健康与罹患青枯病的番茄土壤细菌群落特征比较

应用实时荧光定量PCR及MiSeq高通量测序技术,全面地研究了连作番茄田块中健康与感染青枯病植株周围土体及根际土壤细菌群落结构和组成。结果表明:健康番茄土体土壤的pH及全碳含量显著高于感病番茄土体土壤;土体及根际土壤的细菌群落结构和组成明显不同于感病番茄土体及根际土壤细菌群落。与感病番茄根际相比,健康番茄根际细菌的数量显著升高而青枯菌数量显著降低;细菌群落的Shannon多样性指数显著增高;拟杆菌门及其所含的噬几丁质菌属、金杆菌属、动杆菌属、黄杆菌属及Taibaiella的相对丰度显著增高而变形菌门及其所含的青枯菌属的相对丰度显著降低。综上,抑制土传青枯病发生的番茄根际土壤细菌群落特征明显,其生物量及多样性高,土著有益菌群数量多而病原菌数量少,为番茄土传青枯病的生物防控提供了指导方向与理论依据。     

根际促生菌防控土传病害的机理与应用进展

根际促生菌作为一类活跃于植物根际的有益微生物,对抑制土传病害的发生,改善和维护土壤生态质量具有重要作用以及广阔的发展前景。本文综述了根际促生菌直接或间接的土传病害防控机理,包括产抗生素、产水解酶、释放挥发性气体、诱导抗性、分泌铁载体、分泌激素和固氮解磷等;亦从信号识别与迁移、定殖规律以及定殖的微生物群落影响等方面对根际促生菌的定殖防病机理予以了总结;此外,笔者还就根际促生菌的应用现状进行了概述。     

生物有机肥对烟草根际微生物群落及青枯雷尔氏菌丰度的影响

【目的】探究生物有机肥施用对烟草根际土壤细菌、真菌群落结构和青枯雷尔氏菌丰度的影响机理。【方法】选用长沙市某公司生产的生物有机肥和常规烟草专用肥,在湘西花垣县长期定位试验点连续5年开展大田试验,研究施肥对土壤理化性质和微生物群落的影响。试验设置两种施肥处理：常规烟草专用肥（CF）和生物有机肥（BOF）。【结果】与CF相比,施用生物有机肥处理的土壤烟草青枯病发病率降低了89.8%,同时青枯雷尔氏菌相对丰度也显著降低,降幅达40.1%;土壤pH、碱解氮和有效磷显著增加,分别增加了1.2%、12.1%和60.2%;施用生物有机肥后根际土壤微生物如Roseiflexaceae,Gemmatimonadaceae,Nitrospira,Ramophialophora,Preussia等显著富集,且这些潜在有益菌与青枯雷尔氏菌相对丰度呈显著负相关关系。通过ABT预测模型分析发现潜在有益菌是影响青枯雷尔氏菌相对丰度的最主要生物因子。【结论】连续5年的试验结果表明,施用生物有机肥不仅改善了作物生长的土壤环境,显著提高了土壤pH和土壤速效养分含量,还促使潜在有益菌在根际土壤中富集,抑制了青枯雷尔氏菌的生...     

