丛枝菌根真菌系统分类及群落研究技术进展

丛枝菌根真菌（AMF）是自然生态系统重要的组成部分,能与植物根系形成互惠共生体.传统的AMF分类主要依赖于对土壤无性孢子的形态鉴定,具有一定的局限性.近年来基于核酸分析的分子鉴定技术使AMF的分类更具科学性和准确性,补充和完善了基于孢子形态鉴定所建立的分类系统.AMF群落研究依赖于AMF的分类鉴定,主要包括孢子形态鉴定和分子生物学分析两类研究法.本文综述了AMF的分类系统和群落研究方法,着重介绍了近年来应用较多的AMF群落研究的分子生物学技术.作者认为,采取形态与分子相结合的办法将有助于推动AMF群落研究和AMF自然分类系统的建立和完善.     

Genetic variation and species identification among selected leeches (Hirudinea) revealed by RAPD markers

Leech taxonomy is based on unstable morphological characters. The overall level of genetic variability and species differentiation is unknown. Using the RAPD assays genetic diversities and genetic similarities were estimated in twelve species collected in North-Eastern Poland and representing three families and five subfamilies. Ten primers revealed 204 reproducible bands. Genetic diversities varied from 0.099 to 0.219 classifying studied species among variable invertebrates. Total 45 markers comprised 22% of all amplified bands were unique for species thus enabling their identification. Genetic similarities among species (0.528–0.811) evidenced several stages of differentiation, which is mirrored in the current taxonomy. The UPGMA and multidimensional scaling (nMDS) based on our RAPDs are congruent and reflected traditional division into “Rhynchobdellida” and Arhynchobdellida. The RAPD approach proved to be an effective tool in population and evolutionary studies of leeches. For the first time, genetic parameters were estimated enabling to compare leeches with outcomes from other animals.     

Small-scale spatial heterogeneity of arbuscular mycorrhizal fungal abundance and community composition in a wetland plant community

Although it has become increasingly clear that arbuscular mycorrhizal fungi (AMF) play important roles in population, community, and ecosystem ecology, there is limited information on the spatial structure of the community composition of AMF in the field. We assessed small-scale spatial variation in the abundance and molecular diversity of AMF in a calcareous fen, where strong underlying environmental gradients such as depth to water table may influence AMF. Throughout an intensively sampled 2 × 2 m plot, we assessed AMF inoculum potential at a depth of 0–6 and 6–12 cm and molecular diversity of the AMF community using terminal restriction fragment length polymorphism of 18S rDNA. Inoculum potential was only significantly spatially autocorrelated at a depth of 6–12 cm and was significantly positively correlated with depth to water table at both depths. Molecular diversity of the AMF community was highly variable within the plot, ranging from 2–14 terminal restriction fragments (T-RFs) per core, but the number of T-RFs did not relate to water table or plant species richness. Plant community composition was spatially autocorrelated at small scales, but AMF community composition showed no significant spatial autocorrelation. Saturated soils of calcareous fens contain many infective AMF propagules and the abundance and diversity of AMF inoculum is patchy over small spatial scales. An erratum to this article can be found at     

Does the sequence of plant dominants affect mycorrhiza development in simulated succession on spoil banks?

