刈割干扰和养分添加条件下Epichloë内生真菌感染对羽茅所在群落多样性和生产力的影响

Epichloë内生真菌感染能够影响宿主植物的种内和种间竞争力, 但目前关于内生真菌感染对宿主植物所在群落多样性和生产力影响的研究较少。该研究以感染(E+)和不感染(E-)内生真菌的羽茅(Achnatherum sibiricum)及其原生境中常见的5种植物构建人工群落, 探究在不同养分水平和刈割条件下内生真菌对宿主植物群落的影响。结果表明: 内生真菌对宿主植物群落多样性的影响同刈割与否有关。在无刈割条件下, 内生真菌感染对宿主群落多样性无显著影响; 在刈割条件下, 内生真菌感染显著增加了宿主群落的多样性, 其原因在于内生真菌感染显著增加了群落中多度较小的冰草(Agropyron cristatum)和大针茅(Stipa grandis)等的多度, 而显著降低了优势种羊草(Leymus chinensis)的多度。内生真菌感染对群落生产力未见显著影响。研究发现养分添加对内生真菌作用的影响只出现在宿主植物水平, 而对宿主植物所在群落未产生显著影响, 内生真菌对宿主羽茅的促进作用只出现在养分添加条件下。     

内生真菌感染对宿主植物高羊茅锌耐受性的影响

以感染内生真菌(Neotyphodium coenophialum)和未感染内生真菌的高羊茅(Festuca arundinacea Schreb.)为实验材料,在营养液中加入ZnSO4进行锌胁迫实验,分析内生真菌对宿主植物锌耐受性的影响。与未感染内生真菌的植株相比,内生真菌感染对高羊茅的总生物量没有显著增益作用,但增加了分蘖数和叶片延伸生长累积值。内生真菌感染降低了高羊茅中Zn2+的总含量,改变了锌在高羊茅中的分配,增加叶鞘中锌的含量,减少叶片中锌的含量。在高锌浓度下,内生真菌感染对净光合速率的变化没有影响,但是显著提高了其宿主的PSⅡ光化学效率(Fv/Fm)。总体来看,内生真菌感染改善宿主高羊茅的锌耐受性。     

感染羽茅的香柱菌属内生真菌对丛枝菌根真菌孢子萌发的影响

内生真菌与丛枝菌根(AM)真菌是构成草原生态系统的重要组成部分.内生真菌会抑制其宿主植物的AM真菌侵染率.本研究以感染2种香柱菌属内生真菌[Epichloё gansuensis(Eg)和E. sibirica(Es)]的天然禾草羽茅为供试材料,进行体外纯培养的内生真菌培养滤液、感染内生真菌的羽茅叶片(包括鲜叶和枯叶)浸提液,以及根系分泌物对摩西球囊霉(Gm)和幼套球囊霉(Ge)2种AM真菌孢子萌发影响试验.结果表明: 香柱菌属内生真菌的培养滤液会显著抑制2种AM真菌孢子的萌发,而感染香柱菌属内生真菌的羽茅根系分泌物只对Ge孢子萌发有显著抑制作用,且上述抑制作用与内生真菌种类无关;鲜叶浸提液对Gm和Ge的孢子萌发率均无显著影响,而枯叶浸提液对Ge的孢子萌发有显著抑制作用.在自然生态系统中,香柱菌属内生真菌通常存在于宿主植物体内,可能通过影响宿主植物的根系分泌物来影响AM真菌孢子的萌发.     

内生真菌感染对黑麦草抗盐性的影响

以感染内生真菌的多年生黑麦草（Lolium perenne L．）（SR4000）为实验材料,建植内生真菌感染（EI）和不感染（EF）的黑麦草种群,并对其进行盐胁迫实验,通过观察生长和生理生态指标的变化,分析内生真菌对宿主植物抗盐性的影响。结果表明,内生真菌感染对宿主黑麦草的营养生长没有增益效应,相反在高盐浓度下,EI种群的分蘖能力和地上部分生物量均低于EF种群;但内生真菌能够改变宿主种群生物量的分配格局,将更大比例的生物量分配于根系。在高盐浓度下,内生真菌感染可导致黑麦草叶内的脯氨酸含量显著增加、可溶性糖含量显著降低,但对PSⅡ光化学效率Fv/Fm值的变化没有影响。总体来看,内生真菌感染并未改善宿主黑麦草的抗盐性。     

