Effects of metal phytoextraction practices on the indigenous community of arbuscular mycorrhizal fungi at a metal-contaminated landfill

Phytoextraction involves use of plants to remove toxic metals from soil. We examined the effects of phytoextraction practices with three plant species (Silene vulgaris, Thlaspi caerulescens, and Zea mays) and a factorial variation of soil amendments (either an ammonium or nitrate source of nitrogen and the presence or absence of an elemental sulfur supplement) on arbuscular mycorrhizal (AM) fungi (Glomales, Zygomycetes) at a moderately metal-contaminated landfill located in St. Paul, Minn. Specifically, we tested whether the applied treatments affected the density of glomalean spores and AM root colonization in maize. Glomalean fungi from the landfill were grouped into two morphotypes characterized by either light-colored spores (LCS) or dark-colored spores (DCS). Dominant species of the LCS morphotype were Glomus mosseae and an unidentified Glomus sp., whereas the DCS morphotype was dominated by Glomus constrictum. The density of spores of the LCS morphotype from the phytoremediated area was lower than the density of these spores in the untreated landfill soil. Within the experimental area, spore density of the LCS morphotype in the rhizosphere of mycorrhizal maize was significantly higher than in rhizospheres of nonmycorrhizal S. vulgaris or T. caerulescens. Sulfur supplement increased vesicular root colonization in maize and exerted a negative effect on spore density in maize rhizosphere. We conclude that phytoextraction practices, e.g., the choice of plant species and soil amendments, may have a great impact on the quantity and species composition of glomalean propagules as well as on mycorrhiza functioning during long-term metal-remediation treatments.     

西双版纳热带雨林中丛枝菌根真菌的初步研究*

对西双版纳热带雨林中30个科的42种植物根系的丛枝菌根真菌定居情况进行了调查,并从这些植物的根际土壤中分离鉴定了分属于无梗囊霉属(Acaulospora)、球囊霉属(Glomus)和硬囊霉属(Sclerocystis)的25种丛枝菌根真菌。对热带雨林土壤中丛枝菌根真菌的孢子密度（spore density）、物种丰富度（species richness）以及已鉴定种的出现频率进行统计分析发现：热带雨林土壤中丛枝菌根真菌的孢子密度在每100g土壤116~1560个之间,平均478个;物种丰富度在2~7之间,平均为4.5;无梗囊霉属和球囊霉属真菌是热带雨林土壤中丛枝菌根真菌的优势类群。     

Spore production and mycorrhizal development in various tropical crop hosts infected with Glomus clarum

Plant growth, mycorrhizal development and vesicular arbuscular spore production were examined in five tropical crop host species inoculated with Glomus clarum and grown in a glasshouse. In one of the two experiments, sequential harvests of maize, sorghum and chickpea were made in order to study spore production in relation to plant growth and mycorrhizal development. Spore numbers in each of these hosts increased at a fairly constant rate until maximum plant dry weight, when spore production ceased. Sorghum and maize produced considerably more spores than chickpea, with spore numbers being closely correlated with mycorrhizal root length. In the second experiment, Glomus clarum was cultured on each of maize, millet, sorghum, groundnut and chickpea for three consecutive generations before cross-inoculation of the spores from each host onto all five hosts. Sporulation with respect to host size was generally greatest when the inoculum used to infect a host had been produced on that host. The growth-promoting effects of the fungus were not influenced by the source of the inoculum. More spores were produced on the cereals than the legumes. Differences in spore numbers amongst hosts and plant generations were apparently influenced mainly by infected root length and by the growth period.     

