长期定位施肥土壤中的丛枝菌根真菌

从连续26年长期定位施肥土壤中的玉米和小麦根围采集土样,从中鉴定出丛枝茵根(Arbuscular Myc- orrguza,AM)真菌5属19种,包括Glomus属11种:G．caledonium,G．constrictum,G．eburneum,G．ebunicatum, G．hyderabadensis,G．mossese,G．sinuosum,G．tortuosum,G．versiforme,G．sp1,G．sp2;Acaulospora属2种: A．excacota,A．sp;Gigaspora属2种:Gi．margarita,Gi．sp;Scutellospora属3种:S．cerradensis,S．coralloidea, S．pellucida;Archaespora属1种:Ar．leptoticha。     

菌根真菌在红壤中对柑桔吸收磷肥的研究

接种一种球囊霉 Glomus sp.编号 CC-1于红壤中,使酸桔和枳实生苗的根系感染形成泡囊丛枝内生菌根。应用放射性同位素~(32)P 标记的可溶性磷肥和难溶性磷肥来研究菌根真菌对柑桔吸收磷肥的作用。放射性宏观自显影和显微自显影表明菌根真菌加强了柑桔对两类磷肥的吸收。放射性测量和化学分析证明有菌根柑桔实生苗的地上部含磷量以及从红壤中和从肥料中吸收的磷量比无菌根的增多,有菌根实生苗的生长反应也较好。     

外源养分和激素对AM真菌侵染和产孢的影响

在温室30%强度Hoagland营养液半水培条件下,对寄主植物玉米Zea mays接种丛枝菌根(Arbuscular myeorrhizae,AM)真菌Glomus mosseae后施加不同浓度的葡萄糖、蔗糖、脯氨酸(Pro)、吲哚-3-乙酸(IAA)、赤霉素(GA3)和6-苄基腺嘌呤(6-BA)。结果表明:除低浓度GA3处理外,其他处理尤其是高浓度处理抑制了玉米植株的生长。中浓度蔗糖、脯氨酸(Pro)和吲哚-3-乙酸(IAA)处理显著提高玉米菌根侵染率,其中以中浓度蔗糖的处理最高,达92.3%。葡萄糖、赤霉素(GA3)和6-苄基腺嘌呤(6-BA)处理则降低菌根侵染率。施加不同有机养分和植物生长物质并未提高AM真菌产孢数量,相反,却减少了孢子的形成。高浓度氨基乙酸(Gly)和低浓度6-BA处理的产孢量分别为50和48个/管,高于其他处理,但低于对照(78个/管);可溶性糖处理的产孢数量最少,平均仅18个/管。本试验条件下菌根侵染率与产孢数量不具有相关性,而产孢数与根外菌丝量具相关性。认为通过外界不同的化学调控,对植物生长、菌根侵染、根外菌丝量和孢子发育产生了一定的影响,具有一定的调节作用。     

我国北方农田土壤中AM真菌的多样性

AM真菌是农业生态系中一类重要的土壤微生物,它在农田土壤中的发生和分布受多种环境因素的影响。为深入了解我国北方农田土壤中AM真菌的多样性规律,于2000年在河北、山东的农田土壤中采集有代表性的土样127个。通过进一步扩繁、纯化,从中分离出AM真菌5属22种,鉴定了20个种,包括一个国内新记录种沾屑球囊霉(Glomus spurcum)。分析AM真菌的多样性特点及其影响因素发现,农田土壤中以球囊霉属(Glomus)的频度最高,其次为无梗囊霉属(Acaulospora);优势种类为幼套球囊霉(Glomus etunicatum)和摩西球囊霉(Glomus mosseae).土壤速效磷含量、pH状况主要对孢子密度产生影响,对种群分布影响不大。宿主植物类型对AM真菌的侵染状况和多样性影响较大;比较玉米(Zea mays)、甘薯(Ipomoea batatas)根区AM真菌的种群组成后发现两者有所不同,但优势种一致.     

城市生态系统中AM真菌侵染与群落结构特征

丛枝菌根(AM)真菌作为土壤微生物的重要成员之一,对城市生态系统可持续发展具有重要意义.本文系统总结了城市生态系统中AM真菌着生状况和群落结构特点,探讨了城市生态因子,如人类行为、植被重建与维护、城市土壤状况等对AM真菌着生状况和群落结构的影响,认为今后应加强城市生态系统中AM真菌群落结构与功能的研究,如关键城市生态因子(如水资源匮乏、热岛效应等)改变AM真菌群落结构的效应与机制.     

