The genus <Emphasis Type="Italic">Diaporthe</Emphasis>: a rich source of diverse and bioactive metabolites

The genus Diaporthe (asexual state: Phomopsis) comprises pathogenic, endophytic and saprobic species with both temperate and tropical distributions. Although species of Diaporthe have in the past chiefly been distinguished based on host association, studies have confirmed several taxa to have wide host ranges, suggesting that they move freely between hosts, frequently co-colonizing diseased or dead tissue, while some species are known to be host-specific. They are also very frequently isolated as endophytes of seed plants. Due to their importance as plant pathogens, the genus has been thoroughly investigated for secondary metabolites, including during screening programs aimed at the discovery of novel bioactive natural products, but the respective information has never been compiled. Therefore, we have examined the relevant literature to explore and highlight the major classes of metabolites of Diaporthe and their Phomopsis conidial states. These fungi predominantly produce a large number of polyketides, but cytochalasins and other types of commonly encountered fungal secondary metabolites are also predominant in some species. Interestingly, not a single metabolite which is also known from the host plant has ever been isolated as a major component from an endophytic Diaporthe strain, despite the fact that many of the recent studies were targeting endophytic fungi of medicinal plants.     

红树林内生真菌研究进展

红树林作为一种特殊的植物群落具有丰富的内生真菌资源,目前已分离鉴定的红树林真菌超过200种,成为海洋真菌的第二大类群,已报道的红树林内生真菌主要类群是链格孢霉(Alternaria)、曲霉(Aspergillus)、芽枝霉(Cladosporium)、炭疽菌(Colletotrichum)、镰孢霉(Fusarium)、拟青霉(Paecilomyces)、拟盘多毛孢(Pestalotiopsis)、青霉(Penicillium)、茎点霉(Phoma)、拟茎点霉(Phomopsis)、叶点霉(Phyllosticta)和木霉(Trichoderma)等.大部分红树林内生真菌具有较宽的宿主范围,极少数只有单一的宿主,不同红树林植物的内生真菌区系及优势种群有很大差异.红树林内生真菌的定殖因宿主植物不同部位、植株的年龄及季节和环境的变化明显不同.红树林内生真菌能产生多种代谢产物,具有抗菌、抗肿瘤等药用价值.红树林植物内生真菌的研究和开发具有重要意义.本文综述了红树林内生真菌的生物多样性及其分布、生物学功能和次生代谢产物等方面的研究进展.     

龙南钾矿区常见蕨类植物可培养内生真菌的多样性

植物内生真菌作为一类特殊的微生物资源,与宿主在长期的生态系统演化过程中形成了互惠共生关系,通过多样化途径来增强植物体的营养生理和抗性机能,对宿主植物产生多种有益生物学作用,在植物演替过程中具有重要的生态学意义。这种特殊微生物资源近年来倍受关注,而利用植物-真菌共生体强化植物在矿区逆境中生长,提高矿区生物修复效率是一个新的研究热点。为探明钾矿区不同蕨类植物内生真菌的物种多样性、群落组成以及生态分布规律,该研究以芒萁、狗脊、禾秆蹄盖蕨、海金沙、华中铁角蕨、井栏边草和乌蕨等植物为材料,采用组织分离、形态学鉴定等方法,对其内生真菌多样性进行分析。结果表明:从7种植物中共分离获得377株内生真菌,总分离率在2.50%~4.52%之间。经鉴定377个菌株隶属于链格孢属、曲霉属、枝孢菌属和轮枝孢属等25个分类单元,其中链格孢属、曲霉属、枝孢菌属和轮枝孢属等在所有被调查蕨类植物中都有分布,为优势属,共计为185株,占总株数的49.07%,但它们在每种植物的分布存在明显差异;7种蕨类植物内生真菌总定植率为叶高于根状茎(P0.05),多样性指数在0.502~0.867之间,但每种植物及其不同组织部位的内生真菌定植率和多样性指数存在一定的差异;从相似性分析来看,同一个钾矿区不同蕨类植物内生真菌菌群之间的相似性程度较低,相似性系数在0.189~0.587之间。该研究结果不仅丰富了植物内生真菌种质资源,而且为进一步开展植物内生真菌强化宿主植物在钾矿区生长适应机制的研究奠定了基础。     

