Glomalin: an arbuscular mycorrhizal fungal soil protein

Glomalin is abundant in soils and is closely correlated with aggregate water stability. Glomalin contains carbon and, hence, constitutes a non-trivial portion of the terrestrial carbon pool. Possibly far more importantly, however, stabilization of aggregates amplifies the role of glomalin in soils because carbonaceous compounds are protected from degradation inside of aggregates. Increased atmospheric CO₂ can lead to increased production of glomalin because of the symbiotic association that exists between plants and producers of glomalin, arbuscular mycorrhizal fungi (AMF). Glomalin concentrations in soils are influenced by management practices, for example, in agroecosystems, further highlighting the role of this protein in carbon storage. Glomalin is an unusual molecule that has proven difficult to analyze biochemically due to its recalcitrance and complexity. Future research will be directed towards the elucidation of its structure and controls on its production.     

Seasonal dynamics of arbuscular mycorrhizal fungal communities in roots in a seminatural grassland

Symbiotic arbuscular mycorrhizal fungi (AMF) have been shown to influence both the diversity and productivity of grassland plant communities. These effects have been postulated to depend on the differential effects of individual mycorrhizal taxa on different plant species; however, so far there are few detailed studies of the dynamics of AMF colonization of different plant species. In this study, we characterized the communities of AMF colonizing the roots of two plant species, Prunella vulgaris and Antennaria dioica, in a Swedish seminatural grassland at different times of the year. The AMF small subunit rRNA genes were subjected to PCR, cloning, sequencing, and phylogenetic analysis. Nineteen discrete sequence types belonging to Glomus groups A and B and to the genus Acaulospora were distinguished. No significant seasonal changes in the species compositions of the AMF communities as a whole were observed. However, the two plant species hosted significantly different AMF communities. P. vulgaris hosted a rich AMF community throughout the entire growing season. The presence of AMF in A. dioica decreased dramatically in autumn, while an increased presence of Ascomycetes species was detected.     

Polyphosphate dynamics in mycorrhizal roots during colonization of an arbuscular mycorrhizal fungus

Inorganic polyphosphate (poly P) has been considered to be a translocatable form of phosphate (Pi) in arbuscular mycorrhizal fungi (AMF). Here we examined time-course changes in poly P content during the AMF colonization process. Onion (Allium cepa) plants were cultured with or without inoculation with Gigaspora margarita for 2-8 wk with periodic sampling. Poly P in the extracts, purified through gel filtration, was quantified by the reverse reaction of polyphosphate kinase. The length of poly P in mycorrhizal roots appeared to be shorter than in extraradical hyphae or in spores of the AMF, indicating that AMF depolymerize poly P before providing Pi to the host. The poly P content increased as colonization proceeded, and was highly correlated with the weight of the colonized roots. These results support the model that AMF supply Pi to the host through the poly P pool, and that the poly P content of a mycorrhizal root can be a good indicator of the Pi-supplying activity of AMF.     

Gene copy number polymorphisms in an arbuscular mycorrhizal fungal population

Gene copy number polymorphism was studied in a population of the arbuscular mycorrhizal fungus Glomus intraradices by using a quantitative PCR approach on four different genomic regions. Variation in gene copy number was found for a pseudogene and for three ribosomal genes, providing conclusive evidence for a widespread occurrence of macromutational events in the population.     

一种改进的丛枝菌根染色方法

研究改进了Vierheilig等描述的AM菌根染色法:将根样于20%KOH溶液中60℃水浴透明40-120 min,5%醋酸酸化5min后,用5%醋酸墨水染色液(派克纯黑书写墨水Quink),于60℃水浴染色30 min,清水浸泡脱色(14h)后即可镜检。根皮层细胞内AM真菌的丛枝结构清晰可见,并且能够明确地分辨AM真菌与其它未知真菌。此外,Quink初染后,再经过SudanⅣ复染(60℃、60 min),70%乙醇脱色5min,暗隔真菌的透明菌丝内所积聚的脂类颗粒被SudanⅣ染上鲜红色,在复式显微镜下能够观察到此类透明菌丝在根皮层组织内的存在状况。采用甘油明胶为封固剂制片,根的染色效果可以保存长久。此项技术可以对同一种植物的多个根样进行同步的透明和染色处理,而且操作简便、低毒性、成本低廉、染色效果极佳,适用于野生和栽培草本植物AM菌根的染色和制片观察。     