不同浓度和形态磷处理下内生真菌感染对高羊茅的影响

以感染内生真菌(endophyte-infected,EI)和不感染内生真菌(endophyte-free,EF)的高羊茅(Festuca arundinacea Schreb.)为材料,在温室沙培条件下研究内生真菌对高羊茅适应缺磷及利用不同形态磷肥的影响。结果表明,1)缺磷条件下,高羊茅EI和EF植株生长差异不显著;正常供磷条件下,高羊茅EI植株拥有更多分蘖数和绿叶数。说明正常供磷条件下内生真菌改善了宿主高羊茅的生长。2)与水溶性磷相比,高羊茅根有机酸和酸性磷酸酶(acid phosphatase,APase)活性在难溶性磷条件下显著增加,而根总酚含量无显著变化。在水溶性磷条件下,高羊茅EI植株根总酚含量显著高于EF植株,此时EI植株比EF植株拥有更多分蘖数和绿叶数,说明在水溶性磷条件下内生真菌对宿主地上部生长具有一定贡献。在难溶性磷条件下,虽然高羊茅EI植株根总酚含量仍然高于EF植株,但同时EI植株根有机酸含量显著低于EF植株,因此内生真菌感染只是增大了宿主植物的根冠比,而对分蘖数和绿叶数等无显著影响,说明内生真菌对宿主利用难溶性磷贡献不大。可见,内生真菌对宿主植物的生长在水溶性磷条件下更有利。     

内生真菌种类和母本基因型对内生真菌-禾草共生体叶形状和叶面积的影响

Epichloë内生真菌感染能够影响宿主植物的生长发育, 但关于内生真菌感染对宿主植物叶形状和叶面积的研究很少。该研究以羽茅(Achnatherum sibiricum)为实验材料, 采用长宽系数计算和扫描测定叶面积相结合的方法探究内生真菌种类和羽茅母本基因型对羽茅-内生真菌共生体叶形状和叶面积的影响。结果表明: 内生真菌感染与否、内生真菌种类和宿主母本基因型对反映叶形状的叶校正系数、叶片长度、宽度和长宽比均无显著影响, 经计算与验证, 确定了羽茅叶片的校正系数为0.594 9。采用该校正系数及叶长宽计算的叶面积与实测叶面积无显著差异, 且二者均未受到内生真菌感染与否、内生真菌种类或宿主植物母本基因型的显著影响。     

渗透胁迫下内生真菌感染对黑麦草幼苗生长的影响

以含有内生真菌的黑麦草(Lolium perenne L．)种子为材料,采用4C冰箱内和20℃光照培养箱内保存18个月的方式分别构建内生真菌感染(EI)和内生真菌非感染(EF)的黑麦草种群,通过比较EI和EF种群在正常条件下(对照)和渗透胁迫条件下种子发芽、幼苗生长等方面的差异,探讨内生真菌对其宿主植物的直接和间接影响。结果表明：在对照和胁迫条件下,E1种子的发芽势均明显高于EF种子,而只在重度渗透胁迫下,E1种子的发芽率才显著高于EF种子。对于黑麦草幼苗而言,渗透胁迫下内生真菌对宿主植物的地上部分和地下部分均有增益作用,最终表现为E1种群的总生物量显著高于EF种群,其中对地上部分的促进作用表现为内生真菌的存在不仅提高了宿主叶片的延伸速率、使EI叶片比EF叶片更长、叶面积更大,而且在重度胁迫下,E1种群的分蘖数也显著高于EF种群;对地下部分的促进作用表现为EI种群的根系总长度和根干重均高于EF种群。     

