Community structure of arbuscular mycorrhizal fungi associated with Robinia pseudoacacia in uncontaminated and heavy metal contaminated soils

The significance of arbuscular mycorrhizal fungi (AMF) in soil remediation has been widely recognized because of their ability to promote plant growth and increase phytoremediation efficiency in heavy metal (HM) polluted soils by improving plant nutrient absorption and by influencing the fate of the metals in the plant and soil. However, the symbiotic functions of AMF in remediation of polluted soils depend on plant–fungus–soil combinations and are greatly influenced by environmental conditions. To better understand the adaptation of plants and the related mycorrhizae to extreme environmental conditions, AMF colonization, spore density and community structure were analyzed in roots or rhizosphere soils of Robinia pseudoacacia. Mycorrhization was compared between uncontaminated soil and heavy metal contaminated soil from a lead–zinc mining region of northwest China. Samples were analyzed by restriction fragment length polymorphism (RFLP) screening with AMF-specific primers (NS31 and AM1), and sequencing of rRNA small subunit (SSU). The phylogenetic analysis revealed 28 AMF group types, including six AMF families: Glomeraceae, Claroideoglomeraceae, Diversisporaceae, Acaulosporaceae, Pacisporaceae, and Gigasporaceae. Of all AMF group types, six (21%) were detected based on spore samples alone, four (14%) based on root samples alone, and five (18%) based on samples from root, soil and spore. Glo9 (Rhizophagus intraradices), Glo17 (Funneliformis mosseae) and Acau3 (Acaulospora sp.) were the three most abundant AMF group types in the current study. Soil Pb and Zn concentrations, pH, organic matter content, and phosphorus levels all showed significant correlations with the AMF species compositions in root and soil samples. Overall, the uncontaminated sites had higher species diversity than sites with heavy metal contamination. The study highlights the effects of different soil chemical parameters on AMF colonization, spore density and community structure in contaminated and uncontaminated sites. The tolerant AMF species isolated and identified from this study have potential for application in phytoremediation of heavy metal contaminated areas.     

Influence of Glomus etunicatum/Zea mays mycorrhiza on atrazine degradation, soil phosphatase and dehydrogenase activities, and soil microbial community structure

The effects of an arbuscular mycorrhizal (AM) fungus (Glomus etunicatum) on atrazine dissipation, soil phosphatase and dehydrogenase activities and soil microbial community structure were investigated. A compartmented side-arm (‘cross-pot’) system was used for plant cultivation. Maize was cultivated in the main root compartment and atrazine-contaminated soil was added to the side-arms and between them 650 or 37 μm nylon mesh was inserted which allowed mycorrhizal roots or extraradical mycelium to access atrazine in soil in the side-arms. Mycorrhizal roots and extraradical mycelium increased the degradation of atrazine in soil and modified the soil enzyme activities and total soil phospholipid fatty acids (PLFAs). Atrazine declined more and there was greater stimulation of phosphatase and dehydrogenase activities and total PLFAs in soil in the extraradical mycelium compartment than in the mycorrhizal root compartment when the atrazine addition rate to soil was 5.0 mg kg⁻¹. Mycelium had a more important influence than mycorrhizal roots on atrazine degradation. However, when the atrazine addition rate was 50.0 mg kg⁻¹, atrazine declined more in the mycorrhizal root compartment than in the extraradical mycelium compartment, perhaps due to inhibition of bacterial activity and higher toxicity to AM mycelium by atrazine at higher concentration. Soil PLFA profiles indicated that the AM fungus exerted a pronounced effect on soil microbial community structure.     

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。     

Hydrolytic enzyme activities in maize (Zea mays) and sorghum (Sorghum bicolor) roots inoculated with Gluconacetobacter diazotrophicus and Glomus intraradices

