Ultrastructure of rapidly frozen and freeze-substituted germ tubes of an arbuscular mycorrhizal fungus and localization of polyphosphate

In arbuscular mycorrhizas (AM), the supply of phosphorus from the fungi is one of the most important benefits to the host plant. Here we describe for the first time the ultrastructure and polyphosphate (poly P) distribution in rapidly frozen and freeze-substituted germ tubes of the AM fungus Gigaspora margarita. At the ultrastructural level, phosphorus distribution was analysed using energy-filtering transmission electron microscopy, and poly P was detected using an enzyme-affinity method. Semithin sections and live cells were also stained with 4',6-diamidino-2-phenylindole, which is not specific but fluoresces yellow when viewed under UV irradiation by binding with poly P. The cryotechnique method showed that extensive elongate ellipsoid vacuoles containing a uniform electron-opaque material occupied most of the cell volume. Combining the results of multiple methods revealed that poly P was localized in a dispersed form in vacuoles and in the outer fungal cell wall. These results show the significant potential of AM fungi for phosphorus storage based on its localization in the extensive complement of vacuoles in thick hyphae. The mechanism of translocation of poly P in tubular vacuoles, and the role of poly P in the cell wall, need to be elucidated.     

Depletion of soil mineral N by roots of Cucumis sativus L. colonized or not by arbuscular mycorrhizal fungi

Two experiments were conducted where Cucumis sativus were grown in uncompartmented pots either alone or in symbiosis with Glomus intraradices Schenck and Smith (Experiment 1) or Glomus sp. (Experiment 2) in order to investigate if root colonization by arbuscular mycorrhizal (AM) fungi has an effect on depletion of the soil mineral N pool. All pots were gradually supplied with 31 mg NH₄NO₃-N kg^-1 dry soil from 12–19 days after planting and an additional 50 mg (NH₄)₂SO₄-N kg^-1 dry soil (¹⁵N-labelled in Experiment 1) was supplied at 21 or 22 days after planting in Experiments 1 and 2, respectively. Dry weight of plant parts, total root length, mycorrhizal colonization rate and soil concentration of NH ₄ ⁺ and NO ₃ ^- were recorded at five sequential harvest events: 21, 24, 30, 35 and 42 days (Experiment 1) and 22, 25, 28, 31 and 35 days (Experiment 2) after planting. In Experiment 1, plants were also analysed for total content of N and ¹⁵N. The mycorrhizal colonization rate increased during time: from 25 to 40% in Experiment 1 and from 50 to 60% in Experiment 2. Plant dry matter accumulation was unaffected by mycorrhizal colonization, except in Experiment 1 where shoot dry weights were slightly increased and in Experiment 2 where root dry weights were slightly decreased compared to non-mycorrhizal control plants. The total root length was similar in the control and mycorrhizal treatments in Experiment 1, while it was decreased (20–30%) by mycorrhizal colonization in the last two harvest in Experiment 2. Mycorrhizal colonization affected the rate of depletion of soil mineral N in Experiment 1, where both NH ₄ ⁺ and NO ₃ ^- concentrations were markedly lower in the first two harvests, when plants were mycorrhizal. As the root length was similar in mycorrhizal and control treatments, this may indicate that the external AM hyphae contributed to the depletion of the soil mineral N pool. A similar pattern was observed in Experiment 2, although the effect was less pronounced. The ¹⁵N enrichment in mycorrhizal plants (Experiment 1) also indicated a faster NH ₄ ⁺ uptake than in the non-mycorrhizal controls in the first two harvests after application of the ¹⁵N-labelled N source. However, the external hyphae and roots seemed to have access to the same N sources as the ¹⁵N enrichment and total N content were similar in mycorrhizal and control plants at the end of the experiment. This revised version was published online in June 2006 with corrections to the Cover Date.     

丛枝菌根真菌对黄花蒿生长及药效成分的影响

通过盆栽接种试验,研究丛枝菌根（AM）真菌对药用植物黄花蒿的生长、营养吸收和药效成分的影响.结果表明：接种摩西球囊霉和地表球囊霉增加了黄花蒿对N、P、K的吸收,及叶片叶绿素含量、净光合速率、气孔导度、蒸腾速率、茎粗和地上生物量,尤其以接种摩西球囊霉的促进作用更强;接种摩西球囊霉后植株茎、小枝和叶中的青蒿素含量分别提高了32.8%、15.2%和19.6%,接种地表球囊霉后分别提高了26.5%、10.1%和14.9%;接种摩西球囊霉和地表球囊霉的黄花蒿地上部的挥发油收油率比未接种的分别提高45.0%和25.0%,而且挥发油成分发生了改变.     