The aim of this field study was to examine how the development of arbuscular mycorrhizal fungi (AMF) on coal mine spoil banks is affected by the presence of plants with different mycorrhizal status. A 3-year trial was conducted on the freshly created spoil bank Vršany, North-Bohemian coal basin, the Czech Republic. Three plant species – non-mycotrophic annual Atriplex sagittata, highly mycotrophic annual Tripleurospermum inodorum (both dominants of early stages of succession) and facultatively mycotrophic Arrhenatherum elatius (a perennial grass species of the later stage of succession) – were planted on 1 m² plots over 3 years in different sequences that simulated the progress of succession on spoil banks. The development of AMF populations was monitored by evaluation of mycorrhizal colonization of plant roots and by measurement of the mycorrhizal inoculation potential (MIP) of soil. These two parameters were compared between plots inoculated with the mixture of three AMF isolates – Glomus mosseae BEG95, G. claroideum BEG96 and G. intraradices BEG140 – (“inoculated plots”) and plots exposed only to natural dispersal of AMF propagules (“uninoculated plots”). Highly colonized roots of plants together with a high MIP of soil in uninoculated plots were already found at the end of the first season, indicating rapid natural dispersal of AMF propagules. Root colonization of facultatively mycotrophic and non-mycotrophic plants in later years was affected by the mycorrhizal status of the previous plant species. The MIP of soil continuously increased throughout the experiment; in uninoculated plots, the MIP was temporarily decreased if plant species of higher mycotrophy were replaced by species of lower mycotrophy. The results lead to the conclusion that AMF colonize freshly formed sites very quickly and reproduce or accumulate in the soil, which leads to increasing MIP values. However, this infective potential can be decreased if non-mycotrophic plants predominate on the site.     

Diversity of arbuscular mycorrhizal fungi in greenhouse soils continuously planted to watermelon in North China

In North China, watermelon is grown in commercial greenhouses in a continuous monoculture and with high application rates of manure or compost. The aim of this study was to determine how the diversity of arbuscular mycorrhizal fungi (AMF) in these soils changed over long periods (0 to 20 years) of monoculture. AMF in control soils (from fields not replanted with watermelon and located near the greenhouses) and in greenhouses (in Daxing, Beijing, and Weifang, Shandong) that had been continuously replanted with watermelon for 5, 10, 15, or 20 years (three greenhouses per year per location) were identified and quantified based on spore morphology and on denaturing gradient gel electrophoresis (DGGE). The total number of AMF species and genera were 13 and 3 in soils replanted for 5–20 years and 19 and 4 in control soils. AMF species richness (SR), the Shannon–Wiener index (H), and spore density declined as the number of years in which watermelon was replanted increased. The available phosphorus, potassium, and nitrogen in the soil increased as the number of years in which watermelon was replanted increased. Values for SR and H were higher when based on DGGE than on spore morphology. The results suggest that current greenhouse practices in North China reduce the AMF diversity in the soil.     

巨孢囊霉目丛枝菌根真菌的分类研究进展

巨孢囊霉目(Gigasporales)真菌因其形态特征显著而被认为是较易归类和鉴定的一类丛枝菌根真菌(AMF)。近十余年来随着研究技术的进步,基于无性孢子形态特征而建立的AMF分类、鉴定方法得以不断补充和完善,也使得巨孢囊霉目真菌的系统分类一直处于不断变化和更新的状态。文中主要介绍了该目真菌基于形态特征的鉴定方法上的最新变化以及最新的分类系统,着重归纳已报道科、属、种分类特征,旨在为开展我国AMF资源调查、丰富其多样性提供便利。     

Promiscuous arbuscular mycorrhizal symbiosis of yam (Dioscorea spp.), a key staple crop in West Africa

Yam (Dioscorea spp.) is a tuberous staple food crop of major importance in the sub-Saharan savannas of West Africa. Optimal yields commonly are obtained only in the first year following slash-and-burn in the shifting cultivation systems. It appears that the yield decline in subsequent years is not merely caused by soil nutrient depletion but might be due to a loss of the beneficial soil microflora, including arbuscular mycorrhizal fungi (AMF), associated with tropical “tree-aspect” savannas and dry forests that are the natural habitats of the wild relatives of yam. Our objective was to study the AMF communities of natural savannas and adjacent yam fields in the Southern Guinea savanna of Benin. AMF were identified by morphotyping spores in the soil from the field sites and in AMF trap cultures with Sorghum bicolor and yam (Dioscorea rotundata and Dioscorea cayenensis) as bait plants. AMF species richness was higher in the savanna than in the yam-field soils (18–25 vs. 11–16 spp.), but similar for both ecosystems (29–36 spp.) according to the observations in trap cultures. Inoculation of trap cultures with soil sampled during the dry season led to high AMF root colonization, spore production, and species richness (overall 45 spp.) whereas inoculation with wet-season soil was inefficient (two spp. only). The use of D. cayenensis and D. rotundata as baits yielded 28 and 29 AMF species, respectively, and S. bicolor 37 species. AMF root colonization, however, was higher in yam than in sorghum (70–95 vs. 11–20%). After 8 months of trap culturing, the mycorrhizal yam had a higher tuber biomass than the nonmycorrhizal controls. The AMF actually colonizing D. rotundata roots in the field were also studied using a novel field sampling procedure for molecular analyses. Multiple phylotaxa were detected that corresponded with the spore morphotypes observed. It is, therefore, likely that the legacy of indigenous AMF from the natural savanna plays a crucial role for yam productivity, particularly in the low-input traditional farming systems prevailing in West Africa.     

