Aqueous biphasic system to extract arbuscular mycorrhizal spores from soils

The aqueous biphasic system (ABS) method using PEG and ammonium sulphate improved the recovery of spores from tropical soils as compared with the sucrose gradient method. The ABS was not affected by low temperatures. The optimal recovery of spores was achieved when 100% PEG and 50% ammonium sulphate concentration and PEG₈₀₀₀ instead of PEG₄₀₀₀ were used. Under these conditions an optimal interface formation which facilitates the localization and recovery of spores was obtained. Both phases of the ABS are aqueous and did not affect either the viability of spores or the level of root colonization reached by these spores. Soil composition did not affect the number of spores obtained by the ABS method either.     

丛枝菌根真菌对玉米生长生态效应的影响

1996～1998年试验研究了大田不施肥条件下3种泡囊丛枝菌根真菌Arbuscular mycorrhizal fungi (AMF),即 Glomus mosseae(G.m)、Glomus versifome(G.v)、Sclerocystis sinousa(S.s)对玉米生长的生态效应.结果表明,AMF处理的玉米根系活力为对照的2.32～3.05倍,玉米N素吸收比对照增加24.14%～56.65%,玉米P素吸收比对照增加41.14%～78.29%,促进玉米生长发育显著;在灭菌条件下G.m、G.v、S.s处理分别比对照增产17.63%、22.91%和10.80%;而不灭菌条件下3个处理分别比对照增产10.35%、20.56%和4.53%,同时显著提高玉米的水分供应状况.     

Response of nitrogen-transforming microorganisms to arbuscular mycorrhizal fungi

 We studied fluctuations in the numbers of autotrophic ammonium oxidizers, ammonifying microorganisms and denitrifying microorganisms in pot cultures of mycorrhizal and non-mycorrhizal maize. The populations were enumerated after 0, 15, 30, 45 and 60 days of plant growth. Two arbuscular mycorrhizal (AM) fungi belonging to different Glomus species were investigated. Pot cultures with AM-infected maize had significant quantitative and qualitative changes in the root-associated population of N-transforming bacteria compared with the non-mycorrhizal controls. The occurrence of autotrophic ammonium oxidizers in pot cultures of the AM fungi Glomus mosseae and G. fasciculatum was significantly higher than in non-mycorrhizal cultures throughout maize growth. The occurrence of these bacteria was delayed by 15 days in non-mycorrhizal as opposed to Glomus-colonized soil. Ammonifying and denitrifying bacterial populations were significantly decreased in the pot cultures of AM plants compared with the control. The distribution patterns of the physiological groups of bacteria tested were similar for both AM treatments but different from that of the non-mycorrhizal controls. Activity measurements expressed on a per cell basis showed changes with respect to the form of N in the mycorrhizal soil. G. fasciculatum was more active than G. mosseae during the earlier stages of plant growth. Received: 8 July 1997     

Characterization of glomalin as a hyphal wall component of arbuscular mycorrhizal fungi

Arbuscular mycorrhizal fungi (AMF) produce a protein, glomalin, quantified operationally in soils as glomalin-related soil protein (GRSP). GRSP concentrations in soil can range as high as several mg g⁻¹ soil, and GRSP is highly positively correlated with aggregate water stability. Given that AMF are obligate biotrophs (i.e. depending on host cells for their C supply), it is difficult to explain why apparently large amounts of glomalin would be produced and secreted actively into the soil, since the carbon could not be directly recaptured by the mycelium (and benefits to the AMF via increased soil structure would be diffuse and indirect). This apparent contradiction could be resolved by learning more about the pathway of delivery of glomalin into soil; namely, does this occur via secretion, or is glomalin tightly bound in the fungal walls and only released after hyphae are being degraded by the soil microbial community? In order to address this question, we grew the AMF Glomus intraradices in in vitro cultures and studied the release of glomalin from the mycelium and the accumulation of glomalin in the culture medium. Numerous protein-solubilizing treatments to release glomalin from the fungal mycelium were unsuccessful (including detergents, acid, base, solvents, and chaotropic agents), and the degree of harshness required to release the compound (autoclaving, enzymatic digestion) is consistent with the hypothesis that glomalin is tightly bound in hyphal and spore walls. Further, about 80% of glomalin (by weight) produced by the fungus was contained in hyphae and spores compared to that released into the culture medium, strongly suggesting that glomalin arrives mainly in soil via release from hyphae, and not primarily through secretion. These results point research on functions of glomalin and GRSP in a new direction, focusing on the contributions this protein makes to the living mycelium, rather than its role once it is released into the soil.     

