施肥对垂穗披碱草根系中丛枝菌根真菌的影响

采用传统染色与克隆测序的方法,研究了8年不同施肥(氮磷)梯度对垂穗披碱草根系中丛枝菌根(AM)侵染率和AM真菌群落的影响.结果表明: 随施肥浓度升高, 垂穗披碱草根系单位根长AM总侵染率从67.5%下降至7.3%,丛枝侵染率从5.2%降至0.1%.根系共检测出24个AM真菌分子种,但随着施肥浓度上升,AM真菌的平均物种丰富度从6种下降至2.6种.不同施肥处理对AM真菌群落结构有显著影响,土壤速效磷和根系氮含量与AM真菌群落呈极显著相关.氮磷有效性随施肥梯度逐渐上升,且与AM侵染率和AM真菌物种丰富度呈显著负相关.施高浓度氮磷肥对AM共生体有明显的抑制作用,导致AM真菌物种多样性丧失.     

丛枝菌根观察与侵染率测定方法的比较

菌根生长状况观察与侵染率测定是菌根学研究中一项重要的基础性工作。综述了丛枝菌根（AM）染色观察与侵染率测定方法研究概况,并对其进行比较和评价。认为采用醋酸墨水染色观察AM生长状况与采用根段侵染率加权法和放大交叉法测定AM真菌侵染率是目前较为科学、准确、易行的方法。根据不同需要也可选择其他适宜的方法,如要了解丛枝发育状况,可采用放大交叉法;如要了解泡囊和侵入点数量,可采用直接计数法,从而使其研究结果具有可比性。有必要建立基于分子生物学技术和脂肪酸定量分析技术测定一种或数种AM真菌侵染状况,这将有力推动AM真菌生理、生态功能研究的发展。     

典型农田土壤中丛枝菌根真菌-根际细菌互作及与氮磷利用的关系

在农业生态系统中,土壤微生物是土壤-作物系统养分循环的重要驱动力,其中丛枝菌根真菌(Arbuscularmycorrhizalfungi,AMF)能够促进作物对养分的吸收,适应逆境胁迫。【目的】进一步揭示AMF和根际细菌群落的跨界网络互作,挖掘与作物氮磷利用显著相关的关键微生物类群,揭示关键类群的生态网络特征。【方法】利用Illumina测序技术对3种典型农田旱地土壤(黑土、潮土和红壤)中AMF和根际细菌群落结构进行分析;构建互作网络并利用偏冗余分析、相关性分析探究了与氮磷利用相关的潜在关键类群。【结果】3种土壤中AMF与根际细菌均以正相互作用为主。不同土壤中AMF与根际细菌互作关系差异明显,在红壤中跨界互作最为密切,其中球囊霉属真菌(Glomus)与根际细菌中的放线菌(Actinobacteria)和变形菌(Proteobacteria)之间的交互作用最多。而在黑土中主要体现为根际细菌的界内互作。与氮磷利用率显著相关的关键微生物类群主要属于球囊霉属真菌、放线菌和α变形菌。【结论】典型旱地土壤中AMF与根际细菌的正相互关系对作物氮磷利用有潜在促进作用,关键类群在有机质和养分贫乏的红壤中可能起到更重要的作用。     

Copper sorption and accumulation by the extraradical mycelium of different Glomus spp. (arbuscular mycorrhizal fungi) isolated from the same polluted soil

The form and localisation of Cu accumulation in the extraradical mycelium (ERM) of three arbuscular mycorrhizal fungi (AMF), isolated from the same polluted soil contaminated with the Cu and Arsenate, was studied. There were differences in the capacity of the ERM of the three AMF to sorb and accumulate Cu. Glomus caledonium BEG133 had a significantly lower Cu-sorption capacity than Glomus mosseae BEG132 and Glomus claroideum BEG134 isolated from the polluted soil as well as an isolate of G. mosseae BEG25 from a non-polluted soil. This was directly related to the cation exchange capacity (CEC) of the ERM of these fungi. Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM) linked to an energy dispersive X-ray spectrometer (EDAX) gave more detailed information, showing that the ERM of AMF from the polluted soil was able to accumulate Cu in the mucilaginous outer hyphal wall zone, cell wall and inside the hyphal cytoplasm. The EDAX spectra showed that the accumulated Cu was mainly associated with Fe in the mucilaginous outer hyphal wall zone and in the cell wall. Cu was associated with traces of arsenate inside the cytoplasm of the ERM of Glomus mosseae BEG132 but this was not visible inside the ERM of Glomus caledonium BEG133 or Glomus claroideum BEG134. This work suggests that the ERM of AMF is able to sorb and accumulate Cu, but different tolerance mechanisms exist between the three AMF isolated from the same polluted soil providing further evidence for functional diversity within populations of AMF in soils.     