生物质炭介导生防微生物抑制辣椒疫霉的作用

生物质炭可有效防控土传病害,筛选并鉴定出生物质炭介导下的生防微生物,可为研究生物质炭防病机理和强化生物质炭防病效果提供理论依据。本研究首先进行秸秆生物质炭防控辣椒疫病盆栽试验,利用定量PCR和平板计数明确生物质炭在防控辣椒疫病时可富集的已知生防微生物,再通过选择性培养基初筛和定殖复筛筛选出生物质炭可富集的潜在生防微生物菌株,最后研究各菌株在土壤中对辣椒疫霉的抑制作用。结果表明,秸秆生物质炭使根际辣椒疫霉数量显著降低95.1%、辣椒疫病发生率显著降低91.1%,并使具有生防功能的木霉菌、青霉菌、曲霉菌、芽孢杆菌、假单胞菌和鞘氨醇单胞菌数量显著增加2.22倍、4.09倍、3.89倍、2.45倍、1.45倍和1.30倍。通过平板初筛得到可能被生物质炭富集的22株潜在生防菌株。定殖复筛剔除部分假性生物质炭介导菌株,获得可明确被生物质炭富集的2株木霉菌、3株青霉菌、2株曲霉菌、3株芽孢杆菌、3株假单胞菌、3株链霉菌和2株鞘氨醇单胞菌。木霉菌（TR1和TR3）、青霉菌（PE1）、曲霉菌（AS1和AS2）、芽孢杆菌（BA1、BA2和BA3）、假单胞菌（PS1和PS3）、链霉菌（ST1、ST4和ST5）13个菌株可显著削减土壤辣椒疫霉数量。其中,所有木霉菌和曲霉菌菌株（TR1、TR3、AS1和AS2）及芽孢杆菌（BA1和BA2）、假单胞菌（PS1和PS3）和链霉菌（ST1）9个菌株与生物质炭具有显著的协同抑制辣椒疫霉效果。因此,防控辣椒疫病时,木霉菌、曲霉菌、芽孢杆菌、假单胞菌和链霉菌是生物质炭介导下的主要防病微生物。     

不同基质生物有机肥防控番茄土传青枯病及促生效果研究

本文旨在研究功能菌株解淀粉芽孢杆菌配合不同原料的有机肥，制备不同生物有机肥对番茄防控土传青枯病以及促生的效果研究，并考察不同生物有机肥对土壤养分及根际细菌群落的影响，获得能够有效防控番茄青枯病的生物有机肥。本研究以解淀粉芽孢杆菌T-5为功能菌株，分别配合秸秆有机肥及鸡粪有机肥，经二次发酵分别制得秸秆生物有机肥（BIO1）和鸡粪生物有机肥（BIO2）。于江苏省南京市一处青枯病多发的番茄大棚进行试验。考察不同施肥处理下番茄的发病情况、生长情况以及土壤理化性质，并利用高通量测序探究根际细菌群落的变化。两种生物有机肥均能显著降低番茄土传青枯病的发病率，且BIO1处理的防控效果最佳，使发病率降低了94.45%。两种生物有机肥均能显著促进番茄生长，增加了番茄地上部的生物量并提高了产量，但对于番茄果实品质并无显著提升。此外，施用BIO1可显著增加土壤有机质含量，BIO2则显著提高了土壤pH。土壤有机质与番茄的发病率呈负相关，与番茄地上部生物量和产量呈显著正相关关系。基于高通量测序结果，发现生物有机肥可显著提高根际细菌多样性，同时改变了细菌群落结构。防控效果最好的BIO1处理中，变形菌门相对丰度最高，放线菌门相对丰度最低，土壤硝态氮和pH与这两个门水平的细菌类群具有显著相关性。以解淀粉芽孢杆菌为功能菌株、秸秆为原料制备的秸秆生物有机肥可有效防控番茄土传青枯病，效果优于鸡粪生物有机肥。该生物有机肥还可显著促进番茄生长及产量提升，并通过调控根际细菌群落中变形菌门和放线菌门的相对丰度，提高土壤抑病能力，减少土传青枯病的发生。     

