黑麦草-内生真菌共生体对磷缺乏的生理生态反应

选取感染和未感染的黑麦草为材料,在田间盆栽条件下研究内生真菌感染对宿主植物抵抗磷胁迫方面的贡献。结果表明,土壤中缺磷或内生真菌感染对黑麦草地上部生长的影响不显著,但内生真菌感染对植株地下部生长和生理指标有明显影响。缺磷条件下,内生真菌感染有助于黑麦草地下部分的生长,表现在根系总长度更长,生物量更大;同时根中酚类物质和有机酸的含量也显著高于未感染植株,但因酚类物质和有机酸总量增加的同时并未伴随着二者浓度的增加,由此推测,内生真菌在改变宿主黑麦草根系代谢活动方面的贡献有限。此外,内生真菌感染显著提高了宿主植物的磷利用效率,这可能和缺磷条件下内生真菌感染植株具有更高的酸性磷酸酶活性有关。     

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pH砖红壤浸提液和内生真菌对黑麦草幼苗生长生理的影响

以带内生真菌(E+)和不带内生真菌(E-)的多年生黑麦草(Lolium perenne)种子为材料,采用室内种子萌发实验,分析多年生黑麦草在不同pH(4.47~6.12)砖红壤浸提液中种子萌发、幼苗形态指标及生理指标的变化,探讨内生真菌对酸性环境中多年生黑麦草幼苗生长和生理特性的影响。结果表明:(1)在相同带菌情况下(内生真菌侵染E+或未浸染E-),随着砖红壤浸提液pH值的降低(浸提液酸性的增强),E+和E-种子发芽率、发芽势、发芽指数、活力指数、胚芽长、胚根长、胚芽鲜重和胚芽干重均表现为逐渐下降的趋势,且5个砖红壤浸提液处理均显著低于相应对照(CK,pH 6.12);幼苗脯氨酸含量、超氧化物歧化酶活性、过氧化物酶活性和丙二醛含量呈上升趋势,且5个砖红壤浸提液处理均显著高于相应CK。(2)除CK外,在相同pH砖红壤浸提液处理下,内生真菌侵染显著提高了多年黑麦草的发芽率、胚芽长、鲜重和干重等形态指标及脯氨酸含量和超氧化物歧化酶活性,却显著降低了过氧化物酶活性和丙二醛含量;在砖红壤浸提液pH为4.47~6.12时,E+萌发种子胚根长与E-之间无显著差异。研究发现,不同pH砖红壤浸提液对E+和E-种子萌发、生长和生理有一定的影响,内生真菌的侵染可促进宿主植物的生长,同时对砖红壤浸提液pH值具有较宽的适应范围,可将多年生黑麦草内生真菌共生体应用于酸性土壤改良。     

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。     

不同氮素营养条件下内生真菌感染对黑麦草光合和蒸腾速率及生物量的影响

以黑麦草(Lolium perenne L.)为实验材料,研究在不同氮素营养条件下内生真菌(Neotyphodium lolii)感染对其光合特性及生物量的影响。结果表明：1)整个实验中,内生真菌感染对黑麦草净光合速率、蒸腾速率以及水分利用效率均无显著影响,但在中氮和高氮水平时,感染种群的净同化累积值有超过非感染种群的趋势;2)在本实验的第一处理期和第二处理期,内生真菌感染对黑麦草绿色部分的干物质产量无显著影响,但在第三处理期感染种群的绿色部分的干物质产量显著高于非感染种群。     

不同氮素营养条件下内生真菌感染对黑麦草光合和蒸腾速率及生物量的影响

以黑麦草(Lolium perenne L)为实验材料,研究在不同氮素营养条件下内生真菌(Neotyphodium lolii)感染对其光合特性及生物量的影响.结果表明:1)整个实验中,内生真菌感染对黑麦草净光合速率、蒸腾速率以及水分利用效率均无显著影响,但在中氮和高氮水平时,感染种群的净同化累积值有超过非感染种群的趋势;2)在本实验的第一处理期和第二处理期,内生真菌感染对黑麦草绿色部分的干物质产量无显著影响,但在第三处理期感染种群的绿色部分的干物质产量显著高于非感染种群.     

