利用微生物杀菌剂防治马铃薯黄萎病

马铃薯黄萎病是一种重要的世界性土传兼种传维管束病害,危害大且防治困难。利用活体微生物杀菌剂是防治作物土传病害的有效措施之一。本研究通过盆栽试验评价了微生物杀菌剂枯草芽胞杆菌可湿性粉剂对马铃薯出苗和生长的安全性,在河北省马铃薯主产区开展田间小区试验,研究了该制剂有效防治马铃薯黄萎病的使用方法和适宜施用剂量,并在河北省涞源县、围场县和永年区3县区分别开展了田间示范应用。盆栽试验和田间试验结果表明,枯草芽胞杆菌可湿性粉剂15、30和45 kg/hm²拌种处理对马铃薯出苗安全,对马铃薯生长没有不良影响;田间小区试验表明,在围场县试验田中,该制剂30 kg/hm²拌种处理单独使用或30 kg/hm²拌种加15 kg/hm²初花期滴灌使用均能显著减轻马铃薯黄萎病的发生,分别增产15.53%和17.10%;在新乐市试验田,枯草芽胞杆菌可湿性粉剂30 kg/hm²拌种处理显著增加马铃薯产量16.38%。田间示范应用结果表明,枯草芽胞杆菌可湿性粉剂30 kg/hm²拌种处理在涞源县和围场县防治马铃薯黄萎病效果显著,防效分别为84.22%和72.93%,两地分别显著增产24.30%和9.27%;在邯郸市永年区,相比化学药剂对照处理,枯草芽胞杆菌可湿性粉剂30 kg/hm²拌种处理显著增加马铃薯产量19.73%。本研究表明,枯草芽胞杆菌可湿性粉剂对马铃薯黄萎病具有显著的防治效果和显著的增产效果,为该制剂在马铃薯生产中高效应用提供了科学依据。     

贝莱斯芽胞杆菌次生抗生物质的研究进展

贝莱斯芽胞杆菌广泛分布于自然界,其次生抗生物质丰富,具有开发新型生物防治剂制剂的潜力。其基因组含有多种抗生物质的生物合成基因簇,如表面活性素、丰原素、杆菌霉素D等脂肽类化合物,杆菌烯、大环内酰亚胺、艰难菌素等聚酮类化合物,以及杆菌素和杆菌溶素等。除杆菌素以外,贝莱斯芽胞杆菌的绝大部分次生抗生物质依赖4'-磷酸泛酰转移酶（Sfp）途径合成。本文归纳和总结了贝莱斯芽胞杆菌抗生物质的化学结构、生物合成基因簇及其调控机制以及他们在控制植物、动物和人类疾病上的应用,以期为今后利用贝莱斯芽胞杆菌及其代谢产物研发生防制剂或临床药物提供科学依据。     

30亿CFU/g芽胞杆菌可湿性粉剂的研制及其对马铃薯黄萎病和疮痂病的防治效果

黄萎病和疮痂病是马铃薯生产上的重要土传病害,严重影响马铃薯的产量与品质,但缺乏有效的防治药剂。本研究以枯草芽胞杆菌HMB26553和解淀粉芽胞杆菌PHODG36为有效成分,通过对润湿剂、分散剂及紫外保护剂的筛选,确定了30亿CFU/g芽胞杆菌可湿性粉剂的配方(质量分数):HMB26553菌株母药10%、PHODG36菌株母药10%、润湿剂LT-569 1%、分散剂MF 2%、紫外保护剂抗坏血酸1%和滑石粉76%。该制剂芽胞数量37.6亿 CFU/g,杂菌率2.8%,pH 7.8,细度98.2%,干燥减量1.6%,润湿时间95.7 s,悬浮率80.7%。温室盆栽试验结果表明:该可湿性粉剂15~45 kg/hm2拌种处理对马铃薯出苗安全,对生长无不良影响,对马铃薯黄萎病的防治效果为45.1%~47.4%。田间小区试验结果表明:该可湿性粉剂15~45 kg/hm2拌种处理能显著减轻马铃薯疮痂病的发生,其中30 kg/hm2拌种处理防治效果高达72.0%,增产率达15.5%。不同地区的田间示范试验应用结果表明:该可湿性粉剂30 kg/hm2拌种处理对马铃薯黄萎病的防治效果为50.8%~62.1%,疮痂病为63.9%~65.7%,增产率为14.3%~29.4%。本研究结果表明,30亿CFU/g芽胞杆菌可湿性粉剂能有效防治马铃薯土传病害且增产作用明显,为该制剂在马铃薯生产上应用提供了科学依据。     