水分胁迫下丛枝菌根真菌对红橘叶片活性氧代谢的影响

研究了水分胁迫下接种地表球囊霉（Glomus versiforme (Karsten) Berch）对红橘(Citrus tangerine Hort. ex Tanaka)叶片活性氧代谢的影响.结果表明：水分胁迫显著抑制了地表球囊霉对红橘根系的侵染,抑制率为33％.在正常供水和水分胁迫下,接种地表球囊霉处理的红橘叶片磷含量显著增加,与未接种处理相比,分别增加了45％和27%,丙二醛（MDA）和H₂O₂含量分别降低了25％、21％和16％、16％.正常供水和水分胁迫下接种地表球囊霉增强了叶片超氧化物岐化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)活性;提高了可溶性蛋白质、还原型抗坏血酸（ASC）和总抗坏血酸(TASC)含量.水分胁迫下接种处理显著降低了叶片超氧阴离子自由基（O₂^-_·）含量,与正常供水相比降低了31％.表明菌根化红橘植株的抗旱性增强.     

Medicago species affect the community composition of arbuscular mycorrhizal fungi associated with roots

Sequencing of the 5' end of the large ribosomal subunit (LSU rDNA) and quantitative polymerase chain reaction (qPCR) were combined to assess the impact of four annual Medicago species (Medicago laciniata, Medicago murex, Medicago polymorpha and Medicago truncatula) on the genetic diversity of arbuscular mycorrhizal (AM) fungi, and on the relative abundance of representative AM fungal genotypes, in a silty-thin clay soil (Mas d'Imbert, France). Two hundred and forty-six Glomeromycete LSU rDNA sequences from the four plant species and the bulk soil were analysed. The high bootstrap values of the phylogenetic tree obtained allowed the delineation of 12 operational taxonomic units (OTUs), all belonging to Glomus. Specific primers targeting Glomeromycetes and major OTUs were applied to quantify their abundance by qPCR. Glomeromycetes and targeted OTUs were significantly more abundant in the root tissues than in the bulk soil, and the frequencies of three of them differed significantly in the root tissues of the different plant species. These differences indicate that, despite the absence of strict host specificity in mycorrhizal symbiosis, there was a preferential association between some AM fungal and plant genotypes.     

AM真菌在有机农业发展中的机遇

在农田生态系统中,许多农作物均为丛枝菌根(AM)真菌的优良宿主植物,当AM真菌与这些宿主植物建立共生关系之后,AM真菌的存在有益于宿主植物的生长。然而,传统农业耕作模式中化学肥料和农药的施用、耕作制度的不断调整和非宿主植物的种植等都不利于AM真菌的建植。有机农业生态系统排除了化学肥料和农药的施用,减少了对AM真菌生长不利的因素,促进了土壤中AM真菌数量的增加和群落多样性的提高。同时,AM真菌可以通过多种方式改善土壤物理结构、提高农作物对干旱胁迫的耐受能力以及宿主植物对病虫害的抗性/耐性、抑制杂草生长、促进营养物质的吸收,进而提高植物的生长和改善产品的品质。基于此,围绕AM真菌在有机农业发展中的生态学功能展开论述,分析当前有机农业生态系统存在的问题,探讨利用AM真菌发展有机农业的可行性及其发展的机遇,以期促进AM真菌在有机农业发展中的应用。     

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.     

