Red list plants: colonization by arbuscular mycorrhizal fungi and dark septate endophytes

Since information concerning the mycorrhization of endangered plants is of major importance for their potential re-establishment, we determined the mycorrhizal status of Serratula tinctoria (Asteraceae), Betonica officinalis (Lamiaceae), Drosera intermedia (Droseraceae) and Lycopodiella inundata (Lycopodiaceae), occurring at one of two wetland sites (fen meadow and peat bog), which differed in soil pH and available P levels. Root colonization by arbuscular mycorrhizal fungi (AMF) and dark septate endophytes (DSE) was quantified. Colonization by AMF appeared to be more frequent in the fen meadow than in the peat bog, and depended on the host plant. Roots of S. tinctoria and B. officinalis were well colonized by AMF in the fen meadow (35–55% root length) and both arbuscules and vesicles were observed to occur in spring as well as in autumn. In the peat bog, L. inundata showed a low level of root colonization in spring, when vesicles were found frequently but no arbuscules. In roots of D. intermedia from the peat bog, arbuscules and vesicles were observed, but AMF colonization was lower than in L. inundata. In contrast, the amount of AMF spores extracted from soil at the peat bog site was higher than from the fen meadow soil. Spore numbers did not differ between spring and autumn in the fen meadow, but they were higher in spring than in autumn in the peat bog. Acaulospora laevis or A. colossica and Glomus etunicatum were identified amongst the AMF spores extracted from soil at the two sites. S. tinctoria and B. officinalis roots were also regularly colonized by DSE (18–40% root length), while L. inundata was only rarely colonized and D. intermedia did not seem to be colonized by DSE at all.     

Impact of dark septate endophytes on tomato growth and nutrient uptake

ABSTRACT

Background

Dark septate endophytes (DSEs) represent a form-group of ascomycetous fungi that inhabit the roots of a wide range of plant species, but our knowledge on their interaction with the host plants is still limited.     

Wetland dicots and monocots differ in colonization by arbuscular mycorrhizal fungi and dark septate endophytes

As an initial step towards evaluating whether mycorrhizas influence composition and diversity in calcareous fen plant communities, we surveyed root colonization by arbuscular mycorrhizal fungi (AMF) and dark septate endophytic fungi (DSE) in 67 plant species in three different fens in central New York State (USA). We found colonization by AMF and DSE in most plant species at all three sites, with the type and extent of colonization differing between monocots and dicots. On average, AMF colonization was higher in dicots (58±3%, mean±SE) than in monocots (13±4%) but DSE colonization followed the opposite trend (24±3% in monocots and 9±1% in dicots). In sedges and cattails, two monocot families that are often abundant in fens and other wetlands, AMF colonization was usually very low (<10%) in five species and completely absent in seven others. However, DSE colonization in these species was frequently observed. Responses of wetland plants to AMF and DSE are poorly understood, but in the fen communities surveyed, dicots appear to be in a better position to respond to AMF than many of these more abundant monocots (e.g., sedges and cattails). In contrast, these monocots may be more likely to respond to DSE. Future work directed towards understanding the response of these wetland plants to AMF and DSE should provide insight into the roles these fungal symbionts play in influencing diversity in fen plant communities.     

Seasonality of arbuscular mycorrhizal symbiosis and dark septate endophytes in a grassland site in southwest China

Arbuscular mycorrhizal and dark septate endophytic fungal colonization in a grassland in Kunming, southwest China, was investigated monthly over one year. All plant roots surveyed were co-colonized by arbuscular mycorrhizal and dark septate endophytic fungi in this grassland. Both arbuscular mycorrhizal and dark septate endophytic fungal colonization fluctuated significantly throughout the year, and their seasonal patterns were different in each plant species. The relationships between environmental (climatic and edaphic) factors and fungal colonization were also studied. Correlation analysis demonstrated that arbuscular mycorrhizal colonization was significantly correlative with environmental factors (rainfall, sunlight hours, soil P, etc.), but dark septate endophytic fungal colonization was only correlative with relative humidity and sunlight hours.     

