Advances in research of Achnatherum inebrians-Epichloë endophyte symbionts

有关醉马草(Achnatherum inebrians)内生真菌(Epichloë gansuensis, E. inebrians)共生体的研究, 代表了我国禾草内生真菌研究领域的重要方向, 使中国的醉马草-内生真菌与美国的苇状羊茅(Festuca arundinacea)-内生真菌(E. coenophiana)和新西兰的多年生黑麦草(Lolium perenne)-内生真菌(E. festucae var. lolii)成为禾草内生真菌国际三大研究分支。该文综述了近30年来对醉马草内生真菌共生体的系统研究, 包括: 内生真菌的分布、带菌率、检测方法、多样性, 内生真菌提高宿主的抗旱、耐寒、耐盐碱、耐重金属、抗虫、抗病等抗逆性及其机理, 共生体产生的生物碱等次生代谢物, 对草食动物的毒性, 及其在草地生态系统中的作用等。研究者实验证实了醉马草本身无毒, 只有当内生真菌与醉马草共生并产生麦角新碱和麦角酰胺等麦角类生物碱后才能导致采食醉马草家畜中毒。文章展望了醉马草内生真菌基因组学和功能分析, 利用杀菌剂杀死内生真菌进行醉马草脱毒, 利用无毒内生真菌菌株进行饲用醉马草新品种选育, 利用有毒醉马草内生真菌共生体进行抗虫防鸟的机场绿化新品种选育及生物源农药与医药开发等。     

禾草内生真菌生物碱的研究进展

综述了国内外近20a以来在禾草内生真菌生物碱方面的研究进展。目前,已发现至少4大类10余种生物碱与内生真菌有关。各类生物碱中典型代表物的分子结构已完全清楚,部分内生真菌在离体条件下可产生除黑麦草碱外的生物碱,但产碱量较其在植物体中所产生的低很多,可相差150余倍之多。随着生物技术的发展,美国、新西兰等国在波胺、麦角碱和loline的生物合成途径方面已有了初步的进展,对个别具有重要功能基因以及其所编码的酶已有了深入研究。各种生物碱的致毒机理尚未完全清楚,除饱和吡咯化合物为新陈代谢类毒素和神经性毒素外,其余3类生物碱均为神经性毒素。生物碱可增加禾草对40余种害虫的抗性,并可增加对某些线虫和病害的抗性。诸多因素均可影响寄主植株中生物碱的种类和浓度,包括寄主植株：种群和生态型,植物品种和基因型,植株的部位和生长期;环境：气候因素,土壤养分,季节和年度变化;内生真菌菌株和草地管理利用方式等。用于生物碱检测的主要方法为预分离检测法和直接检测法,其中高效液相色谱法以其分离能力强、选择性高、测定灵敏度高,操作简单,可在室温下进行,应用范围极广的优点而广泛应用。目前,国际关于禾草内生真菌生物碱研究的重点包括创造不含对家畜有毒素的有益禾草-内生真菌共生体,开展基因工程研究以及合理利用生物碱,使其成为新一代的“生物农药”。     

禾草类内生真菌的研究进展*

感染内生真菌的禾草在牧草和草坪业上具有重要的生态和经济意义。关于禾草和内生真菌的互利共生作用已有大量报道,中就近年来有关禾草类内生真菌的起源和进化、内生真菌的生物和生态学作用以及内生真菌的应用前景方面的研究进展作一综述,以期更好地利用内生真菌造福人类。     

