Temperature adaptation in Australasian populations of Puccinia striiformis f. sp. tritici

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the major fungal pathogens of wheat. A new pathotype was introduced to Australia in 2002 and several derivative pathotypes were detected in subsequent seasons. It has been suggested that the severity of stripe rust outbreaks in Australia since 2002 could be as a result of traits other than virulence in the pathogen population. This study was conducted to investigate the hypothesis that the stripe rust pathogen population dominant in Australia since 2002 was better adapted to warm temperature conditions compared to previous pathogen populations. Sixteen pathotypes were selected to examine the influence of two contrasting temperature regimes during the 24 h incubation (10°C and 15°C) and the subsequent post‐inoculation (17°C and 23°C) periods on latent period and infection efficiency on four susceptible wheat cultivars. In addition, the effect of two contrasting incubation temperatures on urediniospore germination was examined. The results indicated that pathotypes of P. striiformis f. sp. tritici detected after 2002 did not show evidence of adaptation to high temperatures, which suggests that other factors contributed to the observed increased aggressiveness.     

Thermal generalist behaviour of invasive Puccinia striiformis f. sp. tritici strains under current and future climate conditions

Yellow rust is a devastating wheat disease. Since 2000, Puccinia striiformis f. sp. tritici strains PstS1 and PstS2 have become adapted to high temperatures and have spread worldwide. By 2011, Warrior strains had invaded both warm and cold areas of Europe. This study questioned whether thermal aptitude promoted the spread of Warrior strains, similar to PstS1/PstS2, by comparing infection efficiency (IE) at five temperatures and latent period (LP) under warm and cold regimes for Warrior isolates and pre‐2011 reference strains on two susceptible wheat varieties. The Warrior isolates showed a range of IE and LP responses to temperature that was intermediate between the northern reference isolates adapted to cold conditions and both the southern and invasive PstS2 isolates adapted to warm conditions. Warrior isolates had the highest IE under optimal temperatures of 10 and 15 °C, and displayed reduced infectivity under the warmest (20 °C) and coldest (5 °C) temperatures. Warrior strains acted as thermal generalists and the reference isolates acted as specialists. An IE thermal response was used to simulate the development of each isolate under future climate scenarios in a temperate and Mediterranean region. Isolates had the same ranking for yearly IE over the three 30‐year periods (1971–2000, 2021–2050, 2071–2100) and both locations, with a slight infection increase in the future. However, in the future IEs increased in earlier months. The thermal generalist profile of Warrior isolates for IE was confirmed, with an intermediate capacity to tolerate warming climate, whereas the southern isolates are better adapted to warm conditions, but do not have the virulences necessary to develop on current varieties.     

Dry-dispersal and rain-splash of brown (Puccinia recondita f.sp. tritici) and yellow (P. striiformis) rust spores from infected wheat leaves exposed to simulated raindrops

The dispersal of spores from lesions of brown ( Puccinia recondita f.sp. tritici ) or yellow ( P. striiformis ) rusts of wheat by impacting drops was studied. Using a generator of uniform-size drops, drops of 2.5, 3.4, 4.2 and 4.9 mm in diameter were released from rest at heights of 5, 50 and 100 cm above horizontal and primary leaves uniformly covered with sporulating lesions. Dry-dispersal and rain-splash occurred simultaneously in response to drop impaction. A coloration technique allowed separate counting of dry-dispersed and rain-splashed spores caught on slides. More spores were rain-splashed than dry-dispersed. Neither removal mechanism affected in-vitro germination of spores, which was higher in brown than in yellow rust. For both rusts, the number of both dry-dispersed and rain-splashed spores, as well as their travel distance, increased with drop diameter and fall height. The fall speed of incident drops in relation to diameter and fall height was obtained by solving numerically the equation of vertical drop motion. The number of spores removed by a given impacting drop was found to be a power function of the calculated kinetic energy of the impacting drop. Based on this experimental relationship, a simulation study showed the relevance of rain type in the removal of spores.     

