Downy mildew resistance evaluation in 28 grapevine hybrids promising for breeding programs in Trentino region (Italy)

Downy mildew is a major grapevine disease caused by the biotrophic oomycete, Plasmopara viticola. Numerous disease resistance studies of diverse Vitis germplasm have been previously carried out to identify downy mildew resistance sources; however, ratings were mainly reported using leaf disc in vitro testing and foliage field assessment, or upon leaf and cluster field evaluations. In the current study, 28 grapevine hybrid cultivars were screened using leaf disc bioassay, for disease resistance characterization of both existing and wild-collected materials. 16 hybrids were identified as highly resistant or resistant, and will serve as relevant resistance donors in future pre-breeding and breeding programs. All grapevine hybrids were evaluated for foliar and cluster downy mildew resistance in an untreated field trial over three successive years. This study showed that the leaf disc bioassay provided some information on the resistance level of the genotypes under scrutiny, but it was a weak predictor of their resistance level under field conditions on leaves and even more on bunches. These findings are relevant to future applications in both traditional and marker-assisted breeding programs which promote sustainable viticulture.     

Evaluation and QTL mapping of resistance to powdery mildew in lettuce

Lettuce (Lactuca sativa) is the major leafy vegetable that is susceptible to powdery mildew disease under greenhouse and field conditions. Quantitative trait loci (QTLs) for resistance to powdery mildew under greenhouse conditions were mapped in an interspecific population derived from a cross between susceptible L. sativa cultivar Salinas and the highly susceptible L. serriola accession UC96US23. Four significant QTLs were detected on linkage groups LG 1 (pm‐1.1), LG 2 (pm‐2.1 and pm‐2.2) and LG 7 (pm‐7.1), each explaining between 35 to 42% of the phenotypic variation. The four QTLs are not located in the documented hotspots of lettuce resistance genes. Alleles for the disease resistance at the four QTLs originated from both parents (two from each), demonstrating that even highly susceptible accessions may provide alleles for resistance to powdery mildew. These QTLs appeared to operate during limited periods of time. Results of the field trials with F_2:3 and F_3:4 families derived from a Soraya (moderately resistant) × Salinas cross demonstrated effective transfer of resistance to powdery mildew in this material. An integrated rating approach was used to estimate relative levels of resistance in 80 cultivars and accessions tested in a total of 23 field and greenhouse experiments. Generally, very low resistance was observed in crisphead‐type lettuces, while the highest relative resistance was recorded in leaf and butterhead types. Comparison of two disease assessment methods (percentage rating and the area under the disease progress steps, AUDPS) for detection of QTLs shows that the two approaches complement each other.     

Characterization of resistance mechanisms activated by Trichoderma harzianum T39 and benzothiadiazole to downy mildew in different grapevine cultivars

Downy mildew, caused by Plasmopara viticola, is one of the most destructive diseases of grapevine and is controlled with intense application of chemical fungicides. Treatment with Trichoderma harzianum T39 (T39) or benzothiadiazole‐7‐carbothioic acid S‐methyl ester (BTH) has been previously shown to activate grapevine resistance to downy mildew and reduce disease symptoms in the Pinot noir cultivar. However, enhancement of plant resistance can be affected by several factors, including plant genotype. In order to further extend the use of resistance inducers against downy mildew, the physiological and molecular properties of T39‐ and BTH‐activated resistance in different cultivars of table and wine grapes were characterized under greenhouse conditions. T39 treatment reduced downy mildew symptoms, but the degree of efficacy differed significantly among grapevine cultivars. However, efficacy of BTH‐activated resistance was consistently high in the different cultivars. Expression profiles of defence‐related genes differed among cultivars in response to resistance inducers and to pathogen inoculation. T39 treatment enhanced the expression of defence‐related genes in the responsive cultivars, before and after P. viticola inoculation. A positive correlation between the efficacy of T39 and the expression level of defence‐related genes was found in Primitivo and Pinot noir plants, while different genes or more complex processes were probably activated in Sugraone and Negroamaro. The data reported here suggest that the use of a responsive cultivar is particularly important to maximize the efficacy of resistance inducers and new natural inducers should be explored for the less responsive cultivars.     

