Extracellular self-DNA plays a role as a damage-associated molecular pattern (DAMP) delaying zoospore germination rate and inducing stress-related responses in Phytophthora capsici

Phytophthora capsici is a highly destructive pathogen of crops. Although chemical pesticides are the most widely used strategy to counter phytopathogens, they have been inefficient to combat P. capsici and have produced significant environmental and health problems. Therefore, sustainable alternatives to control soilborne pathogens, such as the inhibitory effect of self-extracellular DNA (eDNA), have been proposed. This inhibition phenomenon has been attributed to the action of self-eDNA as a damage-associated molecular pattern (DAMP). Here, we describe the effect of self-eDNA on P. capsici zoospore germination rate, antioxidant enzymes activity and MAPK gene expression. Also, the effect of P. capsici eDNA on the protection of chilli pepper (Capsicum annuum) plants against P. capsici was investigated. The results highlight that P. capsici can sense 2–500 µg/ml self-eDNA and induce stress-related responses like SAK1 gene expression, and superoxide dismutase and catalase activities. Moreover, in vitro zoospore germination rate was suppressed with self-eDNA concentrations ranging from 50 to 500 µg/ml. Interestingly, drench applications of P. capsici eDNA at 60 and 100 µg/ml on chilli pepper plants did not show any protective effect against the phytopathogen, whereas 2 µg/ml of P. capsici eDNA drench application showed a lower percentage of plants with symptoms and lower disease severity. Moreover, phenols and total flavonoids were increased in chilli pepper plants, therefore inducing plant immunity. This study showed that self-eDNA acts as a DAMP in P. capsici and provides insight into the use of eDNA for the protection of crops of agronomic interest.     

Effect of high-power monochromatic (pulsed UV laser) and low-power broadband UV radiation on Phytophthora spp. in irrigation water

Cysts or zoospores of Phytophthora capsici, P. citrophthora and P. nicotianae were suspended in distilled water or in recycled irrigation water collected from commercial nurseries. Propagules, suspended in shallow layers of still water in 60 mm-diameter plastic Petri dishes with lids off, were exposed to various UV doses emitted from either an excimer laser or a mercury vapor lamp. The laser emitted a monochromatic (248 nm, 5 eV/pulse), pulsed UV beam with an intensity of 1 to 2 mJ/cm²/pulse with 20-ns pulse durations and a high peak power of 50 to 100 kW/cm²/pulse. The UV lamp was from a commercial water purifier that emitted broadband (continuous) UV radiation at an intensity of ～8 mW/cm². Survival was assessed by culturing aliquots of the treated or non-treated suspensions onto corn meal agar amended with ampicillin (250 ppm) and rifampin (10 ppm), and then counting numbers of developing colonies. The UV dose (as energy per unit area) required to kill propagules was smaller when they were suspended in distilled water than when suspended in recycled nursery water. The reduced kill effectiveness in recycled water appears to be related to UV-absorbing soluble chemicals. Cysts and zoospores were equally susceptible to UV, although the high-peak power pulsed-UV laser source with ultra-short exposure times appeared to have greater kill effectiveness than the conventional Hg-vapor UV lamp. Phytophthora capsici and P. nicotianae isolates were somewhat less sensitive to UV than isolates of P. citrophthora obtained from various hosts and geographical regions. Furthermore, hyphae of Phytophthora spp. were less susceptible to UV than were cysts or zoospores.     

Activation of tomato plant defence responses against bacterial wilt caused by Ralstonia solanacearum using DL-3-aminobutyric acid (BABA)

In this study, we investigated the ability of DL-3-aminobutyric acid (BABA) to protect tomato against bacterial wilt caused by Ralstonia solanacearum. This was combined with studies of accumulation of total phenolic compounds, free and total salicylic acid (SA), and activity of enzymes related to plant defence, i.e., polyphenol oxidase (PPO) and catalase (CAT). Under greenhouse conditions, tomato plants pre-treated by soil drenching with BABA profoundly reduced disease severity of bacterial wilt compared to plants receiving a soil drench with water. Thus, BABA reduced leaf wilting index by 75.3 % and vascular browning index by 69.9 %, without any in vitro inhibitory activity on the pathogen. BABA treatment significantly reduced the population of R. solanacearum in stems of tomato plants and additionally also significantly increased both fresh and dry weight of roots and shoots of tomato plants compared with the inoculated control. Application of BABA resulted in a high increase in PPO activity both in plants with and without inoculation. Compared to water-treated plants, treatment with BABA also induced a significant increase of total phenolic compounds as well as of free and total SA in leaves of both inoculated and non-inoculated tomato plants at all sampling times. CAT activity decreased in tomato plants treated with BABA in comparison with the water-treated control plants and the decrease in activity correlated with an increasing total SA accumulation. These findings suggest that BABA treatment resulted in induction of resistance to bacterial wilt in tomato.     