生物有机肥育苗防控烟草青枯病

【目的】烟草青枯病是影响贵州烟叶生产的主要土传病害之一。生物防控方法作为防控烟草青枯病体系的有效措施之一,寻求经济可行的途径势在必行。【方法】利用实验室自主筛选到的烟草青枯菌拮抗菌NJL-14,通过二次固体发酵研制成烟草专用拮抗青枯病型生物有机肥 (BIO),选取K326和红花大金元两个烟草品种分别进行育苗和盆栽试验。育苗床基质分别添加BIO 1%、2%、3%、4%和5%,以确定适宜的BIO育苗添加量。盆栽试验两个品种分别设定五个处理:常规基质育苗 + 植烟健康土壤(CKn+CKp）、常规基质育苗?+?青枯病病土（CKn+Rs）、常规基质育苗?+?青枯病病土?+?有机肥(CKn+OFp)、常规基质育苗?+?青枯病病土?+?BIO(CKn+BIOp)、育苗和盆栽病土中都施入BIO(BIOn+BIOp)。分别采用荧光定量PCR和Biolog-ECO研究育苗基质和盆栽土壤中细菌和真菌数量变化,以及微生物功能多样性,特别是烟草青枯菌及其拮抗菌的消长关系。【结果】当BIO浓度在2%以内时,烟草的发芽率与对照无显著差异;与普通漂浮育苗相比,托盘育苗不但能稳定和延长拮抗菌NJL-14在根际的定殖,而且显著提高烟苗生物量,K326经BIO育苗处理的烟苗地上部和地下部的干重分别比常规基质育苗的处理高17.2%和30.8%,而红花大金元分别高14.9%和20.0%;采用育苗与移栽土壤双重使用BIO模式对烟草青枯病的防控效果明显,防控效果可以达到100%,明显优于只在盆栽中使用BIO模式,并且能够显著抑制土壤中病原菌数量在107 cfu/g土以下,同时有效提高土壤微生物生态多样性。【结论】采用育苗与移栽土壤双重使用烟草专用拮抗青枯病型生物有机肥模式可有效防控烟草青枯病,控制土壤中病原菌数量,改善土壤微生物功能多样性,为有效地生物防控烟草青枯病奠定基础。     

病原青枯菌土壤存活的影响因素研究进展

土传青枯病是一种毁灭性的细菌性病害,广泛分布于热带、亚热带和温带地区,严重威胁世界粮食安全。病原青枯菌主要从土壤中侵染作物根系,其在土壤中存活能力强,因此防治极为困难。明确病原青枯菌土壤存活的关键影响因素有助于开发高效阻控土传青枯病的措施。国内外学者在青枯菌的土壤存活方面开展了大量研究,但由于影响青枯菌土壤存活的因素复杂,而相关研究多围绕单一因素展开,缺乏针对青枯菌土壤存活规律和影响因素的系统性认识。本文系统梳理了青枯菌的自身特性（基因、行为和代谢产物）及土壤生物、非生物因素对其在土壤中存活的影响,阐明了青枯菌在寄主存在时土体存活、向寄主根表方向运动迁移时根际存活以及入侵寄主根系时根表存活的主要影响因子,以期为土传青枯病的系统阻控提供参考。     

丛枝菌根真菌提高感染青枯菌番茄根际土壤细菌群落多样性和稳定性及有益菌属相对丰度

  【目的】  青枯病是由茄科雷尔氏菌 (Ralstonia solanacearum, 亦称青枯菌) 诱导产生的一种高温高湿型土传病害,土壤温度高、湿度大时易于青枯菌的繁殖进而引发青枯病。丛枝菌根真菌 (arbuscular mycorrhiza, AM) 可能通过调控根际微生物区系对病原体产生影响,我们研究了AM真菌对青枯菌入侵条件下土壤细菌群落的影响。  【方法】  以番茄 (Solanum lycopersicum) 为试材进行盆栽试验,供试AM真菌为摩西管柄囊霉 (Funneliformis mosseae) M47V,供试病原菌为茄科雷尔氏菌QL-RS 1115 (GenBank:GU390462)。催芽5日的番茄种子,接种AM菌剂的为菌根苗,未接种AM真菌的为非菌根苗。在番茄幼苗生长30天时,一半菌根苗和非菌根苗接种青枯菌,另一半不接种青枯菌,共4个处理。在接种青枯菌后1天和14天,采集番茄样品,采用抖土方法采集根际土壤,利用实时荧光PCR分析番茄根际青枯菌数量,采用16S rRNA高通量测序探究土壤细菌群落多样性和结构稳定性。  【结果】  在接种青枯菌初期 (1天),非菌根苗接种青枯菌 (TR–AMF) 和菌根苗接种青枯菌 (TR+AMF) 两组处理的根际土壤细菌群落结构发生明显改变,Chao1指数、Shannon指数和Simpson指数显著降低 (P<0.05),共现网络的节点数和连接数明显减少,模块化程度降低,共现网络简化表明细菌群落结构的稳定性降低。接种青枯菌14天后,不动杆菌属 (Acinetobacter)、鞘氨醇单胞菌属 (Sphingomonas)、溶杆菌属 (Lysobacter)、假单胞菌属 (Pseudomonas) 等有益细菌属在感染青枯菌的番茄根际富集,细菌共现网络的节点数和连接数增加,模块化程度提高,表明细菌群落稳定性得到恢复。与非菌根苗相比,菌根苗接种青枯菌 (TR+AMF) 和菌根苗未接种青枯菌 (TN+AMF) 两个处理番茄根际土壤中青枯菌丰度显著降低 (P<0.05)。AM真菌显著提高Chao1指数和Shannon指数 (P<0.05),提高了感染青枯菌番茄根际土壤中黄杆菌属(Flavobacterium)、黄色土源菌属 (Flavisolibacter)、噬胞菌属 (Cytophaga) 和苔藓杆菌属 (Bryobacter) 的相对丰度,同时增加了共现网络的节点数和连接数,并促进番茄根际细菌物种之间的良性互作,提高细菌网络的复杂程度。  【结论】  感染青枯菌的番茄根际会富集不动杆菌属 (Acinetobacter)、鞘氨醇单胞菌属 (Sphingomonas)、溶杆菌属 (Lysobacter)、假单胞菌属 (Pseudomonas) 等有益菌属以提高其抗病性,恢复细菌多样性和群落稳定性。接种AM真菌可显著降低番茄根际土壤中青枯菌的丰度,特别是侵染青枯菌后提高番茄根际的黄杆菌属 (Flavobacterium)、黄色土源菌属 (Flavisolibacter) 、噬胞菌属 (Cytophaga) 和苔藓杆菌属 (Bryobacter)的相对丰度,进而抑制土壤中青枯菌的生长,并通过提高细菌的多样性和丰富度,促进番茄根际细菌物种之间的稳定共生和良性互作,从而提高细菌群落对青枯菌的抵抗能力。     