Genetic diversity of Puccinia striiformis from cereals in Alberta,Canada

Stripe rust of wheat caused by Puccinia striiformis f. sp. tritici has recently become a production problem on wheat in Alberta, Canada, and stripe rust of barley caused by P. striiformis f. sp. hordei occurs regularly. A total of 261 isolates of P. striiformis were collected from wheat, barley, Hordeum jubatum and triticale plants in Alberta, Canada from 2007 to 2012, and compared to isolates from other provinces and the USA. The genetic diversity of the pathogen was assessed using 11 simple sequence repeat (SSR) markers and by examining a length polymorphism in the ribosomal DNA (rDNA) intergenic spacer 1 (IGS1) region. A total of 28 SSR genotypes were detected within Alberta. The 13 genotypes common on wheat (P. striiformis f. sp. tritici) were distinct from the 15 genotypes common on barley (P. striiformis f. sp. hordei). Four SSR genotypes, two within each forma specialis, represented 85% of the isolates recovered. Genotypic diversity was low, population genetic analysis indicated a clonal structure, and the genotypes were widely dispersed. In both formae speciales, the dominant genotype varied between years. The second most common P. striiformis f. sp. hordei genotype was found to be more closely related to older P. striiformis f. sp. tritici genotypes from the USA than to other P. striiformis f. sp. hordei genotypes.     

小麦条锈菌cDNA文库构建和表达序列标签(ESTs)分析

 小麦条锈菌(Puccinia striiformis f. sp. tritici)是全世界范围内小麦生产上的重要病原真菌, 但是对小麦条锈菌的基因组和基因功能却了解甚少。为了促进小麦条锈菌基因组学的发展和大规模基因发现, 我们以噬菌体λTrip1Ex2为载体, 采用SMART技术构建了小麦条锈菌萌发夏孢子的cDNA文库。原始文库的滴度为1.1×10⁶pfu/mL, 平均插入片段长度为750 bp。从文库中随机挑取279个cDNA克隆测序, 分析发现这些ESTs的平均GC含量为45.08%。通过聚类分析, 279个ESTs拼接成31个contigs和80 singletons。BLASTx分析表明, 47%的ESTs与GenBank中报道的功能已知或未知蛋白具有相似性。tBLASTx分析表明12个uniseqs与EST数据库中的序列具有相似性, 其中9个是来自担子菌的cDNA文库。几个EST与已知的真菌致病相关基因具有高度相似性。RT-PCR分析了几个基因在小麦条锈菌侵染过程中的表达水平。这些结果为小麦条锈菌夏孢子萌发以及侵染寄主过程中的基因表达研究奠定了很好的分子生物学基础。     

不同培养载体对小麦条锈菌夏孢子人工萌发的影响

为探索适合小麦条锈菌寄主体外萌发的载体,并建立基于不同研究目的的小麦条锈菌夏孢子人工萌发技术,采用超声波水雾培养法,以小麦条锈菌生理小种CYR32为供试材料,探讨了不同培养载体对小麦条锈菌夏孢子萌发特性的影响.在最适温度和湿度条件下,以4种不同材料为培养载体,小麦条锈菌夏孢子的萌发率、芽管生长率和外观显示情况有显著差异.在尼龙纱网上的夏孢子萌发速度和芽管伸长速度最快,12 h内萌发率最高可达93.40％,芽管最长可达446.71 μm;亲水滤膜上夏孢子的萌发率和芽管长仅次于纱网;亲水塑料膜和PVDF膜上夏孢子的萌发率均高于普通亲水载玻片,而芽管长度低于普通亲水载玻片.采用侧光照明时,在亲水滤膜和PVDF膜上极易清晰观测到夏孢子及其芽管内的黄色颗粒物.在亲水塑料膜上,还可观察到夏孢子萌发12 h后,芽管尾部膨大形成类似附着胞的结构.结果显示,超声波水雾可使条锈菌夏孢子处于水分饱和的环境,而纱网纤维组成的小格具有很好的保水结露功能,能使夏孢子萌发快,萌发效率高,芽管生长快;另一方面,使用其他培养载体还可满足观测条锈菌夏孢子内黄色颗粒物向芽管内转移和芽管尾部形成类似附着胞结构的目的.     