Direct antifungal activity of tiadinil,a systemic acquired resistance inducer,and thymol formulations on <Emphasis Type="Italic">Stagonosporopsis citrulli</Emphasis> and control of watermelon gummy stem blight

This study evaluated the direct antifungal activity of tiadinil [N-(3-chloro-4-methylphenyl)-4-methyl-1,2,3-thiadiazole-5-carboxamide], a systemic acquired resistance (SAR) inducer and two formulations of thymol (thymol I and thymol II) against Stagonosporopsis citrulli, the causal agent of gummy stem blight (GSB) disease of watermelon. Tiadinil, thymol I and thymol II completely inhibited the mycelial growth of S. citrulli in vitro at ≥?100 ppm. Conidial germination and germ tube elongation were completely inhibited by tiadinil at ≥?2000 ppm and by thymol-based formulations at ≥?100 ppm. A single foliar application of tiadinil at ≥?10 ppm or a single application of thymol I and II at ≥?1 ppm, 48 h before or after pathogen inoculation, significantly reduced disease severity of watermelon seedlings inoculated with 10⁵/ml conidial suspension of S. citrulli, compared to respective nontreated controls. Plants treated with foliar application of tiadinil at ≥?1000 ppm before pathogen inoculation had significantly lower disease severity than plants that received an equivalent drench application. The efficacy of foliar application of tiadinil was affected by concentration and frequency of application. The study suggests direct antifungal activity of tiadinil, indicating a new mode of action of tiadinil against GSB disease of watermelon. The study also demonstrated direct antifungal action of thymol, a formulated active compound of essential oils, against S. citrulli and GSB disease of watermelon.     

<Emphasis Type="Italic">Aphelenchoides gorganensis</Emphasis> n. sp. (Nematoda: Aphelenchoididae)<Emphasis Type="Italic">,</Emphasis> a new species from Iran

Phytophthora capsici infection of chili pepper seedlings can cause substantial losses due to damping-off and collar rot diseases. Chemical control is no longer effective due to reported resistance development, on top of the related environmental concerns and the consumer demands for reduced use of fungicides. Biological control is a sustainable option, with several agents having been reported to be effective against this pathogen. This research focused on optimizing the application of strain THSW13 of Trichoderma hamatum and a bacterial isolate BJ10–86 with the objectives of improving chili pepper seed germination, reduce damping-off disease incidence, and improve the growth of the seedlings. Bacterial isolate BJ10–86 was subjected to molecular identification and found to be Pseudomonas aeruginosa. Chili pepper seeds treated with the biocontrol agents, individually or in combination, were seeded into commercial nursery media that had been pre-inoculated with P. capsici zoospores. Over a period of 35 days the chili pepper seed treatments significantly (P = 0.008) reduced the disease incidence of seedlings damping-off. Combined application of T. hamatum and P. aeruginosa was the best biocontrol treatment with an area under disease curve of only 36.61 units compared to 92.87 units for the control treatment. Similar results were observed in vitro where T. hamatum and P. aeruginosa synergistically inhibited P. capsici growth by 73.2 %. The inhibition activity of this treatment was similar to mefenoxam treatment, which implies that it is an effective and sustainable alternative for chili pepper seed treatment. The biocontrol seed treatment had no effect on seed germination and seedling growth.     

Induced resistance againstPhytophthora capsici in pepper plants in response to DL-β-amino-n-butyric acid

Treatment of pepper plants with the nonprotein amino acid, DL-ß-amino-n-butyric acid (BABA) induced resistance to subsequent infection byPhytophthora capsici. In contrast, theα-, andγ-isomers of aminobutyric acid were ineffective as inducers of resistance. A relatively high concentration of BABA at 1,000μg ml^?1, which had no antifungal activityin vitro againstP. capsici, was required to induce resistance against Phytophthora blight with a foliar and stem spray, thus leading to complete control of the disease. About 1 day interval between BABA-treatment and challenge inoculation was sufficient to induce resistance in pepper plants. High inoculum levels ofP. capsici caused Phytophthora development slowly in pepper stems treated with BABA, especially at early plant growth stage, which suggests that the induced resistance in pepper plants may be more quantitative rather than qualitative. BABA applied to the root system also protected pepper stems fromP. capsici infection.     

