or invasion of the root cortex by most species of AM fungi, but one fungus has been shown to develop normal mycorrhizas. In the wild-type tomato, accumulation of mRNA representing a number of defense-related genes was low in Arum-type interactions, consistent with findings for this AM morphotype in other plant species. In contrast, Paris-type colonization, particularly by members of the family Gigasporaceae, was accompanied by a substantial transient increase in expression of some defense-related genes. However, the extent of root colonization did not differ significantly in the two wild-type AM morphotypes, suggesting that accumulation of defense gene products per se does not limit mycorrhiza development. In the mutant, interactions in which the fungus failed to penetrate the root lacked significant accumulation of defense gene mRNAs. However, phenotypes in which the fungus penetrated epidermal or hypodermal cells were associated with an enhanced and more prolonged gene expression. These results are discussed in relation to the mechanisms that may underlie the specificity of the interactions between AM fungi and the rmc mutant.     

丛枝菌根真菌在田间的分布特征、代谢活性及其对甘薯生长的影响

研究了大田条件下丛枝菌根(AM)真菌的分布特征、代谢活性及其对甘薯的生长效应.结果表明,接种Glomus intraradices 8周后,甘薯地上部干重,薯块鲜重和薯块个数均明显高于不接种对照;植株地上部和根系的吸磷量显著提高.与不接种对照相比,接种处理的甘薯菌根侵染率、甘薯根外菌丝密度以及甘薯根内菌丝的活性(根内菌丝碱性磷酸酶活性)显著提高.进一步分析甘薯根际不同方位上的菌丝分布,发现接种处理中平行于垄的方向的菌丝密度显著高于苗子下方的菌丝密度,而不接种处理的各个方向总菌丝密度无差异;活菌丝(具琥珀酸脱氢酶活性的菌丝)密度在各个方向的分布规律与总菌丝密度的分布规律一致.接种后根内菌丝活性的增强,根外活性菌丝密度的增加及其分布特征的改变,是甘薯产量增加的主要原因.     

Isolation and characterization of arbuscules from roots of an increased-arbuscule-forming mutant of Lotus japonicus

BACKGROUND AND AIMS: Previous methods for isolation of arbuscules from mycorrhizal roots are time-consuming, complex and expensive. Therefore, a simple, rapid and inexpensive method for the isolation of metabolically active arbuscules from plant root of an increased-arbuscule-forming mutant of Lotus japonicus (Ljsym78-2) is described. METHODS: Roots of the L. japonicus mutant plants Ljsym78-2 colonized by Glomus sp. were separated from soil, washed with water, immersed in CaSO(4) before being cut into 5-mm pieces and homogenized with a Waring blender at 6000 rpm for 30 s. The arbuscules were purified by separation from plant tissues with a 50-mum nylon mesh, finally collecting on a 30-mum nylon mesh. Enzyme histochemical staining showed that the collected arbuscules had succinate dehydrogenase, alkaline phosphatase and acid phosphatase activities. KEY RESULTS AND CONCLUSIONS: The enzymic activity of the arbuscules was not affected after the isolation process. The establishment of this simple, rapid and inexpensive method for the isolation of metabolically active arbuscules will be useful to clarify the biochemical processes occurring in nutrient exchange at the arbuscular interface.     

Association of highly and weakly mycorrhizal seedlings can promote the extra- and intraradical development of a common mycorrhizal network

Arbuscular mycorrhizal fungi (AMF) are key determinants of plant interactions in ecosystems. Through their effects on competition, they are regulators of the structure of communities. Conversely, the composition of plant assemblages may also influence the AMF colonization dynamics of plant species. Here, we tested under in vitro culture conditions the effects of Medicago truncatula, a highly mycorrhizal plant species, and Silene vulgaris, a weakly mycorrhizal plant species, grown single (monospecies treatments) or in combination (bispecies treatment) on the colonization dynamics of the AMF Rhizophagus irregularis MUCL 43194. The seedlings were placed in a pre-established hyphal network developing from a mature M. truncatula mycorrhizal donor plant. Extraradical mycelium (ERM) and root colonization parameters as well as root morphology were measured over a period of 12 days. An increased ERM length, total root colonization and proportion of arbuscules were noted in the bispecies treatment. Conversely, the bispecies treatment seemed to have no effect on root growth. This study also demonstrated the suitability of the in vitro culture system for studying the interactions between AMF and host plants grown as mono- and bispecies combinations.     

Structural differences in arbuscular mycorrhizal symbioses: more than 100 years after Gallaud,where next?

This review commemorates and examines the significance of the work of Isobel Gallaud more than 100 years ago that first established the existence of distinct structural classes (Arum-type and Paris-type) within arbuscular mycorrhizal (AM) symbioses. We add new information from recent publications to the previous data last collated 10 years ago to consider whether any patterns have emerged on the basis of different fungal morphology within plant species or families. We discuss: (1) possible control exerted by the fungus over AM morphology; (2) apparent lack of plant phylogenetic relationships between the classes; (3) functions of the interfaces in different structural classes in relation to nutrient transfer in particular; and (4) the occurrence of plants with both of the major classes, and with intermediate AM structures, in different plant habitats. We also give suggestions for future research to help remove uncertainties about the functional and ecological significance of differences in AM morphology. Lastly, we urge retention of the terms Arum- and Paris-type, which are now well recognised by those who study AM symbioses.