小麦新品系抗白粉病基因分析

 本文对6个小麦新品系所含的抗白粉病基因进行了遗传分析。将感病品种Liaochun10分别与SM 20121、SM 203390、SM 20125、SM 200332、SM 20126、SM 20005杂交和自交,并将这6个品系互配成半双列杂交组合。用小麦白粉菌15号小种的单孢堆菌系对各杂交组合的亲本、F₁、F₂代群体及F₃代家系进行了苗期抗病性鉴定。遗传分析表明,供试的6个品系对小麦白粉菌15号小种的抗性均由1对显性基因控制。等位性分析推断:SM 20121、SM 203390、SM 20125和SM 200332含有抗白粉病基因Pm12;SM 20126含有抗白粉病基因Pm21;SM 20005含有抗白粉病基因Pm16。建议将这6个品系作为优良抗病亲本利用。     

小麦慢白粉品种的抗性组分研究

 2003~2004年度和2004~2005年度在田间测定了16个小麦品种(系)对小麦白粉菌E20菌株的抗性组分,包括潜育期、侵染几率、产孢量、病斑扩展和传染期。结果表明,小麦慢白粉品种(系)与高感对照品种相比,表现为潜育期长、侵染几率低、产孢量低、病斑扩展慢的特点,且这4个抗性组分与对照的差异都达到了显著和极显著水平,但慢白粉品种的传染期变化无一定规律。参试慢白粉品种的抗性组分参数与病情曲线下面积(AUDPC)的相关分析表明,潜育期与AUDPC呈负相关,但相关性不显著;产孢量和病斑扩展与AUDPC呈正相关,两年数据相关性均达到显著水平;侵染几率与AUDPC呈正相关,但2003-2004年度相关性不显著,而2004~2005年度相关性显著;传染期与AUDPC相关关系不明显。由此可见,在慢白粉品种的病害流行中起主要作用的是产孢量、病斑扩展,而侵染几率、潜育期和传染期作用相对较小。     

Quantitative evolution of aggressiveness of powdery mildew under two-cultivar barley mixtures

The effects of cultivar mixtures on the evolution of aggressiveness of barley powdery mildew ( Blumeria graminis f.sp . hordei ) were modelled. It was found that the rate and direction of evolution of pathogen aggressiveness in a race-non-specific system and value at equilibrium, depends on initial resistance levels, proportions of component cultivars, autodeposition rates, the relative magnitude of the benefit of autoinfection, and the cost of alloinfection of spores. In the model, mixing cultivars in any proportions tended to reduce the aggressiveness of pathogens at equilibrium compared with pure stands, but this effect decreased when two mixture components were extremely unbalanced in proportion. Under low and medium autodeposition rates, the best control of the evolution of the pathogen was achieved by mixing two components in roughly equal proportions. The magnitude of aggressiveness at equilibrium increased as autodeposition rates increased. Though the level of initial resistance of mixture components did not have an impact on the aggressiveness of pathogens at equilibrium, it strongly influenced the transient values of aggressiveness and therefore the total amount of disease caused over an evolutionary period. The cost to the pathogen of alloinfection and benefit of autoinfection per se did not affect the final level of aggressiveness, but did affect the time to reach equilibrium. However, the ratio of the cost to the benefit did influence the final aggressiveness of the pathogen.     

50个小麦生产及后备品种(系)的抗白粉病基因推导

为明确我国小麦品种(系)中抗白粉病基因的组成,利用25个不同毒性的小麦白粉菌菌株对50个小麦生产及后备品种(系)进行抗白粉病基因推导,结果表明,参试的50个小麦品种(系)中有8个小麦品种(系)对供试的25个菌株全部感病,5个品种含有抗病基因Pm8,2个品种含有Pm4a,9个品种含有Pm2+6,4个品种含有Pm2,22个品种(系)可能含有供试基因之外的其他抗性基因或新基因。此研究结果可为小麦抗病育种以及品种利用提供依据。     

Local and systemic effects of oxylipins on powdery mildew infection in barley

Treatment of the first leaves of barley seedlings with the oxylipin colneleic acid, or the two trihydroxy oxylipins 9,12,13-trihydroxy-11(E)-octadecenoic acid and 9,12,13-trihydroxy-10(E)-octadecenoic acid, reduced infection of that leaf by the powdery mildew fungus Blumeria graminis Speer f sp hordei Marchal. When applied to first leaves, etheroleic acid and colneleic acid, as well as the trihydroxy oxylipin 9,12,13-trihydroxy-10(E),15(Z)-octadecadienoic acid, also reduced mildew infection in second leaves. In all cases where local and systemic effects against mildew were observed, activity of the defence-related enzyme phenylalanine ammonia lyase (PAL) was increased, but only following challenge inoculation with powdery mildew. Peroxidase activity was not affected by oxylipin treatment or mildew inoculation. Whether the effects observed were due to the oxylipins or to breakdown products is not known, since no information is available on the stability of these particular oxylipins on leaf surfaces. Nevertheless, these data represent the first report of systemic effects against pathogen infection following pre-treatment with oxylipins.     

