Plant sesquiterpenes induce hyphal branching in arbuscular mycorrhizal fungi

Arbuscular mycorrhizal (AM) fungi form mutualistic, symbiotic associations with the roots of more than 80% of land plants. The fungi are incapable of completing their life cycle in the absence of a host root. Their spores can germinate and grow in the absence of a host, but their hyphal growth is very limited. Little is known about the molecular mechanisms that govern signalling and recognition between AM fungi and their host plants. In one of the first stages of host recognition, the hyphae of AM fungi show extensive branching in the vicinity of host roots before formation of the appressorium, the structure used to penetrate the plant root. Host roots are known to release signalling molecules that trigger hyphal branching, but these branching factors have not been isolated. Here we have isolated a branching factor from the root exudates of Lotus japonicus and used spectroscopic analysis and chemical synthesis to identify it as a strigolactone, 5-deoxy-strigol. Strigolactones are a group of sesquiterpene lactones, previously isolated as seed-germination stimulants for the parasitic weeds Striga and Orobanche. The natural strigolactones 5-deoxy-strigol, sorgolactone and strigol, and a synthetic analogue, GR24, induced extensive hyphal branching in germinating spores of the AM fungus Gigaspora margarita at very low concentrations.     

菌根共生体中植物信号物质的产生及其作用机制

菌根是丛枝菌根真菌与宿主植物形成的互利共生体,在共生的不同阶段,植物产生一系列的信号物质如独脚金内酯、溶血磷脂酰胆碱、茉莉酸和水杨酸等.这些信号物质在植物与丛枝菌根真菌间相互识别、菌根形成、营养物质交换,以及植物防御反应诱导等方面发挥重要作用.文中探讨植物信号物质在丛枝菌根共生系统形成前后和病原体侵害的3个阶段可能的生理效应和作用机制,旨在为研究丛枝菌根真菌与植物信号物质的相互作用及其在农业生产和环境保护方面的应用提供依据.     

Arbuscular mycorrhizal formation of crucifer leaf mustard induced by flavonoids apigenin and daidzein

Flavonoids from legume root secretion may probably act as signal molecules for expression of Rhizobial “nod” nodulation genes and AM fungal symbiotic gene. Leaf mustard is a non-mycorrhizal plant; it does not contain flavonoids and other signal molecules. AM fungi could not infect the roots of leaf mustard and form a symbiont in nature,when it was treated with flavonoids (apigenin or daidzein).The results of trypan blue staining showed that two kinds of AM fungi (G intraradices and G mosseae) successfully infected the roots of non-mycorrhizal plant leaf mustard. AM fungi grew towards and colonized the roots of leaf mustard,producing young spores and completing the course of life.AM fungi are the only one kind of fungi with ALP activity.The result of ALP staining has also proved that AM fungi infected successfully the roots of leaf mustard. AM fungi (G intraradices and G mosseae) that existed in the roots of non-mycorrhizal plant leaf mustard were probed by nested PCR and special molecular probes. The above-mentioned proof chains have fully proved that flavonoids induced AM fungi (G intraradices and G mosseae) to infect non-mycorrhizal plant and establish symbiotic relationship.     

A plant receptor-like kinase required for both bacterial and fungal symbiosis

Most higher plant species can enter a root symbiosis with arbuscular mycorrhizal fungi, in which plant carbon is traded for fungal phosphate. This is an ancient symbiosis, which has been detected in fossils of early land plants. In contrast, the nitrogen-fixing root nodule symbioses of plants with bacteria evolved more recently, and are phylogenetically restricted to the rosid I clade of plants. Both symbioses rely on partially overlapping genetic programmes. We have identified the molecular basis for this convergence by cloning orthologous SYMRK ('symbiosis receptor-like kinase') genes from Lotus and pea, which are required for both fungal and bacterial recognition. SYMRK is predicted to have a signal peptide, an extracellular domain comprising leucine-rich repeats, a transmembrane and an intracellular protein kinase domain. Lotus SYMRK is required for a symbiotic signal transduction pathway leading from the perception of microbial signal molecules to rapid symbiosis-related gene activation. The perception of symbiotic fungi and bacteria is mediated by at least one common signalling component, which could have been recruited during the evolution of root nodule symbioses from the already existing arbuscular mycorrhiza symbiosis.     