Investigations on the role of cuticular wax in resistance to powdery mildew in grapevine

Cuticular wax on the plant epidermis inhibits or enhances prepenetration events of powdery mildew (Erysiphe necator Schwein). We examined the role of cuticular leaf and berry waxes as a resistance mechanism in four grapevine genotypes (Italia?×?Mercan-174, Gürcü, Isabella, Özer Karas?) resistant to powdery mildew after natural infection and inoculation. To understand cuticular wax properties, we determined the amount of wax and antifungal effects of thin layer chromatography (TLC) fractions from cuticular leaf and berry waxes, then assessed the chemical composition of fractions with different antifungal activities using gas chromatography/mass spectrometry (GC/MS). Susceptible genotypes Cabernet Sauvignon and Italia were used for comparison. Resistant and sensitive genotypes did not differ significantly in the total amount of wax on leaves and berries; however, 27 fatty acids, 26 alkanes, 6 terpenes, 4 indole derivatives and 4 ketones, and 3 amides, 3 phenols and 3 steroids were detected in fractions with high antifungal activity (≥75% inhibition of germination) in leaf and/or berry cuticular waxes of resistant genotypes only. These compounds may be evaluated as markers for powdery mildew resistance during genotype selection in a grapevine breeding program.     

A standardised method for the quantitative analysis of resistance to grapevine powdery mildew

Powdery mildew caused by Erysiphe necator is one of the most important diseases affecting grapevine (Vitis vinifera, L.). Control of this pathogen is based on the use of fungicides, which cause environmental damage and increase production costs. A cost-effective and environmentally friendly alternative to control the disease relies on using resistant varieties. While most V. vinifera cultivars are susceptible to powdery mildew, several species belonging to the Vitaceae have been described as resistant. Several loci for resistance to grapevine powdery mildew have been identified through genetic analysis of segregating populations derived from different resistance sources. Identifying quantitative trait loci (QTL) with minor effects on the resistance may prove valuable in a strategy of pyramiding, which aims at increasing the durability of the resistance. However, current methods for evaluation of resistance either do not take into account quantitative variations, or, if they do, are not adapted to large sample sets. Here we develop a method for the analysis of quantitative resistance to grapevine powdery mildew in large populations. We devised a semi-quantitative resistance scale and confirmed the usefulness of a cell counter to quantify sporulation. We compared three inoculation methods and identified dry inoculation using a settling tower as the one giving the best infection. Finally, we confirmed the value of the method by applying it to a set of plants segregating for resistance to E. necator. Using the method described here for the quantitative analysis of the resistance to powdery mildew will prove valuable for breeding for durable resistance.     

83份西农系小麦品种（系）抗性鉴定及抗病基因分子检测

为西北农林科技大学小麦新育成品种（系）在黄淮麦区的大面积推广,该研究对83份西农新育成的小麦品种（系）进行苗期抗条锈病和白粉病鉴定,成株期抗条锈病、白粉病、叶锈病和赤霉病鉴定,并在田间自然环境下对其抗性进行鉴定及对相关抗病基因进行分子检测。结果显示,在苗期人工接种鉴定中,有63、29和16份小麦品种（系）分别对条锈菌Puccinia striiformis f.sp.tritici生理小种CYR32、CYR33和CYR34表现出抗性,9份小麦品种（系）对3个条锈菌生理小种均表现出抗性;有10、3和0份小麦品种（系）分别对白粉菌Blumeria graminis f.sp.tritici生理小种E15、E09和A13表现出抗性。在成株期人工接种鉴定中,有23、15、28和62份小麦品种（系）分别对条锈病、白粉病、叶锈病和赤霉病表现出抗性。在83份小麦品种（系）中有6份在苗期和成株期均对小麦条锈病表现出抗性。在田间抗性鉴定中,有57、6、65和40份小麦品种（系）分别对条锈病、白粉病、赤霉病及叶锈病表现出抗性。在83份小麦品种（系）中,3份含有Yr5基因,22份含有Yr9基因,3份含有Yr17基因,2份含有Pm24基因,14份含有Lr1基因,所占比例分别为3.6%、26.5%、3.6%、2.4%和16.8%。     

Variation in adult plant resistance to powdery mildew in spring oats under field and laboratory conditions

Ten oat genotypes were assessed for adult plant resistance (APR) to powdery mildew ( Erysiphe graminis f.sp. avenae ) in small field plots and on detached leaf segments in the laboratory using glasshouse-grown plants. Significant genotype differences were observed in the field and on leaf segments for latent period, infection frequency and spore production, components used to assess APR. The genotype OM1711, which was selected because it expressed a higher level of mildew resistance than its most resistant parent, cv. Maldwyn, showed this transgressive resistance in both field and laboratory tests. By contrast, the genotype OM1387, also selected for high mildew resistance, only showed this enhancement in the field. It is concluded that quantitative assessment of mildew in detached leaves is not sufficiently precise to be used alone in selecting segregants for high levels of APR, and that field data are also needed.     

