豆科植物-根瘤菌共生固氮的免疫调控机制

在长期进化中,根瘤菌与豆科植物形成一种独特的互惠共生关系——共生固氮。根瘤菌-豆科植物共生互作与病原细菌激发植物病原反应极为相似,然而根瘤菌的入侵和定殖并没有激发宿主豆科植物过度的防御反应,植物也进化出特殊的共生信号转导和根瘤发育途径来"邀请"根瘤菌的入侵和定殖。此外,植物防御反应也很大程度上调控根瘤菌与豆科植物共生的宿主特异性。越来越多的研究表明,植物防御反应在调控根瘤菌匹配识别、入侵、定殖以及类菌体发育等方面起关键调控作用。从植物免疫反应角度综述了根瘤菌与豆科植物共生互作的最新进展,通过与病原菌-植物互作的病原反应对比,论述了根瘤中植物感知微生物相关分子模式(MAMP,Microbe-Associated Molecular Patterns)和效应蛋白引起的免疫反应的调控机制。     

植物病原菌效应子的研究进展

病原菌为了成功侵入并在寄主植物中繁殖,会分泌效应子作为入侵武器.不同病原菌的效应子具有一定的共性和异性.开展植物病原菌效应子的系统鉴定,深入揭示效应子对病原菌侵入和在植物发病中的作用以及解析效应子与植物抗病基因的互作,可为研究病原菌的致病机制及其与植物的互作提供重要的研究线索,在植物病理和抗病遗传育种研究中也具有重要理论价值和实践意义.近年来,随着测序技术的不断发展,基因组学和转录组学在植物抗病研究中的应用也日益广泛,其研究结果可为鉴定病原物致病基因、植物抗病基因、阐明病原菌与植物互作的分子机制提供重要信息.本文根据近年来植物包括树木中病原物效应子的研究进展,对效应子的特点、鉴定方法、功能及宿主抗病机理等进行了综述和比较,重点阐述了效应子的鉴定、致病功能及与植物抗病基因的分子互作和调控,并对效应子在植物抗病中的应用及其研究前景进行了展望.     

植物内生菌群落组成及其功能研究进展

植物内生菌是栖居在植物组织内部、以宿主植物代谢物为营养物质的一类微生物。植物内生菌种类丰富,并通过不同的方式定殖在植物体内,与宿主植物互利共生。植物内生菌促进植物的生长发育,通过增强营养物质吸收、与病原菌竞争生态位、在代谢过程中产生抗菌物质以及诱导宿主植物产生抗性等机制,提高宿主植物的抗逆性,并且其群落的组成因生态环境、寄主植物不同部位、生长周期的不同发育阶段而显著不同。人们利用传统组织分离法、高通量测序法以及人工重组生物群落等研究方法对植物与内生菌相互作用进行了深入的研究。本文对植物内生菌群落的组成、内生菌对宿主植物影响的功能研究以及人工内生菌群的构建进行重点阐述,以期为植物内生菌的开发和利用奠定坚实基础。     

植物内生菌定殖检测技术及其应用

自然环境中内生菌定殖于植物体内,对宿主植物产生多种有益效应,但是内生菌定殖情况难以检测,相关研究不够深入系统。目前在该领域使用较为广泛的检测技术包括：荧光标记、抗生素标记、荧光定量PCR和高通量测序等。内生菌通过孔隙伤口和降解细胞壁等方式侵染植物,通过种子垂直传递核心内生菌。对植物内生菌定殖的侵染方式、定殖方法和检测技术进行了归纳和整理,介绍了内生菌多种侵染和迁移途径,总结了目前内生菌定殖在生物防治、促进植物生长和污染修复等方面的功能,综述了多种检测方式在应用中的特点,以期为内生菌定殖植物的相关研究及其应用提供参考依据。     