Two strains of Gluconacetobacter diazotrophicus (Pal 5, UAP5541) and the arbuscular mycorrhizal fungus Glomus intraradices increased both the shoot and root dry weight of sorghum 45 days after inoculation, whereas they had no effect on the shoot and root dry weight of maize. Co-inoculation (Gluconacetobacter diazotrophicus plus Glomus mosseae) did not increase the shoot and root dry weight of either plant. There was a synergistic effect of Gluconacetobacter diazotrophicus on root colonization of maize by Glomus intraradices, whereas an antagonistic interaction was observed in the sorghum root where the number of Gluconacetobacter diazotrophicus and the colonization by Glomus intraradices were reduced. Plant roots inoculated with Gluconacetobacter diazotrophicus and Glomus intraradices, either separately or together, significantly increased root endoglucanase, endopolymethylgalacturonase and endoxyloglucanase activities. The increase varied according to the plant. For example, in comparison with non-inoculated plants, there were higher endoglucanase (+328%), endopolymethylgalacturonase (+180%) and endoxyloglucanase (+125%) activities in 45-day old co-inoculated maize, but not in 45-day old sorghum. The possibility is discussed that hydrolytic enzyme activities were increased as a result of inoculation with Gluconacetobacter diazotrophicus, considering this to be one of the mechanisms by which these bacteria may increase root colonization by AM fungi.     

Interaction between the soil yeast Rhodotorula mucilaginosa and the arbuscular mycorrhizal fungi Glomus mosseae and Gigaspora rosea

The effect of the soil yeast, Rhodotorula mucilaginosa LBA, on Glomus mosseae (BEG n°12) and Gigaspora rosea (BEG n°9) was studied in vitro and in greenhouse trials. Hyphal length of G. mosseae and G. rosea spores increased significantly in the presence of R. mucilaginosa. Exudates from R. mucilaginosa stimulated hyphal growth of G. mosseae and G. rosea spores. Increase in hyphal length of G. mosseae coincided with an increase in R. mucilaginosa exudates. No stimulation of G. rosea hyphal growth was detected when 0.3 and 0.5 ml per petri dish of yeast exudates was applied. Percentage root length colonization by G. mosseae in soybean (Glycine max L. Merill) and by G. rosea in red clover (Trifolium pratense L. cv. Huia) was increased only when the soil yeast was inoculated before G. mosseae or G. rosea was introduced. Beneficial effects of R. mucilaginosa on arbuscular mycorrhizal (AM) colonization were found when the soil yeast was inoculated either as a thin agar slice or as a volume of 5 and 10 ml of an aqueous solution. R. mucilaginosa exudates (20 ml per pots) applied to soil increased significantly the percentage of AM colonization of soybean and red clover.     

Effect of soil bacteria on the ability of polycyclic aromatic hydrocarbons (PAHs) removal by Trametes versicolor and Irpex lacteus from contaminated soil

The effect of bacteria represented by indigenous soil microflora or a mixture of soil bacteria Pseudomonas aeruginosa and Rhodococcus erythropolis on fungal growth, extracellular enzyme production and polycyclic aromatic hydrocarbons (PAHs) biodegradation efficiency in soil of white-rot fungi Trametes versicolor and Irpex lacteus was investigated. Both fungi were able to colonize soil. The growth yields measured by ergosterol were about two-fold in I. lacteus after 10 weeks. Laccase was produced in T. versicolor cultures in the presence or absence of bacteria but live bacteria reduced the laccase levels in soil about 5 times. Manganese-dependent peroxidase (MnP) was not detected in T. versicolor cultures. The amounts of MnP and laccase in I. lacteus cultures were not affected by the presence of bacteria. T. versicolor was more efficient in PAH removal for all PAHs tested although its capacity to colonize soil was lower. The removal rates of PAHs by T. versicolor in sterile soil were 1.5-fold, 5.8-fold and 1.8-fold for 2-3-ring, 4-ring and 5-6-ring PAHs, compared to I. lacteus, respectively. I. lacteus showed a low efficiency of removal of pyrene, benzo[a]anthracene and benzo[k]fluoranthene, compared to T. versicolor, whereas chrysene and benzo[b]fluoranthene were degraded by neither fungus. The main effect of the presence of the indigenous microflora or R. erythropolis and P. aeruginosa was a significant decrease of degradation of total PAHs by both T. versicolor and I. lacteus. Weak fungal/bacterial synergistic effects were observed in the case of removal of acenapthylene, benzo[a]pyrene, dibenzo[a,h]anthracene and benzo[g,h,i]perylene by I. lacteus and acenapthylene by T. versicolor. However, the bacterial effects were different in the two fungi. PAH abiotic losses represented 15 and 21% of the total PAHs after 5 and 10 weeks, respectively; naphthalene and acenaphthene were removed from the soil due to volatilization.     