芽胞杆菌生物防治作用机理与应用研究进展

芽胞杆菌Bacillus spp.是国际上应用非常普遍的生防细菌，一直是当今土壤微生物学和微生态学研究热点之一。目前，国内外已完成多个芽胞杆菌菌株全基因组测序分析工作。本文从芽胞杆菌的次生代谢与抑菌作用、生物膜与竞争作用、植物-芽胞杆菌-病原菌互作与诱导植物抗性三方面概括性地综述了芽胞杆菌的生防机制。通过芽胞杆菌功能拓展的分子改良，可以提高植物抗病性，并就生防芽胞杆菌剂的商业化生产及在农业上的应用作了探讨。近年来我国学者在芽胞杆菌应用基础研究上取得了丰硕成果，为进一步实现芽胞杆菌的产业化及其应用奠定了坚实的基础。     

杀线虫解淀粉芽胞杆菌Sneb709与费氏中华根瘤菌Sneb183共培养发酵条件优化

根结线虫可引起作物发生严重的土传病害,生物防治是持续控制此类病害的有效措施.本实验室前期筛选获得多株杀线虫活性的芽胞杆菌和根瘤菌,本研究将解淀粉芽胞杆菌Sneb709与费氏中华根瘤菌Sneb183进行共培养后发现,共培养发酵液对南方根结线虫2龄幼虫J2活性显著提高,因此本文利用响应面法对其发酵条件进行优化,获得更好的生...     

微生物菌剂拌种对花生土传病害的防治效果及产量影响

为探索微生物菌剂拌种防治花生根部病害的防治效果及对产量的影响,以鲁花8号为试验材料,选择3个微生物菌制剂和1个化学药剂,设置1000亿CFU/mL解淀粉芽胞杆菌、200亿CFU/mL多粘类芽胞杆菌、1亿CFU/g真菌生防菌（TB）、1000亿CFU/mL解淀粉芽胞杆菌+1亿CFU/g真菌生防菌（TB）、200亿CFU/mL多粘类芽胞杆菌+1亿CFU/g真菌生防菌（TB）和25%咯菌腈拌种处理进行田间药效试验。结果表明： 6种供试药剂拌种处理均对花生有一定的增产作用,且对根腐病和白绢病均有一定的控制效果,其中1000亿CFU/mL解淀粉芽胞杆菌+1亿CFU/g真菌生防菌（TB）种子处理的出苗率、增产率和综合防效最高,其出苗率和增产率分别为94.35%和23.15%;出苗30 d后根腐病防效为76.95%,出苗60 d后根腐病和白绢病防效分别为83.54%和83.88%,在绿色防控技术推广应用中具有良好的应用前景。     

