Can seed-borne endophytes promote grass invasion by reducing host dependence on mycorrhizas?

Symbiotic interactions between plants and microorganisms have recently become the focus of research on biological invasions. However, the interaction between different symbionts and their consequences in host-plant invasion have been seldom explored. Here, we propose that vertically transmitted fungal endophytes could reduce the dependency of invasive grasses on mycorrhizal fungi allowing host establishment in those environments where the specific mutualist may be not present. Through analyzing published studies on nine grass species, we evaluated the effect of seed-borne Epichloë endophytes on the relationship of invasive and non-invasive grasses with arbuscular mycorrhizal fungi (AMF), a symbiosis known to be fundamental for plant fitness and invasion success. The endophyte effect on AMF colonization differed between invasive and non-invasive grasses, reducing mycorrhization only on invasive species but with no impact on their biomass. These results allowed us to propose that Epichloë endophytes could reduce the dependency of host plants on the mutualism with AMF, promoting host grass establishment and subsequent invasion. Simultaneous interactions with different types of mutualists may have profound effects on the host-plant fitness facilitating its range expansion. Our findings suggest that some specific mutualistic fungi such as epichloid endophytes facilitate host invasion by reducing the requirements of the benefits derived from other mutualisms.     

禾本科植物内生真菌资源及其物种多样性

植物内生真菌是当代微生物资源研究的一个热门话题,对禾本科植物内生真菌资源研究进行了总结。与冷季型禾本科植物共生的麦角菌科epichlo内生真菌是目前的研究重点,包括偶尔在宿主植物体表形成子座的Epichlo属真菌和几乎完全不形成症状的Neotyphodium属真菌。具体介绍该真菌类群的物种多样性及其地理分布多样性,概括了它们在全球各大区域的分布特征。对近年来中国迅速发展的epichlo内生真菌的研究作了总结,最后展望了国际国内epichlo内生真菌资源探索的发展方向。特别指出:冷季型禾本科植物内生真菌在南美洲和亚洲,暖季型禾本科植物内生真菌在热带亚热带地区的研究还相对薄弱,值得今后继续加强探索和挖掘。     

Geographic distribution patterns of vertically transmitted endophytes in two native grasses in Argentina

The incidence of epichloid endophytes in populations of wild grasses is usually variable, and the knowledge about distribution patterns and how environmental factors affect such an incidence is limited. Here we performed a broad scale survey data to study whether the distribution patterns and the incidence of vertically-transmitted endophytes in populations of two native grasses from South-America, Poa lanuginosa Poir. and Poa bonariensis (Lam.) Kunth., are associated with environmental characteristics. We also characterized the endophytes from different populations to establish if the genotype of the endophytes is also correlated with environmental variables. The incidence of endophytes ranged from 0 to 100 % in both host species. In P. lanuginosa, endophytes were only found in populations on sandy coastal dunes and their incidence was positively associated with winter regime rainfall and soil water availability in the growing season. In P. bonariensis, endophytes were only found in populations in xerophytic forests and their incidence was highly associated with plant community. The distributions of infested populations suggested that the endophytes are not found in those areas with the most favorable or most stressing growth conditions accordingly to climatic or edaphical characteristics. Only the vertically transmitted hybrid endophyte species Neotyphodium tembladerae was detected in both host species. Under the hypothesis of vertical transmission, these results suggested that the endophyte should have been lost in endophyte free populations but is maintained in populations established in environments presenting moderate stress as salinity or short drought periods.     