不同磷水平土壤接种丛枝菌根真菌对植物生长和养分吸收的影响

通过盆栽试验研究了不同磷水平土壤接种丛枝菌根真菌Glomus mosseae对三叶草(Clover)和黑麦草(Ryegrass)生长、P浓度及N ∶ P比的影响.试验结果表明,Glomus mosseae显著提高了三叶草中P浓度和生物量,降低了植株N ∶ P比,对黑麦草植株P含量、生物量及N ∶ P比没有显著影响;在不同磷水平土壤中,接种处理对单种时三叶草中P含量的影响程度不同,以中磷和高磷水平下的影响较显著,这表明Glomus mosseae对三叶草的菌根效应在不同磷水平下并不相同;另外,两种植物混合种植情况下,黑麦草的竞争能力高于三叶草,使得其P浓度和生物量都显著高于三叶草,而接种Glomus mosseae增加了混种中三叶草的生物量和P浓度,降低了黑麦草的生物量,表明Glomus mosseae能提高混种中三叶草的竞争能力,从而改变群落中物种的竞争关系.此外,在一定磷水平下,菌根真菌的存在降低了三叶草中N ∶ P比, 这会改变三叶草的受限制方式,由受P元素限制变为受N元素限制.     

施肥对苗期紫茎泽兰和黑麦草相对竞争力的影响

为种植黑麦草替代控制紫茎泽兰提供依据,运用取代实验法,研究了施肥(氮、磷)对苗期紫茎泽兰和黑麦草的相对竞争力以及生长表现的影响。结果表明:增施氮肥和磷肥均能够提高紫茎泽兰的相对竞争力,而仅磷肥对黑麦草的竞争力略有促进作用;但在各种施肥水平下,黑麦草的竞争力仍然明显强于紫茎泽兰。增施氮肥可以显著提高紫茎泽兰的株高、分枝数和干质量,而磷肥仅在较高时才显著提高其干质量;增施氮肥虽然可以提高黑麦草的分蘖数量,但对其干质量没有影响,而增施磷肥(2次)可以显著提高其干质量;在竞争中紫茎泽兰植株生长的能量分配(用根茎比表示)对磷肥不敏感。建议适当增施磷肥、不施或少施氮肥,以提高黑麦草的替代控制效果。     

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。     

内生真菌感染对干旱胁迫下黑麦草生长的影响

 内生真菌是生活在健康植物的茎叶内,形成不明显感染的一类真菌。以黑麦草（Lolium perenne L.）为实验材料,研究在不同强度的干旱胁迫下内生真菌（Neotyphodium lolii）侵染对其叶片延伸生长、分蘖数和生物量的影响。结果表明,与非感染种群相比,内生真菌感染对黑麦草叶片延伸速率无明显促进作用;内生真菌感染种群具有明显较多的分蘖数;在重度胁迫并经过恢复期后,内生真菌感染种群具有较高的根冠比。因而内生真菌可能通过提高植物的分蘖能力和促进有机物向根系的分配来促进宿主植物的营养生长并提高其抗旱性     

Endophytic fungi (Neotyphodium coenophialum) affect the growth and mineral uptake, transport and efficiency ratios in tall fescue (Festuca arundinacea)