盐胁迫下接种AM真菌对玉米耐盐性的影响

以玉米品种陕单16号幼苗为材料,用盆栽法研究了不同含盐量(0、0.5、1.0、1.5和2.0 NaCl g/kg)土壤接种AM真菌(Glomus mosseae)对玉米幼苗生物量、盐害级数,以及叶片中电解质透出率、丙二醛、O·2-、H2O2含量和保护酶活性的影响.结果表明:在盐胁迫下,接种AM真菌增加了玉米植株生物量,降低了玉米的盐害级数;菌根植株叶片中过氧化氢酶的活性高于非菌根植株,而过氧化物酶、抗坏血酸氧化酶和多酚氧化酶的活性则为非菌根植株高于菌根植株;超氧化物歧化酶的活性在NaCl浓度为0、0.5和1.0 g/kg时为非菌根植株高于菌根植株,而在NaCl浓度为1.5和2.0 g/kg时则为菌根植株高于非菌根植株;菌根植株叶片中电解质透出率、丙二醛、O·2-和H2O2的含量低于非菌根植株.可见,AM真菌的侵染提高了玉米的耐盐性,缓解了由盐胁迫引起的过氧化胁迫对玉米植株的伤害,但这一缓解作用并不只是通过提高保护酶活性来实现的,可能还存在一些非酶促的调节机制.     

黄河三角洲盐碱土壤中AM真菌的初步调查

2000年3月－2001年2月对黄河三角洲盐碱土壤中5种优势植物柽柳（Tamarix chinensis),芦苇（Phragmites communis),碱蓬（Suaeda glauca),獐毛（Aeluropus littralis var.sinensis) 和刺儿菜（Cirsium setosum)根围内丛枝菌根（arbuscular mycorrhiza,AM)真菌进行了长期定点调查,结果表明,AM真菌在这5种植物根围土壤内都有分布,但多样性较低,尤其在冬季,AM真菌种的丰度和孢子密度分别在夏季降低了18．0％和61．6％,不同深度土层中AM真菌的分布存在诸多差异。     

电子废弃物拆解区土壤重金属污染对丛枝菌根真菌多样性的影响

【背景】电子废弃物拆解造成的土壤重金属污染引发的环境问题日益突出,丛枝菌根(arbuscular mycorrhizal,AM)真菌能侵染植物根系并增强植物抵御环境胁迫的能力,具有重要的生态功能和应用潜力。【目的】探究电子废弃物拆解区土壤重金属污染对AM真菌群落结构与多样性的影响,甄别可耐受重金属污染的AM真菌类群。【方法】从浙江台州某典型电子废弃物拆解场地及其周边区域共采集土壤样品12份,针对土壤中AM真菌的18S rRNA基因进行高通量测序以及可操作分类单元(operational taxonomic unit,OTU)相对丰度和多样性指数计算。【结果】该区土壤中AM真菌由原囊霉目(Archaeosporales)、球囊霉目(Glomerales)和多孢囊霉目(Diversisporales)组成,其中球囊霉目占据优势地位。土壤AM真菌多样性指数与重金属的浓度、综合污染指数和潜在生态风险指数间均无显著相关性,但疑似泡囊根生囊霉(Rhizophagus vesiculiferus)的OTU相对丰度与上述重金属污染指标之间均呈显著正相关关系。【结论】R. vesiculiferus可能对重金属污染有极强耐受性,可为今后电子废弃物拆解污染土壤治理提供技术基础。     

丛枝菌根真菌侵染Bt玉米及对其生长的影响

以2个不同转化事件的Bt玉米5422Bt1(Bt11)和5422CBCL(Mon810)及其同源常规玉米5422为对象,研究接种摩西球囊霉(Glomus mosseae)对Bt玉米与常规玉米生长的影响.结果表明:3个品系玉米接菌处理的菌根侵染率明显不同,菌根侵染各参数值总体呈现5422Bt1>5422CBCL>5422趋势;接种处理均促进了玉米的生长,其效应随玉米品系和生育期的不同而呈现差异,喇叭口期5422的地上部干重和总干重显著增加,拔节期5422Bt1的叶片数、茎粗以及喇叭口期的叶片数、茎粗、地上部干重、总干重显著高于对照,5422CBCL拔节期的叶片数和喇叭口期地下部干重增加显著;3个玉米品系菌根真菌依赖性与接菌效应为5422Bt1>5422CBCL>5422,2个Bt玉米品系与Glomus mosseae间的兼容性更好,接种处理对Bt玉米的生长促进作用强于常规玉米.     