Dating in the dark: how roots respond to fungal signals to establish arbuscular mycorrhizal symbiosis

The arbuscular mycorrhizal symbiosis that involves most plants and Glomeromycota fungi is the result of a complex exchange of molecular information, which commences before the partners are in physical contact. On the one hand, plants release soluble factors, including strigolactones that activate both the metabolism and branching of the fungal partners. On the other hand, fungi use compounds that trigger the signaling transduction pathways that are required for the symbiotic modus of plant cells. Here we describe some of the recent discoveries regarding the fungal molecules involved in rhizospheric conversation, and the way in which they are perceived by their hosts. We conclude that similar signaling molecules may have different meanings, depending on the context. However, at the end, specificity must be maintained to ensure appropriate partners enter symbiosis.     

中国丛枝菌根真菌分子多样性

【背景】随着分子生物学技术的发展,通过分子生物学方法鉴定丛枝菌根真菌(Arbuscular mycorrhizal fungi,AMF)多样性的研究越来越多。目前,AMF在全球的分子多样性及其分布已引起关注,但在我国关于AMF分子多样性的研究进展状况尚未见报道。【目的】探究我国AMF分子多样性及其分布,研究不同气候区域以及不同生态系统中AMF分子多样性并揭示其分布规律,为我国AMF在分子水平上的研究提供基础数据。【方法】利用Davison于2015年发表于"Science"上的一篇有关全球AMF分子多样性的数据库,选用中国AMF多样性的数据,并补充了近年来已发表的该数据库中未涉及的数据,建立中国AMF分子多样性的新数据库。【结果】共得到145个分子生物学鉴定出的虚拟种(Virtualtaxa,VT),隶属于8科12属。对于不同生态系统而言,草地生态系统中AMF的分子多样性最高,占到总VT数的60.7%,人为生态系统和森林生态系统也分别达到55.2%和43.4%。同样,AMF的VT数也随气候区域的不同而有所变化,温带气候区域所占比例最高(64.1%),亚热带气候区域次之(60.7%),极北气候区域最少(20.7%)。而对于根系和土壤样品中AMF的分子多样性而言,根系样品中发现的AMF分子多样性(80.0%)高于土壤中AMF的分子多样性(48.3%)。进一步对不同生态系统、气候区域和样品来源分别在属的水平上进行研究,同样也存在一定的差异。【结论】我国具有丰富的AMF分子多样性,且不同气候区域、生态系统以及样品来源都影响着AMF的分子多样性及其分布。     

丛枝菌根真菌种群的孢子季相动态研究

以草坪为研究对象,研究草坪土壤中丛枝菌根真菌种群的孢子组成、孢子密度、物种丰富度、多样性及其季相变化规律。结果表明,丛枝菌根真菌孢子密度、物种丰富度和多样性指数在一年内随季节变化表现出一定的季相变化规律,三者均在冬季达到最高,在时间节律上与植物群落季相变化不同步;同时分析了气候因素(平均温度、降雨量和日照时间等)对丛枝菌根真菌的影响。结果表明,气候因素对孢子密度、物种丰富度和多样性指数均有显著影响。     

丛枝菌根真菌繁殖体的高效扩繁

【背景】丛枝菌根(arbuscular mycorrhiza, AM)真菌能够和大多数植物形成互利共生体系,以促进植物生长、提高抗逆能力,在生产中具有重要作用,但AM真菌的繁殖技术限制了其应用。【目的】构建AM真菌的高效繁殖体系。【方法】于温室盆栽条件下,将根内根孢囊霉(Rhizophagus intraradice)接种于由3种寄主植物高粱(Sorghum bicolor)、玉米(Zea mays)、红三叶草(Trifolium repens)与5种培养基质(沸石、河砂、草炭、珍珠岩和蛭石)构建的4种繁殖体系中进行培养。研究不同繁殖体系对根内根孢囊霉侵染程度、产孢量的影响;然后利用高粱接种扩繁的菌剂进行AM真菌侵染能力的测定以验证其扩繁效果;最后基于筛选出的最优扩繁条件探讨对其他种类AM真菌摩西斗管囊霉(Funneliformis mosseae)、幼套近明球囊霉(Clariodeoglous etunicatum)、地表多样孢囊霉(Diversispora versiformis)和脆无梗囊霉(Acaulospora delicate)的扩繁效果。【结果】基质为河砂+蛭石+草炭(体...     