Zn uptake by maize under the influence of AM-fungi and Collembola Folsomia candida

Zn uptake by maize plants may be affected by the presence of arbuscular mycorrhizal fungi (AMF). Collembola often play an important controlling role in the inter-relationship between AMF and host plants. The objective of this experiment was to examine whether the presence of Collembola at different densities (0.4 and 1 individuals g⁻¹ dry soil) and their activity have any effect on Zn uptake by maize through the plant–AMF system. The presence of the AMF (Glomus intraradices) and of the Collembola species Folsomia candida was studied in a laboratory microcosm experiment, applying a Zn exposure level of 250 mg kg⁻¹ dry soil. Biomass and water content of the plants were no different when only AMF or when both AMF and Collembola were present. In the presence of AMF the Zn content of the plant shoots and roots was significantly higher than without AMF. This effect was reduced by Collembola at both low and high density. High densities of Collembola reduced the extent of AMF colonization of the plant roots and hyphal length in the soil, but low densities had no effect on either. The results of this experiment reveal that the F. candida–G. intraradices interaction affects Zn uptake by maize, but the mechanisms are still unknown.     

Assessing diversity of arbuscular mycorrhizal fungi in a local community: role of sampling effort and spatial heterogeneity

Diversity of arbuscular mycorrhizal fungi (AMF) was assessed in two 9.2 × 9.2-m plots planted with landscape trees and shrubs at an experimental site in Phoenix, AZ, USA. Twenty-five soil samples were collected in a regular grid pattern from each plot, and AMF species were identified using trap cultures. A total of 12 species were detected, with 7 species detected in one plot and 11 in the other. We found that sampling effort had a major impact on assessing species richness and composition in this local community. Fifteen samples would be necessary to detect 70–80% of species present in each plot. A limited number of additional undetected species are likely to be present in both plots, based on the sampling effort curves and jackknife estimates. Only two species, Glomus eburneum and Glomus microaggregatum, were detected in over 50% of the samples from both plots, and rank–frequency plots revealed a lognormal species distribution. Despite the patchiness of plants in the plots, the number of species detected per point exhibited spatial structuring only at the smallest sampling scale in a single plot, and only a single species in each plot was not randomly distributed. These results indicate that sampling effort and strategy can affect perceptions of AMF community structure.     

Phylogenetic relationships among deer in China derived from mitochondrial DNA cytochromeb sequences

The phylogenetic relationships of Cervidae and Moschidae were examined using partial sequence data of mitochondrial DNA (mtDNA) cytochromeb. Ten new sequences were obtained for six species of Cervidae and Moschidae, and aligned with those previously reported for other deer species. Our results demonstrated that the phylogenetic status of the taxa inferred from molecular data was congruent with taxonomy based on morphological studies. Cervidae formed a monophyletic group that consists of four subfamilies: Cervinae, Muntiacinae, Hydropotinae, and Odocoileinae. Moschidae occurred at the base of the Cervidae clade. On the basis of molecular clocks for genetic distance, the divergence time of mtDNA haplotypes within the subfamily Cervinae, among subfamilies in Cervidae, and between Moschidae and Cervidae was estimated to date 2–7 MYA, 6–10 MYA and 8–13 MYA, respectively.     