Microbiota accompanying different arbuscular mycorrhizal fungal isolates influence soil aggregation

Arbuscular mycorrhizal fungi (AMF) are key components of ecosystems through their influence on plant communities and ecosystem processes. A major source of information regarding the importance of AMF species richness on process rates are mesocosm experiments using different levels of diversity of AMF as provided by single-species cultures of AMF. Since AMF inocula are generally made available in the form of non-sterile pot culture material, it is possible that AMF symbiosis-associated microbiota are at least partially responsible for some effects hitherto directly attributed to the AMF mycelium. Here, we provide evidence that microbiota associated with single-species cultures of AMF (after long-term pot culture enrichment of 7–8 years) can strongly affect the ecosystem process of soil aggregation. This effect occurred in an AMF isolate specific manner, but in the absence of live and active AMF mycelium. We additionally documented large differences in microbiota communities associated with the different AMF inocula (using PLFA analyses), suggesting that these differences were at least partly responsible for the observed changes in soil aggregation. This result points to AMF–microbiota interactions as a largely unexplored mechanism underlying soil aggregation (and potentially other ecosystem processes). We suggest that a reinterpretation of previous experiments using greenhouse-derived AMF cultures may be necessary, and the need to consider AMF symbiosis-associated microbiota in mechanistic studies of AMF and mycorrhizae in general is emphasized.     

Root herbivory by Tipula paludosa larvae increases colonization of Agrostis capillaris by arbuscular mycorrhizal fungi

The objective of this study was to determine if root-feeding by insect larvae affects subsequent colonization of roots by arbuscular mycorrhizal fungi. We investigated grazing by larvae of Tipula paludosa on colonization of Agrostis capillaris by two species of fungi, Glomus mosseae and G. intraradices. Host plants were subjected to 7 days of grazing only, continuous grazing for 42 days, or no herbivory. Those plants with no herbivory had significantly lower levels of colonization by arbuscules and hyphae compared to plants which were grazed for 7 or 42 days. The effect only occurred in the upper parts of the root system, where larvae were active. We suggest that this effect was most likely mediated by a change in quantity and composition of root exudates.     

Impact of conservation tillage and organic farming on the diversity of arbuscular mycorrhizal fungi

Communities of arbuscular mycorrhizal fungi (AMF) are strongly affected by land use intensity and soil type. The impact of tillage practices on AMF communities is still poorly understood, especially in organic farming systems. Our objective was to investigate the impact of soil cultivation on AMF communities in organically managed clay soils of a long-term field experiment located in the Sissle valley (Frick, Switzerland) where two different tillage (reduced and conventional mouldboard plough tillage) and two different types of fertilization (farmyard manure & slurry, or slurry only) have been applied since 2002. In addition, a permanent grassland and two conventionally managed croplands situated in the neighborhood of the experiment were analyzed as controls. Four different soil depths were studied including top-soils (0–10 and 10–20 cm) of different cultivation regimes and undisturbed sub-soils (20–30 and 30–40 cm). The fungi were directly isolated from field soil samples, and additionally spores were periodically collected from long-term trap culture (microcosm) systems. In total, >50,000 AMF spores were identified on the species level, and 53 AMF species were found, with 38 species in the permanent grassland, 33 each in the two reduced till organic farming systems, 28–33 in the regularly plowed organic farming systems, and 28–33 in the non-organic conventional farming systems. AMF spore density and species richness increased in the top-soils under reduced tillage as compared to the ploughed plots. In 10–20 cm also the Shannon–Weaver AMF diversity index was higher under reduced tillage than in the ploughed plots. Our study demonstrates that AMF communities in clay soils were affected by land use type, farming system, tillage as well as fertilization strategy and varying with soil depth. Several AMF indicator species especially for different land use types and tillage strategies were identified from the large data set.     