Effect of edaphic factors and seasonal variation on spore density and root colonization of arbuscular mycorrhizal fungi in sugarcane fields

Sugarcane fields in 14 different study sites were analyzed for the presence of different arbuscular mycorrhizal fungal (AMF) spores. A total of 23 AMF species representing four genera were identified, among which Glomus fasciculatum and G. mosseae were the dominant species. The mean spore density in the root-zone soils of sugarcane plants varied from 119 to 583 per 100 g of soil, and the mean percentage root colonization varied from 60 to 89 %. A study of the effect of edaphic factors on AM spore density and percentage root colonization revealed a positive correlation between pH and AMF spore density and root colonization and a negative correlation between electrical conductivity, nitrogen, and phosphorus. A positive correlation was observed between AMF spore density and root colonization. Season was also found to play a vital role in determining AMF spore density and percentage root colonization, with high spore density and root colonization observed during the summer season and lower spore densities and root colonization during the winter season.     

龙脑香科植物AM真菌的初步研究

对我国云南和海南龙脑香科植物的天然原始林和人工林AM真菌资源进行了调查,对从46处龙脑香林采回的19种龙脑香科树种的根系和根际土壤样品,分别进行显微观察和分离AM真菌的孢子,发现在所调查的19个树种中,所有树种的根系都有不同程度的AM真菌的侵染,高者可达50%,平均为21%,其中在大多数根样中发现有泡囊生成;对土壤样品的分析,分离到大量的孢子,其孢子密度从7~132个/20ml土样不等,平均为58个。对所分离到的孢子进行鉴定,发现无梗囊霉属和球囊霉属的孢子占大多数,其频度和相对多度分别为33.25和46.89;18.04和30.80。研究结果表明,龙脑香科不仅仅能够形成外生菌根,而且也能象大多数热带林木一样能够广泛的形成丛枝菌根。     

Distribution of vesicular-arbuscular mycorrhizal fungi in the natural ecosystem

Natural occurrence of vesicular-arbuscular mycorrhizal (VAM) fungi in Haryana soils showed that VAM sporulation was more intensive in the rhizosphere of nonlegumes than of legumes. Maximum number of spores (342 spores per 50 g of soil) was observed in the rhizosphere of mustard, followed by chickpea, wheat, pearl millet and pigeonpea. Four VAM generaviz. Glomus, Gigaspora, Sclerocystis andAcaulospora, were present there. Soil pH, total soil P, available P, type of soil, soil moisture and cropping season all variables influenced the VA mycorrhizal population in the natural ecosystem. Numbers of VAM spores highly correlated with the presence of total soil P and soil pH indirectly affected the VAM population through the total soil P. The spore population was abundant in sandy soils as compared to loamy sands. Drier soils had higher number of VAM spores. In summer, the VAM population in soil was less as compared to winter season.     

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。     

Glomus intraradices dominates arbuscular mycorrhizal communities in a heavy textured agricultural soil

Arbuscular mycorrhizal fungal (AMF) spore communities were surveyed in a long-term field fertilization experiment in Switzerland, where different amounts of phosphorus (P) were applied to soil. Plots receiving no P as well as plots systematically fertilized in excess to plant needs for 31 years were used to test the hypothesis that application of P fertilizer changes the composition and diversity of AMF communities. AMF spores were isolated from the field soil, identified, and counted so as to quantify the effect of P fertilization on AMF spore density, composition, and diversity. Trap cultures were established from field soil with four host plants (sunflower, leek, maize, and Crotalaria grahamiana), and the spore communities were then analyzed in substrate samples from the pots. Altogether, nine AMF species were detected in the soil. No evidence has been acquired for effect of P fertilization on spore density, composition, and diversity of AMF in both the field soil and in trap cultures. On the other hand, we observed strong effect of crop plant species on spore densities in the soil, the values being lowest under rapeseed and highest under Phacelia tanacetifolia covercrop. The identity of plant species in trap pots also significantly affected composition and diversity of associated AMF communities, probably due to preferential establishment of symbiosis between certain plant and AMF species. AMF spore communities under mycorrhizal host plants (wheat and Phacelia in the fields and four host plant species in trap pots) were dominated by a single AMF species, Glomus intraradices. This resulted in exceptionally low AMF spore diversity that seems to be linked to high clay content of the soil.Electronic supplementary material Supplementary material is available for this article at and accessible for authorised users.     