丛枝菌根真菌物种多样性研究进展

丛枝菌根真菌(Arbuscular mycorrhizal fungi,AMF)在不同生态系统均发挥至关重要的作用,研究其多样性能够为AMF物种资源的保护和利用提供科学依据。AMF不能被离体纯培养以及自身的高变异性等因素严重阻碍了对其进行深入研究,随着研究方法的不断改进,尤其是新一代测序技术的运用,极大加速了人们对AMF物种多样性的认识。本文主要从AMF分类系统、不同宿主植物和不同生境中的AMF物种多样性及AMF物种多样性研究方法(包括形态鉴定、Sanger测序和高通量测序)方面介绍AMF物种多样性研究进展,并且探讨AMF物种多样性研究中存在的主要问题,认为在今后AMF物种多样性研究中不仅要注重运用新的研究手段,还应该着重解决AMF不能离体纯培养的问题。     

核糖体rDNA序列分析在丛枝菌根真菌研究中的应用

丛枝菌根真菌(AMF)是一类古老、专性活体营养的共生菌物, 尚未获得纯培养, 在一定程度上限制了人们对AMF的深入研究。以DNA分析技术为基础的分子生物学技术增加了AMF检测的敏感性与特异性, rDNA序列的同源性和变化性可更真实地反映物种之间的亲缘关系及进化地位, 因而被广泛应用于AMF分类、鉴定、遗传、生态及物种多样性等研究中。本文简要综述了rDNA序列分析技术在AMF系统发育、分子检测及群落结构特征研究中的应用现状。     

Optimizing tillage schedule for maintaining activity of the arbuscular mycorrhizal fungal population in a rainfed upland rice (Oryza sativa L.) agro-ecosystem

Rainfed uplands in India are predominantly mono-cropped with rice (Oryza sativa L.) in the wet season (June/July to September/October) and grown under aerobic soil conditions. The remaining fallow period (winter followed by summer) of about 8–9 months leads to natural crash in the population of native arbuscular mycorrhizal fungi (AMF) in the soil. Attempts have been made to minimize this population crash by reducing soil disturbance-induced deleterious effects on native AMF activity of improperly scheduled off-season tillage, an agronomic recommendation for weed and disease (soil-borne) management, practiced by the upland farmers. On-farm (farmers' field) evaluation of effects of all suitable off-season tillage schedule combinations on rice during wet seasons of 2004, 2005, and 2006 revealed that a maximum of two off-season tillage schedules with a minimum gap of 13 weeks between them minimized the population crash of native AMF with a concomitant increase in phosphorus (P) uptake and grain yield of upland rice (variety “Vandana”).     

Isolation and sequence analysis of a β-tubulin gene from arbuscular mycorrhizal fungi

A full-length β-tubulin gene has been cloned and sequenced from Gigaspora gigantea and Glomus clarum, two arbuscular mycorrhizal fungi (AMF) species in the phylum Glomeromyota. The gene in both species is organized into five exons and four introns. Both genes are 94.9% similar and encode a 447 amino acid protein. In comparison with other fungal groups, the amino acid sequence is most similar to that of fungi in the Chytridiomycota. The codon usage of the gene in both AMF species is broad and biased in favor of an A or a T in the third position. The four introns varied in length from 87 to 168 bp for G. gigantea and from 90 to 136 bp for G. clarum. Of all fungi in which full-length sequences have been published, only AMF do not have an intron before codon 174. The introns positioned at codons 174 and 257 in AMF match the position of different introns in β-tubulin genes of some Zygomycete, Basidiomycete, and Ascomycete fungi. The 5′ and 3′ splice site consensus sequences are similar to those found in introns of most fungi. Sequence analysis from single-strand conformation polymorphism analysis confirmed the presence of two β-tubulin gene copies in G. clarum, but only one copy was evident in G. gigantea based on Southern hybridization analysis.     