Neotyphodium coenophialuminteracts mutualistically with its host grasses. Tall fescue (Festuca arundinacea Schreb.) plants infected by the fungal endophyte,Neotyphodium coenophialum(Morgan-Jones and Gams) Glenn, Bacon and Hanlin, often perform better than non-infected plants, especially in limited resource environments. However, there is a scarcity of information about endophyte-grass ecotypes interaction in Andisols of temperate regions. Clones of three tall fescue ecotypes (Fukaura, Koiwai and Showa) either infected with N. coenophialum (E+) or noninfected (E–) were grown in Andisols (Black Andisol: naturally low content of phosphorus, high in other nutrients; Red Andisol: naturally high content of phosphorus, low in other nutrients) for 133 days in a controlled environment. Cumulative shoot dry weight, daily regrowth rates (tiller number, plant height and shoot dry matter) after clippings and nutrient uptake, transport and efficiency ratios were measured. In Black Andisol, E+ plants had significantly higher cumulative shoot dry weight as well as daily regrowth rates than E– plants, while in Red Andisol the reverse was true. Among the ecotypes studied, Showa had the highest shoot growth. Significantly higher phosphorus (P), potassium (K), calcium (Ca) and magnesium (Mg) uptake as well as transport were identified in E+ vs. E– plants grown in Black Andisol. With few exceptions, values for nutrient efficiency ratios were not significantly different between E+ and E– plants grown in both soils. Significant three-way interaction (endophyte × ecotype × soil) for cumulative shoot dry weight and regrowth rate revealed that the ecotype specific regrowth responses to endophyte infection were depended on soil nutrient conditions. Vegetative growth and nutrient acquisition in tall fescue varied with ecotype and were modified by abiotic (soil fertility status) as well as biotic (endophyte infection) factors.     

Leaf endophyte Neotyphodium coenophialum modifies mineral uptake in tall fescue

Neotyphodium coenophialum (Morgan-Jones and Gams) Glenn, Bacon and Hanlin, a fungal endophyte found primarily in shoots of tall fescue (Festuca arundinacea Shreb.), can modify rhizosphere activity in response to phosphorus (P) deficiency. In a controlled environment experiment, two cloned tall fescue genotypes (DN2 and DN4) free (E-) and infected (E+) with their naturally occurring endophyte strains were grown in nutrient solutions at low P (3.1 ppm) or high P (31 ppm) concentrations for 21 d. Endophyte infection increased root dry matter (DM) of DN4 by 21% but did not affect root DM of DN2. Under P deficiency, shoot and total DM were not affected by endophyte but relative growth rate was greater in E+ than E- plants. In high P nutrient solution, E+ plants produced 13% less (DN2) or 29% more (DN4) shoot DM than E- plants. Endophyte affected mineral concentrations in roots more than in shoots. Regardless of P concentration in nutrient solution, E+ DN2 accumulated more P, Ca, Zn and Cu but less K in roots than E- plants. When grown in high P nutrient solution, concentrations of Fe and B in roots of E+ DN2 plants were reduced compared with those of E- plants. Concentrations of P, Ca and Cu in roots of DN4 were less, but K was greater in E+ than E- plants. In shoots, E+ DN2 had greater concentrations of Fe and Cu than E- DN2, regardless of P concentration in nutrient solution. Genotype DN4 responded to endophyte infection by reducing B concentration in shoots. Nutrient uptake rates were affected by endophyte infection in plants grown in low P nutrient solution. A greater uptake rate of most nutrients and their transport to shoots was observed in DN2, but responses of DN4 were not consistent. Results suggest that endophyte may elicit different modes of tall fescue adaptation to P deficiency. This revised version was published online in June 2006 with corrections to the Cover Date.     

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。     

Effects of simultaneous infections of endophytic fungi and arbuscular mycorrhizal fungi on the growth of their shared host grass Achnatherum sibiricum under varying N and P supply

Grasses can be infected by endophytic fungi and arbuscular mycorrhizal (AM) fungi simultaneously. Here, we investigated the interactions of a native grass, Achnatherum sibiricum, with both endophytic and AM fungi (Glomus mosseae, GM and Glomus etunicatum, GE) at different nitrogen (N) and phosphorus (P) levels. The results showed that endophyte infection significantly suppressed the colonization rates and spore density of GE, but had no effect on those of GM. Endophyte infection increased shoot biomass regardless of the nutrient conditions. The effects of AM fungi on host growth were dependent on mycorrhizal species. There was no significant interaction between endophytic fungi and GE on host growth; however, a significant interaction between endophytic fungi and GE occurred in total phenolic concentrations and P uptake. As for GM, a significant interaction among endophytic fungi, AM fungi and nutrient availability occurred in shoot growth. Under sufficient N and P conditions, endophyte infection alleviated the detrimental effects of GM colonization on host growth.     