Effect of elevated atmospheric CO2 on mycorrhizal colonization, external mycorrhizal hyphal production and phosphorus inflow in Plantago lanceolata and Trifolium repens in association with the arbuscular mycorrhizal fungus Glomus mosseae

Plantago lanceolata and Trifolium repens were grown under ambient (400 μmol mol^–1) and elevated (650 μmol mol^–1) atmospheric CO₂ conditions. Plants were inoculated with the arbuscular mycorrhizal fungus Glomus mosseae and given a phosphorus supply in the form of bonemeal. Six sequential harvests were taken in order to determine whether the effect of elevated CO₂ on internal mycorrhizal colonization and external hyphal production was independent of the stimulatory effect of elevated CO₂ on plant growth. At a given time, elevated CO₂ increased the percentage of root length colonized (RLC), the total length of colonized root and the external mycorrhizal hyphal (EMH) density and decreased the ratio of EMH to total length of colonized root. When plant size was taken into account, the CO₂ effect on RLC and total length of colonized root was greatly reduced (and only apparent for early harvests in T. repens) and the effects on the EMH parameters disappeared. Root tissue P concentration was unchanged at elevated CO₂, but there was a decrease in shoot P at the later harvests. There was no direct effect of elevated CO₂ on P inflow for the earlier period (< 50 d) of the experiment. However, over the last period, there was a significant negative effect of elevated CO₂ on P inflow for both species, independent of plant size. It is concluded that elevated CO₂ had no direct effect on mycorrhizal colonization or external hyphal production, and that any observed effects on a time basis were due to faster growing plants at elevated CO₂. However, for older plants, elevated CO₂ had a direct negative effect on P inflow. This decrease in P inflow coincides with the observed decrease in shoot P concentration. This is discussed in terms of downregulation of photosynthesis often seen in elevated CO₂ grown plants, and the potential for mycorrhizas (via external hyphal turnover) to alleviate the phenomenon. The direction for future research is highlighted, especially in relation to carbon flow to and storage in the soil.     

Wetland dicots and monocots differ in colonization by arbuscular mycorrhizal fungi and dark septate endophytes

As an initial step towards evaluating whether mycorrhizas influence composition and diversity in calcareous fen plant communities, we surveyed root colonization by arbuscular mycorrhizal fungi (AMF) and dark septate endophytic fungi (DSE) in 67 plant species in three different fens in central New York State (USA). We found colonization by AMF and DSE in most plant species at all three sites, with the type and extent of colonization differing between monocots and dicots. On average, AMF colonization was higher in dicots (58±3%, mean±SE) than in monocots (13±4%) but DSE colonization followed the opposite trend (24±3% in monocots and 9±1% in dicots). In sedges and cattails, two monocot families that are often abundant in fens and other wetlands, AMF colonization was usually very low (<10%) in five species and completely absent in seven others. However, DSE colonization in these species was frequently observed. Responses of wetland plants to AMF and DSE are poorly understood, but in the fen communities surveyed, dicots appear to be in a better position to respond to AMF than many of these more abundant monocots (e.g., sedges and cattails). In contrast, these monocots may be more likely to respond to DSE. Future work directed towards understanding the response of these wetland plants to AMF and DSE should provide insight into the roles these fungal symbionts play in influencing diversity in fen plant communities.     