丛枝菌根真菌种间差异对柚苗营养生长及矿质含量的影响

在温室盆栽条件下研究了3种AM真菌Gigaspora margarita、Glomus mosseae和Glomus versiforme对长寿沙田柚(Citrus grandis cv. Changshou Shatian You)无菌苗营养生长及矿质含量的影响.结果表明,接种AM真菌的植株均有效地被感染;与对照相比,接种AM真菌能显著促进植株地上和地下部分生长,尤其促进了须根的生长,接种Glomus mosseae处理的主根长度比对照增加了22.7％,侧根数量增加了35.7％,须根数量和总长分别增加了160.8％和103.2％;接种AM真菌显著地提高了叶片的N、P、K、Ca、Mg、Zn、Cu和Mn含量,与对照相比,3种真菌处理的P含量分别增加了46.8％、88.7％和32.3％.3种AM真菌处理中,以接种Glomus mosseae处理营养生长最好,菌根依赖性最大,矿质元素N、P、K、Ca、Zn和Cu的含量最高,其效应顺序为Glomus mosseae＞Gigaspora margarita＞Glomus versiforme,可见,3种AM真菌对长寿沙田柚生长均有正效应,以Glomus mosseae最为显著,为长寿沙田柚适宜的优良菌种（株）.     

贵州烟草根围AM真菌多样性的初步研究

从贵州省内烟区不同土壤生态环境下采集烟草根际土样,湿筛离心法分离丛枝菌根（AM）真菌孢子,鉴定出烟草AM真菌4属20种,其中球囊霉属9种,无梗囊霉属7种,巨孢囊霉属3种,盾巨孢囊霉属1种。从土壤样品DNA中扩增AM真菌特异性片段并采用DGGE技术对AM真菌多样性进行分析。测序结果显示烟草根际土壤中菌根真菌主要菌群为球囊霉属,与湿筛离心法的鉴定结果一致。为进一步研究贵州地区AM真菌多样性以及开发应用提供了依据。     

Hyphal healing mechanism in the arbuscular mycorrhizal fungi Scutellospora reticulata and Glomus clarum differs in response to severe physical stress

The hyphal healing mechanism (HHM) has been shown to differ between Gigasporaceae and Glomeraceae. However, this process has not been considered under (severe) physical stress conditions. Scutellospora reticulata and Glomus clarum strains were cultured monoxenically. The impact of long distance separating cut extremities of hyphae and of multiple injuries within hyphae on the HHM was monitored. For long distances (>5000 microm) separating cut extremities, hyphae healing was observed in half the cases in S. reticulata and was absent in G. clarum. For multiple-injured hyphae, the HHM was always oriented towards the complete recovery of hyphae integrity in S. reticulata, while in G. clarum, the growing hyphal tips (GHTs) could indifferently reconnect cut sections, by-pass cut sections or develop into the environment. Hyphae behaviour under severe physical stress clearly differentiated S. reticulata from G. clarum, suggesting that both fungi have developed different strategies for colony growth to survive under adverse conditions.     

丛枝菌根真菌和根瘤菌对白三叶氮同化的影响

为揭示丛枝菌根真菌(AMF)和根瘤菌在白三叶氮(N)同化中的作用,该研究对白三叶进行单一或联合接种隐类球囊霉(Paraglomus occultum)和三叶草根瘤菌(Rhizobium trifolii),分析其对白三叶的生长、光合作用、叶片N和氨基酸含量以及N同化相关酶活性的影响。结果表明:(1)单一接种AMF或根瘤菌以及联合接种AMF和根瘤菌均显著增加了白三叶的株高、匍匐茎长度、叶片数、地上部生物量、总生物量、叶绿素b和总叶绿素含量、稳态光量子效率和叶片N含量,这种增强效应是联合接种>单一AMF>单一根瘤菌>未接种处理。(2)联合接种AMF和根瘤菌显著增加了白三叶叶片中丙氨酸、精氨酸、天冬酰胺、天冬氨酸、谷氨酰胺、谷氨酸和组氨酸的含量,显著提升了叶片N同化相关酶如硝酸还原酶、亚硝酸还原酶、谷氨酰胺合成酶、谷氨酸合成酶、谷氨酸脱氢酶、天冬酰胺合成酶和天冬氨酸转氨酶的活性,显著促进AMF对白三叶根系的侵染。综上认为,联合接种AMF和根瘤菌通过激活N同化相关酶活性有效促进N同化,产生更多氨基酸,进一步促进白三叶植株生长; 联合接种AMF和根瘤菌具有协同作用,有效促进了白三叶的N同化。     

Decline of arbuscular mycorrhizal fungi in northern hardwood forests exposed to chronic nitrogen additions

