Screening of plant extracts for organic management of downy mildew of sorghum

Antipathogenic potential of 38 plants was evaluated in the form of aqueous extracts against Peronoclerospora sorghi, causing downy mildew of sorghum. Conidial suspension and plant extracts were mixed individually and allowed to stand for 5 min and then used to inoculate the host by sprout-dip method. The sprouts thus inoculated were grown in pots, and the disease incidence was observed. Eight plant extracts (Cicer areatinum, Datura metel, Croton sparsiflorus, Parthenium hysterophorus, Nerium oleander, Chromolaena odorata, Duranta repens and Oxalis latifolia) at 20% concentration performed at par with chemical fungicide (Mancozeb 75%) by exhibiting total suppression of disease incidence to 0%, when compared with 64.1% of negative control. Organic management of air-borne inoculum of downy mildew of sorghum is feasible and preferable when compared with chemical control methods, considering human and environmental health concerns. The use of water extract keeps the technology simple so that it can be directly prepared and used by the farmers. Short-listing of eight most effective water extracts would help in self-reliance of farmers, reducing their dependence on commercial products.     

A novel technique for greenhouse evaluation of plant extracts for management of downy mildew of sorghum

Antipathogenic potential of 16 plants was evaluated in the form of aqueous extracts against Peronosclerospora sorghi, causing downy mildew of sorghum. A novel method was developed in which conidial suspension and plant extracts were mixed individually and allowed to stand for 5?min and then used to inoculate the host by sprout-dip method. The sprouts thus inoculated were grown in pots, and disease incidence was observed. Three extracts (Parthenium hysterophorus, Duranta repens and Oxalis latifolia) suppressed the disease at par with chemical fungicide (Mancozeb 75%). In form of spray, D. repens extract with tween-20 adjuvant (1.9% disease) performed closest behind mancozeb (0% disease), while negative control showed 21% disease incidence. Organic management of airborne inoculum of downy mildew of sorghum is feasible and preferable as compared to the use of chemical fungicides, considering human and environmental health concerns. Use of water extract keeps the technology simple so that it can be directly prepared and used by farmers. Short listing of three most effective water extracts would help in self-reliance of farmers, reducing their dependence on commercial products.     

Seed treatment with aqueous extract of Viscum album induces resistance to pearl millet downy mildew pathogen

Abstract

Downy mildew (Sclerospora graminicola [Sacc.] Schroet.) is a serious agricultural problem for pearl millet (Pennisetum glaucum [L.] R. Br.) grain production under field conditions. Six medicinally important plant species Azadirachta indica, Argemone mexicana, Commiphora caudata, Mentha piperita, Emblica officinalis and Viscum album were evaluated for their efficacy against pearl millet downy mildew. Seeds of pearl millet were treated with different concentrations of aqueous extract of the plants to examine their efficacy in controlling downy mildew. Among the plant extracts tested, V. album treatment was found to be more effective in enhancing seed quality parameters and also in inducing resistance against downy mildew disease. Germination and seedling vigor was improved in seeds treated with V. album extracts over control. Seeds treated with 10% concentration of V. album showed maximum protection against downy mildew disease under greenhouse and field conditions. The downy mildew disease protection varied from 44–70% with different concentrations. Leaf extract of V. album did not inhibit sporulation and zoospore release from sporangia of Sclerospora graminicola, indicating that the disease-controlling effect was attributed to induced resistance. Seed treatment with V. album extract increased pearl millet grain yield considerably. In V. album, treated pearl millet seedlings increased activities of peroxidase, and phenylalanine ammonia-lyase enzyme was detected. FTIR analysis of V. album extracts showed the presence of amides and other aromatic compounds which are antimicrobial compounds involved in plant defense.     

Evaluation of uncomposted botanical-based soil amendments for organic management of downy mildew of Sorghum

Uncomposted dried, powdered biomass of 38 selected botanicals was evaluated as soil amendments in a pot experiment for their potential to suppress downy mildew of sorghum caused by soil-borne oosporic inoculum of Perenosclerospora sorghi. A negative control showed 11.1% disease incidence. Six botanicals proved effective in form of dry biomass powder at 0.5% (w/w) amendment rate to control infection by oospores (Allium cepa 0%, Lantana camara 1.6%, Polyalthia longofolia 0%, Hyptis suaveolens 1.5%, Eucalyptus globulus 0% and Nerium oleander 1.6% of disease incidence). Dry powders are not only effective in disease suppression but confer additional benefits over composts like reduced bulk, increased shelf-life, consistent composition and results.     