云南元江干热河谷五种优势植物的内生真菌多样性

从蔓草虫豆（Atylosia scarabaeoides）、余甘子（Phyllanthus emblica）和黄花稔（Sida acuta）等5种云南元江干热河谷植物的525个组织块中,共分离得到内生真菌371株,内生真菌的分离频率在0.61～0.92之间,且所有植物叶内生真菌的分离频率都明显高于茎（P<0.05）。经形态学鉴定,内生真菌分属于拟茎点霉属（Phomopsis sp.）、离蠕孢属（Bipolaris sp.）和交链孢属（Alternaria sp.）等32个分类单元。拟茎点霉属为干热河谷植物优势内生真菌属,从所有被调查植物的茎叶中都分离得到该属真菌,且相对分离频率高达12.90%～50.54%。内生真菌群落组成的多样性和相似性分析结果表明,云南元江干热河谷植物内生真菌多样性偏低、宿主专一性较小。     

植物内生拟盘多毛孢的生物多样性

内生拟盘多毛孢是植物 (尤其是木本植物 )内生真菌的重要类群。分离自红豆杉的内生小孢拟盘多毛孢(Pestalotiopsismicrospora)能产生抗癌的代谢产物紫杉醇 ,因而引起了人们对内生拟盘多毛孢研究的兴趣。拟盘多毛孢内生于植物的根、茎、叶及繁殖体中 ,已鉴定的内生拟盘多毛孢有 2 3种。在已调查的植物中 ,内生拟盘多毛孢的定殖率有很大差异 ,在红树 (Rhizophoraapiculata)、椰子 (Cocosnucifer)、西藏红豆杉 (Taxuswallichiana)、茶梅 (Camelliasasanqua)、Fragraeabodenii和Cordemoyaintegrifolia等木本植物中 ,拟盘多毛孢是内生真菌的优势类群。一种拟盘多毛孢可内生于多种植物体内 ,一种植物也可有多种拟盘多毛孢内生 ,其分布与植物所处的生态环境和地域有关。在一定的宿主植物生理条件或外界环境条件下 ,某些内生拟盘多毛孢可转变为寄生或腐生。对植物内生拟盘多毛孢代谢产物的研究表明 ,拟盘多毛孢可产生多种次生代谢产物 ,包括抗癌物质 (紫杉醇和粗榧酸 )和抗菌物质 (环己烷类物质、粗榧酸和 pestaloside)。植物内生拟盘多毛孢的资源调查和开发应用具有广阔的前景。     

Diversity and ecological distribution of endophytic fungi associated with medicinal plants

A total of 973 isolates of endophytic fungi were recovered from 1144 tissue fragments of the six medicinal plant species belonging to 4 families collected in the Beijing Botanical Garden. Of these isolates 778 sporulated and were identified into 21 taxa by morphological characteristics. Among the taxa 11 belonged to Coelomycetes, 6 to Ascomycetes, and 4 to Hyphomycetes. Various numbers of endophytic fungi (5–8 taxa) were obtained from each plant. Alternaria alternata was the dominant species in the 6 plants, and Microsphaeropsis conielloides was also dominant in Eucommia ulmoides. There were high colonization rates (47.9%–63.1%) and isolation rates (0.7–0.93) of endophytic fungi, and they were conspicuously higher in twigs than those in leaves in the 6 plants examined. The colonization and isolation rates of endophytic fungi increased with the twig age. The results based on the analyses of cluster and Sorenson’s similarity coefficients indicated that some endophytic fungi showed a certain degree of host and tissue preference.     

