城市生态系统中丛枝菌根真菌侵染状况及群落特征

作者采集了北京、青岛、济南、南京、武汉和贵阳6个城市中部分大学校园高羊茅Festuca elata、城内公园大叶黄杨Euonymus japonicus及郊区公园连翘Forsythia suspensa根区土样,测定菌根着生状况和丛枝菌根（arbuscular mycorrhizal,AM）AM真菌孢子密度,形态鉴定AM真菌种类,测定AM真菌种丰度、多样性指数及群落结构等。除从北京中国人民大学校园草坪和南京下马坊公园的大叶黄杨根系上未见丛枝结构外,其他根系样品均观测到典型的丛枝和泡囊结构,丛枝着生率较低,且多为A-型。各城市大多数样品的菌根总侵染率、丛枝着生率和泡囊数分别低于40%、5%和1/mm。北京中国人民大学草坪根系菌根总侵染率显著低于其他城市校区的。除北京香山的连翘和济南山东大学的高羊茅上的种丰度、Shannon指数、以及北京紫竹院公园大叶黄杨上的孢子密度之外,青岛市的3种植物上AM真菌的孢子密度、种丰度和Shannon指数均高于其他城市的。各校园草坪中相同的AM真菌种类较少,但放射球囊霉Glomus radiatum分布频率100%,为校园草坪中的优势种。城市公园以青岛中山公园的AM真菌的孢子密度和种丰度最高;郊区公园以贵阳黔灵山的孢子密度、种丰度和Shannon指数最高,武汉狮子山的种丰度和Shannon指数最低。土壤氮沉降与泡囊数量呈显著负相关,土壤压实程度与泡囊数量呈显著正相关。大叶黄杨根区土壤中AM真菌种丰度与土壤有机质含量呈显著正相关,Shannon指数与土壤pH呈显著正相关。结论认为,中国不同城市生态系统中AM真菌群落结构差异较大,氮沉降和土壤压实程度对根内泡囊数量具有不同的影响。     

Microbial inoculation influences arbuscular mycorrhizal fungi community structure and nutrient dynamics in temperate tree restoration

Soil microbial communities have a profound influence on soil chemical processes and subsequently influence tree nutrition and growth. This study examined how the addition of a commercial inoculum or forest‐collected soils influenced nitrogen (N) and phosphorus (P) dynamics, soil microbial community structure, and growth in Liriodendron tulipifera and Prunus serotina tree saplings. Inoculation method was an important determinant of arbuscular mycorrhizal fungi (AMF) community structure in both species and altered soil N dynamics in Prunus and soil P dynamics in Liriodendron. Prunus saplings receiving whole forest soil transfers had a higher rhizosphere soil carbon/nitrogen ratio and ammonia content at the end of the first growing season when compared to unmanipulated control saplings. Inoculation with whole forest soil transfers resulted in increased inorganic phosphorus in Liriodendron rhizosphere soils. The number of AMF terminal restriction fragments was significantly greater in rhizosphere soils of Liriodendron saplings inoculated with whole forest soil transfers and Prunus saplings receiving either inoculum source than control saplings. Forest soil inoculation also increased AMF colonization and suppressed stem elongation in Liriodendron after 16 months; conversely, Prunus AMF colonization was unchanged and stem elongation was significantly greater when saplings were inoculated with whole forest soil transfers. Longer term monitoring of tree response to inoculation will be essential to assess whether early costs of AMF colonization may provide long‐term benefits. This study provides insight into how practitioners can use microbial inoculation to alter AMF community structure and functioning, subsequently influencing tree growth and nutrient cycling during the restoration of degraded lands.     

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。     

长期定位施肥土壤中AM真菌对寄主植物的侵染状况

分别于2003年9月和2004年5月在莱阳农学院长期(26年)定位施肥试验田采集玉米和小麦根系,以测定不同施肥处理对丛枝菌根(AM)真菌侵染状况的影响。结果表明长期定位施氮(N)肥显著降低了AM真菌对寄主植物的侵染率(MCP)、丛枝着生率(ACP)、单位根长泡囊数(NV)和侵入点数(NE)。高N处理的小麦MCP低于低N处理,高N处理的玉米ACP低于低N处理。长期定位施有机肥,尤其是高有机肥处理显著降低了MCP、ACP和NE。有机肥和N肥配施也降低了MCP、ACP、NV和NE,以高有机肥和高N肥配施处理的降低效应最大。除小麦根系NV外,长期定位NPK配合施用降低了玉米和小麦的MCP、ACP、NV和NE。其中,以NP处理的玉米ACP、小麦的MCP和ACP最低。另外,低有机肥与N肥配施和NPK配施处理条件下玉米MCP高于小麦。结论认为不同施肥体制对作物菌根生长发育及其结构具有不同影响,而且有机肥和N肥在影响AM真菌侵染方面存在互作。     