Soil inoculation with the biocontrol agent Clonostachys rosea and the mycorrhizal fungus Glomus intraradices results in mutual inhibition, plant growth promotion and alteration of soil microbial communities

Interactions between the biocontrol fungus Clonostachys rosea IK 726 and a tomato/Glomus intraradices BEG87 symbiosis were examined with and without wheat bran, which served as a food base for C. rosea. In soil without wheat bran amendment, inoculation with C. rosea increased plant growth and altered shoot nutrient content resulting in an increase and decrease in P and N content, respectively. Inoculation with G. intraradices had no effect on plant growth, but increased the shoot P content. Dual inoculation with G. intraradices and C. rosea followed the pattern of C. rosea in terms of plant growth and nutrient content. Wheat bran amendment resulted in marked plant growth depressions, which were counteracted by both inoculants and dual inoculation increased plant growth synergistically. Amendment with wheat bran increased the population density of C. rosea and reduced mycorrhizal fungus colonisation of roots. The inoculants were mutually inhibitory, which was shown by a reduction in root colonisation with G. intraradices in treatments with C. rosea and a reduction in colony-forming units (cfu) of C. rosea in treatments with G. intraradices, irrespective of wheat bran amendment. Moreover, both inoculants markedly influenced soil microbial communities examined with biomarker fatty acids. Inoculation with G. intraradices increased most groups of microorganisms irrespective of wheat bran amendment, whereas the influence of C. rosea on other soil microorganisms was affected by wheat bran amendment. Overall, inoculation with C. rosea increased and decreased most groups of microorganisms without and with wheat bran amendment, respectively. In conclusion, despite mutual inhibition between the two inoculants this interaction did not impair their observed plant growth promotion. Both inoculants also markedly influenced other soil microorganisms, which should be further studied in relation to their plant growth-promoting features.     

土壤重金属污染的植物修复与金属超富集植物及其遗传工程研究

本文综述了土壤重金属污染的植物修复、金属超富集植物及其遗传工程的最新研究进展及存在问题,并提出加紧筛选和发现野生高生物量的金属超富集植物,在现有高生物量作物种质资源中筛选金属超富集作物,应用遗传工程技术把野生植物的超富集基因转移到现有高生物量植物(作物)中,寻找综合、可持续的植物修复手段等对策。     

丛枝菌根真菌对玉米和续断菊间作镉吸收和累积的影响

通过盆栽试验,利用分室隔网培养方法,模拟研究接种丛枝菌根真菌(AMF)对玉米-续断菊(Sonchus asper L.Hill)间作体系Cd吸收和累积的影响。结果表明:1无论Cd添加到A室还是B室,玉米的侵染率都要比续断菊高出6.3%~38.35%。接种AMF之后,都不同程度地提高了玉米和续断菊的生物量,但对玉米和续断菊吸收Cd的影响有所不同。2Cd添加到A或者B室,AMF均会促进该侧植物对Cd的吸收,降低另一侧植物对Cd的吸收。3接种AMF后,续断菊对Cd的转运系数降低,玉米对Cd的转运系数有的升高有的降低。总之,AMF改变了间作条件下玉米和续断菊对Cd的吸收,菌丝在两者之间可能起着非常重要的作用。     

Barley yield and soil microbial and enzyme activities as affected by contamination of two soils with lead, zinc or copper

The effects of increasing rates of Pb, Zn and Cu on extractable heavy metal levels, barley yields, basal respiration and the activities of catalase, urease, invertase and acid phosphatase were investigated in two soils in a 2-year greenhouse experiment. In the first year, barley yields were decreased by increasing additions of Pb, Zn and Cu. In the second year, increased yields were recorded at lower rates of addition of all three metals in both the chestnut and chernozem soils. Yield depressions were most marked for added Cu and least marked for Pb but, in contrast, accumulation of heavy metals in grain, in excess of recommended limits, was most pronounced for Pb and did not occur for Cu. Increasing rates of all three metals caused a decrease in basal respiration; the degree of inhibition was generally greater in the second than in the first year. After 1 year of incubation, increasing rates of addition of metals reduced all tested enzyme activities. However, after 2 years, the pattern of response was more complex, with increases in enzyme activities being noted at lower rates of addition of all three metals. In general, invertase and urease activities were more markedly inhibited by heavy metal contamination than those of catalase and phosphatase. Ammonium acetate-extractable heavy metal concentrations in soils were less after 2 years than 1 years reaction time due to their transformation into less labile forms. Significant negative correlations between grain yield, basal respiration and enzyme activities were observed in both years.     