Interactions among fungal endophytes,grasses and herbivores

The interaction between two species often depends on the presence or absence of a third species. One widespread three-species interaction involves fungal endophytes infecting grasses and the herbivores that feed upon them. The endophytes are allied with the fungal family Clavicipitaceae and grow systemically in intercellular spaces in above-ground plant tissues including seeds. Like relatedClaviceps species, the endophytes produce a variety of alkaloids that make the host plants toxic or distasteful to herbivores. A large number of grass species are infected, especially cool-season grasses in temperate areas. Field and laboratory studies have shown that herbivores avoid infected plants in choice trials and suffer increased mortality and decreased growth on infected grasses in feeding experiments. Resistance to herbivores may provide a selective advantage to infected plants in competitive interactions with noninfected plants. Recent studies have shown that differential herbivory can reverse competitive hierarchies among plant species. Both endophyte-infected and noninfected tall fescue grass (Festuca arundinacea) are outcompeted by orchardgrass (Dactylis glomerata) in the absence of insect herbivory. However, when herbivores are present infected tall fescue outcompetes orchardgrass. These results suggest that the frequency of infection in grass species and grassland communities will increase over time. Several studies are reviewed illustrating increases in infection frequency within grass populations subject to herbivore pressure. Endophytic fungi may be important regulators of plant-herbivore interactions and so indirectly affect the structure and dynamics of plant communities.     

禾草-内生真菌共生体在草地农业系统中的作用

综述了国内外近年对禾草 -内生真菌 (N eotyphodium)的研究进展。全世界现已报道 14种禾草内生真菌 ,与 2 3个属的禾草形成共生体 ,我国已在 13属 2 5种天然草地禾草中发现内生真菌 ,其中发草属 (Descampsia)、大麦属 (H ord eum)和赖草属(L eymus)以往在国际文献中未曾报道。内生真菌在禾草体内产生的生物碱 ,致使采食带菌禾草的马、牛、羊、鹿等家畜产生中毒症状 ,每年给美国、新西兰等国造成的经济损失达 6.4亿美元之多 ,对其毒理研究取得了进展。已发现我国醉马草 (Achnatheruminebrians)对家畜的毒性与内生真菌的侵染有关。内生真菌侵染增加可使禾草对 42种害虫的抗性 ,并可增加对某些线虫和病害的抗性。与不带菌禾草相比 ,带菌禾草的另一特点是抗逆性强 ,牧草产量高。国际在该领域的研究主要集中在多年生黑麦草(L olium p erenne)和高羊茅 (Festuca arundinacea)。我国的研究发现 ,带菌布顿大麦草 (H ordeum bodg anii)和圆柱披碱草(Elymus cylind ricus)的牧草产量分别增加 3 3 .3 %和 2 78.8% ,分孽数分别增加 13 6.8%和 84.5%。目前 ,国际研究的重点包括大规模开展内生真菌生物学与生态学特性的研究 ,创造不含对家畜有害毒素的有益禾草 -内生真菌共生体 ,培育带内生真菌的草坪草品种 ,培育抗毒     

禾草内生真菌对其它微生物的影响研究进展

禾草内生真菌在宿主植物的茎叶等地上组织中普遍存在,不仅能够提高禾草对生物与非生物逆境的抗性,而且能够对周围环境中的不同微生物类群产生影响。主要总结了禾草Neotyphodium/Epichlo内生真菌对病原真菌、丛枝菌根真菌和土壤微生物的影响及其作用机理。发现禾草内生真菌普遍存在对病原真菌的抑制作用,而对丛枝菌根真菌存在不对称的竞争作用,且因种类而异。禾草内生真菌对土壤微生物群落的作用则会随着土壤类型和时间等外界因素发生变化。禾草内生真菌对不同类群微生物的影响机制主要包括:通过生态位竞争、抑菌物质分泌、诱导抗病性等对病原真菌造成影响;通过根系化学物质释放、营养元素调节、侵染条件差异等对丛枝菌根真菌造成影响;通过根际沉积物和凋落物等对土壤微生物群落造成影响。禾草内生真菌产生的生物碱能提高宿主植物对包括昆虫在内草食动物采食的抗性,影响病原菌的侵入、定殖和扩展;根组织分泌物中包含次生代谢产物能够抑制菌根真菌、土传病原真菌及其它土壤微生物的侵染与群落组成;也可能通过次生代谢物影响禾草的其它抗性。因此,禾草内生真菌在植物-微生物系统中的作用应该给予更多的关注和深入研究。     