丛枝菌根真菌群落沿高寒草原海拔梯度的变化特征

基于丛枝菌根(Arbuscular mycorrhizal,AM)真菌孢子形态学的鉴定,对沿不同海拔(4584、4628、4744、4880、4956 m)梯度采集的高寒草原建群植物根际土壤样品进行了分析。结果表明,高寒草原AM真菌属、种构成均较简单,Acaulospora、Claroideoglomus、Funneliformis、Glomus属见于各海拔梯度,海拔4744 m地带未见Pacispora属,海拔4744、4956 m地带无Scutellospora属分布,Rhizophagus属仅见于海拔4584 m地带。随海拔上升,AM真菌种数、物种丰度均呈显著下降;海拔4584—4880 m范围Shannon-Weiner指数(H)无显著差异,但在最高海拔时显著下降;优势种种数及所占比例与海拔梯度则呈显著正相关(Funneliformis geosporum、Claroideoglomus claroideum为不同海拔梯度优势种);沿海拔梯度,孢子密度基本呈单峰分布格局,峰值出现在海拔4744 m地带;海拔梯度对菌根侵染效应影响显著,菌根侵染率、侵染强度和丛枝丰度随海拔上升均呈显著下降趋势;不同海拔梯度高寒草原AM真菌群落相似度(Sorensen相似性系数0.821—0.969)较高,并在总体上表现出随海拔梯度增大而降低的趋势。土壤pH值、有效磷、有机碳、海拔对AM真菌的群落分布均产生显著影响,尤以海拔的影响最为显著。研究结果对预测高寒草原微生物的作用与影响,以及高寒草原环境对全球变化的响应等提供了理论依据。     

Unique arbuscular mycorrhizal fungal communities uncovered in date palm plantations and surrounding desert habitats of Southern Arabia

The main objective of this study was to shed light on the previously unknown arbuscular mycorrhizal fungal (AMF) communities in Southern Arabia. We explored AMF communities in two date palm (Phoenix dactylifera) plantations and the natural vegetation of their surrounding arid habitats. The plantations were managed traditionally in an oasis and according to conventional guidelines at an experimental station. Based on spore morphotyping, the AMF communities under the date palms appeared to be quite diverse at both plantations and more similar to each other than to the communities under the ruderal plant, Polygala erioptera, growing at the experimental station on the dry strip between the palm trees, and to the communities uncovered under the native vegetation (Zygophyllum hamiense, Salvadora persica, Prosopis cineraria, inter-plant area) of adjacent undisturbed arid habitat. AMF spore abundance and species richness were higher under date palms than under the ruderal and native plants. Sampling in a remote sand dune area under Heliotropium kotschyi yielded only two AMF morphospecies and only after trap culturing. Overall, 25 AMF morphospecies were detected encompassing all study habitats. Eighteen belonged to the genus Glomus including four undescribed species. Glomus sinuosum, a species typically found in undisturbed habitats, was the most frequently occurring morphospecies under the date palms. Using molecular tools, it was also found as a phylogenetic taxon associated with date palm roots. These roots were associated with nine phylogenetic taxa, among them eight from Glomus group A, but the majority could not be assigned to known morphospecies or to environmental sequences in public databases. Some phylogenetic taxa seemed to be site specific. Despite the use of group-specific primers and efficient trapping systems with a bait plant consortium, surprisingly, two of the globally most frequently found species, Glomus intraradices and Glomus mosseae, were not detected neither as phylogenetic taxa in the date palm roots nor as spores under the date palms, the intermediate ruderal plant, or the surrounding natural vegetation. The results highlight the uniqueness of AMF communities inhabiting these diverse habitats exposed to the harsh climatic conditions of Southern Arabia.     

