水稻白叶枯病菌在离体培养条件下生物膜形成的检测

分析了不同培养和测试条件(塑料和玻璃表面、培养时间、培养温度和培养基)对水稻白叶枯病菌(Xanthomonas oryzae pv. oryzae,Xoo)生物膜形成的影响,并测定了不同地区来源的Xoo野生型菌株、基因缺失突变体的生物膜形成。结果表明,在聚苯乙烯表面生长、用M210培养基在23 ℃下静止培养24 h是Xoo生物膜形成比较适宜的条件;不同地区来源的菌株生物膜形成能力不同;鞭毛相关基因fleQxoo、fliExoo和rbfCxoo以及H2O2降解调控基因oxyRxoo的缺失突变均影响生物膜形成。     

《植物保护》2011年第1期各栏目最受读者关注的文章

专论与综述1.傅本重.植物与病原物互作中的生物膜研究2.罗沙,余柳青,刘都才,等.稻田稗草对二氯喹啉酸的抗药性研究进展研究报告     

群体感应调控因子ComA对枯草芽胞杆菌NCD-2抑菌物质产生和生物膜形成的影响

为明确群体感应系统在枯草芽胞杆菌NCD-2菌株抑菌活性和生物膜形成中的作用,通过同源重组技术对群体感应系统中的comA基因进行定位缺失突变,分别比较了NCD-2菌株和comA基因突变子在胞外酶合成、抑菌活性、脂肽抗生素(丰产素)产生以及生物膜形成上的差异,并进一步利用实时荧光定量RT-PCR技术比较了二者生物膜基质编码基因epsA和tasA的表达情况。结果表明,通过同源重组技术获得了comA基因突变子MA-4,同菌株NCD-2相比,其在胞外蛋白酶和纤维素酶的合成能力、对番茄灰霉菌Botrytis cinerea的抑菌活性、丰产素的合成量和生物膜的形成能力上均明显下降;生物膜基质编码基因tasA的表达量下降了70%,而epsA的表达量变化不明显。表明ComA是NCD-2菌株脂肽抗生素和生物膜形成中的重要调控因子。     

ywbB基因对枯草芽胞杆菌NCD-2菌株生物膜形成和根际定殖能力的影响

为明确枯草芽胞杆菌Bacillus subtilis NCD-2菌株生物膜形成相关基因及其在生物膜形成、根际定殖中的作用,采用转座子插入技术,从4000余株NCD-2突变子中获得6株生物膜形成能力降低的突变子。在生物膜形成能力丧失的突变子M3中,转座子以单拷贝的形式插入到ywbB基因内部。通过基因同源重组技术对ywbB基因进行缺失突变,获得ywbB基因缺失突变菌株NCD-2(ΔywbB),与野生菌株相比,突变子丧失了生物膜形成能力。进一步比较发现,突变菌株NCD-2(ΔywbB)显著降低了在棉花根际的群体数量,同时也降低了对棉花立枯病的防治效果。研究证明,ywbB是NCD-2菌株生物膜形成途径中的重要基因,且生物膜形成与NCD-2菌株根际定殖和生防效果呈正相关。     

L-脯氨酸对枯草芽胞杆菌NCD-2菌株生物膜形成的影响

 枯草芽胞杆菌 NCD-2菌株在棉花品种‘冀棉11'根际的定殖能力与其根系分泌物中的L-脯氨酸相关。本研究通过外源添加L-脯氨酸评价其对NCD-2菌株生长量和生物膜形成的影响。结果表明,在MSgg培养基中添加L-脯氨酸培养48 h后对菌株生长不存在显著影响。采用称重法测定生物膜的湿重和干重,结果表明在MSgg培养基添加L-脯氨酸显著增加了生物膜产量,且生物膜的褶皱程度和立体结构变得更为明显。采用结晶紫染色法进行生物膜的定量测定发现,浓度为1.250～10.000 mg·mL^-1的L-脯氨酸可以显著提高菌株生物膜形成能力,而低浓度0.625 mg·mL^-1 的L-脯氨酸对生物膜形成无影响。通过RT-qPCR检测了NCD-2菌株生物膜形成相关基因tasA和epsA表达量的变化,结果表明,添加上述不同浓度L-脯氨酸能够显著提高tasA基因表达量,提高了2.53~9.98倍。除了浓度为0.625 mg·mL^-1 L-脯氨酸可提高epsA基因表达外,其他浓度没有显著改变epsA基因的表达,而且epsA基因表达量低于tasA基因。综合分析表明,L-脯氨酸促进NCD-2菌株生物膜形成不是通过改变菌株生长量,而是诱导tasA基因的表达产生更多的胞外蛋白TasA。     

