Building a mycorrhizal cell: How to reach compatibility between plants and arbuscular mycorrhizal fungi

Abstract

Arbuscular mycorrhizal fungi occur throughout the majority of ecosystems supporting host plant nutrition. Recent findings describe the accommodation of the fungus by the root cell as a crucial step for compatibility between the partners. We discuss here the novel aspects of cellular plant-fungus interactions, with a particular attention to the interface compartment, the unique apoplastic space hosting intracellular fungal structures. The main features of arbuscular mycorrhizal colonization are examined and recent information in the field of plant and fungal cell responses during the establishment of the symbiosis is discussed. Differences between the colonization of root epidermal and cortical tissues are discussed, highlighting the growing interest in the role of epidermal cells during the first and decisive steps of the symbiosis. New approaches such as root organ cultures, in vivo observations, GFP tagging and mutant plant analysis are commented on and information from these is compared with that gained from more traditional methods. In particular, the use of plant mutants is depicted as a powerful tool for dissecting and understanding the genetic and cellular aspects of plant/fungus compatibility. Finally, perspectives in this field are outlined through the application of these approaches to the currently unanswered questions.     

Occurrence and coexistence of arbuscular mycorrhizal fungi and dark septate fungi in aquatic macrophytes in a tropical river-floodplain system

In this study, carried out in four water bodies in the Upper Paraná River floodplain, we assessed the occurrence of root colonization by arbuscular mycorrhizal fungi (AMF) and dark septate fungi (DSF), as well as the AMF species richness associated with 24 species of aquatic macrophytes belonging to different life forms. AMF were found in nine species of macrophytes and DSF in 16 species among the 24 investigated. AM colonization occurred mainly in eudicotiledons (five of the six species evaluated) and the Paris morphology was the most common type. Co-occurrence of AMF and DSF was observed in seven species of macrophytes (Commelinaceae sp. 1, Limnobium laevigatum (H.B.K. ex Willd) Heine, Hygrophila cf. costata, Myriophyllum brasiliense (Camb), Polygonum acuminatum Kunth, P. ferrugineum Wedd and P. stelligerum Cham). Four species of macrophytes (Pistia stratiotes L., Eichhornia crassipes (Mart.) Solms, Egeria najas Planch and Nymphaea amazonum Mart. & Zucc) were not colonized by any type of fungi. In total, 27 morphotypes of AMF were recorded, and spores occurred both in the rhizosphere of macrophytes whose roots were internally colonized by AMF and in non-colonized macrophytes. Acaulospora delicata, Acaulospora aff. laevis, Acaulospora longula, Glomus lamellosum, Glomus luteum and NID 1 (a non-identified species) were the most frequent species. Samples collected close to the roots of N. amazonum had the highest AMF richness (20 species), but this plant was not colonized by fungi. A species richness curve indicated that more root-associated fungi than reported here are likely present in this floodplain.     

Reproductive significance of feeding on saprobic and arbuscular mycorrhizal fungi by the collembolan, Folsomia candida

1. Collembolans have often been credited with negatively affecting arbuscular mycorrhizal (AM) symbioses, mainly by grazing and severing the associated external fungal network from host roots. However, most previous experiments were performed using relatively 'clean' systems where other, non-mycorrhizal, fungi were largely excluded. Yet, plant rhizospheres harbour a wide variety of highly palatable non-AM fungi, most of which have saprobic lifestyles.
2. In this study we isolated and cultured several rhizosphere fungi, and the collembolan , Folsomia candida , from the Long-Term Mycorrhiza Research Site, University of Guelph, Canada, to test the hypothesis that, given a choice, collembolans would prefer to feed on saprobic fungi and that such a choice is of adaptive significance to the animals.
3. A laboratory food preference experiment revealed that F. candida favours common saprobic fungi over a variety of AM fungi. Coincidentally, fecundity levels across two Folsomia generations were higher when animals fed exclusively on the preferred fungus, Alternaria alternata . When fed less palatable fungi, fecundity was greatly reduced; in fact animals from the F1 generation were unable to produce any eggs when placed on an exclusive diet of one of the following three AM fungi, Acaulospora spinosa, Scutellospora calospora and Gigaspora gigantea .
4. These results indicate that a strict diet of AM fungi by collembolans has reproductive consequences. Therefore, we propose that under natural conditions these animals spend more time feeding on common saprobic fungi rather than their AM counterparts. This suggests that previous 'clean' studies that investigated the interactions between collembolans and AM fungi may have reported exaggerated effects of animal grazing. The influence of collembolans on the functioning of AM symbioses, under more natural conditions, remains not well understood.     