Seasonal changes of arbuscular mycorrhizal fungi as affected by tillage practices and fertilization : Hyphal density and mycorrhizal root colonization

The influence of tillage practices on native arbuscular mycorrhizal fungi (AMF) was studied in two, consecutive years in eastern Canada, in two 11 year-old long-term tillage-fertilizer experimental field soils, a sandy loam and a clay, growing corn in monoculture. The three tillage practices were: 1) conventional tillage (CT; fall plowing plus spring disking), reduced tillage (RT; spring disking) and no-till (NT). The corn crop received either inorganic (N and K) or organic (liquid dairy manure) fertilizers. Mycorrhizal hyphal density was estimated from soil samples obtained in early spring (before disking), at the 12–14 leaf stage, at silking, and at harvest. The percentage of corn root colonization by AMF at the 12–14 leaf stage, at silking and at harvest was also determined. The sandy loam was sampled over two consecutive seasons and the clay soil over one season.Densities of total and metabolically active soil hyphae, and mycorrhizal root colonization were significantly lower in CT soil than in RT and NT soil. Lowest soil hyphal densities were observed in early spring. The levels of intra- and extraradical fungal colonization always increased from spring to silking and decreased thereafter. Spring disking had only a small and transient negative effect on hyphal abundance in soil. Fertilization did not influence mycorrhizal colonization of corn or abundance of soil hyphae in the sandy loam soil, but in the clay soil metabolically active hyphae were more abundant with manure application than with mineral fertilization. In 1992, in both soils different tillage systems had same grain yield, however, in 1993, corn yield was higher in NT compared to CT system.     

Effect of glucose on the development ofGlomus fistulosum colonization and extraradical mycelium on maize roots

Various doses of glucose were added weekly to pots with maize growing in a mixture of soil and Perlite inoculated with the arbuscular mycorrhizal (AM) fungusGlomus fistulosum to define the effects of an additional carbon source on plant growth, mycorrhizae and other microbial features of the cultivation substratum. Higher doses of glucose (100 and 300 mg per pot) decreased plant growth and abundance of root hairs after 6 weeks of cultivation. Lower doses of glucose (10 and 30 mg per pot) had a positive effect on some characteristics of the development of the arbuscular fungus,e.g. root colonization, abundance of arbuscules, the length of extraradical mycelium associated with root surface, length of mycelium in the substratum as well as the length of both mycelia showing dehydrogenase activity as compared with variants not supplied with glucose solution. Glucose did not affect the number of spores of AM fungus. No effect of glucose was found on substratum respiration but glucose amendment increased microbial biomass and particularly the occurrence of saprophytic fungi. In a subsequent experiment focused on nonsymbiotic phase of fungal life cycle, the mycorrhizal root segments were incubated for 6 weeks in Petri dishes on membranes covered with a soil layer and supplied weekly with four glucose concentrations from 0.3 to 10 mg. Highest total length of hyphae associated with the root surface and the length of hyphae showing dehydrogenase activity was found when the lowest dose of 0.3 mg glucose was added to the soil weekly, whereas a 10 mg dose increased the length and activity of hyphae associated with a membrane. The possible mechanisms of the effects of additional labile carbon pool on the development of mycorrhizal fungus are discussed.     

Organic fertilizers alter the composition of pathogens and arbuscular mycorrhizal fungi in maize roots

Roots of agricultural crops, including maize, are hosts of different microorganisms, many beneficial, like plant growth and health‐promoting arbuscular mycorrhizal fungi (AMF), as well as pathogens including Pythium, Polymyxa and Microdochium. To improve crop nutrition and health, profound knowledge is required regarding how agricultural practices affect field populations of root‐associated microorganisms. Hence, the objective of this work was to evaluate the effect of crop genotype and organic fertilizers on the plant growth performance of maize and their root‐associated microorganisms. The experiment was conducted as a fully factorial greenhouse pot experiment with maize cultivars (two land races and two hybrids) and organic fertilizers (green manure, cow manure and compost) as the two main factors. Plants were harvested 8 weeks after sowing. In general, the different maize cultivars responded similarly to the applications of the organic fertilizers. Cow manure and compost increased plant growth, whereas green manure had limited effect on plant growth. Root colonization with AMF was reduced by green manure with rape. Infection with the root pathogens Pythium and Polymyxa was reduced by all organic fertilizers, whereas in contrast, infection with Microdochium increased with the majority of the organic fertilizers applied. In conclusion, both maize genotype and organic fertilizers affect the abundance of AMF and root pathogens in maize, which should be considered when developing management strategies of these root‐inhabiting microorganisms.     