"Chromoplast" development in arbuscular mycorrhizal roots

The accumulation of apocarotenoids in arbuscular mycorrhizal (AM) roots suggests a dramatic reorganization of the plastids responsible for the biosynthesis of these compounds. This review describes the cytological and biochemical characterization of this phenomenon. The results presented suggest that plastids are key organelles for the establishment of the symbiotic interface of the AM symbiosis. In addition, a complex interplay of various plant cell components during the different functional phases of this interface is suggested. Arbuscule degradation appears to be of particular interest, as it correlates with the formation of the most extensive plastid structures and with apocarotenoid accumulation.     

Accumulation of reactive oxygen species in arbuscular mycorrhizal roots

We investigated the accumulation of reactive oxygen species (ROS) in arbuscular mycorrhizal (AM) roots from Medicago truncatula, Zea mays and Nicotiana tabacum using three independent staining techniques. Colonized root cortical cells and the symbiotic fungal partner were observed to be involved in the production of ROS. Extraradical hyphae and spores from Glomus intraradices accumulated small levels of ROS within their cell wall and produced ROS within the cytoplasm in response to stress. Within AM roots, we observed a certain correlation of arbuscular senescence and H₂O₂ accumulation after staining by diaminobenzidine (DAB) and a more general accumulation of ROS close to fungal structures when using dihydrorhodamine 123 (DHR 123) for staining. According to electron microscopical analysis of AM roots from Z. mays after staining by CeCl₃, intracellular accumulation of H₂O₂ was observed in the plant cytoplasm close to intact and collapsing fungal structures, whereas intercellular H₂O₂ was located on the surface of fungal hyphae. These characteristics of ROS accumulation in AM roots suggest similarities to ROS accumulation during the senescence of legume root nodules.     

Potato-associated arbuscular mycorrhizal fungal communities in the Peruvian Andes

The world's fourth largest food crop, potato, originates in the Andes. Here, the community composition of arbuscular mycorrhizal fungi (AMF) associated with potato in Andean ecosystems is described for the first time. AMF were studied in potato roots and rhizosphere soil at four different altitudes from 2,658 to 4,075 m above mean sea level (mamsl) and in three plant growth stages (emergence, flowering, and senescence). AMF species were distinguished by sequencing an approx. 1,500 bp nuclear rDNA region. Twenty species of AMF were identified, of which 12 came from potato roots and 15 from rhizosphere soil. Seven species were found in both roots and soil. Interestingly, altitude affected species composition with the highest altitude exhibiting the greatest species diversity. The three most common colonizers of potato roots detected were Funneliformis mosseae, an unknown Claroideoglomus sp., and Rhizophagus irregularis. Notably, the potato-associated AMF diversity observed in this Andean region is much higher than that reported for potato in other ecosystems. Potato plants were colonized by diverse species from 8 of the 11 Glomeromycota families. Identification of the AMF species is important for their potential use in sustainable management practices to improve potato production in the Andean region.     

Changes in arbuscular mycorrhizal fungal communities during invasion by an exotic invasive plant

Exotic invasive plants can show strong plant–soil feedback responses, but little is known about time scales for significant changes in soil microbial communities to occur after invasion. Previous work has suggested that plant invasions can modify arbuscular mycorrhizal (AM) fungal community structure. However, there is a lack of understanding about how long it takes for these changes to develop. To test this we investigated temporal changes in AM fungal communities colonising the invasive plant Vincetoxicum rossicum (Apocynaceae). We hypothesised that AM fungal community structure would change in a particular direction during the invasion process. We collected soil from two sites with a long history of invasion by this plant, with each site having paired invaded and uninvaded plots. Soil from these plots was used in a glasshouse experiment to characterise AM fungal community structure in the roots of V. rossicum at different times throughout a simulated growing season. AM fungal community structure differed between invaded and uninvaded plots. However, contrasting with our hypothesis, AM fungal communities colonising V. rossicum growing in soil from uninvaded plots did not change towards those in plants growing in previously invaded soil. Our data suggest that changes to AM fungal communities in the presence of V. rossicum require longer than the first growing season after establishment to develop.