Further root colonization by arbuscular mycorrhizal fungi in already mycorrhizal plants is suppressed after a critical level of root colonization

An established arbuscular mycorrhizal symbiosis suppresses further mycorrhization. It is not clear whether the observed suppressional effect is linked with the level of root colonization or not. In the present work we studied the effect of the degree of root colonization by the arbuscular mycorrhizal fungus Glomus mosseae on further root colonization by G. mosseae. At different time points barley plants grown in split-root compartments were pre-inoculated on one half of the split-root system with G. mosseae. Sequential inoculation resulted in different colonization levels. Thereafter, the second half of the split root system was inoculated. The results indicate an enhanced suppression of root colonization on the second side of the split-root system when colonization levels increased on the first side.     

Effects of arbuscular mycorrhizal fungi and soil developmental stages on herbaceous plants growing in the early stage of primary succession on Mount Fuji

A pot culture experiment was conducted to examine the effects of arbuscular mycorrhizal (AM) fungi and soil developmental stages on the growth and nutrient absorption of pioneer plants growing in the early stage of primary succession on Mt. Fuji. Four herbaceous plants, Polygonum cuspidatum (Polygonaceae), Miscanthus oligostachyus (Gramineae), Aster ageratoides var. ovatus (Compositae), and Hedysarum vicioides (Leguminosae), were grown from seed in soils collected from two different successional stages, bare ground and an herbaceous plant community. Spores of indigenous AM fungi collected from the herbaceous plant community were used as inoculum. The initial colonizer P. cuspidatum showed very low levels of AM colonization (<0.4%), whereas the average AM colonization levels of M. oligostachyus, A. ageratoides var. ovatus, and H. vicioides were within the range of 13–49%. AM fungi had positive effects on the growth and N acquisition of the leguminous species (H. vicioides) irrespective of soil developmental stages. In contrast, AM colonization did not increase the plant dry weight and N content of the non-leguminous species (P. cuspidatum, M. oligostachyus, and A. ageratoides var. ovatus) in both soil developmental stages. A positive effect of AM colonization on P content was observed in M. oligostachyus, A. ageratoides var. ovatus, and H. vicioides only in soil collected from the herbaceous plant community. P. cuspidatum showed no or a negative response to AM colonization in all cases. These results suggest that the effect of AM fungi on plant growth depends more on the plant species than soil developmental stages in the early stage of primary succession in this volcanic area.     

Biolistic transformation of arbuscular mycorrhizal fungi

Gene transfer systems have proved effective for the transformation of a range of organisms for both fundamental and applied studies. Biolistic transformation is a powerful method for the gene transfer into various organisms and tissues that have proved recalcitrant to more conventional means. For fungi, the biolistic approach is particularly effective where protoplasts are difficult to obtain and/or the organisms are difficult to culture. This is particularly applicable to arbuscular mycorrhizal (AM) fungi, being as they are obligate symbionts that can only be propagated in association with intact plants or root explants. Furthermore, these fungi are aseptate and protoplasts cannot be released. Recent advancements in gene transformation systems have enabled the use of biolistic technology to introduce foreign DNA linked to molecular markers into these fungi. In this review we discuss the development of transformation strategies for AM fungi by biolistics and highlight the areas of this technology which require further development for the stable transformation of these elusive organisms.     

Genetic evidence for the occurrence of assimilatory nitrate reductase in arbuscular mycorrhizal and other fungi

Using primers synthesized from two conserved regions and employing PCR, a DNA segment coding for part of the apoprotein of assimilatory nitrate reductase could be amplified from the fungi Aspergillus nidulans, Pythium intermedium, Phytophthora infestans, Phytophthora megasperma and Glomus D13. Sequencing of the amplificates as well as DNA hybridization revealed strong homologies with the nitrate reductase gene in all cases. The digoxigenin-labeled amplificate from Glomus hybridized with DNA isolated from Glomus spores. The data from these gene probing experiments are generally in accord with the published results from enzyme measurements. Thus assimilatory nitrate reductase occurs in saprophytic, parasitic as well as arbuscular mycorrhizal fungi. No amplificates with these primers were obtained with DNA isolated from Mucor mucedo and Saprolegnia ferax. Such results agree with the failure to detect nitrate assimilation physiologically in these two organisms.     