Interaction of vascular plants and vesicular-arbuscular mycorrhizal fungi across a soil moisture-nutrient gradient

Summary Abundance and distribution of vascular plants and vesicular-arbuscular mycorrhizal (VAM) fungi across a soil moisture-nutrient gradient were studied at a single site. Vegetation on the site varied from a dry mesic paririe dominated by little bluestem (Schizachyrium scoparium) to emergent aquatic vegetation dominated by cattail (Typha latifolia) and water smartweed (Polygonum hydropiperoides). Plant cover, VAM spore abundance, plant species richness, and number of VAM fungi represented as spores, had significant positive correlations with each other and with percent organic matter. The plant and VAM spore variables had significant negative correlations with soil pH and available Ca, Mg, P and gravimetric soil moisture. Using stepwise multiple regression, Ca was found to be the best predictor of spore abundance. Test for association between plant species and VAM fungal spores indicated that the spores of Glomus caledonium are associated with plants from dry, nutrient poor sites and spores of gigaspora gigantea are positively associated with plants occurring on the wet, relatively nutrient rich sites. Glomus fasciculatum was the most abundant and widely distributed VAM fungus and it had more positive associations with endophyte hosts than the other VAM fungi. We found no relationship between beta niche breadth of plant species and the presence or absence of mycorrhizal infection. However, our data suggest that some plant species may vary with respect to their infection status depending upon soil moisture conditions that may fluctuate seasonally or annually to favor or hinder VAM associations.     

Status of arbuscular mycorrhizal fungi (AMF) in the Sundarbans of India in relation to tidal inundation and chemical properties of soil

The arbuscular mycorrhizal status of fifteen mangroves and one mangrove associate was investigated from 27 sites of three inundation types namely, diurnal, usual springtide and summer springtide. Roots and rhizospheric soil samples were analysed for spore density, frequency of mycorrhizal colonization and some chemical characteristics of soil. Relative abundance, frequency and spore richness of AMF were assessed at each inundation type. All the plant species except Avicennia alba exhibited mycorrhizal colonization. The study demonstrated that mycorrhizal colonization and spore density were more influenced by host plant species than tidal inundation. Forty four AMF species belonging to six genera, namely Acaulospora, Entrophospora, Gigaspora, Glomus, Sclerocystis and Scutellospora, were recorded. Glomus mosseae exhibited highest frequency at all the inundation types; Glomus fistulosum, Sclerocystis coremioides and Glomus mosseae showed highest relative abundance at sites inundated by usual springtides, summer springtides and diurnal tides, respectively. Spore richness of AMF was of the order usual springtide > diurnal > summer springtide inundated sites. The mean spore richness was 3.27. Diurnally inundated sites had the lowest concentrations of salinity, available phosphorus, exchangeable potassium, sodium and magnesium. Statistical analyses indicated that mycorrhizal frequency and AMF spore richness were significantly negatively correlated to soil salinity. Spore richness was also significantly negatively correlated to available phosphorus. The soil parameters of the usual springtide inundated sites appeared to be favourable for the existence of maximum number of AMF. Glomus mosseae was the predominant species in terms of frequency in the soils of the Sundarbans.     

Identification of a Vesicular-Arbuscular Mycorrhizal Fungus by Using Monoclonal Antibodies in an Enzyme-Linked Immunosorbent Assay

Spore morphology is currently used to identify species of vesicular-arbuscular mycorrhizal fungi. We report the first use of a highly specific immunological method for identification of a vesicular-arbuscular mycorrhizal fungus. Two monoclonal antibodies were produced against Glomus occultum. Monoclonal antibodies reacted strongly with both spores and hyphae in an indirect enzyme-linked immunosorbent assay. All other mycorrhizal (29 species) and nonmycorrhizal (5 species) fungi tested were nonreactive with the monoclonal antibodies. A single spore of G. occultum was detectable in the presence of high numbers of spores of other vesicular-arbuscular mycorrhizal fungi. Variation in the reaction of G. occultum isolates from West Virginia, Florida, and Colombia suggests that monoclonal antibodies may differentiate strains.     

