Effect of Sexual Recombination on Pathotype Frequencies in Barley Powdery Mildew Populations of Artificially Inoculated Field Plots

Changes in pathotype frequencies due to sexual reproduction during summer were assessed in barley plots inoculated at the start of the growing season with powdery mildew isolates of the pathotype GL1 (Va22Vh) which belonged to a clonal lineage frequent in Northern France in 1992–1996 and was absent or rare in the local population at the time of the study. The capacity for recombination among isolates belonging to the pathotype GL1 was confirmed by performing crosses in the greenhouse. The field experiment was repeated over two years, with two field plots inoculated with either a single isolate (one mating type) or four isolates (two mating types). Following artificial inoculation, the frequency of GL1 increased to between 40% and 80% of the total conidia population at the end of the asexual epidemics before summer. When only one isolate (one mating type) of the pathotype GL1 was present, the frequency of the GL1 pathotype decreased to 4–5% of the ascospore population, following sexual recombination between the inoculated isolate and the local natural population. When isolates of both mating types of GL1 were inoculated, however, the GL1 pathotype remained dominant (28%) in the ascospore population. A pathotype dominant at the end of the summer might possibly have over-summered through asexual reproduction, or alternatively it may originate from sexual reproduction. The observed GL1 frequencies and numbers of virulences per isolate (complexity) observed on the field plots were compared to those calculated with a model assuming random or non-random mating.     

Cytological events in tomato leaves inoculated with conidia of <Emphasis Type="Italic">Blumeria graminis</Emphasis> f. sp. <Emphasis Type="Italic">hordei</Emphasis> and <Emphasis Type="Italic">Oidium neolycopersici</Emphasis> KTP-01

Leaves of tomato and barley were inoculated with conidia of Blumeria graminis f. sp. hordei race 1 (R1) or Oidium neolycopersici (KTP-01) to observe cytological responses in search of resistance to powdery mildew. Both conidia formed appressoria at similar rates on tomato or barley leaves, indicating that no resistance was expressed during the prepenetration stage of these fungi. On R1-inoculated tomato leaves, appressoria penetrated the papillae, but subsequent haustorium formation was inhibited by hypersensitive necrosis in the invaded epidermal cells. On the other hand, KTP-01 (pathogenic to tomato leaves) successfully developed functional haustoria in epidermal cells to elongate secondary hyphae, although the hyphal elongation from some conidia was later suppressed by delayed hypersensitive necrosis in some haustorium-harboring epidermal cells. Thus, the present study indicated that the resistance of tomato to powdery mildew fungi was associated with a hypersensitive response in invaded epidermal cells but not the prevention of fungal penetration through host papilla.     

陕西省小麦白粉菌群体毒性结构及遗传多样性

为明确近年来陕西省小麦白粉菌Blumeria graminis f. sp. tritici群体毒性及遗传变异情况,利用32份已知抗白粉病基因小麦品种(系)对2013年和2014年陕西省小麦白粉菌群体毒性结构进行测定,并应用扩增片段长度多态性(amplified fragment length polymorphism,AFLP)标记对2014年陕西省西安市、咸阳市、渭南市、宝鸡市及陕西省毗邻的甘肃省天水市5个小麦白粉菌地理群体共118株白粉菌菌株进行遗传多样性分析。毒性监测结果显示,供试小麦白粉菌群体对Pm1c、Pm2、Pm3d、Pm4a、Pm4b、Pm5b、Pm13、Pm21、Pm24、Pm30、Pm2+6、Pm2+Mld、Pm2+6+?、Pm4b+5b、Pm4+?、Pm5+6的平均毒性频率在0～17.23%之间,表明这些基因抗性保持较好;对Pm1a、Pm3b、Pm3c、Pm3e、Pm3f、Pm7、Pm8、Pm19、Pm1+2+9的平均毒性频率介于69.17%～99.60%之间,表明这些基因抗性已丧失。用筛选出的6对AFLP标记共扩增出831个多态性位点,多态性位点百分比为94.86%;小麦白粉菌群体遗传多样性指数和香农信息指数分别为0.1151和0.2036,遗传变异主要来源于群体内变异。群体间基因流为4.7939,表明5个小麦白粉菌群体间基因交流广泛。群体间遗传距离和地理距离两者显著正相关。     