棉花根系分泌物对枯草芽胞杆菌NCD-2生物膜形成和根际定殖的影响

 为明确棉花根系分泌物对生防枯草芽胞杆菌NCD-2生物膜形成的影响,本研究首先比较了菌株NCD-2在不同棉花品种(冀棉11、中棉所41、中植棉2号、鲁棉29和海岛棉Pima90)根际的定殖能力。结果表明,菌株NCD-2在Pima90的根际定殖能力最强,播种35 d后在根际的群体数量达到7.63×10⁵ CFU·g^-1根重;而在中植棉2号的根际定殖能力最弱,播种35 d后根际菌株NCD-2的群体数量为6.51×10⁴ CFU·g^-1根重。通过再循环水培系统收集根系分泌物的方法获得了不同棉花品种的根系分泌物,测定了其对菌株NCD-2生物膜形成的影响。结果表明,海岛棉Pima90的根系分泌物对该菌株生物膜形成的促进作用最强,而中植棉2号的根系分泌物对菌株NCD-2生物膜形成影响最弱。进一步分析了棉花根系分泌物中6种糖分和13种氨基酸对菌株NCD-2生物膜形成的影响,结果发现,葡聚糖和脯氨酸显著促进了该菌株的生物膜形成。     

表面活性素促进枯草芽胞杆菌XF-1在大白菜叶际定殖能力研究

枯草芽胞杆菌XF-1是一株大白菜内生生防菌,通过叶面喷施的方式可有效防控大白菜根肿病,但其叶际定殖的分子机制尚不明确。为了明确表面活性素对XF-1在大白菜叶际定殖的影响,进一步提高其在田间应用效果,本研究通过测定生长曲线、泳动运动、群集运动和24孔细胞培养板静置培养试验,分析表面活性素对XF-1菌株生长速率、运动能力和生物膜形成水平的影响;利用叶片-微生物互作分析和定殖试验,测定XF-1及突变体XF-1-ΔsrfA在大白菜叶表黏附和植株内的定殖能力。结果表明,XF-1-ΔsrfA与XF-1相比生长曲线没有明显差异,泳动能力和群集运动能力分别显著下降了36.8%和43.9%,静置培养24 h后生物膜形成能力显著下降53.9%,48 h后无显著差异。叶片-微生物互作试验中,突变体标记菌XF-1-ΔsrfA-gfp黏附效率较XF-1-gfp约下降80%。喷施接种3~7 d后, XF-1-ΔsrfA-gfp相较于XF-1-gfp在大白菜叶片内和根系中的定殖数量均显著下降。试验结果表明表面活性素对枯草芽胞杆菌XF-1在大白菜叶际定殖具有较为明显的促进作用,srfA基因的缺失显著抑制XF-1的叶际定殖能力。     

臭氧-生物活性炭处理西北村镇微污染窖水实验研究

针对我国西北村镇分散式水窖微污染窖水的水质特性,探讨臭氧-生物活性炭工艺(03-BAC)处理微污染窖水的可行性.结果表明,活性炭挂膜过程可分为三个阶段:微生物适应期、生物膜生长期、生物膜成熟期.煤质活性炭在生物膜成熟期对窖水中CODMn、氨氮的去除率分别为81.02％、77.21％,适合作为生物活性炭工艺(BAC)的填料.预臭氧最佳投加量为3mg/L,03-BAC出水各项指标均符合生活饮用水卫生标准.03-BAC工艺适合处理西北村镇分散式水窖微污染窖水.     

水稻白叶枯病菌HD-GYP结构域蛋白PXO_03945的功能鉴定

为了揭示水稻白叶枯病菌(Xanthomonas oryzae pv.oryzae,简称Xoo)环二鸟苷酸(c-di-GMP)信号相关蛋白PXO_03945的生物学功能,本研究通过基因同源重组和无标记交换,对PXO_03945基因进行了缺失突变,对野生型、突变体和互补菌株进行表型测定,分析该基因缺失突变对病菌胞内c-di-GMP水平、致病性、运动性、胞外多糖产生和生物膜形成的影响。结果表明,PXO_03945蛋白具有参与c-di-GMP降解的磷酸二酯酶(PDE)的HD-GYP结构域和功能未知的DUF3391结构域。全长基因缺失可导致体内c-di-GMP浓度明显上升。与野生型相比,ΔPXO_03945对水稻品种日本晴的致病性显著下降,而游动性增强,胞外多糖产生和生物膜形成增加。基因互补可以使之恢复。因此,HD-GYP结构域蛋白PXO_03945可能通过降解胞内c-di-GMP,正向调控了致病性,负向调控了运动性、EPS产生和生物膜形成能力。     