不同传播方式的内生真菌感染对羽茅的生理生态影响

通过田间试验, 研究了垂直传播的Neotyphodium属内生真菌和水平传播的Epichloë属内生真菌对羽茅(Achnatherum sibiricum)生理生态的影响。结果表明: Neotyphodium属内生真菌感染对羽茅的株高和叶长有显著的正效应, 而Epichloë属内生真菌感染对羽茅株高和叶长的增益作用较少。感染Neotyphodium属内生真菌的羽茅最大净光合速率显著高于感染Epichloë属内生真菌的羽茅, Neotyphodium属内生真菌显著提升了羽茅的气孔限制值和水分利用效率, 而实验测定的第一年结果显示: Epichloë属内生真菌对羽茅的各个光合指标具有明显的负效应(胞间CO₂浓度除外)。Neotyphodium属内生真菌感染使得宿主植物积累的可溶性糖含量显著高于感染Epichloë属内生真菌的羽茅和不染菌的羽茅植株。感染Epichloë属内生真菌的羽茅中氮含量显著高于感染Neotyphodium属内生真菌的羽茅。总之, 内生真菌的传播方式是影响羽茅生理生态学特性的一个因素, 并且垂直传播的内生真菌对羽茅生长和生理特性的益处高于水平传播的内生真菌。     

干旱胁迫下内生真菌感染对黑麦草光合色素和光合产物的影响

以含有内生真菌的黑麦草 (L olium perenne L.)种子为材料 ,采用加热处理方式构建内生真菌非感染的黑麦草种群 ,通过比较内生真菌感染 (EI)和非感染 (EF)植株在正常条件下和干旱胁迫条件下叶片相对水分含量、叶绿素、可溶性糖和淀粉含量等指标的差异 ,探讨黑麦草 EI和 EF种群对干旱胁迫的适应性差异。结果表明 :在中度胁迫后期 ,EI植株叶片的 RWC显著高于 EF植株 ,即 EI植株的保水能力更强。轻度水分胁迫下 ,内生真菌感染可使其宿主植物的可溶性糖含量增加 ,以增强宿主的渗透调节能力 ,随着干旱胁迫强度的加大 ,内生真菌的这一增益效应不再起作用 ,此时 ,宿主植物将更多的光合产物——淀粉积累于体内 ,以度过不良环境。第 2年春天 EI和 EF种群的恢复生长情况进一步表明 ,经过中度干旱胁迫后 ,EI种群的恢复更为迅速。生物量的大小是植物种群净光合作用能力的直接体现 ,研究中在中度干旱胁迫条件下 ,黑麦草 EI种群的生物量显著高于EF种群 ,但从光合色素的变化来看 ,相同水分状况下 EI和 EF植株的 Chla、Chlb以及 Car的变化趋势比较接近 ,这说明内生真菌感染并未缓解干旱胁迫对光合色素的破坏 ,内生真菌可能通过其它途径来改善宿主植物的光合能力     

庞泉沟自然保护区常见禾本科植物内生真菌分布及其影响因素

以庞泉沟自然保护区4种不同群落类型林地和灌丛下的优势禾草为研究对象,调查不同禾草内生真菌感染率,研究内生真菌感染的禾草种群与原生生境的土壤理化性质、土壤酶活性及土壤微生物群落结构的相互关系。结果发现,影响紫羊茅、中亚薹草和野燕麦染菌率的主要因子是土壤水分含量和碳氮比,且土壤C、N、S以及土壤水分含量与紫羊茅的内生真菌染菌率成正相关,土壤蔗糖酶、脲酶和酸性磷酸酶与紫羊茅的感染率显著正相关,但中亚薹草与紫羊茅的结果相反。丝盖伞科是华北落叶松林和油松林土壤微生物群落的优势真菌,但不同染菌植被群落类型下的优势细菌微生物群落组成不同。总体看来,4种森林群落类型下禾草内生真菌染菌率与土壤理化性质相关性不同,且受到的生态因子影响大小也有差异,内生真菌感染导致各植被群落类型的土壤微生物群落结构具有差异,其优势土壤真菌和土壤细菌组成不同,这有助于进一步认识原生生境下不同感染率的禾草种群对土壤微生物群落结构的影响及其在生态系统中的各异生态学功能。     

Effects of the endophyte Epichloë coenophiala on the root microbial community and growth performance of tall fescue in different saline-alkali soils