防治西葫芦和黄瓜白粉病的生物制剂的筛选

瓜类白粉病是蔬菜上的重要病害之一,为加大对该病害的生物防治力度,本研究采用课题组研发的1×109cfu/g玫瑰黄链霉菌水剂和10亿芽胞/g枯草芽胞杆菌悬浮剂以及市场上常用的枯草芽胞杆菌、哈茨木霉菌、寡雄腐霉、武夷菌素、多抗霉素等多种生物制剂进行田间药效试验,比较几种生物制剂的防治效果。结果表明,供试生物制剂对西葫芦白粉病的防效为57.65%~84.98%,其中3%多抗霉素水剂600倍液的防效最好,为84.98%,且具有明显的促生长作用,增产率达12.65%。其次为1 000亿芽胞/g枯草芽胞杆菌可湿性粉剂(武汉天惠)400倍液、2%武夷菌素水剂600倍液和3×108 cfu/g哈茨木霉菌可湿性粉剂300倍液,可作为西葫芦白粉病防治的选用药剂。1×109 cfu/g玫瑰黄链霉菌水剂对西葫芦和黄瓜白粉病均具有较好的防效,分别为78.68%和73.59%,具有开发和应用的市场价值。     

作物土传病害的危害及防治技术

近20年来,保护地在中国有了较大的发展,而保护地的发展和作物的连年栽培,导致土传病害和根结线虫发生越来越重,连续栽培3~5年后,作物产量和品质受到严重的影响,已成为生产中的突出问题。本文简述了我国重要作物如玉米、小麦、棉花、大豆、油菜的土传病害种类,以及高附加值作物黄瓜、番茄、茄子、辣椒、瓜类等作物的土传病害种类和20年来的变化。介绍了土传病害的防治方法,如农业防治包括轮作、抗性品种、嫁接、有机质补充、生物熏蒸、厌氧消毒;物理防治技术如太阳能消毒、蒸汽消毒、热水消毒、火焰消毒;化学防治技术如氯化苦、棉隆、威百亩、二甲基二硫、异硫氰酸烯丙酯、硫酰氟;生物防治技术如木霉、枯草芽胞杆菌、荧光假单胞菌、植物促生菌,以及预防为主的综合防治技术。种子、种苗消毒技术在本文中也进行了介绍。     

防治番茄灰霉病的枯草芽胞杆菌BAB-1粉尘剂研制

目前我国农药登记的微生物杀菌剂主要为水剂和可湿性粉剂两种剂型。为了克服常规喷雾导致温室大棚内湿度增加给防病带来的不利影响,本文以枯草芽胞杆菌BAB-1为有效成分开展了粉尘剂研究,通过对载体和助剂的筛选,明确了最佳的载体、助剂种类及其比例,最终确定100亿CFU/g枯草芽胞杆菌BAB-1粉尘剂的配方为：滑石粉20%、分散剂GY-D900 2%、菌株BAB-1原药补足至100%。该制剂具有很好的悬浮效果（30 min时悬浮率>50%）。室内盆栽试验表明,菌株BAB-1粉尘剂对番茄灰霉病防效达到57.14%~71.43%,对黄瓜灰霉病的保护作用优于治疗作用,施用1次的防病效果分别为48.80%和1.65%;温室大棚试验结果表明,该制剂在番茄大棚中施用3次后对灰霉病的防效达到79.04%,对照化学药剂嘧菌酯800倍水溶液的防效为82.55%。该研究结果为枯草芽胞杆菌BAB-1粉尘剂的进一步产业化开发和应用提供了科学依据。     

多粘类芽胞杆菌WY110对西瓜枯萎病的控制作用

西瓜枯萎病是由尖孢镰刀菌西瓜专化型引起的西瓜毁灭性土传病害,生产上主要采用药剂防治。由于病原菌的抗药性不断增强,利用拮抗菌对植物病害的生物防治越来越受到重视。目前,已有很多多粘类芽胞杆菌Paenibacillus polymyxa菌株用于植物病害的生物防治。     

Biological and application-oriented factors influencing plant disease suppression by biological control: a meta-analytical review

