Evaluation of breeding strategies for resistance and tolerance to late blight in potato by means of simulation

A field experiment with three potato cultivars, where plants were inoculated withPhytophthora infestans, was used to parameterize a model of potato growth and blight population dynamics. The model was validated by accurately simulating a field experiment conducted in another year. Sensitivity analysis with the model showed that late cultivars are longer able to maintain a green canopy in the presence of disease, but still suffer more yield loss than early cultivars. The level of partial resistance of a cultivar was more important than its level of tolerance, and other plant characteristics. The model calculations showed that only between 4 and 15% of the yield loss in the experiments was due to accelerated leaf senescence caused by the disease; the major part of the loss was caused by lesion coverage of leaves.     

Toxicity of propamocarb to the late blight fungus on potato

Propamocarb (Previcur-N; propyl-[3-dimethylamino-propyl] carbamate-monohydrochloride) was testedin vivo against 32 field isolates ofPhytophthora infestans from six countries. Fungicide dosages required to achieve 90% control of the blight ranged between 676 and 1530 ppm a.i. in potted potato (cv. ‘Alpha’) plants and between 1135 and 2648 ppm in potato tuber slices. Isolates from Israel were less sensitive to the fungicide than isolates from Europe or North America. Toxicity of propamocarb was not related to resistance or sensitivity to phenylamide fungicides (e.g. metalaxyl). Nevertheless, most metalaxyl-resistant isolates from Israel were less sensitive to propamocarb than most metalaxyl-sensitive isolates from this country. Monocyclic epidemics conducted with the 20 Israeli isolates in the field showed that 1081–2012 ppm of the fungicide was required to achieve 90% control of the disease. Laboratory experiments revealed that the fungicide was poorly active against sporangial germination and had a limited curative efficacy. It exhibited a translaminar translocation in leaves but a poor acropetal or basipetal systemicity from foliage. Propamocarb + mancozeb mixtures (1:1, v/w) were synergistically effective in controlling the blight. Growers in Israel use tank mixtures of propamocarb (Dynone) and mancozeb to combat late blight in potato fields where phenylamide-resistant isolates ofP. infestans are prevalent.     

十一种杀菌剂防治马铃薯晚疫病的适宜施用时期

为了明确当前市场上具有代表性的11种防治马铃薯晚疫病杀菌剂的适宜施用时期,采用人工接种马铃薯叶片的方法,测定了其中5种药剂在接种晚疫病菌前10 d内不同时间施用的预防效果和其中7种药剂在接种后24 h内不同时间施用的治疗效果。结果显示:在保证良好防治效果的前提下,药剂在病原菌接种前保护性施用的适宜时期比在接种后治疗性施用的适宜时期要长。在测试浓度下,接种前5种供试药剂保持100%防治效果的适宜施用时期为接种前3~10 d;而接种后7种供试药剂保持相同防效的适宜施用时期仅为接种后6~12 h,在生产实际中难以操作。研究结果说明,在田间马铃薯晚疫病的化学防治中,为了取得良好防治效果,每次用药均应在病菌侵入之前采用保护性施用,尽量避免在病菌侵入之后进行治疗性施用。     

四种化合物诱导马铃薯抗晚疫病的效果及其相关防御基因表达分析

为探究水杨酸、壳聚糖、麦角甾醇和纤维二糖4种化合物诱导马铃薯抗晚疫病的效果及其作用机理,采用整株喷雾法、挑战接种法、菌丝生长速率法分析4种化合物对马铃薯晚疫病的预防作用及对其病原菌致病疫霉Phytophthora infestans的离体抑制效果,利用实时荧光定量PCR技术测定4种化合物诱导后马铃薯病程相关蛋白和防御酶基因的调控表达情况。结果显示,水杨酸、壳聚糖、纤维二糖和麦角甾醇分别处理马铃薯植株24 h后接种致病疫霉,在多个浓度下对晚疫病均有不同程度的诱导抗性,其中在1 000 μmol/L浓度（壳聚糖浓度为1 000 mg/L）时诱导抗性最好,较对照显著提高,诱抗效果分别为54.73%、61.99%、59.73%和48.59%。壳聚糖对马铃薯晚疫病的诱抗效果高于其它3种化合物,且只有壳聚糖能显著抑制致病疫霉的菌丝生长和孢子囊形成,说明壳聚糖除了能诱导作物产生抗病性外,还具有直接抑菌作用。4种化合物诱导后马铃薯病程相关蛋白基因PR1、过氧化物酶基因POD和多酚氧化酶基因PPO的表达量在施用早期均显著升高。表明这些化合物能诱导马铃薯对晚疫病产生抗性,可能与马铃薯体内过氧化物酶、多酚氧化酶的活性和抗病信号传导途径关键基因PR1的表达有关。     