Evolutionary origins and ecological consequences of endophyte symbiosis with grasses

Over the past 20 yr much has been learned about a unique symbiotic interaction between fungal endophytes and grasses. The fungi (Clavicipitaceae, Ascomycota) grow intercellularly and systemically in aboveground plant parts. Vertically transmitted asexual endophytes forming asymptomatic infections of cool-season grasses have been repeatedly derived from sexual species that abort host inflorescences. The phylogenetic distribution of seed-transmitted endophytes is strongly suggestive of cocladogenesis with their hosts. Molecular evidence indicates that many seed-transmitted endophytes are interspecific hybrids. Superinfection may result in hyphal fusion and parasexual recombination. Most endophytes produce one or more alkaloid classes that likely play some role in defending the host plant against pests. Hybridization may have led to the proliferation of alkaloid-production genes among asexual endophytes, favoring hybrids. The ergot alkaloid ergovaline, lolitrems, and lolines are produced by only a single sexual species, Epichlo? festucae, but they are common in seed-transmitted endophytes, suggesting that E. festucae contributed genes for their synthesis. Asexual hybrids may also be favored by the counteracting of the accumulation of deleterious mutations (Muller's rachet). Endophyte infection can provide other benefits, such as enhanced drought tolerance, photosynthetic rate, and growth. Estimates of infection frequency have revealed variable levels of infection with especially high prevalence in the subfamily Pooideae. Longitudinal studies suggest that the prevalence of seed-transmitted endophytes can increase rapidly over time. In field experiments, infected tall fescue suppressed other grasses and forbs relative to uninfected fescue and supported lower consumer populations. Unlike other widespread plant/microbial symbioses based on the acquisition of mineral resources, grass/endophyte associations are based primarily on protection of the host from biotic and abiotic stresses.     

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.     

Diversity and distribution of Neotyphodium-infected grasses in Argentina

In the present work, we studied the presence of “epichlo?/neotyphodium endophytes” in native grasses from Argentina. An extensive area of this country, representative of several different environments, was studied. Nine new host species of the genera Briza, Bromus and Poa were observed to be infected with asexual seed-borne endophytes. Epichlo? stromata were not observed on any grass species. The incidence of infection in natural populations was highly variable among host species and among populations of the same host species. Morphological characterization revealed differences among the endophytes of different host species and among endophytes of different populations of the same host species. We also summarize the previous knowledge and present unpublished data on host diversity and distribution of these fungi in Argentina. Our results are discussed and compared with previous studies on endophytes in Argentina. This work supports the hypothesis that sexual species (Epichlo?) are not present in the southern hemisphere, and suggests the existence of a high diversity of asexual endophytes in South America.     

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。     

Variable effects of endophytic fungus on seedling establishment of fine fescues

Seedborne systemic endophytic fungi of grasses are thought to be plant mutualists, because they have been shown to improve their host’s resistance against biotic and abiotic stresses. The interactions in plant–endophyte associations vary from mutualistic to parasitic with environmental conditions and the genotypes of interacting species. The possible pros and cons of endophytic fungi are expected to be most evident during the seedling establishment, where host fitness is most directly affected. If this holds true, endophytes may play a focal role in local adaptation of hosts to different environments. We examined if endophyte-infected and uninfected seeds and seedlings of two native grass species, Festuca rubra and F. ovina, differ in seed germination and seedling growth rates under greenhouse conditions. The germination of F. rubra seeds was also studied in the field. This is the first time that the effects of Epichloë endophyte on seedling establishment of fine fescues from natural populations have been experimentally evaluated. Mother plant (seed family) had a marked effect on many response variables in both grass species. Length and mean biomass of tillers of endophyte-infected (E+) F. ovina seedlings were lower, but root:shoot ratios were higher than in endophyte-free (E?) seedlings. In F. rubra, the effects of the endophyte were dependent on the habitat where the seeds were collected. The E+ seeds from river banks germinated faster than E+ seeds from meadows, and E+ seedlings from the river banks produced fewer but taller and heavier tillers than the other seedlings. Our data suggest that the effects of the endophyte infection on the seedling stage of fine fescues are dependent the species of grass, host genetic background and mother plant habitat. The germination strategy and growth form of E+ red fescue seedlings from river banks may be beneficial to surviving in the harsh conditions of that habitat.     