Modified plant architecture to enhance crop disease control: genetic control and possible value of upright fruit position in cucumber

Plant architectural features can facilitate disease control by creating unfavourable environments for pathogen growth or limiting pathogen contact with the host. In the case of Phytophthora capsici infection of cucumber (Cucumis sativus), the susceptible fruit typically lie in contact with soil under warm, moist conditions of a full canopy, an ideal location for exposure to inoculum and disease development. We have shown that increased row spacing, trellising and architectural variants that allow for more open canopies, such as shorter vines, reduced branching, or smaller leaves, can modify microclimate as assessed by temperature at the location of developing fruits. However, only trellising reduced infection rates (<3 % infection vs. 22 % for control). A cucumber accession PI308916, with extremely short internodes and resulting upright position of young fruit, also exhibited reduced disease. Fruit of PI308916 were susceptible when inoculated with P. capsici, indicating that reduced infection likely resulted from decreased soil contact. The compact trait is inherited as a single recessive gene, cp. Like many architectural traits, cp may be pleiotropic; PI308916-derived breeding lines were reported to have poor seedling emergence. PI308916 exhibited inconsistent apical hook formation that co-segregated with short internodes in F₂ and backcross generations, and disorganized internodal cortical cell arrangement. The combined phenotypes suggest either pleiotropy or very tight linkage of genes affecting hormone balance or cell division. The narrow chromosome region recently defined by QTL analysis includes candidate gene homologs of meristem related genes modulating cell division or spacing of lateral shoot organs and genes associated with brassinosteroid signaling or cytokinin content.     

Soil amendments with Brassica cover crops for management of Phytophthora blight on squash

BACKGROUND: Phytophthora blight induced by Phytophthora capsici is responsible for serious yield loss in vegetable production in the United States and other countries. This study was conducted to evaluate the efficacy of Brassica cover crops used as soil amendments for managing Phytophthora blight of squash. RESULTS: In greenhouse studies, disease incidence on squash plants was significantly reduced by soil amendment with mustard shoots or roots used at 1 and 2.5% (plant tissue/soil, w/w). The shoots of canola used at 1 or 2.5% also suppressed disease, while the roots of canola or other crops did not reduce disease significantly. In field studies, soil amendments with mustard and canola provided the greatest disease reduction and increased squash yield significantly compared with the non‐treated control. Mustard and canola did not appear to be susceptible to P. capsici. CONCLUSION: The results indicated that some Brassica crops, particularly mustard and canola, had the potential to significantly reduce Phytophthora blight on squash when used as soil amendments. As P. capsici has a remarkable ability to develop resistance to chemical fungicides, use of effective Brassica cover crops could be a biorational alternative to fungicides and a valuable component in developing integrated disease management programs. Copyright © 2011 Society of Chemical Industry     

Inhibition of plant pathogens in vitro and in vivo with essential oil and organic extracts of Cestrum nocturnum L.

The efficacy of the essential oil and various organic extracts from flowers of Cestrum nocturnum L. was evaluated for controlling the growth of some important phytopathogenic fungi. The oil (1000 ppm) and the organic extracts (1500 μg/disc) revealed antifungal effects against Botrytis cinerea, Colletotrichum capsici, Fusarium oxysporum, Fusarium solani, Phytophthora capsici, Rhizoctonia solani and Sclerotinia sclerotiorum in the growth inhibition range of 59.2-80.6% and 46.6-78.9%, respectively, and their MIC values were ranged from 62.5 to 500 and 125 to 1000 μg/mL. The essential oil had a remarkable effect on spore germination of all the plant pathogens with concentration and time-dependent kinetic inhibition of P. capsici. Further, the oil displayed remarkable in vivo antifungal effect up to 82.4-100% disease suppression efficacy on greenhouse-grown pepper plants. The results obtained from this study may contribute to the development of new antifungal agents to protect the crops from fungal diseases.     