不同机制杀菌剂对小麦白粉病的敏感性及与三唑酮的交互抗性

为明确三唑酮和氟环唑、吡唑醚菌酯、啶酰菌胺、嘧菌环胺、乙嘧酚5种不同作用机制的杀菌剂对小麦白粉病的敏感性及交互抗性,采用田间小区试验和室内喷雾离体叶段法测定了不同杀菌剂对小麦白粉病的防治效果。结果表明,5种不同作用机制杀菌剂对小麦白粉病的防治效果可达90%以上,而三唑酮最高的防治效果仅为72.17%;小麦白粉病菌群体对氟环唑、吡唑醚菌酯、啶酰菌胺、嘧菌环胺、乙嘧酚的敏感性EC50分别在0.087~1.901、0.058~1.402、0.186~3.014、0.222~6.005、0.006~1.742μg/mL之间,5种不同作用机制杀菌剂的敏感性均呈连续单峰曲线,可作为小麦白粉病菌对5种不同作用机制杀菌剂的敏感基线。研究表明,三唑酮与氟环唑、吡唑醚菌酯、啶酰菌胺、嘧菌环胺、乙嘧酚之间不存在交互抗性。     

大麦白粉菌种群毒性监测及抗性材料鉴定

2005和2006年从我国冬大麦区采集和分离大麦白粉菌单胞菌株729个,利用Pallas近等基因系进行致病型鉴定和群体毒性频率分析.同时,利用分离得到的不同致病型菌株,通过抗谱分析的方法鉴定了328份大麦品种(系)的白粉病抗性和抗病基因.结果显示:大麦白粉菌群体对抗病基因Mlal Mla(A12)、Mla3、Mla6 Mla14、Mla7 Mla(No3)、Mla7 Ml(Lg2)、Mla9 Mlk、Mla9、Mlal3 MlaRu3、Mlpl、Mlg(Cp)和mlo5的毒性频率为0;对Mla12 MlaEm2、Mla7 Mlk、Mlat Mla8、Mla10MlaDu2和Mlk1的毒性频率很低,分别为0.1%、0.4%、0.9%、2.8%和4.2%.两年共鉴定出不同的致病型21个,致病型000、001和003在两个年度皆为优势致病型.所鉴定的328份材料绝大多数感病,仅37份抗病材料,能明确推导出抗白粉病基因的品种(品系)很少,这些品种(品系)含有的抗白粉病基因为Mr(Bw)Mla8、Mlg、Mira Mla8、Mla9 Mla1、Mla Mla(A12)和mlo5.     

Antioxidant,ethylene and membrane leakage responses to powdery mildew infection of near-isogenic barley lines with various types of resistance

Leaves of powdery mildew-susceptible barley (Hordeum vulgare cv. Ingrid) and related near-isogenic lines bearing various resistance genes (Mla12, Mlg or mlo5) were inoculated with Blumeria graminis f. sp. hordei race A6. Fungal attack induced several-fold increases in ethylene emission and electrolyte leakage in leaves of susceptible Ingrid beginning 3 days after inoculation. Activities of peroxidase, superoxide dismutase, glutathione S-transferase, ascorbate peroxidase and glutathione reductase enzymes were induced markedly in susceptible leaves 5–7 days after inoculation. Similar, but less pronounced pathogen-induced changes were detected in inoculated leaves of Mla-type resistant plants that show hypersensitive cell death upon inoculation, and, to an even lesser extent, in the Mlg and mlo lines, where no visible symptoms accompanied the incompatible interaction. Glutathione content increased only in susceptible barley 7 days after inoculation. Catalase activity, total ascorbate content and redox state were not influenced by inoculation in any of the genotypes. The activity of dehydroascorbate reductase was significantly reduced 3–5 days after inoculation in the susceptible parental plants and after 5 days in Mla and Mlg lines, while it was stable in the mlo barley. Slightly elevated levels of H₂O₂ were observed in the inoculated resistant plants. In contrast, H₂O₂ content decreased in the susceptible line 7 days after pathogen attack. These data indicate that high levels of antioxidants are involved in the compatible interaction of susceptible barley and powdery mildew by protecting the pathogen from oxidative damage.     

