Biological control of bacterial spot of tomato under field conditions at several locations in North America

Following the relatively successful biological control of bacterial speck of tomato under field conditions at several locations (Phytopathology 92 (2002) 1284), similar selection and testing strategies were employed in an effort to isolate an effective biological control agent for bacterial spot of tomato. Fifty potential biological control agents were isolated from tomato foliage in Alabama (AL) and Florida (FL) and tested under greenhouse conditions in AL for the ability to reduce the foliar severity of bacterial spot of tomato (Lycopersicon esculentum), which is caused by either Xanthomonas campestris pv. vesicatoria or Xanthomonas vesicatoria. Three pseudomonads that provided protection against bacterial speck also were included in the tests. The strains which were most efficacious (i.e., high mean percentage reduction) and consistent (i.e., low standard deviation) in reducing bacterial spot severity in repeated greenhouse experiments were selected for field experiments conducted over the period 1996–1998. Among these strains were Cellulomonas turbata BT1, which provided the highest mean reduction in disease severity [45.2% (SD = 21.0)], and Pseudomonas syringae Cit7 [36.4% (SD = 12.2)], which was the most consistent. Field experiments were conducted in Shorter, AL; Bradenton and Sanford, FL; Clinton, North Carolina; Wooster, Ohio; and London, Ontario, Canada. The highest mean reductions in severity of bacterial spot on foliage, averaged across all locations, were provided by P. syringae Cit7 [28.9% (SD = 11.6)] and Pseudomonas putida B56 [23.1% (SD = 7.4)]. The efficacy and consistency of P. syringae Cit7 against bacterial spot were very similar to those achieved against bacterial speck [28.3% (SD = 12.7)] (Phytopathology 92 (2002) 1284). Unfortunately, neither the bacterial strains nor the standard copper bactericides consistently reduced disease incidence on fruit.     

Response of tomato genotypes to bacterial speck (Pseudomonas syringae pv. tomato)

Two genotypes of tomato A 100 and Ontario 7710 which were inoculated separately with four strains of Pseudomonas syringae pv. tomato differed significantly in disease severity (susceptibility) to bacterial speck. At both concentrations of inoculum of each strain used (10⁷ and 10⁸ cfu/ml) A 100 appeared to be highly susceptible whereas Ontario 7710 showed very low or no susceptibility. The significant differences in virulence between strains and in response of tomato plants in three replicate experiments were found. Generally, concentration of inoculum 10⁷ cfu/ml was too low to induce consistent level of disease severity. The obtained results indicate the importance of consistent and favorable conditions for disease development in screening of tomato resistance to bacterial speck.     

Induction of resistance in tomato against cucumber mosaic cucumovirus by plant growth-promoting rhizobacteria

Studies were done to evaluate specific strains of plant growth promoting rhizobacteria (PGPR) for induced resistance against cucumber mosaic cucumovirus(CMV) in tomato. In greenhouse experiments where plants were challenged by mechanical inoculation of CMV, the percentage of symptomatic plants in the most effective PGPR treatments ranged from 32 to 58%,compared with 88 to 98% in the nonbacterized, challenged disease control treatment. Field experiments were conducted in 1996 and 1997 to evaluate 4 PGPR strain treatments based on superior performance in the greenhouse studies. In the 1996field experiment, tomato plants treated with 3 PGPR strains exhibited a significantly lower incidence of CMV infection and significantly higher yields, compared with nonbacterized, CMV-challenged controls. In 1997, the overall percentages of plants infected with CMV in the control and PGPR treatments was higher than in 1996. CMV symptom development was significantly reduced on PGPR-treated plants in 1997compared with the control, but the percentage of infected plants and tomato yields were not significantly different among treatments. These results suggest that PGPR-mediated induced resistance against CMV infection following mechanical inoculation onto tomato can be maintained under field conditions. This revised version was published online in July 2006 with corrections to the Cover Date.     