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.     

The mycorrhizal fungus Glomus intraradices and rock phosphate amendment influence plant growth and microbial activity in the rhizosphere of Acacia holosericea

Plants inoculated with arbuscular mycorrhizal (AM) fungi utilize more soluble phosphorus from soil mineral phosphate than non-inoculated plants. However, there is no information on the response of soil microflora to mineral phosphate weathering by AM fungi and, in particular, on the catabolic diversity of soil microbial communities.The AM fungus, Glomus intraradices was examined for (i) its effect on the growth of Acacia holosericea, (ii) plant-available phosphate and (iii) soil microbial activity with and without added rock phosphate.After 4-months culture, AM fungal inoculation significantly increased the plant biomasses (by 1.78× and 2.23× for shoot and root biomasses, respectively), while mineral phosphate amendment had no effect in a sterilized soil. After 12-months culture, the biomasses of A. holosericea plants growing in a non-sterilized soil amended with mineral phosphate were significantly higher than those recorded in the control treatment (by 2.5× and 5× for shoot and root biomasses, respectively). The fungal inoculation also significantly stimulated plant growth, which was significantly higher than that measured in the mineral phosphate treatment. When G. intraradices and mineral phosphate were added together to the soil, shoot growth were significantly stimulated over the single treatments (inoculation or amendment) (1.45×). The P leaf mineral content was also higher in the G. intraradices+mineral phosphate treatment than in G. intraradices or rock phosphate amendment. Moreover, the number of fluorescent pseudomonads has been significantly increased when G. intraradices and/or mineral phosphate were added to the soil. By using a specific type of multivariate analysis (co-inertia analysis), it has been shown that plant growth was positively correlated to the metabolization of ketoglutaric acid, and negatively linked to the metabolisation of phenylalanine and other substrates, which shows that microbial activity is also affected.G. intraradices inoculation is highly beneficial to the growth of A. holosericea plants in controlled conditions. This AM symbiosis optimises the P solubilization from the mineral phosphate and affects microbial activity in the hyphosphere of A. holosericea plants.     

Inoculation with arbuscular mycorrhizal fungus Acaulospora mellea decreases Cu phytoextraction by maize from Cu-contaminated soil

A pot culture experiment was carried out to study the growth of and Cu uptake by maize (Zea mays) inoculated with or without arbuscular mycorrhizal (AM) fungus Acaulospora mellea in sterilized soil with different Cu amounts added (0, 100, 200, 400, 800 mg kg⁻¹). Root colonization rates were significantly lower with the addition of 400 and 800 mg kg⁻¹ Cu. AM inoculation increased shoot dry weights at 200 and 400 mg kg⁻¹ Cu added but showed no effects at other levels, while increased root dry weights at all Cu addition levels except 800 mg kg⁻¹. Compared with the nonmycorrhizal plants, shoot Cu concentrations in mycorrhizal plants were higher when no Cu was added but lower at other levels, while root Cu concentrations were lower at 400 and 800 mg kg⁻¹ Cu added but not affected at other levels. Thus, shoot Cu uptake in mycorrhizal plants increased with no Cu added but decreased at other levels, while mycorrhizal effects on root Cu uptake varied. Compared with nonmycorrhizal controls, Cu uptake efficiency and phytoextraction efficiency in mycorrhizal plants were higher when no Cu was added but lower at other levels, and Cu translocation efficiency was lower at all Cu addition levels. AM inoculation improved shoot and root P nutrition at all Cu addition levels. Soil pH was higher in mycorrhizal treatment than in the control when 200 mg kg⁻¹ or more Cu was added. These results indicate that A. mellea ZZ may be not suitable for Cu phytoextraction by maize, but shows a potential role in phytostabilization of soil moderately polluted by Cu.     