ABSTRACT Studies to evaluate the effectiveness of biological control in suppressing plant disease often report inconsistent results, highlighting the need to identify general factors that influence the success or failure of biological control in plant pathology. We conducted a quantitative synthesis of previously published research by applying meta-analysis to determine the overall effectiveness of biocontrol in relation to biological and application-oriented factors. For each of 149 entries (antagonist-disease combinations) from 53 reports published in Biological & Cultural Tests between 2000 and 2005, an effect size was calculated as the difference in disease intensity expressed in standard deviation units between the biocontrol treatment and its corresponding untreated control. Effect sizes ranged from -1.15 (i.e., disease strongly enhanced by application of the biocontrol agent) to 4.83 (strong disease suppression by the antagonist) with an overall weighted mean of 0.62, indicating moderate effectiveness on average. There were no significant (P >0.05) differences in effect sizes between entries from studies carried out in the greenhouse versus the field, between those involving soilborne versus aerial diseases, or among those carried out in conditions of low, medium, or high disease pressure (expressed relative to the disease intensity in the untreated control). However, effect sizes were greater on annual than on perennial crops, regardless of whether the analysis was carried out for all entries (P = 0.0268) or for those involving only soilborne diseases (P = 0.0343). Effect sizes were not significantly different for entries utilizing fungal versus bacterial biocontrol agents or for those targeting fungal versus bacterial pathogens. However, entries that used r-selected biological control agents (i.e., those having short generation times and producing large numbers of short-lived offspring) were more effective than those that applied antagonists that were not r-selected (P = 0.0312). Interestingly, effect sizes for entries that used Bacillus spp. as biological control agents were lower than for those that applied other antagonists (P = 0.0046 for all entries and P = 0.0114 for soilborne diseases). When only aerial diseases were considered, mean effect size was greater for entries that received one or two sprays than for those that received more than eight sprays of the biocontrol agent (P = 0.0002). This counterintuitive result may indicate that investigators often attempt unsuccessfully to compensate for anticipated poor performance in antagonist-disease combinations by making more applications.     

Improving biological control by combining biocontrol agents each with several mechanisms of disease suppression

ABSTRACT Two biocontrol agents, a yeast (Pichia guilermondii) and a bacterium (Bacillus mycoides), were tested separately and together for suppression of Botrytis cinerea on strawberry leaves and plants. Scanning electron microscopy revealed significant inhibition of Botrytis cinerea conidial germination in the presence of Pichia guilermondii, whereas Bacillus mycoides caused breakage and destruction of conidia. When both biocontrol agents were applied in a mixture, conidial destruction was more severe. The modes of action of each of the biocontrol agents were elucidated and the relative quantitative contribution of each mechanism to suppression of Botrytis cinerea was estimated using multiple regression with dummy variables. The improvement in control efficacy achieved by introducing one or more mechanisms at a time was calculated. Pichia guilermondii competed with Botrytis cinerea for glucose, sucrose, adenine, histidine, and folic acid. Viability of the yeast cells played a crucial role in suppression of Botrytis cinerea and they secreted an inhibitory compound that had an acropetal effect and was not volatile. Bacillus mycoides did not compete for any of the sugars, amino acids, or vitamins examined at a level that would affect Botrytis cinerea development. Viable cells and the compounds secreted by them contributed similarly to Botrytis cinerea suppression. The bacteria secreted volatile and non-volatile inhibitory compounds and activated the defense systems of the host. The nonvolatile compounds had both acropetal and basipetal effects. Mixture of Pichia guilermondii and Bacillus mycoides resulted in additive activity compared with their separate application. The combined activity was due to the summation of biocontrol mechanisms of both agents. This work provides a theoretical explanation for our previous findings of reduced disease control variability with a mixture of Pichia guilermondii and Bacillus mycoides.     