Application of plant growth-promoting rhizobacteria to control Papaya ringspot virus and Tomato chlorotic spot virus

Papaya ringspot virus (PRSV-W) and Tomato chlorotic spot virus (TCSV) are responsible for severe losses in cucurbits and tomato production in south Florida and other regions in the USA. Traditional chemicals are not effective to control these viruses. Using plant growth-promoting rhizobacteria (PGPR) may present an alternative to control these viruses. Results from this study demonstrated that applying mixtures of PGPR strains is more efficient to control PRSV-W and TCSV compared to individual PGPR strain only. The application method significantly affected the efficiency of PGPR to control PRSV-W and TCSV. The highest reduction in disease severity of both PRSV-W and TCSV occurred in case of soil drenching with PGPR, followed by root dipping and seed coating treatments. Application of PGPR mixtures of (IN937a & SE34) or (IN937a &, SE34 & T4) were the most efficient methods to control these viral diseases.     

Integration of chitosan and plant growth-promoting rhizobacteria to control Papaya ringspot virus and Tomato chlorotic spot virus

Papaya ringspot virus-W (PRSV-W) and Tomato chlorotic spot virus (TCSV) are common viruses infecting vegetables in south Florida. Application of plant growth-promoting rhizobacteria (PGPR) has emerged as a potential alternative of chemical pesticides to control these viruses. But, it is not sufficient to completely replace chemical control. This study aimed to investigate the synergistic effect of chitosan and PGPR to control PRSV-W and TCSV. The efficiency of PGPR to suppress PRSV-W and TCSV was significantly improved when they were accompanied with chitosan treatment. The highest reduction in disease severity of both PRSV-W and TCSV was achieved when chitosan treatment was accompanied with mixture of two PGPR (IN937a + SE34) or three PGPR strains (IN937a + SE34 + SE56). The results of this study proved that implementation of chitosan and PGPR could significantly restrict losses due to PRSV-W and TCSV in squash and tomato, in Florida and the United States.     

抗青枯病型根际促生菌(PGPR)菌群构建及其生物防控机制

【目的】根际促生菌(plant growth promoting rhizobacteria, PGPR)防控青枯病效果不稳定是目前有益微生物生防应用的瓶颈问题,构建稳定高效拮抗青枯菌的PGPR菌群是生物防控的关键。【方法】以前期筛选到的8株PGPR菌株(112、114、Ba-S、TLZZ、LX4、LX7、Ps-S和VC110)和青枯菌(Ralstonia solanacearum, Rs)为研究对象,在前期获得烟草根系分泌物组成的基础上,采用限菌微系统和生态微孔板结合绿色荧光蛋白标记、定量聚合酶链式反应(quantitative polymerase chain reaction, qPCR)、断棒模型设计等方法,探究PGPR菌群对青枯菌入侵烟草根际的抵御机制,并在田间进行抗病、促生效果验证。【结果】LX4、Ba-S、LX7可以充分利用烟草根系分泌物中的氨基酸、糖类碳源抑制青枯菌生长,LX7和112在所有酸类碳源下均对青枯菌有抑制作用,最高抑菌率分别为40.12%(LX7+乳酸)和35.15%(112+柠檬酸)。Ps-S、112和VC110的基础生态位宽度(basal niche b...     

Towards the biological control of fungal and bacterial diseases of tomato using antagonistic Streptomyces spp.

This investigation was designed to explore the potential of microbial antagonism in the control of some tomato diseases including bacterial, Fusarium and Verticillium wilts; early blight; bacterial canker. Three Streptomyces spp. were used: S. pulcher, S. canescens and S. citreofluorescens.The in vitro studies showed that an 80% concentration of the culture filtrate of either S. pulcher or S. canescens significantly inhibited spore germination, mycelial growth and spotulation of Fusarium oxysporum f.sp. lycopersici, Verticillium albo-atrum and Alternaria solani. The same concentration of filtrate of either S. pulcher or S. citreofluorescens was detrimental to the bacterial populations of Clavibacter michiganensis subsp. michiganensis and Pseudomonas solanacearum.The in vivo studies involved different treatments: soaking tomato seeds in filtrate of the antagonist prior to sowing, inoculation of the soil with the antagonist 7 days before sowing, and coating of tomato seeds with spores of the antagonist before sowing. The seed-coating treatment was the most effective in controlling all the pathogens at 42 and 63 days after sowing. Soil inoculation with the antagonist 7 days prior to sowing was less effective in controlling the tomato pathogens as compared to seed-coating. The seed-soaking treatment was the least effective in controlling the diseases concerned.The results also revealed that seed-coating with antagonistic Streptomyces spp. significantly improved tomato growth.     