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。     

不同杀菌剂对小麦白粉病菌室内毒力测定及混配增效药剂筛选

本研究旨在研究三唑类药剂戊唑醇、苯并咪唑类药剂多菌灵和甲氧基丙烯酸酯类吡唑醚菌酯对小麦白粉病菌的毒力及最佳配比筛选, 通过比较对2个不同品种小麦的防效差异, 为大田小麦白粉病的防治提供参考依据。采用盆栽法, 调查小麦叶片发病情况, 计算病情指数, 测定3种药剂对2个小麦品种上白粉病菌的毒力及其不同配比的共毒系数(CTC), 找出合理配比。结果表明在‘山农16’和‘泰农18’2个小麦品种上多菌灵与戊唑醇配比5∶3时, 增效作用均为最优, 共毒系数CTC分别为122.66和123.56; 戊唑醇与吡唑醚菌酯配比在2∶1时, 增效作用均为最优, CTC为139.09和129.97; 多菌灵与吡唑醚菌酯配比1∶1时, 增效作用均为最优, CTC为135.15和145.24。     

Mining wild barley for powdery mildew resistance

Powdery mildew, caused by Blumeria graminis f. sp. hordei (Bgh), is a worldwide disease problem on barley (Hordeum vulgare) with potentially severe impact on yield. Historically, resistance genes have been identified chiefly from cultivated lines and landraces; however, wild barley (H. vulgare subsp. spontaneum) accessions have proven to be extraordinarily rich sources of powdery mildew resistance. This study describes the characterization of a collection of 316 wild barley accessions, known as the Wild Barley Diversity Collection (WBDC), for resistance to powdery mildew and the genetic location of powdery mildew resistance loci. The WBDC was phenotyped for reaction to 40 different Bgh isolates at the seedling stage and genotyped with 10 508 molecular markers. Accessions resistant to all 40 isolates of Bgh were not found; however, three accessions (WBDC 053, 085 and 089) exhibited resistance to 38 of the isolates. Gene postulation analyses revealed that many accessions, while resistant, contained none of the 12 genes present in the Pallas near‐isogenic lines Mla1, Mla3, Mla6, Mla7, Mla9, Mla12, Mla13, Mlk1, MlLa, Mlg, Mlat and Ml(Ru2), suggesting that the accessions carry novel genes or gene combinations. A genome‐wide association study of powdery mildew resistance in the WBDC identified 21 significant marker‐trait associations that resolved into 15 quantitative trait loci. Seven of these loci have not been previously associated with powdery mildew resistance. Taken together, these results demonstrate that the WBDC is a rich source of powdery mildew resistance, and provide genetic tools for incorporating the resistance into barley breeding programmes.     

36个小麦审定和区试品种(系)的抗白粉病基因推导

明确我国当前小麦审定和区试品种含有的抗白粉病基因, 可为这些品种在小麦抗病育种中的应用及品种合理布局和轮换提供依据。本研究采用24个不同毒性的小麦白粉菌菌株对36个小麦审定和区试品种(系)进行抗白粉病基因推导, 参试品种(系)与46个已知抗病基因小麦品种(系)抗谱比较的结果表明, 11个小麦审定品种中有5个品种对所有供试白粉菌菌株表现抗性, 结合亲本溯源, 推测其中有3个品种可能携有Pm21基因; 另外6个审定品种中有5个品种可能含有抗病基因Pm2, 且其对应的亲本或亲本组合中含有抗病基因Pm2。25个区试品种(系)中有3个可能含有Pm21, 10个含有Pm2, 1个含有Pm2+6,2个含有Pm4b,1个含有Pm8。另外, 参试的36个品种(系)中还有9个品种(系)和已知基因品种抗谱存在一定差异。总体上, 推导出已知基因的品种以含有Pm2基因的品种最多, Pm21基因的品种次之, 建议在生产上加强对Pm21基因品种(系)特别是已审定的携有Pm21基因品种的推广和应用, 应该注意一些省份在育种和生产上应慎用或少用含Pm2基因的品种(系)。     