Improved aeroponic culture of inocula of arbuscular mycorrhizal fungi

We compared conventional atomizing disc aeroponic technology with the latest ultrasonic nebulizer technology for production of Glomus intraradices inocula. The piezo ceramic element technology used in the ultrasonic nebulizer employs high-frequency sound to nebulize nutrient solution into microdroplets 1 μm in diameter. Growth of pre-colonized arbuscular mycorrhizal (AM) roots of Sudan grass was achieved in both chambers used but both root growth and mycorrhization were significantly faster and more extensive in the ultrasonic nebulizer system than in the atomizing disc system. Shearing of the AM fungi (AMF) infected roots in both the systems did not reduce inoculum viability, as evident from the MPN data. However, sheared roots from the ultrasonic nebulizer system had significantly more infective propagules than those produced in the atomizing disc system. Thus, the latest ultra-sonic nebulizer aeroponic technology appears to be superior and an alternative to conventional atomizing disc or spray nozzle systems for the production of high-quality AMF inocula. These can be used in small doses to produce a large response, which is a prerequisite for commercialization of AMF technology. Accepted: 11 January 2000     

Multitrophic links between arbuscular mycorrhizal fungi and insect parasitoids

The effects of arbuscular mycorrhizal colonization of Leucanthemum vulgare on parasitism of a leaf‐mining insect was studied in a field and a laboratory experiment. In the field, parasitism of Chromatomyia syngenesiae by Diglyphus isaea was lower on mycorrhizal plants, compared with plants where the association was reduced. A laboratory experiment, in which L. vulgare was inoculated with three species of AM fungi, showed that the effects on parasitism rates were mycorrhizal species‐dependent. Some fungal combinations increased parasitism, some decreased it, while others had no effect. It is concluded that the most likely cause of these differences is plant size, with parasitoid searching efficiency being reduced on the larger plants, resulting from certain mycorrhizal species combinations. However, a mycorrhizal effect on herbivore‐produced plant volatiles cannot be ruled out.     

Interactions between Lotus japonicus genotypes and arbuscular mycorrhizal fungi

Abstract

Interactions between three genotypes (Ljsym 71-1, Ljsym 71-2 and Ljsym 72) of Lotus japoicus and one isolate from each of four species of arbuscular mycorrhizal fungi (Glomus sp. R-10, Glomus intraradices, Glomus etunicatum, and Gigaspora margarita) were investigated and compared with the wild-type ‘Gifu’ B-129. All the three genotypes showed no or defective internal colonization after inoculation with these AM fungi. In Ljsym72 mutant, the AM fungi produced deformed appressoria on the root surface, but failed to form any internal structures (internal hyphae, arbuscules and vesicles) except only in Glomus intraradices. The Ljsym71-1 and Ljsym71-2 mutants had more deformed appressoria and occasionally formed internal hyphae, arbuscules and vesicles, depending on AM fungi used. Wild-type ‘Gifu’ (nod⁺myc⁺) plants had typical colonization. The colonization of mutants by several fungi varied and provides a basis for studying recognition and compatibility between plants and mycorrhizal fungal species. These mutants also will be useful in studies of the genetics of the symbiosis between plant species and AM fungi.     

Detection of arbuscular mycorrhizal fungal spores in the air across different biomes and ecoregions

Aerial dispersal of fungal spores is common, but the role of wind and air movement in dispersal of spores of arbuscular mycorrhizal (AM) fungi is largely unknown. Several studies have examined the possibility of AM fungal spores being moved by wind vectors without observing spores taken from the air environment. For the first time this study observed the presence of AM fungal spores in the air. The frequency of AM fungal spores in the air was determined in six North American biomes composed of 18 ecoregions. Multiple samples were taken from both the air and the soil at each location. AM fungal spores were found in high abundance in the soil (hundreds of spores per gram of soil), however, they were rarely found in the air (most samples contained no AM fungal spores). Furthermore, only the Glomus morphotype was found in the air, whereas spores in the soil were taxomomically more diverse (Glomus, Acaulospora, Gigaspora, Scutellospora morphotypes were observed). The proportion of Glomus spores in the air relative to Glomus spores in the soil was highest in more arid systems, indicating that AM fungi may be more likely to be dispersed in the air in such systems. Nonetheless, the results indicate that the air is not likely a dominant mode of dispersal for AM fungi.     