Antiproliferative,Antifungal, and Antibacterial Activities of Endophytic Alternaria Species from Cupressaceae

Recent research has shown the bioprospecting of endophytic fungi from Cupressaceae. Here, we further uncover that the healthy cypress plants such as Cupressus arizonica, Cupressus sempervirens var. cereiformis, and Thuja orientalis host highly bioactive endophytic Alternaria fungal species. Indeed, endophytic Alternaria alternata, Alternaria pellucida, and Alternaria tangelonis were recovered from healthy Cupressaceous trees. Biodiversity and bioactivity of recovered endophytic Alternaria species were a matter of biogeography and host identity. We further extracted such Alternaria’s metabolites and highlighted their significant antiproliferative, growth inhibitory, and antibacterial activities against the model target fungus Pyricularia oryzae and the model pathogenic bacteria Bacillus sp., Erwinia amylovora, and Pseudomonas syringae. In vitro assays also indicated that endophytic Alternaria species significantly inhibited the growth of cypress fungal phytopathogens Diplodia seriata, Phaeobotryon cupressi, and Spencermartinsia viticola. In conclusion, since the recovered Alternaria species were originally reported as pathogenic and allergenic fungi, our findings suggest a possible ecological niche for them inside the foliar tissues of Cupressaceous trees. Moreover, in this study, the significant bioactivities of endophytic Alternaria species in association with Cupressaceae plant family are reported.     

Anatomical relations among endophytic holoparasitic angiosperms, autotrophic host plants and mycorrhizal fungi: A novel tripartite interaction

Mycorrhizae are widespread mutualistic symbioses crucial for the functioning of terrestrial ecosystems. Not all plants associate with mycorrhizae; most parasitic plants have been suggested to be nonmycorrhizal because they have developed alternative strategies to obtain nutrients. In endophytic parasitic plants, whose vegetative bodies grow completely inside their mycorrhizal host roots, the opportunity for establishing a tripartite association seems evident, but information on these systems is lacking. In studying natural associations among the endophytic holoparasite Cytinus hypocistis, their Cistaceae host species, and associated mycorrhizal fungi, we found that mycorrhizae were associated with the hosts and the parasites, reaching high frequencies of colonization. In parasitic and host root tissues, mycorrhizal fungi spread in the parenchymatic cells by intracellular growth and formed hyphal coils and vesicles, while the cambium and the vascular tissues were never colonized. This report is the first on a tripartite association of an endophytic parasitic plant, its host, and mycorrhizae in natural conditions, representing a novel trophic interaction not previously reported within the angiosperms. Additional studies on the interactions occurring among these three players are needed because they may be crucial to our understanding of how this mutualistic-antagonistic system is functioning and evolving.     

Fungal diversity living in the root and sporophore of the endemic Korean fern Mankyua chejuense

Ferns represent the basal group of vascular plants and are known to have fungal interactions with arbuscular mycorrhizal fungi, but diversity of endophytic fungi from ferns is rarely studied. Moreover, fungal diversity associated with ferns is likely underestimated as most studies have been performed based on a microscopic or culture-dependent approach. In this study, we investigated the endophytic fungal diversity within roots and sporophore of an endangered Korean fern (Mankyua chejuense), and compared it to fungi in surrounding soil using a metabarcoding approach. A high diversity of endophytic fungi (236 OTUs), mostly belonging to Ascomycota, was detected and fungal richness and composition were significantly different between habitats. Indicator species analysis showed that endophytic fungi have similar ecological characteristics to fungal species found from other land plants. Our results suggest that various fungal species are associated with ferns, thus understanding fern-associated fungal diversity can have a great implication for fern biology and conservation.     

云南会泽铅锌矿废弃矿渣堆常见植物内生真菌多样性

从云南会泽铅锌矿废弃矿渣堆上的常见植物硬毛南芥(Arabis hirsuta)、毛萼香茶菜(Rabbosia eriocalyx)和倒挂刺(Rosa longicuspis)等6种植物的690个组织块中共分离得到内生真菌495株,内生真菌的分离频率在0.42—0.93之间,平均为0.72,所有植物茎内生真菌的分离频率都明显高于叶(P<0.05)。经形态学鉴定,内生真菌分属于茎点霉属(Phoma)、交链孢属(Alternaria)和派伦霉属(Peyronellaea)等20个分类单元,其中茎点霉属和派伦霉属为该废弃矿渣堆上常见植物的优势内生真菌属。6种植物内生真菌的多样性指数在1.05—2.29之间,与其它非重金属污染环境植物内生真菌的多样性指数相似,说明在重金属污染地区仍然存在多种重金属耐受的内生真菌种类。6种植物内生真菌的相似性系数(0.455—0.833)表明,会泽铅锌矿区植物内生真菌的宿主专一性较小。     