Mutualistic asexual endophytes in a native grass are usually parasitic

Asexual systemic fungi that live symbiotically within grasses are viewed as strong mutualists on the basis of theory and empirical studies of introduced agronomic grasses. Evolutionary theory predicts that microbial symbionts that lose sexuality and rely on propagules of their hosts for transmission should evolve to benefit their hosts. Fungal endophytes of some cultivated turf and pasture grasses are well known for increasing plant performance and competitive abilities, especially under stress, and increasing resistance to herbivores, pathogens, and root-feeders by virtue of fungal alkaloids. The assumption of mutualism, however, has rarely been tested in native grasses, which often harbor high but variable frequencies of systemic asexual endophytes. We tested the effect of Neotyphodium infections for the native grass Arizona fescue in a 3-yr field experiment. We strictly controlled host genotype and manipulated soil moisture and nutrients. Infection generally decreased host growth in terms of plant volume, number of tillers, and dry mass of shoots and roots. Infected plants also showed decreased reproduction in terms of number and mass of seeds, and the seeds produced by infected plants had lower germination success than plants without their endophytes, suggesting that the negative effects of the symbiont are transferred to the next generation. Plant genotype strongly influenced host's growth and reproduction and interacted with the presence of the endophyte, but the interaction was usually in the direction of negative effects. Our results challenge the notion that systemic asexual endophytes must be plant mutualists for infections to persist in nature. We propose other hypotheses to explain the variable but usually high endophyte frequencies in natural populations of grasses.     

Epichlo? endophytes grow by intercalary hyphal extension in elongating grass leaves.

A fundamental hallmark of fungal growth is that vegetative hyphae grow exclusively by extension at the hyphal tip. However, this model of apical growth is incompatible with endophyte colonization of grasses by the symbiotic Neotyphodium and Epichlo? species. These fungi are transmitted through host seed, and colonize aerial tissues that develop from infected shoot apical meristems of the seedling and tillers. We present evidence that vegetative hyphae of Neotyphodium and Epichlo? species infect grass leaves via a novel mechanism of growth, intercalary division and extension. Hyphae are attached to enlarging host cells, and cumulative growth along the length of the filament enables the fungus to extend at the same rate as the host. This is the first evidence of intercalary growth in fungi and directly challenges the centuries-old model that fungi grow exclusively at hyphal tips. A new model describing the colonization of grasses by clavicipitaceous endophytes is described.     

Epichloë endophytes grow by intercalary hyphal extension in elongating grass leaves

A fundamental hallmark of fungal growth is that vegetative hyphae grow exclusively by extension at the hyphal tip. However, this model of apical growth is incompatible with endophyte colonization of grasses by the symbiotic Neotyphodium and Epichlo? species. These fungi are transmitted through host seed, and colonize aerial tissues that develop from infected shoot apical meristems of the seedling and tillers. We present evidence that vegetative hyphae of Neotyphodium and Epichlo? species infect grass leaves via a novel mechanism of growth, intercalary division and extension. Hyphae are attached to enlarging host cells, and cumulative growth along the length of the filament enables the fungus to extend at the same rate as the host. This is the first evidence of intercalary growth in fungi and directly challenges the centuries-old model that fungi grow exclusively at hyphal tips. A new model describing the colonization of grasses by clavicipitaceous endophytes is described.     