马铃薯晚疫病菌对甲霜灵抗性的快速检测方法

为快速检测马铃薯晚疫病菌——致病疫霉Phytophthora infestans对甲霜灵的抗性,基于已知致病疫霉RPA190基因的T1145A变异引起的F382Y氨基酸点突变与甲霜灵抗性有关,通过设计4对特异性引物F382Y-F1/F382Y-R、F382Y-F2/F382Y-R、F382Y-F3/F382Y-R和F382Y-F4/F382Y-R建立等位基因特异性PCR(allele specific-polymerase chain reaction,AS-PCR)快速分子检测法,其中F382Y-R引物在4对引物里保持不变,并分析所建分子检测法的特异性和灵敏度。结果表明,正向特异性引物F382Y-F2、F382Y-F3和F382Y-F4的3''末端在致病疫霉抗甲霜灵菌株原有核苷酸突变位点1145A(对应引物为F382Y-F1)的基础上,再人工引入第1144位的核苷酸突变,将1144T分别突变成1144G、1144C和1144A,并优化退火温度和特异性引物与内参引物ITS1/ITS4比例,最终确定最适退火温度分别为54、60和58℃,特异性引物与内参引物最适浓度比例均为5∶1。以该3条引物对应的3对特异性引物和内参引物ITS1/ITS4组成的3组多重AS-PCR引物对甲霜灵敏感和抗性菌株具有良好的特异性,敏感菌株的扩增产物含1条879bp的内参片段,抗性菌株的扩增产物为1条879bp的内参片段和1条461bp的目的片段。3组AS-PCR检测体系均具有较高的灵敏度,其中引物F382Y-F4/F382Y-R对致病疫霉DNA的检测灵敏度达0.4pg/μL,引物F382Y-F2/F382Y-R和F382YF3/F382Y-R的检测灵敏度达4pg/μL。     

Control of late blight in organic potato production: evaluation of copper-free preparations under field,growth chamber and laboratory conditions

In order to replace copper fungicides in organic potato production, 53 copper-free preparations (CFPs) based on natural compounds, including plant extracts and microorganisms, and five copper preparations were evaluated for their potential to control Phytophthora infestans, the pathogen that causes late blight of potatoes. In in vitro assays, 30% of the CFPs inhibited indirect germination of sporangia, 26% mycelial growth and in growth chamber experiments, 21% efficiently reduced foliar blight of tomato plants. In micro-plot field trials with applications twice a week, the copper preparations were the most effective and reduced foliar blight by 99%. Of the CFPs tested, Oekofluid P, Mycosin and other sulphuric clays, and C-2000 reduced late blight the most, from 63% to 37%. In small-plot trials in 2001, 2002 and 2004, 27 CFPs with different formulations and four copper preparations were examined. In 2004, copper preparations at full and reduced rates and sulphuric clays were applied either weekly or according to the decision support system Bio-PhytoPRE. With Bio-PhytoPRE, copper preparations reduced foliar blight of potatoes by 23–77% and increased tuber yield by 2–28%, depending on the copper rate applied and year. With CFPs, maximal efficacy was 17% and no effect on tuber yield was observed. In vitro and in vivo trials showed that the rainfastness and the persistence of CFPs was low compared with copper preparations. This indicates that the failure of CFPs under field conditions is probably due to a lack of stability under prevailing environmental conditions and not to a lack of efficacy. Until stable formulations for CFPs are developed, an optimised and restricted use of copper fungicides using a decision support system could help to control late blight in organic potato production and to reduce copper input into the environment.     