施用不同生物有机肥对连作黄瓜枯萎病防治效果及其机理初探

采用营养钵育苗和盆钵试验的方法,研究了以枯草芽孢杆菌SQR9和哈茨木霉T37这两株拮抗菌制成的三种生物有机肥（BIOⅠ、 BIOⅡ和BIOⅢ）对黄瓜生长、 土传枯萎病防治效果及其生防菌株在黄瓜根系中的定殖情况的影响。结果表明, 1）未施用生物有机肥的对照处理（CK）完全发病,有机肥处理（OF）发病率高达88.2%。施用BIO处理都不同程度地降低了黄瓜土传枯萎病的发病率,施用生物有机肥BIOⅠ、 BIOⅡ和BIOⅢ后发病率分别降至51.0%、 19.6%、 13.7%。2）与对照相比,BIOⅠ、 BIO Ⅱ和BIO Ⅲ处理能够显著提高黄瓜生物量,分别是对照的 2.55、 2.46 和 2.58 倍。3）菌株SQR9和T37通过有效的根际定殖降低了病原真菌对根系的侵染,荧光定量PCR（real-time PCR）测定黄瓜根际尖孢镰刀菌数量的结果表明,施用BIOⅡ和BIOⅢ能够将病原菌数量控制在103 copies/g, 土,而对照土壤高达 107 copies/g, 土。     

移栽定殖根际有益菌番茄苗的田间效应研究

为评估移栽定殖根际有益菌(PGPR)番茄苗对果实产量和青枯病防控效果的影响,通过连续3季田间试验,研究了基于生物有机肥的施用,移栽生物育苗基质(在普通育苗基质中添加分离自根际的解淀粉芽孢杆菌)所育种苗(BIONS),相比于移栽普通育苗基质所育种苗(BIO),对设施番茄产量、发病率、收获期植株土体与根际微生物数量和土壤基本理化性质的影响。连续3季田间试验结果表明:相比于BIO处理,BIONS处理第一、二、三季的增产幅度分别达38.86%、47.87%、34.60%,产量差异均达到显著性水平;BIONS处理的发病率每季均极显著低于BIO处理;BIONS处理增加了根际细菌数量,降低了根际真菌数量;土壤基本理化性质方面,BIONS处理的硝态氮含量和铵态氮含量每季均高于BIO处理,且硝态氮含量与产量呈显著正相关,硝态氮含量和铵态氮含量与发病率呈显著负相关。因此,以生物有机肥为底肥,移栽生物育苗基质所育种苗,能够有效防控番茄青枯病的发生,进而提高产量。     