Soil type and land use intensity determine the composition of arbuscular mycorrhizal fungal communities

The objective of this study was to test whether soil types can be characterized by their arbuscular mycorrhizal fungal (AMF) communities. To answer this question, a well-defined study area in the temperate climatic zone of Central Europe was chosen with a large spectrum of soils and parent materials. Representative soil samples were taken from three soil types (Cambisol, Fluvisol and Leptosol) at in total 16 sites differing in agricultural land use intensity (9 grasslands and 7 arable lands). AMF spores were isolated and morphologically identified directly from field soils and after reproduction in trap cultures. AMF diversity and community composition strongly depended on soil type and land use intensity, and several AMF species were characteristic for a specific soil type or a specific land use type and hence had a specific niche. In contrast, other AM fungi could be considered as ‘generalists’ as they were present in each soil type investigated, irrespective of land use intensity. An estimated 53% of the 61 observed AMF species could be classified as ‘specialists’ as (almost) exclusively found in specific soil types and/or under specific land use intensities; 28% appeared to be ‘generalists’ and 19% could not be classified. Plant species compositions (either natural or planted) had only a subordinate influence on the AMF communities. In conclusion, land use intensity and soil type strongly affected AMF community composition as well as the presence and prevalence of many AM fungi. Future work should examine how the differences in AMF species compositions affect important ecosystem processes in different soils and to which extent the loss of specific groups of AM fungi affect soil quality.     

长期保护性耕作对丛枝菌根真菌多样性的影响

为了明确我国北方干旱地区长期保护性耕作以及深松对丛枝菌根真菌(AMF)多样性的影响,笔者于2014年在山西省临汾市连续22年实施保护性耕作的长期定位试验基地,针对免耕覆盖(NTS)、深松免耕覆盖(SNTS)及传统耕作(TT)3种处理方式,进行了不同耕作条件下土壤AMF物种丰度、孢子密度、Shannon多样性指数以及AMF侵染率等因素的比较研究。结果显示,长期保护性耕作(NTS和SNTS)共分离鉴定出AMF 7属9种,其中根孢囊霉属(Rhizophagus)和斗管囊霉属(Funneliformis)各2种,球囊霉属(Glomus)、近明球囊霉属(Claroideoglomus)、无梗囊霉属(Acaulospora)、硬囊霉属(Sclerocystis)和隔球囊霉属(Septoglomus)各1种;而传统耕作(TT)共分离鉴定出AMF 6属8种,没有检测到无梗囊霉属。NTS、SNTS和TT处理在不同土层的AMF优势种基本一致,0~40 cm土层为摩西斗管囊霉(Fu.mosseae)和变形球囊霉(G.versiforme),40~80 cm土层为摩西斗管囊霉、变形球囊霉和聚丛根孢囊霉(Rh.aggregatum),80~120 cm土层为聚丛根孢囊霉,120 cm土层以下只有NTS和SNTS处理中存在聚丛根孢囊霉,说明保护性耕作措施促进了AMF向土壤深层发展。NTS和SNTS处理在同一土层的AMF物种丰度、孢子密度和Shannon多样性指数均高于TT处理,SNTS处理高于NTS处理。同一耕作措施不同土层的AMF物种丰度、孢子密度和Shannon多样性指数均随土层加深而逐渐降低;NTS和SNTS处理在小麦各生育期的丛枝侵染率和孢子密度均高于TT处理;各处理在小麦拔节期的AMF侵染率最高,分别为14.9%、16.1%和10.6%,而在收获期的土壤孢子密度最高,分别为111.7个·(100g)~(-1)、125.0个·(100g)~(-1)和90.3个·(100g)~(-1)。研究认为,长期免耕覆盖、尤其深松免耕覆盖,提高了AMF多样性。该研究结果可为中国北方旱作农田生态系统中AMF自然潜力的充分发挥,以及保护性耕作技术的合理应用提供科学依据。     