Plant population and genotype effects override the effects of Epichloë endophyte species on growth and drought stress response of Achnatherum robustum plants in two natural grass populations

Aims In cool-season grasses, systemic and vertically transmitted Epichlo? infections often provide a suite of benefits including increased growth, reproduction and competitive abilities. However, these effects of Epichlo? endophytes on their hosts often depend upon host and endophyte genotype and environmental factors.Methods Achnatherum robustum (sleepygrass) harbors at least two Epichlo? species within natural populations in the Southwest USA. We tested the effects of endophyte infection and species, host population and plant genotype (by experimentally removing the endophyte), and soil moisture (a key limiting factor) on growth and drought stress response of infected A. robustum plants from two populations (Weed and Cloudcroft) in the Sacremento Mountains of New Mexico, USA).Important findings Although the two populations harbor distinct Epichlo? species each with very different chemoprofiles, neither endophyte status (infected vs. uninfected) nor endophyte species affected most growth parameters at 8 or 25 weeks of the experiment, except for leaf length. In high water treatment, infected plants from the Weed population had longer leaf length compared with uninfected plants. In contrast, the population of origin affected all growth parameters, including plant height, leaf number, length and width, tiller number and shoot and root biomass, as well as wilting time. Grasses from the Cloudcroft population generally showed greater growth than grasses from the Weed population. Endophyte infection did affect wilting time, with infection in the Weed population generally reducing time to wilting under low and high water, whereas infection in the Cloudcroft population reduced time to wilting only under high water conditions. Our results suggest that plant population and their associated plant genotypes may play a much larger role in endophyte–host grass interactions in varying environments than previously thought. Asexual Epichlo? species may be compatible with only specific host genotypes within populations such that the phenotypic effects due to population may be greater than phenotypic changes influenced by variation in the endophyte.     

Environmental conditions and host genotype direct genetic diversity of Venturia ditricha,a fungal endophyte of birch trees

We investigated whether genetic variation of a common foliar endophyte of birch trees, Venturia ditricha, is affected by environmental conditions or host genotype. Fungal samples were collected from 10 half-sibling families of mountain birch (Betula pubescens ssp. czerepanovii) grown in two environmental conditions with different daily average temperatures: a forested river valley and an adjacent open tundra (altitudinal difference 180 m). Genetic analysis of V. ditricha isolates was done using random amplified microsatellite polymerase chain reaction. We found that host genotypes, along with prevailing environmental conditions, influence the probability of infection by particular endophyte genotypes. The most susceptible host genotypes were highly infected with genetically similar endophyte genotypes, whereas the most resistant trees were poorly infected and they were infected by genetically dissimilar endophytes. Our results also showed environment-host genotype interactions, suggesting that the susceptibility of the host to a particular endophyte genotype may change in natural environments when environmental conditions are changed. It appears that a particular endophyte genotype needs to find the right host genotype for a successful infection. There are many host genotypes in natural stands; this means, from the point of view of the fungus, the environment is heterogeneous. Thus, under the influence of birch tree genotypes, genetically differentiated subgroups of the endophytic fungus may be formed in different environments.     

Effects of endophyte infection on drought stress tolerance of Lolium perenne accessions from the Mediterranean region

Perennial ryegrass (Lolium perenne) is a cool-season grass species that is often infected by the leaf-inhabiting endophyte Neotyphodium lolii. This particular endophyte is asexual and has the potential to impact host survival, growth and reproduction. The objective was to assess the potential costs or benefits of endophyte infection on drought stress tolerance of native perennial ryegrass accessions originally collected from Italy, Morocco, Tunisia, and Turkey. Sixty infected (E+) individuals from each accession were planted in a greenhouse. Half of these individuals were treated with a systemic fungicide to eliminate the endophyte (E−). For two drought periods water was withheld for 10-14 days and then allowed a one week recovery period following each. In some accessions under drought, E+ plants had more tillers, greater tiller lengths, total dry mass and green shoot mass than E− plants, suggesting a positive effect of endophyte infection on host growth. Total tiller length and the number of tillers showed significant population × treatment × infection interactions for 4 of 6 populations. This work is one of the few that documents the effects of endophyte infection for a common forage grass species from wild populations native to its distributional range. The results demonstrate that endophyte infection can help ameliorate abiotic stress such as drought and there may be a selective advantage for grasses from certain Mediterranean regions.     