The effect of organic acids from tomato root exudates on rhizosphere colonization of Bacillus amyloliquefaciens T-5

Root colonization by Bacillus amyloliquefaciens is directly related to bacterial growth, chemotaxis, biofilm formation, and the interaction with host plant root exudates. In this study, root exudates were collected from two tomato plant varieties that supported bacterial cell division and induced the B. amyloliquefaciens T-5 chemotactic response, even at the concentration of 10 μg ml⁻¹. Root exudates also induced biofilm formation, but lower than control treatment. In addition, five organic acids were identified in the root exudates and subsequently evaluated. Malic acid, citric acid, succinic acid and fumaric acid significantly induced the chemotaxis response and swarming motility. Maximal chemotactic response and swarming motility were induced by malic acid, and all the organic acid did not have a significant effect on biofilm formation. Furthermore, these organic acids promoted the B. amyloliquefaciens T-5 recruitment under gnotobiotic conditions, increasing the rhizosphere bacterial population. This data suggested that tomato root colonization by B. amyloliquefaciens T-5 was influenced by organic acids secreted by roots. This study expands our understanding of B. amyloliquefaciens T-5 colonization on tomato roots under natural conditions and reflects the significance of B. amyloliquefaciens T-5 strain as biocontrol agent which will be useful for preparing formulations for the better control of plant wilt diseases.     

Rhizobacterium Bacillus amyloliquefaciens Strain SQRT3-Mediated Induced Systemic Resistance Controls Bacterial Wilt of Tomato

Tomato bacterial wilt caused by Ralstonia solanacearum seriously threats tomato growth in tropical and temperate regions around the world. This study reported an antagonistic bacterial strain, Bacillus amyloliquefaciens strain SQRT3, isolated from the rhizosphere soil of tomato plants, which strongly inhibited in vitro growth of pathogenic R. solanacearum. The suppression of tomato bacterial wilt by strain SQRT3 was demonstrated under greenhouse conditions. Additionally, induced systemic resistance (ISR) in tomato as one of the potential disease suppression mechanisms was investigated in the plants inoculated with the isolated bacterial strain SQRT3. The results showed that strain SQRT3 applied with R. solanacearum by drenching significantly reduced tomato bacterial wilt by 68.1% biocontrol efficiency (BE) and suppressed the R. solanacearum populations in the rhizosphere soil compared to the control only drenched with R. solanacearum. The BE of the isolated bacterial strain SQRT3 against tomato wilt increased to 84.1% by root-dipping. Tomato plants treated with both strain SQRT3 and R. solanacearum showed increases in activities of peroxidase and polyphenol oxidase compared with other treatments. The application of strain SQRT3 reduced membrane lipid peroxidation in tomato leaves. The expressions of marker genes for jasmonic acid-and salicylic acid-dependent signaling pathways were faster and stronger in tomato plants treated with both strain SQRT3 and R. solanacearum than in plants treated with either R. solanacearum or strain SQRT3 alone. Collectively, the findings indicated that strain SQRT3 can effectively control tomato wilt.     

The bacterial community of tomato rhizosphere is modified by inoculation with arbuscular mycorrhizal fungi but unaffected by soil enrichment with mycorrhizal root exudates or inoculation with Phytophthora nicotianae