西瓜嗜酸菌LuxR家族基因luxR_(4229)的功能研究

瓜类细菌性果斑病是葫芦科作物上的一种严重的世界性病害,其病原菌为西瓜嗜酸菌(Acidovorax citrulli)。本研究以野生型菌株Ac-5为研究对象,通过同源重组双交换的方法,构建了LuxR家族功能基因luxR_(4229)的全基因缺失突变株,并对突变株进行致病性、生物膜形成能力和运动能力等生物学特性测定。同时,qRT-PCR定量检测了鞭毛基因fliR和fliC以及III型分泌系统功能基因hrpE和hrcN的表达水平。结果显示:与野生型相比,突变株致病力明显降低,运动能力减弱,而生物膜形成能力增强。fliR、fliC、hrpE和hrcN基因在突变株中的表达量下调,luxR4229对fliR、fliC、hrpE和hrcN基因均为正调控。基因luxR_(4229)在对西瓜嗜酸菌致病力、运动能力及生物膜的形成具有重要调控作用。     

Products containing microorganisms as a tool in integrated pest management and the rules of their market placement in the European Union

Products containing microorganisms (bacteria, fungi and viruses) can be used in plant production as an intervention as well as a prevention method for pest control. Their utilisation is strictly in line with the principles of integrated pest management, provided that they are effective and safe. The rules of registration of microorganisms for crop production in the European Union differ, depending on whether they are placed on the market as plant protection products or not. For over 20 years, uniform rules for registration of plant protection products have been in force. Currently, 36 microorganisms marked up to the strain are approved for use in pest control in the Community. The decision concerning market placement of plant protection products containing approved microorganisms is issued for each member state separately. The approaches to market placement of other products with microorganisms differ within the EU, ranging from a complete lack of requirements to long and costly registration procedures. © 2015 Society of Chemical Industry     

叶圈微生物的群落结构与生态功能研究进展

植物叶圈和根际是两个独特、复杂的生态系统,含有丰富的微生物种类,在促进植物生长,增强植物抗病性、抗逆性等方面有着非常重要的作用。在近几十年内的研究中,根际一直是微生物学家和植物学家研究和关注的焦点,特别是在根际微生物促进植物生长和增强植物抗病性的效果和机理方面取得了重要的研究进展。相对而言,对于叶圈微生物的研究远远滞后于对根际微生物的研究。叶圈微生物最初主要是对植物致病微生物进行研究,但是由于技术条件(主要是采用传统可培养方法进行研究)的限制,叶圈微生物研究进展缓慢。2000年以后,随分子生物学技术的发展,特别是高通量测序技术的发展,叶圈微生物研究进入了快速发展段。研究发现,叶圈微生物中只有少数致病,而大多数能够为植物提供养分、促进植物生长、提高植物抗病性和抗逆性,在植物群落组成和生态功能方面发挥了重要作用。通过介绍叶圈定义的提出及发展过程,阐述了叶圈微生物在研究方法、生物学功能及作用机理方面的研究进展,并对叶圈微生物未来发展方向进行了展望。     

昆虫共生微生物在病虫害防治的研究进展

昆虫体内的共生微生物是一大类生物群体，包括细菌、真菌、病毒及一些小型原生生物，它们广泛分布于昆虫体内，具有参与昆虫体内合成氨基酸、维生素等小分子营养物质，分解纤维素等大分子化合物，以及降解杀虫剂和植物次生代谢物等作用。此外，也能间接调控宿主生物的免疫系统，从而阻断病原体的复制和传播。因此，在害虫防治和控制虫媒病毒传播等方面具有应用潜力。该文综述昆虫共生微生物在病虫害防治方面的最新研究进展，探讨其生物学功能、与宿主的作用机制、宿主对病原微生物的适应机制以及在病虫害防治中应用等方面的研究进展，并对未来发展趋势及在害虫防治方面的应用前景进行展望。     

Contribution of the seed microbiome to weed management

Seed‐attacking microorganisms have an undefined potential for management of the weed seedbank, either directly through inundative inoculation of soils with effective pathogenic strains, or indirectly by managing soils in a manner that promotes native seed‐decaying microorganisms. However, research in this area is still limited and not consistently successful because of technological limitations in identifying the pathogens involved and their efficacy. We suggest that these limitations can now be overcome through application of new molecular techniques to identify the microorganisms interacting with weed seeds and to decipher their functionality. However, an interdisciplinary weed management approach that includes weed scientists, microbiologists, soil ecologists and molecular biologists is required to provide new insights into physical and chemical interactions between different seed species and microorganisms. Such insight is a prerequisite to identify the best candidate organisms to consider for seedbank management and to find ways to increase weed seed suppressive soil communities.     

Authorization of plant protection products containing microorganisms in the European Union1

The main principles concerning the authorization of plant protection products containing microorganisms in the European Union are explained: a positive list of microorganisms to be adopted by the Commission, authorization of individual plant protection products by the Member States according to common criteria and data requirements, obligation of mutual recognition of authorizations granted by other Member States subject to comparable conditions. Further detailed provisions are under preparation for the common criteria and data requirements as well as for the rationalization of the procedures for plant protection products containing genetically modified microorganisms.     