中国大陆地区丛枝菌根真菌菌种资源的分离鉴定与形态学特征

【目的】分离收集保藏中国大陆各个地区不同生态环境的丛枝菌根真菌菌种资源,为丛枝菌根的研究提供资源、奠定基础。【方法】以高粱为宿主植物,采用诱导培养、单孢培养和扩繁培养分离土壤样品中的丛枝菌根真菌菌种并鉴定。【结果】从我国大陆的45个地区50余种宿主植物根区土壤中分离到丛枝菌根真菌135株,隶属于23个种;对各个菌株的形态特征进行了描述。【结论】我国蕴藏着丰富的丛枝菌根真菌菌种资源,文中描述的菌种资源是目前从我国大陆地区获得的种类和数量最多、覆盖范围最广的AM真菌菌种资源。     

入侵植物与丛枝菌根真菌的相互作用

入侵植物的入侵改变了入侵地生物群落的结构,导致生物多样性的丧失.丛枝菌根真菌(AMF)作为陆地生态系统中土壤微生物普遍的组成部分,它的种类和组成能够影响入侵植物的生长表现.这种真菌与寄主(入侵植物)特殊的关系也暗示了AMF能够影响入侵植物的入侵.反之,入侵植物的入侵同样也会影响AMF的群落结构和功能.本文在简要总结我国入侵植物种类及其危害的基础上,着重探讨了AMF与入侵植物入侵之间的关系,即AMF对入侵植物入侵过程中的作用、入侵植物入侵后如何影响AMF以及两者之间的相互作用机制.     

Response of strawberry to inoculation with arbuscular mycorrhizal fungi under very high soil phosphorus conditions

A field study was done to assess the potential benefit of arbuscular mycorrhizal (AM) inoculation of elite strawberry plants on plant multiplication, under typical strawberry nursery conditions and, in particular, high soil P fertility (Mehlich-3 extractible P=498 mg kg⁻¹). Commercially in vitro propagated elite plants of five cultivars (‘Chambly,’ ‘Glooscap,’ ‘Joliette,’ ‘Kent,’ and ‘Sweet Charlie’) were transplanted in noninoculated growth substrate or in substrate inoculated with Glomus intraradices or with a mixture of species (G. intraradices, Glomus mosseae, and Glomus etunicatum) at the acclimation stage and were grown for 6 weeks before transplantation in the field. We found that AM fungi can impact on plant productivity in a soil classified as excessively rich in P. Inoculated mother plants produced about 25% fewer daughter plants than the control in Chambly (P=0.03), and Glooscap produced about 50% more (P=0.008) daughter plants when inoculated with G. intraradices, while the productivity of other cultivars was not significantly decreased. Daughter plant shoot mass was not affected by treatments, but their roots had lower, higher, or similar mass, depending on the cultivar–inoculum combination. Root mass was unrelated to plant number. The average level of AM colonization of daughter plants produced by noninoculated mother plants did not exceed 2%, whereas plants produced from inoculated mothers had over 10% of their root length colonized 7 weeks after transplantation of mother plants and ∼6% after 14 weeks (harvest), suggesting that the AM fungi brought into the field by inoculated mother plants had established and spread up to the daughter plants. The host or nonhost nature of the crop species preceding strawberry plant production (barley or buckwheat) had no effect on soil mycorrhizal potential, on mother plant productivity, or on daughter plant mycorrhizal development. Thus, in soil excessively rich in P, inoculation may be the only option for management of the symbiosis.     

龙脑香科植物对丛枝菌根真菌的影响

在天然林地和温室盆栽条件下,比较研究了龙脑香科植物对丛枝菌根(Arbuscular mycorrhizas,AM)真菌孢子密度、相对多度、频度、属的组成、丰度和侵染状况等方面的影响.结果表明,用坡垒作盆栽寄主加富培养后,菌根侵染率、泡囊、丛枝和侵入点都低于原采样植物,以原坡垒土壤中栽植苗木的侵染率为最高,可达20.3%;而以望天树根围土壤栽植的苗木为最低,仅为10.6%;坡垒还不同程度地改变了原采样植物根围土壤中AM真菌孢子的密度、相对多度、频度、属的组成、丰度等.在4种土壤中,栽植坡垒苗木后,AM真菌的孢子密度都有不同程度的增长.采用与原采样相同种类的植物作为AM真菌加富培养的寄主更有利于促进AM真菌的生长发育、保持AM的多样性.     