小麦白粉病菌对喹氧灵和苯菌酮的敏感基线及药剂的生物活性

采用离体叶段法,分别测定了从河北、河南、湖北、陕西和四川5省分离的53个小麦白粉病菌单孢菌株对苯菌酮和喹氧灵的敏感性,并分析了白粉病菌对三唑酮和苯菌酮以及喹氧灵之间的 交互抗性。结果表明:小麦白粉病菌群体对苯菌酮和喹氧灵的平均EC50值分别为(0.001 9±0.000 6) 和(0.013 1±0.002 0) mg/L,苯菌酮比喹氧灵具有更高的抑菌活性;小麦白粉病菌对三唑酮和苯菌酮与喹氧灵之间均不存在交互抗性(R2值分别为0.102 6和0.491 9);室内盆栽试验结果显示:接种前1 d和接种后1d施药,苯菌酮和喹氧灵对小麦白粉病的保护与治疗作用防效分别为92.21%、84.25%和82.43%、70.25%,表明这2种药剂不仅具有优异的保护作用,同时还具有较好的治疗作用。     

Factors affecting build-up of fungicide resistance in powdery mildew in spring barley

In replicate-plot field experiments done in the UK, at one site in Avon for 3 years and another in Warwickshire for 2 years, application of ethirimol or triadimenol sprays or seed treatments was followed by decreases in sensitivity of mildew samples to the particular fungicide applied. Application of ethirimol-triadimenol or tridemorph-triadimenol mixtures caused smaller or no decreases in sensitivity. Differences between isolates in responses to ethirimol and to triadimenol treatment were usually negatively correlated. Early-season inoculum differed in fungicide sensitivity between sites. At one site sensitivity shifted markedly from one season to another. No clear interactions between cultivar, mildew pathotype and shifts in fungicide response could be discerned. There were no major differences in resistance build-up between seed or spray treatments.In veldexperimenten met herhalingen uitgevoerd in Engeland, gedurende een periode van 3 jaar in Avon en gedurende een periode van 2 jaar in Warwickshire, werd na behandeling van zomergerst met ethirimol of triadimenol een verminderde gevoeligheid van meeldauw voor deze fungiciden waargenomen.Behandelingen met mengsels van ethirimol-triadimenol of tridemorf-triadimenol gaven weinig tot geen verminderde gevoeligheid. Verminderde gevoeligheid van isolaten voor ethirimol was meestal gecorreleerd met een verhoogde gevoeligheid voor triadimenol en omgekeerd. Vroeg in het seizoen werd in de meeldauwpopulatie op de twee proefvelden een verschil in gevoeligheid voor de fungiciden waargenomen. Op één proefveld trad van het ene op het andere seizoen een aanzienlijke verandering in de gevoeligheid voor de fungiciden op. Er was geen duidelijke correlatie tussen de waargenomen verminderde gevoeligheid voor de fungiciden en de gebruikte cultivars of voorkomende fysio's. Verminderde gevoeligheid voor de fungiciden werd zowel bij zaadbehandeling als bij het bespuiten van planten waargenomen.     

A comparative study of resistance to powdery mildew in wild emmer wheat in the seedling and adult plant stage

An isometric virus was isolated from cucumber plants growing in a plastic house in Crete and showing stunting and bright yellow mosaic of the leaves. Based on host range, properties in crude sap, behaviour during purification, electron microscopy and serology, the virus was identified as an isolate of artichoke yellow ringspot nepovirus. Ecological data corroborate transmission of the virus via the soil.Samenvatting Uit komkommerplanten in plastic-foliekassen op Kreta werd een bolvormig virus geïsoleerd; de aangetaste komkommerplanten vertoonden dwerggroei en helder geel mozaïek op de bladeren. Gebaseerd op de resultaten verkregen uit onderzoek met het virus naar de waardplantenreeks, de eigenschappen in perssap, zuivering, elektronen-microscopie en serologie kon het virus worden geïdentificeerd als een strain van het artichoke yellow ringspot nepovirus. Waarnemingen op het gebied van de ecologie wijzen op overdracht van het virus via de grond.     