Arbuscular mycorrhizal (AM) fungi have been shown to induce the biocontrol of soilborne diseases, to change the composition of root exudates and to modify the bacterial community structure of the rhizosphere, leading to the formation of the mycorrhizosphere. Tomato plants were grown in a compartmentalized soil system and were either submitted to direct mycorrhizal colonization or to enrichment of the soil with exudates collected from mycorrhizal tomato plants, with the corresponding negative controls. Three weeks after planting, the plants were inoculated or not with the soilborne pathogen Phytophthora nicotianae growing through a membrane from an adjacent infected compartment. At harvest, a PCR-Denaturing gradient gel electrophoresis analysis of 16S rRNA gene fragments amplified from the total DNA extracted from each plant rhizosphere was performed. Root colonization with the AM fungi Glomus intraradices or Glomus mosseae induced significant changes in the bacterial community structure of tomato rhizosphere, compared to non-mycorrhizal plants, while enrichment with root exudates collected from mycorrhizal or non-mycorrhizal plants had no effect. Our results support that the effect of AM fungi on rhizosphere bacteria would not be mediated by compounds present in root exudates of mycorrhizal plants but rather by physical or chemical factors associated with the mycelium, volatiles and/or root surface bound substrates. Moreover, infection of mycorrhizal or non-mycorrhizal plants with P. nicotianae did not significantly affect the bacterial community structure suggesting that rhizosphere bacteria would be less sensitive to the pathogen invasion than to mycorrhizal colonization. Of 96 unique sequences detected in the tomato rhizosphere, eight were specific to mycorrhizal fungi, including two Pseudomonas, a Bacillus simplex, an Herbaspirilium and an Acidobacterium. One Verrucomicrobium was common to rhizospheres of mycorrhizal plants and of plants watered with mycorrhizal root exudates.     

Effect of calcium concentration in nutrient solution on development of bacterial wilt and population of its pathogen Ralstonia solanacearum in grafted tomato seedlings

We examined the effect of the calcium (Ca) concentration in the nutrient solution on the development of bacterial wilt and the population of its pathogen Ralstonia solanacearum in tomato (Lycopersicon esculentum Mill. cv. Momotarou) seedlings grafted onto the rootstock of a highly resistant cultivar (cv. Hawaii 7998). The grafted seedlings were cultured in a nutrient solution containing Ca at concentrations of 0.4, 4.4, and 20.4 mm, and inoculated with the pathogen by stem puncture at the base of the stem of the rootstock, and the disease incidence was recorded for a period of 21 d. In another experiment, xylem exudates were collected from decapitated scions of the Ca-treated seedlings 5 d after inoculation, and the population of the pathogen in the exudates was counted by plating on a selective medium. The grafted tomato seedlings were highly resistant to bacterial wilt, when cultured in the nutrient solution containing Ca at 20.4 mm. The population of the pathogen in the xylem exudates decreased with increasing concentration of Ca in the solution. However, even in the presence of Ca at a high concentration, infection with the virulent pathogen was observed in the xylem of the scion.     

生物有机肥对田间蔬菜根际土壤中病原菌和功能菌组成的影响

利用实时荧光定量PCR方法对田间条件下连作番茄和辣椒施用生物有机肥(BOF)和常规施肥(CK)的根际土壤微生物中青枯病原菌和功能菌群(固氮菌和荧光假单胞菌)的数量进行定量研究.结果表明:与CK相比,BOF处理的番茄和辣椒产量分别提高了26.0％和19.9％,青枯病发病率分别降低了41.5％和44.7％,番茄和辣椒植株根际土壤固氮菌数量分别增加了23.5％和25.8％、荧光假单胞菌数量分别增加了29.5％和20.2％、病原菌数量分别减少了73.2％和90.1％.生物有机肥能够调控根际微生物区系的组成,降低土传病害的发病率,促进作物健康生长;实时荧光定量PCR方法能够快速准确地检测根际土壤中功能微生物种群数量变化.     

Effects of Arbuscular Mycorrhizal Colonization on Microbial Community in Rhizosphere Soil and Fusarium Wilt Disease in Tomato

Fusarium wilt is caused by soil-borne pathogen Fusarium oxysporum. Tomato (Lycopersicon esculentum Mill.) is susceptible to Fusarium oxysporum f. sp. lycopersici race 1 and was infected with wilt disease. A pot experiment was conducted to investigate effects of inoculating arbuscular mycorrhizal (AM) fungus (Glomus etunicatium) on the microbial community in the rhizosphere soil and Fusarium wilt in tomato (cv. Oogatafukuju). The results indicated that AM fungal inoculation suppressed the Fusarium number in the rhizosphere soil of tomato and decreased the Fusarium wilt disease index. Compared to the control, AM fungal inoculation increased the actinomycete number but increased bacterial number. Bacterial and fungal numbers were high but actinomycetes number was low when tomato basal stems became discolored brown. Fusarium inoculation significantly suppressed development of AM colonization and decreased polyphenol oxidase (PPO) activity in leaves and roots of tomato. Inoculation with AM fungi and Fusarium maintained high PPO activity in leaves and roots. The AM colonization increased root growth of tomato, whereas Fusarium inoculation had no significant effect on tomato growth. These findings suggest that because AM fungal inoculation changes microbial communities and enhances PPO activity, it should suppress occurrence of Fusarium wilt in tomato.     