金银花容器苗对干旱胁迫下接种根际促生细菌的生理响应

在盆栽试验条件下,以金银花容器苗为试材,研究了不同干旱强度下接种蜡样芽孢杆菌(Bacillus cereus)L90对植物生理特征的影响。结果表明:随着干旱胁迫强度的增加,金银花容器苗的光合速率和气孔导度逐渐降低;而干旱环境下接种L90可显著提高气孔导度,缓解干旱胁迫对净光合速率的抑制;且干旱强度增加,缓解效果增强。接种B.cereus L90可显著抑制干旱胁迫下金银花容器苗PSⅡ最大光化学效率、实际光化学效率和光化学猝灭系数的降低,抑制非光化学猝灭系数的升高。虽然L90并没有提高对照处理中光合色素的绝对含量,但可显著抑制干旱环境下金银花叶片中光合色素的分解。干旱显著降低了金银花叶片中细胞分裂素含量,增加了脱落酸(ABA)的含量;在干旱胁迫下,接种L90可显著提高叶片中细胞分裂素的含量,并可促使根部产生的ABA运输到叶片中。干旱胁迫程度较轻时,L90对金银花容器苗的相对含水量和相对电导率影响不显著;而在重度干旱时,同对照相比,干旱及接种L90处理的相对含水量分别降低20.56%和10.21%,相对电导率分别提高31.42%和16.08%,接种L90处理的变化幅度明显较小。因此,干旱生境下接种B.cereus L90,可增加叶片中细胞分裂素含量,抑制光合色素的分解及光合能力的下降,提高金银花容器苗在干旱环境中的适应能力。     

镉胁迫下植物促生菌密歇根克雷伯氏菌TS8和Lelliottia jeotgali MR2对拟南芥生长及镉富集的影响北大核心CSCD

为了研究镉胁迫下植物促生菌密歇根克雷伯氏菌(Klebsiella michiganensis)TS8和Lelliottia jeotgaliMR2对拟南芥(Arabidopsis thaliana)生长及镉富集的影响,文中以野生型拟南芥为试验材料,将其种植在不同镉浓度的土壤基质中,并施入MR2和TS8菌悬液。低浓度镉处理组(LC)为购买的基质营养土,初始镉浓度为14.17 mg/kg,高浓度处理组(HC)为在购买的基质营养土上额外喷洒200 mg/kg Cd^(2+)。结果表明,相比对照组,不同浓度镉胁迫下喷施MR2菌悬液均可显著促进拟南芥的生长,而TS8和MR2_TS8混合菌液仅在高浓度镉胁迫下表现出一定的促生效果。但值得关注的是,不同浓度镉胁迫下TS8菌悬液可显著降低拟南芥的地下部对重金属镉的富集(60%和59%),并有效提高地上部对重金属镉的富集(234%和35%)。此外,单菌和混合菌均能显著提高土壤中可还原态镉向酸可提取态镉转化,促进植物吸收,降低土壤总镉含量。因此,针对不同环境下,合理配施植物促生细菌在提高作物产量或修复土壤镉污染中具有一定的应用价值。     

Metabolic changes in tomato fruits and seeds after viral, bacterial and fungal infections

The metabolic changes in tomato fruits and seeds separately infected with cucumber mosaic virus, Pseudomonas syringae pv. tomato or Botrytis cinerea were investigated cytochemically. The changes of peroxidase (E.C. 1.11.1.7) and β-glucosidase (E.C. 3.2.1.21) were investigated biochemically as well. Tomato fruits were involved in the study because of their high economic value. Tomato seeds were investigated since they have been used most extensively as a model system for studying the physiology and biochemistry of seed development. The diseases caused by the pathogens under study are of special importance for yield reduction in tomato. The three pathogens provoked local changes in the activities of enzymes under study that affect the infected pericarp tissues and neighboring seeds. It was established non-specific and specific responses. The non-specific responses of invaded tissues were expressed as a local enhancement of peroxidase activity in both pericarp tissues and seeds as well as a decrease in activities of: i. enzymes taking part in aerobic and anaerobic respiration, ii. hydrolases esterase and acid phosphatase involved in the basic metabolism as well as an enhancement of their activities in neighboring tissues. Furthermore, it was observed an enhancement of α-galactosidase activity in infected area was observed. The specific responses depending on the type of the pathogen consisted in an enhancement of glucose-6-phosphate dehydrogenase activity by virus infection and an increase of β-glucosidase activity by fungal invasion.     

