叶际微生物及其与外界互作的研究进展

叶际微生物及其生存环境共同形成了一个复杂的生态系统。建立在纯种分离和纯培养技术基础之上的传统研究方法只能了解其中部分叶际微生物,但对物种组成、种群结构和生态学作用等方面的认识都比较片面。近年来随着分子生物学和生物信息学的进步,人们对叶际微生物总群落的分析逐渐揭示了叶际微生物组成的多样性及其特点,以及与外界互相作用的复杂性。研究表明,植物种类、地理位置和季节差异等都不同程度地影响着叶际微生物群落的构成。本文综述了近年来国内外叶际微生物群落结构组成及其与外界互作方面的研究进展,有利于加深对叶际微生物的了解,也有助于深入理解叶际微生物与植物生长和植物病虫害防治的关联关系。     

叶际微生物与植物健康研究进展

叶际微生物与植物有着密切的联系,对宿主生物多样性的维持、群落的稳定和生态系统的功能具有重要意义。随着分子生物学技术的快速发展,叶际微生物与植物的关系及对宿主健康的影响是近年来热点领域之一。基于明确定义的叶际概念,综述了叶际微生物群落结构、驱动因素及其与植物健康的关系,强调了叶际微生物的机遇性以及调控群落驱动因素在未来叶际微生物研究中的重要地位。加深对叶际微生物的认知,有利于实现农业微生物的功能最大化,以期为提高植物产量提供参考。     

北京龙形水系三种沉水植物根际及叶际微生物群落特征

水生植物及植物表面附着微生物在人工湿地水体净化过程中发挥着重要的作用。以北京奥林匹克公园龙形水系为研究对象,通过高通量测序技术,对其底泥、水体及3种沉水植物——苦草Vallisneria natans、狐尾藻Myriophyllum verticillatum、龙须眼子菜Potamogeton pectinatus——的根际及叶际微生物群落的结构及功能进行了研究。结果表明,微生物多样性从高到低分别为底泥样品、植物根际样品、植物叶际样品和水体样品,植物叶际微生物种类要显著高于水体中微生物种类。LEfSe分析结果显示不同生境富集不同的微生物类群,其中底泥主要富集厌氧微生物类群,水体及植物叶际主要富集好氧微生物类群,植物根际则两者兼具。功能预测结果显示植物叶际样品的反硝化标志基因丰度要高于根际样品及底泥和水体样品,且狐尾藻和龙须眼子菜叶际样品反硝化标志基因丰度要高于苦草叶际样品。本研究可以为人工湿地构建时对沉水植物及功能微生物的选择提供指导意义。     

植物叶际固氮菌研究进展

固氮菌广泛存在于植物叶际,能固定空气中的氮气,满足自身或植物的部分氮素需求。基于纯种分离和培养的传统生物学方法已经研究了部分叶际固氮菌的特性,但对叶际固氮菌的物种组成、群落结构及生态功能等方面的认识还非常有限。随着分子生物学技术的发展、微生物分子生态学研究方法的逐步成熟,人们对叶际微生物的多样性和生态功能的研究越来越深入。叶际固氮菌具有丰富的多样性,温度、湿度、光照等环境因素及植物种类和微生物互作均会影响叶际固氮菌的组成。不同于根际固氮菌,叶际固氮菌具有非专一性,且不易受化肥的影响,其在农业生产上已经表现出潜在的应用价值。为此,本文综述了农作物、森林、海域生态系统中叶际固氮菌的群落结构组成及生态功能,以及外界因素对叶际固氮系统的影响。     

叶际微生物对马尾松凋落针叶分解的影响

叶际微生物作为最先定殖在凋落叶上的微生物类群,可能直接参与凋落叶的分解。为验证此猜想,该研究通过扩增子高通量测序技术和室内分解实验,探究了马尾松(Pinusmassoniana)叶际微生物多样性及叶际微生物对马尾松凋落物的分解影响。结果表明:(1)马尾松的叶际存在着丰富而多样的微生物群体,针叶在凋亡后,叶际微生物群落发生变化。成熟针叶、凋落针叶、分解层针叶共有大量可操作分类单元(OTUs)。(2)马尾松针叶分解过程可分为两个阶段:快速分解期(前8个月)和缓慢分解期(8个月以后)。衰亡针叶(刚凋落但未接触土壤)叶际微生物可直接参与马尾松凋落针叶分解,且分解速率表现为叶际微生物+土壤微生物处理>叶际微生物处理>土壤微生物处理。在马尾松针叶分解过程中叶际微生物与土壤微生物存在协同作用。(3)凋落针叶分解速率与木质素和纤维素分解速率呈极显著正相关关系,但与木质素和纤维素分解酶活性无显著相关关系。木质素分解酶——多酚氧化酶与过氧化物酶活性极显著负相关,纤维素分解酶——β葡萄糖苷酶活性与纤维二糖苷酶活性则呈极显著正相关关系。综上,该研究结果表明叶际微生物可直接参与凋落针叶的分解,且其对...     