Arbuscular mycorrhizal mediation of biomass�Cdensity relationship of Medicago sativa L. under two water conditions in a field experiment

The biomass–density relationship (whereby the biomass of individual plants decreases as plant density increases) has generally been explained by competition for resources. Arbuscular mycorrhizal fungi (AMF) are able to affect plant interactions by mediating resource utilization, but whether this AMF-mediated interaction will change the biomass–density relationship is unclear. We conducted an experiment to test the hypothesis that AMF will shift the biomass–density relationship by affecting intraspecific competition. Four population densities (10, 100, 1,000, or 10,000 seedlings per square meter) of Medicago sativa L. were planted in field plots. Water application (1,435 or 327.7 mm/year) simulated precipitation in wet areas (sufficient water) and arid areas (insufficient water). The fungicide benomyl was applied to suppress AMF in some plots (“low-AMF” treatment) and not in others (“high-AMF” treatment). The effect of the AMF treatment on the biomass–density relationship depended on water conditions. High AMF enhanced the decrease of individual biomass with increasing density (the biomass–density line had a steeper slope) when water was sufficient but not when water was insufficient. AMF treatment did not affect plant survival rate or population size but did affect absolute competition intensity (ACI). When water was sufficient, ACI was significantly higher in the high-AMF treatment than in the low-AMF treatment, but ACI was unaffected by AMF treatment when water was insufficient. Our results suggest that AMF status did not impact survival rate and population size but did shift the biomass–density relationship via effects on intraspecific competition. This effect of AMF on the biomass–density relationship depended on the availability of water.     

Taxonomie, écologie et lutte biologique

Summary It is impossible to study modern taxonomy without taking into consideration the population genetic and ecology. Thé Linnean concept of monotypic species should be replaced by the biometric study of the frequency of the variability of the biological characters (particularly morphological) used in taxonomy. The study of populations offers a larger scientific interest than the one of isolated types. It gives the possibility of caracterizing the evolution stability of the species. So, Taxonomy can help considerably in biological control. By revealing large variability species, it gives a criterion of their adjusting plasticity. But, by definition, the biological control is based on the adjusting capacity of the entomophagous insects either to a new host or to a new environment. Information given by the taxonomist on the species stability has consequently a great value in the choice and use of the Entomophagous insects. — Ecologists using biological control, expect from the taxonomist to lie more than a label, which implies the use of biometric analysis methods in Entomophagous taxonomy.      

经典植物分类学的发展与iFlora

简要介绍了经典植物分类的基础工作,即鉴定、描述、命名和分类,以及发展动向。作者认为计算机辅助鉴定（如delta intkey）将成为形态鉴定的常规手段;模式识别技术是植物图像鉴定的发展方向;突出物种遗传本质的分子鉴定将成为植物物种鉴定的核心。DELTA系统或类似的符合计算机逻辑的分类学描述语言将成为新一代植物志（iFlora）的基本语言。近20年来,一些学者逐渐表现出对现行植物命名法规的质疑和修改意愿,特别是生物法规草案（Draft BioCode）和谱系法规（PhyloCode）两个试行法规的诞生。它们促进对法规的革新与完善,并在iFlora中得以体现。以APG系统为代表的分子系统发育研究已经成为植物系统分类研究的主流,但在物种层面,物种的界定仅仅依靠分子信息显然是不够的,而必须运用综合的特征性状信息进行分析解读,真正实现物种在形态、遗传信息等综合性状的融合统一。了解和掌握这些新的技术和研究成果,无疑对于iFlora设计和编研有着重要的参考意义,而基于新技术和新理念的iFlora也将成为植物分类最新发展成果的集中体现。     

Differences of arbuscular mycorrhizal fungal diversity and community between a cultivated land,an old field,and a never-cultivated field in a hot and arid ecosystem of southwest China