Screening potential bacterial biocontrol agents towards Phytophthora capsici in pepper

A total of 1,487 bacterial isolates were obtained from the rhizosphere, phyllosphere, endorhiza and endosphere of field-grown pepper. In a dual assay, 232 isolates displayed the antagonistic activity towards Phytophthora capsici L.; 36.6?% and 39.2?% of them were obtained from the rhizosphere and phyllosphere, respectively. 40 of the 232 antagonistic isolates producing inhibition zones of at least 5?mm in diameter were assessed for production of siderophores and chitinase, cellulose, and protease activity. These 40 isolates fell into 15 groups according to 90?% similarity of the banding patterns obtained by amplified ribosomal DNA restriction analysis (ARDRA). Seventeen isolates spanning the 15 groups were evaluated in greenhouse tests for their ability to control Phytophthora blight of pepper. Biocontrol efficacy ranged from 0.7?% to 92.3?%, with three isolates (B1301, R98, and PX35) exhibiting maximum ability to reduce the disease severity (83.5?%, 92.3?% and 83.5?%, respectively). Based on 16S rDNA sequencing, these isolates were identified as Bacillus cereus (B1301), Chryseobacterium sp (R98) and Bacillus cereus (PX35). This is the first report that Chryseobacterium sp. (R98) can function as a biocontrol agent of Phytophthora blight.     

Integrated pest management: A strategy to control resistance of Spodoptera exigua and Helicoverpa armigera caterpillars to insecticides on soybean in the Mekong Delta

The antibiotic nucleoside tubercidin produced by Streptomyces viola-ceoniger was evaluated for in-vivo efficacy and in-vitro activity against Phytophthora capsici, Magnaporthe grisea and Colletotrichum gloeosporioides. Tubercidin was more effective against P. capsici and M. grisea than against C. gloeosporioides in inhibiting mycelial growth. The bioassay on TLC plates was the most sensitive method and allowed the evaluation of antifungal activity of tubercidin even at a low concentration of 0.1 μgml^?1. As compared to the systemic fungicide metalaxyl, tubercidin was similar or somewhat higher in inhibition of mycelial growth of P. capsici. When applied to pepper stems, tubercidin was equally as effective as metalaxyl in the control of phytophthora blight in pepper plants, irrespective of application time and concentration. The treatment with 1000 μg ml^?1 tubercidin induced phytotoxicity in pepper plants. No control efficacy of phytophtora blight was observed in pepper plants supplied with a soil drench of tubercidin. Treatment with tubercidin at 500 μg ml^?1 completely protected pepper plants at first branch stage from phytophthora blight until four days after application. The control efficacy of tubercidin drastically declined seven days after application.     

Resistance of Phytophthora capsici to metalaxyl in plastic‐house capsicum crops in southern Italy*

In Calabria (southern Italy), control of crown and root rot of capsicum caused by Phytophthora capsici has relied primarily on soil drenches of metalaxyl. However, severe outbreaks occur every year in glasshouse crops, in which the practice of using plastic mulch and furrow irrigation favours the disease. Single‐hypha isolates of P. capsici collected in Calabria in 1992/1998 were tested in vitro for their level of sensitivity to metalaxyl. Isolates of other species of Phytophthora were used as reference. Fungicide sensitivity was determined by plating mycelial plugs onto potato dextrose agar amended with metalaxyl, at final concentrations ranging from 0.1 to 1000μg mL^?1 a.s. Inhibition of radial growth (%) was determined when colonies on unamended medium had covered approximately two‐thirds of the plate. The ED₅₀ values for inhibition of mycelial growth of P. capsici isolates ranged from 1.41 to44.6μg mL^?1 a.s. More than 80% of the P. capsici isolates from commercial plastic‐house crops in Calabria showed a moderate level of resistance as they were inhibited less than 60% at 5 μg mL^?1 but more than 60% at 100μg mL^?1     

Defence responses of pepper (Capsicum annuum L.) infected with incompatible and compatible strains of Phytophthora capsici