The efficacy of spraying programmes and treatment methods against downy mildew in two grapevine cultivars in Kosovo

The present study aimed to assess the efficacy of applied programmes and treatment methods against downy mildew (Plasmopara viticola). Two vine cultivars (Frankovka and Game) were tested in an experimental field (in Rahovec, Kosovo) for a two-year period (2010–2011). The fungicides used were Quadris (azoxystrobin), Antracol EP-70 (propineb), Dithan M-45 (mancozeb), Ridomil (metalaxyl), Curzate (cymoxanil), Bordeaux mixture (calcium hydroxide and copper sulphate) and Mikal (fosetyl-aluminium) applied three treatment methods. The evaluation of the disease severity was performed using the McKinney index. In both tested cultivars and in both treatment years (2010 and 2011), all of the programmes tested significantly reduced the severity of grape downy mildew compared to the control. The highest efficacy against grape downy mildew was achieved with the combined use of Ridomil and Dithan (programme 3); this treatment was more than 80% effective in both cultivars and in both years.     

Comparative evaluation of composts and dried powdered biomass of botanicals for organic management of downy mildew of sorghum

The biomass of 10 selected botanicals was evaluated as soil amendments, in the form of composts and dry powder biomass, to compare their potential to suppress downy mildew disease caused by oosporic inoculum of Perenosclerospora sorghi. Eight botanicals (Allium cepa, Lantana camara, Polyalthia longifolia, Hyptis suaveolens, Azadirachta indica, Nerium oleander Eucalyptus globulus and Tamarindus indica) proved effectively in the form of dry biomass powder at 4% (w/w) rate to control infection by oospores, while two (Croton sparsiflorus and Tagetes erecta) promoted it. In the form of composts (2% rate), the efficacy of botanicals was generally found reduced with the appearance of more disease than negative control in case of all botanicals except one (A. cepa). At the 4% compost rate, only four botanicals namely, A. cepa, P. longifolia, H. suaveolens and L. camara were found to suppress the disease. At higher amendment rates of 6, 8 and 10%, six botanicals suppressed the disease. Dry powders are not only superior in disease suppression but confer additional benefits like reduced bulk, increased shelf life, consistent composition and results.     

Genetic structure of Italian population of the grapevine downy mildew agent,Plasmopara viticola

Downy mildew, caused by the Oomycete Plasmopara viticola, is one of the most important diseases affecting the Eurasian grapevine, Vitis vinifera. The pathogen originated in Northern America and its presence was signalled for the first time into Europe in 1878. In this study, the genetic variability and structure of Italian P. viticola population was investigated by genotyping 106 P. viticola strains belonging to 12 different regions, at 31 microsatellite markers. As a result of the high percentage of missing data, 96 strains and 19 loci were retained for the data analysis. The overall Italian population presents absence of clones, evidence of sexual and asexual reproduction and a low genetic diversity, as expected for an introduced pathogen, but a slightly higher genetic diversity than that found in other European populations, based on allelic diversity at the investigated microsatellite loci. Out of 19 loci, half shows deviation from Hardy–Weinberg equilibrium and, indeed, structure analysis indicates the presence of two separate genetic clusters, with little but significantly different distribution according to geography (west–east gradient) and climatic conditions. Overall, the analysis of the P. viticola population, 140 years after its appearance in Italy, provides indication on the pathogen adaptability. This should be taken into consideration in the future for preserving the durability of disease resistant varieties in open field. In this view, all the disease control methods available should be integrated in order to reduce the selection of pathogen strains able to overcome plant resistance.     