Saccharin-induced protection against powdery mildew in barley: effects on growth and phenylpropanoid metabolism

Treatment of barley ( Hordeum vulgare ) with 3 m m saccharin, applied as a foliar treatment to the first leaf or as a soil drench, provided significant control of powdery mildew ( Blumeria graminis f.sp. hordei ) on first and second leaves. This was unlikely to be the result of a direct effect of saccharin on the fungus, as application of the chemical to first leaves 2 h before inoculation did not affect conidial germination or formation of appressoria. Saccharin treatment had no significant effect on plant growth, except for a reduction in total leaf area in plants treated with a saccharin drench 14 days before inoculation with mildew. Phenylalanine ammonia-lyase activity was reduced significantly in second leaves 18 and 48 h after inoculation in plants treated with saccharin 14 days earlier. Peroxidase activity increased significantly in plants challenged with mildew within 6 days of saccharin application, although changes were not apparent until 48 h after pathogen challenge. On these occasions, treatment with saccharin resulted in a 33% increase in peroxidase activity compared with controls. In plants inoculated 10 or 14 days after saccharin application, cinnamyl alcohol dehydrogenase (CAD) activity increased prior to, and 18, 24 and 48 h after, inoculation of the barley plants with mildew. CAD activity increased approximately twofold compared with controls. However, in contrast to peroxidase, CAD activity was significantly higher in saccharin-treated plants prior to and after inoculation with powdery mildew, suggesting that saccharin primes CAD activity prior to pathogen challenge.     

燕麦种质资源抗白粉病鉴定及利用评价

采用田间自然感病的方法,于2009-2011年在甘肃省天水市甘谷县对128份燕麦品种进行了由白粉菌(Blumeria graminis f.sp.avenae)引起的燕麦白粉病田间抗性鉴定和评价,该地区属于白粉菌病害常发区。结果表明,所有供试材料均程度不同地感染燕麦白粉病,无免疫材料,2份材料‘MF9715’、‘4607’表现高抗;8份材料‘QO245-7’、‘白燕2号’、‘VAO-1’、‘709’、‘4663’、‘4641’、‘4628’和‘青永久307’表现中抗;其余118份材料表现中感、高感和极度感病。说明抗燕麦白粉病的材料严重匮乏,可利用的抗性种质资源相对更少。筛选出的高抗和中抗材料对燕麦白粉病有较好抗性,是今后可利用的抗性种质资源。     

几丁质寡糖诱导小麦抗白粉病作用的研究

试验表明 ,应用工业化生产的几丁质寡糖液 (COS)可以减轻小麦白粉病的发生。随着COS浓度的增加 ,其诱导小麦产生抗白粉病的效果越好。通过不同施用方式的比较 ,表明小麦的抗性是由于植株吸收COS后 ,引发体内系统诱导抗性而表达的。     

白粉菌侵染后田间小麦叶绿素含量与冠层光谱反射率的关系

 本文对白粉菌侵染后田间小麦叶片叶绿素含量与光谱反射率的关系进行了研究。用不同浓度的药液控制田间病害的发生梯度,于灌浆期对不同严重度小麦冠层的光谱反射率进行了测量,同时测定小麦叶绿素含量并调查病情指数,分析叶绿素含量与病情指数、冠层光谱反射率的关系。结果表明,药剂对叶绿素含量无显著影响;受白粉病危害后,小麦叶片叶绿素含量明显降低,抗性不同的品种之间下降速度存在差异,与病情指数之间也有极显著的相关性。绿光波段与叶绿素含量之间的相关性达到了显著水平;基于多波段建立的模型要优于单波段,可以利用绿光、红光波段的反射率估计叶绿素含量。红边参数与叶绿素含量之间也有较好的相关性,其中以红边面积的相关系数最大。     

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。     

不同硅源制剂对小麦白粉病的作用效果及对白粉病菌孢子萌发的影响

 研究了不同形态硅源制剂二氧化硅纳米粒子溶胶(60nm和100nm)、正硅酸乙酯、硅酸钠对小麦白粉病的作用效果以及对白粉病菌孢子萌发的影响。结果表明,在相同浓度下几种硅源制剂处理盆栽小麦幼苗对白粉病防效分别为48.78%、43.73%、40.31%和33.29%,室内处理小麦离体叶段对白粉病抑制率分别为59.67%、53.51%、51.10%和42.22%。离体叶段接种白粉菌后1.5~2h、6~7h取样,在相差显微镜下观察白粉菌孢子萌发过程,发现这几种硅制剂对白粉菌孢子初生芽管(PGT)的形成几乎没有影响,但对附着胞(AM)的形成有较大影响,其中以二氧化硅纳米粒子溶胶对AM的影响最大。     