长期肥料施用对华北大田玉米土壤丛枝菌根真菌群落结构的影响

华北平原是我国北方最重要的粮食主产区之一,过去30年间其粮食产量稳定提高的同时,肥料等生产资料的投入也大幅增加。丛枝菌根(Arbuscular mycorrhiza,AM)真菌作为一类重要的土壤功能性微生物,研究施肥措施对其群落、功能的影响,在挖掘农田有益微生物的潜在作用方面具有重要意义。本研究采用分子生物学方法测定了华北平原典型集约化农田体系中不同养分用量和投入模式对土壤AM真菌群落的影响。试验样地于2007年开始对冬小麦-夏玉米轮作体系进行施肥处理:有机肥(牛粪9 000kg/hm~2)、高量化肥(296.4kg N/hm~2、177.3kg P/hm~2、287.8kg K/hm~2,较当地常规化肥施用量上调25%)、低量化肥(74.1kg N/hm~2、44.3kg P/hm~2、71.9kg K/hm~2,较当地常规化肥施用量下调75%)和不施肥对照,每个处理3次重复,于2011年10月玉米收获季进行土壤取样。试验获得的31个OTUs主要属于Glomerales目(球囊霉目)(Glomus Group A/B)及Diversisporales目(多孢囊霉目),其中部分AM真菌类群如Glo12、Aca3、Scutellospora aurigloba(Scut.aurigloba全球盾巨孢囊霉)以及Div2仅在有机肥处理中被检测到。与其他处理相比,有机肥处理的116bp片段(Glo9/12/13/14;Fun1/2;Rhi1)相对丰度显著升高,而140bp片段(Uncultured Glomus)(球囊霉属)则主要受到化肥处理的促进。对于Diversisporales目AM真菌所对应的片段(141、142、169bp),施肥明显对其有促进作用。4年的肥料施用对土壤AM真菌物种多样性无显著影响,施肥处理0–20、20–40、40–60cm深度土壤AM真菌群落的多样性水平分别为1.14–1.78、1.61–1.84、1.23–2.07,丰富度分别为3.7–7.5、8.0–8.3、5.0–7.3。表明深层土壤是AM真菌重要的库,多样性水平并不低,对于维持土壤微生物多样性水平具有重要作用。本研究发现,p H值是影响表层土壤AM真菌群落分布的主要原因,土壤全氮、全碳、速效磷等肥力因子也是影响表层土壤AM真菌群落分布的重要因子,而深层土壤AM真菌的群落组成与固有的盐分特征(土壤电导率EC)有着较好的相关性。     

Community analysis of arbuscular mycorrhizal fungi and bacteria in the maize mycorrhizosphere in a long-term fertilization trial

In this study, we investigated the impact of organic and mineral fertilizers on the community composition of arbuscular mycorrhizal (AM) fungi and bacteria in the mycorrhizosphere of maize in a field experiment established in 1956, in south-east Sweden. Roots and root-associated soil aggregates were sampled four times during the growing season in 2005, in control plots and in plots amended with calcium nitrate, ammonium sulphate, green manure, farmyard manure or sewage sludge. Fungi in roots were identified by cloning and sequencing, and bacteria in soil aggregates were analysed by terminal-restriction fragment length polymorphism, cloning and sequencing. The community composition of AM fungi and bacteria was significantly influenced by the different fertilizers. Changes in microbial community composition were mainly correlated with changes in pH induced by the fertilization regime. However, other factors, including phosphate and soil carbon content, also contributed significantly to these changes. Changes in bacterial community composition and a reduction in bacterial taxon richness throughout the growing season were also manifest. The results of this study highlight the importance and significant effects of the long-term application of different fertilizers on edaphic factors and specific groups of fungi and bacteria playing a key role in arable soils.     

