新疆北部地区常见植物根围的丛枝菌根真菌

从新疆北部地区24种常见植物根围土壤中分离并鉴定出5属27种丛枝菌根真菌,其中球囊霉属Glomus15种,无梗囊霉属Acaulospora9种,原囊霉属Archaeospora1种,内养囊霉属Entrophospora1种,巨孢囊霉属Gigaspora1种。道氏球囊霉Glomusdominikii是新记录种。     

福建漳州常见药用植物根围的丛枝菌根真菌

为了解闽南地区药用植物根围丛枝菌根真菌多样性分布,作者调查了福建省漳州市20种常见药用植物根围的AM真菌。从福建省漳州市小溪镇、国强乡等地共分离出无梗囊霉属Acaulospora 12种、原囊霉属Archaeospora 1种、巨孢囊霉属Gigaspora 2种、球囊霉属Glomus 42种、盾巨孢囊霉属Scutellospora 9种,其中沙生球囊霉Glomus arenarium、金黄球囊霉Glomus aureum和厚皮球囊霉Glomus callosum等3种为我国新记录种。     

Assessing the diversity of AM fungi in arid gypsophilous plant communities

In the present study, we used PCR-Single-Stranded Conformation Polymorphism (SSCP) techniques to analyse arbuscular mycorrhizal fungi (AMF) communities in four sites within a 10 km² gypsum area in Southern Spain. Four common plant species from these ecosystems were selected. The AM fungal small-subunit (SSU) rRNA genes were subjected to PCR, cloning, SSCP analysis, sequencing and phylogenetic analyses. A total of 1443 SSU rRNA sequences were analysed, for 21 AM fungal types: 19 belonged to the genus Glomus , 1 to the genus Diversispora and 1 to the Scutellospora. Four sequence groups were identified, which showed high similarity to sequences of known glomalean species or isolates: Glo G18 to Glomus constrictum , Glo G1 to Glomus intraradices , Glo G16 to Glomus clarum , Scut to Scutellospora dipurpurescens and Div to one new genus in the family Diversisporaceae identified recently as Otospora bareai . There were three sequence groups that received strong support in the phylogenetic analysis, and did not seem to be related to any sequences of AM fungi in culture or previously found in the database; thus, they could be novel taxa within the genus Glomus : Glo G4, Glo G2 and Glo G14. We have detected the presence of both generalist and potential specialist AMF in gypsum ecosystems. The AMF communities were different in the plant studied suggesting some degree of preference in the interactions between these symbionts.     

Geosiphon pyriforme,a fungus forming endocytobiosis withNostoc (Cyanobacteria), is an ancestral member of the glomales: Evidence by SSU rRNA Analysis

Geosiphon pyriforme inhabiting the surface of humid soils represents the only known example of endocytobiosis between a fungus (Zygomycotina; macrosymbiont) and cyanobacteria (Nostoc; endosymbiont). In order to elucidate the taxonomical and evolutionary relationship ofGeosiphon pyriforme to fungi forming arbuscular mycorrhiza (AM fungi), the small-subunit (SSU) ribosomal RNA genes ofGeosiphon pyriforme andGlomus versiforme (Glomales; a typical AM fungus) were analyzed and aligned with SSU rRNA sequences of several Basidiomycetes, Ascomycetes, Chytridiomycetes, and Zygomycetes, together with all AM-fungal (Glomales) sequences published yet. The distinct group of the order Glomales, which includesGeosiphon, does not form a clade with any other group of Zygomycetes. Within the Glomales, two main lineages exist. One includes the families Gigasporaceae and Acaulosporaceae; the other one is represented by the genusGlomus, the members of which are very divergent.Glomus etunicatum andGeosiphon pyriforme both form independent lineages ancestral to the Glomales. The data provided by the present paper confirm clearly thatGeosiphon represents a fungus belonging to the Glomales. The question remains still open as to whether or notGeosiphon is to be placed within or outside the genusGlomus, since this genus is probably polyphyletic and not well defined yet.Geosiphon shows the ability of aGlomus-like fungus to form a “primitive” symbiosis with a unicellular photcautotrophic organism, in this case a cyanobacterium, leading to the conclusion that a hypothetical association of aGlomus-like fungus with a green alga as a step during the evolution of the land plants appears probable. Correspondence to: H. Gehrig     

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.     