Piperine production by endophytic fungus Colletotrichum gloeosporioides isolated from Piper nigrum

Many endophytic fungi have been reported with the biosynthetic potential to produce same or similar metabolites present in host plants. The adaptations that might have acquired by these fungi as a result of the long-term association with their host plants can be the possible basis of their biosynthetic potential. The bioactive compounds originated from endophytes are currently explored for their potential applications in pharmaceutical, agriculture and food industries. Piper nigrum, a plant of the Piperaceae is very remarkable because of the presence of the alkaloid piperine. Piperine has been reported to have broad bioactive properties ranging from antimicrobial, antidepressant, anti-inflammatory, antioxidative to anticancer activities. Interestingly, piperine also plays a vital role in increasing the bioavailability of many drugs which again is a promising property. The current study was carried out to identify piperine producing endophytic fungus from Piper nigrum L. By screening various endophytic fungi, the isolate which was identified as member of Colletotrichum gloeosporioides was found to have the ability to form piperine and was confirmed by HPLC and LCMS. Considering the broad bioactive potential of piperine, the piperine producing fungi identified in the study can expect to have much industrial potential.     

Fungal endophytes from cactus Cereus jamacaru in Brazilian tropical dry forest: a first study

Endophytic fungi live within the healthy tissues of plants and can promote host species tolerance to different environmental stresses. However, most studies have been of plants in humid environments, and there are few reports of the benefits of such associations for plants of extreme environments. Our aims were to identify endophytic fungi using morphological taxonomy, to explore richness and estimate species frequency in a cactus, C. jamacaru, in Brazilian tropical dry forest (Caatinga). We thus identified 59 taxa, corresponding to 69.7 % of the total number of isolates; the other 30.3 % were sterile mycelia. Cladosporium cladosporioides and Fusarium oxysporum were the species most commonly isolated, followed by Acremonium implicatum, Aureobasidium pullulans, Trichoderma viride, Chrysonilia sitophila, and Aspergillus flavus. Forty-seven species are recorded for the first time as endophytic fungi of cacti, and 18 others as endophytes for Brazil; this suggests that C. jamacaru harbors a highly diverse fungal community as measured by a diversity index. However, species accumulation curves suggest that our study still underestimates endophyte diversity because it does not provide an exhaustive sample. To our knowledge, this is the first report of endophytic fungi from C. jamacaru in tropical dry forests.     

Diversity and ecological distribution of endophytic fungi associated with medicinal plants

A total of 973 isolates of endophytic fungi were recovered from 1144 tissue fragments of the six me-dicinal plant species belonging to 4 families collected in the Beijing Botanical Garden. Of these isolates 778 sporulated and were identified into 21 taxa by morphological characteristics. Among the taxa 11 belonged to Coelomycetes, 6 to Ascomycetes, and 4 to Hyphomycetes. Various numbers of endophytic fungi (5―8 taxa) were obtained from each plant. Alternaria alternata was the dominant species in the 6 plants, and Microsphaeropsis conielloides was also dominant in Eucommia ulmoides. There were high colonization rates (47.9%―63.1%) and isolation rates (0.7―0.93) of endophytic fungi, and they were conspicuously higher in twigs than those in leaves in the 6 plants examined. The colonization and isolation rates of endophytic fungi increased with the twig age. The results based on the analyses of cluster and Sorenson's similarity coefficients indicated that some endophytic fungi showed a certain degree of host and tissue preference.     