利用小种标记分析云南省两个县的小麦条锈菌群体

 小麦条锈病是由小麦条锈菌（Puccinia striiformis f.sp. tritici）引起的气流传播病害,在世界各主要麦区均有发生,也是我国小麦生产上危害最严重的病害之一。小麦条锈菌毒性变异频繁,易产生新毒性小种,导致小麦品种的抗病基因失效,引起小麦条锈病周期性流行。因此,了解小麦种植区的条锈菌群体结构组成及变异,对于制定更为有效的条锈病控制方法具有重要的意义。     

云南省德宏州小麦条锈菌越夏调查初报

小麦条锈菌越夏菌源量直接影响着小麦条锈病的发生与流行。为了明确云南省德宏州是否是小麦条锈病菌的越夏菌源地,2010年和2011年在德宏州芒市江东乡海拔1 600、1 700、1 800、1 900m和2 000m处设置了小麦条锈病菌源观察圃,对观测圃中小麦品种和周围落粒自生麦苗上条锈病的发病情况进行调查,实时记录各观察点的温湿度。结果表明:(1)海拔2 000m的菌源观察圃能观测到小麦条锈菌越夏和越冬的情况,越冬菌源量和春季条锈病的发病情况存在相互影响的关系;(2)海拔1 900m及以上的地区最高日平均温度和旬平均温度均低于22℃,为小麦条锈菌的越夏区域;1 600~1 900m的区域内其最高日平均温度在22℃至25℃之间,最高旬平均温度均大于20℃小于23℃,符合小麦条锈菌越夏的理论温度要求;并且海拔1 900m及以上地区的菌源观察圃中可观察到小麦条锈菌越夏。以上结论证实了云南省德宏州芒市是小麦条锈菌的越夏菌源地之一。     

中国小麦条锈菌主要流行菌系的寄生适合度研究

 为了预测我国未来小麦条锈菌的优势种群,本研究采用抗性组分法测定了我国条锈菌主要流行菌系的寄生适合度。结果表明:决定我国小麦条锈菌最重要的寄生适合度属性为条锈菌的产孢能力(繁殖力),其次为条锈菌的侵染能力,再次为扩展能力;建立了小麦条锈菌寄生适合度属性3个主成份的数学模型;在供试的菌系中,Su11-4、Su11-14和CY32相对寄生适合度较高,已经成为我国目前小麦条锈菌的优势小种(类型);CY31由于其寄生适合度较低,出现频率逐年下降,已成为次要小种,感染"中四"苗期的新菌系T4类型,由于其寄生适合度低,也不会成为今后一个时期的流行小种。     

中国小麦条锈菌4个主要流行小种的寄生适合度

 以12个全国大面积种植的小麦生产品种和1个高感品种为试材,利用抗性组分法研究了中国小麦条锈菌(Puccinia striiformis f.sp.tritici,Pst)4个主要流行小种条中32号、条中31号、水源11-14和水源11-4的寄生适合度。因子分析结果表明,病害量是度量条锈菌生理小种寄生适合度最重要的参数,其次是夏孢子生活力、产孢能力、产孢面积、产孢期,由此提出寄生适合度=(产孢量×侵染概率×夏孢子堆密度×夏孢子萌发率)/潜育期。研究还发现,条中32号的寄生适合度最高,致病性最强,与目前该小种为优势小种的事实相符;水源11-14的寄生适合度较高,有较大的发展潜力,在抗病育种中应予以高度重视。     

Clonality and long-distance migration of Puccinia striiformis f.sp. tritici in north-west Europe

The biotrophic fungus Puccinia striiformis f.sp. tritici , a basidiomycete that causes yellow rust on wheat, is spread by wind-dispersed spores. Analysis of amplified fragment length polymorphism (AFLP) variation showed that the fungus frequently migrates between the UK, Germany, France and Denmark. There is no biological evidence for sexual or parasexual reproduction under natural conditions, and this was supported by the lack of recombination, as revealed by AFLP, over the time and area represented by the samples in this study. A phylogeographic analysis revealed that there was effectively a single, clonal population in the four countries, up to 1700 km apart, consistent with a 'continent-island' model in which Denmark is the recipient of migrants from other countries. In five cases, specific pathogen clones were dispersed between the UK and Denmark, and on at least two recent occasions clones were also spread from the UK to Germany and France, causing outbreaks of yellow rust on wheat cultivars that were previously resistant to the disease in these countries. The agronomic consequences of migration were enhanced because of the limited genetic diversity for yellow rust resistance in wheat cultivars in the area. These results demonstrate that long-distance migration of pathogen clones, coupled with low diversity in the host species, may cause previously useful resistance genes to become ineffective for disease control on a continental scale.     