We investigated the spore density, species composition, and diversity of arbuscular mycorrhizal fungi (AMF) in a cultivated land (CL), an old field (OF), and a never-cultivated field (NCF), which are located adjacently in a slope in the hot and arid ecosystem of southwest China. AMF spores in the rhizosphere soils of representative plants in the three habitats were extracted by wet-sieving and decanting. A total of 47 taxa of AMF including 31 taxa from the genus Glomus, 8 from Acaulospora, 6 from Scutellospora, 1 from Entrophospora, and 1 from Gigaspora were extracted and identified morphologically. The highest spore density occurred in NCF, slightly lower in OF and lowest in CL, and the Shannon–Wiener index of species diversity was reversed. The dominant species of AMF were different in the three habitats. OF resembled NCF more than CL in AMF spore density, species richness, and community composition, which means that AMF community in the OF has been developing from cultivated land to natural habitat. Cluster analysis based on the similarity in AMF community composition indicated that the distribution of AMF was not random over space and that AMF community composition associated with a given plant species was greatly habitat-convergence. Following the cluster analysis, we hypothesized that the effect of habitats on AMF communities were greater than that of the host preference to AMF. L-F. Li and T. Li contributed equally to this work.     

基因组学时代的真菌分类学:机遇与挑战

真菌物种的形态特征有限,加之形态滞后和形态可塑性,仅靠外部形态、内部结构及生理生化指标,很难把握真菌的系统亲缘。应用DNA测序、基因组测序、比较基因组学及生物信息学等技术,研究人员可以快速识别真菌演化中出现的数量众多的单系支系,为建立各分类等级的新分类单元提供有力证据,为真菌分类学研究带来了新的希望和活力。自2000年以来,在真菌界至少发表了1新亚界、4新门、7新亚门、19新纲、9新亚纲、40余新目等高级分类单元。近3年来,我国发表了20余个真菌新属,其中绝大多数属的建立都有分子证据支持。可以预见,大量的新种、新属、新科乃至更高级分类单元将会在今后10年内持续发现和建立。这必将大大促进真菌分类学的发展,完善现有的真菌分类系统。我们应该顺势而上,利用我国丰富的真菌资源,为真菌分类学的发展做出应有贡献。与此同时,真菌分类学也面临着十分严峻的挑战。挑战主要来自3个方面,一是研究变得越来越综合,不但需要有相应的研究经费支持,而且要求从事该领域的研究人员技术更全面、知识更广博及知识更新速度更快捷;二是新物种描述进度偏慢,远远不能满足人们对物种认识和利用的日益增长的需要;三是研究人员亟需创新研究模式,以新技术、新思路、新机制来构建新的真菌分类学,加速新物种的发现和描述进度,最终为社会进步和科学发展服务。     

Phylogenies from genetic and morphological characters do not support a revision of Gigasporaceae (Glomeromycota) into four families and five genera

The family Gigasporaceae consisted of the two genera Gigaspora and Scutellospora when first erected. In a recent revision of this classification, Scutellospora was divided into three families and four genera based on two main lines of evidence: (1) phylogenetic patterns of coevolving small and large rRNA genes and (2) morphology of spore germination shields. The rRNA trees were assumed to accurately reflect species evolution, and shield characters were selected because they correlated with gene trees. These characters then were used selectively to support gene trees and validate the classification. To test this new classification, a phylogenetic tree was reconstructed from concatenated 25S rRNA and β-tubulin gene sequences using 35% of known species in Gigasporaceae. A tree also was reconstructed from 23 morphological characters represented in 71% of known species. Results from both datasets showed that the revised classification was untenable. The classification also failed to accurately represent sister group relationships amongst higher taxa. Only two clades were fully resolved and congruent among datasets: Gigaspora and Racocetra (a clade consisting of species with spores having one inner germinal wall). Other clades were unresolved, which was attributed in part to undersampling of species. Topology of the morphology-based phylogeny was incongruent with gene evolution. Five shield characters were reduced to three, of which two were phylogenetically uninformative because they were homoplastic. Therefore, most taxa erected in the new classification are rejected. The classification is revised to restore the family Gigasporaceae, within which are the three genera Gigaspora, Racocetra, and Scutellospora. This classification does not reflect strict topology of either gene or morphological evolution. Further revisions must await sampling of additional characters and taxa to better ascertain congruence between datasets and infer a more accurate phylogeny of this important group of fungi.