柽柳根际一株盐单胞菌Bachu 26的分离、鉴定及其盐胁迫下的促生作用研究

盐渍化是世界性的土壤问题,植物促生根际细菌(plant growth-promoting rhizobacteria,PGPR)在盐碱地改良和促进植物生长方面具有独特优势。柽柳是典型的盐生植物,筛选其根际微生物并研究其促生效果与促生机制,以此开发微生物菌肥,具有重要的应用价值。【目的】筛选耐盐碱植物柽柳的根际微生物,对其基本特性、耐盐碱能力、促生功能及促生效果进行评估。【方法】从新疆巴楚境内野生柽柳根际土壤中筛选出一株耐盐碱细菌菌株Bachu 26;通过形态学观察、生理生化特性测定和16S rRNA基因序列分析,对该菌株进行鉴定;利用不同盐浓度(0%–20%)和不同pH(7.0–13.0),对菌株Bachu 26的耐盐耐碱能力进行测定;采用多种功能鉴定培养基测定其促生功能,并对生长素吲哚乙酸(indole-3-acetic acid, IAA)进行定量测定;通过二分格培养皿实验验证菌株产生挥发性酸性物质的能力;在普通培养皿上将拟南芥幼苗与菌株Bachu 26共培养,分析菌株对拟南芥幼苗的促生作用;在二分格培养皿上将拟南芥与Bachu 26隔离培养,分析菌株产生的挥发性酸性物质对拟南芥幼...     

Identification and fine-mapping of a bacterial brown spot disease resistance gene in maize

Bacterial brown spot (BBS) in maize (Zea mays L.) is caused by Pseudomonas syringae pv. syringae Van Holl (Pss). In China, this disease is not prevalent in maize at present. Here, we report the identification and fine mapping of the gene, referred to as Psy1, which confers resistance to BBS. An F₂ population, derived from the cross P25/F349, was used for linkage analysis and mapping of the resistance gene Psy1. Analysis of a BC₈F₂ population, derived from the same parents, confirmed that Psy1 was located on chromosome 10L and inherited as a single dominant gene. For fine mapping of Psy1, two introgression lines, X41 and X44, homozygous at the resistant gene locus, were introduced to hybridize with the susceptible parent P25 respectively, and developed a mixed BC₁ population. We found the closest markers to Psy1 are EST1 and FG29-3, which located on two adjacent BACs respectively, based on the B73 BAC sequence. Sequence analysis of these two BAC sequences (~300 kb) revealed the presence of a homologous sequence of receptor-like kinase. Also a co-segregation marker was developed based on this homologous sequence. These results will be useful for cloning of Psy1 and for transferring or pyramiding Psy1 through MAS in maize breeding programs.     

Three native Pseudomonas fluorescens strains tested under growth chamber and field conditions as biocontrol agents against damping-off in alfalfa

Alfalfa (Medicago sativa) is one of the most important crops used in Uruguay for livestock feeding. Seedling diseases, particularly damping-off, are a critical factor which limits its establishment. Three native Pseudomonas fluorescens strains, UP61.2, UP143.8 and UP148.2, previously isolated from Lotus corniculatus, were evaluated to determine their efficacy as biological control agents for alfalfa seedling diseases in the field. Their compatibility with the alfalfa-Sinorhizobium meliloti symbiosis was also assessed. In growth chamber conditions seed inoculation with Pseudomonas strains did not affect different parameters of alfalfa-rhizobium symbiosis as shown by nodulation rate and shoot dry weight of plants. The presence of the commercial inoculant strains of S. meliloti did not impair colonization by the P. fluorescens and vice versa. In field trials the dynamics of rhizobial rhizospheric populations were not affected by the presence of P. fluorescens. Each P. fluorescens strain successfully colonized alfalfa roots at adequate densities for biocontrol activity. Results showed that P. fluorescens strains provided a 10–13% increase in the number of established plants relative to the control, an intermediate result compared to the fungicide treatment (24%). The alfalfa above-ground biomass was increased by 13% and 15–18% in the presence of the fungicide and P. fluorescens strains, respectively. Therefore, results from this study demonstrated that the three P. fluorescens strains provided effective control against soil-borne pathogens and suggest a potential use in the development of a commercial inoculant to be applied for the control of legume seedling diseases.     