西藏高原针茅草地土壤因子对丛枝菌根真菌物种多样性的影响

在对西藏高原北部针茅草地根围土壤中的丛枝菌根(AM)真菌种类分离鉴定基础上,研究了藏北针茅草地的土壤质地、pH、有机质和有效磷含量对AM真菌孢子密度、分离频度、相对多度、重要值、物种多样性指数和均匀度的影响.结果表明: 针茅草地根围土壤中共分离鉴定出AM真菌3属15种,其中,球囊霉属9种、无梗囊霉属6种、盾巨孢囊霉属1种.球囊霉属和无梗囊霉属为藏北针茅草地AM真菌的优势属;近明球囊霉和光壁无梗囊霉为藏北高寒草原针茅属植物根围AM真菌的优势种.不同质地土壤中AM真菌孢子密度、分离频度、相对多度和重要值均表现出球囊霉属＞无梗囊霉属＞盾巨孢囊霉属的趋势;土壤pH值对AM真菌种群组成无明显影响,球囊霉属和无梗囊霉属真菌分离频度、相对多度和重要值随土壤pH升高而增加,盾巨孢囊霉属则呈现相反趋势;不同土壤有机质含量范围内,AM真菌孢子密度等各项指标均呈球囊霉属＞无梗囊霉属＞盾巨孢囊霉属,而AM真菌属的分布没有明显规律;土壤有效磷含量对AM真菌种丰度和孢子密度影响较小.研究区域内AM真菌物种多样性指数和均匀度随着土壤有效磷含量升高而增加.     

钦州湾红树林丛枝菌根初步研究

海漆 (Excoecaria agallocha)、桐花树 (Aegiceras corniculatum)、秋茄 (Kandlia candel)、白骨壤 (Avi-cennia marina)是钦州湾分布广、生长量大的 4种红树植物 ,该项研究初步调查了丛枝菌根真菌在这四种植物根系的定居情况。在这四种植物的根际土壤均分离到丛枝菌根真菌孢子 ,其孢子密度以海漆根际的最高 ,其次为桐花树、秋茄、白骨壤。同时 ,海漆根系的侵染率和物种丰富度也最高 ,但在所取的白骨壤根样中没有观察到丛枝菌根的侵染。菌根侵染主要是以根内菌丝、胞内菌丝膨大扭曲、泡囊等形态出现。海水和土壤质地是影响菌根侵染率的主要因素。在所采土样中仅发现球囊霉属 (Glomus)、无梗囊霉属 (Acaulospora)丛枝菌根真菌 ,计有 1 7种和多个未确定种。     

Impact on Arbuscular Mycorrhiza Formation of Pseudomonas Strains Used as Inoculants for Biocontrol of Soil-Borne Fungal Plant Pathogens

The arbuscular mycorrhizal symbiosis, a key component of agroecosystems, was assayed as a rhizosphere biosensor for evaluation of the impact of certain antifungal Pseudomonas inoculants used to control soil-borne plant pathogens. The following three Pseudomonas strains were tested: wild-type strain F113, which produces the antifungal compound 2,4-diacetylphloroglucinol (DAPG); strain F113G22, a DAPG-negative mutant of F113; and strain F113(pCU203), a DAPG overproducer. Wild-type strain F113 and mutant strain F113G22 stimulated both mycelial development from Glomus mosseae spores germinating in soil and tomato root colonization. Strain F113(pCU203) did not adversely affect G. mosseae performance. Mycelial development, but not spore germination, is sensitive to 10 μM DAPG, a concentration that might be present in the rhizosphere. The results of scanning electron and confocal microscopy demonstrated that strain F113 and its derivatives adhered to G. mosseae spores independent of the ability to produce DAPG.     