Activity of the New Fungicide Benthiavalicarb Against Plasmopara viticola and its Efficacy in Controlling Downy Mildew in Grapevines

Benthiavalicarb is a new fungicide active against Oomycetes fungal plant pathogens. The present study shows that benthiavalicarb is effective for controlling the Oomycete fungal pathogen Plasmopara viticola, which causes downy mildew in grapevines. The fungicide did not affect zoospore discharge from sporangia of P. viticola, but strongly inhibited zoospore encystment, cystospore germination in vitro and mycelial growth, together with sporangial production in vivo. Benthiavalicarb showed strong prophylactic and local activity in intact plants or detached leaves and low translaminar activity. The compound was not translocated from leaf to leaf in either a acropetal or basipetal direction. Benthiavalicarb applied at 1, 3 and 6 days post-inoculation protected grapevine plants against downy mildew and inhibited sporulation of the pathogen. Similar results were obtained on leaf disks if benthiavalicarb was applied up to 96 h post-inoculation. Benthiavalicarb diminished the sporulation of P. viticola when applied to established disease in the tissue. Benthiavalicarb remained active on leaves for a period up to 28 days. Two foliar applications of benthiavalicarb, 2 weeks apart, to field-grown grapevines inhibited downy mildew development and were as effective as the standard metalaxyl-Cu treatment in controlling the disease. A formulated mixture of benthiavalicarb + Folpet was similar or superior in performance to metalaxyl-Cu and the new strobilurin trifloxystrobin in controlling downy mildew. The effectiveness of benthiavalicarb makes it well suited for integration into a control programme against downy mildew disease in vineyards, and as a component to delay resistance buildup.     

Differential expression of resistance to powdery mildew incited by race 1 or 2 ofsphaerotheca fuliginea inCucumis melo genotypes at various stages of plant development

Thirty-nine genotypes ofCucumis melo (plant introduction entries, open-pollinated cultivars and F₁ hybrids) were evaluated for resistance to powdery mildew under either natural field conditions or artificial inoculation in growth chambers at the cotyledonary stage and the 2-true-leaf stage. Results confirmed that susceptibility in cotyledons was not necessarily associated with susceptibility in either true leaves in growth chambers or adult plants in the field. However, resistance at the 2-true-leaf stage in growth chambers was highly correlated with resistance of field-grown plants. Results also showed that 20 muskmelon genotypes resistant to race 1 at the cotyledonary stage were also resistant at the 2-leaf-stage and as adult plants in the field. The same was true for ten genotypes with race 2 inoculations. Because muskmelon genotypes expressing resistance in cotyledons were also resistant in true leaves in growth chambers or the field, the use of plants at the cotyledonary stage is recommended for screening for powdery mildew resistance caused by race 1 or race 2 ofS. fuliginea. When cotyledons are susceptible, screening should be done at the 2-true-leaf stage.     

Coincidence of maximum severity of powdery mildew on grape leaves and the carbohydrate sink‐to‐source transition

Grapevine leaves infected with powdery mildew are a source of inoculum for fruit infection. Leaves emerging on a single primary shoot of Vitis vinifera cv. Cabernet Sauvignon were exposed to average glasshouse temperatures of 18°C (0·23 leaves emerging/day) or 25°C (0·54 leaves emerging/day). All leaves on 8–10 shoots with approximately 20 leaves each were inoculated with Erysiphe necator conidia to assess disease severity after 14 days in the 25°C glasshouse. Two photosynthetic ‘source’ leaves per shoot on the remaining 8–10 shoots were treated with ¹⁴CO₂ to identify, by autoradiography, the leaf position completing the carbohydrate sink‐to‐source transition. There was a clear association between the mean modal leaf position for maximum severity of powdery mildew (position 3·7 for 18°C; position 4·4 for 25°C) and the mean position of the leaf completing the sink‐to‐source transition (position 3·8 for 18°C; position 4·7 for 25°C). The mean modal leaf position for the maximum percentage of conidia germinating to form secondary hyphae was 4·2 for additional plants grown in the 25°C glasshouse. A higher rate of leaf emergence resulted in a greater proportion of diseased leaves per shoot. A Bayesian model, consisting of component models for disease severity and leaf ontogenic resistance, had parameters representing the rate and magnitude of pathogen colonization that differed for shoots developing in different preinoculation environments. The results support the hypothesis that the population of leaves in a vineyard capable of supporting substantial pathogen colonization will vary according to conditions for shoot development.     

