Significant Disease Suppression Research Articles

ASSESSMENT OF GARDENING GROWING MEDIA SUPPRESSIVENESS AGAINST RHIZOCTONIA DAMPING-OFF DISEASE

In the present study the suppressive capability was assessed of four commercial peat- based growing media against Rhizoctonia solani on Lepidium sativum. For all organic media, 12 parameters were measured including phytotoxicity, enzymatic activities (glucanase, N- acetylglucosaminidase, chitobiosidase, endochitinase and hydrolysis of fluorescein diacetate), microbiological (culturable bacteria, pseudomonads and fungi) and chemical features (pH, EC and total protein). Three out of all media showed a significant disease suppression, while one was very conducive to disease development. Autoclaving growing media largely decreased or eliminated disease suppressiveness, suggesting that microbes play a vital role. In vitro tests showed a reduction in pathogen suppression when filter-sterilized extracts from the growing media were used. Moreover, disease suppressiveness were positively correlated with EC, supporting the hypothesis of a possible role of physical, other than biological factors, in R. solani control. Our results demonstrate that some peat-based growing media could be useful for the control of soil-borne pathogens.

An antibiotic fusaricidin: a cyclic depsipeptide from Paenibacillus polymyxa E681 induces systemic resistance against Phytophthora blight of red-pepper

In this study fusaricidin, a cyclic depsipeptide isolated from Paenibacillus polymyxa E681 (E681), was demonstrated to control Phytophthora blight infection caused by Phytophthora capsici in red-pepper. The minimal inhibitory concentration (MIC) of fusaricidin was found to be 16 ppm against P. capsici. The disease severity of P. capsici was 40% at 0.1 ppm of fusaricidin when compared with water-treated control (81.7%) on post-treatment, whereas the disease severities on pre-treatment were 45% and 83.3% in fusaricidin (0.1 ppm) and water-treated control, respectively, in red-pepper plants. Significant (P < 0.05) disease suppression was observed on treatment with fusaricidin (0.1 ppm) by foliar spray and soil drench. The disease severity was drastically reduced to 3.3% by soil drench of fusaricidin (1.0 ppm), whereas in water-treated control, the disease severity was 83.3% in the first experiment. Fusaricidin at 0.1 ppm reduced disease severity of P. capsici to 27.5% when compared with positive control (43.1%) and water-treated control (66.2%) in the second experiment. Soft rot disease in tobacco was suppressed upon treatment with fusaricidin at 1.0 ppm by leaf infiltration. RT-PCR analyses of Arabidopsis thaliana revealed that there was an up-regulation of pathogenesis-related (PR) gene expression in wild type A. thaliana (Col-0), while there was no accumulation of PR genes, which implies that the mechanism of protection might be based on a salicylic acid-dependent pathway. This is the first report that fusaricidin exhibits protection against plant pathogens in addition to activity as an antibiotic agent. Hence, E681 can play a role in plant protection by secretion of ISR elicitors including fusaricidin.

Smoke-water Controls Pythium Damping-off in Papaya Seedling

The antifungal efficacy of smoke-water on damping-off caused by Pythium sp. was evaluated both in vitro and in vivo. Smoke-water was generated by burning plant material and bubbling the smoke through water; its effect on the morphology of Pythium sp. was investigated by scanning electron microscope (SEM). Mycelia growth and oospore production of the fungus were significantly inhibited when cultured on water agar amended with smoke-water at 0%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, or 5%. In growth chamber experiments with potted seedlings growing in peatmoss, papaya plants treated with smoke-water exhibited reduced symptoms of damping-off when compared with control plants treated with water. Furthermore, the pots treated with 1.5% or higher smoke-water did not show any symptoms of damping-off disease. Plant height significantly increased with treatment by higher concentrations of smoke-water. Maximum plant height was observed with treatment of 1% smoke-water or higher concentrations. One day after smoke-water application, SEM observations of the Pythium sp. revealed loss of structural integrity, abnormal degradation, deformation, abnormal lysis, cytoplasmic leakage, and hyphal slimming. This study showed that the addition of smoke-water to soil exerted significant disease suppression against Pythium sp., leading to improved growth of papaya seedlings.