Accumulation of specific flavonoids in soybean (Glycine max (L.) Merr.) as a function of the early tripartite symbiosis with arbuscular mycorrhizal fungi and Bradyrhizobium japonicum (Kirchner) Jordan

This study is the first report assessing the effect of soil inoculation on the signalling interaction of Bradyrhizobium japonicum, arbuscular mycorrhizal fungi (AMF) and soybean plants throughout the early stages of colonisation that lead to the tripartite symbiosis. In a study using soil disturbance to produce contrasting indigenous AMF treatments, the flavonoids daidzein, genistein and coumestrol were identified as possible signals for regulating the establishment of the tripartite symbiosis. However, it was unclear whether soil disturbance induced changes in flavonoid root accumulation other than through changing the potential for AMF colonization. In this study, soil treatments comprising all possible combinations of AMF and B. japonicum were established to test whether (1) modifications in root flavonoid accumulation depend on the potential for AMF colonization, and (2) synthesis and accumulation of flavonoids in the roots change over time as a function of the early plant-microbial interactions that lead to the tripartite symbiosis. The study was comprised of two phases. First, maize was grown over 3-week periods to promote the development of the AM fungus Glomus clarum. Second, the interaction between soybean, G. clarum and B. japonicum was evaluated at 6, 10, 14 and 40 days after plant emergence. Root colonization by G. clarum had a positive effect on nodulation 14 days after emergence, producing, 30% more nodules which were 40% heavier than those on roots solely inoculated with B. japonicum. The tripartite symbiosis resulted in 23% more N₂ being fixed than did the simpler symbiosis between soybean and B. japonicum. The presence of both symbionts changed accumulation of flavonoids in roots. Daidzein and coumestrol increased with plant growth. However, development of the tripartite symbiosis caused a decrease in coumestrol; accumulation of daidzein, the most abundant flavonoid, was reduced in the presence of AMF.     

Nickel-tolerant Brevibacillus brevis and arbuscular mycorrhizal fungus can reduce metal acquisition and nickel toxicity effects in plant growing in nickel supplemented soil

The growth of clover (Trifolium repens ) and its uptake of N, P and Ni were studied following inoculation of soil with Rhizobium trifolii, and combinations of two Ni-adapted indigenous bacterial isolates (one of them was Brevibacillus brevis) and an arbuscular mycorrhizal (AM) fungus (Glomus mosseae). Plant growth was measured in a pot experiment containing soil spiked with 30 (Ni I), 90 (Ni II) or 270 (Ni III) mg kg⁻¹ Ni-sulphate (corresponding to 11.7, 27.6 and 65.8 mg kg⁻¹ available Ni on a dry soil basis). Single inoculation with the most Ni-tolerant bacterial isolate (Brevibacillus brevis) was particularly effective in increasing shoot and root biomass at the three levels of Ni contamination in comparison with the other indigenous bacterial inoculated or control plants. Single colonisation of G. mosseae enhanced by 3 fold (Ni I), by 2.4 fold (Ni II) and by 2.2 fold (Ni III) T. repens dry weight and P-content of the shoots increased by 9.8 fold (Ni I), by 9.9 fold (Ni II) and by 5.1 fold (Ni III) concomitantly with a reduction in Ni concentration in the shoot compared with non-treated plants. Coinoculation of G. mosseae and the Ni-tolerant bacterial strain (B. brevis) achieved the highest plant dry biomass (shoot and root) and N and P content and the lowest Ni shoot concentration. Dual inoculation with the most Ni-tolerant autochthonous microorganisms (B. brevis and G. mosseae) increased shoot and root plant biomass and subtantially reduced the specific absorption rate (defined as the amount of metal absorbed per unit of root biomass) for nickel in comparison with plants grown in soil inoculated only with G. mosseae. B. brevis increased nodule number that was highly depressed in Ni I added soil or supressed in Ni II and Ni III supplemented soil. These results suggest that selected bacterial inoculation improved the mycorrhizal benefit in nutrients uptake and in decreasing Ni toxicity. Inoculation of adapted beneficial microorganisms (as autochthonous B. brevis and G. mosseae) may be used as a tool to enhance plant performance in soil contaminated with Ni.     