抗苯醚菌酯(ZJ0712)小麦白粉病菌突变体的诱导及其生物学特性

经过药剂驯化10代后,从对苯醚菌酯(ZJ0712)敏感的小麦白粉病菌Blumeria graminis f.sp.tritici 3个菌株中获得3个抗药突变体,突变体的抗性指数均大于80,且其抗性能够通过无性繁殖稳定遗传。室内接种试验发现,突变体的致病力与亲本菌株无明显差异。细胞色素b (cyt b)基因片段序列分析发现,突变体cyt b基因的第143位密码子均由敏感菌株的GGT(丙氨酸)突变成了GCT(甘氨酸)。研究结果表明,小麦白粉病菌对苯醚菌酯存在较高的抗性风险,该药剂在使用过程中需注意采取相应的抗性治理措施,以延缓抗性发生。     

Virulence and diversity of <Emphasis Type="Italic">Blumeria graminis</Emphasis> f.sp. <Emphasis Type="Italic">hordei</Emphasis> in East China

Four hundred and sixty-one isolates of Blumeria graminis f.sp. hordei were obtained from eight populations occurring on cultivated barley (Hordeum vulgare) at four geographically distant locations in China during 2003 and 2004. Their virulence frequency was determined on 30 differential lines. No isolate was virulent on differential lines possessing the resistance genes Mla1, Mla3, Mla6, Mla7, Mla9, Mla12, Mla13, Mlat, Mlg, Mla10, Mla22, Mla23, Mlp1, Ml(N81) and Mlmw. Virulences to the first nine resistance genes are prevalent in Europe and constitute the main part of genetic distance between Chinese and European populations. Conversely, no isolate was avirulent on the differential lines possessing the genes Mla8 and Ml(Ch). The frequencies of isolates overcoming the genes Mla2, Mla11, Mlk1 and Mlk2 were .4–9.3%, and frequencies of isolates overcoming the genes Mlh, MlLa, Ml(Bw), Mlra, Ml(Ru2), mlw, MlGa, MlWo and Mlnn ranged from 18.2% to 98.7%. Based on reactions of differential lines possessing the genes Mlk1, Mlh, MlLa, Ml(Bw), Mlra and Ml(Ru2), pathotypes were identified and diversity parameters calculated. Eleven of 22 detected pathotypes were found in both years and comprised 94.6% of isolates. Generally, the populations from different locations in 1 year were more closely related than populations collected from the same locations in different years. Complete effectiveness of the resistance genes, for which no corresponding virulences were found, will allow Chinese breeders to access many modern European barley cultivars that are fully resistant to powdery mildew in China, including those possessing the non-host resistance gene mlo.     

小麦白粉菌ISSR分子标记体系构建及其分离菌株的多样性分析

为了对小麦白粉菌群体的多样性进行研究,构建了其ISSR分子标记体系。以小麦白粉菌基因组DNA为模板,用正交设计和单因素水平优化的方法对ISSR反应程序中的一些重要参数进行摸索和优化,建立了小麦白粉菌ISSR-PCR反应的优化反应体系;对20个ISSR引物的退火温度进行了优化,并筛选出一些多态性较好的ISSR引物。对33个分离菌株的ISSR扩增表明,ISSR标记在我国小麦白粉菌中存在较高的多态性;对ISSR标记揭示的白粉菌的遗传多样性和毒性多样性进行了比较,结果表明两者之间存在一定程度的相关性。     