Induced resistance to foliar diseases by soil solarization and Trichoderma harzianum

The effect of soil solarization and Trichoderma harzianum on induced resistance to grey mould (Botrytis cinerea) and powdery mildew (Podosphaera xanthii) was studied. Plants were grown in soils pretreated by solarization, T. harzianum T39 amendment or both, and then their leaves were inoculated with the pathogens. There was a significant reduction in grey mould in cucumber, strawberry, bean and tomato, and of powdery mildew in cucumber, with a stronger reduction when treatments were combined. Bacillus, pseudomonad and actinobacterial communities in the strawberry rhizosphere were affected by the treatments, as revealed by denaturing gradient gel electrophoresis fingerprinting. In tomato, treatments affected the expression of salicylic acid (SA)‐, ethylene (ET)‐ and jasmonic acid (JA)‐responsive genes. With both soil treatments, genes related to SA and ET – PR1a, GluB, CHI9 and Erf1 – were downregulated whereas the JA marker PI2 was upregulated. Following soil treatments and B. cinerea infection, SA‐, ET‐, and JA‐related genes were globally upregulated, except for the LOX genes which were downregulated. Upregulation of the PR genes PR1a, GluB and CHI9 in plants grown in solarized soil revealed a priming effect of this treatment on these genes' expression. The present study demonstrates the capacity of solarization and T. harzianum to systemically induce resistance to foliar diseases in various plants. This may be due to either a direct effect on the plant or an indirect one, via stimulation of beneficial microorganisms in the rhizosphere.     

Field evaluation of mungbeans for resistance to Cercospora leaf spot and powdery mildew

Abstract

Cercospora leaf spot, caused by Cercospora canescens and powdery mildew, caused by Erysiphe polygoni are two of the most important fungal diseases of mungbeans. Nearly 4000 accessions of the global mungbean collection at the Asian Vegetable Research and Development Center were screened for resistance to these two pathogens. Less than 4% and 12% of the accessions showed resistance to Cercospora leaf spot and powdery mildew, respectively. The level of resistance to Cercospora leaf spot varied and highly resistant lines were not found. A few lines were rated highly resistant to powdery mildew over several years but others were rated moderately resistant or susceptible in other years.     

不同栽培模式下葡萄霜霉病菌遗传结构的时间动态分析

为明确不同栽培模式下葡萄霜霉病菌Plasmopara viticola的遗传结构、遗传多样性及遗传分化水平,于2014-2015年定期采集露地和避雨2种栽培模式下的葡萄霜霉病菌菌株,利用6对SSR引物对该病菌基因型、遗传多样性及遗传分化进行对比分析。结果表明,露地和避雨栽培模式下葡萄霜霉病菌群体的Nei’s基因多样性指数大于0.14,香农多样性指数大于0.31,2种栽培模式下群体具有丰富的遗传多样性,但避雨栽培模式可显著降低群体等位基因数和等位基因频率。露地栽培模式下该病菌群体的流行模式呈现中等水平无性繁殖,2年初侵染和再侵染对病害流行的贡献率分别约占26.1%和73.9%;避雨栽培模式下葡萄霜霉病菌群体的流行模式则呈现高等水平无性繁殖,初侵染和再侵染对病害流行的贡献率分别约占4.3%和95.7%。卵孢子的形成对于葡萄霜霉病菌种群遗传变异和有效越冬起着关键的作用。2014-2015年露地栽培模式下葡萄霜霉病菌群体的主效流行基因型对病害流行的贡献率分别为44.5%和51.8%;而其在避雨栽培模式下葡萄霜霉病菌群体的贡献率分别可达84.2%和87.1%。同一年份的露地和避雨栽培模式下葡萄霜霉病菌群体的主效基因型种类相同,2个群体间的等位基因频率呈现显著正相关性,且二者之间存在频繁的基因交流,推测避雨栽培模式下葡萄霜霉病的初侵染源自于避雨设施附近的露地栽培病株上再侵染形成的飞散传播孢子囊。     

Crop damage caused by powdery mildew on hop and its relationship to late season management