荒漠北沙柳根系丛枝菌根真菌和黑隔内生真菌定殖状况

为利用土壤共生真菌资源促进荒漠植被恢复和生态重建, 分别于2013年6月、8月和10月, 从内蒙古元上都地区采集北沙柳(Salix psammophila)根围0-10、10-20、20-30、30-40和40-50 cm共5个土层的土壤样品, 系统研究了丛枝菌根真菌(AMF)和黑隔内生真菌(DSE)的时空分布及其与土壤因子的相关性。结果表明: AMF和DSE的平均定殖率分别为77%和84%, 说明北沙柳根系能与这两类真菌形成良好的共生关系。AMF和DSE的分布和定殖具有明显的时空异质性, 并与土壤因子密切相关。AMF和DSE的平均定殖率均表现为10月> 8月> 6月。土壤深度对AMF和DSE的定殖率有显著影响, AMF和DSE定殖率的最大值分别在0-20 cm和0-10 cm土层。双因子方差分析表明, 月份和土层对AMF和DSE的定殖率以及土壤因子具有显著的交互效应。主成分分析表明, 土壤湿度、pH值、碱性磷酸酶、易提取球囊霉素是内蒙古荒漠环境中AMF和DSE定殖的主要影响因子。     

丛枝菌根真菌与深色有隔内生真菌生态修复功能与作用

生态修复是目前全球关注的热点问题,如何增加植被的覆盖度及生态修复效率是目前研究的重点。丛枝菌根真菌(arbuscular mycorrhiza fungi,AMF)和深色有隔内生真菌(dark septate endophyte,DSE)均是自然界植物根际分布广泛的一类内生真菌,均能与植物形成菌根共生体,具有一定的促进植物生长、抵抗逆境及修复污染土壤等功能与作用,在生态修复中具有广泛的应用潜力。本文综述了AMF及DSE两种微生物的功能、作用及其在生态修复应用中的研究进展,并进一步对AMF和DSE在生态修复中存在的问题和前景进行展望。     

Occurrence and coexistence of arbuscular mycorrhizal fungi and dark septate fungi in aquatic macrophytes in a tropical river-floodplain system

In this study, carried out in four water bodies in the Upper Paraná River floodplain, we assessed the occurrence of root colonization by arbuscular mycorrhizal fungi (AMF) and dark septate fungi (DSF), as well as the AMF species richness associated with 24 species of aquatic macrophytes belonging to different life forms. AMF were found in nine species of macrophytes and DSF in 16 species among the 24 investigated. AM colonization occurred mainly in eudicotiledons (five of the six species evaluated) and the Paris morphology was the most common type. Co-occurrence of AMF and DSF was observed in seven species of macrophytes (Commelinaceae sp. 1, Limnobium laevigatum (H.B.K. ex Willd) Heine, Hygrophila cf. costata, Myriophyllum brasiliense (Camb), Polygonum acuminatum Kunth, P. ferrugineum Wedd and P. stelligerum Cham). Four species of macrophytes (Pistia stratiotes L., Eichhornia crassipes (Mart.) Solms, Egeria najas Planch and Nymphaea amazonum Mart. & Zucc) were not colonized by any type of fungi. In total, 27 morphotypes of AMF were recorded, and spores occurred both in the rhizosphere of macrophytes whose roots were internally colonized by AMF and in non-colonized macrophytes. Acaulospora delicata, Acaulospora aff. laevis, Acaulospora longula, Glomus lamellosum, Glomus luteum and NID 1 (a non-identified species) were the most frequent species. Samples collected close to the roots of N. amazonum had the highest AMF richness (20 species), but this plant was not colonized by fungi. A species richness curve indicated that more root-associated fungi than reported here are likely present in this floodplain.     