烟嘧磺隆的微生物降解研究进展

烟嘧磺隆属磺酰脲类除草剂,因其对玉米安全,对一年生和多年生禾本科杂草及部分阔叶杂草、莎草科杂草高效而在玉米田广泛应用,但其在土壤中的残留则易对后茬敏感作物产生药害。微生物对烟嘧磺隆的降解有望成为修复污染土壤的有效措施。文章从烟嘧磺隆的使用及危害,可降解烟嘧磺隆的微生物种类及降解特性、降解途径、降解酶及其编码基因等方面进行了综述。目前有关烟嘧磺隆微生物降解的研究多集中于其降解菌的筛选、降解特性及降解途径等方面,对于其微生物降解过程中起关键作用的酶和基因方面的研究仍较少,因此未来的研究趋势将主要体现在降解复合菌系的培养、降解微生物的环境生态学、降解基因以及降解酶制剂等方面。     

黄葵茎腐病病原菌的分离与鉴定

针对江苏宜兴黄葵主产区发生的黄葵茎腐病,通过分离黄葵植株不同部位(根、茎、叶)以及根际土壤、根围土壤的微生物,经形态学观察,结合分子进化分析构建了黄葵不同生态位的微生物物种系统发育树,对其种类和分布进行了分析。结果表明,细菌优势类群为芽胞杆菌,而真菌无明显优势类群。同时,根据柯赫氏法则测定了分离得到的微生物菌株的致病性,确定了菌株F28可引起黄葵茎腐病,经ITS分子进化分析及形态学观察,将其鉴定为变紫青霉Penicillium sanguineum。     

Phytotoxicants from microorganisms and related compounds

Phytotoxic compounds produced by microorganisms are reviewed. Their utilisation as leads to new herbicides is explored in three chemical classes : 2-aminoalk-3-enoic acids, ether derivatives of 3-hydroxycyclobut-3-ene-1,2-diones, and 2-(acylaminooxy)acetic acid derivatives. It is concluded that some bacteria and fungi yield compounds possessing sufficient herbicidal activity to be valuable as herbicides as such, or as leads for chemical optimisation.     

Selbst oder Nicht-Selbst �C die Rezeptoren des pflanzlichen Immunsystems

Plants recognize conserved molecular structures from microorganisms, which triggers active immune responses. Successful pathogens have to overcome this level of immunity; however, plants in turn can adapt their immune system, thus plants and pathogens are in an evolutionary arms race. As being sessile organisms, plants need to integrate and adapt to changing environmental conditions such as light, temperature, drought, or microorganisms. Plants protect themselves against diseases through sensitive recognition of potential pathogens and effective defense systems. The first level of the plant immune system provides recognition of a broad spectrum of microorganisms leading to defense activation (Bittel and Robatzek 2007). The second level of the plant immunity allows certain plant cultivars to detect of specific pathogen strains??a phenomenon also referred to as ??gene-for-gene resistance?? (Jones and Dangl 2006). The first level of immunity occurs rapidly and triggers active defenses normally without harm to the plant cell. The second level of plant immunity develops over days and deploys a local cell death, which prevents pathogens from further spread into tissues. In addition to these cell-autonomous defense systems, plants have also evolved strategies of systemic immunity.     

Control of seed-borne pathogens on legumes by microbial and other alternative seed treatments

Greenhouse trials were carried out in order to test the efficacy of different seed treatments as alternatives to chemicals against Colletotrichum lindemuthianum cause of anthracnose on bean and Ascochyta spp. cause of Ascochyta blights on pea, respectively. Resistance inducers, commercially formulated microorganisms, non-formulated selected strains of different microorganisms (fungi, bacteria and yeasts) and plant extracts were applied as dry or liquid seed treatments on naturally infested seeds. Seedling emergence and disease incidence and/or severity were recorded. Almost all seed treatments turned out to be ineffective in controlling the Ascochyta infections, which is in line with the literature stating that these pathogens are difficult to control. The only alternative treatments that gave some control of Ascochyta spp. were thyme oil and a strain of Clonostachys rosea. The resistance inducers tested successfully controlled infections of bean by C. lindemuthianum. Among the formulated microorganisms, Bacillus subtilis-based formulations provided the best protection from anthracnose. Some strains of Pseudomonas putida, a disease-suppressive, saprophytic strain of Fusarium oxysporum and the mustard powder-based product Tillecur also proved to be effective against bean anthracnose. However, among the resistance inducers as well as among the other groups, certain agents caused a significant reduction of plant emergence. Different alternative seed treatments can therefore be used for the control of C. lindemuthianum on bean, while on pea only thyme oil and a strain of Clonostachys rosea showed some effectiveness against Ascochyta spp.