Powdery mildew of hop (Podosphaera macularis) may cause economic loss due to reductions in cone yield and quality. Quantitative estimates of crop damage from powdery mildew remain poorly characterized, especially the effect of late season disease management on crop yield and quality. Field studies in Washington State evaluated cone yield, bittering acid content and quality factors when fungicide applications were ceased at different stages of cone development. The incidence of cones with powdery mildew was linearly correlated with yield of cones, bittering acids and accelerated cone maturation. In cultivar Galena, the cumulative effect of every 1% increase in cones powdery mildew incidence was to reduce alpha‐acid yield by 0·33%, which was due to direct effects on cone yield but also indirect effects mediated by dry matter. In the more susceptible cultivar Zeus, alpha‐acid yield was increased 20% by controlling powdery mildew through the transition of bloom to early cone development compared to ceasing fungicide applications at bloom: additional applications provided only modest improvements in alpha‐acid yield. In both cultivars, the impact of powdery mildew on aroma characteristics and bittering acid content were less substantial than cone yield. The damage caused by powdery mildew to cone colour and alpha‐acid yield, as well as the effectiveness of fungicide applications made to manage the disease, appears inseparably linked to dry matter content of cones at harvest. Realising achievable yield potential in these cultivars requires control of the disease through early stages of cone development and harvest before maturity exceeds c. 25% dry matter.     

Macroscopic and Histological Characterisation of Genes er1 and er2 for Powdery Mildew Resistance in Pea

In pea, two single recessive genes, er1 and er2, have been identified for resistance to powdery mildew caused by Erysiphe pisi, but little is known about their mode of action. Pea accessions carrying the genes er1 or er2 and other accessions displaying resistance to powdery mildew in the field were studied. In accessions carrying gene er1, epidermal cell penetration was prevented and very few haustoria or colonies were formed. Under controlled conditions, er1 conferred complete or almost complete resistance to the fungal isolates used and this resistance was not associated with macroscopically visible necrosis. Under field conditions these accessions developed a low level of disease. Resistance in line JI2480 (carrying er2) increased with temperature and leaf age, and complete resistance was expressed only at high temperature (25 °C) or in mature leaves. This resistance was based mainly on post-penetration cell death, complemented by a reduction of percentage penetration success in mature leaves. Combining the resistance provided by gene er1 and by line JI2480 into new cultivars is likely to increase their level of resistance and enhance durability of the protection.     

An Aqueous Extract of the Dry Mycelium of Penicillium chrysogenum Induces Resistance in Several Crops under Controlled and Field Conditions

We have examined the effect of Pen, an aqueous extract of the dry mycelium of Penicillium chrysogenum, on plant–pathogen interactions. Pen controlled a broad range of pathogens on several crop plants under greenhouse and field conditions. Pen protected grapevine from downy and powdery mildew (caused by Plasmopara viticola and Uncinula necator), tomato from early blight (caused by Phytophthora infestans), onion from downy mildew (Peronospora destructor) and apple trees from apple scab (caused by Venturia inaequalis) to a similar extent as fungicides such as copper and sulphur or well-known inducers such as benzothiadiazole or β-aminobutyric acid. Pen had no major direct fungicidal effect and is thus supposed to protect plants by activating their defense mechanisms. The raw material for extraction of Pen was available in constant quality, a prerequisite for commercial application. Under certain conditions, Pen caused phytotoxic side effects. The symptoms mostly consisted of small necrotic spots or, more rarely, of larger necrotic areas. The development of the symptoms was dependent on several parameters, including concentration of Pen, the number of applications, the persistence on the plant tissue, the plant species and variety and environmental conditions. In grapevine, a partially purified fraction of Pen was much less toxic than the crude Pen extract, but protected the plants to a similar extent against P. viticola. Our data show that Pen has interesting and unique properties as a plant protection agent, but more research is needed to further reduce its phytotoxic side effects.     

暗黑链霉菌PY-1活性产物分析及其对葡萄霜霉病田间防效评价

为探究暗黑链霉菌Streptomyces atratus PY-1对葡萄霜霉病菌Plasomopara viticola的抑制作用,通过扫描电镜观察PY-1发酵滤液对葡萄霜霉病菌孢子囊的影响,检测PY-1的活性物质种类,并测定其对葡萄霜霉病的田间防效。结果表明,PY-1发酵滤液能够导致葡萄霜霉病菌孢子囊和孢子囊梗出现褶皱、破裂和畸形,进而使其丧失侵染功能。PY-1菌株代谢产物中包含几丁质酶、蛋白酶、嗜铁素、1-氨基环丙烷-1-羧酸脱氨酶、氰化氢、吲哚乙酸,不含纤维素酶。PY-1菌株对葡萄霜霉病具有较好的田间防效,发酵原液对葡萄霜霉病的田间中期防效可达到89.17%以上,末期防效达86.28%以上,比52.5%噁唑菌酮·霜脲氰2 000倍液的防效略低,但显著高于58%甲霜·锰锌1 000倍液的防效;PY-1菌株发酵液稀释700倍后对葡萄霜霉病的末期防效与甲霜锰锌1 000倍液防效相当。表明PY-1菌株具有研制防治葡萄霜霉病生防制剂的潜力。