The role of in vitro cultivation on symbiotic trait and function variation in a single species of arbuscular mycorrhizal fungus

In vitro propagation of AM fungi using transformed root cultures (TRC) is commonly used to obtain pure AM fungal propagules for use in research and industry. Early observations indicate that such an artificial environment can alter traits and function of AM fungi over time. We hypothesized that increased in vitro cultivation may promote ruderal strategies in fungi by enhancing propagule production and reducing mutualistic quality. To examine the effect of in vitro cultivation on the trait and function of AM fungi, we inoculated plants with 11 Rhizoglomus irregulare isolates which fell along a cultivation gradient spanning 80 generations. We harvested plants at 10, 20 and 30 d post inoculation to observe differences in fungal and plant traits post infection. In vitro cultivation led to increased spore production but reduced plant shoot phosphorus. Our results indicate that in vitro propagation may indirectly select for traits that affect symbiotic quality.     

Arbuscular mycorrhizal fungi protect a subtropical tree species exposed to simulated acid rain by accelerating photosynthetic ability,antioxidant enzymes and osmolyte accumulation

丛枝菌根真菌对其宿主光合能力、抗氧化酶和渗透物质积累的促进作用 及其抗酸雨机制的探讨 酸雨在中国南方发生频繁,对亚热带树种生长具有明显抑制作用。以往研究表明,丛枝菌根真菌(AM真菌)可以缓解酸雨对宿主植物的胁迫效应。榉树(Zelkova serrata)为中国南方主要经济树种之一,其如何与共生AM真菌协同、增强其抗酸雨胁迫的能力是本项研究所要探讨的关键科学问题。通过温室控制实验,将榉树幼苗随机接受4个水平的AM真菌接种处理(接种灭菌菌种;单独接种Rhizophagus intraradices;单独接种Diversispora versiformis;接种这两种菌种的混合菌种)和3个pH水平(pH2.5、pH4.0和pH5.6)的硫酸型酸雨和硝酸型酸雨处理组成的12个处理组合,同时测定其生长、光合性能、抗氧化酶、渗透调节和土壤酶的响应格局。研究发现酸雨处理显著降低了非菌根榉树幼苗的总干重、总叶绿素含量、叶片净光合速率和可溶性蛋白的含量;接种AM真菌,特别是接种混合菌种,显著提高了强酸胁迫下榉树幼苗的总干重、光合性能、丙二醛、过氧化物酶、超氧化物歧化酶、可溶性蛋白和根系酸性磷酸酶活性。此外,菌根效应依赖于AM真菌的种类和酸胁迫的梯度。本研究 结果表明,AM真菌对榉树幼苗抗酸胁迫的调控作用主要源于调节宿主植株光合能力、抗氧化酶和渗透物质的积累。榉树与其共生AM真菌在应对酸胁迫上协同机制的解析为该树种在中国南方酸雨区的栽培提供理论基础、具有重要的实践指导意义。     

丛枝菌根网络介导的植物间信号交流研究进展及展望

丛枝菌根(arbuscularmycorrhizal,AM)真菌是一类能够与绝大多数陆地植物形成共生关系的土壤真菌,其根外菌丝可以侵染不同植物根系且可以进行菌丝融合,从而形成丛枝菌根网络(arbuscular mycorrhizal networks, AMNs)。AMNs可以在植物之间转运水分及营养元素如碳(C)、氮(N)、磷(P)等,最近研究表明AMNs还可以在植物遭受环境胁迫时向邻近植物传递防御信号,对周围植物起到“预警”作用。目前,关于环境胁迫条件下AMNs介导的信号物质传递研究仍处于起步阶段,许多问题亟待回答。该文首先回顾了目前有关AMNs介导的信号物质传递研究进展,继而梳理了这一研究领域值得进一步探究的科学问题,包括AMNs在植物间传递防御信号的可能途径及相关机制, AMNs介导的信号传递对菌根共生体系的可能影响,以及AMNs研究中常用的技术及其发展,最后讨论了AMNs介导的信号物质传递在作物保护等方面的可能应用。     

Community structure of ectomycorrhizal fungi in a Pinus muricata forest: minimal overlap between the mature forest and resistant propagule communities