Differential decomposition of arbuscular mycorrhizal fungal hyphae and glomalin

Arbuscular mycorrhizal fungi (AMF) are obligate symbionts of most higher plants. In addition to being a major component of soil microbial biomass, AMF hyphae produce glomalin, a recalcitrant glycoproteinaceous substance highly correlated with soil aggregate water stability. This study addresses the lack of knowledge concerning the decomposition of hyphae and glomalin. We used an experimental design that exploited the lack of saprobic capabilities of AMF hyphae by incubating field soil samples in the dark, and hence in the absence of plant or AMF hyphal growth. In 150 days, hyphal length decreased 60%, while glomalin, quantified by the Bradford protein assay, declined only 25%. Immuno-reactive glomalin decreased 46%. This study serves as a proof-of-concept for further examination of factors that influence decomposition of AMF hyphae using similar experimental designs.     

N,P Contribution and soil adaptability of four arbuscular mycorrhizal fungi

Abstract

A glasshouse study was conducted to investigate the symbiotic efficiency and soil adaptability of four AMF using glass-bead cultivation systems. The results showed that efficiency and adaptability of four fungi varied among three soils. Particularly, efficiency of BEG167 shifted from positive in Beijing soil to negative in Guangdong soil. Furthermore, BEG167 had high adaptability in all three soils. Intraspecific differences of BRG168 and BEG221 were found in efficiency and adaptability in three soils. Taking efficiency and adaptabilty into consideration, it was concluded that BEG167, BEG168 and BEG221 in Beijing soil, BEG151 in Hubei soil, and BEG151 and BEG168 in Guangdong soil were effective AMF for maize.     

Growth and interactions of arbuscular mycorrhizal fungi in soils from limestone and acid rock habitats

Arbuscular mycorrhizal (AM) development in different soil types, and the influence of AM fungal hyphae on their original soil were investigated. Plantago lanceolata, which can grow in soils of a very wide pH range, was grown in two closely related limestone soils and an acid soil from rock habitats. Plants were colonised by the indigenous AM fungal community. The use of compartmented systems allowed us to compare soil with and without mycorrhizal hyphae. Root colonisation of P. lanceolata was markedly higher in the limestone soils (30-60%) than in the acid soil (5-20%), both in the original habitat and in the experimental study. Growth of extraradical AM fungal hyphae was detected in the limestone soils, but not in the acid soil, using the signature fatty acid 16:1ω5 as biomass indicator. Analysis of signature fatty acids demonstrated an increased microbial biomass in the presence of AM fungal hyphae as judged for example from an increased amount of NLFA 16:0 with 30 nmol g⁻¹ in one of the limestone soils. Bacterial activity, but not soil phosphatase activity, was increased by around 25% in the presence of mycorrhizal hyphae in the first harvest of limestone soils. AM fungal hyphae can thus stimulate microorganisms. However, no effect of AM hyphae was observed on the soil pH or organic matter content in the limestone soils and the available P was not depleted.     

AM菌对三叶草吸收、累积重金属的影响

采用4室根箱培养系统,探讨了Cu、Zn、Pb、Cd 4种重金属复合污染土壤中,丛枝菌根菌对三叶草生长及吸收、累积重金属的作用,结果表明:重金属Cu 100mg/kg、Zn 600mg/kg、Pb 300mg/kg、Cd 10mg/kg的复合污染对三叶草生物量影响较小,但土壤重金属处理使丛枝菌根菌Glomus intraradices和Glomus caledonium对三叶草的侵染率分别降低53%和56%,菌种G.intraradice的菌丝密度降低73%;接种菌根真菌能明显减少重金属复合污染土壤中三叶草对Cu、Cd和Pb的吸收,并强化根系在限制重金属Pb和Cd向地上部运输中的作用,地上部Pb和Cd含量分别下降24.2%~55.3%和65%~97.9%,使三叶草地上部Cd和Pb含量均低于我国牧草重金属安全含量,提高了三叶草可食部分的质量;不同菌根真菌对三叶草吸收、累积及分配重金属的影响有明显差异,Glomus intraradices对减少三叶草对重金属的吸收及其在地上部可食部分的累积的作用大于Glomus caledonium。丛枝菌根菌对于强化三叶草根系对重金属的固持作用,调节生态系统中重金属的生物循环,减轻重金属对食物链的污染风险方面起着重要作用。     

