Mycorrhiza Reduces Adverse Effects of Dark Septate Endophytes (DSE) on Growth of Conifers

Mycorrhizal roots are frequently colonized by fungi of the Phialocephala fortinii s.l. - Acephala applanata species complex (PAC). These ascomycetes are common and widespread colonizers of tree roots. Some PAC strains reduce growth increments of their hosts but are beneficial in protecting roots against pathogens. Nothing is known about the effects of PAC on mycorrhizal fungi and the PAC-mycorrhiza association on plant growth, even though these two fungal groups occur closely together in natural habitats. We expect reduced colonization rates and reduced negative effects of PAC on host plants if roots are co-colonized by an ectomycorrhizal fungus (ECM). Depending on the temperature regime interactions among the partners in this tripartite ECM-PAC-plant system might also change. To test our hypotheses, effects of four PAC genotypes (two pathogenic and two non-pathogenic on the Norway spruce), mycorrhization by Laccaria bicolor (strain S238N) and two temperature regimes (19°C and 25°C) on the biomass of the Douglas-fir (Pseudotsuga menziesii) and Norway spruce (Picea abies) seedlings were studied. Mycorrhization compensated the adverse effects of PAC on the growth of the Norway spruce at both temperatures. The growth of the Douglas-fir was not influenced either by PAC or mycorrhization at 19°C, but at 25°C mycorrhization had a similar protective effect as in the Norway spruce. The compensatory effects probably rely on the reduction of the PAC-colonization density by mycorrhizae. Temperature and the PAC strain only had a differential effect on the biomass of the Norway spruce but not on the Douglas-fir. Higher temperature reduced mycorrhization of both hosts. We conclude that ectomycorrhizae form physical and/or physiological barriers against PAC leading to reduced PAC-colonization of the roots. Additionally, our results indicate that global warming could cause a general decrease of mycorrhization making primary roots more accessible to other symbionts and pathogens.     

Colonization Characteristics and Composition of Dark Septate Endophytes (DSE) in a Lead and Zinc Slag Heap in Southwest China

The colonization characteristics and diversity of dark septate endophytes (DSE) associated with plants growing in a lead and zinc slag heap with a more than 250-year history in southwestern China were assessed. A total of 117 plant samples belonging to 29 species and 19 families were examined, and 97% (28/29) were colonized by dark septate fungal hyphae or microsclerotia. DSE colonization intensity varied greatly across plant species and also across individuals of the same species, showing a hyphal colonization range of 0.0–35.7%. In total, 38 melanized root-associated fungi with various morphological characteristics were isolated and 82% (31/38) of fungi were closely related to well-known DSE fungi, such as Exophiala spp., Phialophora (= Cadophora) spp., Phialocephala spp., and Leptodontidium spp. Among these, Phialophora and Exophiala were the dominant genera. Our results showed that DSE dominantly colonized the roots of the pioneer plants growing in such an ancient Pb-Zn slag heap. By contributing to metal tolerance and nutrient acquisition, DSE may be integral to the function of metal mine ecosystems.     

The Potential of Dark Septate Endophytes to Form Root Symbioses with Ectomycorrhizal and Ericoid Mycorrhizal Middle European Forest Plants

The unresolved ecophysiological significance of Dark Septate Endophytes (DSE) may be in part due to existence of morphologically indistinguishable cryptic species in the most common Phialocephala fortinii s. l.—Acephala applanata species complex (PAC). We inoculated three middle European forest plants (European blueberry, Norway spruce and silver birch) with 16 strains of eight PAC cryptic species and other DSE and ectomycorrhizal/ericoid mycorrhizal fungi and focused on intraradical structures possibly representing interfaces for plant-fungus nutrient transfer and on host growth response. The PAC species Acephala applanata simultaneously formed structures resembling ericoid mycorrhiza (ErM) and DSE microsclerotia in blueberry. A. macrosclerotiorum, a close relative to PAC, formed ectomycorrhizae with spruce but not with birch, and structures resembling ErM in blueberry. Phialocephala glacialis, another close relative to PAC, formed structures resembling ErM in blueberry. In blueberry, six PAC strains significantly decreased dry shoot biomass compared to ErM control. In birch, one A. macrosclerotiorum strain increased root biomass and the other shoot biomass in comparison with non-inoculated control. The dual mycorrhizal ability of A. macrosclerotiorum suggested that it may form mycorrhizal links between Ericaceae and Pinaceae. However, we were unable to detect this species in Ericaceae roots growing in a forest with presence of A. macrosclerotiorum ectomycorrhizae. Nevertheless, the diversity of Ericaceae mycobionts was high (380 OTUs) with individual sites often dominated by hitherto unreported helotialean and chaetothyrialean/verrucarialean species; in contrast, typical ErM fungi were either absent or low in abundance. Some DSE apparently have a potential to form mycorrhizae with typical middle European forest plants. However, except A. applanata, the tested representatives of all hitherto described PAC cryptic species formed typical DSE colonization without specific structures necessary for mycorrhizal nutrient transport. A. macrosclerotiorum forms ectomycorrhiza with conifers but not with broadleaves and probably does not form common mycorrhizal networks between conifers with Ericaceae.     