Photosynthesis is not impaired in healthy tissue of blighted potato plants

The net photosynthetic rates of green leaf tissue of potato plants of different cultivars were measured in the field and in a controlled environment after infection of the plants byPhytophthora infestans.Infection had no significant effect on the net photosynthetic rate at light saturation, the efficiency of light use at low light intensities, or dark respiration. The reported effect ofP. infestans on tuber yield seems to be caused solely by a reduction in the green leaf area. Therefore, a high rate of photosynthesis in green leaf tissue of infected plants is not a good selection criterion for potato genotypes.Samenvatting Bepalingen van netto fotosynthesesnelheden, bij verschillende aardappelrassen, werden uitgevoerd in het veld en onder geconditioneerde omstandigheden. De metingen werden gedaan aan groen blad van planten die in verschillende mate waren aangetast doorPhytophthora infestans.Infectie had geen significante invloed op de netto fotosynthesesnelheid bij lichtverzadiging, de efficiëntie van lichtbenutting bij lage lichtintensiteit, of de donkerademhaling. Het effect vanP. infestans op de knolopbrengst van aardappelrassen lijkt uitsluitend veroorzaakt te zijn door een vermindering van groen bladoppervlak. Daarom is selectie van aardappelgenotypen met superieure handhaving van fotosynthetische activiteit bij aantasting, geen kansrijk veredelingsdoel.     

Mycorrhiza induced resistance in potato plantlets challenged by Phytophthora infestans

Biological control of soil-borne pathogens by arbuscular mycorrhizal (AM) fungi has been repeatedly demonstrated. However, their role in the control of above-ground hemibiotrophic pathogens is less conclusive. Here, we investigated in vitro the impact of an AM fungus on Phytophthora infestans in potato plants. The leaf infection index was decreased in mycorrhizal potato plants. Real-Time Quantitative PCR revealed the induction of two pathogenesis related genes (PR1 and PR2) in the leaves of mycorrhizal plants shortly after infection with P. infestans. These results suggested a systemic resistance in mycorrhizal plants, related to the priming of the two PR genes in potato.     

Synergistic interaction between BABA and mancozeb in controlling<Emphasis Type="Italic">Phytophthora infestans</Emphasis> in potato and tomato and<Emphasis Type="Italic">Pseudoperonospora cubensis</Emphasis> in cucumber

Spray mixtures consisting of the plant activator BABA (DL-3-aminobutyric acid) and the protectant fungicide mancozeb were significantly more effective than BABA or mancozeb alone in controlling late blight (Phytophthora infestans) in potato and tomato and downy mildew (Pseudoperonospora cubensis) in cucumber. A mixture composed of 5 parts BABA and 1 part mancozeb (w/w, a.i.) exhibited a higher synergy factor than the 1+1 or the 1+5 (BABA + mancozeb) mixtures. No synergistic interaction was detected between BABA plus mancozeb in controlling sporangial or cystospore germination, nor mycelial growth ofP. infestans in vitro. The results showed enhanced effect of mancozeb in BABA-induced plants, suggesting, therefore, that lower dosages of this fungicide may be sufficient to control late blight or downy mildew under field conditions. http://www.phytoparasitica.org posting July 15, 2003.     

马铃薯晚疫病菌对氟吡菌胺抗性监测及9种常规药剂对马铃薯晚疫病田间防效评估

为探明北方一季作区马铃薯晚疫病菌(Phytophthora infestans)对氟吡菌胺抗性发生动态,2012～2016年自河北、内蒙古、辽宁、吉林、黑龙江等地区采集分离到马铃薯晚疫病菌520个菌株,采用菌丝生长速率法检测供试菌株对氟吡菌胺的敏感性,并以茎叶喷雾法评价9种生产上常用药剂对马铃薯晚疫病的田间防治效果.结...     

A High Multi-drug Resistance to Chemically Unrelated Oomycete Fungicides in Phytophthora infestans

Mutants of Phytophthora infestans with high resistance to the amidocarbamates iprovalicarb and benthiavalicarb and to the cyanoimidazole cyazofamid were isolated after UV-mutagenesis and selection on media containing one of the above fungicides. In vitro fungitoxicity tests showed that all resistant strains presented a highly reduced sensitivity to both cyazofamid and to the amidocarbamates. Cross-resistance studies with other oomycete fungicides from different chemical groups showed that the mutation(s) for resistance to iprovalicarb (IPV), benthiavalicarb (BVC) and cyazofamid (CZF) also greatly reduced the sensitivity of mutant strains to the phenylamide metalaxyl, acetamide cymoxanil, morpholine dimethomorph, benzamide zoxamide and to chlorothalonil. A lower reduction of sensitivity of mutant strains to the strobilurins azoxystrobin, kresoxim-methyl, pyraclostrobin and trifloxystrobin, azolones famoxadone and fenamidone and to antimycin A was observed. A resistance correlation was not apparent for the dithiocarbamate propineb and phenylpyridinamine fluazinam. Studies of fitness parameters in the wild-type and mutant strains of P. infestans showed that most resistant isolates had significantly reduced sporulation and sporangial germination, but not in the differentiation of sporangia into zoospores. Pathogenicity tests on tomato seedlings showed that most resistant isolates were significantly less pathogenic compared to the wild-type parent strain. However, experiments on the stability of the resistant phenotypes did not show a reduction in resistance when the mutants were grown for more than eleven generations on inhibitor-free medium. This is believed to be the first report of high level multi-drug resistance in fungal pathogens to chemically unrelated fungicides inhibiting different sites of cellular pathway.     