丛枝霸王（Zygophyllum dumosum）根际AM真菌生态学研究

AM真菌是一类广泛分布的土壤真菌,与宿主植物形成共生结构后,对于植物生长和植被恢复有多种有益的生理学和生态学作用。1999年11月至2000年10月,通过每月分别从0－10cm和10－20cm土层采集根际土样,对以色列荒漠地区丛枝霸王（Zygophyllum dumosum Boiss）根际AM真菌进行了系统的生态学研究。AM真菌的分布和定殖与季节变化和采样浓度密切相关。菌根真菌的最高定殖率并不伴随有最大的孢子密度,最高的定殖率发生在1999年11月,而最大的孢子密度则出现在2000年9月。10－20cm土层中的菌根真菌定殖率和孢子密度明显高于0－10cm土层。土壤温度与所有菌根结构的定殖率呈正相关,土壤有机质含量与泡囊和丛枝定殖率呈正相关,而土壤总可溶性氮对泡囊和丛枝定殖有显著正效应,对孢子密度有显著负效应。结果建议,孢子密度、泡囊和丛枝定殖程度可作为检测荒漠土壤状况的生态指标。研究应用于我国特别是西部荒漠地区的植被建立和恢复,可望发挥重任作用：（1）AM真菌能与绝大多数高奶系形成共生联合体,促进根系对土壤矿质营养和水分的吸收,提高植物对干旱、高温、盐碱、根部病害等的抗性,提高逆境条件下植物的成活率,深入研究荒漠生态系统中AM真菌动态分布,以及筛选优势AM菌种和人工接种,进行菌根化育苗,为植被建立和恢复提供优质苗木;（2）菌根的丛枝定殖时间短,主要发生在幼根,泡囊定殖时间长,主要发生在老根,而AM真菌的生长发育和繁殖所需的碳水化合物来自植物根系的分泌活动,所以,通过检测不同时期菌根各种结构的定殖程度和孢子的丰富度,可以获得宿主植物根系的生长状况,进而对土壤环境作出科学的评估。     

效应子及其与小麦条锈菌致病性的关系

植物病原菌和寄主植物竞争进化过程中分泌称为效应子(Effectors)的复杂的分子,其逃避或抑制寄主的免疫反应,干扰寄主的各种生理过程,从而有利于提高病原菌寄生适合度、定殖能力和传播能力。利用基因组序列和转录组测序数据预测小麦条锈菌中含有两千多个候选效应子,但由于小麦条锈菌缺乏有效的遗传转化体系,目前对其效应子功能的研究进展缓慢。在介绍效应子分类及其特征基础上,结合可运用于小麦条锈菌效应子功能研究的方法,从分子水平上阐释小麦条锈菌与寄主植物互作取得的进展,关注效应子靶定的寄主亚细胞成分及其与致病性的关系。这些进展有利于深入认识小麦条锈菌效应子的生物学功能,并为深入探讨小麦条锈菌的致病机制,制定新的防治策略提供方向。     

玉米瘤黑粉菌的遗传交配型

玉米瘤黑粉病是玉米的一种重要病害,普遍分布于世界各玉米产区,我国各地也有不同程度的发生,主要症状是在玉米的茎、叶、雄花、雌穗等部位形成肿瘤［１］。其病原菌为玉米瘤黑粉菌（Ｕｓｔｉｌａｌｇｏ　ｍａｙｄｉｓ）,属于担子菌亚门,异宗配合。在玉米瘤黑粉菌的生活史中,有两种不同形态的细胞,即单倍体细胞（担孢子）和双核菌丝体。单倍体细胞没有致病性,在特定培养基上芽殖产生“酵母”状菌落。不同遗传型的单倍体细胞融合形成双核菌丝,双核菌丝能在寄主植物体内迅速发育,刺激寄主组织形成肿瘤,并继而经过细胞核融合,产生双…     

金黄色葡萄球菌壁磷壁酸致病与免疫的分子机制研究进展

金黄色葡萄球菌(Staphylococcus aureus)壁磷壁酸(wall teichoic acids, WTAs)是多元醇经由磷酸二酯键共价连接组成的细胞壁表面阴离子糖类聚合物,参与调节细胞壁的稳态并介导细菌毒力。金黄色葡萄球菌WTAs与宿主细胞表面特定的受体结合,可诱导天然免疫和获得性免疫应答。此外,金黄色葡萄球菌WTAs还参与调控毒力基因的表达,有助于细菌的定殖感染,在基因工程靶标治疗和噬菌体药物治疗方面具有广泛的应用前景。本文对金黄色葡萄球菌WTAs的合成进行了概述,综述了WTAs对宿主免疫应答的调控作用,以及在细菌对宿主侵袭与定殖中的致病机制,并归纳WTAs的耐药分子机制和作为药物治疗靶标的研究现状。这些研究为揭示WTAs的致病与免疫分子机制提供研究思路,为预防和治疗金黄色葡萄球菌的感染提供新的策略。     