An ecological framework for understanding the roles of Epichloë endophytes on plant defenses against fungal diseases

Plants harbor a wide diversity of microorganisms in their tissues. Some of them have a long co-evolutionary history with their hosts, likely playing a pivotal role in regulating the plant interaction with other microbes such as pathogens. Some cool-season grasses are symbiotic with Epichloë fungal endophytes that grow symptomless and systemically in aboveground tissues. Among the many benefits that have been ascribed to endophytes, their role in mediating plant interactions with pathogens has been scarcely developed. Here, we explored the effects of Epichloë fungal endophytes on the interaction of host grasses with fungal pathogens. We made a meta-analysis that covered a total of 18 host grass species, 11 fungal endophyte species, and 22 fungal pathogen species. We observed endophyte-mediated negative effects on pathogens in vitro and in planta. Endophyte negative effects on pathogens were apparent not only in laboratory but also in greenhouse and field experiments. Epichloë fungal endophytes had negative effects on pathogen growth and spores' germination. On living plants, endophytes reduced both severity and incidence of the disease as well as colonization and subsequent infection of seeds. Symbiosis with endophytes showed an inhibitory effect on debilitator and killer pathogens, but not on castrators, and this effect did not differ among biotrophic or necrotrophic lifestyles. We found that this protection can be direct through the production of fungistatic compounds, the competition for a common resource, or the induction of plant defenses, and indirect associated with endophyte-generated changes in the abiotic or the biotic environment. Several mechanisms operate simultaneously and contribute differentially to the reduction of disease within grass populations.     

Do the costs and benefits of fungal endophyte symbiosis vary with light availability?

? Here, we examined whether fungal endophytes modulated host plant responses to light availability. First, we conducted a literature review to evaluate whether natural frequencies of endophyte symbiosis in grasses from shaded habitats were higher than frequencies in grasses occupying more diverse light environments. Then, in a glasshouse experiment, we assessed how four levels of light and the presence of endophyte symbioses affected the growth of six grass species. ? In our literature survey, endophytes were more commonly present in grasses restricted to shaded habitats than in grasses from diverse light environments. ? In the glasshouse, endophyte symbioses did not mediate plant growth in response to light availability. However, in the host grass, Agrostis perennans, symbiotic plants produced 53% more inflorescences than nonsymbiotic plants at the highest level of shade. In addition, under high shade, symbiotic Poa autumnalis invested more in specific leaf area than symbiont-free plants. Finally, shade increased the density of the endophyte in leaf tissues across all six grass species. ? Our results highlight the potential for symbiosis to alter the plasticity of host physiological traits, demonstrate a novel benefit of endophyte symbiosis under shade stress for one host species, and show a positive association between shade-restricted grass species and fungal endophytes.     

内生真菌对草坪植物病原真菌抑制作用的比较

分别从野生牧草羽茅(Achnatherum sibiricum (L.) Keng)、栽培种高羊茅(Festuca arundinacea Schreb.)(品种Millennium)、栽培种黑麦草(Lolium perenne L.) (品种Justus)中分离出内生真菌Neotyphodium sp.、N. coenophialum和N. lolli,通过体外培养法比较了这3种内生真菌对草坪植物病原真菌的抑制作用.结果表明,从羽茅中分离的内生真菌Neotyphodium sp.在两菌相交前对所有供试的病原真菌都有一定的抑制作用,其中对枝孢霉属(Cladosporium sp.)、弯孢霉属(Curvularia sp.)和拟茎点属(Phomopsis sp.)病原真菌的抑制效果尤为显著,对峙培养3d后的抑菌率分别达70.1%、52.3%和30.9%,营养竞争作用、重寄生作用是其主要的拮抗机制;从高羊茅中分离的内生真菌N. coenophialum对枝孢霉属病原真菌存在一定的抑制作用;而从黑麦草中分离的内生真菌N. lolli与病原真菌对峙培养时,病原真菌菌落慢慢侵占整个营养空间,内生真菌停止生长并逐渐褐变死亡.体外培养结果说明Neotyphodium sp.对供试病原真菌的拮抗效果优于N. coenophialum和N. lolli,由此推测Neotyphodium sp.与宿主植物羽茅的共生可能有利于宿主植物抵抗病原真菌的侵扰.     