不同程度铅锌污染区丛枝菌根真菌和深色有隔内生真菌侵染特征

对陕西凤县铅硐山铅锌矿区4个不同程度铅锌污染样地植物根系的丛枝菌根真菌(AMF)和深色有隔内生真菌(DSE)进行了资源和分布调查。结果表明:(1)无铅锌污染的矿山上调查的15种植物中除黄连木没有检测到AMF侵染、栓皮栎和酸模不能被DSE侵染外,其他植物均能与AMF和DSE共生,但平均侵染率较低,仅分别为32.3%和25.9%;铅锌轻度污染的尾矿荒地和铅中度污染、锌重度污染的尾矿坝植物根系内AMF和DSE侵染率明显提高,AMF的平均侵染率分别为53.3%和68.3%,DSE的平均侵染率分别为38.6%和54.2%;铅锌重度污染的废弃冶炼厂样地植物AMF侵染率明显下降,平均只有17.6%,而DSE的侵染没有受重金属污染的抑制,仍达到60.3%。(2)4个样地植物根际土壤中AMF孢子分布不均匀,每克土的孢子密度在0.08～6.84个,平均为1.5个。对4个样地均有分布的狼牙刺、博落回、山蒿和秦岭风毛菊的AMF、DSE侵染状况调查发现,轻度和中度的铅锌污染能促进AMF与宿主共生关系的建立,而重度污染则显著抑制AMF侵染;AMF孢子密度没有呈现一定的规律性,且与AMF侵染率之间没有显著相关性;DSE侵染率与土壤锌含量存在显著正相关关系(r=0.505,P<0.05),而与土壤铅含量没有显著相关性。可见,AMF和DSE能够广泛存在于铅硐山铅锌矿区,尤其是DSE表现出较强的耐受性和适应性。     

Occurrence of Arbuscular Mycorrhizas and Dark Septate Endophytes in Hydrophytes from Lakes and Streams in Southwest China

In this study, the colonization of arbuscular mycorrhizas (AM) and dark septate endophytes (DSE) in 140 specimens of 32 hydrophytes collected from four lakes and four streams in southwest China were investigated. The arbuscular mycorrhizal fungi (AMF) and DSE colonization in these hydrophytes were rare. Typical AM structures were observed in one of the 25 hydrophytic species collected in lakes and six of the 17 species collected in streams. Spores of 10 identified AMF species and an unidentified Acaulospora sp. were isolated from the sediments. The identified AMF came from the four genera, Acaulospora, Gigaspora, Glomus and Scutellospora . Glomus and G. mosseae were the dominant genus and species respectively in these aquatic environments. The presence of DSE in hydrophytes was recorded for the first time. DSE occurred in one of the 25 hydrophyte species collected in lakes and three of the 17 species collected in streams. (© 2006 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

The Co-occurrence and Morphological Continuum Between Ericoid Mycorrhiza and Dark Septate Endophytes in Roots of Six European Rhododendron Species

Ericaceae associate with a wide spectrum of root mycobionts, but the most common are ascomycetous ericoid mycorrhizal fungi and dark septate endophytes (DSE), followed by basidiomycetous fungi and glomeracean arbuscular mycorrhizal fungi. We investigated distribution and morphological diversity of ericoid mycorrhizae (ErM), DSE associations, ectomycorrhizae (EcM) and arbuscular mycorrhizae (AM) in hair roots of six European native Rhododendron species and found that i) while EcM and AM were absent, ErM and DSE associations were simultaneously present in all screened plants; ii) their levels were negatively correlated, suggesting Ericaceae preference for certain root-fungus association in certain habitats; iii) the highest ErM colonization occurred at sites in southern and central Europe, while the highest DSE colonization was found in a subarctic site in northern Finland and in a subalpine site in the Carpathians, suggesting a latitudinal/altitudinal shift in Ericaceae root-fungus associations; iv) some mycelia could simultaneously form structures corresponding to ErM and DSE association, which occasionally resulted in a unique ectendomycorrhizal colonization comprising an intercellular parenchymatous net and intracellular hyphal coils. These results indicate frequent interactions between ErM fungi and DSE in roots of European rhododendrons and a morphological continuum between ErM and DSE associations. The new ectendomycorrhizal type deserves further investigation.     