Effects of using different host plants and long-term fertilization systems on population sizes of infective arbuscular mycorrhizal fungi

The effect of cultivation of mycorrhizal and non-mycorrhizal plants and mineral fertilization on the arbuscular mycorrhizal fungal (AMF) community structure of maize (Zea mays L.) plants was studied. Soil samples were collected from two field experiments treated for 5 years with three fertilization systems (Control – no fertilization; Mineral – NPK fertilization; and Organic – Farmyard manure fertilization). Soil samples containing soil and root fragments of rapeseed (Brassica napus L., non-mycorrhizal plant) and wheat (Triticum aestivum L., mycorrhizal plant) collected from the field plots were used as native microbial inoculum sources to maize plants. Maize plants were sown in pots containing these inoculum sources for four months under glasshouse conditions. Colonization of wheat roots by AMF, AMF community structure, AMF diversity (Shannon’s index), AMF dominance (Simpson’s index) and growth of maize were investigated. Sixteen AMF species were identified from rhizosphere soil samples as different species of genera Acaulospora, Claroideoglomus, Dentiscutata, Funneliformis, Gigaspora, Quatunica, Racocetra, and Rhizoglomus. Maize plants grown in manure-fertilized soils had a distinct AMF community structure from plants either fertilized with mineral NPK-fertilizer or non-fertilized. The results also showed that inoculum from non-mycorrhizal plants combined with mineral fertilization decreased AMF diversity (Shannon’s index), AMF dominance (Simpson’s index) and growth of maize. Our findings suggest that non-mycorrhizal plants, such as B. napus, can negatively affect the presence and the effects of soil inoculation on maize growth. Also, our results highlight the importance of considering the long-term effect of rapeseed cultivation system on the reduction of population sizes of infective AMF, and its effect on succeeding annual crops.     

中国植物丛枝菌根侵染特征研究

丛枝菌根真菌在生态系统和植物生长过程中发挥着重要作用,目前对其生理生态功能的研究备受关注,但主要集中在丛枝菌根真菌对植物促生作用方面。植物的菌根侵染特征是表征真菌与植物共生紧密程度的重要标志,也是评价植物生态适应性的关键指标,然而针对我国植物丛枝菌根发育特征及其分布特点的系统性研究尚属空白。【目的】探究我国植物丛枝菌根真菌的侵染特征,及其在不同生态系统和气候区域中的分布规律,为推动丛枝菌根研究的发展提供基础数据。【方法】利用全球植物菌根数据库“FungalRoot”和605篇中文文献的植物菌根侵染特征数据,并补充生态系统、气候区域、植物类型和植物生长年限等共47 700组数据,建立了中国植物丛枝菌根侵染信息数据库,并以此为基础进行研究。【结果】我国植物丛枝菌根真菌侵染率在0-55%占69.7%,55%-100%占30.3%,绝大多数植物丛枝菌根真菌侵染强度、菌丝丰度、泡囊丰度和丛枝丰度均分布在40.0%以下。丛枝菌根共生形态中,A型占比最大,为56.3%。农田、荒漠、草地生态系统的植物丛枝菌根真菌侵染率相近,分别为51.8%、51.6%、51.8%,而森林生态系统的侵染率较低,为40.4%。荒漠生态系统植物的丛枝菌根真菌侵染强度、菌丝丰度、泡囊丰度和丛枝丰度最高,分别达到46.0%、47.1%、37.2%和31.2%。根据气候区域,植物的侵染水平由高到低排序为暖温带(53.3%)>热带(50.0%)>中温带(45.2%)>亚热带(42.2%)。草本植物的侵染水平高于木本植物,多年生植物高于一年生植物。木本植物中,灌木的侵染率水平最高,为46.3%,其次是乔木和藤本植物,均为43.9%。草本植物在侵染强度和菌丝丰度上高于木本植物,分别为30.2%和32.5%,而木本植物在泡囊和丛枝丰度方面具有优势,分别为19.5%和23.4%。我国丛枝菌根植物中,被子植物占据绝大多数,共有110科,占比达到90.2%,蕨类、裸子、石松门植物较少。【结论】我国大部分植物丛枝菌根真菌侵染率在55%以下,侵染特征等指标分布在40.0%以下;同时,不同生态系统、气候区域、植物类型和生长年限均会对侵染特征产生不同程度的影响。     

