植物磷营养状况对丛枝菌根真菌生长及代谢活性的调控*

采用四室隔网培养装置,以玉米为宿主植物,通过在植物生长室设0、50、250和500 mgPkg-1 4个施磷水平,研究了植株体内的磷营养状况对AM真菌Glomus sinuosum和Glomus intraradices生长及活性的影响。研究发现在不施磷条件下,接种AM真菌G. intraradices显著促进了植物生长和磷的吸收;低磷条件(50 mgPkg-1)下,接种菌根真菌显著促进了植物对磷的吸收,但对植物生长没有明显的影响;而在高磷条件(250 mgPkg-1 和500 mgPkg-1)下,接种菌根真菌不但没有促进植物的生长和磷的吸收,反而对其有抑制作用。随着施磷水平的提高, AM真菌根内菌丝的碱性磷酸酶活性显著下降;与不施磷相比,低量(50 mgPkg-1)供磷增加了AM真菌土壤中根外菌丝的密度,高磷(250 mgPkg-1 和500 mgPkg-1)降低了土壤中根外菌丝的密度。上述结果说明:⑴ 给宿主植物施用磷肥引起的植物磷营养状况的改变,对AM真菌生长和代谢活性具有一定的调控作用;⑵ G. sinuosum和G. intraradices两种AM真菌的生长和代谢活性对施磷水平的响应程度无显著性差异;⑶ 高磷抑制AM真菌生长和代谢活性,使真菌吸磷量减少,可能是造成菌根效应降低的原因之一     

Branched absorbing structures (BAS): a feature of the extraradical mycelium of symbiotic arbuscular mycorrhizal fungi

The present work describes the morphogenesis and cytological characteristics of 'branched absorbing structures' (BAS, formely named arbuscule-like structures, ALS), small groups of dichotomous hyphae formed by the extraradical mycelium of arbuscular mycorrhizal (AM) fungi. Monoxenic cultures of the AM fungus Glomus intraradices Smith & Schenck and tomato ( Lycopersicum esculentum Mill.) roots allowed the continuous, non-destructive study of BAS development. These structures were not observed in axenic cultures of the fungus under different nutritional conditions or in unsuccessful (asymbiotic) monoxenic cultures. However, extraradical mycelium of G. intraradices formed BAS immediately after fungal penetration of the host root and establishment of the symbiosis. The average BAS development time was 7 d under our culture conditions, after which they degenerated, becoming empty septate structures. Certain BAS were closely associated with spore formation, appearing at the spore's substending hypha. Branches of these spore-associated BAS (spore-BAS) usually formed spores. Electron microscopy studies revealed that BAS and arbuscules show several ultrastructural similarities. The possible role of BAS in nutrient uptake by the mycorrhizal plant is discussed.     

以新鲜根段进行AM真菌的分子检测及竞争性侵染研究

运用nested-PCR技术和AM真菌特异性引物,建立了用新鲜植物根段直接检测AM真菌的分子生物学方法。以真核生物通用引物LR1和NDL22对混合接种的西红柿新鲜根段进行第1次扩增,将其产物进行稀释,再分别以Glomus intraradices 和Glomus mosseae的种特异性引物8.22和5.25进行第2次扩增。在琼脂糖凝胶上观察到AM真菌种特异性条带;运用该技术检测出混合接种时同一根段内不同的AM真菌,并探讨了真菌在植物根部的竞争性侵染。用盆栽方式种植西红柿,混合接种G. intraradices 和G. mosseae,在1个月后,前者侵染占优势。     

Attachment of different soil bacteria to arbuscular mycorrhizal fungal extraradical hyphae is determined by hyphal vitality and fungal species