The rhizosphere soil of diseased tomato plants as a source for novel microorganisms to control bacterial wilt

Plant growth promoting rhizobacteria (PGPRs) are used for biocontrol of bacterial wilt caused by Ralstonia solanacearum. They are commonly isolated from the rhizosphere of healthy plants and are scarce in the rhizosphere of diseased plants. We hypothesized that a pathogen-prevalent environment, such as the rhizosphere of infected plants, would be a good or better source for isolating PGPRs than the rhizosphere of healthy plants. In order for these PGPRs to survive successfully in a pathogen-prevalent environment, they must have particularly well-developed survival strategies under the stresses exerted by pathogen activities, which would be of value for their use as biocontrol agents. To test this hypothesis, R. solanacearum-antagonistic bacteria were screened from the rhizospheres of diseased and healthy tomato plants. In total, 110 rhizobacteria were isolated, 18 of which showed antagonism to R. solanacearum in vitro. Among the 18 antagonistic strains, 11 (out of 60) were from the rhizosphere of diseased plants, with inhibition diameter zones ranging from 11.2 to 15.2 mm, whereas 7 (out of 50) were from the rhizosphere of healthy plants, with inhibition diameter zones ranging from 11.5 to 30.5 mm. Strains WR4, WR21, and WR42 from diseased plants rhizosphere, and HR61, HR62, and HR92 from healthy plants rhizosphere, were chosen to investigate their biocontrol efficacies (BCEs) in greenhouse condition. Results showed that WR-isolates performed better in reducing disease incidence (DI) than those HR-isolates. Population densities of R. solanacearum in the rhizosphere soil and crown section of tomato plants were lower in WR-isolate treatments than those in HR-isolate treatments. The best biocontrol effect was achieved by inoculating the strain WR21, followed by WR4, WR42, HR92, HR62, and HR61. Root colonization test showed WR21 had the highest root-colonizing capacity compared with 5 other antagonists. BCEs were positively (r = 0.747) correlated with root-colonizing capacities, but were negatively (r = −0.797) correlated with inhibition zones. In conclusion, the rhizosphere of diseased tomato plants is a good reservoir of biocontrol bacteria.     

Control of tomato bacterial wilt and root-knot diseases by Bacillus thuringiensis CR-371 and Streptomyces avermectinius NBRC14893

Ralstonia solanacearum and Meloidogyne incognita are two soilborne pathogens that cause serious damage and great losses in the production of tomato. For this purpose, a bacterial isolate, Bacillus thuringiensis CR-371, and an actinomyces isolate, Streptomyces avermectinius NBRC14893, were examined for their ability to protect tomato from root-knot nematode and bacterial wilt diseases under glasshouse conditions. Treatment of tomato roots with B. thuringiensis CR-371 and S. avermectinius NBRC14893 followed by challenge inoculation with R. solanacearum and M. incognita significantly decreased disease severity of bacterial wilt alone, root-knot nematode alone, or mixed infection by both pathogens compared to the control. Furthermore, pretreatment of tomato roots with B. thuringiensis CR-371 and S. avermectinius NBRC14893 significantly reduced bacterial proliferation of R. solanacearum both in pathogen alone inoculated plants and in plants co-inoculated with R. solanacearum and M. incognita. In conclusion, our results suggest that the treatment of tomato roots with B. thuringiensis CR-371 and S. avermectinius NBRC14893 simultaneously suppresses bacterial wilt and root-knot nematode diseases. Therefore, B. thuringiensis CR-371 and S. avermectinius NBRC14893 could provide new options for integrated pest management strategies against plant diseases, especially against bacterial-nematode disease complexes that cause synergistic yield losses.