Evaluation of Trichoderma spp. as a Biocontrol Agent Against Soilborne Fungi and Plant-parasitic Nematodes in Israel

The optimal conditions required to market Trichoderma as a biocontrol agent against soilborne fungi and nematodes are discussed. These include a proper formulation, an efficient delivery system, and alternative methods for Trichoderma's application.The implementation of Trichoderma in integrated pest management (IPM) can be achieved using a soil treatment which combines reduced amounts of biocides/fungicides and the Trichoderma preparation. Biocontrol activity can be increased by combining two (or more) types of biocontrol agents. Moreover, the construction of a genetically modified Trichoderma can lead to the improvement of certain traits which are absent or not highly expressed in the native microorganism isolated from its natural habitat.Different Trichoderma harzianum and T. lignorum isolates were tested for their nematicidal activity against the root-knot nematode Meloidogyne javanica. In short-term experiments, improved growth of nematode-infected plants and decreases in the root-galling index and the number of eggs per gram of root were achieved when nematode-infested soils were pre- exposed to the T. harzianum preparations. A long-term experiment resulted in improved growth and higher yield of nematode-infected plants, but no significant change in the galling index, either by pre-exposure of the fungus to the soil or by enrichment in the root-ball.As biocontrol is an integral part of the IPM philosophy, judicious use of Trichoderma against soilborne pathogens, when demonstrated to be consistently effective, practical and economic, can serve as a model for the introduction and implementation of other biocontrol means into IPM.     

Biocontrol of soil-borne plant pathogens: Concepts and their application

A few soil-borne plant pathogens have been controlled successfully by commercial formulations of biocontrol agents, but many attempts to develop biocontrol inoculants, although promising under experimental conditions, have met with difficulties in practice. The reasons for this are discussed in this review, which outlines some of the major findings on the behaviour of microbial inoculants in soil. It is emphasized that biocontrol also occurs naturally in current agricultural practice and can be exploited purposely, but it is vulnerable to disruption by agrochemicals or mismanagement. The future of biocontrol of soilborne plant pathogens probably lies in integrated (biorational) control systems that combine the use of commercial inoculants, where appropriate, with management practices that maintain and enhance the natural biocontrol systems.     

植物病害生防芽孢杆菌抗菌机制与遗传改良研究

 芽孢杆菌是土壤和植物微生态的优势微生物种群,具有很强的抗逆能力和抗菌防病作用,许多性状优良的天然分离株已成功地应用于植物病害生物防治。芽孢杆菌抗菌防病机制包括竞争作用、拮抗作用和诱导植物抗病性。其中,核糖体合成的细菌素、几丁质酶和葡聚糖酶等抗菌蛋白以及次生代谢产生的抗生素与挥发性抗菌物质产生的拮抗作用是生防细菌最主要的抗菌机制。通过现代生物技术提高抗菌基因的表达水平和实现外源杀虫或抗菌基因的高效稳定共表达是增强生防芽孢杆菌抗菌活性和扩大防治对象的重要途径。基因组和蛋白组研究的迅猛发展必将极大地促进芽孢杆菌抗菌分子机制和抗菌基因工程研究的深入发展和广泛应用。     

Soilborne viruses: advances in virus movement,virus induced gene silencing,and engineered resistance

Until recently soilborne plant viruses were considered important only because they are causative agents for agricultural diseases. In recent years, soilborne plant viruses have played a significant role in advancing research into mechanisms of plasmodesmata transport, gene silencing, and engineered resistance to plant pathogens. Three different mechanisms by which viruses move through plasmodesmata have been identified using dianthoviruses, nepoviruses, and benyviruses. The infectious clone of the tobravirus Tobacco rattle virus (TRV) has become an important tool for studying virus induced gene silencing and may be a tool to silence meristematic gene expression. Investigations of soilborne viruses may enable the development of new strategies for control of soilborne diseases. The potential use of pathogen-derived resistance to control soilborne viruses is currently being explored in several laboratories.     