不同pH对醉马草内生真菌共生体幼苗生长和产碱的影响

该研究考察了不同pH(3、5、7、9和11)处理对醉马草内生真菌共生体幼苗的形态指标、叶绿素、可溶性糖以及麦角酰胺和麦角新碱含量的影响,以明确共生体幼苗生长和产碱的最适pH条件。结果表明：（1）醉马草内生真菌共生体幼苗株高、根长、分蘖和生物量等均在pH 7时（达到最大）显著高于其他4个酸碱处理,而且4个生长指标在强碱性处理下均显著高于强酸处理。（2）幼苗叶绿素含量也在pH 7的中性条件下（达到最高）显著高于pH 3、pH 5和pH 11处理,且强碱性处理下叶绿素含量也显著高于强酸处理,但幼苗可溶性糖含量在不同pH处理之间没有显著变化。（3）在相同时间内,醉马草内生真菌共生体幼苗麦角酰胺和麦角新碱含量均随环境pH值升高而增加;在处理时间为15 d时,麦角酰胺在pH 9~11处理下积累量最多,麦角新碱含量在pH 11处理下达到最大值,酸性条件抑制醉马草内生真菌共生体幼苗产碱。研究发现,醉马草内生真菌共生体幼苗在中性环境条件下生长最佳,且在碱性条件下生长较酸性条件下生长良好,而体内麦角酰胺和麦角新碱的积累在强碱性条件下达到峰值,可将醉马草内生真菌共生体应用于盐碱地的栽培绿化。     

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。     

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     

植物内生真菌防治根结线虫研究进展

根结线虫病是对农作物危害严重且难以防治的病害,并随着我国设施农业的发展日趋严重。常规的化学防治方法因毒性大、破坏生态环境而不适应农业的可持续发展。作为一种能稳定寄生在作物体内的生物防治真菌,内生真菌通过抑制卵的孵化、降低J2期线虫幼虫活力、抑制线虫的入侵、延缓雌虫发育、减少产卵数目、降低作物根中根结和卵块数量,来实现稳定、高效、安全地防治根结线虫病害。近年来,内生真菌的作用机制得到广泛关注和研究,取得了显著进展。本文综述近年来内生真菌生物防治根结线虫机制的研究进展,总结了内生真菌直接攻击、资源竞争、代谢物胁迫、防御激活等4种主要机制,探讨其存在的问题,以期为进一步开发、应用植物内生真菌进行生物防治提供帮助。     

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。     

光照强度和光周期对醉马草内生真菌共生体幼苗生长和生物碱积累的影响

该研究通过测定不同光周期(8、12、14和16h·d~(-1))和光照强度(1 500、2 500和3 500lux)处理下醉马草内生真菌共生体幼苗的形态指标以及叶绿素、可溶性糖、麦角酰胺和麦角新碱的含量,以明确醉马草内生真菌共生体幼苗生长和生物碱积累的最适光照条件,为醉马草内生真菌共生体的应用提供理论依据。结果发现:(1)醉马草内生真菌共生体幼苗株高、叶绿素和可溶性糖含量随着光照时间的延长而增加,并在光周期为16h·d~(-1)时显著(P0.05)高于其他处理;根长、单株分蘖数和单株生物量均先升高后降低,且均在14h·d~(-1)处理下达到最大值。(2)随着光照强度的增加,醉马草内生真菌共生体幼苗4个形态指标以及叶绿素和可溶性糖含量均呈增加的趋势,并在光照强度为3 500lux时均达到最大值。(3)随着处理时间的延长,醉马草内生真菌共生体幼苗麦角酰胺和麦角新碱的含量在不同光周期处理下均呈上升的趋势,并在光周期为12h·d~(-1)和光照强度为2 500lux处理下麦角酰胺和麦角新碱的含量均显著高于其他处理(P0.05),且在处理第15天时达到最大值。研究表明,适宜的光照环境能显著促进醉马草内生真菌共生体幼苗生长和生物碱的积累,并以12h·d~(-1)光周期和2 500lux光照处理下最有利于麦角酰胺和的麦角新碱积累。     

Effects of endophyte infection on the competitive ability of Achnatherum sibiricum depend on endophyte species and nitrogen availability

Aims The leaves of almost all terrestrial plant species are colonized by endophytic fungi. Compared to agronomic grasses, which usually harbor few endophytes, native grasses generally possess greater endophyte species diversity. Existing studies examining endophyte effects on natural grasses under competition normally considered the infection status (infected or uninfected), and rarely considered endophyte species.     