Glomales SSUrRNA gene diversity

The arbuscular mycorrhiza (AM) is one of the most important symbioses on earth, formed between about 80% of vascular plants and AM fungi (Smith & Read, 1997), which are placed in the order Glomales (Morton & Benny, 1990). A discussion has recently arisen about the rDNA-gene and general genome organization of this fungal group, which, as far as is known, does not possess sexuality (Sanders, 1999). This discussion mainly arose as a result of the assumption that one spore of an AM fungus ( Scutellospora castanea ) contains small subunit (SSU) rRNA genes that belong, phylogenetically, to different families within the Glomales, namely the Gigasporaceae and Glomaceae (Hosny et al ., 1999). Questions about genetic drift, infrequent recombination, concerted evolution and the validity of phylogenetic techniques have arisen (Hosny et al ., 1999; Sanders, 1999). However, this assumption might well be based on the sequence of an Ascomycete contaminant.     

Comparison of morphological and molecular genetic quantification of relative abundance of arbuscular mycorrhizal fungi within roots

Nested PCR amplicons of ribosomal RNA genes have been used to identify individuals within assemblages of arbuscular mycorrhizal (AM) fungi in roots and to estimate their relative abundance. Microscopy has also been used to identify their relative abundance in roots, but only at low resolution, usually the genus level. We evaluated the robustness of using nested PCR amplicons of ribosomal RNA genes to estimate the relative abundance of undefined AM fungi in uniformly aged roots in comparison to visual estimates. The relative abundance of AM fungi was assessed as per cent root length colonised by morphotypes and relative sequence type abundance in clone libraries. Plants were grown in coastal soil to obtain assemblages of unknown AM fungi at two times (spring and autumn). Relative abundance of dominant genera of AM fungi in roots (Archaeospora and Glomus) based on an analysis of ribosomal RNA genes did not consistently correspond with relative abundance of morphotypes. This microscopic vs. molecular genetic comparison supports previous conclusions that there can be limitations in using nested PCR amplicons for quantifying the relative abundance of AM fungi in roots, with a sampling bias likely to be of significance. Both molecular genetic and morphological methods are used to estimate relative abundance of AM fungi as a precursor to understanding mycorrhizal function in field soils, but they are rarely verified using alternative approaches although this may be necessary.     

都江堰亚热带地区常见植物根围的丛枝菌根真菌*

从都江堰亚热带地区常见植物根围土壤中分离并鉴定了4属27种丛枝菌根真菌,其中无柄囊霉属Acaulospora4种,原囊霉属Archaeospora1种,内养囊霉属Entrophospora2种,球囊霉Glomus20种。其中哥伦比亚内养囊霉Entrophosporacolombiana,澳洲球囊霉Glomusaustrale,卷曲球囊霉G.convolutum,肿涨球囊霉G.gibbosum,球泡球囊霉G.globiferum,异形球囊霉G.heterosporum(A)为我国六个新记录种。     

Molecular diversity of arbuscular mycorrhizal fungi colonising Hyacinthoides non-scripta (bluebell) in a seminatural woodland