丛枝菌根真菌与深色有隔内生真菌对姜瘟病的防效及抗病机理初探

【背景】青枯劳尔氏菌(Ralstonia solanacearum,R.S)引发的姜瘟病是生姜产业发展的瓶颈问题。丛枝菌根真菌(arbuscular mycorrhiza fungi, AMF)与深色有隔内生真菌(dark septate endophytes,DSE)是两类重要的共生微生物。【目的】前期研究发现,AMF与DSE可提高生姜对姜瘟病的抗性,但其抗病机制尚不清楚,极大地限制了利用这两类共生真菌对该病的防治。【方法】在温室条件下做盆栽试验,以生姜组培苗为材料,设立接种AMF、DSE和不接种AMF、DSE的对照(CK)处理,并在上述处理下的植物生长4周后淋入病原菌液,病原菌接种1周后,通过测定菌根侵染率、发病率、叶绿素含量、光合指标、磷(P)含量、防御性酶活性及丙二醛(malondialdehyde, MDA)含量,研究AMF和DSE互作对病原菌侵染后生姜生长和生理生化指标的影响。【结果】AMF和DSE分别使姜瘟病发病率降低了45.27%和52.04%(P<0.05)。AMF+DSE组合处理抑病效果更好,发病率较对照降低60.87%(P<0.05)。AMF、DSE及...     

Spatial dynamics of dark septate endophytes in the roots and rhizospheres of Hedysarum scoparium in northwest China and the influence of edaphic variables

Spatial dynamics of DSE in the roots and rhizospheres of Hedysarum scoparium Fisch. et Mey. and soil factors were investigated at seven arid and semi-arid locations in northwest China in July 2015. Sampling sites were found to have a significant influence on the morphology, components, distribution, and infection of DSE in the roots of H. scoparium. Of nine DSE species isolated from H. scoparium roots, five are reported here for the first time from desert ecosystems. Hyphal infection in roots was positively correlated with soil urease and phosphatase activity. The presence of microsclerotia in roots was positively correlated with soil ammonium, and negatively correlated with soil organic matter and pH. We conclude that DSE infection is spatially predictable, and is influenced by nutrient availability and enzymatic activity. This research provides a basis for further understanding the ecological functions of DSE, and their roles in the promotion of vegetation restoration and in reducing erosion and desertification in arid ecosystems.     

宁夏枸杞深色有隔内生真菌的侵染特征及多样性

【背景】枸杞是享誉中外的名贵药材,深色有隔内生真菌(Dark septate endophytes,DSE)是枸杞内生真菌的重要组成部分。【目的】从宁夏枸杞栽培品种和野生品种的根系分离获得DSE菌株,研究枸杞DSE的群落组成、物种多样性及在宿主植物内的侵染定殖情况。【方法】从宁夏枸杞栽培园采集栽培品种宁杞1号、宁杞3号、宁杞5号、宁杞6号、宁杞7号、宁杞8号以及野生品种黄果枸杞和黑果枸杞共8个枸杞品种的根系,分离DSE菌株,运用形态学特征、r DNA-ITS序列分析进行菌株鉴定,采用回接试验方法确定DSE真菌。【结果】DSE在枸杞根系能形成大量"微菌核"典型结构。从8个枸杞品种根系中共分离获得DSE菌株279株,分属于18个属,具有丰富的物种多样性。镰刀菌属(Fusarium)为各品种的共有属和优势属,相对频率最高达85%。Monosporascus、蓝状菌属(Talaromyces)和俄氏孔菌属(Earliella)为枸杞内首次报道的DSE。枸杞不同品种中DSE群落物种多样性指数、均匀度指数和Simpson指数差异显著。【结论】DSE在枸杞栽培品种及野生品种中具有丰富的生物多样性,能够与枸杞根系形成良好的共生关系,增强了枸杞对生态环境的适应性。     

Seasonal and temporal dynamics of arbuscular mycorrhizal and dark septate endophytic fungi in a tallgrass prairie ecosystem are minimally affected by nitrogen enrichment

Root colonization by arbuscular mycorrhizae (AM) and dark septate endophytic (DSE) fungi in nitrogen amended and unamended mixed tallgrass prairie communities were analyzed monthly over two growing seasons. Roots were stained with Trypan blue and Sudan IV and fungal structures quantified using the modified magnified intersections method. Root length colonized (RLC) by DSE exceeded AM colonization during early part of the growing season. Fungal colonization varied among the years and was greater in 2003 than in 2002. Seasonal variation among the months within a growing season was observed in 2002 but not in 2003 for both AM and DSE. AM fungi were most abundant during the peak growing season of dominant C₄ vegetation while DSE were most abundant during the early part of the growing season. Hyperparasitism of AM hyphal coils by melanized septate fungi was frequently observed and increased with AM coil frequency. Although nitrogen amendment had altered the plant community composition, it had no impact on the colonization by AM or DSE fungi.     