黄瓜霜霉病及寄主抗性机制研究进展

在黄瓜生产中,由古巴假霜霉菌(Pseudoperonospora cubensis)引起的霜霉病危害严重,影响叶、茎和花序生长发育,导致黄瓜产量及品质降低。通过对黄瓜霜霉病的病原菌检测和防御途径、影响及调控因素、抗病原菌候选基因发掘、蛋白质组和基因组分析、黄瓜霜霉病QTL连锁标记开发及其抗病育种等多方面的最新进展进行综述,以期为今后进一步揭示黄瓜乃至农作物对霜霉病的抗性机制研究提供借鉴和参考。     

Microsatellite DNA markers for Plasmopara viticola,the causal agent of downy mildew of grapes

Microsatellite loci were isolated from Plasmopara viticola (Oomycetes), the causal agent of downy mildew of grape, one of the most damaging fungal diseases of grapevine worldwide. Seven polymorphic loci were obtained from an enriched partial genomic library. A low genetic diversity was observed at all loci, with a mean observed allele number of 3.75 and an observed heterozygosity ranging from 0.074 to 0.547. Cross‐amplification tests on three closely related taxa indicated that two loci could be used in other Oomycetes species. These microsatellite loci were proved to be useful for population genetic analysis.     

Characterisation of pathogenic and molecular diversity in Sclerospora graminicola,the causal agent of pearl millet downy mildew

Genotypic diversity among 46 isolates of Sclerospora graminicola collected from seven states in India during 1992–2005 was determined through pathotyping and AFLP analysis. A high level of variation was observed among the isolates for downy mildew incidence, latent period and virulence index. Based on the reaction on a set of nine pearl millet lines, 46 isolates were classified in 21 pathotypes. Quantitative differences in virulence levels of the test isolates were assessed by calculating the virulence index (disease incidence × latent period ^? 1). A dendrogram generated by the average linkage cluster analysis of virulence index clustered the 46 isolates into eight groups. Region-specific grouping of five isolates from Gujarat and six from Rajasthan was observed within two distinct groups. Temporal variation was also observed among the isolates collected from the same location and same host over the years. A total of 297 bands were scored following selective amplification with three primer combinations E-TT/M-CAG, E-AT/M-CAG and E-TG/M-CAT and all of them were polymorphic. Cluster analysis of AFLP data clustered the test isolates into seven groups. Analysis of molecular variance indicated that variation in the S. graminicola populations was largely due to differences among the isolates within the states.     

Effect of combination between inducer resistance chemicals and systemic fungicides on management of sweet melon downy mildew

The efficiency of some inducer resistance chemicals (IRCs) like bion, chitosan, humic acid and salicylic acid as well as the fungicides like Folu-Gold, Galben Copper, Previcure-N and Redomil Gold Mancozeb on management of sweet melon downy mildew, caused by Pseudoperonospora cubensis was evaluated in vitro and in vivo. Also, the efficiency of the alternation between the sprayed two fungicides and IRCs on management of the disease and the produced fruit yield and its total soluble solids (TSS) under field conditions were assessed. The inhibitory effect of the IRCs and the tested fungicides on sporangial germination of P. cubensis resulted in a significant reduction in the germinated sporangia. In addition, IRCs were less effective than the fungicides. Disease management revealed the same trend of the in vitro experiment when they sprayed fungicides on sweet melon plants artificially inoculated with the sporangia of the causal fungus under greenhouse conditions. Furthermore, under field conditions, spraying sweet melon plants with the two tested fungicides was the most efficient in decreasing the disease and increasing fruit yield and its TSS, to somewhat, followed by the alternation between them and the tested IRCs. In addition, IRCs treatments showed the lowest efficiency in this regard.     

Enhancement of downy mildew of sorghum by some organic soil amendments: a caution to organic farmers

Matured botanical compost of Croton sparsiflorus and Azadirachta indica were evaluated for promotion of sorghum downy mildew at the rate of 0.5% (w/w). Exceptionally high promotional rates of 26.9% (C. sparsiflorus) and 30.1% (A. indica) were observed as compared to negative control (8.8%). The enhancement of disease was higher than that obtained by the supply of recommended dose of urea (13.3%). As the pathogen (Peronosclerospora sorghi) is a biotroph and cannot multiply by using organics as nutrient source in the absence of a host, the mechanism of disease enhancement is inferred to be through stimulation of oospore germination by organic compounds.     