几种野生花卉丛枝菌根发育状况的研究

丛枝菌根是自然界中,植物与真菌形成的分布最广泛的一类互惠共生体。作者对山东省的6种重要野生花卉小黄花菜(Hemerocallis minorMill.)、萱草(Hemerocallis fulvaL.)、琥珀千里光(Senecio ambraceusTurcz.)、矮鸢尾(Iris chamaeiris)、甘野菊(Dendranthema lavandulaefolium)、锦鸡儿(Catagana sinicaRehd.)的AM真菌的侵染状况、典型结构的发育状况以及其根际土壤中AM真菌的孢子密度和根际AM真菌的相对多度和频度进行了研究。结果表明,所调查的6种野生花卉都被AM真菌侵染,且可观察到典型结构泡囊,其根际土壤中AM真菌孢子密度也相当丰富。在本次调查中共分离到了5个属的AM真菌,其中无梗囊霉属和球囊霉属真菌是野生花卉根际AM真菌的优势类群。     

丛枝菌根真菌在农业中的应用：研究进展与挑战

 丛枝菌根（arbuscular mycorrhiza，AM）真菌能够和大多数农作物形成互利共生体系，有效改善作物矿质营养，减轻环境胁迫对作物生长的不利影响，提高作物产量和品质。由于AM真菌的功能多面性，其在农业生产中具有潜在重要作用和广阔应用前景。结合AM真菌农业应用技术途径和适用情景，讨论了作物种植制度和管理措施对AM真菌功能的影响及可能的调控途径，分析了菌根技术发展趋势及面临的机遇和挑战，提出加强AM真菌功能多面性及其应用途径优化研究，将有助于促进AM真菌菌剂相关产业的发展以及AM真菌在农业生产中的广泛应用。     

用分子探针探测两种AM真菌在同一宿主植物根内的侵染

两种AM菌根真菌(G．intraradices和G．mosseae)混合接种于同一宿主植物紫云英中，通过Trypan blue染色检测了AM真菌在紫云英根中的存在，并通过nested PCR和特异性的分子探针，探测了Gintraradices和G．mosseae对紫云英同一根段的侵染。     

Shoot control of root development and nodulation is mediated by a receptor-like kinase

In legumes, root nodule organogenesis is activated in response to morphogenic lipochitin oligosaccharides that are synthesized by bacteria, commonly known as rhizobia. Successful symbiotic interaction results in the formation of highly specialized organs called root nodules, which provide a unique environment for symbiotic nitrogen fixation. In wild-type plants the number of nodules is regulated by a signalling mechanism integrating environmental and developmental cues to arrest most rhizobial infections within the susceptible zone of the root. Furthermore, a feedback mechanism controls the temporal and spatial susceptibility to infection of the root system. This mechanism is referred to as autoregulation of nodulation, as earlier nodulation events inhibit nodulation of younger root tissues. Lotus japonicus plants homozygous for a mutation in the hypernodulation aberrant root (har1) locus escape this regulation and form an excessive number of nodules. Here we report the molecular cloning and expression analysis of the HAR1 gene and the pea orthologue, Pisum sativum, SYM29. HAR1 encodes a putative serine/threonine receptor kinase, which is required for shoot-controlled regulation of root growth, nodule number, and for nitrate sensitivity of symbiotic development.     

Nitrogen fixation. Endocrine disrupters and flavonoid signalling

Nitrogen fixation is a symbiotic process initiated by chemical signals from legumes that are recognized by soil bacteria. Here we show that some endocrine-disrupting chemicals (EDCs), so called because of their effect on hormone-signalling pathways in animal cells, also interfere with the symbiotic signalling that leads to nitrogen fixation. Our results raise the possibility that these phytochemically activated pathways may have features in common with hormonal signalling in vertebrates, thereby extending the biological and ecological impact of EDCs.     