We have investigated colonization strategies by comparing the abundance and frequency of ectomycorrhizal fungal species on roots in a mature Pinus muricata forest with those present as resistant propagules colonizing potted seedlings grown in the same soil samples. Thirty-seven fungal species were distinguished by internal transcribed spacer (ITS) restriction fragment length polymorphisms (RFLPs); most were identified to species level by sporocarp RFLP matches or to genus/family level by using sequence databases for the mitochondrial and nuclear large-subunit rRNA genes. The below-ground fungal community found in the mature forest contrasted markedly with the resistant propagule community, as only four species were found in both communities. The dominant species in the mature forest were members of the Russulaceae, Thelephorales and Amanitaceae. In contrast, the resistant propagule community was dominated by Rhizopogon species and by species of the Ascomycota. Only one species, Tomentella sublilacina (Thelephorales), was common in both communities. The spatial distribution of mycorrhizae on mature roots and propagules in the soil differed among the dominant species. For example, T. sublilacina mycorrhizae exhibited a unique bias toward the organic horizons, Russula brevipes mycorrhizae were denser and more clumped than those of other species and Cenococcum propagules were localized, whereas R. subcaerulescens propagules were evenly distributed. We suggest that species differences in resource preferences and colonization strategies, such as those documented here, contribute to the maintenance of species richness in the ectomycorrhizal community.     

Diversity of arbuscular mycorrhizal fungi colonising roots of the grass species<Emphasis Type="Italic"> Agrostis capillaris</Emphasis> and<Emphasis Type="Italic"> Lolium perenne</Emphasis> in a field experiment

Analysis of arbuscular mycorrhizal (AM) fungal diversity through morphological characters of spores and intraradicular hyphae has suggested previously that preferential associations occur between plants and AM fungi. A field experiment was established to investigate whether AM fungal diversity is affected by different host plants in upland grasslands. Indigenous vegetation from plots in an unimproved pasture was replaced with monocultures of either Agrostis capillaris or Lolium perenne. Modification of the diversity of AM fungi in these plots was evaluated by analysis of partial sequences in the large subunit (LSU) ribosomal RNA (rDNA) genes. General primers for AM fungi were designed for the PCR amplification of partial sequences using DNA extracted from root tissues of A. capillaris and L. perenne. PCR products were used to construct LSU rDNA libraries. Sequencing of randomly selected clones indicated that plant roots were colonised by AM fungi belonging to the genera Glomus, Acaulospora and Scutellospora. There was a difference in the diversity of AM fungi colonising roots of A. capillaris and L. perenne that was confirmed by PCR using primers specific for each sequence group. These molecular data suggest the existence of a selection pressure of plants on AM fungal communities.     

Rapid and reliable DNA extraction techniques from trypan-blue-stained mycorrhizal roots: comparison of two methods

Two improved DNA extraction techniques from trypan-blue-stained root fragments were developed and compared for rapid and reliable analyses. In Method A, 1 cm trypan-blue-stained mycorrhizal root fragments were individually isolated, crushed by bead beating, and purified with Chelex-100 (Bio-Rad). In Method B, DNA extraction was carried out using an UltraClean microbial DNA isolation kit (MoBio Laboratories). DNA was extracted from the mycorrhizal roots of four plant species, quantified by UV absorbance, and PCR-amplified with primers specific to arbuscular mycorrhizal fungi. Although PCR inhibitors might still exist when using Method A, appropriate dilution and employment of nested-PCR overcame this problem. Method B removed PCR inhibitors, but sometimes, depending on the mycorrhizal colonization within the root fragments, it also required nested PCR. In conclusion, both methods enabled us to handle many samples in a short time. Method B provided greater reliability and Method A provided better cost performance. Both techniques can be useful for PCR-based applications to identify species and estimate species composition after measuring mycorrhizal colonization rate with trypan blue staining.     

Arbuscular mycorrhizal fungal propagules in a salt marsh

The tolerance of indigenous arbuscular mycorrhizal fungi (AMF) to stressful soil conditions and the relative contribution of spores of these fungi to plant colonization were examined in a Portuguese salt marsh. Glomus geosporum is dominant in this salt marsh. Using tetrazolium as a vital stain, a high proportion of field-collected spores were found to be metabolically active at all sampling dates. Spore germination tests showed that salt marsh spores were not affected by increasing levels of salinity, in contrast to two non-marsh spore isolates, and had a significantly higher ability to germinate under increased levels of salinity (20) than in the absence of or at low salinity (10). Germination of salt marsh spores was not affected by soil water levels above field capacity, in contrast to one of the two non-marsh spore isolates. For the evaluation of infectivity, a bioassay was established with undisturbed soil cores (containing all types of AM fungal propagules) and soil cores containing only spores as AM fungal propagules. Different types of propagules were able to initiate and to expand the root colonization of a native plant species, but spores were slower than mycelium and/or root fragments in colonizing host roots. The AM fungal adaptation shown by this study may explain the maintenance of AMF in salt marshes.     