禾草内生真菌的遗传多样性及其共生关系

 近百年的禾草内生真菌研究历经了由浅入深的过程,从最初的家畜中毒事件认识到是一种共生内生真菌存在的缘故,到如今利用分子生物学技术揭示其共生机制,人类发现这类植物内生真菌并非想象中的对生态系统无足轻重。Epichloë及其无性型Neotyphodium与禾本科植物是系统发生的互利共生关系,尤其是Neotyphodium可提高宿主抵抗环境胁迫的能力和抵御动物的取食,增强植物的竞争力。禾草内生真菌有3种生活史：有性生活史、无性生活史和兼性生活史,后者表明真菌在不同的宿主及环境下既能营有性生殖也可营无性繁殖,是一种更灵活而有效的生活史对策。对内生真菌分子系统学、生活史以及与宿主禾草协同进化的研究发现,Neotyphodium起源于禾草致病真菌Epichloë的某些种,或是Epichloë与Neotyphodium的种间杂交后代。植物和内生真菌各异的生活史策略,真菌的种间杂交,两者的协同进化亦或种群间基因流的差异,都促成了共生体多样化的基因组合（Genetic combination ）,也是其共生关系多样化的根源。内生真菌对宿主的有益作用只在特定基因型真菌、宿主和一定环境条件下才起作用,自然生态系统的共生关系要比农业系统复杂得多,是一个从互利共生至寄生关系的连续系统。未来对于更多共生体的遗传背景和基因与环境相互作用的阐明将有助于对禾草内生真菌共生关系本质更加深入的认识。     

Forest habitat characteristics affect balance between sexual reproduction and clonal propagation of the ectomycorrhizal mushroom Hebeloma cylindrosporum

Endophytic fungi, especially asexual, systemic endophytes in grasses, are generally viewed as plant mutualists, mainly through the action of mycotoxins, such as alkaloids in infected grasses, which protect the host plant from herbivores. Most of the evidence for the defensive mutualism concept is derived from studies of agronomic grass cultivars, which may be atypical of many endophyte-host interactions. I argue that endophytes in native plants, even asexual, seed-borne ones, rarely act as defensive mutualists. In contrast to domesticated grasses where infection frequencies of highly toxic plants often approach 100%, natural grass populations are usually mosaics of uninfected and infected plants. The latter, however, usually vary enormously in alkaloid levels, from none to levels that may affect herbivores. This variation may result from diverse endophyte and host genotypic combinations that are maintained by changing selective pressures, such as competition, herbivory and abiotic factors. Other processes, such as spatial structuring of host populations and endophytes that act as reproductive parasites of their hosts, may maintain infection levels of seed-borne endophytes in natural populations, without the endophyte acting as a mutualist.     

内生真菌对羽茅生长及光合特性的影响

通过温室栽培实验,以感染两种内生真菌(Neotyphodium sibiricum和Neotyphodium gansuence)和未感染内生真菌的羽茅(Achnatherum sibiricum)为实验材料,分析感染不同种内生真菌对宿主植物的生长及光合特性的影响。结果表明,感染两种内生真菌羽茅的株高和CO₂补偿点显著低于未染菌的羽茅,而染菌羽茅的蒸腾速率和气孔导度显著高于未染菌羽茅,但对于感染不同种内生真菌的羽茅,无论是分蘖数与生物量的积累还是光合生理值之间均无显著差异。