Attachment of certain bacteria to living arbuscular mycorrhizal fungal extraradical hyphae may be an important prerequisite for interactions between these microorganisms, with implications for nutrient supply and plant health. The attachment of five different strains of gfp-tagged soil bacteria (Paenibacillus brasilensis PB177 (pnf8), Bacillus cereus VA1 (pnf8), Pseudomonas fluorescens SBW25 :: gfp/lux, Arthrobacter chlorophenolicus A6G, and Paenibacillus peoriae BD62 (pnf8)) to vital and nonvital extraradical hyphae of the arbuscular mycorrhizal fungi Glomus sp. MUCL 43205 and Glomus intraradices MUCL 43194 was examined. Arthrobacter chlorophenolicus did not attach to hyphae, whereas the other bacterial strains did to a varying degree. Only P. brasilensis showed greater attachment to vital hyphae than nonvital hyphae of both Glomus species tested. Pseudomonas fluorescens showed a higher attachment to vital compared with nonvital Glomus sp. MUCL 43205 hyphae, whereas this relationship was opposite for attachment to G. intraradices. Both B. cereus and P. peoriae showed higher attachment to nonvital hyphae. This study provides novel evidence that under laboratory conditions soil bacteria differ in their ability to colonize vital and nonvital hyphae and that this can also be influenced by the arbuscular mycorrhizal fungal species involved. The significance of bacterial attachment to mycorrhizal fungal extraradical hyphae is discussed.     

The arbuscular mycorrhizal symbiosis links N mineralization to plant demand

Arbuscular mycorrhizal (AM) fungi facilitate inorganic N (NH₄ ⁺ or NO₃ ⁻) uptake by plants, but their role in N mobilization from organic sources is unclear. We hypothesized that arbuscular mycorrhizae enhance the ability of a plant to use organic residues (ORs) as a source of N. This was tested under controlled glasshouse conditions by burying a patch of OR in soil separated by 20-μm nylon mesh so that only fungal hyphae can pass through it. The fate of the N contained in the OR patch, as influenced by Glomus claroideum, Glomus clarum, or Glomus intraradices over 24 weeks, was determined using ¹⁵N as a tracer. AM fungal species enhanced N mineralization from OR to different levels. N recovery and translocation to Russian wild rye by hyphae reached 25% of mineralized N in G. clarum, which was most effective despite its smaller extraradical development in soil. Mobilization of N by G. clarum relieved plant N deficiency and enhanced plant growth. We show that AM hyphae modify soil functioning by linking plant growth to N mineralization from OR. AM species enhance N mineralization differentially leading to species-specific changes in the quality of the soil environment (soil C-to-N ratio) and structure of the soil microbial community.     

Extraradical hyphae of the mycorrhizal fungus Glomus intraradices can hydrolyse organic phosphate

Organic phosphorus sources make up a large fraction of the total P in some soils. Vesicular–arbuscular mycorrhizal fungi provide a large surface area for the absorption of inorganic P. The question of whether or not they have direct access to organic P by producing extracellular phosphatases has hitherto been controversial because experiments had not been performed in the absence of other soil microorganisms. We used a split-dish in vitro carrot mycorrhiza system free from contaminating microorganisms. The extraradical hyphae of Glomus intraradices hydrolysed both 5-bromo-4-chloro-3-indolyl phosphate and phenolphthalein diphosphate. Moreover, they transferred significantly more P to roots when they had access to inositol hexaphosphoric acid (phytate) than when they did not. Thus we show unequivocally that extraradical hyphae of G. intraradices can hydrolyse organic P, and, further, that the resultant inorganic P can be taken up and transported to host roots.     

Evidence for specificity of cultivable bacteria associated with arbuscular mycorrhizal fungal spores

Bacteria associated with arbuscular mycorrhizal (AM) fungal spores may play functional roles in interactions between AM fungi, plant hosts and defence against plant pathogens. To study AM fungal spore-associated bacteria (AMB) with regard to diversity, source effects (AM fungal species, plant host) and antagonistic properties, we isolated AMB from surface-decontaminated spores of Glomus intraradices and Glomus mosseae extracted from field rhizospheres of Festuca ovina and Leucanthemum vulgare. Analysis of 385 AMB was carried out by fatty acid methyl ester (FAME) profile analysis, and some also identified using 16S rRNA gene sequence analysis. The AMB were tested for capacity to inhibit growth in vitro of Rhizoctonia solani and production of fluorescent siderophores. Half of the AMB isolates could be identified to species (similarity index 0.6) within 16 genera and 36 species. AMB were most abundant in the genera Arthrobacter and Pseudomonas and in a cluster of unidentified isolates related to Stenotrophomonas. The AMB composition was affected by AM fungal species and to some extent by plant species. The occurrence of antagonistic isolates depended on AM fungal species, but not plant host, and originated from G. intraradices spores. AM fungal spores appear to host certain sets of AMB, of which some can contribute to resistance by AM fungi against plant pathogens.     