Analysis of pathogenicity mutants of a bacteriocin producing Xanthomonas perforans

Since the initial discovery of Xanthomonas perforans on tomato in 1991, it has completely displaced Xanthomonas euvesicatoria as the bacterial spot of tomato pathogen in Florida. Previous research has shown that X. perforans produces at least three different bacteriocin-like compounds (BcnA, BcnB, BcnC) antagonistic toward X. euvesicatoria strains. In this study pathogenicity-attenuated, bacteriocin-producing mutants of X. perforans were created to determine their potential as biological control agents for control of X. euvesicatoria. Several candidate genes were chosen based on previous studies in which mutant phenotypes exhibited reduced virulence in either X. perforans (OpgH_Xcv) or the closely related X. euvesicatoria strain 85-10 (hpaB, hpaC, xopA, xopD, avrBs2 and gumD). Each candidate gene in X. perforans was amplified and PCR-assisted deletion mutagenesis was performed in the wild-type (wt) X. perforans strain to create potential attenuation mutants. Each mutant was tested for growth rate, disease severity and antagonism toward X. euvesicatoria strains. Three mutants, XopA, opgH, and gumD were significantly less pathogenic than the wild-type strain with the opgH mutant reaching significantly lower internal populations than all other mutants except hpaC. The opgH-strain was the most affected in its ability to grow internally in plant tissue while inhibiting X. euvesicatoria populations equal to or more than the other mutant strains. This mutant strain could potentially be used as part of an effective biological control strategy.     

Spatial and temporal population dynamics of a phyllosphere colonizing Bacillus subtilis biological control agent of sugar beet cercospora leaf spot

This study examines the spatial and temporal variation of populations of the biological control agent (BCA) BacB, a Bacillus subtilis isolate, in the field and growth chamber in the presence of the fungus, Cercospora beticola, the causal agent of Cercospora leaf spot of sugarbeet. The use of the selective BCA support substrate β-glucan, applied at 0, 0.5, and 1.0% of the spray solution, did not influence differences in total population numbers (spores + vegetative cells) of a spontaneous rifampicin resistant isolate of BacB (Rif+) over a 14 day spray period. BacB Rif+, applied as a spore formulation, declined from 10,000 CFU/cm² on day 0.5–100 CFU/cm² on day 14 at the three levels of β-glucan tested. Distribution of BacB Rif+ populations was modeled on a leaf scale, with and without β-glucan. Higher populations of vegetative cells were more likely at 14 days with 1% β-glucan than with 0% β-glucan. BacB populations were more aggregated without β-glucan than with the nutrient substrate. There was no correlation between BacB density and Cercospora leaf spot disease severity, indicating that neither antibiosis nor parasitism is likely an important mechanism of disease control.     

Effect of <Emphasis Type="Italic">Trichoderma</Emphasis> spp. isolates for biological control of tan spot of wheat caused by <Emphasis Type="Italic">Pyrenophora tritici-repentis</Emphasis> under field conditions in Argentina

Trichoderma spp. have been used as biocontrol agents to protect plants against foliar diseases in several crops, but information from field assays is scarce. In the present work, experiments were carried out to determine the effect of six isolates of Trichoderma harzianum and one isolate of T. koningii on the incidence and severity of tan spot, caused by Pyrenophora tritici-repentis (anamorph: Drechslera tritici-repentis) under field conditions. Significant differences between years, wheat cultivars and treatments were found. In 2003, two of the isolates assayed (T5, T7) showed the best performance against the disease applied as seed treatments or sprayed onto wheat leaves at different stages. The application of six of the treatments on wheat plants significantly reduced disease severity by 16 to 35% in comparison with the control. Disease control provided by isolate T7 was similar to that provided by the fungicide treatment (56% reduction). This is the first report on the efficacy of Trichoderma spp. against tan spot under field conditions in Argentina.