长叶红砂叶际细菌和真菌群落对季节变化的响应特征

【背景】植物叶际(phyllosphere)定殖着丰富多样的微生物,叶际微生物通过发挥特定功能在逆境下生存,影响寄主植物的生理生态特性并受环境异质性的影响。【目的】植物叶际微生物群落是动态的,认识季节更替对植物叶际微生物群落结构的影响,对于加深对植物-微生物-环境相互作用的理解具有积极意义。【方法】以鄂尔多斯荒漠草原泌盐盐生植物长叶红砂为研究对象,分别测定春季、秋季植物叶片表面理化特性,并结合叶际细菌、真菌高通量测序结果进行综合分析。【结果】长叶红砂冠下土壤含水率、pH、电导率等指标在季节更替下存在显著差异,叶片表面Na+、K+和电导率值存在显著差异;进一步分析发现,叶际细菌分类操作单元(operational taxonomic unit,OTU)、Shannon、Chao1和ACE (abundance-based coverage estimator)指数与土壤和叶片表面盐分含量呈正相关;春季叶际蓝细菌门和拟杆菌门保持了较高的相对丰度,而秋季叶际变形菌门、放线菌门、子囊菌门的相对丰度则高于春季;其中,叶际Bradyrhizobium、Novosphingobium和Edaphob...     

模拟增温及分解界面对茭草凋落物分解速率及叶际微生物结构和功能的影响

挺水植物的凋落物是湿地生态系统物质循环的重要组成部分, 阐明气候变暖以及生境差异对湿地挺水植物凋落物分解过程及叶际微生物的影响对揭示湿地生态系统关键物质循环过程具有重要意义。该研究以滇西北高原典型湿地优势挺水植物茭草(Zizania latifolia)为研究对象, 采用凋落物袋法研究了茭草在模拟增温(1.5-2.0 ℃)及不同生境(大气界面、水界面与土界面)下的质量残留率和叶际微生物数量、结构组成与功能代谢特征。研究发现: 模拟气候变暖及生境差异均显著影响凋落物的分解速率。经过一年的分解, 凋落物在模拟增温环境下的质量残留率为66.4%, 对照组的质量残留率为77.7%, 增温组分解常数(k)值是对照组的1.64倍, 而凋落物在水界面与土界面的质量残留率为42.2%和25.3%, 其k值分别为大气界面的3.63和5.25倍, 生境差异是影响湿地挺水植物凋落物分解速率的关键因素。模拟增温主要改变了凋落物叶际微生物的群落组成特征, 而生境变化主要影响叶际微生物的绝对数量、微生物多样性、群落结构组成以及功能代谢活性。处于土界面的凋落物叶际微生物具有最高的群落功能代谢活性及醇类碳源利用程度。不同处理之间的植物叶际微生物特征与凋落物分解速率具有较好的一致性, 为揭示湿地植物凋落物分解快慢的微生物驱动机制提供了重要的理论依据。     

叶际微生物诱发气孔免疫的机制及其应用前景北大核心CSCD

植物叶际微生物通过自身代谢活动影响植物功能的正常运行。而植物则可感应叶际微生物的存在,在诱发气孔免疫关闭以抵御病原菌入侵的同时,也降低了植物蒸腾作用,提高了其水分利用效率。对气孔免疫相关理论的研究有利于开发新型生物抗蒸腾剂,并对开发节水农业新技术具有重要意义。该文综述了叶际微生物与植物间的互作关系、气孔免疫及其免疫机制等方面的研究进展,并探讨了相关机制在节水农业方面的应用前景和重点研究方向。     