西北荒漠带细枝岩黄耆AM真菌与DSE真菌时空分布研究

为探明细枝岩黄耆Hedysarum scoparium根系丛枝菌根真菌（arbuscular mycorrhizae,AM）和深色有隔内生真菌（dark septate endophytes,DSE）定殖特征及其生态地理分布,充分理解菌根共生体在植物生长和植被恢复中的功能,本研究连续3年采集我国西北荒漠带不同样地细枝岩黄耆0-30cm土层土壤样品和根样,不同样地细枝岩黄耆均能被AM和DSE侵染,形成AM典型结构菌丝、泡囊和丛枝,DSE典型结构深色有隔菌丝和微菌核。细枝岩黄耆根系AM真菌定殖高于DSE定殖,但极端干旱条件下DSE定殖优于AM真菌定殖。AM真菌定殖率不同年际间表现为2016年最高,同一年份不同样地,除乌海、沙坡头样地,菌丝定殖率自东向西呈降低趋势。DSE真菌定殖率,在同一样地不同年份：乌海样地2016年定殖率显著高于2015年和2017年,在同一年份不同样地：2015年沙坡头样地DSE菌丝定殖率、总定殖率显著高于其他样地,而2016、2017年鄂尔多斯样地最高。NMDS分析结果显示,AM和DSE真菌定殖不同年际间差异显著。相关性分析表明,AM真菌定殖率与DSE定殖率显著正相关;土壤温度与AM定殖显著负相关;有机碳、速效磷、磷酸酶、湿度与AM定殖显著正相关;DSE菌丝定殖率、定殖强度仅与酸性磷酸酶正相关。本研究比较分析西北荒漠带不同样地AM和DSE定殖与土壤因子的相关性,有助于充分理解菌根共生体在植物生长和植被恢复中的功能和意义。     

入侵植物五爪金龙根系深色有隔内生真菌的侵染特征

深色有隔内生真菌（DSE）广泛侵染华南地区外来入侵植物五爪金龙。显微观察发现五爪金龙根系具有大量的泡囊,其最显著的特征是大量脂质的富集和被分解成小脂质体,并通过透明或深色菌丝转移,说明了五爪金龙和DSE之间进行碳转移。富含脂质的泡囊可产生透明菌丝和自我复制,说明泡囊具有繁殖功能。富含脂质的泡囊和透明菌丝等非典型结构没有可分辨的或具透明的细胞壁,是活跃的真菌结构。透明菌丝和富含脂质的泡囊主要定殖在生理活性强的根组织中。深色有隔菌丝和微菌核是DSE的典型结构,主要定殖在相对不活跃的根组织中。DSE定殖于五爪金龙根的维管柱、皮层、表皮细胞和表面,同时延伸到土壤,在宿主植物和土壤之间形成连续的系统的真菌网络。根据DSE的多态性质、庞大的真菌网络和脂质的富集及其分布,探讨了DSE和入侵植物五爪金龙潜在的互惠共生关系。     

Species composition and colonization of dark septate endophytes are affected by host plant species and soil depth in the Mu Us sandland,northwest China