地道药材凤丹根际AM真菌与丹皮酚含量的相关性分析

凤丹(Paeonia ostii)含有重要药用价值的次生代谢产物丹皮酚.丛枝菌根真菌(AM真菌)与凤丹存在密切的共生关系,菌根真菌能够直接或间接影响植物次级代谢过程.采用HPLC法测定凤丹中丹皮酚的含量、碱解离-酸性品红染色法测定侵染率、曲利苯兰染色法测定菌丝量、湿筛沉淀法测定孢子密度.结果发现,凤丹根际丛枝菌根侵染率、孢子密度和菌丝量与丹皮酚含量呈显著的正相关.本文通过AM真菌的孢子密度、菌丝量、侵染率与凤丹丹皮酚含量的相关性分析,表明AM真菌是凤丹中丹皮酚含量的促进因素,凤丹体内AM真菌侵染越强,根际土壤中AM孢子产生越多,土壤菌丝量越高,则丹皮酚含量积累越高.研究AM真菌对丹皮酚次级代谢物生成的影响,对进一步研究AM真菌对丹皮酚在药用价值方面提供重要依据.     

Plastid proteins crucial for symbiotic fungal and bacterial entry into plant roots

The roots of most higher plants form arbuscular mycorrhiza, an ancient, phosphate-acquiring symbiosis with fungi, whereas only four related plant orders are able to engage in the evolutionary younger nitrogen-fixing root-nodule symbiosis with bacteria. Plant symbioses with bacteria and fungi require a set of common signal transduction components that redirect root cell development. Here we present two highly homologous genes from Lotus japonicus, CASTOR and POLLUX, that are indispensable for microbial admission into plant cells and act upstream of intracellular calcium spiking, one of the earliest plant responses to symbiotic stimulation. Surprisingly, both twin proteins are localized in the plastids of root cells, indicating a previously unrecognized role of this ancient endosymbiont in controlling intracellular symbioses that evolved more recently.     

云南寻甸石漠化土壤易氧化碳对丛枝菌根真菌共生的响应

【目的】运用菌根技术改良石漠化土壤性状,已成为石漠化地区植被与土壤恢复的重要生物学途径。揭示丛枝菌根(AM)真菌与植物共生驱动下石漠化土壤碳库及养分状况的变化,探明石漠化土壤易氧化碳(ROC)对土壤碳库及养分变化的响应过程及机制,为石漠化土壤的微生物修复及提高石漠化治理效率提供参考。【方法】采集云南昆明寻甸石漠化土壤,以尼泊尔桤木(Alnus nepalensis)为寄主植物,分别接种摩西斗管囊霉(Funneliformis mosseae, FM)、幼套近明球囊霉(Claroideoglomus etunicatum, CE)、根内根孢囊霉(Rhizophagus intraradices, RI),并设置对照处理(无寄主植物及无AM接种),采用高锰酸钾氧化法测定不同试验处理下土壤ROC含量,探究土壤ROC与碳库组成、养分状况及植物生长之间的相互关系。【结果】不同AM真菌均具有显著的侵染与促生效应,其中RI的侵染率与菌丝侵染密度最大,相较于对照分别提高155%和100%,并显著促进桤木树高(60%)与基径(46%)生长;不同AM真菌均显著提高ROC含量,3菌种对ROC含量的提升率大小顺序为:RI(122%)> CE(78%)> FM(61%)。ROC在土壤总有机碳库中所占比例(52%)远高于微生物生物量碳(3%~6%);3种菌种对土壤养分含量的提升效应表现为:RI > CE > FM。相较于对照,RI 菌种对植物可利用性氮、微生物生物量碳、总有机碳及植物可利用性磷的提升率分别为161%、127%、110%及97%;对ROC变化具有较大贡献的土壤环境因子分别为AM真菌侵染率(96%)、植物可利用性氮(94%)、微生物生物量碳(85%)、总有机碳(78%)及植物可利用性磷(72%)。【结论】AM真菌与尼泊尔桤木共生,显著驱动石漠化土壤碳库与养分含量变化并促进植物生长,进而增加石漠化土壤易氧化碳的积累。研究结果有助于理解石漠化地区植物生长、土壤恢复及活性有机碳沉积的微生物学调控机制。     