VA菌根对土壤酸度的耐性

A 45－day greenhouse experiment was carried out to determine effect of vesicular-arbuscular（VA）my-corrhizal fungi on colonization rate,plant height,plant growth,hyphae lenth,total Al in the plants,ex-changeable Al in the soil and soil pH by comparison at soil pH 3.5,4.5 and 6.0 Plant mung bean(Phaseolus radiatus L.)and crotalaria(Crotalaria muronata Desv.) were grown with and without VA mycorrhizal fungi in pots with red soil,Ten VA mycorrhizal fungi strains were tested,including Glomus epigaeum(No.90001),Glomus caledonium(No.90036),Glomus mosseae(No.90107),Acaulospora soo.(No.34),Scutellopora het-erogama(No.36),Scutellospora calopsora(No.37),Glomus manihotis(No.38).Gigaspora spp.(No.47),Glomus manihotis(No.49),and Acaulospora spp.(No.53).Being the most tolerant to acidity,strain 34 and strain 38 showed quicker and higher-rated colnization without lagging,three to four times more in number of nodules,two to four times more in plant dry weighy,30% to 60% more in hyphae length,lower soil exchaneable Al,and higher soil pH than without VA mycorrhizal fungi(CK).Other strains also could improve plant growth and enhance plant tolerance to acidity,but their effects were not marked.This indicated that VA mycorrhizal fungi differed in the tolerance to soil acidity and so did their inoculation effects.In the experiment,acidic soil could be remedied by inoculation of promising VA mycorrhizal fungi tolerant of acidity.     

Global diversity and distribution of arbuscular mycorrhizal fungi

Arbuscular mycorrhizal (AM) fungi form associations with most land plants and can control carbon, nitrogen, and phosphorus cycling between above- and belowground components of ecosystems. Current estimates of AM fungal distributions are mainly inferred from the individual distributions of plant biomes, and climatic factors. However, dispersal limitation, local environmental conditions,and interactions among AM fungal taxa may also determine local diversity and global distributions. We assessed the relative importance of these potential controls by collecting 14,961 DNA sequences from 111 published studies and testing for relationships between AM fungal community composition and geography, environment, and plant biomes. Our results indicated that the global species richness of AM fungi was up to six times higher than previously estimated, largely owing to high beta diversity among sampling sites. Geographic distance, soil temperature and moisture, and plant community type were each significantly related to AM fungal community structure, but explained only a small amount of the observed variance. AM fungal species also tended to be phylogenetically clustered within sites, further suggesting that habitat filtering or dispersal limitation is a driver of AM fungal community assembly. Therefore, predicted shifts in climate and plant species distributions under global change may alter AM fungal communities.     

Influence of inoculation with arbuscular mycorrhizal fungi on stable isotopes of nitrogen in Phaseolus vulgaris

The interactions between Phaseolus vulgaris, Rhizobium spp. strains nodulating P. vulgaris, and arbuscular mycorrhizal (AM) fungi were assessed under greenhouse conditions in a nonsterilized Typic Haplustalf soil from Cauca, Colombia. Our results indicate a specific involvement of AM fungal species in nitrogen acquisition by the legume plants from symbiotic nitrogen fixation and from soil. A significant specific influence of inoculation with Glomus spp. on the ¹⁵N/¹⁴N ratio in plant shoots was dependent on the inoculated rhizobial strain, but AM fungal inoculation had no significant effect on shoot dry weight or nodule occupancy in the two different rhizobial strain treatments. The results imply that in low P soils the effects of an improved mycorrhizal symbiosis may include improved symbiotic N₂ fixation efficiency and/or improved soil N uptake. Received: 11 May 1996     