Investigations into the edaphic associations, host affiliations and soil depth of dark septate endophytes (DSE) in arid desert environments can help explain their spatial distribution and the response mechanisms in desert ecosystems. Soils were sampled to a depth of 50 cm in the rhizospheres of Psammochloa villosa, Hedysarum laeve and Artemisia ordosica in the Mu Us sandland of northwest China in July 2015. The plant species and soil depth significantly influenced the distribution and colonization of DSE. Hyphal and total root colonization were significantly higher under P. villosa than the others in the 0–20 cm layer. The maximum colonization of P. villosa and H. laeve occurred in the 10–20 cm and 20–30 cm soil layers, respectively, while 30–40 cm soil layer under A. ordosica. Of twelve DSE species isolated from the roots of these plants, Phoma radicina and Bipolaris zeae were reported in desert ecosystems for the first time. Hyphal colonization was significantly and positively correlated with soil total nitrogen (TN) and significantly and negatively correlated with the soil carbon/nitrogen (C/N) ratio. Microsclerotial colonization was significantly and positively correlated with soil organic carbon (SOC), and total colonization was significantly and positively correlated with soil TN and total phosphorus (TP) and significantly and negatively correlated with soil C/N. Variation of DSE colonization was mostly attributed to effects of plant species. We concluded that the species composition and colonization of the DSE fungi were influenced by the plant species, soil depth and soil nutrient availability in this desert ecosystem. This research provides a basis for further understanding the ecological adaptability of DSE and their roles in promoting vegetation restoration and reducing desertification in arid ecosystems.     

Are lulworthioid fungi dark septate endophytes of the dominant Mediterranean seagrass Posidonia oceanica?

• A previous study from Sicily, Italy, indicated that the dominant Mediterranean seagrass Posidonia oceanica forms a dark septate endophytic (DSE) association with a lulworthioid fungus (‘Lulwoana sp.’). This conflicts with several other studies from the NW Mediterranean Sea that point at the recently described pleosporalean fungus Posidoniomyces atricolor.
• I collected P. oceanica roots at eight sites around Sicily and checked them for fungal colonization using light microscopy. At three sites, root fungal symbionts (mycobionts) were isolated into pure cultures and identified using ITS rDNA sequences.
• Posidoniomyces atricolor represented the most frequent mycobiont (56 isolates), closely followed by lulworthioid fungi (51). The obtained mycobiont spectrum also included Cladosporium (2), Alternaria (1), Corollospora (1), Fusarium (1), Penicillium (1) and Vishniacozyma (1) isolates. The characteristic DSE root colonization, similar to those occurring in terrestrial plants but not known from any other seagrass, was found in all investigated P. oceanica individuals. The microscopy screening suggests that P. atricolor is responsible for the observed DSE colonization.
• This study extends the known range of Pos. atricolor and the DSE association characteristic for P. oceanica to the southern Tyrrhenian Sea/Sicily. While lulworthioid fungi regularly occur in P. oceanica tissues, including terminal fine roots, their significance and functioning (e.g. parasitic, pathogenic, endophytic) are unknown and require further investigation. However, there is currently no proof that they belong among dark septate endophytes of this seagrass.

     

深色有隔内生真菌不同培养时间对胞外聚合物合成的影响

【目的】探索不同培养时间下深色有隔内生真菌(CGMCC17463)合成胞外聚合物(extracellular polymeric substance, EPS)的产量及组成变化。【方法】通过摇瓶培养深色有隔内生真菌(dark septate endophytes, DSE),分析不同培养时间合成的EPS产量及应用的差异。【结果】摇瓶发酵试验表明,第4-12天,DSE生长趋于对数期和稳定期,EPS合成速率较快,在第12天产量达到1.41 g/L,之后EPS的合成速率逐渐降低。EPS组分分析结果表明,在相同质量的EPS中,第12天的胞外多糖含量较高,随着DSE生长进入稳定期至衰亡期,菌丝裂解,EPS中蛋白含量显著提高。官能团分析结果表明,不同培养时间的EPS中官能团种类并未发生变化,仅存在官能团含量上的改变。扫描电子显微镜和粒径分析结果表明,随着培养时间的增加,EPS的结构逐渐发生改变,EPS中分子粒径小于5μm所占比例逐渐提高,分子粒径大于100μm所占比例逐渐降低。另外,EPS的生物活性测定结果表明,不同培养时间合成的EPS均具有清除氧自由基和保水的能力,并且这种能力受EPS组分相对丰...     