Soil salinization is detrimental to plant growth and yield in agroecosystems worldwide. Epichloë endophytes, a class of clavicipitaceous fungi, enhance the resistance of host plants to saline-alkali stress. This study explored the effects of the systemic fungal endophyte Epichloë coenophiala on the root microbial community and growth performance of tall fescue (Lolium arundinaceum) growing under different saline-alkali stress conditions. Structural equation modeling (SEM) was conducted to analyze the direct and indirect effects (mediated by root microbial community diversity and soil properties) of the endophyte on the growth of tall fescue under saline-alkali stress. The endophyte-infected plants produced higher shoot and root biomass compared to endophyte-free plants under saline-alkali stress (200 and 400 mM). Endophyte infection increased the fungal community diversity and altered its composition in the roots, decreasing the relative abundance of Ascomycota and increasing that of Glomeromycota. Furthermore, endophyte infection decreased the bacterial community diversity and the relative abundance of dominant Proteobacteria. SEM showed that endophyte infection increased the shoot and root biomass under saline-alkali stress (200 and 400 mM) by increasing the arbuscular mycorrhizal fungal diversity in the roots, and soil total nitrogen and phosphorus concentrations. Therefore, it is important to examine aboveground microbes as factors influencing plant growth in saline-alkali stress by affecting belowground microbes and soil chemical properties.     

Effects of endophytic fungi on some drought tolerance mechanisms of tall fescue in a hydroponics culture

Neotyphodium, a seed-transmissible nonpathogenic fungal endophyte (symbiont) is considered beneficial because endophyte-infected grasses are more drought-tolerant, produce more dry matter, utilize soil nitrogen more efficiently, and deter insects. In this study, the effects of endophytes on physiological mechanisms of drought tolerance in tall fescue (Festuca arundinacea Schreb.) were studied in a greenhouse. Two clonally propagated genotypes of tall fescue (F. arundinacea Schreb.), naturally containing endophyte (EI), and their endophyte-free ramets (EF) were tested at three water stress treatments exerted by PEG 6000 in a hydroponics system. Relative water content (RWC), cell membrane stability (CMS), proline and chlorophyll contents in plant leaves were measured during water stress treatments. After harvest, K⁺, Ca²⁺, and Mg²⁺ contents were measured in plant roots and shoots. After 20 days under stress conditions, plants were transferred to basal hydroponics medium, and their survival after stress relief was evaluated. The results showed that endophyte considerably contributes to host grass water stress tolerance. Both genotypes of EI and EF plants did not differ in RWC, but, regardless of the infection status, genotype 75 had the higher RWC than genotype 83. EI clones of both genotypes maintained slightly higher chlorophyll content and membrane stability than EF clones, although these differences were not significant. The EI plants of genotype 83 concentrated significantly more proline than EF plants, but in the genotype 75, differences between EI and EF clones were not significant. Plant mineral absorption was also influenced by the endophyte presence. EI clones had the higher concentrations of K⁺ in the shoots of both genotypes. The Mg²⁺ and Ca²⁺ contents in EF plants of both genotypes were higher than EI plants in the roots, but in the shoots there were no differences between EI and EF clones. EI clones survived longer after stress removal. These results strongly suggest that Neotyphodium endophytes exert their effects on tall fescue drought tolerance through alteration of various physiological mechanisms involved. Published in Russian in Fiziologiya Rastenii, 2009, Vol. 56, No. 4, pp. 563–570. This test was submitted by the authors in English.     

Neotyphodium Coenophialum-Infected Tall Fescue and Its Potential Application in the Phytoremediation of Saline Soils

The growth response of endophyte-infected (EI) and endophyte-free (EF) tall fescue to salt stress was investigated under two growing systems (hydroponic and soil in pots). The hydroponic experiment showed that endophyte infection significantly increased tiller and leaf number, which led to an increase in the total biomass of the host grass. Endophyte infection enhanced Na accumulation in the host grass and improved Na transport from the roots to the shoots. With a 15 g l^?1 NaCl treatment, the phytoextraction efficiency of EI tall fescue was 2.34-fold higher than EF plants. When the plants were grown in saline soils, endophyte infection also significantly increased tiller number, shoot height and the total biomass of the host grass. Although EI tall fescue cannot accumulate Na to a level high enough for it to be termed a halophyte, the increased biomass production and stress tolerance suggested that endophyte / plant associations had the potential to be a model for endophyte-assisted phytoextraction in saline soils.     