植物响应金属胁迫重要蛋白质的细胞定位

土壤中的Hg2+、Pb2+、Cd2+、Cu2+、Zn2+、Mn2+、Fe2+/Fe3+等金属离子对植物体具有毒害作用。植物体通过定位于细胞壁、细胞膜,以及各种细胞器中的蛋白质应答重金属胁迫过程。植物体利用细胞壁蛋白质与重金属离子络合,细胞膜系统的转运蛋白与通道调节金属离子运输、细胞器或细胞质基质中的分子伴侣帮助蛋白质折叠等机制完成抑制金属离子进入体内或在体内的解毒等过程。综述了近年来植物响应金属离子胁迫重要蛋白质的细胞定位与作用机制。     

Susceptibility of Halobacteria to Heavy Metals

Sixty-eight halobacteria, including both culture collection strains and fresh isolates from widely differing geographical areas, were tested for susceptibility to arsenate, cadmium, chromium, cobalt, copper, lead, mercury, nickel, silver, and zinc ions by an agar dilution technique. The culture collection strains showed different susceptibilities, clustering into five groups. Halobacterium mediterranei and Halobacterium volcanii were the most metal tolerant, whereas Haloarcula californiae and Haloarcula sinaiiensis had the highest susceptibilities of the culture collection strains. Different patterns of metal susceptibility were found for all the halobacteria tested, and there was a uniform susceptibility to mercury and silver. All strains tested were multiply metal tolerant.     

念珠藻葛仙米生理生化特性对不同低温胁迫的响应

研究了不同培养温度下对念珠藻葛仙米的生长和相关生理生化特性的变化.结果表明,低温抑制葛仙米的生长,在2℃～5℃低温下其生长甚至完全停止;在10℃低温胁迫下,其生长延滞期变长,其后仍能保持正常生长.低温胁迫下,葛仙米光合系统II(PSII)的光合效率(Fv/Fm)的变化趋势与生长曲线相似;细胞电解质渗透率在低温胁迫初期尚能保持比较低的水平,但随着时间延长而升高;可溶性糖和还原性糖含量的变化则随胁迫加剧有不同程度升高.可溶性蛋白含量则在低温下保持比较低的水平.这些结果表明,低温对葛仙米的生长和光合作用具有抑制作用,但葛仙米也表现出了一定的抵抗低温胁迫的能力,并通过调节细胞膜的渗透性、重新合成低温适应调节物质(如总糖和可溶性糖等)来减少低温伤害,从而适应低温胁迫.     

Evidence for Biotrophic Lifestyle and Biocontrol Potential of Dark Septate Endophyte Harpophora oryzae to Rice Blast Disease

The mutualism pattern of the dark septate endophyte (DSE) Harpophora oryzae in rice roots and its biocontrol potential in rice blast disease caused by Magnaporthe oryzae were investigated. Fluorescent protein-expressing H. oryzae was used to monitor the colonization pattern. Hyphae invaded from the epidermis to the inner cortex, but not into the root stele. Fungal colonization increased with root tissue maturation, showing no colonization in the meristematic zone, slight colonization in the elongation zone, and heavy colonization in the differentiation zone. H. oryzae adopted a biotrophic lifestyle in roots accompanied by programmed cell death. Real-time PCR facilitated the accurate quantification of fungal growth and the respective plant response. The biocontrol potential of H. oryzae was visualized by inoculation with eGFP-tagged M. oryzae in rice. H. oryzae protected rice from M. oryzae root invasion by the accumulation of H₂O₂ and elevated antioxidative capacity. H. oryzae also induced systemic resistance against rice blast. This systemic resistance was mediated by the OsWRKY45-dependent salicylic acid (SA) signaling pathway, as indicated by the strongly upregulated expression of OsWRKY45. The colonization pattern of H. oryzae was consistent with the typical characteristics of DSEs. H. oryzae enhanced local resistance by reactive oxygen species (ROS) and high antioxidative level and induced OsWRKY45-dependent SA-mediated systemic resistance against rice blast.     