Interactive effects of root endophytes and arbuscular mycorrhizal fungi on an experimental plant community

Plant-soil microbial interactions have moved into focus as an important mechanism for understanding plant coexistence and composition of communities. Both arbuscular mycorrhizal (AM) as well as other root endophytic fungi co-occur in plant roots, and therefore have the potential to influence relative abundances of plant species in local assemblages. However, no study has experimentally examined how these key root endosymbiont groups might interact and affect plant community composition. Here, using an assemblage of five plant species in mesocosms in a fully factorial experiment, we added an assemblage of AM fungi and/or a mixture of root endophytic fungal isolates, all obtained from the same grassland field site. The results demonstrate that the AM fungi and root endophytes interact to affect plant community composition by changing relative species abundance, and consequently aboveground productivity. Our study highlights the need to explicitly consider interactions of root-inhabiting fungal groups in studies of plant assemblages.     

High diversity of arbuscular mycorrhizal fungi in a boreal herb-rich coniferous forest

* Here, the diversity of arbuscular mycorrhizal (AM) fungi was determined in a boreal herb-rich coniferous forest in relation to environmental variables. * Root samples of five plant species (Fragaria vesca, Galeobdolon luteum, Hepatica nobilis, Oxalis acetosella and Trifolium pratense) were analysed from stands differing in age and forest management intensity. * Thirty-four Glomeromycota taxa (small-subunit ribosomal RNA gene (SSU rDNA) sequence groups) were detected from 90 root samples (911 clones), including eight new taxa. Sequence groups related to Glomus intraradices were most common (MO-G3 and MO-G13). Samples of H. nobilis were colonized by more AM fungal taxa (3.68 +/- 0.31) than those of O. acetosella (2.69 +/- 0.34), but did not differ significantly in this respect from those of F. vesca (3.15 +/- 0.38). Effects of forest management, host plant species (except above) or season on the number or composition of fungal taxa in root samples were not detected, and neither were they explained by environmental variables (vegetation, soil and light conditions). * This is the most taxon-rich habitat described to date in terms of root-colonizing Glomeromycota. The data demonstrate the importance of temperate coniferous forests as habitats for AM fungi and plants. Lack of obvious fungal community patterns suggests more complex effects of biotic and abiotic factors, and possibly no adverse effect of common forest management practices on AM fungal diversity.     

Simulated nitrogen deposition affects community structure of arbuscular mycorrhizal fungi in northern hardwood forests

Our previous investigation found elevated nitrogen deposition caused declines in abundance of arbuscular mycorrhizal fungi (AMF) associated with forest trees, but little is known about how nitrogen affects the AMF community composition and structure within forest ecosystems. We hypothesized that N deposition would lead to significant changes in the AMF community structure. We studied the diversity and community structure of AMF in northern hardwood forests after more than 12 years of simulated nitrogen deposition. We performed molecular analyses on maple (Acer spp.) roots targeting the 18S rDNA region using the fungal‐specific primers AM1 and NS31. PCR products were cloned and identified using restriction fragment length polymorphism (RFLP) and sequencing. N addition significantly altered the AMF community structure, and Glomus group A dominated the AMF community. Some Glomus operational taxonomic units (OTUs) responded negatively to N inputs, whereas other Glomus OTUs and an Acaulospora OTU responded positively to N inputs. The observed effect on community structure implies that AMF species associated with maples differ in their response to elevated nitrogen. Given that functional diversity exists among AMF species and that N deposition has been shown to select less beneficial fungi in some ecosystems, this change in community structure could have implications for the functioning of this type of ecosystem.     

Cryopreservation of arbuscular mycorrhizal fungi from root organ and plant cultures

Long-term maintenance of arbuscular mycorrhizal fungi (AMF) by in vitro or in vivo subcultivation is often expensive and time-consuming and could present the risk of contaminations and possibly morphological, physiological, and genetic variations over time. Recently, in vitro produced AMF isolates belonging to the genus Rhizophagus were successfully cryopreserved at ?130 °C following encapsulation-drying. Here, this method was tested on 12 single species cultures belonging to six different genera (i.e., Rhizophagus, Glomus, Claroideoglomus, Septoglomus, Paraglomus, and Gigaspora) produced in vitro or in vivo. Their viability was estimated, after 1 month of cryopreservation at ?130 °C, by the percentage of potentially infective beads (i.e., the percentage of beads that contained at least one germinated propagule) for the in vitro produced species and the percentage of infective beads (i.e., the percentage of beads that contained at least one propagule able to colonize a new host plant in pot culture) for the in vivo produced species. With the exception of Gigaspora sp. MUCL 52331 and Septoglomus constrictus PER 7.2, no significant differences were observed in the viability of the single species cultures before and after cryopreservation. These results, thus, demonstrated the suitability of the cryopreservation method by encapsulation-drying for AMF species belonging to different genera and produced in vitro or in vivo. This method opens the door to the long-term preservation at ultra-low temperature of a large number of AMF species and for the preservation of species that are still recalcitrant to in vitro cultivation.     