2013年我国部分麦区小麦白粉菌群体对温度敏感性及其与毒性的关系

 研究小麦白粉病菌群体对温度的敏感性及其与毒性的关系,可为气候变化背景下小麦白粉病的长期发生趋势预测和防治提供依据。本研究对2013年采自云南、四川、河南、北京和陕西五省（市）的小麦白粉菌标样进行分离纯化,获得90个单孢子堆分离物,采用小麦离体叶段法进行温度敏感性测定,并利用32个已知抗病基因品种（系）进行毒性测定,且对两者之间的相关性进行分析。结果表明,供试菌株ET50分布于19.78℃~25.15℃,平均ET50为23.22℃,其中73.34%的供试菌株ET50值介于22℃到24℃之间,小于22℃的菌株占10%,大于24℃的菌株占16.67%。利用Popgen1.32软件对5个省（市）小麦白粉病菌群体的毒性进行多样性分析,结果表明五省市小麦白粉菌群体毒性水平上的多样性指数 H 值为0.237 0,其中四川群体毒性多样性水平最高,Nei’s基因多样性指数 H 值为0.221 2,河南群体最低, H 值为0.197 5。小麦白粉菌群体对温度敏感性与毒性多样性相关性分析结果表明,ET50与毒性多样性（ H 和 I 值）间存在显著的负相关。     

2010-2017年河南省小麦白粉菌群体的毒性变化动态监测

为了解河南省小麦白粉菌群体毒性结构及年度动态变化, 对河南省近8年来的小麦白粉菌群体毒性进行了监测, 结果表明其病菌群体对小麦白粉病抗性基因或组合Pm1a、Pm3a、Pm3b、Pm3c、Pm3f、Pm3e、Pm5a、Pm6、Pm7、Pm8、Pm17、Pm19、Pm1+2+9的毒性频率高于70%, 这些基因为无效抗病基因; 对抗性基因或组合Pm1c、Pm2、Pm5b、Pm12、Pm13、Pm16、Pm21、Pm22、Pm23、Pm24、Pm35、Pm36、Pm46、Pm“XBD”、Pm2+Mld、Pm2+6的毒性频率低于30%, 其中对抗性基因Pm12、Pm16和Pm21的毒性频率为0%, 这些基因依然为有效的抗病基因。病菌群体对Pm4a、Pm4b、Pm4+8等抗性基因或组合的毒性频率近年来总体呈上升趋势, 且毒性频率已达到30%~70%, 应加强对这些抗病基因或组合的重点监测, 并建议在抗病育种中谨慎使用。     

四川西南部小麦白粉菌群体毒性及遗传多样性分析

为明确四川西南部白粉菌群体毒性及其遗传变异情况,本文将2011年采自四川西南部的小麦白粉病标样进行单孢子堆分离纯化,共获得48个白粉菌菌株,并将其按采集地划分为4个地理群体。利用28份已知抗白粉病基因材料测定了群体的毒性频率,并运用SRAP分子标记技术探讨了其遗传多样性。毒性测定结果表明,白粉菌群体毒性较强,群体间毒性结构存在差异。供试群体对Pm1、Pm3a、Pm3b、Pm3d、Pm5a、Pm6、Pm19的平均毒性频率达50%以上,而Pm13、Pm XBD、Pm5b、Pm2+6、Pm5+6抗性保持良好,平均毒性频率在10%以下。群体间毒性遗传距离与地理距离之间有一定的相关性,但未达到显著水平。用筛选出的10对SRAP引物共获得160个清晰、稳定的扩增位点,多态性频率为50.63%;白粉菌群体遗传多样度(h)、Shannon信息指数(I)分别为0.198 8、0.322 7,遗传变异主要源于群体内部。群体间基因流数值均在6.50以上,说明四川西南部白粉菌群体间菌源迁移频繁,基因交流较为充分。M antel Test分析表明SRAP标记解释的群体遗传多样性与地理距离、毒性多样性间的相关性未达到显著水平。     

Variation in sensitivity to fungicides which inhibit ergosterol biosynthesis in wheat powdery mildew