Cerebroside as an elicitor for induced resistance against the downy mildew pathogen in pearl millet

Innate defence mechanisms in plants can be triggered and enhanced by certain agents, which are referred to as inducers. Inducing resistance against a broad spectrum of pathogens in otherwise susceptible plants is seen as a potentially safer alternative to other methods of chemical control of plant diseases. Cerebrosides, which are glycosphingolipids extracted from various plant pathogens, have been reported as resistance elicitors in the rice‐pathogen system. In the present study, cerebroside elicited resistance against downy mildew disease (caused by Sclerospora graminicola) of pearl millet (Pennisetum glaucum) that was highly significant. The resistance was of systemic nature and the time required for the resistance to build up was from 2 days onwards. There was a significant yield enhancement due to disease suppression by cerebroside treatment. Promising results were obtained in a preliminary field trial.     

Resurgence of cucurbit downy mildew in the United States: Insights from comparative genomic analysis of Pseudoperonospora cubensis

Pseudoperonospora cubensis, the causal agent of cucurbit downy mildew (CDM), is known to exhibit host specialization. The virulence of different isolates of the pathogen can be classified into pathotypes based on their compatibility with a differential set composed of specific cucurbit host types. However, the genetic basis of host specialization within P. cubensis is not yet known. Total genomic DNA extracted from nine isolates of P. cubensis collected from 2008 to 2013 from diverse cucurbit host types (Cucumis sativus, C. melo var. reticulatus, Cucurbita maxima, C. moschata, C. pepo, and Citrullus lanatus) in the United States were subjected to whole‐genome sequencing. Comparative analysis of these nine genomes confirmed the presence of two distinct evolutionary lineages (lineages I and II) of P. cubensis. Many fixed polymorphisms separated lineage I comprising isolates from Cucurbita pepo, C. moschata, and Citrullus lanatus from lineage II comprising isolates from Cucumis spp. and Cucurbita maxima. Phenotypic characterization showed that lineage II isolates were of the A1 mating type and belonged to pathotypes 1 and 3 that were not known to be present in the United States prior to the resurgence of CDM in 2004. The association of lineage II isolates with the new pathotypes and a lack of genetic diversity among these isolates suggest that lineage II of P. cubensis is associated with the resurgence of CDM on cucumber in the United States.     

Phenol metabolism in downy mildew resistant and susceptible genotypes of pearl millet

Biochemical characterisation of pearl millet genotypes was carried at pre- (45 DAS) and post-infection (57 DAS i.e. 7 days after infection) stages. Total phenol content at pre-infection stage did not show inherent resistance or susceptibility. While the total phenol content was found to be higher in susceptible genotypes at post-infection stage, qualitative analysis of phenol through high-performance thin layer chromatography showed absence of ferulic acid in resistant genotypes at pre-infection stage. Peroxidase (POX) activity was higher in susceptible genotypes at both the stages of analysis. Constitutive activity of phenylalanine ammonia lyase was higher in resistant genotype whereas induced activity was recorded higher in susceptible genotypes. Native poly-acrylamide gel electrophoresis isozyme banding pattern of POX showed some inducible band(s) due to disease infection in resistant and susceptible genotypes.     

Sclerospora graminicola- and arachidonic acid-induced autofluorescence in downy mildew resistant and susceptible genotypes of pearl millet

Autofluorescence of downy mildew resistant and susceptible cells of pearl millet seedlings undergoing hypersensitive reaction (HR) upon Sclerospora graminicola-inoculation and arachidonic acid (AA)-treatment was studied. Two-day-old seedlings of a highly resistant (IP 18296) and a highly susceptible (23D₂B) genotype of pearl millet were either inoculated with zoospore suspension of S. graminicola or treated with AA for 24 h. The coleoptiles with hypersensitive necrotic spots were processed by the standard procedure, and the tissues were subjected to fluorescence microscopy. A differential accumulation of autofluor-escent compounds in resistant and susceptible pearl millet genotypes was observed with most accumulation occurring in resistant cells treated with AA. The variation in the degree of fluorescence and the spatial accumulation of autofluorescent compounds among the two inoculated/treated genotypes is discussed.     