东寨港红树林丛枝菌根真菌侵染和孢子密度及其影响因子研究

丛枝菌根(AM)结构存在于多种红树植物根内,但目前对于红树林中AM真菌的研究还未能完全揭示其存在的原因和状况。研究通过对海南东寨港红树林中不同潮位16种红树植物根内AM侵染情况、AM真菌孢子密度以及土壤理化因子开展调查,以了解东寨港红树林中AM真菌的存在状况及其影响因素。结果表明12种红树植物与AM真菌存在共生关系,其中海漆(Excoecaria agallocha)、许树(Clerodendrum inerme)、水黄皮(Pongamia pinnata)和黄槿(Hibiscus tiliaceus)侵染率较高,均高于55%;16种植物根际土壤中均检测到AM真菌孢子,每25 g土壤的平均孢子密度为(25.7±2.7)个。方差分析表明,不同宿主植物根内AM侵染情况差异显著;潮位对AM侵染率和孢子密度均具有显著影响,表现为高潮位显著高于中、低潮位,而中低潮位间无显著差异。线性回归分析表明土壤含水率、电导率和有效磷含量可能是影响东寨港红树林生境中AM侵染率和孢子密度的主要土壤因子。     

金银花根围AM真菌分布与土壤碳氮关系

为充分利用AM真菌资源促进中药材生长,2010年7-12月,连续从河北省安国市药材种植基地采集金银花(Lonicera japonica)根围土壤样品,系统研究了AM真菌时空分布及其与土壤碳氮含量的相关性.结果表明,金银花能与AM真菌形成良好共生关系;AM真菌最大孢子密度为59.02 g-1,最高定殖率为97％;孢子密...     

Inhibition of shoot branching by new terpenoid plant hormones

Shoot branching is a major determinant of plant architecture and is highly regulated by endogenous and environmental cues. Two classes of hormones, auxin and cytokinin, have long been known to have an important involvement in controlling shoot branching. Previous studies using a series of mutants with enhanced shoot branching suggested the existence of a third class of hormone(s) that is derived from carotenoids, but its chemical identity has been unknown. Here we show that levels of strigolactones, a group of terpenoid lactones, are significantly reduced in some of the branching mutants. Furthermore, application of strigolactones inhibits shoot branching in these mutants. Strigolactones were previously found in root exudates acting as communication chemicals with parasitic weeds and symbiotic arbuscular mycorrhizal fungi. Thus, we propose that strigolactones act as a new hormone class-or their biosynthetic precursors-in regulating above-ground plant architecture, and also have a function in underground communication with other neighbouring organisms.     

三种水份条件下湿生植物Lythrum salicaria L.的菌根状况及其对植物生长的影响

陆生植物的菌根已被广泛研究,对湿生植物菌根的研究却很少.本研究的目的是检测在水淹、潮湿和干旱条件下水生植物L.salicaria的菌根合成状况及其对植物生长的影响.在三种水份条件下,L.salicaria与来源于不同地点(干旱和水淹)的两种接种物(F2和F1)均可形成菌根,水份条件和接种物种类都显著地影响菌根的合成水平(P《0.05).F1和F2接种物与植物合成菌根的水平均在水淹处理中最低. F1接种物与L.salicaria合成菌根的水平在潮湿和干旱处理中显著地高于水淹处理(P《0.05),并且趋势是:潮湿》干旱》水淹处理.F2接种物与L.salicaria合成菌根的水平在三种水份处理中没有显著区别,但仍然可观察到一个趋势:干旱》潮湿》水淹处理.在水淹和干旱处理中,菌根对植物地上部分的生长和根的生物量积累没有影响.在潮湿处理中被F1接种的植株与被F2接种的植株与对照植株相比,地上部分干重、根鲜重显著地低;地上部分鲜/干重比、地上部分/根鲜重比显著地高.水份条件显著地影响植物的生长行为.     