An arbuscular mycorrhizal inoculum enhances root proliferation in, but not nitrogen capture from, nutrient-rich patches in soil

Most work on root proliferation to a localized nutrient supply has ignored the possible role of mycorrhizal fungi, despite their key role in nutrient acquisition. Interactions between roots of Plantago lanceolata , an added arbuscular mycorrhiza (AM) inoculum and nitrogen capture from an organic patch ( Lolium perenne shoot material) dual-labelled with ¹⁵N and ¹³C were investigated, to determine whether root proliferation and nitrogen (N) capture was affected by the presence of AM fungi. Decomposition of the organic patch in the presence and absence of roots peaked in all treatments at day 3, as shown by the amounts of ¹³CO₂ detected in the soil atmosphere. Plant N concentrations were higher in the treatments with added inoculum 10 d after patch addition, but thereafter did not differ among treatments. Plant phosphorus concentrations at the end of the experiment were depressed by the addition of the organic residue in the absence of mycorrhizal inoculum. Although uninoculated plants were also colonized by mycorrhizal fungi, colonization was enhanced at all times by the added inoculum. Addition of the AM inoculum increased root production, observed in situ by the use of minirhizotron tubes, most pronouncedly within the organic patch zone. Patch N capture by the end of the experiment was c . 7.5% and was not significantly different as a result of adding an AM inoculum. Furthermore, no ¹³C enrichments were detected in the plant material in any of the treatments showing that intact organic compounds were not taken up. Thus, although the added AM fungal inoculum benefited P. lanceolata seedlings in terms of P concentrations of tissues it did not increase total N capture or affect the form in which N was captured by P. lanceolata roots.     

丛枝菌根真菌对植物根尖分生区和根冠细胞的侵染*

一般说来,从枝菌根（ＡＭ）真菌大多数是从植物根系根毛区（成熟区）侵入和扩展的,在显微镜下往往看不到根尖分生区和根冠表皮细胞被ＡＭ真菌侵染的特征。这就很容易给人们造成一种假象,似乎ＡＭ真菌不能侵染根尖分生区和根冠表皮细胞,即它们对ＡＭ真菌是免疫的。然而笔者多次于显微镜下看到ＡＭ真菌侵染根尖分生区和根冠表皮细胞,并形成典型的泡囊、丛枝、菌丝等结构。这一现象导致作者在温室盆栽和大田条件下研究了玫瑰红巨孢囊霉（ Ｇｉｇａｓｐｏｒａ　ｒｏｓｅａ Ｎｉｃｏｌ　＆ Ｓｃｈｅｎｃｋ）、珠状巨孢囊霉（Ｇｉｇａｓｐｏｒａ　ｍａｒｇａｒｉｔａ　Ｂｅｃｋｅｒ　＆　Ｈａｌｌ）、根内球囊霉（Ｇｌｏｍｕｓ　ｏｍｔｒａｒａｄｉｃｅｓ　ｓｃｈｅｎｃｋ　＆　Ｓｍｉｔｈ、摩西球囊霉（Ｇｌｏｍｕｓ ｍｏｓｓｅａｅ （Ｎｉｃｏｌ　＆ Ｇｅｒｄ．） Ｇｅｒｄｅｍａｎｎ　＆ Ｔｒａｐｐｅ）、地表球囊霉（ Ｇｌｏｍｕｓ ｖｅｒｓｉｆｏｒｍｅ（ Ｋａｒｓｔｅｎ）Ｂｅｒｃｈ）和弯丝硬囊霉（ Ｓｃｌｅｒｏｃｙｓｔｉｓ　ｓｉｎｕｏｓａ　Ｇｅｒｄｅｍａｎｎ　＆ Ｂａｋｈｉ）对棉花（Ｇｏｓｓｙｐｉｕｍ　ｈｉｒｓｕｔｕｍ　Ｌ．）、烟草（Ｎｉｃｏｔｉａｎａ　 ｔａｂａｃｕｍ Ｌ．）和白     