内生真菌感染对宿主羽茅及邻生植物抗病性的影响

Epichloë内生真菌能够影响宿主植物的抗病性, 但目前关于内生真菌感染对宿主邻生植物抗病性影响的研究很少。该研究首先调查了内蒙古呼伦贝尔天然草地中植物病害发生情况, 发现在常见牧草中禾本科植物病害最为严重, 其中感染内生真菌的羽茅(Achnatherum sibiricum)的总病情指数最低。进一步比较了与羽茅邻生及非邻生植物的病害差异, 结果显示羽茅显著降低了邻生羊草(Leymus chinensis)、狼针草(Stipa baicalensis)和冰草(Agropyron cristatum)的褐斑病病情指数。为探究羽茅对邻生植物病害的影响是否与其内生真菌有关, 分别从内生真菌发酵液、离体叶片和植株层面进行了室内验证试验。结果表明: 羽茅的2种内生真菌Epichloë gansuensis和Epichloë sibiricum发酵液显著抑制了新月弯孢(Curvularia lunata)、根腐离蠕孢(Bipolaris sorokiniana)、核盘菌(Sclerotinia sclerotioru)和三叶草核盘菌(Sclerotinia trifoliorum)的生长; 对于离体叶片, E. gansuensis和E. sibiricum显著减小了宿主叶片受4种病原真菌侵染后的病斑长度。在新月弯孢和根腐离蠕孢的植株接种条件下, 内生真菌显著降低了宿主羽茅及邻生植物羊草的病害程度。该研究首次发现并验证了羽茅内生真菌对邻生禾草抵御褐斑病有一定的增益作用。     

Advances in research of Achnatherum inebrians-Epichloë endophyte symbionts北大核心CSCD

Research on drunken horse grass (Achnatherum inebrians)-endophytic fungi (Epichloë gansuensis, E. inebrians) represented an important area of endophyte research in China. Compared with tall fescue (Festuca arundinacea)-endophyte (E. coenophiala) in the United States and perennial ryegrass (Lolium perenne)-endophyte (E. festucae var. lolii) in New Zealand, A. inebrians-endophyte research in China is becoming a leading branch of endophyte research. The studies of A. inebrians-endophyte symbionts over the past three decades were reviewed, mainly including endophyte distribution, detection methods, infection rate, fungal diversity, stress tolerances of drought, cold, salt, heavy metal, pest, disease and their mechanisms, secondary metabolite of alkaloids, feeding experiments and its ecological functions etc. Research reveals that drunken horse grass itself has no toxicity, the grazed animals will be intoxicated if the grass bears infection by Epichloë endophytes and produces ergot alkaloids. Future research will focus on E. gansuensis and E. inebrians genomics and endophyte functions. In other research we will create new grass cultivars that are 1) sanitized of the endophyte and non-toxic, 2) utilizing novel endophytes that deter birds feeding and 3) utilizing endophytes that are modified to produce novel fungicides, pesticides or medicines. © 2018 Editorial Office of Chinese Journal of Plant Ecology. All rights reserved.     

Relationships between the genetic distance of Epichloë festucae isolates and the ergovaline and peramine contents of their Festuca rubra hosts

In semiarid grasslands of western Spain, plants of Festuca rubra are frequently infected by Epichloë festucae , an endophyte capable of producing the alkaloids ergovaline, toxic to mammals, and peramine, toxic to insects. The objectives of this paper were to estimate the variability in the content of ergovaline and peramine in plants of Festuca rubra from natural populations and to determine the relationship between the genetic distance among Epichloë strains and the alkaloid contents of plants infected by them. The results showed that a significant variation exits in the ergovaline and peramine contents of infected F. rubra plants, and that variation patterns are different for each alkaloid. Ergovaline content showed a significant variation between years. In contrast, most variation observed in the concentration of peramine occurred between populations. Mantel tests, constrained correspondence analysis (CCA) and generalised analysis of molecular variance (GAMOVA) all revealed a significant relationship between the genetic distances among 35 Epichloë isolates, and the differences in peramine content of their host plants. The relationship observed for ergovaline was more ambiguous. This indicates that plants infected by genetically close isolates of Epichloë festucae tend to be similar in terms of peramine content, while the ergovaline content of infected plants seems to be more dependent on external factors. The results of this study suggest that the selection of Epichloë festucae endophytes based on the alkaloid content of their host plants can be a successful strategy to select good or bad producers of peramine, but may not be a reliable method to detect fungal strains associated with particular ergovaline production profiles.     