The influence of Rhizobium and arbuscular mycorrhizal fungi on nitrogen and phosphorus accumulation by Vicia faba

BACKGROUND AND AIMS: The aim of this study was to investigate the effects of the interactions between the microbial symbionts, Rhizobium and arbuscular mycorrhizal fungi (AMF) on N and P accumulation by broad bean (Vicia faba) and how increased N and P content influence biomass production, leaf area and net photosynthetic rate. METHODS: A multi-factorial experiment consisting of four different legume-microbial symbiotic associations and two nitrogen treatments was used to investigate the influence of the different microbial symbiotic associations on P accumulation, total N accumulation, biomass, leaf area and net photosynthesis in broad bean grown under low P conditions. KEY RESULTS: AMF promoted biomass production and photosynthetic rates by increasing the ratio of P to N accumulation. An increase in P was consistently associated with an increase in N accumulation and N productivity, expressed in terms of biomass and leaf area. Photosynthetic N use efficiency, irrespective of the inorganic source of N (e.g. NO3- or N2), was enhanced by increased P supply due to AMF. The presence of Rhizobium resulted in a significant decline in AMF colonization levels irrespective of N supply. Without Rhizobium, AMF colonization levels were higher in low N treatments. Presence or absence of AMF did not have a significant effect on nodule mass but high N with or without AMF led to a significant decline in nodule biomass. Plants with the Rhizobium and AMF symbiotic associations had higher photosynthetic rates per unit leaf area. CONCLUSIONS: The results indicated that the synergistic or additive interactions among the components of the tripartite symbiotic association (Rhizobium-AMF-broad bean) increased plant productivity.     

Time-course study and partial characterization of a protein on hyphae of arbuscular mycorrhizal fungi during active colonization of roots

Material on the surface of hyphal walls of arbuscular mycorrhizal fungi (AMF) during active colonization of plant roots was detected by a monoclonal antibody. Pot-cultured isolates of Glomus, Acaulospora, Gigaspora, Scutellospora, and Entrophospora had immunofluorescent material (IM) on younger, thinner, intact hyphae, but IM was scant to absent on thicker, melanized or lysing hyphae. Colonization of corn (Zea mays L.), Sudangrass (Sorghum sudanense (Piper) Staph.) or red clover (Trifolium pratense L.) was examined during 5 months of plant growth by removing cores and performing an indirect immunoassay on roots with attached hyphae. Fresh spores of some Glomus spp. had IM on the outer layer of the spore wall. Abundant IM was seen on root hairs of plants colonized by some isolates, and some IM was detected on root surfaces of all plants examined even during early colonization. After cultures were dried, hyphae, roots and spores had little to no IM. Uninoculated control roots had very rare, small patches of IM. An immunoreactive protein was extracted from hyphae of Gigaspora and Glomus isolates by using 20mM citrate (pH 7.0) at 121°C for 90 min. Gel electrophoresis profiles indicated that all isolates tested had the same banding patterns. Lectin-binding of extracted protein is suggestive of a glycoprotein. The immunofluorescence assay can be used to examine root sections for active colonization by AMF, and the potential use of the protein to quantify AMF activity in soil is discussed.     