Distribution of vesicular-arbuscular mycorrhizae in the plants and rhizosphere soils of the tropical plains,Tamil Nadu,India

We examined 737 plant species from 121 families of angiosperms and four species of pteridophytes for mycorrhizal association. Only 372 species showed infection. Mycorrhizal colonization was recorded in 49% of the total flora. The quantum of colonization ranged from 10% to 90%. In all, 35 species exhibited mycorrhizal colonization higher than 75%. Of the four pteridophytes, Isoetes coromandelina showed mycorrhizal colonization by Entrophospora schenckii and Glomus aggregation in its rhizosphere. For the first time in India, 102 of these species were reported to be mycorrhizal. Glomus fasciculatum (13.8%) and G. aggregation (11%) were prevalent in the rhizosphere soils. Acaulospora foveata (0.2%), A. longula (0.5%) and Glomus hoi (0.9%) were the least represented as mycorrhizal spores. A total of 40 vesicular-arbuscular mycorrhizal fungal species belonging to Acaulospora, Entrophospora, Gigaspora, Glomus, Sclerocystis and Scutellospora were isolated from the rhizosphere soils of different ecosystems.     

Phylogenetic position of an arbuscular mycorrhizal fungus,Acaulospora gerdemannii, and its synanamorphGlomus leptotichum, based upon 18S rRNA gene sequence

We examined the phylogenetic position of an arbuscular mycorrhizal fungus which produces two types of spore,Acaulospora gerdemannii andGlomus leptotichum, based upon the DNA sequence of the 18S rRNA gene. DNA was extracted separately from bothGlomus-like orAcaulospora-like spores and partial 5′-terminus segments of 18S rRNA gene were amplified by the PCR method. Several clones derived from each spore type were sequenced and compared. The sequences from both spore types agreed well, confirming that these morphologically different spores were formed by the same fungus. Nucleotide substitutions were found among several clones, suggesting polymorphism of the rRNA gene in glomalean fungi. Further phylogenetic analysis based upon the whole sequence of the 18S rRNA gene showed thatA. gerdemannii may be within the order Glomales but is far from the fungi that have been analyzed and probably should be in a new family.     

Seasonality of vesicular-arbuscular mycorrhizae in sedges in a semi-arid tropical grassland

Vesicular-arbuscular mycorrhizal (VAM) colonization and spore numbers in the rhizosphere of Cyperus iria L. and C. rotundus L., growing in a semi-arid tropical grassland, was studied during the 1993 and 1994 monsoons. In addition, climatic and chemical properties of the soils were determined in order to investigate their influence on mycorrhizal variables. VAM fungal association in the sedges was confirmed by plant- and root-trap culture techniques. The soil nutrients exhibited seasonal variations, but were highly variable between years. Intercellular hyphae and vesicles with occasional intraradical spores characterized mycorrhizal association in sedges. Dark septate fungi also colonized roots of sedges. Temporal variations in mycorrhizal colonization and spore numbers occurred, indicating seasonality. However, the patterns of mycorrhizal colonization and spore numbers were different during both the years. The VAM fungal structures observed were intercellular hyphae and vesicles. Changes in the proportion of root length with VAM structures, total colonization levels and spore numbers were related to climatic and edaphic factors. However, the intensity of influence of climatic and soil factors on VAM tended to vary with sedge species.     