Glycosidation of apigenin results in a loss of its activity on different growth parameters of arbuscular mycorrhizal fungi from the genus Glomus and Gigaspora

The effect of different concentrations (0.5, 2 and 8 μM) of apigenin and its glycosidated form 5,7,4′-hydroxy flavone glycoside on arbuscular mycorrhizal (AM) fungal spore germination, hyphal growth, hyphal branching, the formation of entry points and root colonization of Gigaspora. rosea, Gi. margarita, Glomus mosseae and G. intraradices was tested. The lowest apigenin concentration (0.5 μM) nearly doubled hyphal branching, the formation of entry points and root colonization of all four tested fungi, whereas higher concentrations (2 and 8 μM) nearly doubled the hyphal growth of Gi. margarita, G. mosseae and G. intraradices. In none of the treatments with the apigenin-glycoside any effect on AM fungi could be observed. Our data show that apigenin exhibits an AM fungal genus and even species activity and we provide strong evidence that glycosidation results in a loss of its activity towards AM fungi.     

丛枝菌根与植物寄生性线虫相互作用及抗性机制

丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)能够与大多数陆地植物互惠共生,促进植物对养分的吸收,提高植物对各种生物和非生物胁迫的抗逆性,对植物健康生长有重要的作用。在土壤中丛枝菌根真菌与植物寄生性线虫共同依靠寄主植物根系完成生命循环,但二者对寄主植物作用完全相反,引起研究者广泛兴趣,成为菌根研究的热点和焦点之一。本文分析了丛植菌根真菌与植物寄生线虫的相互作用,并探讨了菌根提高植物对线虫抗性的可能机制:菌根真菌改善植物的生长和营养状况、改变植物根系形态结构、影响根系分泌物和根际微生物区系、诱导寄主植物产生防御反应等,旨在深入挖掘丛枝菌根真菌的生物学功能,进一步发挥其在农业生产中的应用潜力。     

Mycelium of arbuscular mycorrhizal fungi increases soil water repellency and is sufficient to maintain water-stable soil aggregates

Using an in vitro bioreactor system in which the arbuscular mycorrhizal (AM) fungus Glomus intraradices was grown in a soil devoid of detectable living microbes, we could show that the mycelium of this fungus contributed to the maintenance of water-stable soil aggregates and increased soil water repellency, as measured by water drop penetration time. This is to our knowledge the first demonstration of a causal link between AM fungal growth and water repellency of soil aggregates. Our results also place AM fungal contributions to soil aggregation on a firm mechanistic footing by showing that hyphae are sufficient to produce effects, in the absence of other soil biota, which have always been included in previous studies.     

Diversity of arbuscular mycorrhizal fungi in red tangerine (Citrus reticulata Blanco) rootstock rhizospheric soils from hillside citrus orchards

Root colonization, abundance of spores and hyphae, as well as species diversity of arbuscular mycorrhizal (AM) fungi were analyzed in citrus orchards along an altitudinal gradient. The citrus trees were heavily colonized (50.87–77.45%) by native AM fungi. In citrus orchards located at <600 m above sea level (asl), we recorded more extensive hyphal and arbuscular colonization, and higher spore and hyphal length density. AM fungal colonization, spore density, and hyphal length density were closely correlated with edaphic factors such as available phosphorus, pH, and organic matter. A total of 18 AM fungal species belonging to 3 different orders, Archaeosporales (1 species), Diversisporales (7 species) and Glomerales (10 species), were identified on the basis of spore morphological characteristics. In orchards located at higher altitudes (≥700 m asl), we observed a significant decrease in species richness and Shannon–Wiener index values. However, in all of the surveyed orchards, Glomus aggregatum, Funneliformis mosseae and Rhizophagus intraradices were the dominant species. Isolate frequency and relative abundance of AM fungi exhibited clearly distinct distribution patterns among taxonomic families. Canonical correspondence analysis revealed that the AM fungal community structure was significantly influenced by environmental factors, especially altitude, pH, soil moisture, and available nitrogen. Our data indicated that environmental factors are important in determining AM fungal root colonization, propagule numbers, and species diversity in citrus orchards.