Restriction of potato and tomato late blight development by sub‐phytotoxic concentrations of boron

Boron is a microelement required for normal growth and development of plants but its positive effect is restricted to a narrow range of concentrations. The gradual increase in use of recycled water, which contains high concentrations of boron for irrigation, has already raised the level of boron in soils and plants in southern Israel. This research was conducted to examine the direct effects of sub‐phytotoxic boron concentrations on potato late blight epidemics and to explore the mode of action of boron against Phytophthora infestans. When boron was applied alone to field grown potato plants it did not affect the epidemic. However, together with a reduced rate of the fungicide Melody Duo (propineb + iprovalicarb), boron improved late blight suppression compared to plants treated with the fungicide alone. The ED₅₀ of boron against P. infestans (256·4 mg L^?1) was about 6400 times higher than the ED₅₀ value of the fungicide chlorothalonil (0·04 mg L^?1), indicating that boron does not have a direct fungicidal activity that would explain the level of protection seen in the field. In greenhouse experiments conducted with potted tomato plants, boron decreased late blight severity in both treated leaves and distant leaves not treated with boron. The results suggest that boron is active locally but also may induce systemic acquired resistance against P. infestans.     

棉铃疫病生防细菌筛选、鉴定及制剂防治效果

为有效防治由苎麻疫霉Phytophthora boehmeriae引起的棉铃疫病,采用平板对峙培养法、离体试验和田间小区试验对棉花、玉米、番茄、黄瓜等作物根围细菌进行筛选,结合形态学观察、16S rDNA序列分析及Biolog微生物自动鉴定系统对筛选出的生防细菌进行分类鉴定,并通过离体试验对优秀生防菌2种制剂的防治效果进行评价。拮抗细菌初筛试验结果表明,在供试的1 250株作物根围细菌中,399株细菌菌株有良好的抑菌能力,其中40株拮抗细菌菌株抑菌率在80.00%以上,菌株HMB22922抑菌能力突出,对棉铃疫病菌的抑菌率为85.71%,抑菌带宽10.0 mm。室内筛选试验结果表明,菌株HMB22922、HMB23917和HMB21405的病情指数显著低于空白对照,与化学药剂对照间差异不显著,人工接种苎麻疫霉5 d和7 d后,3株菌株对棉铃疫病的防治效果分别为73.92%、69.58%、56.54%和81.34%、66.67%、64.60%。田间筛选试验结果表明,菌株HMB22922培养液对棉铃疫病的田间防治效果为47.12%～58.64%,优于其它2株供试菌株,与化学药剂的防治效果相当且防治效果稳定。生防菌株HMB22922经形态学观察、序列分析和和微生物鉴定系统被鉴定为萎缩芽胞杆菌Bacillus atrophaeus。人工接种苎麻疫霉5、7和12 d时,以菌株HMB22922为有效成分的2.5′109 CFU/mL萎缩芽胞杆菌悬浮剂的防治效果分别为86.33%、78.66%和64.44%,均优于其相同菌量培养液的防治效果。     

Phosphite compounds reduce disease severity in potato seed tubers and foliage

Phosphites (Phi) are alkali metal salts of phosphorous acid, with the ability to protect plants against different pathogens. In this research, the effect of Phi applied to potato plants on severity of three important potato diseases in Argentina was assessed. Seed tubers and foliage of potato cvs Shepody and Kennebec were treated with Phi to assess effects on resistance against Phytophthora infestans, Fusarium solani and Rhizoctonia solani. Protection resulting from Phi treatment in seed tubers was high against P. infestans, intermediate against F. solani, and low against R. solani. In addition, seed tubers treated with calcium or potassium phosphites (CaPhi and KPhi, respectively) at 1% of commercial product emerged earlier than untreated ones. When Phi were foliarly applied two or four times at different doses, high levels of protection against P. infestans were achieved in both cultivars. Higher protection was observed in Kennebec when CaPhi was applied, while in Shepody this was true for KPhi. Expression of β-1,3-glucanases was induced at different times after treatment but no correlation between β-1,3-glucanases expression and foliar protection level was found. On the other hand, Phi positive protection effects did not produce negative effects in plant growth. Leaves from CaPhi-treated plants showed a darker green colour than leaves from control plants; also an increase in Rubisco protein and a delay in crop senescence was observed.     