Identification of eight effector candidate genes involved in early aggressiveness of the barley powdery mildew fungus

The barley powdery mildew fungus, Blumeria graminis f. sp. hordei (Bgh), expresses 491 candidate for secreted effector proteins (CSEPs), identified based on presence of a predicted signal peptide and relative sequence uniqueness. Virtually nothing is known about the mechanism of action of CSEPs. The present study used host‐induced gene silencing (HIGS) to identify genes involved in early fungal aggressiveness. Screening of 22 selected CSEP‐encoding genes suggested that CSEP0007, CSEP0025, CSEP0128, CSEP0211, CSEP0247, CSEP0345, CSEP0420 and CSEP0422 are individually required for normal levels of aggressiveness of Bgh. Expression data showed that these genes were up‐regulated at initial stages of infection, suggesting their involvement in early fungal aggressiveness. Altogether, the findings expand the current knowledge of genes contributing to Bgh pathogenesis and provide the basis for future functional characterization.     

Observed and predicted changes over eight years in frequency of barley powdery mildew avirulent to spring barley in France and Denmark

Aerial populations of Blumeria graminis f.sp. hordei were studied in two French and two Danish regions from 1991 to 1999, at a time of year when only winter barley was present. A high frequency of genotypes not able to grow on the spring-sown crop of the previous growing season (denoted 'spring-avirulent') was observed in most years and regions. This frequency increased with increasing proportion of winter barley; it was highest in France and decreased in general over the 8-year period. Most of the spring-avirulent genotypes possessed the V _{a 22} virulence gene, matching a resistance that has never been present in barley cultivars grown in Europe. A hypothetical cropping system, including winter- and spring-sown crops with three resistance genes altogether, was constructed to mimic the utilization of host cultivars in the four regions. Results from a mathematical model simulating changes in the composition of the pathogen population in this system, demonstrated that selection solely due to host resistance genes, i.e. without assuming any cost of virulence, might lead to such results as those observed. The changes in frequency of spring-avirulent genotypes and the frequency of unnecessary virulence genes may be predicted from the proportion of the barley area sown with winter barley, the use of resistance genes in the cultivars, the initial composition of the pathogen population, and hitch-hiking due to gametic disequilibria.     

Derivation and testing of a model to predict selection for fungicide resistance

A mathematical model was derived to predict selection for fungicide resistance in foliar pathogens of cereal crops. The model was tested against independent data from four field experiments quantifying selection for the G143A mutation conferring resistance to a quinone outside inhibitor (QoI) fungicide in powdery mildew (Blumeria graminis f.sp. hordei) on spring barley (Hordeum vulgare). Fungicide treatments with azoxystrobin differed in the total applied dose and spray number. For each treatment, we calculated the observed selection ratio as the ratio of the frequency of the resistant strain after the last and before the first spray. The model accurately predicted the variation in observed selection ratios with total applied fungicide dose and number of sprays for three of the four experiments. Underprediction of selection ratios in one experiment was attributed to the particularly late epidemic onset in that experiment. When the equation representing epidemic development was modified to account for the late epidemic, predicted and observed selection ratios at that site were in close agreement. On a scatter plot of observed selection ratios on predicted selection ratios, for all four experiments, the 1:1 line explained 89–92% of the variance in the mean of observed selection ratios. To our knowledge, this is the first fungicide resistance model for plant pathogens to be rigorously tested against field data. The model can be used with some degree of confidence, to identify anti‐resistance treatment strategies which are likely to be effective and would justify the resources required for experimental testing.     

基于小麦白粉病菌rDNA ITS序列的PCR分子检测

 Wheat powdery mildew(Blumeria graminis f.sp.tritici) is the one of main wheat diseases in China.Based on the internal transcribed spacer(ITS) sequences of ribosome of B.graminis f.sp.tritici,three molecular primer pairs(F1/R,F2/R and F3/R) were designed to detect the fungal pathogen of wheat powdery mildew.The species specificity of these primers was confirmed.F1/R was demonstrated a higher sensitivity than the other two primer pairs,and could detect as low as 1 pg DNA of B.graminis f.sp.tritici.Furthermore,F1/R primer pair was used to detect the pathogen DNA extracted from wheat leaves showing chlorosis and typical symptoms of powdery mildew caused by artificial inoculation with B.graminis f.sp.tritici.The preliminary results demonstrated the usefulness of this primer pair and its potential applications in efficient detection of wheat powdery mildew pathogen from leaves with latent infections at early growth stages of wheat.