Haplotyping and mapping a large cluster of downy mildew resistance gene candidates in sunflower using multilocus intron fragment length polymorphisms

Downy mildew (Plasmopara halstedii (Farl.) Berlese et de Toni) is a serious foliar pathogen of cultivated sunflower (Helianthus annuus L.). Genetic resistance is conditioned by several linked downy mildew resistance gene specificities in the HaRGC1 cluster of TIR-NBS-LRR resistance gene candidates (RGCs) on linkage group 8. The complexity and diversity of the HaRGC1 cluster was assessed by multilocus intron fragment length polymorphism (IFLP) genotyping using a single pair of primers flanking a hypervariable intron located between the TIR and NBS domains. Two to 23 bands were amplified per germplasm accession. The size of the included intron ranged from 89 to 858 nucleotides. Forty-eight unique markers were distinguished among 24 elite inbred lines, six partially isogenic inbred lines, nine open-pollinated populations, four Native American land races, and 20 wild H. annuus populations. Nine haplotypes (based on 24 RGCs) were identified among elite inbred lines and were correlated with known downy mildew resistance specificities. Sixteen out of 39 RGCs identified in wild H. annuus populations were not observed in elite germplasm. Five partially isogenic downy mildew resistant lines developed from wild H. annuus and H. praecox donors carried eight RGCs not found in other elite inbred lines. Twenty-four HaRGC1 loci were mapped to a 2-4 cM segment of linkage group 8. The multilocus IFLP marker and duplicated, hypervariable microsatellite markers tightly linked to the HaRGC1 cluster are powerful tools for distinguishing downy mildew resistance gene specificities and identifying and introgressing new downy mildew resistance gene specificities from wild sunflowers.     

Pearl millet downy mildew as affected by seed dressing and cultivar in the Sahel and Sudan savanna of Nigeria

Field trials in which seed dressing with Apron Star 42 WS (metalaxyl-M) was tested on five pearl millet cultivars were laid at Gashua (Sahel) and Maiduguri (Sudan Savanna) for three wet seasons (1998–2000). Using the split-plot design, seed dressing and cultivars were tested in main- and sub-plots, respectively. Results showed that at each location and season, there was significantly high seedling emergence and grain yield but lower downy mildew incidence as a result of seed dressing with metalaxyl-M compared with check. Among the cultivars tested, SOSAT C-88 had significantly lower downy mildew incidence and the highest grain yield during the three seasons. Dressed with metalaxyl-M, other cultivars had lower downy mildew and higher grain yield than undressed check. It is therefore important that pearl millet should be dressed with metalaxyl as routine practice before cultivation in the arid area of Nigeria.     

Comparative evaluation of Pseudomonas fluorescens and their lipopolysaccharides as implicated in induction of resistance against pearl millet downy mildew

Two plant growth promoting Pseudomonas fluorescens isolates namely UOM SAR 14 and UOM SAR 80 most effectively induced resistance against downy mildew disease of pearl millet both under greenhouse and field conditions. Relative assessment of live cultures of P. fluorescens UOM SAR 14 and UOM SAR 80 and their lipopolysaccharides (LPS) extracted from their cell walls were evaluated for their ability to induce resistance against pearl millet downy mildew. Treatment with P. fluorescens and their LPS enhanced the seed germination and seedling vigour considerably. Although both live cultures and their LPS treatment induced resistance in pearl millet against downy mildew disease both under greenhouse and field conditions as evidenced by the significant reduction of the disease, live cultures were more effective than the LPS in level of resistance induced. Live cultures of UOM SAR 14 and UOM SAR 80 induced 66% and 57% protection while their respective LPS extracts offered 59 and 53% protection against downy mildew disease under greenhouse conditions. Similarly, under field conditions with very heavy inoculum pressure live cultures offered 75% and 70%, and their LPS offered 71% and 67% protection, respectively. In either case, the time gap required for the building up of resistance was found to be 3 days and nature of the resistance induced was systemic and durable with both live cultures and their lipopolysaccharides. It was also noticed that the live bacteria significantly varied in the degree of protection offered and so also their respective LPS.