Interaction between fungal endophytes and environmental stressors influences plant resistance to insects

Neotyphodium coenophialum, an endophytic fungus that infects shoots of tall fescue (Festuca arundinacea), may protect its host from herbivory through production of alkaloids. Yet, the fungus can also modify plant resource allocation, regrowth dynamics, and drought tolerance, and these changes may also influence herbivores. We tested if N. coenophialum infection interacted with stress (drought or simulated herbivory) to modify plant resistance to insects. We assigned greenhouse plants to one of four treatments: 1) clipping at 3 cm above the soil surface, 2) drought stress during insect bioassays, 3) drought stress prior to insect bioassays, or 4) daily watering. Treatments were crossed with presence or absence of endophyte to give eight treatment combinations, and we assessed the performance of bird cherry‐oat aphid (Rhopalosiphum padi) and fall armyworm (Spodoptera frugiperda) feeding on plants in two separate experiments from each of the eight treatments. Aphids were placed into clip bags on leaf blades and allowed to reproduce parthenogenetically. Plant tissue was fed to third instar fall armyworm caterpillars until they molted into the fifth instar. Developmental time was recorded and larval growth was obtained gravimetrically. We also assessed total protein nitrogen (N) and loline alkaloids in plants.
Total protein N was unaffected by endophyte infection. In contrast, stress influenced total protein N, but its effect varied with endophyte infection. Uninfected plants that were clipped had higher total protein N; this trend was absent in infected plants. Plants in drought stress had lower N, but only if they were infected. Lolines were nearly absent from uninfected plants. In infected plants they tended to be higher in clipped plants. The effect of endophyte infection differed between the two insects: aphid reproduction was reduced by the endophyte, but endophyte infection enhanced caterpillar performance. Both insects were affected by interactions between the endophyte and stress. Aphids were negatively affected by drought stress, but only when feeding on uninfected plants, while caterpillars showed the opposite response, displaying lower performance on drought stressed plants only if they were infected. Aphids reproduced faster on regrowth tissue (following damage by clipping) of uninfected plants, but endophyte infection cancelled this effect. In contrast, performance of caterpillars was not influenced by an interaction between damage and infection. We conclude that N. coenophialum does not provide universal resistance to insects. Endophyte‐mediated resistance varies with insect species and will be a complex function of environmental stress, including drought and prior damage.     

Effects of the Neotyphodium endophyte fungus on dormancy and germination rate of Lolium multiflorum seeds

Abstract Neotyphodium frequently occurs as an endophyte in grasses. Evidence shows enhanced fitness of endophyte infected grasses relative to non‐infected ones. Some studies of seed germination show endophyte enhancement of plant fitness in various environments, but inconsistent results indicate that further studies are needed. So far, experiments have failed to separate the confounded effects of population origin and seed management. For this reason, we evaluated the effects of endophyte infection on seed dormancy and germination in Lolium multiflorum using an experimental design controlling these factors. Depending on the year of seed production, endophyte infection modified seed response to light quality, affecting predominantly seed dormancy levels. Nevertheless, the endophyte did not affect base temperature or thermal time of germination. We concluded that endophytes were not a strong influence on germination behaviour. We speculate from our results that the presence of the endophyte changes germination by an indirect effect, in extending growth of the maternal plant during seed development and ripening. The direct effect of hyphae in the seed on seed behaviour was disregarded, because the difference between infected and non‐infected seed varied within the year of seed production. Future experiments should focus on effects of the endophyte on the canopy of parent plants during seed production and ripening, and, hence, on subsequent dormancy and germination of the seeds.