Transgenic RNA interference (RNAi)‐derived field resistance to cassava brown streak disease

Cassava brown streak disease (CBSD), caused by the Ipomoviruses Cassava brown streak virus (CBSV) and Ugandan Cassava brown streak virus (UCBSV), is considered to be an imminent threat to food security in tropical Africa. Cassava plants were transgenically modified to generate small interfering RNAs (siRNAs) from truncated full-length (894-bp) and N-terminal (402-bp) portions of the UCBSV coat protein (ΔCP) sequence. Seven siRNA-producing lines from each gene construct were tested under confined field trials at Namulonge, Uganda. All nontransgenic control plants (n = 60) developed CBSD symptoms on aerial tissues by 6 months after planting, whereas plants transgenic for the full-length ΔCP sequence showed a 3-month delay in disease development, with 98% of clonal replicates within line 718-001 remaining symptom free over the 11-month trial. Reverse transcriptase-polymerase chain reaction (RT-PCR) diagnostics indicated the presence of UCBSV within the leaves of 57% of the nontransgenic controls, but in only two of 413 plants tested (0.5%) across the 14 transgenic lines. All transgenic plants showing CBSD were PCR positive for the presence of CBSV, except for line 781-001, in which 93% of plants were confirmed to be free of both pathogens. At harvest, 90% of storage roots from nontransgenic plants were severely affected by CBSD-induced necrosis. However, transgenic lines 718-005 and 718-001 showed significant suppression of disease, with 95% of roots from the latter line remaining free from necrosis and RT-PCR negative for the presence of both viral pathogens. Cross-protection against CBSV by siRNAs generated from the full-length UCBSV ΔCP confirms a previous report in tobacco. The information presented provides proof of principle for the control of CBSD by RNA interference-mediated technology, and progress towards the potential control of this damaging disease.

Systemic resistance induced in Arabidopsis thaliana by Trichoderma asperellum SKT‐1, a microbial pesticide of seedborne diseases of rice

Trichoderma asperellum SKT-1 is a microbial pesticide of seedborne diseases of rice. To investigate the mechanisms of disease suppression in SKT-1, the ability to induce systemic resistance by SKT-1, or its cell-free culture filtrate (CF), was tested using Arabidopsis thaliana Col-0 plants. Both SKT-1 and its CF elicit an induced systemic resistance against the bacterial leaf speck pathogen Pseudomonas syringae pv. tomato DC3000 in Col-0 plants. Involvement of plant hormones in the induced resistance by SKT-1 and CF was assessed using Arabidopsis genotypes such as the jasmonic acid (JA)-resistant mutant jar1, the ethylene (ET)-resistant mutant etr1, the plant impaired in salicylic acid (SA) signalling transgenic NahG and the mutant npr1 impaired in NPR1 activity. In soil experiments using SKT-1, no significant disease suppression effect was observed in NahG transgenic plants or npr1 mutant plants. Expression levels of SA-inducible genes such as PR-1, PR-2 and PR-5 increased substantially in the leaves of Col-0 plants. Expression levels of JA/ET-induced genes such as PDF1.2a, PR-3, PR-4 and AtVsp1 were also induced, but the levels were not as high as for SA-inducible genes. In a hydroponic experiment using CF from SKT-1, all Arabidopsis genotypes showed an induced systemic resistance by CF and increased expression levels of JA/ET- and SA-inducible genes in leaves of CF-treated plants. The SA signalling pathway is important in inducing systemic resistance to colonisation by SKT-1, and both SA and JA/ET signalling pathways combine in the signalling of induced resistance by CF. These results indicate that the response of A. thaliana is different from that found in root treatments with barley grain inoculum and CF from SKT-1.