不同品种牡丹对丛枝菌根真菌群落结构的影响

对山东菏泽赵楼牡丹园栽培的不同品种牡丹(Paeonia suffruticosa)根围丛枝菌根(ar-buscular mycorrhiza,AM)真菌孢子密度、种属构成、种丰度、分布频度、物种多样性指数及其物种组成相似性等进行了研究.结果表明:不同品种牡丹根围AM真菌的种属构成、种丰度和分布频度等均不相同,其中,从‘凤丹'和‘赵粉'根围中各分离到10种AM真菌,从‘乌龙捧盛'和‘洛阳红'根围中分离到9种,从‘胡红'根围中分离到8种;‘凤丹'根围AM真菌孢子密度最高,为59个·50 g-1土,‘胡红'最低,为47个·(50 g)-1土;‘赵粉'的物种多样性指数最高(1.89),‘胡红'最低(1.71).‘凤丹'和‘胡红'的菌根侵染率最高,为63.6%,‘乌龙捧盛'最低,为52.7%.不同牡丹品种之间AM真菌种类组成的相似性系数在0.71～0.95,其中‘乌龙捧盛'和‘凤丹'的相似性系数最高(0.95),而‘胡红'与‘洛阳红'的相似性系数最低(0.71).牡丹基因型能改变AM真菌群落结构特征.     

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.     

Colonization of pennycresses (Thlaspi spp.) of the Brassicaceae by arbuscular mycorrhizal fungi

Members of the Brassicaceae are generally believed to be non-mycorrhizal. Pennycress (Thlaspi) species of this family from diverse locations in Slovenia, Austria, Italy and Germany were examined for their colonisation by arbuscular mycorrhizal fungi (AMF). Meadow species (T. praecox, T. caerulescens and T. montanum) were sparsely but distinctly colonised, as indicated by the occurrence of intraradical hyphae, vesicles, coils, and occasionally arbuscules. Species from other locations were poorly colonised, but arbuscules were not discernible. The genus Thlaspi comprises several heavy metal hyperaccumulating species (T. caerulescens, T. goesingense, T. calaminare, T. cepaeifolium). All samples collected from heavy metal soils were at best poorly colonized. Thus the chance is small to find a "hypersystem" in phytoremediation consisting of an AM fungus which prevents the uptake of the major part of the heavy metals and of a Thlaspi species which effectively deposits the residual heavy metals inevitably taken up into its vacuoles. In two different PCR approaches, fungal DNA was amplified from most of the Thlaspi roots examined, even from those with a very low incidence of AMF colonization. Sequencing of the 28S- and 18S-rDNA PCR-products revealed that different Thlaspi field samples were colonized by Glomus intraradices and thus by a common AM fungus. However, none of the sequences obtained was identical to any other found in the present study or deposited in the databanks, which might indicate that a species continuum exists in the G. intraradices clade. An effective colonization of Thlaspi by AMF could not be established in greenhouse experiments. Although the data show that Thlaspi can be colonized by AMF, it is doubtful whether an effective symbiosis with the mutual exchange of metabolites is formed by both partners.     

Nested multiplex PCR—A feasible technique to study partial community of arbuscular mycorrhizal fungi in field-growing plant root

Plant can be infected by different arbuscular mycorrhizal fungi, but little is known about the interaction between them within root tissues mainly because different species cannot be distinguished on the basis of fungal structure. Accurate species identification of Arbuscular mycorrhizal fungi (AMF) colonized in plant roots is the comerstone of mycorrhizal study, yet this fundamental step is impossible through its morphological character alone. For accurate, rapid and inexpensive detection of partial mycorrhizal fungal community in plant roots, a nested multiplex polymerase chain reaction (PCR) was developed in this study. Five discriminating primers designed based on the variable region of the 5′ end of the large ribosomal subunit were used in the experiment for testing their specificity and the sensitivity in nested PCR by using spores from Glomus mosseae (BEG12), Glomus intraradices (BEG141), Scutellospora castaneae (BEG1) and two unidentified Glomus sp. HAUO3 and HAUO4. The feasibility assay of nested multiplex PCR was conducted by use of spore mixture, Astragalus sinicum roots co-inoculated with 4 species of arbuscular mycorrhizal fungi from pot cultures and 15 different field-growing plant roots respectively after analyses of the compatibility of primers. The result indicated that the sensitivity was in the same range as that of the corresponding single PCR reaction. Overall accuracy was 95%. The efficiency and sensitivity of this multiplex PCR procedure provided a rapid and easy way to simultaneously detect several of arbuscular mycorrhizal fungal species in a same plant root system.     