干旱胁迫下内生真菌感染对羽茅的生理生态影响

本研究在田间环境下对感染和未感染内生真菌的天然宿主羽茅（Achnatherum sibiricum (L. ) Keng）进行了干旱胁迫实验,结果发现在干旱胁迫下,内生真菌感染对宿主植物的营养生长、生物量累积和叶绿素含量都没有显著影响,但对宿主植株光系统II光化学效率（Fv/Fm）的维持产生了有利效应。同时,内生真菌感染缓解了宿主植物细胞膜的旱害程度,表现在与未感染植株相比,感染植株的丙二醛（MDA）含量显著降低,但内生真菌的感染并未促使宿主植物体内保护酶如超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活性的增加,只是显著增加了类胡萝卜素的含量。因此我们推测在羽茅中,内生真菌对宿主植物的保护作用可能更多的体现在非酶系统上。     

SPINDLY在壳寡糖诱导拟南芥抗丁香假单胞菌中的功能

为了探讨拟南芥O-岩藻糖基转移酶(SPINDLY)在病原体相关分子模式诱导抗性中的作用,该研究以SPINDLY缺失拟南芥突变体spy-3为实验材料,从叶片表型、病情指数、病菌定殖量以及丁香假单胞菌(Pst DC3000)关键基因的表达水平等指标,系统考察了SPINDLY在壳寡糖诱导拟南芥抗Pst DC3000中的功能。结果显示:(1)spy-3突变体比野生型更易被Pst DC3000侵染。(2)与病菌侵染组相比,壳寡糖预处理明显缓解植株叶片黄化现象,显著降低Pst DC3000的定殖量。(3)壳寡糖预处理的spy-3植株中水杨酸和茉莉酸途径相关基因的表达量及水杨酸和茉莉酸含量均较病菌侵染组明显升高。(4)壳寡糖在spy-3中的诱抗效果与野生型相比无明显差别。研究表明,SPINDLY在植物先天免疫过程发挥重要作用,但并不影响壳寡糖的诱导抗性。     

Effects of host plant environment and Ustilago maydis infection on the fungal endophyte community of maize (Zea mays)

The focus of many fungal endophyte studies has been how plants benefit from endophyte infection. Few studies have investigated the role of the host plant as an environment in shaping endophyte community diversity and composition. The effects that different attributes of the host plant, that is, host genetic variation, host variation in resistance to the fungal pathogen Ustilago maydis and U. maydis infection, have on the fungal endophyte communities in maize (Zea mays) was examined. The internal transcribed spacer (ITS) region of the rDNA was sequenced to identify fungi and the endophyte communities were compared in six maize lines that varied in their resistance to U. maydis. It was found that host genetic variation, as determined by maize line, had significant effects on species richness, while the interactions between line and U. maydis infection and line and field plot had significant effects on endophyte community composition. However, the effects of maize line were not dependent on whether lines were resistant or susceptible to U. maydis. Almost 3000 clones obtained from 58 plants were sequenced to characterize the maize endophyte community. These results suggest that the endophyte community is shaped by complex interactions and factors, such as inoculum pool and microclimate, may be important.     

Interactions between Fusarium verticillioides, Ustilago maydis, and Zea mays: an endophyte, a pathogen, and their shared plant host

Highly diverse communities of microbial symbionts occupy eukaryotic organisms, including plants. While many well-studied symbionts may be characterized as either parasites or as mutualists, the prevalent but cryptic endophytic fungi are less easily qualified because they do not cause observable symptoms of their presence within their host. Here, we investigate the interactions of an endophytic fungus, Fusarium verticillioides with a pathogen, Ustilago maydis, as they occur within maize (Zea mays). We used experimental inoculations to evaluate metabolic mechanisms by which these three organisms might interact. We assessed the impacts of fungal-fungal interactions on endophyte and pathogen growth within the plant, and on plant growth. We find that F. verticillioides modulates the growth of U. maydis and thus decreases the pathogen's aggressiveness toward the plant. With co-inoculation of the endophyte with the pathogen, plant growth is similar to that which would be gained without the pathogen present. However, the endophyte may also break down plant compounds that limit U. maydis growth, and obtains a growth benefit from the presence of the pathogen. Thus, an endophyte such as F. verticillioides may function as both a defensive mutualist and a parasite, and express nutritional modes that depend on ecological context.     