Suppression of autoimmunity and organ pathology in lupus‐prone mice upon inhibition of calcium/calmodulin‐dependent protein kinase type IV

Systemic lupus erythematosus (SLE) is a chronic inflammatory disease associated with aberrant immune cell function. Treatment involves the use of indiscriminate immunosuppression, which results in significant side effects. SLE T cells express high levels of calcium/calmodulin-dependent protein kinase type IV (CaMKIV), which translocates to the nucleus upon engagement of the T cell receptor-CD3 complex and accounts for abnormal T cell function. The purpose of this study was to determine whether inhibition of CaMKIV would improve disease pathology. We treated MRL/lpr mice with KN-93, a CaMKIV inhibitor, starting at week 8 or week 12 of age and continuing through week 16 and evaluated skin lesions, proteinuria, kidney histopathology, proinflammatory cytokine production, and costimulatory molecule expression. We also determined the effect of silencing of CAMK4 on interferon-γ (IFNγ) expression by human SLE T cells. CaMKIV inhibition in MRL/lpr mice resulted in significant suppression of nephritis and skin disease, decreased expression of the costimulatory molecules CD86 and CD80 on B cells, and suppression of IFNγ and tumor necrosis factor α production. In human SLE T cells, silencing of CAMK4 resulted in suppression of IFNγ production. We conclude that suppression of CaMKIV mitigates disease development in lupus-prone mice by suppressing cytokine production and costimulatory molecule expression. Specific silencing of CAMK4 in human T cells results in similar suppression of IFNγ production. Our data justify the development of small-molecule CaMKIV inhibitors for the treatment of patients with SLE.

Genetic and molecular characterization of a blue light photoreceptor MGWC-1 in Magnaporth oryzae

Three key factors involved in successful plant disease development include the presence of a susceptible host, a virulent pathogen, and a disease-conducive environment. Our understanding of how environmental factors influence disease-conducive or disease-suppressive conditions, and how a pathogen advantageously capitalizes on them, is quite limited. Utilizing the model pathosystem Magnaporthe oryzae– Oryza sativa, we found a significant light-dependent disease suppression. Our genetic data suggest that the blue-light receptor MGWC-1 in M. oryzae is involved in light-dependent disease suppression during the dark-phase (disease-conducive light condition) immediately after pathogen–host contact. Sensing “darkness” is accomplished by MGWC-1, a blue-light receptor in M. oryzae. To explore the potential molecular mechanisms of light-dependent disease suppression we performed a genome-wide microarray experiment and identified several groups of gene families that are differentially regulated during the light-to-dark transition. Our genetic and molecular data provide insights into how a fungal pathogen utilizes ambient light signals for successful disease development.

Selection of genetically diverse Trichoderma spp. isolates for suppression of Phytophthora capsici on bell pepper

Environmentally compatible control measures are needed for suppression of Phytophthora capsici on pepper. Twenty-three isolates of Trichoderma were screened for suppression of a mixture of 4 genetically distinct isolates of this pathogen on bell pepper (Capsicum anuum) in greenhouse pot assays. Of these 23 isolates, GL12, GL13, and Th23 provided significant suppression of P. capsici in at least 2 assays. These isolates were then compared with Trichoderma virens isolates GL3 and GL21 for suppression of this disease in the presence and absence of the harpin-based natural product Messenger. Isolates GL3 and Th23 provided significant disease suppression (P ≤ 0.05) in 3 of 4 assays, while GL12, GL13, and GL21 provided significant suppression in 2 of 4 assays. There was no apparent benefit from the application of Messenger. Phylogenetic analysis of these 5 isolates (based on the ITS1 region of the nuclear rDNA cluster and tef1), and an additional 9 isolates that suppressed P. capsici in at least 1 assay, separated isolates into 2 clades, with 1 clade containing GL3, GL12, GL13, and GL21. There were also 2 more distantly related isolates, one of which was Th23. We report here the identification of genetically distinct Trichoderma isolates for potential use in disease management strategies employing isolate combinations directed at suppression of P. capsici on pepper.