Dark septate endophyte and arbuscular mycorrhizal status of vegetation colonizing a bottomland hardwood forest after a 100 year flood

Levels of arbuscular mycorrhizal (AM) colonization and dark septate endophyte (DSE) colonization were assessed in the vegetation recolonizing a remnant bottomland hardwood forest in north central Texas following a 100 year flood. Thirty seven plant species representing 21 dicotyledonous and 2 montocotyledonous families established following floodwater recession. AM and/or DSE were found in all species. AM colonization was found in 31 out of the 37 species assessed including both monocotyledonous families (Poaceae and Cyperaceae) and 17 out of 21 dicotyledonous families (Acanthaceae, Asteraceae, Boraginaceae, Cucurbitaceae, Euphorbiaceae, Lamiaceae, Loganiaceae, Lythraceae, Malvaceae, Onagraceae, Pedaliaceae, Ranunculaceae, Sapindaceae, Scrophulariaceae, Solanaceae, Verbanaceae and Violaceae). DSE were found in 31 out of 37 species assessed including both monocotyledonous families and 15 out of 21 dicotyledonous families (Amaranthaceae, Asteraceae, Boraginaceae, Brassicaceae, Cucurbitaceae, Euphorbiaceae, Lamiaceae, Lythraceae, Malvaceae, Pedaliaceae, Phytolaccaceae, Polygonaceae, Ranunculaceae, Sapindaceae, Scrophulariaceae and Violaceae). There were no detectable differences in AM or DSE colonization levels among wetland indicator groups (p > 0.05). Levels of DSE colonization were negatively correlated with vesicular colonization and hyphal colonization for the obligate wetland species. There were no other significant relationships between AM and DSE colonization detected. Spearman rank order correlation coefficients did not differ significantly among wetland indicatory category for any level of AM or DSE colonization.     

Symbiotic fungi in roots of Artemisia annua with special reference to endophytic colonizers

Abstract

Advances on plant–fungal interactions reveal that root symbiotic fungi actively modulate host growth, resistance response and secondary metabolism. Artemisia annua has been widely recognized as an important medicinal plant for artemisinin production, yet little is known about the fungal consortium associated with roots of A. annua. In this article, microscopic and culture-dependant methods were used to evaluate the identity and taxonomic affinities of root symbiotic fungi. Morphological evidence confirmed that arbuscular mycorrhizal fungi were dominant fungal group in naturally regenerated roots, but low colonization frequency in planted roots. Dark septate endophytes (DSEs) were easily found, which were characterized with dark pigmented hypha and a sclerotium-like structure in root cortex, and other endophytic fungi also occurred. A total of 36 isolates were recovered. Combined morphological and molecular identification (based on ITS sequences) determined 21 fungal taxa (genotype), which were placed into numerous lineages of Ascomycota. The best BLAST match indicated that almost half of total taxa were closely related to undescribed fungi, some of them may act as novel DSEs but experimental data were warranted. Interestingly, remarkable difference of fungal community associated with two types of roots was examined and no culturable fungi overlapped. Our findings provide some additional evidence that DSEs and other root endophytes may be as common as mycorrhizal fungi. Recovered fungi as raw materials for bioassay of endophytes-mediated promotion of artemisinin content in A. annua will be conducted in further research.     

The incidence of arbuscular mycorrhiza in two submerged Isoëtes species

We investigated the occurrence of arbuscular mycorrhizal fungi in the roots of Isoëtes lacustris and I. echinospora. These submerged lycopsids are the only macrophyte species inhabiting the bottom of two acidified glacial lakes in the Czech Republic. Arbuscular mycorrhizal (AM) fungi were detected in the roots of both species but the percentage of root colonization was both low and variable. Nevertheless, planting Littorella uniflora in the sediments from Isoëtes rhizosphere revealed high levels of viable AM propagules in both lakes. While AM colonization of Isoëtes roots did not exceed 25%, the average colonization of Littorella roots amounted to more than 80%. Although colonization of quillwort roots by AM fungi is evident, the taxonomic identity and role of these AM fungi in plant growth remain unclear. In addition to AM fungi, root-colonizing dark septate endophytic fungi were observed in both Isoëtes species.