2010年西藏小麦条锈菌生理小种群体结构与分析

西藏地区是中国相对独立的小麦种植区,小麦条锈病是西藏冬小麦上最重要的病害.长期以来,对西藏小麦条锈菌生理小种群体结构缺乏全面系统的了解.为了弄清西藏小麦条锈菌生理小种群体结构,本研究从西藏地区小麦条锈病发生的关键地区采集并鉴定了小麦条锈菌标样261份.西藏地区小麦条锈菌群体结构复杂,小种类型数多,主要优势小种以CYR32和CYR33为主,水源11类群为优势类群,Hybrid46类群结构简单,未发现CYR32以外的类型;CYR32之前的小种数较多、其中CYR17、CYR20、CYR31出现频率较高;西藏小麦条锈菌群体结构与内地有着较大的相似性,同时也有其自身的独特性,表现西藏小麦条锈菌优势小种组成与四川、云南两省相似,与青海省差异较大.推测西藏地区小麦条锈菌与四川和云南省存在较密切的菌源交流,与青海省交流较少.     

我国小麦秋苗条锈病发生规律及其区间菌源传播关系

全国冬小麦秋苗均可遭受来自西北和西南越夏区条锈菌的侵染与危害, 条锈菌在寄主上进一步繁殖和发展, 引起本地或者外地小麦条锈病流行。不同麦区秋苗条锈病发生时间、发生程度及其菌源传播规律各不相同, 年度间亦存在差异。根据小麦条锈病发生流行频率、病菌越冬和越夏情况、秋季菌源和春季菌源的有无与多少、提供时间及其影响范围与作用, 结合地理生态条件、气候特点、小麦种植区划与栽培模式等, 将中国小麦条锈病发生流行区域划分为8个明显不同的生态区系, 即关中、华北春季流行区; 成都平原、江汉流域冬季繁殖区; 西北、川西北越夏易变区; 云贵高原越夏冬繁区; 新疆冬春麦常发区; 西藏高原青稞、小麦常发区; 南方晚播冬麦偶发区; 内蒙古、东北春麦偶发区。通过病害实地调查、病菌群体遗传多样性和高空气流轨迹分析, 进一步揭示了区间菌源传播关系。陇南、陇中、陇东、宁南、海东、陕西宝鸡以及川西北和云贵高原等地区离越夏区较近, 冬小麦播种较早, 秋苗条锈病发生早、发病重, 秋季随西北气流传播到平原冬麦区和海拔较低的冬麦区侵染危害秋播麦苗, 其菌源数量对全国小麦条锈病发生流行程度起着至关重要的作用, 是中国小麦条锈病的秋季菌源基地, 面积约67万hm2; 成都平原、江汉流域、陕南、豫南、云贵坝区等麦区, 离条锈菌越夏区相对较远, 小麦播种期也较晚, 秋苗发病较晚较轻, 但冬季气候温和, 雨露条件充沛, 条锈菌在冬季可以不断侵染和繁殖, 在早春可积累大量菌源, 然后向北部和西部广大麦区扩散传播, 引起小麦条锈菌春季侵染, 是中国小麦条锈病的春季菌源基地, 面积约200万hm2。云贵高原越夏冬繁区可为我国广大麦区特别是长江中下游麦区提供部分菌源, 荆州等鄂东南地区是云贵菌源向长江下游麦区传播的中转站。云南与甘肃之间存在大量的基因流, 推测云南可能是中国小麦条锈菌重要的起源中心。西南和西北秋季菌源对长江流域麦区的相对重要性有待进一步研究明确。     