Immune regulation: FcγRIIB—regulating the balance between protective and autoreactive immune responses

Immune regulation: FcγRIIB—regulating the balance between protective and autoreactive immune responses

Suppression of Pythium ultimum root rot of sorghum by rhizobacterial isolates from Ethiopia and South Africa

Bacteria isolated from the rhizosphere of sorghum in Ethiopia and from different grass species within the Nylsvlei Nature Reserve in South Africa were evaluated for their in-vitro and in-vivo antagonistic activity against Pythium ultimum that causes root rot in sorghum. Root rot caused by this pathogen in sorghum is characterized by blackened discolorations with white mycelial growth on the surface of the roots. Statistically significant disease suppression was achieved by 15 isolates from sorghum rhizosphere in Ethiopia (⩾50% disease suppression), whereas eight isolates from the rhizosphere of grasses at Nylsvlei Nature Reserve in South Africa resulted in ⩾80% disease suppression. The isolates maintained themselves at a level ⩾105cfu/g indicating that they are also highly competent in sorghum rhizosphere. Isolates which showed the best overall performance in-vitro and in-vivo were identified by means of the API system and sequencing of the bacterial 16S rDNA genes. The majority of these isolates were identified as members of the genus Bacillus, 56% of which were B. cereus. Three other isolates were identified as Brevibacterium laterosporus, Pseudomonas fluorescens and Serratia marcescens. Selected modes of action studies indicated that production of antibiotic substances and siderophores as well as induction of systemic resistance in sorghum were exhibited by some of the most effective isolates. The results of this study offer a significant impetus to the application of plant growth promoting rhizobacteria for use as biological control agents against P. ultimum in sorghum.

Sequential selection and efficacy of antagonistic rhizobacteria for controlling Phytophthora blight of pepper

This research was conducted to develop a sequential screening procedure for bacteria antagonistic against Phytophthora capsici, and to evaluate control efficacy by candidate antagonistic bacteria selected by this procedure against Phytophthora blight of pepper in the field. A total of 231 bacterial strains were isolated from soils and roots of cucumber, pepper, and tomato plants grown in fields at 12 locations in Korea in 2000 and 2001. From these, 88 strains were pre-selected using a radicle assay. These strains were then screened in a seedling assay with 2-week-old pepper seedlings inoculated with 1000 zoospores of P. capsici g −1 potting mixture, a concentration which caused complete death of the seedlings. Based on the results of this screen, 15 potentially antagonistic strains with at least 50% control efficacy of seedling infection by P. capsici were selected and further tested in 5-week-old pepper plants. In the in planta trials, four bacterial strains, KJ1R5, KJ2C12, KJ9C8, and 11S16, consistently showed significant ( P=0.05) disease suppression against P. capsici infection and were examined as candidate biocontrol agents for their antagonistic effects in the field by artificial inoculation in 2001 and 2002. Plants treated with the bacterial strains KJ1R5, KJ2C12, and KJ9C8 had significantly ( P=0.05) lower disease severity and incidence than untreated controls in at least one test. These field test results have demonstrated that the radicle and seedling assays are very effective as part of the screening procedure to select candidate antagonistic bacteria against P. capsici on pepper. Therefore, effective biocontrol agents for Phytophthora blight of pepper could be selected through the sequential screening procedure developed in this study.

Suppression of established experimental autoimmune uveitis by anti-LFA-1alpha Ab.