不同强度盐胁迫下AM真菌对羊草生长的影响

不同浓度NaCl盐处理下,AM真菌对羊草(Leymus chinensis)的侵染能力和对植物生长的影响,从植物形态和离子含量角度探讨了AM真菌提高羊草耐盐性的作用机理。结果表明,在高盐胁迫下,AM真菌显著降低了盐胁迫效应,提高了羊草生物量,菌根效应明显。菌根化羊草的根茎比显著增加,并且N、P浓度较高,Na~+和Cl~-离子浓度较低,表明AM真菌即促进羊草对营养元素的吸收,又减少了离子毒害。菌根化羊草的Ca~(2+)和K~+离子浓度,以及P/Na~+和K~+/Na~+比高于非菌根化羊草,表明AM真菌可通过调节渗透势以避免或减缓盐胁迫造成的生理缺水。随着盐胁迫的增加,菌根化羊草对磷的依赖性逐渐转换为对钾的依赖性。研究结果有助于揭示AM真菌提高植物耐盐能力的作用机理,并对应用菌根技术修复盐化草地具有理论指导意义。     

Extraradical mycelium network of arbuscular mycorrhizal fungi allows fast colonization of seedlings under in vitro conditions

Actively growing extraradical hyphae extending from mycorrhizal plants are an important source of inoculum in soils which has seldom been considered in vitro to inoculate young plantlets. Seedlings of Medicago truncatula were grown in vitro in the extraradical mycelium network extending from mycorrhizal plants. After 3, 6, 9, 12, and 15 days of contact with the mycelium, half of the seedlings were harvested and analyzed for root colonization. The other half was carefully transplanted in vitro on a suitable growth medium and mycelium growth and spore production were evaluated for 4 weeks. Seedlings were readily colonized after 3 days of contact with the mycelium. Starting from 6 days of contact, the newly colonized seedlings were able to reproduce the fungal life cycle, with the production of thousands of spores within 4 weeks. The fast mycorrhization process developed here opens the door to a broad range of in vitro studies for which either homogenous highly colonized seedlings or mass-produced in vitro inoculum is necessary. Liesbeth Voets and Ivan Enrique de la Providencia contributed equally to this work. MUCL is part of the Belgian Coordinated Collections of Micro-organisms (BCCM).     

设施栽培黄瓜根内AMF与DSE结构发育特征

本研究旨在观察和测定设施栽培黄瓜根系丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)与暗隔内生真菌(dark septate endophytic fungi,DSE)形态结构,明确其发育特征,为进一步探索AMF与DSE相互作用奠定基础。自山东莱阳、寿光和莱西等设施蔬菜主产区选择黄瓜Cucumis sativus样地,从不同连作年限、黄瓜生育期和土层深度分别采集黄瓜根系和根区土壤;观察根内AMF与DSE形态特征、测定AMF和DSE侵染数量、分析AMF或DSE侵染发育数量与黄瓜根结线虫Meloidogyne incognita病害的相关性。从黄瓜根系中可观察到典型的AMF泡囊、疆南星型(Arum,A)与重楼型(Paris,P)丛枝结构、DSE菌丝和微菌核。以黄瓜结果中期根系AMF和DSE侵染率最高,分别为57%和28%,苗期最低,分别为18%和8%;初花期的丛枝为P型,苗期和结果中期则为A型+P型。连作7年和7–10年的黄瓜根内丛枝为A型+P型,AMF和DSE的侵染率均分别显著高于连作10年的侵染率,连作10年的丛枝为A型。黄瓜根系以0–15cm土层中AMF侵染率最高(29%),丛枝为P型;以30cm的侵染率最低(12%),丛枝为A型;15–30cm土层的为A型+P型。AMF P型着生率、P/A比率和DSE侵染率分别与根结线虫病的为害程度具有相关性。研究结果还表明黄瓜根系AMF侵染率与DSE侵染率呈显著正相关关系。     

丛枝菌根真菌伴生细菌的研究进展

在丛枝菌根真菌（Arbuscular mycorrhizal fungi,AMF）的孢子、菌丝的表面或内部栖息着细菌,称之为AMF伴生细菌。AMF伴生细菌种类多样、分布广泛,生态位点包括孢子壁的表面或内部、细胞质、菌丝、孢子果等。其可能的生物学意义包括影响AMF孢子萌发、菌丝生长、菌根形成等过程。由于伴生细菌与AMF联系紧密,其对AMF和土壤微生物生态学具有重要的意义。国际上在该领域的研究已有30多年的历史,就其研究进展进行综述。