应用HYSPLIT-4模式分析小麦条锈病菌远程传播事例

 小麦条锈病的发生及大区流行与病原菌的远程传播密切相关。预测小麦条锈病大区流行的关键是搞清病原菌的远程传播路线和孢子云浓度变化规律。气流是菌源传播的主要动力,因此,气流运动的物理模型是对小麦条锈病菌远程传播事例分析的有力工具。本研究利用再分析格点资料(NCEP/NCAR Reanalysis Data),基于HYSPLIT-4模式,对历史上我国小麦条锈病菌远程传播的经典事例(分别发生于1960、1964、1975和1983年)进行了分析,着落区孢子浓度时间序列模拟结果与观察到病害发生前几天的降雨事件是相符的,结果表明小麦条锈病的远程传播及发生时间可通过计算大气环流运动来预测,病原菌孢子的沉降除由于孢子自身重力而引起的沉降外,很大程度上还受到降雨引起的湿沉降的影响。HYS-PLIT-4模式可用于探讨我国小麦条锈病区域间菌源传播关系问题,进行病害的预测预报,进一步指导我国小麦条锈病越夏和越冬调查以及菌源区综合治理和小麦品种布局。该研究在方法和技术上也可为研究其它病原菌远程传播提供借鉴。     

Attempted somatic hybridization of Puccinia striiformis f. sp. tritici and P. striiformis f. sp. hordei

Attempts were made to produce somatic hybrids between isolates of Puccinia striiformis f. sp. tritici and f. sp. hordei. A mixed infection was produced on a common susceptible barley host, Fong Tien, using white-spored isolates of P. striiformis f. sp. tritici and yellow-spored isolates off. sp. hordei. Selection was made for non-parental spore colour on selective wheat and barley hosts, and variants thus isolated were analysed for virulence markers, and for isozyme and double-stranded RNA (dsRNA) markers, all of which clearly differentiated the parental isolates. Two white-spored (non-parental) isolates were found on the selective barley host which otherwise resembled the parental f. sp. hordei isolate in virulence, isozyme and dsRNA markers. The most likely explanation of the origin of these isolates is mutation to white spore colour in the f. sp. hordei isolate.     

新疆伊犁州小麦条锈菌生理小种鉴定及毒性分析

伊犁州是新疆小麦条锈病重灾区, 对该地区开展小麦条锈菌生理小种监测意义重大?本研究通过对2020年采自新疆伊犁4县的149份小麦条锈菌样品进行生理小种监测, 以期明确该地区小麦条锈菌生理小种组成及毒性情况?结果表明, 共监测到28个生理小种, 其中CYR 33?Su 11-1?Su 11-12?CYR 32及CYR 34出现频率较高, 分别为14.09%, 12.75%, 8.05%, 8.05%和7.38%; 水源11类群为优势类群, 出现频率高达44.30%?对抗条锈病基因Yr1?YrA?Yr3?Yr6?YrSu?Yr9的毒性频率均大于70%, 表明这些基因在伊犁州抗性基本丧失?新疆伊犁州4县小麦条锈菌毒性多样性分析显示, Nei’s 遗传多样性指数为0.34, Shannon 信息指数为0.50, 表明伊犁州条锈菌毒性多样性水平较高, 毒性组成丰富; 小麦条锈菌毒性相似系数在0.92~1.00, 其中伊宁县和巩留县的样品遗传距离最近, 察布查尔县与其他3县样品遗传距离最远?因此, 新疆伊犁州地区抗锈育种应以抗CYR 33和Su 11-1为主, 兼顾抗Su 11-12?CYR 32和贵农22类群中其他类型?另外不同县区应合理进行抗病基因布局, 以期实现小麦条锈病的区域间联合防治?     