In this study, we investigated the activities of β-1,3-glucanase and peroxidase enzymes in the leaves of pepper cultivar A3 infected with the incompatible strain PC and the compatible strain HX-9 of Phytophthora capsici. The activities of β-1,3-glucanase and peroxidase enzymes substantially increased in the incompatible interactions compared to the compatible interactions. We also analysed the expression patterns of four defence-related genes, including CABPR1, CABGLU, CAPO1 and CaRGA1, in the leaves and roots of pepper inoculated with different strains of P. capsici. All gene expression levels were higher in the leaves than in the roots. Markedly different expression patterns were observed between incompatible and compatible host-pathogen interactions. In the incompatible interactions, the expression levels of CABPR1, CABGLU and CAPO1 genes in leaves increased by a maximum of 17.2-, 13.2- and 20.5-fold at 24, 12 and 12 h, respectively, whereas the CaRGA1 gene expression level increased to a lesser degree, 6.0-fold at 24 h. However, in the compatible interactions, the expression levels of the four defence-related genes increased by a maximum of 11.2-, 8.6-, 7.9- and 2.0-fold at 48, 24, 48 and 72 h, respectively. Compared to the leaves, the expression levels of the four defence-related genes were much lower in the roots. The highest levels of mRNA were those of the CABPR1 gene, which increased 5.1-fold at 24 h in the incompatible and 3.2-fold at 48 h in the compatible interactions. The other three genes exhibited lower expression levels in the incompatible and compatible interactions. These results further confirmed that defence-related genes might be involved in the defence response of pepper to P. capsici attack.     

拮抗真菌HTC的鉴定及其对辣椒疫病的生物防治潜力

为明确拮抗真菌HTC对辣椒疫霉Phytophthora capsici的拮抗机制,采用平板对峙、形态学鉴定和18S rDNA序列比对分析等方法对菌株HTC进行了鉴定,并研究其发酵液与抗生物质粗提液对辣椒疫霉不同发育阶段的影响。经鉴定,菌株HTC为金色毛壳菌Chaetomium aureum。在平皿对峙试验中,菌株HTC的红色分泌物能抑制辣椒疫霉菌丝的生长,抑制率为59.1%,后期HTC菌丝可缠绕并降解辣椒疫霉菌丝。发酵液与抗生物质粗提液对辣椒疫霉不同发育阶段均有抑制作用,发酵液对辣椒疫霉菌丝生长的抑制率高达97.58%,对辣椒疫病的防治效果均高于70%。浇灌发酵液后,可提高辣椒苗的苯丙氨酸解氨酶、过氧化物酶、多酚氧化酶的活性,并明显促进辣椒苗生长,鲜重和干重增加32.27%和18.09%。研究表明,金色毛壳菌菌株HTC是一株具有开发潜力的生防菌株。     

Control of dry bubble disease (Lecanicillium fungicola) in button mushroom (Agaricus bisporus) by spent mushroom substrate tea

Dry bubble (caused by Lecanicillium fungicola) is a widespread disease of button mushroom. The objective of the experiments was to determine the efficacy of compost teas made from spent mushroom substrate (SMS) as a biocontrol method against the disease. All SMS teas produced in this study significantly inhibited (100 %) the in vitro mycelial growth of L. fungicola, whereas the fungicide prochloraz at 50 ppm inhibited growth by 91 %. The in vivo effectiveness of two SMS aerated teas, one with mineral soil (MS) and the other with peat (TPT), was evaluated in two mushroom cropping trials inoculated with L. fungicola. The results demonstrated that the most effective treatments were those with TPT applied close to harvest and/or those with the greatest number of applications. The most efficacious treatments were TPT treatments (reducing disease by 34 to 73 % in the two trials, compared to the inoculated control). In contrast, prochloraz reduced disease by 7 % and 4 % in the two trials, compared to the control. These results suggest that dry bubble disease can be controlled by the use of spent mushroom substrate teas.     

Phytophthora species distribution in South African citrus production regions

Several Phytophthora spp. are known to cause a range of symptoms on citrus, resulting in significant crop losses worldwide. In South Africa, Phytophthora remains a destructive citrus disease, but the species and their distribution have not been well documented. A total of 162 Phytophthora isolates was collected from 60 citrus orchards in seven provinces of South Africa (Eastern Cape, Kwazulu-Natal, Limpopo, Mpumalanga, Northern Cape, North West and Western Cape). Isolates were identified to the species level through PCR-RFLP (restriction fragment length polymorphism) analyses of the internal transcribed spacer region. The identity of a subset of the isolates was confirmed using morphological and sequence analyses. Phytophthora nicotianae was the predominant species (76 % of isolates) and occurred in 80 % of the orchards in all of the provinces, followed by P. citrophthora (22 % of isolates in 28 % of orchards). The P. citrophthora isolates were further subdivided into two previously identified subgroups, G1 and G2, with most (69 %) of the isolates belonging to the G1 subgroup. Other Phytophthora species included P. multivora in the Western Cape Province, and an unknown species in the Eastern Cape Province with high sequence similarity (98 %) to a putative new species submitted to GenBank as Phytophthora taxon Sisuluriver. Phytophthora palmivora, a known citrus pathogen, was not identified. Most of the P. nicotianae isolates (79 %) were of the A1 mating type. The P. citrophthora isolates were mostly sterile (64 %), including most of the G1 isolates (81 %). The remaining G1 isolates (19 %) belonged to the A1 mating type, whereas almost all G2 isolates belonged to the A2 mating type except for one isolate that was sterile.     