Fungicides which inhibit ergosterol biosynthesis have been in use for control of wheat powdery mildew (Erysiphe graminis f. sp.tritici) in the Netherlands since 1978. Mildew populations were tested for their variation in sensitivity to triadimefon from 1982 to 1984.In 1982 isolates from the province Limburg, with a triazole spray-regime history, were less sensitive to triadimefon than isolates from the provinces Gelderland and Noord-Brabant, where triazoles had not been used. In the following years isolates with reduced sensitivity were also detected in the latter provinces and other parts of the country. This spread correlates with the increased use of triazoles, both in frequency and space, from 1983 onwards. The reduced sensitivity can as a whole or in part be responsible for the decline in field performance of triazoles, observed during these years.Cross-sensitivity to the triazoles triadimefon and propiconazole was established, but not to triazoles and the morpholine fungicide fenpropimorph. Effectiveness of the latter compound was similar to all isolates from Limburg tested in 1984. Field performance of fenpropimorph, introduced in 1983, appeared to be normal. It is recommended to counteract further development of resistance by sequential use of fenpropimorph early in the season (May) and triazoles at the end (June–July).Samenvatting Fungiciden die de ergosterolbiosynthese remmen worden in Nederland sinds 1978 gebruikt bij de bestrijding van tarwemeeldauw (Erysiphe graminis f. sp.tritici) Meeldauwpopulaties werden getoetst op hun variatie in gevoeligheid voor triadimefon van 1982 tot 1984.In 1982 bleken isolaten afkomstig uit de provincie Limburg, waar voordien triazolen werden toegepast, minder gevoelig te zijn voor triadimefon dan isolaten uit de provincies Gelderland en Noord-Brabant, waar nog geen triazolen werden gebruikt. In de daaropvolgende jaren werden isolaten met een verminderde gevoeligheid ook in laatstgenoemde provincies en in andere delen van het land gevonden. Deze uitbreiding is gecorreleerd met een toename in het gebruik van triazolen vanaf 1983 in areaal en frequentie. De afname in gevoeligheid kan geheel of gedeeltelijk verantwoordelijk zijn voor de verminderde meeldauwwerking van triazolen die gedurende deze jaren werd waargenomen.Kruisgevoeligheid werd vastgesteld voor de triazolen triadimefon en propiconazool, maar niet voor triazolen en het morfoline-derivaat fenpropimorf. De werking van dit middel was tegen alle isolaten uit Limburg die in 1984 werden getoetst, gelijk. De meeldauwwerking van fenpropimorf, dat in 1983 werd geïntroduceerd, was normaal. Aanbevolen wordt om verdere resistentie-ontwikkeling tegen te gaan door afwisselend gebruik van fenpropimorf vroeg in het groeiseizoen (mei) en een triazool aan het eind (juni-juli).     

2009年我国部分麦区小麦白粉病菌群体对温度的敏感性研究

 用小麦离体叶段法,设置22、23、24、25和26℃共5个温度处理,对2009年采自四川、北京、甘肃、河南、河北、陕西和湖北七省（市）的小麦白粉菌标样分离获得的129个单孢堆菌株对温度的敏感性进行了测定。结果表明,供试的129个菌株平均ET₅₀为22.49℃,其中ET₅₀最高为24.48℃,最低为19.88℃,供试菌株中40.52%的ET₅₀介于23℃和24℃之间,占参试菌株的近一半,7.63%的供试菌株ET₅₀≥24℃。不同省（市）的病菌群体对温度的敏感性存在一定差异,且病菌群体对温度的敏感性差异与经纬度存在显著或极显著的负相关关系。研究结果获得了温度对此病害的终止阈值平均为26.65℃,且对温度相对不敏感菌株在较高温度下的潜育期较短、病害的严重度较高。不同敏感性菌株的频率分布结果表明,自然环境中病菌群体已受到温度的选择压力。     

2004—2005年北方部分麦区白粉病菌小种动态及流行相关区品种抗性分析

对2004和2005年采自东北春麦区及山东、河北冬麦区的小麦白粉病110个标样进行生理小种鉴定和群体毒性频率测定;同时,测定了上述地区及与上述地区白粉病流行密切相关区的91份重要的生产和后备品种(系)的抗性及基因。结果表明:2004和2005年东北春麦区,2005年山东、河北春麦区小麦白粉病菌优势小种均为15号小种,出现频率为13.7%～16.0%,其次为415号和11号小种,出现频率为7.8%～12.0%。2004和2005年小麦白粉病毒力频率分析中,V3a、V3b、V3c、V5、V7、V8和V17的毒力频率均高于73.5%;V2、V4、V12、V16、V19和V21的毒力频率最低,为0～27.4%。上述白粉病流行相关区生产和后备品种(系)总体抗性不强,抗性较强的品种(系)推导出抗病基因主要为Pm21、Pm2 6及未知复合抗性基因等。