Arbuscular mycorrhizal (AM) fungi form symbiotic associations with plant roots. Around 150 species have been described and it is becoming clear that many of these species have different functional properties. The species diversity of AM fungi actively growing in roots is therefore an important component of ecosystem diversity. However, it is difficult to identify AM fungi below the genus level from morphology in planta , as they possess few informative characters. We present here a molecular method for identifying infrageneric sequence types that estimate the taxonomic diversity of AM fungi present in actively growing roots. Bluebell roots were sampled from beneath two different canopy types, oak and sycamore, and DNA sequences were amplified from roots by the polymerase chain reaction with fungal-specific primers for part of the small subunit ribosomal RNA gene. Restriction fragment length polymorphism among 141 clones was assessed and 62 clones were sequenced. When aligned, discrete sequence groups emerged that cluster into the three families of AM fungi: Acaulosporaceae, Gigasporaceae and Glomaceae. The sequence variation is consistent with rRNA secondary structure. The same sequence types were found at both sampling times. Frequencies of Scutellospora increased in December, and Acaulospora increased in abundance in July. Sites with a sycamore canopy show a reduced abundance of Acaulospora , and those with oak showed a reduced abundance of Glomus . These distribution patterns are consistent with previous morphological studies carried out in this woodland. The molecular method provides an alternative method of estimating the distribution and abundance of AM fungi, and has the potential to provide greater resolution at the infrageneric level.     

Identification of arbuscular mycorrhizal fungi in soils and roots of plants colonizing zinc wastes in southern Poland

 Analysis of the community of arbuscular mycorrhizal (AM) fungi in roots of Fragaria vesca growing in a heavy metal contaminated site was carried out on a Zn waste site near Chrzanow (southern Poland). The waste substratum was characterized by high contents of Pb, Zn, Cd, Cu and As, and by low levels of N, P and organic matter. Spores of Glomales were isolated by wet sieving and DNA was isolated from individual spores. Nested polymerase chain reaction (PCR) with taxon-specific primers was used to identify the species Glomus mosseae, Glomus intraradices and Glomus claroideum. Spores of other fungi were morphologically characterized and new taxon-discriminating molecular probes were developed for two of them (Glomus sp. HM-CL4 and HM-CL5) based on variations in the large ribosomal subunit (25S rDNA). High sequence similarities were found between Glomus sp. HM-CL4 and Glomus gerdemanii, and between Glomus sp. HM-CL5 and Glomus occultum. The designed primers were used to characterize the population of AM fungi colonizing the roots of F. vesca collected from the Zn waste site. The analysis, carried out on roots stained with trypan blue, showed that the most effective colonizer was closely related to G. gerdemannii. G. claroideum and the G. occultum-like fungus were slightly less common whilst frequencies of G. intraradices and G. mosseae in roots were much lower. The analysis of mycorrhiza stained with rhodizoniate to localize heavy metal accumulation showed that the stain does not influence the PCR reaction. Seventy percent of the root samples containing positively stained fungal hyphae were found to be colonized by G. mosseae. The data obtained demonstrate the usefulness of nested PCR for studies carried out in polluted areas. It will enable selection of AM fungi which are able to colonize plant roots under heavy metal stress conditions, as well as the identification of fungi showing high in situ accumulation of potentially toxic elements. Accepted: 7 July 2000     

Fungal spore diversity of arbuscular mycorrhizal fungi associated with spring wheat: effects of tillage

We investigated the influence of tilling, N fertilization and crop stage on arbuscular mycorrhizae (AM) fungal species diversity in a wheat monoculture in the Pampa region of Argentina. Glomalean spores were isolated by wet sieving and decanting from conventionally tilled and nontilled soils cropped with wheat with or without N fertilization, at three phenological stages of the crop (tilling, flowering and grain filling) and fallow. Morphological characterization yielded at least 24 AM fungi taxa in the field samples, belonging to six genera of AMF: Acaulospora Archaeospora, Entrophospora, Gigaspora, Glomus and Scutellospora. Tilling and fertilization treatments did not result in decreased spore biodiversity. Wheat phenology influenced AM communities, with highest spore biodiversity during grain filling.     