Absence of carbon transfer between Medicago truncatula plants linked by a mycorrhizal network, demonstrated in an experimental microcosm

Carbon transfer between plants via a common extraradical network of arbuscular mycorrhizal (AM) fungal hyphae has been investigated abundantly, but the results remain equivocal. We studied the transfer of carbon through this fungal network, from a Medicago truncatula donor plant to a receiver (1) M. truncatula plant growing under decreased light conditions and (2) M. truncatula seedling. Autotrophic plants were grown in bicompartmented Petri plates, with their root systems physically separated, but linked by the extraradical network of Glomus intraradices. A control Myc-/Nod- M. truncatula plant was inserted in the same compartment as the receiver plant. Following labeling of the donor plant with 13CO2, 13C was recovered in the donor plant shoots and roots, in the extraradical mycelium and in the receiver plant roots. Fatty acid analysis of the receiver's roots further demonstrated 13C enrichment in the fungal-specific lipids, while almost no label was detected in the plant-specific compounds. We conclude that carbon was transferred from the donor to the receiver plant via the AM fungal network, but that the transferred carbon remained within the intraradical AM fungal structures of the receiver's root and was not transferred to the receiver's plant tissues.     

Mycorrhizal mediation of plant N acquisition and residue decomposition: Impact of mineral N inputs

Mycorrhizas are ubiquitous plant–fungus mutualists in terrestrial ecosystems and play important roles in plant resource capture and nutrient cycling. Sporadic evidence suggests that anthropogenic nitrogen (N) input may impact the development and the functioning of arbuscular mycorrhizal (AM) fungi, potentially altering host plant growth and soil carbon (C) dynamics. In this study, we examined how mineral N inputs affected mycorrhizal mediation of plant N acquisition and residue decomposition in a microcosm system. Each microcosm unit was separated into HOST and TEST compartments by a replaceable mesh screen that either prevented or allowed AM fungal hyphae but not plant roots to grow into the TEST compartments. Wild oat (Avena fatua L.) was planted in the HOST compartments that had been inoculated with either a single species of AM fungus, Glomus etunicatum, or a mixture of AM fungi including G. etunicatum. Mycorrhizal contributions to plant N acquisition and residue decomposition were directly assessed by introducing a mineral ¹⁵N tracer and ¹³C‐rich residues of a C₄ plant to the TEST compartments. Results from ¹⁵N tracer measurements showed that AM fungal hyphae directly transported N from the TEST soil to the host plant. Compared with the control with no penetration of AM fungal hyphae, AM hyphal penetration led to a 125% increase in biomass ¹⁵N of host plants and a 20% reduction in extractable inorganic N in the TEST soil. Mineral N inputs to the HOST compartments (equivalent to 5.0 g N m^?2 yr^?1) increased oat biomass and total root length colonized by mycorrhizal fungi by 189% and 285%, respectively, as compared with the no‐N control. Mineral N inputs to the HOST plants also reduced extractable inorganic N and particulate residue C proportion by 58% and 12%, respectively, in the corresponding TEST soils as compared to the no‐N control, by stimulating AM fungal growth and activities. The species mixture of mycorrhizal fungi was more effective in facilitating N transport and residue decomposition than the single AM species. These findings indicate that low‐level mineral N inputs may significantly enhance nutrient cycling and plant resource capture in terrestrial ecosystems via stimulation of root growth, mycorrhizal functioning, and residue decomposition. The long‐term effects of these observed alterations on soil C dynamics remain to be investigated.     