拟南芥与叶际微生物组的互作遗传机制——基于网络作图理论

叶际微生物组对植物的生长发育至关重要，但植物与其定殖微生物组相互作用机制尚不明确。目前植物与微生物互作研究多集中于根际微生物组，对叶际微生物组的研究较少，且这些研究未能从微生物互作的角度探究植物与微生物的相互作用机理。基于网络作图理论，将拟南芥基因组SNP （Single Nucleotide Polymorphisms）分子标记数据与微生物组网络特征值相关联，挖掘影响叶际微生物组网络结构的枢纽基因，以探究拟南芥塑造叶际微生物组网络结构的遗传机制。通过对188株拟南芥及其叶际微生物组数据的分析，识别出四种关系下的中心节点微生物，筛选到622个显著SNP位点。进一步构建了贝叶斯遗传网络，获得26个枢纽基因，这些基因可能参与了植物抗病、激素分泌和生长发育相关的分子途径。本研究从全基因组角度探究植物调控自身微生物组的遗传机制，揭示植物与微生物组如何互作促进植物健康，将为精准分子育种提供理论基础和遗传资源，并为合成菌群用于创制新型菌剂提供数据支持，具有重要的科学意义和应用价值。     

不同生境中桂花和夹竹桃叶际细菌的群落结构

植物叶际微生物多样性是目前植物-微生物关系研究的热点之一,但影响叶际微生物群落结构的主要因素目前还存在很大争议.本研究以生长在3处生境的桂花和夹竹桃为对象,基于高通量测序技术,分析2种植物叶际细菌的群落结构,探讨影响植物叶际细菌群落结构的主要因素.结果表明:来自3处生境的2种植物叶际细菌多样性无显著差异,构成叶际细菌群落的优势门主要包括放线菌门、拟杆菌门、衣原体门、蓝细菌门、厚壁菌门和变形菌门,优势属主要包括甲基杆菌属、鞘氨醇单胞菌属、薄层杆属、Polaromonas和无毛螺旋体属.植物种类、生境及二者的交互作用均能显著影响叶际细菌群落结构,其中生境的影响最大.     

Composition of the Phyllospheric Microbial Populations on Vegetable Plants with Different Glucosinolate and Carotenoid Compositions

The plant phyllosphere is intensely colonized by a complex and highly diverse microbial population and shows pronounced plant-species-specific differences. The mechanisms and influencing factors determining whether and in which density microorganisms colonize plant phyllosphere tissues are not yet fully understood. One of the key influencing factors is thought to be phytochemical concentration and composition. Therefore, correlations between various concentrations of individual glucosinolates and carotenoids in four different plant species-Brassica juncea, Brassica campestris, Cichorium endivia, and Spinacea oleracea-and the phyllospheric bacterial population size associated with the aerial parts of the same plants were analyzed. The concentration of various individual glucosinolates and carotenoids were measured using high-performance liquid chromatography. The phyllospheric bacterial population size including both nonculturable and culturable organisms was assessed using quantitative real-time polymerase chain reaction, and the physiological profile of the culturable microbial community was analyzed using the Biolog system. Results show significant differences between plant species in both concentration and composition of secondary metabolites, bacterial population size, and microbial community composition in three consecutively performed experiments. An interesting and underlying trend was that bacterial density was positively correlated to concentrations of beta-carotene in the plant phyllosphere of the four plant species examined. Likewise, the alkenyl glucosinolates, 2-propenyl, 3-butenyl, and 4-pentenyl, concentrations were positively correlated to the bacterial population density, whereas the aromatic glucosinolate 2-phenylethyl showed a negative correlation to the phyllospheric bacterial population size. Thus, we report for the first time the relationship between individual glucosinolate and carotenoid concentrations and the phyllospheric bacterial population size of nonculturable and culturable organisms and the phyllospheric microbial physiological profiles.     

Summer season and long-term drought increase the richness of bacteria and fungi in the foliar phyllosphere of Quercus ilex in a mixed Mediterranean forest

We explored the changes in richness, diversity and evenness of epiphytic (on the leaf surface) and endophytic (within leaf tissues) bacteria and fungi in the foliar phyllosphere of Quercus ilex, the dominant tree species of Mediterranean forests. Bacteria and fungi were assessed during ontogenic development of the leaves, from the wet spring to the dry summer season in control plots and in plots subjected to drought conditions mimicking those projected for future decades. Our aim was to monitor succession in microbiota during the colonisation of plant leaves and its response to climate change. Ontogeny and seasonality exerted a strong influence on richness and diversity of the microbial phyllosphere community, which decreased in summer in the whole leaf and increased in summer in the epiphytic phyllosphere. Drought precluded the decrease in whole leaf phyllosphere diversity and increased the rise in the epiphytic phyllosphere. Both whole leaf bacterial and fungal richness decreased with the decrease in physiological activity and productivity of the summer season in control trees. As expected, the richness of epiphytic bacteria and fungi increased in summer after increasing time of colonisation. Under summer dry conditions, there was a positive relationship between TRF (terminal restriction fragments) richness and drought, both for whole leaf and epiphytic phyllosphere, and especially for fungal communities. These results demonstrate that changes in climate are likely to significantly alter microbial abundance and composition of the phyllosphere. Given the diverse functions and large number of phyllospheric microbes, the potential functional implications of such community shifts warrant exploration.     