Evaluation and analysis of reduction of late blight disease in a diploid potato progeny

The rate of late blight disease was analysed for individuals of a diploid Solanum phureja – Solanum tuberosum dihaploid hybrid population (PD), using three different assessment techniques, in the laboratory, screenhouse, and field. These hybrids expressed low disease rates in the field, comparable to resistance based on intact R genes. However, none of the parents of PD expressed any R genes and the pattern of segregation within the PD population was not indicative of R-gene inheritance. The foliage (or leaflet) area diseased had the largest broad-sense heritability of all criteria analysed, in all tests performed. In the field evaluations, the PD population showed intermediate levels of broad-sense heritability for foliage area diseased, relative to the much larger heritability detected for the group of controls possessing R genes. Resistance in the field of the PD hybrids had very little genotype-environment (G × E) interaction, indicating stability of its expression. All genotypes without R genes exhibited heritable, reduced rates of late blight disease in the field, but they were susceptible with low heritability in screenhouse and laboratory tests. This differential expression of disease indicates that the plants' indirect response to unknown environmental stress in the field may have been measured. The value of the PD hybrids for breeding of late blight resistant potato and the use of the disease rate data for detection of the underlying quantitative trait loci are discussed.     

Integrated control of potato late blight: predicting the combined efficacy of host resistance and fungicides

Integrating cultivars that are partially resistant with reduced fungicide doses offers growers an opportunity to decrease fungicide input but still maintain disease control. To use integrated control strategies in practice requires a method to determine the combined effectiveness of particular cultivar and fungicide dose combinations. Simple models, such as additive dose models (ADM) and multiplicative survival models (MSM), have been used previously to determine the joint action of two or more pesticides. This study tests whether a model based on multiplicative survival principles can predict the joint action of fungicide doses combined with cultivars of differing partial host resistance. Data from eight field experiments on potato late blight (Phytophthora infestans) were used to test the model; the severity of foliar blight was assessed and scores used to calculate the area under the disease progress curve (AUDPC). A subset of data, derived from the most susceptible cultivar, King Edward, was used to produce dose–response curves from which parameter values were estimated, quantifying fungicide efficacy. These values, along with the untreated values for the more resistant cultivars, Cara and Sarpo Mira, were used to predict the combined efficacy of the remaining cultivar by fungicide dose combinations. Predicted efficacy was compared against observations from an independent subset of treatments from the field experiments. The analysis demonstrated that multiplicative survival principles can be applied to describe the joint efficacy of host resistance and fungicide dose combinations.     

寡雄腐霉GAQ1对辣椒疫病的防效及对辣椒的促生作用

为明确自主分离的生防菌株寡雄腐霉Pythium oligandrum GAQ1对辣椒疫病的生防效果及其防御机制,通过平板拮抗和盆栽防效试验测定寡雄腐霉菌株GAQ1对辣椒疫霉Phytophthora capsici菌丝的拮抗作用、对辣椒疫病的防效和对辣椒的促生效果,同时应用实时荧光定量PCR技术检测菌株GAQ1处理后辣椒抗性基因表达的变化。结果表明,寡雄腐霉菌株GAQ1的菌丝可以缠绕并吸附寄生在辣椒疫霉菌丝表面或穿入菌丝体内,使辣椒疫霉菌丝细胞死亡;菌株GAQ1发酵液处理辣椒离体叶片再接种辣椒疫霉后产生的病斑直径较对照组显著减少,离体防效为30.79%;接种菌株GAQ1菌丝球后,辣椒疫病的病情指数较对照组显著降低,盆栽防效达69.16%;经菌株GAQ1处理辣椒后可诱导相关抗性基因PR1、WRKY40、WRKY53、ACCO和GST的相对表达量出现不同程度的升高,说明菌株GAQ1可诱导辣椒植株产生不同程度的防御系统应答。菌株GAQ1对辣椒具有良好的促生效果,处理后第5周其株高、株重及根重分别较对照组提高10.11%、33.23%和24.72%,其叶片中叶绿素a、叶绿素b及类胡萝卜素的含量分...