To identify costimulatory molecules that are important in the effector phase of experimental autoimmune uveitis (EAU). EAU was induced in C57BL/6 (B6) mice by transfer of activated T cells specific for the interphotoreceptor-binding protein (IRBP) 1-20 peptide. The animals were then treated with and without anti-leukocyte function associated antigen (LFA)-1alpha mAb, at day 0 or 10 (disease onset) after T-cell transfer. Clinical signs of inflammation, ocular histology, and infiltrated inflammatory cells in the eye were compared. The primary and secondary proliferative responses of uveitogenic CD4 and CD8 T cells were tested after treatment with costimulatory blockers in vivo and in vitro. Moreover, the abilities of uveitogenic T cell trafficking and their interaction with retinal astrocytes were examined. Anti-LFA-1alpha Abs caused significant suppression of disease when administered either at the time of effector uveitogenic T cell transfer or at disease onset. Studies of the mechanisms by which anti-LFA-1alpha Ab inhibits the effector phase of uveitis demonstrated that it blocks multiple pathogenic events of uveitis mediated by IRBP-specific uveitogenic T cells, including the activation of T cells outside and inside the eye and the trafficking of activated autoreactive T cells into the inflammatory site. In addition, Ab treatment selectively suppressed the activation and expansion of pathogenic, but not regulatory, T cells in vivo. Anti-LFA-1alpha Abs are potent inhibitors of established autoimmune uveitis and that such treatment may be applicable not only for the prevention, but also the treatment, of T-cell-mediated autoimmune diseases.

The protease inhibitor, Bowman-Birk inhibitor, suppresses experimental autoimmune encephalomyelitis (131.33)

Abstract The Bowman-Birk inhibitor (BBI) is a soybean-derived serine protease inhibitor. BBI concentrate (BBIC) is enriched with BBI, but is predominantly made up of other ingredients. In our previous study we found that oral administration of BBIC to Lewis rats with EAE resulted in significant clinical and histological suppression of disease. In the present study we investigated the effect of BBI using mouse EAE models and pure BBI. We found that both I.P. and oral treatment with BBI (typically 1 mg/day BBI) in SJL/PLP139–151, or C57BL/6/MOG35–55 models, significantly improved clinical and histological parameters of EAE (disease onset, severity, weight loss, inflammation and demyelination). This was true for different treatment regimens as regards the day of treatment initiation relative to immunization for EAE induction. In most experiments antigen-specific proliferation of immune cells derived from BBI-treated mice was significantly lower relative to control groups. In the SJL model of optic neuritis BBI significantly reduced the incidence of neuritis, inhibited inflammation and prevented loss of retinal ganglion cells. Using Boyden’s chamber assay we found that BBI inhibited invasiveness of activated splenocytes through the matrigel barrier. These results indicate that BBI is an excellent candidate for oral therapy in multiple sclerosis.

Bacillus vallismortis Strain EXTN-1 Mediated Systemic Resistance against Potato virus Y and X in the Field

Efficacy of plant growth promoting rhizobacteria(PGPR) Bacillus vallismortis strain EXTN-1 has been proved in eliciting induced systemic resistance(ISR) in several crops. The present paper described the beneficial effects of EXTN-1 in potato as increase in yield and chlorophyll content, and plant protection against Potato Virus Y and X(PVY & PVX). EXTN-1 induced systemic resistance to the plants resulting in significant disease suppression in the field. Also the plants under treatment with EXTN-1 had higher chlorophyll content. The bacterized plants had significantly higher yields over the untreated control plants. The strain induced activation of defense genes, PR-1a and PDF 1.2 in transgenic tobacco model, which indicated the possible role of both SA, and JA pathways in EXTN-1 mediated plant protection against crop diseases.

Variability Associated with Suppression of Gray Mold (Botrytis cinerea) on Geranium by Foliar Applications of Nonaerated and Aerated Compost Teas.