Community of Endophytic Fungi from Alpine Conifers on Mt. Seorak

Endophytic fungi occupy various ecological niches, which reinforces their diversity. As few studies have investigated the endophytic fungi of alpine conifers, we focused on four species of alpine conifers in this study—Abies nephrolepis, Pinus pumila, Taxus cuspidata var. nana, and Thuja koraiensis—and examined them for endophytic fungi. A total of 108 endophytic fungi were isolated. There were four taxa in A. nephrolepis, 12 in P. pumila, 18 in T. cuspidata var. nana, and 17 in T. koraiensis; these were divided into five classes: Agaricomycetes (3.2%), Dothideomycetes (29.0%), Leotiomycetes (15.0%), Sordariomycetes (41.9%), and Orbiliomycetes (1.6%). The most prevalent fungi were Sydowia polyspora (22.7%) and Xylariaceae sp. (22.7%) in P. pumila, Phomopsis juglandina (16.1%) in T. cuspidata var. nana, and Thuja-endophytes sp. 1 (70.0%) in T. koraiensis. However, there was no dominant species growing in A. nephrolepis. Some host plants were analyzed using next-generation sequencing. We obtained 4618 reads for A. nephrolepis and 2268 reads for T. koraiensis. At the genus level, the top three endophytic fungi were Ophiostomataceae_uc (64.6%), Nectriaceae_uc (15.5%), and unclassified organism (18.0%) in A. nephrolepis and Nectriaceae_uc (41.9%), Ophiostomataceae_uc (41.8%), and Magnaporthaceae_uc (9.2%) in T. koraiensis. Our results show that there are different communities of endophytic fungi among different host plants, even if the host plants are in the same region. Such ecological niches are important in terms of the ecological restoration of alpine conifers.     

Fungal endophytes of wild barley and their effects on Diuraphis noxia population development

Laboratory experiments were conducted to compare the expression of Diuraphis noxia (Mordvilko) (Homoptera: Aphididae) resistance in four plant introduction (PI) lines of wild barley (Hordeum) infected with different species or strains of endophytic fungi (tribe Balansieae, family Clavicipitaceae, Neotyphodium gen. nov. [formerly Acremonium]). Aphid densities were significantly lower on endophyte-infected plants of PI 314696 (H. bogdanii Wilensky) and PI 440420 (H. brevisubulatum subsp. violaceum (Boissier & Hohenacker)), compared with densities on endophyte-free plants of both PI lines in population growth experiments. This endophyte- associated resistance was the result of antibiosis effects or starvation. In other experiments, endophyte-free plants of PI 269406 and PI 440413 (H. bogdanii) were not superior to endophyte-infected conspecifics as host plants of D. noxia. Our results demonstrate the influence of host plant species/genotype and endophyte species/strain on expression of aphid resistance, provide an explanation of the high levels of D. noxia resistance in PI 314696 and PI 440420 previously reported in the literature, and underscore the potential importance of endophytic fungi in conferring insect resistance in wild barley.     

植物内生真菌抗肿瘤活性代谢产物研究进展

癌症已成为全球头号杀手,迫切需要从自然界寻找更新、更有效的抗肿瘤药物。植物内生真菌是指生活在宿主植物体内,不会对宿主植物组织引起明显病害症状的一类真菌。众多研究表明,植物内生真菌在寻找抗肿瘤药物中起着至关重要的作用。随着植物内生真菌研究的深入,从植物内生真菌中寻找新的抗肿瘤活性成分已成为研究的热点。大量的抗肿瘤活性成分从植物内生真菌中分离出来,并表现出良好的抗肿瘤活性。目前,植物内生真菌抗肿瘤活性代谢产物主要有紫杉醇、喜树碱、长春新碱,鬼臼毒素等等,本文主要对近年来植物内生真菌抗肿瘤活性成分的研究进展进行了综述。     

Psychrophilic endophytic fungi with biological activity inhabit Cupressaceae plant family