种子微生物生态学研究进展

植物种子微生物生态学是研究与种子相联合的微生物的组成﹑功能﹑演替、它们之间关系及其与宿主之间相互关系的科学。种子中蕴含着丰富的微生物资源,它们对种子以及植物的健康具有重要的影响。不同种类植物种子联合的微生物群落由于受到种子本身及外界环境因素的影响而有所差异。论述了种子微生物生态学的概念、主要研究方法、种子微生物生态系统中的微生物种类、相关影响因素,以及种子微生物生态学研究的发展方向。种子微生物生态学的研究对生产实践有重要意义,同时也将丰富种子生物学的内容,对种子科学的发展起到促进作用。     

The influence of air pollution on the phyllosphere microflora composition of Tillandsia leaves (Bromeliaceae)

The effect of air pollution on total phyllospheric microflora from two species of the epiphytic neotropical genus Tillandsia (Bromeliaceae) was studied by comparing unpolluted plants living in a forest (Escazú, San José) with polluted ones from an urban site of Costa Rica (San José city). Dilutions of homogenized leaf samples were plated on media suitable for each microbial group. For each microorganism group, total counts were performed and purified strains of randomly chosen colonies were identified. There was a global reduction in the number of living microorganisms due to pollution effects, especially yeasts and bacteria, while nitrogen-fixing microorganisms and fungi were less affected. Our results showed that the phyllosphere microflora of Tillandsia plants living in a tropical urban environment changes in terms of number and species composition of yeasts and bacteria with respect to plants living in unpolluted environment.     

青藏高原垫状点地梅叶际及内生可培养微生物多样性

【背景】垫状点地梅作为青藏高原最具代表性的垫状植物,其叶际和内生微生物对适应极端环境有重要意义,同时也是一种独特的资源。【目的】探究垫状点地梅叶际和叶内可培养微生物多样性,以及不同生存状态个体之间的微生物差异。【方法】采用纯培养方法分离和纯化3个不同地区垫状点地梅叶际和叶内的细菌、酵母菌和丝状真菌,并用16S rRNA基因和ITS区域序列进行分析鉴定。【结果】最终得到叶际微生物350株,鉴定为22属49种,优势种为Penicillium sajarovii;内生微生物274株,鉴定为19属45种,优势种为Bacillusmycoides;两者的优势属均为Penicillium。垫状点地梅叶际和叶内之间及不同生存状态个体之间微生物的α多样性大多无显著差异,各群落间的成员也有重叠,但物种组成存在显著的空间异质性。【结论】垫状点地梅叶际和叶内有着丰富的可培养微生物资源,来源于不同生存状态的个体或不同部位的微生物物种组成差别较大,微生物对不同环境的选择偏好形成了不同的群落模式。但这些不同来源的微生物群落之间同样存在高比例的共有菌株,这些共有菌株的异养方式和生态位并不固定,可兼共生和腐生生存,生...     

Characterization of xylanolytic bacteria present in the bract phyllosphere of the date palm Phoenix dactylifera

AIMS: Despite the interest of phyllosphere microbiology, no studies have addressed the bacteria present in bract phyllosphere, an ecosystem that has special characteristics in palm trees because the dry bracts remain on the plant until pruning and may contain polymer-degrading bacteria involved in plant degradation. Therefore, the aim of this work was to characterize xylanolytic bacteria isolated from palm bract phyllosphere. METHODS AND RESULTS: Twelve xylanolytic strains were isolated and characterized by phenotypic features and complete sequencing of 16S rRNA gene. The results showed that the isolates were phenotypically and genotypically diverse. Gram-positive isolates were classified into genus Paenibacillus some of them belonging to hitherto undescribed species of this genus. Gram-negative isolates were classified into genera Pseudomonas and Acinetobacter. CONCLUSIONS: The results of this work confirm the complexity of the bacterial populations present in phyllospheric ecosystems and suggest that bacteria involved in plant degradation are present at the early degradation steps of this process in dry palm tree bracts. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study on bract phyllospheric bacteria able to hydrolyse vegetal polymers and offers a new perspective in the search of unexplored sources of xylanase-producing strains.     