Molecular analysis of Gigaspora (Glomales, Gigasporaceae)

This work presents a cooperative effort to integrate new molecular (isozyme and SSU analyses) characters into the morphological taxonomy of the genus Gigaspora (Glomales). Previous analyses of published Gigaspora SSU sequences indicated the presence of a few polymorphic nucleotides in the region delimited by primers NS71-SSU 1492'. In our study, the SSU of 24 isolates of arbuscular mycorrhizal (AM) fungi from the Gigasporaceae were amplified and the NS71-SSU 1492' region was directly sequenced. The corresponding sequences of four more isolates of AM fungi from Gigasporaceae, already published, were also included in our analyses. Three Gigaspora groups were identified on the basis of a 6 nucleotide-long 'molecular signature': Gigaspora rosea group ( G. rosea + G. albida ), Gigaspora margarita group ( G. margarita + G. decipiens ) and Gigaspora gigantea , which constituted a group by itself. The isozyme profiles (malate dehydrogenase, MDH) of 12 of these 28 isolates, and seven other isolates not sequenced, were compared. The results obtained further supported the grouping of isolates provided by the SSU analysis. Both SSU and MDH analysis indicated that two out of the 35 isolates had been misidentified, which was confirmed when their morphology was reassessed. The use of the Gigaspora intrageneric molecular signature as a quick, unambiguous and objective method to recognize Gigaspora isolates under any (field or laboratory) experimental conditions is suggested.     

金沙江干热河谷(元谋段)丛枝菌根真菌多样性研究

从金沙江干热河谷(元谋段)75种植物的根际土壤中分离鉴定了44种丛枝菌根真菌, 分属无梗囊霉属Acaulospora、古孢霉属Archaeospora、内养囊霉属Entrophospora、巨孢囊霉属Gigaspora、球囊霉属Glomus和盾巨孢囊霉属Scutellospora, 其中,球囊霉属和无梗囊霉属为金沙江干热河谷中丛枝菌根真菌的优势属。齿状无梗囊霉A. denticulata、刺状无梗囊霉A. spinosa、瘤状无梗囊A. tuberculata,近明球囊霉Glomus claroideum、明球囊霉G. clarum、根内球囊霉G. intraradices、单孢球囊霉G. monosporum、弯丝球囊霉G. sinuosa是金沙江干热河谷(元谋段)的优势种。金沙江干热河谷土壤中丛枝菌根真菌的孢子密度为5~6400个/100g土壤,平均1504;每个根际土壤中丛枝菌根真菌的物种丰富度1~18种,平均9种。     

金沙江干热河谷(元谋段)丛枝菌根真菌多样性研究*

从金沙江干热河谷(元谋段)75种植物的根际土壤中分离鉴定了44种丛枝菌根真菌, 分属无梗囊霉属Acaulospora、古孢霉属Archaeospora、内养囊霉属Entrophospora、巨孢囊霉属Gigaspora、球囊霉属Glomus和盾巨孢囊霉属Scutellospora, 其中,球囊霉属和无梗囊霉属为金沙江干热河谷中丛枝菌根真菌的优势属。齿状无梗囊霉A. denticulata、刺状无梗囊霉A. spinosa、瘤状无梗囊A. tuberculata,近明球囊霉Glomus claroideum、明球囊霉G. clarum、根内球囊霉G. intraradices、单孢球囊霉G. monosporum、弯丝球囊霉G. sinuosa是金沙江干热河谷(元谋段)的优势种。金沙江干热河谷土壤中丛枝菌根真菌的孢子密度为5~6400个/100g土壤,平均1504;每个根际土壤中丛枝菌根真菌的物种丰富度1~18种,平均9种。     