Interactions between Glomus mosseae and arbuscular mycorrhizal sporocarp-associated saprophytic fungi

The saprophytic fungi Wardomyces inflatus (Marchal) Hennebert, Paecilomyces farinosus (Holm & Gray) A. H. S. Brown & G. Sm., Gliocladium roseum Bain., sterile dark mycelium (SDM-54), Trichoderma pseudokoningii Rifai and Trichoderma harzianum Rifai were isolated from sporocarps of Glomus mosseae. The effect of saprophytic fungi on G. mosseae spore germination was tested on water agar. Wardomyces inflatus decreased the percent germination of G. mosseae spores; G. roseum, T. pseudokoningii and T. harzianum had no effect on germination; and P. farinosus and SDM-54 increased the percentage of spore germination of G. mosseae after 4 d. Wardomyces inflatus significantly decreased hyphal length of spores which germinated, but no other saprophytic fungi affected hyphal growth. Trichoderma pseudokoningii, T. harzianum, P. farinosus and SDM-54 increased the number of auxiliary cells formed by G. mosseae. The effect of saprophytic fungi on arbuscular mycorrhizal colonization of soybean was studied in a greenhouse trial. The percentage of soybean root length colonized was decreased by W. inflatus, unaffected by SDM-54 and T. harzianum, and increased by P. farinosus. Gliocladium roseum decreased root length colonized when plants were 12 wk old, and T. pseudokoningii increased colonization of roots when plants were 4 wk old. Antagonistic, synergistic and neutral actions of G. mosseae upon the saprophytic fungi were observed. The population of T. harzianum decreased and the populations of T. pseudokoningii and SDM-54 increased in the presence of G. mosseae. Our results indicate a complex interaction between G. mosseae and associated saprophytic fungi.     

Genetic processes in arbuscular mycorrhizal fungi

Arbuscular mycorrhizal (AM) fungi (Glomeromycota) colonize roots of the majority of land plants and facilitate their mineral nutrient uptake. Consequently, AM fungi play an important role in terrestrial ecosystems and are becoming a component of sustainable land management practices. The absence of sexual reproductive structures in modern Glomeromycota combined with their long evolutionary history suggest that these fungi may represent an ancient asexual lineage of great potential interest to evolutionary biology. However, many aspects of basic AM fungal biology, including genome structure, within-individual genetic variation, and reproductive mode are poorly understood. These knowledge gaps hinder research on the mechanisms of AM fungal interactions with individual plants and plant communities, and utilization of AM fungi in agricultural practices. I present here the current state of research on the reproduction in AM fungi and indicate what new findings can be expected in the future.     

Red list plants: colonization by arbuscular mycorrhizal fungi and dark septate endophytes

Since information concerning the mycorrhization of endangered plants is of major importance for their potential re-establishment, we determined the mycorrhizal status of Serratula tinctoria (Asteraceae), Betonica officinalis (Lamiaceae), Drosera intermedia (Droseraceae) and Lycopodiella inundata (Lycopodiaceae), occurring at one of two wetland sites (fen meadow and peat bog), which differed in soil pH and available P levels. Root colonization by arbuscular mycorrhizal fungi (AMF) and dark septate endophytes (DSE) was quantified. Colonization by AMF appeared to be more frequent in the fen meadow than in the peat bog, and depended on the host plant. Roots of S. tinctoria and B. officinalis were well colonized by AMF in the fen meadow (35–55% root length) and both arbuscules and vesicles were observed to occur in spring as well as in autumn. In the peat bog, L. inundata showed a low level of root colonization in spring, when vesicles were found frequently but no arbuscules. In roots of D. intermedia from the peat bog, arbuscules and vesicles were observed, but AMF colonization was lower than in L. inundata. In contrast, the amount of AMF spores extracted from soil at the peat bog site was higher than from the fen meadow soil. Spore numbers did not differ between spring and autumn in the fen meadow, but they were higher in spring than in autumn in the peat bog. Acaulospora laevis or A. colossica and Glomus etunicatum were identified amongst the AMF spores extracted from soil at the two sites. S. tinctoria and B. officinalis roots were also regularly colonized by DSE (18–40% root length), while L. inundata was only rarely colonized and D. intermedia did not seem to be colonized by DSE at all.     

丛枝菌根真菌物种多样性研究进展

丛枝菌根真菌(Arbuscular mycorrhizal fungi,AMF)在不同生态系统均发挥至关重要的作用,研究其多样性能够为AMF物种资源的保护和利用提供科学依据。AMF不能被离体纯培养以及自身的高变异性等因素严重阻碍了对其进行深入研究,随着研究方法的不断改进,尤其是新一代测序技术的运用,极大加速了人们对AMF物种多样性的认识。本文主要从AMF分类系统、不同宿主植物和不同生境中的AMF物种多样性及AMF物种多样性研究方法(包括形态鉴定、Sanger测序和高通量测序)方面介绍AMF物种多样性研究进展,并且探讨AMF物种多样性研究中存在的主要问题,认为在今后AMF物种多样性研究中不仅要注重运用新的研究手段,还应该着重解决AMF不能离体纯培养的问题。