Effect of arbuscular mycorrhizal fungal inoculation on heavy metal accumulation of maize grown in a naturally contaminated soil

A pot culture experiment was carried out to study heavy metal (HM) phytoaccumulation from soil contaminated with Cu, Zn, Pb, and Cd by maize (Zea mays L.) inoculated with arbuscular mycorrhizal (AM) fungi (AMF). Two AM fungal inocula--MI containing only one AM fungal strain (Glomus caledonium 90036) and MII consisting of Gigaspora margarita ZJ37, Gigaspora decipens ZJ38, Scutellospora gilmori ZJ39, Acaulospora spp., and Glomus spp.--were applied to the soil under unsterilized conditions. The control received no mycorrhizal inoculation. The maize plants were harvested after 10 wk of growth. MI-treated plants had higher mycorrhizal colonization than MII-treated plants. Both MI and MII increased P concentrations in roots, but not in shoots. Neither MI nor MII had significant effects on shoot or root dry weight (DW). Compared with the control, shoot Cu, Zn, Pb, and Cd concentrations were decreased by MI but increased by MII. Cu, Zn, Pb, and Cd uptake into shoots and roots all increased in MII-treated plants, while in MI-treated plants Cu, Zn, and Pb uptake into shoots and Cd uptake into roots decreased but Cu, Zn, and Pb uptake into roots and Cd into shoots increased. MII was more effective than MI in promoting HM extraction efficiencies. The results indicate that MII can benefit HMphytoextraction and, therefore, show potential in the phytoremediation of HM-contaminated soils.     

Effect of arbuscular mycorrhizal fungi, organic fertilizer and soil sterilization on maize growth

The effects of inoculation with arbuscular mycorrhizal (AM) fungi, organic fertilizer (F) applications, and soil sterilization on maize growth were evaluated in a pot experiment. The experiment was in a completely randomized factorial design (2 × 4 × 2) with six replicates for each treatment. There were two soil treatments (sterilized soil, SS and unsterilized soil, US), four organic fertilizer treatments (0.0, 0.5, 1.0 and 2.0 g kg^?1 soil), and two AM fungi treatments (inoculation with Glomus mosseae, +AM and uninoculated control, ?AM). Inoculated plants generally had greater AM colonization, plant height, dry weight and phosphorus (P) uptake than uninoculated controls, and these parameters were significantly increased as the organic fertilizer application increased up to 0.5 g kg^?1 but decreased or had no significant effect compared to the uninoculated plants at the highest fertilizer rate (2.0 g kg^?1). Plant growth, P uptake and AM colonization of root system were significantly higher in sterilized soil compared to the unsterilized control. Our results indicated that the inoculation of AM fungi in field soil with optimal organic fertilizer application greatly improved maize growth and nutrient uptake, and the effect was greater under sterilized soil condition.     

Effect of arbuscular mycorrhizal fungi, organic fertilizer and soil sterilization on maize growth ☆

The effects of inoculation with arbuscular mycorrhizal (AM) fungi, organic fertilizer (F) applications, and soil sterilization on maize growth were evaluated in a pot experiment. The experiment was in a completely randomized factorial design (2 × 4 × 2) with six replicates for each treatment. There were two soil treatments (sterilized soil, SS and unsterilized soil, US), four organic fertilizer treatments (0.0, 0.5, 1.0 and 2.0 g kg^-1 soil), and two AM fungi treatments (inoculation with Glomus mosseae, +AM and uninoculated control, －AM). Inoculated plants generally had greater AM colonization, plant height, dry weight and phosphorus (P) uptake than uninoculated controls, and these parameters were significantly increased as the organic fertilizer application increased up to 0.5 g kg^-1 but decreased or had no significant effect compared to the uninoculated plants at the highest fertilizer rate (2.0 g kg^-1). Plant growth, P uptake and AM colonization of root system were significantly higher in sterilized soil compared to the unsterilized control. Our results indicated that the inoculation of AM fungi in field soil with optimal organic fertilizer application greatly improved maize growth and nutrient uptake, and the effect was greater under sterilized soil condition.     