Arbuscular mycorrhizal (AM) fungi are important below-ground carbon (C) sinks that can be sensitive to increased nitrogen (N) availability. The abundance of AM fungi (AMF) was estimated in maple (Acer spp.) fine roots following more than a decade of experimental additions of N designed to simulate chronic atmospheric N deposition. Abundance of AMF was measured by staining and ocular estimation, as well as by analyzing for the AMF indicator fatty acid 16:1omega5c in phospholipid (biomass indicator) and neutral lipid (lipid storage indicator) fractions. Arbuscular mycorrhizal fungal biomass, storage structures and lipid storage declined in response to N addition measured by both methods. This pattern was found when AM response was characterized as colonization intensity, on an areal basis and in proportion to maple above-ground biomass. The phospholipid fraction of the fatty acid 16:1omega5c was positively correlated with total AMF colonization and the neutral lipid fraction with vesicle colonization. Decreased AMF abundance with simulated N deposition suggests reduced C allocation to these fungi or a direct soil N-mediated decline. The fatty acid (phospholipid and neutral lipid fractions) 16:1omega5c was found to be a good indicator for AMF active biomass and stored energy, respectively.     

塞北荒漠草原柠条锦鸡儿AM真菌的空间分布

为了探明塞北荒漠草原AM真菌生态分布规律,于2013年6月选取河北沽源县二牛点、内蒙古上都镇和青格勒图嘎查3个样地,按照0—10 cm、10—20 cm、20—30 cm、30—40 cm、40—50 cm 5个土层分别采集柠条锦鸡儿(Caragana korshinskii)根围土壤样品,研究了AM真菌空间分布特征及其与土壤因子的相关性。结果表明,柠条锦鸡儿根系能与AM真菌共生形成疆南星型丛枝菌根,AM真菌孢子密度和定殖率与样地和采样深度密切相关。二牛点孢子密度最大,3个样地孢子密度最大值均在0—10 cm土层,并随土层加深而减少;3个样地菌丝定殖率依次为上都镇青格勒图嘎查二牛点,峰值均在0—10 cm土层;泡囊定殖率青格勒图嘎查显著低于其他样地,但土层间无规律性变化;丛枝样地间定殖状况差异明显,变化趋势为青格勒图嘎查上都镇二牛点;AM真菌总定殖率和定殖强度最大值在上都镇。孢子密度与土壤有机C、全N、易提取球囊霉素和总球囊霉素极显著正相关,与p H值显著正相关,与速效P显著负相关;菌丝定殖率与土壤p H值、速效P、全N和酸性磷酸酶显著负相关;泡囊和丛枝定殖率与土壤碱解N和碱性磷酸酶具有极显著相关性;总球囊霉素和易提取球囊霉素与脲酶显著正相关,与碱解N、全N、碱性磷酸酶和酸性磷酸酶极显著正相关。主成分分析表明,酸性磷酸酶、总球囊霉素、全N、碱性磷酸酶、有机C是影响荒漠土壤营养状况的主要因子。总球囊霉素和易提取球囊霉素平均含量分别为3.19 mg/g和1.17 mg/g,占土壤有机C平均含量比为7.77%和3.83%,占土壤全N平均含量比为20.81%和9.57%。多元线性回归表明,总球囊霉素和易提取球囊霉素与土壤有机C和全N具有显著线性相关关系。研究球囊霉素与土壤有机C和N的比例关系可进一步明确AM真菌的生态功能,对荒漠土壤C库和N库研究具有重要意义。     

丛枝菌根真菌通过上调根系及自身水孔蛋白基因表达提高玉米抗旱性

在模拟干旱条件下, 研究了接种丛枝菌根(AM)真菌Glomus intraradices对玉米(Zea mays)根部13种质膜水孔蛋白基因表达的影响, 同时观测了AM真菌自身水孔蛋白基因的表达情况。结果表明, 干旱条件下, 除Zm PIP1;3、Zm PIP1;4、Zm PIP1;5和Zm PIP2;2之外的接种处理能显著提高根部其他8种质膜水孔蛋白基因的表达(Zm PIP2;7表达量未检测出), 并且AM真菌菌丝中水孔蛋白基因GintAQP1表达也显著增强。与此同时, 接种处理明显改善了植物水分状况, 提高了叶片水势。AM真菌增强宿主植物根部及自身的水孔蛋白基因的表达对于提高植物抗旱性具有潜在的重要贡献。