Psychrophilic microorganisms are cold-adapted organisms that have an optimum growth temperature below 15 °C, and often below 5 °C. Endophytic microorganisms live inside healthy plants and biosynthesize an array of secondary metabolites which confer major ecological benefits to their host. We provide information, for the first time, on an endophytic association between bioactive psychrophilic fungi and trees in Cupressaceae plant family living in temperate to cold, semi-arid habitats. We have recovered psychrophilic endophytic fungi (PEF) from healthy foliar tissues of Cupressus arizonica, Cupressus sempervirens and Thuja orientalis (Cupressaceae, Coniferales). In total, 23 such fungi were found out of 110 endophytic fungal isolates. They were identified as ascomycetous fungi, more specifically Phoma herbarum, Phoma sp. and Dothideomycetes spp., all from Dothideomycetes. The optimal growth temperature for all these 23 fungal isolates was 4 °C, and the PEF isolates were able to biosynthesize secondary metabolite at this temperature. Extracted metabolites from PEF showed significant antiproliferative/cytotoxic, antifungal and antibacterial effects against phytopathogenic fungi and bacteria. Of special interest was their antibacterial activity against the ice-nucleation active bacterium Pseudomonas syringae. Accordingly, we suggest that evergreen Cupressaceae plants may benefit from their psychrophilic endophytic fungi during cold stress. Whether such endosymbionts confer any ecological and evolutionary benefits to their host plants remains to be investigated in vivo.     

内生真菌及其对宿主植物生态适应性的影响

从植物-真菌的相互作用和植物体内微生态学的角度,综述近年来植物-内生真菌互作关系的研究进展,分析在生物和非生物环境因子胁迫下,植物内生真菌对宿主生态适应能力的影响及其可能的机制,特别强调将植物及其内生微生物作为一个整体研究的重要性,表明植物内生真菌作为一类重要的微生物资源,可以在增强植物的生态适应性,缓解全球气候变化对植物和人类的压力方面发挥其重要的作用,因而具有良好的应用前景。     

Potential analysis of grass endophytes Neotyphodium as biocontrol agents

Many grasses in the subfamily Pooideae develop symbioses with Neotyphodium fungal endophytes, which exist widely in nature. The stably symbiotic relationship not only ensures accessible nutrients required by Neotyphodium fungal endophytes, but also significantly increases the resistance of host grasses to biological stresses through the production of secondary metabolites. Previous studies show that infected grasses with endophytic fungi have prominently enhanced resistance to pests, plant diseases, companion plants and other biological stresses. Grass endophytic fungi show remarkable resistant to at least 79 species of pests from three classes; arachnida, nematode and insecta, and to at least 22 species of pathogenic fungi. Although the biotechnological application of endophytic fungi in grass breeding for variety selection and quality improvement has progressed well, opportunities remain for further exploring the use of fungal endophytes among different host grasses coupled with the examination of genetic stability of Neotyphodium in novel host grasses. In the future application of endophytic fungi as a bio-control method, researchers should not only consider specificities of host grasses, but also need to have comprehensive analysis and knowledge about the mutual relationships among grasses, endophytic fungi and ecological environments, which will help use endophytic fungi to better serve humanity.     

禾草内生真菌作为生防因子的潜力分析

早熟禾亚科多种禾草可与Neotyphodium内生真菌形成禾草-内生真菌共生体, 这种植物-微生物共生体性状较为稳定, 且在自然界中广泛存在。禾草-内生真菌共生体稳定的互利共生关系不但保证了内生真菌所需的全部营养物质, 而且共生体产生的次生代谢物又可显著提高宿主禾草对生物胁迫的抗逆性。众多研究表明, 内生真菌的侵染可显著提高宿主禾草对虫害、病害及伴生植物等多种生物胁迫的抗性。据不完全统计, 禾草内生真菌对蛛形纲、线虫纲、昆虫纲3个纲至少79个种的害虫表现出较明显的抗性, 对至少22个种的病原真菌表现出明显的抗性。尽管利用内生真菌进行禾草品种选育及其品质改良的技术日趋成熟, 但是内生真菌在不同宿主禾草之间高效的替代转化技术, 及其在宿主体内遗传的稳定性仍有待于进一步深入探索。研究者把禾草内生真菌作为生防手段, 在未来的应用过程中不应只考虑其与宿主禾草之间的共生特异性, 而应更全面地分析禾草-内生真菌-生态环境之间的相互关系, 让内生真菌更好地为人类服务。