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。     

Genotypic and chemotypic diversity of Neotyphodium endophytes in tall fescue from Greece

Epichloid endophytes provide protection from a variety of biotic and abiotic stresses for cool-season grasses, including tall fescue. A collection of 85 tall fescue lines from 15 locations in Greece, including both Continental and Mediterranean germplasm, was screened for the presence of native endophytes. A total of 37 endophyte-infected lines from 10 locations were identified, and the endophytes were classified into five distinct groups (G1 to G5) based on physical characteristics such as colony morphology, growth rate, and conidial morphology. These classifications were supported by phylogenetic analyses of housekeeping genes tefA and tubB, and the endophytes were further categorized as Neotyphodium coenophialum isolates (G1, G4, and G5) or Neotyphodium sp. FaTG-2 (Festuca arundinacea taxonomic group 2 isolates (G2 and G3). Analyses of the tall fescue matK chloroplast genes indicated a population-wide, host-specific association between N. coenophialum and Continental tall fescue and between FaTG-2 and Mediterranean tall fescue that was also reflected by differences in colonization of host tillers by the native endophytes. Genotypic analyses of alkaloid gene loci combined with chemotypic (chemical phenotype) profiles provided insight into the genetic basis of chemotype diversity. Variation in alkaloid gene content, specifically the presence and absence of genes, and copy number of gene clusters explained the alkaloid diversity observed in the endophyte-infected tall fescue, with one exception. The results from this study provide insight into endophyte germplasm diversity present in living tall fescue populations.     

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。     

醉马草内生真菌共生体研究进展

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。     

Endophytes inconsistently affect plant communities across Schedonorus arundinaceus hosts

Fungal endophytes in cool-season grasses may affect communities at multiple trophic levels. However, it is unclear whether community-scale endophyte effects arise due to the endophyte itself or as a result of unique, endophyte–host interactions. We used a long-term field experiment to test whether common-toxic (CT) and non-ergot alkaloid-producing (novel) endophytes in Schedonorus arundinaceus (tall fescue) forage cultivars consistently affect communities across tall fescue hosts. Tilled plots (2 × 2 m; Guelph, ON) were seeded with Georgia 5 and Jesup cultivars containing either the CT or AR542 (novel) endophyte and allowed to be re-colonized by plant species from the local propagule pool. Non-seeded control plots were included to assess effects of seeding the non-native grass. We assessed plant, invertebrate, soil moisture, and soil nutrient responses to the endophyte–cultivar treatments after four growing seasons. Seeding tall fescue affected plant species abundances, but not richness, and did not consistently alter soil moisture and nutrient pools. Endophyte identity in the tall fescue cultivars affected the communities, but effects were not consistent between cultivars. Within Georgia 5, the AR542 endophyte reduced tall fescue abundance and altered the invertebrate community relative to CT plots. Within Jesup, the AR542 endophyte reduced species evenness and decreased soil moisture during dry periods relative to CT plots. Endophyte effects were not consistent between cultivars, and it is probable that the community-scale effects of endophyte infection in tall fescue cultivars arise due to unique interactions between cultivar and endophyte.     

Endophytic fungus improves growth and metal uptake of Lolium arundinaceum Darbyshire ex. Schreb

The effect of endophyte infection on plant growth, cadmium (Cd) uptake, and Cd translocation was investigated using tall fescue (Lolium arundinaceum) grown in greenhouses in contaminated solution. Endophyte infection significantly increased tiller number and biomass of the host grass under both control and Cd-stress conditions. Endophyte infection not only enhanced Cd accumulation in tall fescue, but also improved Cd transport from the root to the shoot. Under 20 mg L(-1) Cd stress, the phytoextraction efficiency of endophyte-infected (EI) tall fescue was 2.41-fold higher than endophyte-free plants. Although the total Cd accumulation in EI tall fescue was insufficient for practical phytoextraction applications, the observed high biomass production and tolerance of stress from abiotic factors including heavy metals, gives endophyte/plant associations the potential to be a model for endophyte-assisted phytoremediation of metal-polluted soils.