野生植物根围的丛枝菌根真菌Ⅱ

本文主要报道了野生植物根围Glomus属的17个种,聚球囊霉G.aggregatumSchenck＆Smith,苏格兰球囊霉G.caledonium（Nicol.＆Gerd.）Trappe＆Gerd,近明球囊霉G．claroideumSchenck＆Smith,明球囊霉G．clarumNicolson＆Schenck,缩球囊霉G．constrictumTrappe,透光球囊霉G．diaphanumMorton,幼套球囊霉G．etunicatumBecker＆Gerdemann,集球囊霉G．fasciculatum（Thaxter）Gerd．＆Trappe,何氏球囊霉G．hoiBerch＆Trappe,地球囊霉G.geosporum（Nicol．＆Derd.）Warker,根内球囊霉G．intraradicesSchenck＆Smith,摩西球囊霉G．mosseae（Nicol．＆Gerd.）Gerd.＆Trappe,隐球囊霉G.occultumWalker,网状球囊霉G.reticulatumBhattcharjee＆Mukerji,地表球囊霉G.versiforme（Karsten）Berch,台湾球囊霉G．formosanumWu＆Chen,悬钩子球囊霉G．rubiformeGerdemann＆Trappe）Almeida＆Schenck;内养囊霉属1个种,稀有内养囊霉Entrophosporainfrequens（Hall）Ames＆Schenider。其中,网状球囊霉为我国新记录种。     

Tracking carbon from the atmosphere to the rhizosphere

Turnover rates of arbuscular mycorrhizal (AM) fungi may influence storage of soil organic carbon (SOC). We examined the longevity of AM hyphae in monoxenic cultures; and we also used ¹³C incorporation into signature fatty acids to study C dynamics in a mycorrhizal symbiosis involving Glomus intraradices and Plantago lanceolata. ¹³C enrichment of signature fatty acids showed rapid transfer of plant assimilates to AM fungi and a gradual release of C from roots to rhizosphere bacteria, but at a much slower rate. Furthermore, most C assimilated by AM fungi remained 32 days after labelling. These findings indicate that ¹³C labelled fatty acids can be used to track C flux from the atmosphere to the rhizosphere and that retention of C in AM fungal mycelium may contribute significantly to SOC.     

植酸钠对菌根真菌根内菌丝碱性磷酸酶活性及根外菌丝生长的影响

采用三室隔网培养装置,以玉米为宿主植物,接种丛枝菌根真菌(AM)(Glomus intraradices),研究了不同用量的植酸钠对AM真菌生长和代谢活性的影响．研究发现,接种AM真菌的植株地上部和根系的P浓度和吸P量,比非菌根植物的提高了1～2倍．外源植酸钠的存在,显著降低了AM真菌根内菌丝的碱性磷酸酶活性,增加了AM真菌在土壤中的菌丝密度．结果表明,外源植酸钠对根内AM真菌碱性磷酸酶活性和真菌根外菌丝的生长具有调控(增减)作用,并且AM真菌提高了植物对土壤固有养分和外源植酸钠中P的吸收和利用．     

Expressed genes in the extraradical hyphae of an arbuscular mycorrhizal fungus, Glomus intraradices, in the symbiotic phase

To collect extraradical hyphae of arbuscular mycorrhizal (AM) fungi for RNA isolation, a PVDF membrane was laid on the hyphal compartment of a two-compartment culture system of transformed carrot hairy roots and Glomus intraradices. Extraradical hyphae free from host tissue were easily collected, and their RNA was rapidly extracted with a modified acid guanidinium thiocyanate-phenol-chloroform method. A 3'-RACE (rapid amplification of cDNA ends) of a known gene indicated that this protocol enabled the isolation of mRNA molecules as small as 2.3 kb. The cDNA libraries of an AM fungus from the aseptic extraradical hyphae in a symbiotic state were constructed for the first time. Three-fourth of 150 ESTs (expressed sequence tags) indicated low or no similarities to known sequences from other organisms.