Population and community level approaches for analysing microbial diversity in natural environments

The enormous diversity available at the microbial level is just beginning to be realized. The richness of diversity amongst the bacteria that have been described so far is between 2 and 3000, whereas estimates indicates that millions of microorganisms still remains to be discovered. Microbiologists have realized that there are at least a dozen major evolutionary groups of the microbial life forms on earth (bacteria, fungi, algae and protozoa) that are even more diverse than the better known animal and plant kingdom. Indeed, we can state that microorganisms dominate the tree of life. Microorganisms have inhabited Earth for more than 3.7 billion years, whereas plants and animals have evolved rather recently in Earth's history. Possible reports of evidence for microbial life on Mars is also consistent with the concept that microorganisms precede plants and animals on Earth. The applications of molecular-phylogenetic techniques have provided the tools for studying natural microbial communities, including those that we are not able to grow in the laboratory. The utilization of these techniques has resulted in the discovery of many new evolutionary lineages, some of them only distantly related to known organisms. Here I discuss some environmental factors controlling bacterial diversity in different environments and the utility of modern methods developed for describing this diversity.     

海拔梯度变化对中亚热带黄山松土壤微生物生物量和群落结构的影响

全球变暖对陆地生态系统造成一系列生态问题,使这些问题将随着全球平均气温的升高而进一步加剧。海拔梯度变化是研究气候变暖对陆地生态系统影响的一种重要手段。目前为止利用海拔梯度对微生物影响的研究尚未定论,其主要原因是忽略了植被类型的影响。因此,以中亚热带戴云山的3个海拔(1300、1450、1600 m)的黄山松(Pinus taiwanensis)林为研究对象,探究沿海拔梯度的变化,森林土壤微生物生物量和微生物群落结构的响应变化。结果表明:土壤碳氮磷养分(SOC、TN、TP)、微生物生物量氮(MBN)、微生物生物量磷(MBP)和丛枝菌根真菌(AMF)、革兰氏阴性菌(GN)、真菌(Fungi)、总磷脂脂肪酸(T_(PLFA)),细菌∶真菌(F∶B)均随海拔升高显著下降,而革兰氏阳性菌∶革兰氏阴性菌(GP∶GN)随海拔升高呈相反的趋势。冗余分析(RDA)表明,温度(T)和可溶性有机氮(DON)是影响微生物群落结构的最重要的环境因子。研究表明:与1600 m海拔相比,1300 m海拔温度较高,土壤有机质矿化作用较强,土壤速效养分及微生物生物量随之增加,从而提高(Fungi)、细菌(Bacteria)等。因此,未来气候变暖将通过改变土壤碳氮磷养分来影响本区域微生物群落组成结构。这对进一步深入了解气候变化对山地生态系统土壤养分循环过程具有重要意义。     

Role of phyllosphere fungi of forest trees in the development of decomposer fungal communities and decomposition processes of leaf litter

The ecology of endophytic and epiphytic phyllosphere fungi of forest trees is reviewed with special emphasis on the development of decomposer fungal communities and decomposition processes of leaf litter. A total of 41 genera of phyllosphere fungi have been reported to occur on leaf litter of tree species in 19 genera. The relative proportion of phyllosphere fungi in decomposer fungal communities ranges from 2% to 100%. Phyllosphere fungi generally disappear in the early stages of decomposition, although a few species persist until the late stages. Phyllosphere fungi have the ability to utilize various organic compounds as carbon sources, and the marked decomposing ability is associated with ligninolytic activity. The role of phyllosphere fungi in the decomposition of soluble components during the early stages is relatively small in spite of their frequent occurrence. Recently, the roles of phyllosphere fungi in the decomposition of structural components have been documented with reference to lignin and cellulose decomposition, nutrient dynamics, and accumulation and decomposition of soil organic matter. It is clear from this review that several of the common phyllosphere fungi of forest trees are primarily saprobic, being specifically adapted to colonize and utilize dead host tissue, and that some phyllosphere fungi with marked abilities to decompose litter components play important roles in decomposition of structural components, nutrient dynamics, and soil organic matter accumulation.