The use of compost teas in agriculture and urban landscapes is increasing, but there is conflicting information on which methods are suitable for obtaining compost teas that suppress disease when applied to plants. Numerous compost tea production parameters (compost source, aeration, nutrient additives, production duration, and use of application adjuvants) were tested for suppressing gray mold (Botrytis cinerea) on geranium. While individual compost tea batches did significantly reduce disease, the majority of compost teas did not significantly suppress gray mold of geranium. For nonaerated compost tea (NCT), the most consistent, significant disease suppression was associated with particular compost samples and increased production time, with little effect of periodic stirring or the addition of nutrients at the onset of production. Continuously aerating compost tea did not significantly increase disease suppression compared with nonaerated compost teas. Preparing aerated compost tea (ACT) with nutrient additives did not consistently increase disease suppression; however, 67% of ACT batches made with a mixture of kelp extract, rock dust, and humic acid significantly reduced disease. Applying ACT with spray adjuvants significantly reduced disease compared with ACT with no adjuvants. The variability in gray mold suppression from NCT and ACT applications indicates that disease control would not be commercially acceptable unless there are no other alternatives due to production constraints, e.g., organic standards.

Preliminary phenotypic and molecular screening for potential bacterial biocontrol agents of Leptosphaeria maculans, the blackleg pathogen of canola

Leptosphaeria maculans causes blackleg disease of canola (Brassica napus L.). Bacteria isolated from soil, canola stubble and plant parts were assayed for suppression of blackleg. In plate assays, the bacteria isolated from canola stubble had the highest agar-diffusible antifungal activity (75%), which was fungitoxic. In plant cotyledon assays, endophytes had the highest disease suppression. Bacteria with the highest disease suppression in cotyledon assays also had significant disease suppression at the three- to four-leaf stage. PCR screening for bacterial biosynthetic genes, commonly thought to be involved in plant disease suppression, revealed 22 bacteria to be positive for pyrrolnitrin. Pseudomonas chlororaphis and P. aurantiaca isolates contained the phenazine biosynthetic gene. Three Bacillus cereus isolates had the zmaR resistance gene. This study generated a novel set of primers specific to the zwittermicin A biosynthetic cluster. The PCR screening has confirmed the presence of genes encoding pyrrolnitrin (55%), phenazine (10%), zwittermicin A biosynthesis (7.5%) and zwittermicin A resistance (7.5%) from the canola phyllosphere and rhizosphere, which seems more widely distributed than genes for 2,4-diacetylphloroglucinol and pyoluteorin.

Suppressiveness of 18 composts against 7 pathosystems: Variability in pathogen response

Compost is often reported as a substrate that is able to suppress soilborne plant pathogens, but suppression varies according to the type of compost and pathosystem. Reports often deal with a single pathogen while in reality crops are attacked by multiple plant pathogens. The goal of the present study was to evaluate the disease suppression ability of a wide range of composts for a range of plant pathogens. This study was conducted by a consortium of researchers from several European countries. Composts originated from different countries and source materials including green and yard waste, straw, bark, biowaste and municipal sewage. Suppressiveness of compost-amended (20% vol./vol.) peat-based potting soil was determined against Verticillium dahliae on eggplant, Rhizoctonia solani on cauliflower, Phytophthora nicotianae on tomato, Phytophthora cinnamomi on lupin and Cylindrocladium spathiphylli on Spathiphyllum sp., and of compost-amended loamy soil (20% vol./vol.) against R. solani on Pinus sylvestris and Fusarium oxysporum f. sp. lini on flax. From the 120 bioassays involving 18 composts and 7 pathosystems, significant disease suppression was found in 54% of the cases while only 3% of the cases showed significant disease enhancement. Pathogens were affected differently by the composts. In general, prediction of disease suppression was better when parameters derived from the compost mixes were used rather than those derived from the pure composts. Regression analyses of disease suppression of the individual pathogens with parameters of compost-amended peat-based mixes revealed the following groupings: (1) competition-sensitive: F. oxysporum and R. solani/cauliflower; (2) rhizosphere-affected: V. dahliae; (3) pH-related: P. nicotianae; and (4) specific/unknown: R. solani/pine, P. cinnamomi and C. spathiphylli. It was concluded that application of compost has in general a positive or no effect on disease suppression, and only rarely a disease stimulating effect.