丛枝菌根真菌形态结构、物种多样性和群落组成对氮沉降响应研究进展

工业革命以来,人类活动输入到生态系统中的氮迅速增加,已突破地球所能承受的氮循环阈值。过量氮沉降会造成生物多样性丧失等一系列危害,严重影响生态系统结构和功能。丛枝菌根(AM)真菌能够与大约70%-80%的陆地植物种类形成共生关系,在宿主植物养分吸收、抵抗外界不良环境压力、群落动态和物种共存、生物地球化学循环等方面具有重要的作用。探究AM真菌对氮沉降的响应对认识和把握菌根真菌缓解氮沉降的负面后果,维持生态系统的结构和功能具有重要意义。本文综述了AM真菌的形态结构、物种多样性和群落组成等对氮沉降的响应机制。前人研究表明氮沉降通常降低AM真菌的根系定殖率,减少根外菌丝密度和土壤孢子密度,改变菌丝生长的时间动态;降低AM真菌多样性,改变AM真菌群落组成。氮沉降主要通过缓解植物氮限制、降低植物对菌根的依赖性、减少植物对菌根的碳分配、改变根系和土壤中菌根生物量比率、在植物根内维持稳定的菌根真菌组成作为应对未来扰动的“保险”、改变土壤资源有效性及土壤酸度等直接和间接途径影响AM真菌结构和功能。我们建议在未来研究中整合多组学手段、开展学科交叉,聚焦复杂的生物互作体系对氮沉降的响应机制,以及AM真菌对氮沉降响应的生态后果。     

荒漠北沙柳根系丛枝菌根真菌和黑隔内生真菌定殖状况

为利用土壤共生真菌资源促进荒漠植被恢复和生态重建, 分别于2013年6月、8月和10月, 从内蒙古元上都地区采集北沙柳(Salix psammophila)根围0-10、10-20、20-30、30-40和40-50 cm共5个土层的土壤样品, 系统研究了丛枝菌根真菌(AMF)和黑隔内生真菌(DSE)的时空分布及其与土壤因子的相关性。结果表明: AMF和DSE的平均定殖率分别为77%和84%, 说明北沙柳根系能与这两类真菌形成良好的共生关系。AMF和DSE的分布和定殖具有明显的时空异质性, 并与土壤因子密切相关。AMF和DSE的平均定殖率均表现为10月> 8月> 6月。土壤深度对AMF和DSE的定殖率有显著影响, AMF和DSE定殖率的最大值分别在0-20 cm和0-10 cm土层。双因子方差分析表明, 月份和土层对AMF和DSE的定殖率以及土壤因子具有显著的交互效应。主成分分析表明, 土壤湿度、pH值、碱性磷酸酶、易提取球囊霉素是内蒙古荒漠环境中AMF和DSE定殖的主要影响因子。     

Temperature constraints on the growth and functioning of root organ cultures with arbuscular mycorrhizal fungi

In this study we investigated the effects of temperature on fungal growth and tested whether the differences in fungal growth were related to the effects of temperature on carbon movement to, or within, the fungus. Growth curves and C uptake-transfer-translocation measurements were obtained for three arbuscular mycorrhizal fungi (AMF) isolates cultured within a 6-30 degrees C temperature range. A series of experiments with a model fungal isolate, Glomus intraradices, was used to examine the effects of temperature on lipid body and 33P movement, and to investigate the role of acclimation and incubation time. Temperature effects on AMF growth were both direct and indirect because, despite clear independent root and AMF growth responses in some cases, the uptake and translocation of 13C was also affected within the temperature range tested. Root C uptake and, to a lesser extent, C translocation in the fungus, were reduced by low temperatures (< 18 degrees C). Uptake and translocation of 33P by fungal hyphae were, by contrast, similar between 10 and 25 degrees C. We conclude that temperature, between 6 and 18 degrees C, reduces AMF growth, and that C movement to the fungus is involved in this response.     

Inoculation with arbuscular mycorrhizal fungi: the status quo in Japan and the future prospects

Inoculation of arbuscular mycorrhizal (AM) fungi has potential benefits in not only sustainable crop production but also environmental conservation. However, the difficulty of inoculum production due to the obligate biotrophic nature of AM fungi has been the biggest obstacle to putting inoculation into practice. Nevertheless, several companies have sought to produce inoculum of AM fungi. Firstly in this review, the present status of inoculum production and its use in Japan is described. Secondly, although the effectiveness of inoculation is primarily limited by environmental and biological factors, some possible ways to improve inoculation performance are discussed. Approaches include use of chemicals to increase spore germination and colonization and soil application of charcoal to provide a microhabitat for AM fungi to colonize and survive.