植物内生特基拉芽胞杆菌的分离、鉴定及防治西瓜枯萎病效果

为了筛选新的生防菌菌种资源,采用稀释涂布平板法从番茄植株中分离得到一株内生细菌wm031,采用平板对峙法研究其对病原菌的抑制效果,结合菌体特征、16S rDNA序列分析及生理生化指标,对其进行初步鉴定,明确分类地位,并采用温室栽培试验对其进行抗病效果初探。结果表明,该菌株对西瓜枯萎病菌具有显著拮抗作用,同时对油菜菌核病菌、小麦根腐病菌、小麦全蚀病菌、辣椒疫霉病菌、大蒜菌核病菌、水稻恶苗病菌、葡萄炭疽病菌和番茄枯萎病菌均具有广谱抗性。根据菌体特征、16S rDNA序列分析及生理生化指标的结果,初步鉴定该菌为特基拉芽胞杆菌Bacillus tequilens。该菌株能够在西瓜和水稻植株中稳定定殖。温室栽培试验表明,菌株wm031对西瓜枯萎病具有显著的防治效果74.6%,高出百菌清的防治效果14.1%,说明该菌株具有田间应用开发的潜力。     

Biological control of Ralstonia wilt,Phytophthora blight,Meloidogyne root-knot on bell pepper by the combination of Bacillus subtilis AR12, Bacillus subtilis SM21 and Chryseobacterium sp. R89

In a previous study, we screened the combination of the three bacterial strains—Bacillus subtilis AR12, Bacillus subtilis SM21, and Chryseobacterium sp. R89—(BBC), which can control mixed diseases, including Ralstonia wilt, Phytophthora blight, and Meloidogyne root-knot on bell pepper with high biocontrol efficacy and yield increase under greenhouse and field conditions. In this study, to achieve the best biocontrol agents for separately controlling Ralstonia wilt, Phytophthora blight and Meloidogyne root-knot by BBC, the biocontrol efficacy by BBC-related BCAs (AR12, SM21, R89, AR12 + SM21, AR12 + R89, SM21 + R89 and AR12 + SM21 + R89) were compared regarding all three single diseases and the mixed diseases, respectively. BBC achieved the highest biocontrol efficacy against all three single diseases and the mixed diseases (biocontrol efficacies of 92.46 %, 81.81 %, 86.49 % and 87.31 % against Ralstonia wilt, Phytophthora blight, Meloidogyne root-knot and the mixed diseases, respectively), which were significantly more than that attained by the three single strains and the two-strain combinations AR12 + SM21, AR12 + R89 and SM21 + R89. Although a biomass increase of 4.86–21.14 % was attained by BBC-related BCAs without BBC, BBC achieved a maximum biomass increase of 64.67 %.     

生物源杀菌剂与化学药剂协调防控番茄病害

为了阐明化学药剂与生防菌剂及植物源杀菌剂交替使用对番茄病害发展的影响,田间试验检测了交替施药的防病效果及病害发展曲线下面积.结果表明,枯草芽孢杆菌BAB-1水剂及化学药剂交替喷施7次对番茄灰霉病、叶霉病、晚疫病和早疫病的防效分别为86.9%,61.0%,72.3%和77.2%.植物源杀菌剂1-氧-乙酰基大花旋覆花内酯(ABL)乳油与化学药剂及生防菌剂枯草芽孢杆菌BAB-1水剂交替喷施9次对上述病害的防效分别为88.6%,87.4%,85.4%和83.2%,相当于不同作用机制的化学药剂交替喷施9次的防效.此外,化学药剂与BAB-1水剂及ABL乳油交替喷施明显延缓了这4种病害的发展,减少了化学药剂施用次数.     

Pseudomonas biocontrol agents of soilborne pathogens: looking back over 30 years

ABSTRACT Pseudomonas spp. are ubiquitous bacteria in agricultural soils and have many traits that make them well suited as biocontrol agents of soilborne pathogens. Tremendous progress has been made in characterizing the process of root colonization by pseudomonads, the biotic and abiotic factors affecting colonization, bacterial traits and genes contributing to rhizosphere competence, and the mechanisms of pathogen suppression. This review looks back over the last 30 years of Pseudomonas biocontrol research and highlights key studies, strains, and findings that have had significant impact on shaping our current understanding of biological control by bacteria and the direction of future research.