Arbuscular mycorrhizal fungi colonize decomposing leaves of<Emphasis Type="Italic"> Myrica parvifolia</Emphasis>,<Emphasis Type="Italic"> M. pubescens</Emphasis> and<Emphasis Type="Italic"> Paepalanthus</Emphasis> sp.

Hyphae and vesicles of arbuscular mycorrhizal fungi (AMF) were found within the decomposing leaves of Myrica parvifolia, M. pubescens and Paepalanthus sp. at three montane sites in Colombia. Hyphae, vesicles, and arbuscule-like structures were also found within scale-like leaves of the rhizomes of Paepalanthus sp. The litter found in the vicinity of the roots was divided into three decomposition layers. The highest AMF colonization occurred in the most decomposed leaves, which were in close association with roots. In contrast, there were no differences in AMF colonization of roots present in the different decomposition layers. Colonization of decomposing leaves by AMF did not differ between the two closely related species M. parvifolia and M. pubescens, nor between two sites (Guatavita and Zipacón, Colombia) differing in soil fertility. Occurrence of vesicles in decomposing leaves was correlated with abundant AMF extraradical hyphae among the leaves. We propose that AMF enter decomposing leaves mechanically through vascular tissue. As a consequence, AMF are well positioned to obtain and efficiently recycle mineral nutrients released by decomposer microorganisms before their loss by leaching or immobilization in soil.     

Element distribution in mycorrhizal and nonmycorrhizal roots of the halophyte Aster tripolium determined by proton induced X-ray emission

Summary. The salt aster (Aster tripolium L.) colonized by the arbuscular mycorrhizal fungus Glomus intraradices Sy167 and noncolonized control plants were grown in a greenhouse for nine months with regular fertilization by Hoagland nutrient solution supplemented with 2% NaCl. Mycorrhizal roots showed a high degree of mycorrhizal colonization of 60–70% and formed approximately 25% more dry weight and much less aerenchyma than the nonmycorrhizal controls. Cryosectioning essentially preserved the root cell structures and apparently did not cause significant ion movements within the roots during cuttings. The experimental conditions, however, did not allow to discriminate between fungal and plant structures within the roots. Quantification of proton-induced X-ray emission (PIXE) data revealed that in control roots, Na⁺ was mainly concentrated in the outer epidermal and exodermal cells, whereas the Cl^– concentration was about the same in all cells of the roots. Cross sections of roots colonized by the mycorrhizal fungus did not show this Na1 gradient in the concentration from outside to inside but contained a much higher percentage of NaCl among the elements determined than the controls. PIXE images are also presented for the four other elements K, P, S, and Ca. Both in colonized and control roots, the concentration of potassium was high, probably for maintaining homoeostasis under salt stress. This is seemingly the first attempt to localize both Na⁺ and Cl^– in a plant tissue by a biophysical method and also demonstrates the usefulness of PIXE analysis for such kind of investigation.     

Effect of arbuscular mycorrhizal fungi on phytoextraction by corn (Zea mays) of lead-contaminated soil

The role of arbuscular mycorrhizal fungi (AMF) in lead (Pb) uptake by corn (Zea mays) grown in soil supplemented with Pb was examined. Plants were subjected to four Pb levels: 0 (control); 10 (low); 100 (medium); and 500 mg L(-1) (high). At each Pb level, plants were grown in soil without and with fungicide (benomyl) (20 mg kg(-1)) to suppress AMF activity. Benomyl significantly reduced AMF colonization at high. medium, and zero Pb exposures. Benomyl application resulted in significantly lower concentrations of phosphorus in leaves at low and medium Pb exposures. The benomyl-treated plants had higher Pb and manganese concentrations in leaves than plants not treated with benomyl. In addition, benomyl-treated plants had generally lower concentrations of zinc and copper in leaves than plants not treated with benomyl. These results suggest that the role of AMF in heavy metal uptake is metal specific. Based on this work, the use of benomyl on soils contaminated with Pb can be recommended in phytoextraction.