四种蕨类植物根际土壤中VA菌根真菌孢子种群组成和季相变化

对４ 种移栽到温室中的蕨类植物根际土壤中的ＶＡ 菌根真菌孢子种群组成和季相变化进行了研究, 结果发现, ＶＡ菌根真菌孢子的产生具有明显的宿主依赖性和季相变化。在相同气候条件下, 不同植物根际土壤中的ＶＡ菌根真菌种群组成不同; 同种ＶＡ 菌根真菌在不同宿主植物根际土壤中, 孢子的丰富度有很大的差异。本文对影响ＶＡ菌根真菌孢子种群组成和季相变化的因素进行了讨论。     

Influence of edaphic and climatic factors on dynamics of root colonization and spore density of vesicular-arbuscular mycorrhizal fungi in Acacia farnesiana Willd. and A. planifrons W.et.A.

 A study was conducted to assess the dynamics of vesicular-arbuscular mycorrhizal (VAM) fungi associated with Acacia farnesiana and A. planifrons in moderately fertile alkaline soils. The intensity of root colonization by VAM fungi and the distribution of VAM fungal structures varied with host species over a period of time. The occurrence of vesicles with varied morphology in the mycorrhizal roots indicates infection by different VAM fungal species. This was further confirmed from the presence of spores belonging to different VAM fungal species in the rhizosphere soils. Root colonization and spore number ranged from 56% – 72% and 5 – 14 g^{–  1}soil in A. farnesiana and from 60% – 73% and 5 – 15 g^{–  1} soil in A. planifrons. Per cent root colonization and VAM spore number in the rhizosphere soil were inversely related to each other in both the Acacia species. However, patterns of the occurrence of VAM fungal structures were erratic. Spores of Acaulospora foveata, Gigaspora albida, Glomus fasciculatum, G. geosporum and Sclerocystis sinuosa were isolated from the rhizosphere of A. farnesiana whereas A. scrobiculata, G. pustulatum, G. fasciculatum, G. geosporum and G. microcarpum were isolated from that of A. planifrons. The response of VAM status to fluctuating edaphic factors varied with host species. In A. farnesiana though soil nitrogen (N) was positively correlated with root colonization, soil moisture, potassium and air temperature were negatively correlated to both root colonization and spore number. Per cent root colonization and spore number in A. planifrons were negatively related to each other. Further, in A. planifrons as the soil phosphorus and N were negatively correlated with the density of VAM fungal spores, the same edaphic factors along with soil moisture negatively influenced root colonization. Received: 16 May 1995 / Accepted: 7 February 1996     

Applying a solute transfer model to phytoextraction: Zinc acquisition by Thlaspi caerulescens

Phytoextraction is the removal of metals from contaminated soils into harvested plant tissues. The rate of phytoextraction is governed by both soil and plant characteristics. Most effort has focused on identifying appropriate plants for phytoextraction, but the benefits from this effort will be marginal unless the metals are in phytoavailable forms in the rhizosphere. The concentration of a metal in the rhizosphere can be estimated using solute transfer models that incorporate: the metal concentration in the bulk soil solution, the buffer power of the soil, diffusion coefficient for the metal, water movement, root size and morphology, and the rate of entry of metal into the roots. Here a solute transfer model is developed to predict the concentration of Zn in the rhizosphere solution ([Zn]_ext) of Thlaspi caerulescens, a hyperaccumulator species that could be exploited for Zn phytoextraction. The model predicts that Zn accumulation by T. caerulescens is sub-optimal when the Zn concentration in the bulk soil solution is <27 M. Such a high [Zn]_ext is rare in contaminated agricultural soils, but is possible in the metalliferous substrates where T. caerulescens is endemic. Sensitivity analyses indicate that Zn diffusion is more important than transpiration-driven mass flow for Zn delivery to the root, implying that management of soil physical and hydrological properties will improve phytoextraction. Sensitivity analyses also imply that strategies to enhance the Zn absorption power of the root will not necessarily be successful for enhancing phytoextraction per se. Thus, research into enhancing Zn availability and mobility in soil will be as important as understanding and manipulating Zn uptake by plants. In general, such models can be used to identify constraints to efficient phytoextraction (whether plant or soil) and to determine whether commercial phytoextraction is feasible.