中国盐碱土壤中AM菌的生态分布

对我国盐碱土壤中丛枝菌根（Ａｒｂｕｓｃｕｌａｒ Ｍｙｃｏｒｒｈｉｚａ,ＡＭ） 菌的种属构成、生态分布状况进行了研究．结果表明,不同地区ＡＭ 菌种属构成不同,其种属组成、分布与土壤类型、碱化度和土壤有机质含量有关．盐渍化砂土、壤土和粘土中,Ｇｌｏｍｕｓ 属的真菌数量最多,Ａｃａｕｌｏｓｐｏｒａ 属次之,而Ｇｌｏｍｕｓ 属中的Ｇ．ｍｏｓｓｅａｅ 则是分布最为广泛的菌种．随土壤碱化度的增加,Ｇｌｏｍｕｓ ｍｏｓｓｅａｅ 出现频率随之相对增加．在一定范围内有机质含量越高,土壤中ＡＭ 菌种和属的种类就越多．ＡＭ 菌的种属组成因不同寄主植物而异,其中豆科植物根围中ＡＭ 菌分布的种属数量最多．     

Host-related variability in arbuscular mycorrhizal fungal structures in roots of <Emphasis Type="Italic">Hedera rhombea</Emphasis>, <Emphasis Type="Italic">Rubus parvifolius</Emphasis>, and <Emphasis Type="Italic">Rosa multiflora</Emphasis> under controlled conditions

The arbuscular mycorrhizal (AM) morphology of three host plant species inoculated with single and mixed fungal culture and the distribution of AM fungal species in roots of the hosts treated with a mixed culture of AM fungi were determined. The aim was to investigate the effect of host plants and AM fungi on AM morphology of coexisting plant species. Noncolonized rooted cuttings of Hedera rhombea (Miq) Bean, Rubus parvifolius L., and Rosa multiflora Thunb. were inoculated with five fungal species as single and mixed culture inocula. The fungal species used were Gigaspora rosea and Scutellospora erythropa, previously isolated from H. rhombea; Acaulospora longula and Glomus etunicatum from R. parvifolius; and Glomus claroideum from both plant species. A few hyphal and arbusculate coils were seen in the mixed culture-inoculated roots of R. parvifolius; all fungal treatments produced this Paris-type AM in H. rhombea and Arum-type AM in R. parvifolius, and R. multiflora indicates that AM morphology is strongly controlled by the identity of the host plants used in this study. AM fungal rDNA was extracted separately from roots of each replicate plant species inoculated with the mixed fungal culture, amplified, cloned, sequenced, and analyzed to determine the AM fungal species and their respective proportions in roots of each plant species. Glomus etunicatum and G. claroideum of the family Glomaceae generally occurred more frequently in R. parvifolius and R. multiflora, which form Arum-types, whereas S. erythropa, of the family Gigasporaceae, was the most frequently detected species in H. rhombea, which produced Paris-type AM. Although the genotype of the plant species used appears to determine the AM morphologies formed, there was preferential association between the hosts and AM fungal inoculants.     

A first phylogenetic analysis of Giant Pill-Millipedes (Diplopoda: Sphaerotheriida), a new model Gondwanan taxon, with special emphasis on island gigantism

The phylogeny of the Giant Pill-Millipedes, order Sphaerotheriida, is investigated using a new morphological character matrix comprising 89 characters. The majority of these characters are employed for the first time in millipedes. All trees obtained agree on the monophyletic status of the Sphaerotheriida and several of its tribes, each restricted to a modern land mass. The species from Madagascar displaying island gigantism do not form a monophyletic group. The classic division of Giant Pill-Millipedes into two families, Sphaerotheriidae and Zephronidae, was not reflected in the analysis. The genus Procyliosoma is the sister-group to all other Sphaerotheriida, rendering the family Sphaerotheriidae paraphyletic. A new family-level classification of Giant Pill-Millipedes, based on the current phylogeny, is introduced. The new family Procyliosomatidae contains only the genus Procyliosoma , distributed in Australia and New Zealand. The family Zephronidae remains unchanged, while the family Sphaerotheriidae now incorporates only the African Giant Pill-Millipede genera. All genera from southern India and Madagascar form a monophyletic group and are placed in the new family Arthrosphaeridae. The Malagasy genus Sphaeromimus is more closely related to the Indian Arthrosphaera species than to other genera from Madagascar. A biogeographical analysis identifies the group as a Gondwana taxon (with a notable absence from South America). The current phylogeny of Giant Pill-Millipede families mirrors perfectly the suggested break-up of Gondwana fragments 160–90 Ma. No evidence for a dispersal event could be found, highlighting the importance of Giant Pill-Millipedes as a potential model taxon.     