Foliar fungal endophyte community structure is independent of phylogenetic relatedness in an Asteraceae common garden

Phylogenetic distance among host species represents a proxy for host traits that act as biotic filters to shape host‐associated microbiome community structure. However, teasing apart potential biotic assembly mechanisms, such as host specificity or local species interactions, from abiotic factors, such as environmental specificity or dispersal barriers, in hyperdiverse, horizontally transmitted microbiomes remains a challenge. In this study, we tested whether host phylogenetic relatedness among 18 native Asteraceae plant species and spatial distance between replicated plots in a common garden affects foliar fungal endophyte (FFE) community structure. We found that FFE community structure varied significantly among host species, as well as host tribes, but not among host subfamilies. However, FFE community dissimilarity between host individuals was not significantly correlated with phylogenetic distance between host species. There was a significant effect of spatial distance among host individuals on FFE community dissimilarity within the common garden. The significant differences in FFE community structure among host species, but lack of a significant host phylogenetic effect, suggest functional differences among host species not accounted for by host phylogenetic distance, such as metabolic traits or phenology, may drive FFE community dissimilarity. Overall, our results indicate that host species identity and the spatial distance between plants can determine the similarity of their microbiomes, even across a single experimental field, but that host phylogeny is not closely tied to FFE community divergence in native Asteraceae.     

真菌性鼻-鼻窦炎研究进展

真菌性鼻-鼻窦炎是鼻科临床常见的一种特异性感染性疾病。传统观点认为,真菌性鼻-鼻窦炎(fungalrhino-sinusitis,FRS)多在机体长期使用抗生素、糖皮质激素、免疫抑制剂及接受放射治疗等情况下发生,也可在一些慢性消耗性疾病如糖尿病、烧伤致机体抵抗力下降时发生。但近年来在健康体检中也有发现FRS,造成这种情况的原因目前还不太清楚,有待进一步研究。由于引起致病的病原真菌种类不同,真菌性鼻窦炎的临床类型、诊断、治疗及临床疗效果等均有各自的特点,以下针对病因、临床类型及表现、诊断、治疗及预后等方面进行回顾。     

Infection with the fungal endophyte Epichloë festucae may alter the allelopathic potential of red fescue

Red fescue (Festuca rubra) is a perennial grass used as both forage and turfgrass. Asymptomatic plants of this species are systemically infected by the fungal endophyte Epichloë festucae, which has a beneficial effect on the infected plants. The aim of this study was to determine the effect of the endophyte Epichloë festucae on the allelopathic potential of F. rubra against four associated pasture species that are also considered as weeds in lawns, Trifolium pratense, Trifolium repens, Lotus corniculatus and Plantago lanceolata. Two experiments were designed to evaluate the allelopathic effect of extracts from the roots and leaves of endophyte‐infected (E+) and non‐infected (E?) plants on the germination and seedling growth of the four target species. Regardless of the endophyte status of the host plant, leaf extracts elicited a stronger reduction in germination and seedling growth than root extracts. Extracts from E+ plants reduced the speed of germination index of Trifolium spp. to a greater extent than those from E? plants. Radicle length of the target species was the parameter most affected by the presence of the endophyte in F. rubra. Root extracts from E+ plants had a greater inhibitory effect on the radicle growth of the target species than did root extracts from E? plants. A greater concentration in total phenolic compounds was found in the roots of E+ plants than of E?; however, this difference was not observed in the leaves. Thus, the allelopathic potential of F. rubra is altered in infected plants.