2017年2018年甘肃省小麦条锈菌生理小种变异监测

2017年2018年对采自甘肃省不同生态区的572份小麦条锈菌标样进行生理小种鉴定及监测, 结果表明:两年共监测出37个生理小种及致病类型。其中贵农22类群居第一位, 2017年和2018年出现频率分别为56.88%和50.34%, 中四类群出现频率分别为10.15%和13.85%。主要流行小种中, 条中34号出现频率分别为33.69%和38.51%, 居第一位; 条中32号出现频率分别为13.04%和15.20%, 居第二位; 条中33号出现频率降至5%以下; 贵22-14致病类型出现频率在4%~6.6%之间; 新菌系ZS-1出现频率呈上升趋势, 2017年和2018年出现频率分别为2.90%和7.43%; 次要致病类型中, HY4、HY8、水11-3、水11-7、水11-192及贵22-104、贵22-108、ZS-18等出现频率在0.72%~4.71%之间, 其他致病类型在0.36%以下。毒性分析发现, VYr9、VYr3b+Yr4b、VYrSu、VYr26仍为甘肃省主要毒性致病基因。目前甘肃小麦条锈菌仍处于以条中34号为主的贵农22致病类群占优势阶段。开展中四类群监测是今后条锈菌变异研究的重点。抗条锈育种应以抗条中34号、条中32号、ZS-1为主, 兼顾贵农22及中四类群中其他类型。     

2010-2011年小麦条锈菌群体的温度敏感性与毒性及遗传多样性的关系

为了明确小麦条锈菌毒性、遗传多样性以及温度敏感性三者之间的关系,本研究利用21个已知抗条锈病基因近等基因品系对2010-2011年生长季采自6个省市78株已知温度敏感性(ET50)的小麦条锈菌群体进行毒性鉴定,并利用AFLP技术对其进行遗传多样性分析。苗期毒性鉴定结果表明,不同省市小麦条锈菌群体的毒性基因多样性存在一定差异,甘肃菌株群体毒性多样性指数H值最高,为0.269 3,云南菌株群体最低,为0.150 4。遗传多样性结果显示,6省市小麦条锈菌群体遗传多样性指数H值范围在0.125 5~0.165 3之间,遗传一致度GI为0.964 7~0.987 2,遗传距离GD为0.012 9~0.036 0。三者的相关性分析表明,条锈菌群体毒性多样性与平均ET50显著负相关,与ET50变异系数正相关,而与遗传多样性没有显著的相关关系。     

2019年-2020年甘肃省小麦条锈菌生理小种变异监测

2019年-2020年, 对甘肃省不同生态区于小麦不同生育期采集的661份条锈菌标样, 在甘肃省农业科学院植物保护研究所兰州温室进行条锈菌生理小种变异监测, 结果表明:两年共监测到52个生理小种及致病类型, 其中贵农22仍为优势致病类群, 居第一位, 2019年和2020年出现频率分别为36.7%和 49.5%, 低于2018年的50.3%; 其次是HY和水源致病类群, 出现频率分别为25.3%?25.5%和10.2%和15.5%; 中四致病类群出现频率分别为22.8%和7.6%, 2020年居第五位, 低于2018年的13.9%?流行小种条中34号出现频率分别为23.4%和22.5%, 居第一位; 条中32号出现频率分别为19.6%和17.9%, 居第二位; 条中33号?贵22-14出现频率分别为4.4%?4.0%和2.5%?6.0%; 新类型ZS-1出现频率分别为7.0%和0.6%?次要致病类型如HY8?HY29?HY103?水11-192及贵22-13?贵22-108?贵22-244和贵22-271等出现频率在1.0%~1.9%之间, 其他致病类型均在0.6%以下?毒性分析发现, VYr9?VYr3b+Yr4b?VYrsu?VYr26为甘肃省条锈菌主要毒性致病基因, 毒性频率在50.6%~93.7%之间?甘肃小麦条锈菌群体结构仍处于以条中34号为主的贵农22致病类群占优势阶段?对感染中四的新致病菌系ZS进行持续监测是今后一段时期甘肃省小麦条锈菌变异研究的重点?小麦抗条锈育种应以兼抗条中34号?条中32号为主, 兼顾贵农22其他类型及中四新菌系?     

小麦条锈菌冬孢子生物学特性研究进展

由小麦条锈菌引起的小麦条锈病是我国广大麦区的严重病害,研究其冬孢子产生原因与作用对于解析该病菌的生活史及遗传变异机制至关重要.通过对国内外相关研究成果的整理,综述了小麦条锈菌冬孢子的形态特征、生物学特性、寄主范围及其作用与功能等方面的研究进展.