Infection of maize leaves with Ustilago maydis prevents establishment of C4 photosynthesis

The Basidiomycete fungus Ustilago maydis is the common agent of corn smut and is capable of inducing gall growth on infected tissue of the C4 plant maize (Zea mays). While U. maydis is very well characterized on the genetic level, the physiological changes in the host plant in response to U. maydis infection have not been studied in detail, yet. Therefore, we examined the influence of U. maydis infection on photosynthetic performance and carbon metabolism in maize leaf galls. At all stages of development, U. maydis-induced leaf galls exhibited carbon dioxide response curves, CO2 compensation points and enzymatic activities that are characteristic of C3 photosynthesis, demonstrating that the establishment of C4 metabolism is prevented in infected tissue. Hexose contents and hexose/sucrose ratio of leaf galls remained high at 6 days post infection, while a shift in free sugar metabolism was observed in the uninfected controls at that time point. Concomitantly, transitory starch production and sucrose accumulation during the light period remained low in leaf galls. Given that U. maydis is infectious on young developing tissue, the observed changes in carbohydrate metabolism suggest that the pathogen manipulates the developing leaf tissue to arrest sink-to-source transition in favor of maintaining sink metabolism in the host cells. Furthermore, evidence is presented that carbohydrate supply during the biotrophic phase of the pathogen is assured by a fungal invertase.     

Establishment of compatibility in the Ustilago maydis/maize pathosystem

The fungus Ustilago maydis is a biotrophic pathogen parasitizing on maize. The most prominent symptoms of the disease are large tumors in which fungal proliferation and spore differentiation occur. In this study, we have analyzed early and late tumor stages by confocal microscopy. We show that fungal differentiation occurs both within plant cells as well as in cavities where huge aggregates of fungal mycelium develop. U. maydis is poorly equipped with plant CWDEs and we demonstrate by array analysis that the respective genes follow distinct expression profiles at early and late stages of tumor development. For the set of three genes coding for pectinolytic enzymes, deletion mutants were generated by gene replacement. Neither single nor triple mutants were affected in pathogenic development. Based on our studies, we consider it unlikely that U. maydis feeds on carbohydrates derived from the digestion of plant cell wall material, but uses its set of plant CWDEs for softening the cell wall structure as a prerequisite for in planta growth.     

Reprogramming a maize plant: transcriptional and metabolic changes induced by the fungal biotroph Ustilago maydis

The fungal pathogen Ustilago maydis establishes a biotrophic relationship with its host plant maize (Zea mays). Hallmarks of the disease are large plant tumours in which fungal proliferation occurs. Previous studies suggested that classical defence pathways are not activated. Confocal microscopy, global expression profiling and metabolic profiling now shows that U. maydis is recognized early and triggers defence responses. Many of these early response genes are downregulated at later time points, whereas several genes associated with suppression of cell death are induced. The interplay between fungus and host involves changes in hormone signalling, induction of antioxidant and secondary metabolism, as well as the prevention of source leaf establishment. Our data provide novel insights into the complexity of a biotrophic interaction.     

Physical-chemical plant-derived signals induce differentiation in Ustilago maydis

Ustilago maydis is able to initiate pathogenic development after fusion of two haploid cells with different mating type. On the maize leaf surface, the resulting dikaryon switches to filamentous growth, differentiates appressoria and penetrates the host. Here, we report on the plant signals required for filament formation and appressorium development in U. maydis. In vitro , hydroxy-fatty acids stimulate filament formation via the induction of pheromone genes and this signal can be bypassed by genetically activating the downstream MAP kinase module. Hydrophobicity also induces filaments and these resemble the dikaryotic filaments formed on the plant surface. With the help of a marker gene that is specifically expressed in the tip cell of those hyphae that have formed an appressorium, hydrophobicity is shown to be essential for appressorium development in vitro . Hydroxy-fatty acids or a cutin monomer mixture isolated from maize leaves have a stimulatory role when a hydrophobic surface is provided. Our results suggest that the early phase of communication between U. maydis and its host plant is governed by two different stimuli.     

Use of PCR to detect infection of differentially susceptible maize cultivars using<Emphasis Type="Italic"> Ustilago maydis</Emphasis> strains of variable virulence

Ustilago maydis was specifically detected in infected maize plants by means of the polymerase chain reaction (PCR) using oligonucleotides corresponding to a specific region downstream of the homeodomain of the bE genes of the pathogen. The reaction gave rise to amplification of a ca. 500-bp product when tested with U. maydis DNA, but no amplification was detected with DNA from fungi not related to U. maydis. Using these primers, U. maydis was detected in infected maize plants from differentially susceptible cultivars as early as 4 days after inoculation with strains of variable degrees of virulence. Detection of U. maydis at early stages of infection, or in asymptomatic infected plants should assist in studies on plant-pathogen interactions.