五指山常见热带树种的丛枝菌根真菌多样性

采用野外调查的方法,分析了五指山不同海拔高度7个科10种常见热带树种形成丛枝菌根(Arbuscular Mycorrhizal,AM)的状况及其根际土壤中AM真菌的多样性。结果表明,所调查的10种热带常见树种都能形成AM共生体,其菌根侵染率随寄主植物的不同,从21.8%～90.5%变化不等,同时,在10种常见植物的根系中也都观察到了AM真菌的典型结构——丛枝和泡囊。从10种植物的根际土壤中共分离到36种AM真菌,隶属于Acaulospora,Glomus,Gigaspora和Scutellospora4个属,其中,Glomus属的真菌是该地区的优势类群,其出现频度和相对多度分别为84%和56%。在所调查的10种热带常见树种中,Swietenia macrophylla根际AM真菌的孢子最丰富,密度高达7.32;Machilus namu根际的AM真菌种类则最为丰富,多样性指数达到1.6548。通过对不同海拔高度Swietenia macrophylla根际AM真菌分布的分析表明,海拔高度显著影响着AM真菌的分布,Gigaspora属的真菌随海拔高度的增加显著升高,Scutellospora属的真菌则显著降低。     

The liverwort Marchantia foliacea forms a specialized symbiosis with arbuscular mycorrhizal fungi in the genus Glomus

Microscopic evidence suggests that fungi forming endosymbioses with liverworts in the Marchantiales are arbuscular mycorrhizal (AM) fungi from the Glomeromycota. Polymerase chain reaction amplification of ribosomal sequences confirmed that endophytes of the New Zealand liverwort, Marchantia foliacea, were members of the genus Glomus. Endophytes from two Glomus rDNA phylotypes were repeatedly isolated from geographically separated liverwort samples. Multiple phylotypes were present in the same liverwort patch. The colonizing Glomus species exhibited substantial internal transcribed spacer sequence variation within phylotypes. This work suggests that certain liverwort species may serve as a model for studying DNA sequence variation in colonizing AM phylotypes and specificity in AM-host relationships.     

Overlapping expression patterns and differential transcript levels of phosphate transporter genes in arbuscular mycorrhizal,P<Subscript>i</Subscript>-fertilised and phytohormone-treated <Emphasis Type="Italic">Medicago truncatula</Emphasis> roots

A microarray carrying 5,648 probes of Medicago truncatula root-expressed genes was screened in order to identify those that are specifically regulated by the arbuscular mycorrhizal (AM) fungus Gigaspora rosea, by P_i fertilisation or by the phytohormones abscisic acid and jasmonic acid. Amongst the identified genes, 21% showed a common induction and 31% a common repression between roots fertilised with P_i or inoculated with the AM fungus G. rosea, while there was no obvious overlap in the expression patterns between mycorrhizal and phytohormone-treated roots. Expression patterns were further studied by comparing the results with published data obtained from roots colonised by the AM fungi Glomus mosseae and Glomus intraradices, but only very few genes were identified as being commonly regulated by all three AM fungi. Analysis of P_i concentrations in plants colonised by either of the three AM fungi revealed that this could be due to the higher P_i levels in plants inoculated by G. rosea compared with the other two fungi, explaining that numerous genes are commonly regulated by the interaction with G. rosea and by phosphate. Differential gene expression in roots inoculated with the three AM fungi was further studied by expression analyses of six genes from the phosphate transporter gene family in M. truncatula. While MtPT4 was induced by all three fungi, the other five genes showed different degrees of repression mirroring the functional differences in phosphate nutrition by G. rosea, G. mosseae and G. intraradices. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.