Organization of genetic variation in individuals of arbuscular mycorrhizal fungi

Arbuscular mycorrhizal (AM) fungi (Glomeromycota) are thought to be the oldest group of asexual multicellular organisms. They colonize the roots of most land plants, where they facilitate mineral uptake from the soil in exchange for plant-assimilated carbon. Cells of AM fungi contain hundreds of nuclei. Unusual polymorphism of ribosomal DNA observed in individual spores of AM fungi inspired a hypothesis that heterokaryosis--that is, the coexistence of many dissimilar nuclei in cells--occurs throughout the AM fungal life history. Here we report a genetic approach to test the hypothesis of heterokaryosis in AM fungi. Our study of the transmission of polymorphic genetic markers in natural isolates of Glomus etunicatum, coupled with direct amplification of rDNA from microdissected nuclei by polymerase chain reaction, supports the alternative hypothesis of homokaryosis, in which nuclei populating AM fungal individuals are genetically uniform. Intrasporal rDNA polymorphism contained in each nucleus signals a relaxation of concerted evolution, a recombination-driven process that is responsible for homogenizing rDNA repeats. Polyploid organization of glomeromycotan genomes could accommodate intranuclear rDNA polymorphism and buffer these apparently asexual organisms against the effects of accumulating mutations.     

不同接种物对分离丛枝菌根真菌的影响

在实验室条件下,用野外采集的植物根际土壤和野生葱属植物根段作为接种体接种三叶草(TrifoliumrepensL.).观察三叶草菌根侵染率,统计其根际土壤中丛枝菌根真菌的孢子密度和种类、并与原始接种土壤比较.结果表明:用不同植物群落的土样作为接种物,三叶草菌根侵染率和根际AMF种类和密度有很大差异,但菌根真菌种类数目较多;用不同种葱属植物根段接种,三叶草菌根的侵染率差别明显,根际土壤孢子的种类数目较少但菌根真菌有很高的相似性.     

AM真菌克服作物连作障碍的潜力

连作障碍是农业生产和设施栽培模式下的主要限制因子之一。连作障碍主要与土壤微生物区系的变化、土壤理化性状的变化和根系分泌物的自毒作用三方面密切相关。已有证据表明,丛枝菌根(AM)真菌在一定程度上能够克服连作障碍。这很可能与AM真菌减少有害生物数量,改变土壤微生物区系、改善土壤理化性状、提高植物防御性酶活性、改变根系分泌物的种类与数量等生理功能有关。正确合理评价AM真菌克服作物连作障碍的潜力,可为连作障碍提供新的解决途径,并为建立和发展新的生物技术提供依据。     

AM宿主植物金银花对喀斯特干湿交替的光合响应

以盆栽的方式,探究了接种丛枝菌根(AM)真菌后金银花在西南岩溶地干湿交替的变水环境下光合响应的变化,期望为喀斯特地区生态环境的恢复提供思路.结果表明:在适宜水分条件下,接种AM真菌促进了金银花的净光合速率(Pn),蒸腾速率(Tr),气孔导度(Gs)和气孔密度.在短期干旱条件下,接种AM真菌对Pn和Tr均没有明显影响;然而干旱持续时间加长后,Pn有显著提高,Gs受到抑制,Tr相比未接种组反而降低,水分利用效率(WUEi)有所上升.在短期干湿交替下,接种AM真菌后各项光合指标均没有显著提高,反而金银花植株的Pn和Tr被抑制,长期干湿交替的WUEi虽然提高但是以牺牲植物的光合作用为代价.总之,接种真菌对适宜水分下金银花光合能力的促进最为明显,干旱下有一定促进能力,但是在反复的干湿交替变水下,接种植株的光合作用没有得到显著促进.