长期定位施肥对石灰性紫色水稻土AMF多样性的影响

为了解长期定位施肥对石灰性紫色水稻土丛枝菌根真菌（AMF）物种多样性的影响,从稻麦轮作下连续26年定位施农家肥（M）、氮肥+农家肥（NM）、氮磷肥+农家肥（NPM）、氮磷钾肥+农家肥（NPKM）、不施肥（CK）、氮肥（N）、氮磷肥（NP）、氮磷钾肥（NPK）等不同施肥处理小区采集根区土样,分离鉴定AMF、测定多样性特征。共鉴定出17种AMF,其中球囊霉属Glomus 12种,占已分离总数的70.5%,为优势属;无梗囊霉属Acaulospora 3种,占17.6%;原囊霉属Archaeospora和内养囊霉属Entrophospora各1种,各占5.9%。无论是石灰性紫色土种植水稻还是小麦,AMF侵染率最高的施肥处理均为无肥处理（CK）,施肥特别是磷肥,降低AMF侵染率、孢子密度、相对多度和种丰度。变性梯度凝胶电泳（denaturing gradient gel electrophoresis,DGGE）分析结果表明,不施肥对照（CK）的AMF多样性最丰富,多样性指数最高,水稻和小麦种植下的多样性指数分别为2.68和3.02;与无机肥处理相比,农家肥配施无机肥提高了AMF多样性指数。农家肥配施无机肥更有利于AMF发育。     

丛枝菌根真菌对砂培玉米幼苗根系特征、光合生理与镉累积的影响

【背景】丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)是一类重要的土壤微生物,能显著影响植物对镉(cadmium,Cd)的耐性与累积,但其对不同形态Cd胁迫的响应尚不清楚。【目的】探讨不同形态Cd胁迫下接种AMF对玉米(Zea mays L.)生长和Cd累积的影响。【方法】采用30 cm高的培养容器填装石英砂(0.2 mm),开展室内砂培玉米试验,研究溶解态和胶体态Cd (1 mg/kg)胁迫下,接种摩西斗管囊霉(Funneliformis mosseae)对玉米幼苗生长、根系特征、光合生理及Cd累积的影响。【结果】双因素分析表明,AMF和Cd形态均对玉米生长(株高和生物量)、根系特征、光合生理(叶绿素含量和光合速率)与Cd累积量存在显著的影响,但二者之间没有显著交互作用。与未接种处理相比,接种AMF显著降低玉米株高、生物量、叶片叶绿素含量和光合速率,抑制玉米根长、根表面积、根体积和根尖数;同时增加了玉米根系Cd含量,但减少玉米地上部Cd含量以及地上部与根系Cd累积量;与胶体态Cd处理相比,溶解态Cd显著降低玉米的根长、根表面积、平均根系直径、根尖数和地上部Cd累积量,但增加了植株叶片光合速率、根系Cd含量和累积量。相关分析发现,玉米根长、根表面积和根尖数与地上部Cd含量呈显著或极显著正相关,与根系Cd含量呈极显著负相关。【结论】溶解态Cd比胶体态Cd对砂培玉米幼苗的毒害效应严重,而且接种AMF加重溶解态和胶体态Cd对玉米幼苗的损伤,但降低了植株对Cd的累积。     

Selection of appropriate host plants used in trap culture of arbuscular mycorrhizal fungi

Arbuscular mycorrhizal (AM) fungi in coalmine spoil, island forest and saline soils were enriched in pot culture with maize (Zea mays L.), tobacco (Nicotiana tabacum L.), white clover (Trifolium repens Linn.) and silverweed cinquefoil (Potentilla anserina L.). Based on spores, there were more species of AM fungi in the coalmine spoil (15 species, 3 genera), than in the forest soil (11 species, 4 genera) and the saline soil (5 species, 2 genera). In the trap cultures, the total of 28 species in Acaulospora, Gigaspora, Glomus, and Sclerocystis detected in the original soils were all recovered with at least one of the four trap plants. The highest spore and species numbers were recovered in trap cultures of T. repens inoculated with coalmine spoil. Glomus constrictum and Glomus multicaule were the dominant species associated with N. tabacum grown in saline soil and forest soil. The dominant species of AM fungi on the four hosts was Acaulospora mellea, which had over 90% of the spore incidence in pot trap culture in coalmine spoil. It is suggested that there be selectivity between host plants and AM fungi. The number of species of AM fungi detected was influenced by host plants under certain conditions and white clover was generally the optimal host plant to detect diversity of AM fungi.