An effective biocontrol bioformulation against Phytophthora blight of pepper using growth mixtures of combined chitinolytic bacteria under different field conditions

Phytophthora blight of pepper caused by Phytophthora capsici has devastating consequences when combined with other pathogens, including Rhizoctonia solani, Fusarium oxysporum, and Fusarium solani. In order to develop a field-effective biocontrol strategy against Phytophthora blight of pepper, three chitinolytic bacteria, Serratia plymuthica strain C-1, strongly antagonistic to P. capsici, Chromobacterium sp. strain C-61, strongly antagonistic to R. solani, and Lysobacter enzymogenes strain C-3, antagonistic to R. solani and Fusarium spp., were selected. In pot studies, application of cultures combining the three bacterial strains effectively suppressed Phytophthora blight more than application of any single bacterial strain. Bioformulations developed from growth of the strains in a simple medium containing chitin under large batch conditions resulted in effective control in field applications. Efficacy of the bioformulated product depended on both the dose and timing of application. Although the undiluted product suppressed Phytophthora blight under all field conditions, a 10-fold diluted product was effective in solar-sterilized greenhouses and in fields with crop rotation. These results suggest that the developed product could be a new effective system to control Phytophthora blight disease in pepper.     

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 %.     

Effect of aerated compost tea on grapevine powdery mildew, botrytis bunch rot and microbial abundance on leaves

Aerated compost tea (ACT), prepared from immature compost, was applied to foliage and fruit of grapevines (Vitis vinifera) to assess its potential for suppressing two important diseases: botrytis bunch rot, caused by Botrytis cinerea, and powdery mildew, caused by Erysiphe necator. An ACT applied to leaves of Cabernet Sauvignon vines in pots 7 days before inoculation with E. necator conidia reduced mean powdery mildew severity on the three youngest expanded leaves (at inoculation) to less than 1 %; mean severity on non-treated, inoculated leaves was 15 %. Multiple applications of ACTs at two vineyards in different growing seasons suppressed powdery mildew to <1 % mean severity on Chardonnay leaves (non-treated 79 % severity) and bunches (non-treated 77 % severity), and on Riesling leaves (non-treated 24 % severity). The same treatments reduced the incidence of Chardonnay bunches with latent B. cinerea and Riesling bunches with sporulating B. cinerea, although the level of botrytis bunch rot in both experiments was not economically damaging. The numbers of culturable bacteria, fungi and yeasts on Chardonnay leaves were higher than pre-treatment levels 10 days after ACT application, as were fungal numbers on Riesling leaves 21 days after treatment. Suppression by ACTs of two fruit and foliar pathogens of grapevine with different biology and epidemiology indicated potential for their use as a tactic in integrated disease management. Further testing of ACTs in a range of viticultural environments and application regimes will contribute to a better understanding of the impact of this tactic on disease, grape and wine quality.     

Development and validation of a standard area diagram set to assess blast severity on wheat leaves

This study aimed to develop and validate a standard area diagram set (SADS) to quantify the severity of blast, caused by Pyricularia oryzae, on wheat leaves. The SADs has ten levels: 0.1, 1, 5, 10, 22, 32, 42, 52, 62 and 72 % blast severity. To validate the SADs, 12 inexperienced raters estimated disease severity on 50 images of leaves from cultivars BR-18 (susceptible) and BRS-229 (partially resistant). Blast severity was first estimated without the use of the SADs on 50 leaves with a range of blast severity. The same raters evaluated the same 50 leaves using the SADs as an aid. The SADs improved accuracy (coefficient of bias, C _b ?=?0.88 and 0.99, without and with SADs, respectively) and agreement (Lin’s concordance correlation coefficient, ρ _c ?=?0.84 and 0.96 without and with SADs, respectively) of the estimates of severity. The absolute error was (-) 52 % without the SADs and (-) 24 % when using SADs as an aid. Severity estimates were more reliable when using SADs (R²?=?0.87 unaided and R²?=?0.92 with SAD). The SADs proposed in this study will improve accuracy and reliability of estimates of blast severity on wheat leaves.