不同土壤类型下AM 真菌分布多样性及与土壤因子的关系

以禾本科植物群落为研究对象, 研究了宁夏六盘山林地、银川农耕地、暖泉农耕地、固原农耕地、盐池沙地、灵武沙地6 个采样地点5 种土壤类型(黑垆土、灌淤土、黄绵土、灰钙土、风沙土)下AM 真菌物种多样性及其与土壤因子的关系。结果表明: 5 种土壤类型采样点的植被根际土壤中共鉴定出5 属48 种AM真菌, 其中, 无梗囊霉属(Acaulospora)1 种, 巨孢囊霉属(Gigaspora)3 种, 球囊霉属(Glomus)37 种, 类球囊霉属(Paraglomus)1 种, 盾巨孢囊霉属(Scutellospora)6 种, 各采样点土壤均以球囊霉属为优势属。地球囊霉(G.geosporum)和木薯球囊霉(G. manihotis)是6 个采样地点中的优势种。不同土壤类型各采样点AM 真菌各属的频度存在明显差异, 球囊霉属在各点均有出现, 频度值最高。具有较高植被多样性的暖泉样点, AM 真菌的种属数量较多。土壤环境因子对AM 真菌孢子密度的影响因所处土壤、植被类型不同而异。pH、全盐、速效钾、速效磷等土壤肥力因子, 在PCA 轴上能最大程度地解释AM 真菌孢子密度与土壤环境因子之间相互关系的大部分信息。宁夏不同土壤类型区域中AM 真菌种类及分布一定程度上与该采样点的植被类型、植物多样性和土壤肥力特征相对应。     

The bacterial community of tomato rhizosphere is modified by inoculation with arbuscular mycorrhizal fungi but unaffected by soil enrichment with mycorrhizal root exudates or inoculation with Phytophthora nicotianae

Arbuscular mycorrhizal (AM) fungi have been shown to induce the biocontrol of soilborne diseases, to change the composition of root exudates and to modify the bacterial community structure of the rhizosphere, leading to the formation of the mycorrhizosphere. Tomato plants were grown in a compartmentalized soil system and were either submitted to direct mycorrhizal colonization or to enrichment of the soil with exudates collected from mycorrhizal tomato plants, with the corresponding negative controls. Three weeks after planting, the plants were inoculated or not with the soilborne pathogen Phytophthora nicotianae growing through a membrane from an adjacent infected compartment. At harvest, a PCR-Denaturing gradient gel electrophoresis analysis of 16S rRNA gene fragments amplified from the total DNA extracted from each plant rhizosphere was performed. Root colonization with the AM fungi Glomus intraradices or Glomus mosseae induced significant changes in the bacterial community structure of tomato rhizosphere, compared to non-mycorrhizal plants, while enrichment with root exudates collected from mycorrhizal or non-mycorrhizal plants had no effect. Our results support that the effect of AM fungi on rhizosphere bacteria would not be mediated by compounds present in root exudates of mycorrhizal plants but rather by physical or chemical factors associated with the mycelium, volatiles and/or root surface bound substrates. Moreover, infection of mycorrhizal or non-mycorrhizal plants with P. nicotianae did not significantly affect the bacterial community structure suggesting that rhizosphere bacteria would be less sensitive to the pathogen invasion than to mycorrhizal colonization. Of 96 unique sequences detected in the tomato rhizosphere, eight were specific to mycorrhizal fungi, including two Pseudomonas, a Bacillus simplex, an Herbaspirilium and an Acidobacterium. One Verrucomicrobium was common to rhizospheres of mycorrhizal plants and of plants watered with mycorrhizal root exudates.     

重金属污染农田安全利用:进展与展望

我国耕地土壤污染面积广,污染情况复杂,农产品重金属超标问题已经关系到国计民生。常用的物理化学修复方法成本高,不适用于大面积的中低污染农田。植物提取修复方法成本低,环境友好,但修复时间长,推广困难。总的来讲,基于重金属移除的诸技术在解决农田重金属污染方面还没有太大优势。相较而言,农田安全利用在不移除或缓慢移除土壤重金属的条件下,以生产安全农产品为目标,具有更加坚实的现实意义和推广价值。种植低吸收农作物是安全利用的重要措施,基因工程手段在低吸收农作物品种筛选中具有巨大的潜力,但其可能带来的生态环境风险使得这些通过基因工程得到的低吸收作物的田间种植面临着巨大挑战。土壤添加剂可以改变土壤重金属形态,降低重金属的生物有效性,但会对土壤质量产生影响。微生物尤其是土著微生物的利用越发受到关注,改变微生物的生存环境与基因工程手段能够强化微生物的钝化效果。施肥、水分管理、间作等农艺措施也能改变土壤重金属的形态,抑制作物对重金属的吸收。未来以加强推广为目的,多种技术手段的联合应用是重金属污染农田安全利用的重要发展方向,其中以生物技术为核心的利用模式具有十分重要的意义。     