Differential Sensitivity of Lichens to Heavy Metals

Zinc, Cd and Cu inhibited photosynthesis in lichens containingcyanobacterial phycobionts at substantially lower concentrationsthan those causing decreased photosynthesis in lichens containingchlorophycean phycobionts. This distinction was not relatedto differences in total thallus concentrations of Zn, Mg, Caor K or to the quantity of Zn taken up to intracellular sites.When incubated with concentrated Zn solutions the chlorophyceanlichen Cladonia rangiformis accumulated more Zn on extracellularexchangeable sites than did the cyanobacterial lichen Peltigerahorizontalis. algae, cyanobacteria, chlorophyceae, lichens, heavy metal, photosynthetic inhibition, zinc, cadmium, copper, cation uptake     

Septate junctions in the cephalic epidermis of turbellarians (Bipalium)

Summary In the epidermis of turbellarians septate junctions of the pleated sheet type have been demonstrated in conventional thin sections and freeze fractured preparations. The structure of these junctions entirely agrees with that found in molluscs and arthropods.Financially supported by DFG (Sto 75/3,4; We 380/5)     

Heavy Metals in Mussels (Mytilus galloprovincialis) from Marmara Sea, Turkey

Marmara Sea is one of the main catching areas of mussels (Mytilus galloprovincialis) in Turkey, and a significant portion of the harvest has been exported mainly to European countries. In this study, Zn, Cu, Cd, Hg, and Pb in mussels from ten catching areas in Marmara Sea were analyzed to investigate health risks associated with consuming mussels. Mercury was not detected (<0.15 ppb) in any of the samples. The highest concentrations of Cu and Cd were 3.473 and 0.740 mg kg^?1 (wet weight, WW), respectively, well below the maximum permissible levels. All samples contained Zn higher than 50 mg kg^?1, while Pb was above the limits in the samples from stations 1, 4, 6, and 8. Mussels from Marmara Sea are safe regarding Cu, Cd, and Hg but may contain Zn and Pb above the permissible limits. However, metal contents of mussels from Marmara Sea are mostly lower than those of the regions in other areas of the world. It was concluded that Marmara Sea has a potential of being a safe source of mussels if industrial inputs somewhat reduced and controlled. Concentrations of heavy metals in mussels must be monitored comprehensively and periodically with respect to the consumer health.     

Morphological Response of Witchweed (Striga asiatica) to In Vitro Culture

Excised sorghum root segments (5–10 mm in length) werecultured for 50 d in four different liquid media containingmineral salts, vitamins, amino acids, glucose, and IAA. Theroots were removed and the remaining medium was solidified withan equal volume of warm 1–6% water agar. Dry unconditionedor conditioned Striga asiatica seeds were transferred to themedium. Some of the seeds germinated and developed into parasitic-typeseedlings. These seedlings had haustoria, tubercles, dense roothairs, branched shoots, and multiple shoot-borne adventitiousroots. The plumule pole developed into a shoot, but the radiclepole displayed only rudimentary development. On the same media,but which had not previously been used to grow sorghum roots,the seedlings displayed a well-developed radicle-derived rootsystem, but the plumule did not grow. Shoots began to appearon the roots only after 35–50 d of culture. These seedlingshad no haustoria, no tubercles, few or scattered root hairs,no shoot-borne adventitious roots and few shoot branches, andappeared to be non-parasitic-type seedlings. Shoots grew ina medium supplemented with IAA and kinetin, but did not in amedium containing NAA plus IBA. On replacement of glucose andIAA with sucrose and 2,4-D, respectively, Striga seeds germinated,and the heart-shaped embryos dedifferentiated into calli. Thecalli have been maintained by subculturing for over 9 months.The results demonstrated that a host signal, in addition tothose for germination and haustorium formation, is requiredfor further development. Moreover, morphogenesis of culturedS. asiatica is influenced by exogenous growth regulators. Key words: Striga asiatica, parasitic weeds, haustoria, Sorghum bicolor, in vitro culture     

Production of Catalases by Aspergillus niger Isolates as a Response to Pollutant Stress by Heavy Metals

Isolates of Aspergillus niger, selected from the coal dust of a mine containing arsenic (As; 400 mg/kg) and from the river sediment of mine surroundings (As, 1651 mg/kg, Sb, 362 mg/kg), growing in minimal nitrate medium in the phase of hyphal development and spore formation, exhibited much higher levels of total catalase activity than the same species from the culture collection or a culture adapted to soil contaminated with As (5 mg/L). Electrophoretic resolution of catalases in cell-free extracts revealed three isozymes of catalases and production of individual isozymes was not significantly affected by stress environments. Exogenously added stressors (As⁵⁺, Cd²⁺, Cu²⁺) at final concentrations of 25 and 50 mg/L and H₂O₂ (20 or 40 mM) mostly stimulated production of catalases only in isolates from mines surroundings, and H₂O₂ and Hg²⁺ caused the disappearance of the smallest catalase I. Isolates exhibited a higher tolerance of the toxic effects of heavy metals and H₂O₂, as monitored by growth, than did the strain from the culture collection.