Biofumigation and Soil Conditioning Effects of Cover Crops in Processing Tomato

Mustard cover crop residue has been reported to have a “biofumigant” action when incorporated into the soil, potentially providing significant disease suppression and yield improvement for the succeeding crop. Such activity could be particularly useful in processing tomato rotations, where consecutive cropping invariably results in yield decline. Agronomic and environmental effects of growing over-winter mustard cover crops preceding tomato production were investigated in three field trials between 2002 and 2004. Two mustard cover crops [`Pacific Gold', a brown mustard (Brassica juncea), and `Caliente', a blend of brown and white mustard (Sinapis alba)] were compared to a legume cover crop mix, a fallow bed treatment (the standard grower practice in this region), and, in two of the three trials, a fumigation treatment using metam sodium. No suppression of soil populations of Verticillium dahliae or Fusarium spp. was observed with the mustard cover crops, nor was there any visual evidence of disease suppression on subsequent tomato crops. In these fields, the mustard either had no effect, or reduced tomato yield, when compared to the fallow treatment. At one of two sites, metam sodium fumigation significantly increased tomato yield. The presence of a cover crop, whether mustard or legume, reduced winter runoff by an average of 50% over two years of trials. No benefit of mustard cover cropping beyond this reduction in winter runoff was observed.

EFFICACY OF BIOLOGICAL AMENDMENTS ON POSTTRANSPLANT ROOT ROT

Biological substrate amendments including SG-11, Subtilex, SoilGuard, ActinoIron, Companion, RootShield and BioYield were evaluated for their efficacy to control common soil-borne fungal and fungal-like pathogens when incorporated into the substrate at transplanting. The biological agents were incorporated into an 80% Sphagnum peat and 20% perlite substrate at the label recommended rates and four-to-six-leaf plugs of the test species were transplanted into the substrates. Substrates were either inoculated or uninoculated with a test pathogen. Pathogen-host combinations included Pythium ultimum on geranium (Pelargonium ×hortorum), Phytophthora nicotianae and Pythium aphanidermatum on vinca (Catharanthus roseus), and Theilaviopsis basicoli on pansy (Viola ×wittrockiana). The incidence of disease development, plant mortality and root fresh weights did not differ among the biological agents and the inoculated controls. Therefore, under the conditions of this study, the biological agents did not provide significant disease suppression. Pansy and vinca plants grown in uninoculated substrates amended with Subtilex had significantly higher shoot dry weights than those grown in unamended substrates. Pansy, vinca and tomato plants grown in uninoculated substrates amended with SG-11 had significantly higher shoot dry weights than those grown in unamended substrates.

Regulatory functions of CD8+CD28- T cells in an autoimmune disease model.

CD8+ T cell depletion renders CD28-deficient mice susceptible to experimental autoimmune encephalomyelitis (EAE). In addition, CD8-/-CD28-/- double-knockout mice are susceptible to EAE. These findings suggest a role for CD8+ T cells in the resistance of CD28-deficient mice to disease. Adoptive transfer of CD8+CD28- T cells into CD8-/- mice results in significant suppression of disease, while CD8+CD28+ T cells demonstrate no similar effect on the clinical course of EAE in the same recipients. In vitro, CD8+CD28- but not CD8+CD28+ T cells suppress IFN-gamma production of myelin oligodendrocyte glycoprotein-specific CD4+ T cells. This suppression requires cell-to-cell contact and is dependent on the presence of APCs. APCs cocultured with CD8+CD28- T cells become less efficient in inducing a T cell-dependent immune response. Such interaction prevents upregulation of costimulatory molecules by APCs, hence decreasing the delivery of these signals to CD4+ T cells. These are the first data establishing that regulatory CD8+CD28- T cells occur in normal mice and play a critical role in disease resistance in CD28-/- animals.