The role of arbuscular mycorrhizal colonization in the carbon and nutrient economy of the tripartite symbiosis with nodulated Phaseolus vulgaris

In the tripartite symbiosis between nodulated legume roots and arbuscular mycorrhizal (AM) fungi, symbiont sink strength may depend upon developmental stage and the nutrient benefits to the host plant. The cost-benefits of the tripartite symbiosis were investigated in terms of C-economy and nutrition. Nodulated Phaseolus vulgaris seedlings, with and without AM, were hydroponically grown under high (2 mM) and low (1 μM) P conditions in an N-free Long Ashton nutrient solution. Plants were sequentially harvested at 17, 24 and 31 days after emergence. At each harvest, measurements for biomass, N₂-fixation, photosynthesis, root respiration, calculated C and nutritional economy were taken. Nodular growth was suppressed by the early development of AM colonization. This coincided with higher photosynthetic and respiratory rates in AM plants. These effects were most pronounced under low P when AM colonization peaked. Once AM levels reached the plateau phase, the efficiency of P nutrition increased. This was followed by improved nodular and host growth and enhanced N₂-fixation. This indicates that the AM was the dominant symbiont for host C in the tripartite symbiosis, due to its rapid development and subsequent role in supplying P more effectively to both host and nodules.     

Specific flavonoids as interconnecting signals in the tripartite symbiosis formed by arbuscular mycorrhizal fungi, Bradyrhizobium japonicum (Kirchner) Jordan and soybean (Glycine max (L.) Merr.)

Many legume plants benefit from the tripartite symbiosis of arbuscular mycorrhizal fungi (AMF) and rhizobia. Beneficial effects for the plant have been assumed to rely on increased P supply through the mycorrhizas. Recently, we demonstrated that P does not regulate the establishment of the tripartite symbiosis. Flavonoids appear to play a role as early signals for both rhizobia and AMF. Four soybean lines known to express different concentrations of the isoflavones genistein, daidzein and glycitein in the seed were used to test three hypotheses: (i) The establishment of the tripartite symbiosis is not dependent of a nutrient mediated effect; (ii) There is a positive correlation between seed isoflavone concentrations of different soybean lines and the progress of the tripartite symbiosis; (iii) Specific flavonoids control the establishment of the tripartite symbiosis in that a change in flavonoid root accumulation resulting from the development of one microbial partner can stimulate colonization of soybean roots by the other. Disturbed versus undisturbed soil treatments were produced to vary the potential for indigenous AMF colonization of soybean. In contrast, the potential for Bradyrhizobium was kept identical in both soil disturbance treatments. The uptake of P and Zn and the concentration of flavonoids in mycorrhizal soybean roots at 10 d after emergence were analysed either separately of Bradyrhizobium or in context of the tripartite symbiosis. Zinc nutrition did not differ between AMF treatments which supports the first hypothesis. The concentration of daidzein was at least four times greater in the root than in the seed reaching 3958±249 μg g⁻¹ dry across soybean lines. Coumestrol, which was absent in the seed, was synthesized to reach 2154±64 μg g⁻¹ dry. Conversely, the concentration of genistein was approximately three times smaller in the root that in the seed (301±15 μg g⁻¹ dry), while glycitein and formononetin were never detected. The establishment of the tripartite symbiosis was identical across soybean lines which does not support the second hypothesis. Concentrations of flavonoids were significantly greater in roots under disturbed soil, for which both symbioses were not as developed as in plants from undisturbed soil. This clearly supports the third hypothesis. This research provides the first data linking the function of different flavonoids to the establishment of the tripartite symbiosis, and suggests that these compounds are produced and released into the rhizosphere as a function of the colonization process.