Inheritance and molecular mapping of barley genes conferring resistance to wheat stripe rust

ABSTRACT Most barley cultivars are resistant to stripe rust of wheat that is caused by Puccinia striiformis f. sp. tritici. The barley cv. Steptoe is susceptible to all identified races of P. striiformis f. sp. hordei (PSH), the barley stripe rust pathogen, but is resistant to most P. striiformis f. sp. tritici races. To determine inheritance of the Steptoe resistance to P. striiformis f. sp. tritici, a cross was made between Steptoe and Russell, a barley cultivar susceptible to some P. striiformis f. sp. tritici races and all tested P. striiformis f. sp. hordei races. Seedlings of parents and F(1), BC(1), F(2), and F(3) progeny from the barley cross were tested with P. striiformis f. sp. tritici races PST-41 and PST-45 under controlled greenhouse conditions. Genetic analyses of infection type data showed that Steptoe had one dominant gene and one recessive gene (provisionally designated as RpstS1 and rpstS2, respectively) for resistance to races PST-41 and PST-45. Genomic DNA was extracted from the parents and 150 F(2) plants that were tested for rust reaction and grown for seed of F(3) lines. The infection type data and polymorphic markers identified using the resistance gene analog polymorphism (RGAP) technique were analyzed with the Mapmaker computer program to map the resistance genes. The dominant resistance gene in Steptoe for resistance to P. striiformis f. sp. tritici races was mapped on barley chromosome 4H using a linked microsatellite marker, HVM68. A linkage group for the dominant gene was constructed with 12 RGAP markers and the microsatellite marker. The results show that resistance in barley to the wheat stripe rust pathogen is qualitatively inherited. These genes might provide useful resistance against wheat stripe rust when introgressed into wheat from barley.     

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.     

天水地区不同海拔高度小麦条锈菌越冬调查初报

为进一步勘定小麦条锈病病菌越冬的海拔上限,明确不同海拔地区病害的越冬率。2008-2010年作者以甘肃省天水市甘谷县为基点,对天水地区不同海拔高度麦田小麦条锈菌的越冬情况进行了大田随机调查和病田定点系统观察。结果表明,该地区小麦秋苗发病重、越冬菌源多,是导致当地小麦条锈病春季流行的重要因素。一般年份小麦条锈菌在天水地区海拔1 293~2 080 m地带可以越冬,其越冬率随海拔降低而增高;同一地点不同年份间越冬率存在差异。越冬海拔上限与过去研究结果相比有明显提高,在海拔2 080 m地带越冬率为0.65%,可能与全球气候变暖有关。文中还对影响小麦条锈菌越冬的因素进行了讨论。     

甘肃中部麦区小麦条锈病菌越夏调查及品种抗性变异监测结果初报*

2007-2009年,对甘肃省中部麦区小麦条锈病越夏情况进行调查,结果发现：条锈病菌在定西、临夏等中部麦区海拔1 718~2 350 m地带自生麦苗上可顺利越夏,病田率和病叶率分别为63.0%~80.0%和1.05%~5.80%,其越夏菌源量比陇南更大,是当地秋苗发病的初侵染源;秋苗病情调查结果表明,中部麦区秋苗病田率和病叶率均高于同期陇南麦区;品种抗性监测结果发现,34个供试品种（系）在中部麦区和陇南麦区的抗条锈性表现基本一致,部分品种发病程度重于陇南麦区。表明甘肃省定西、临夏等中部麦区已成为陇南小麦条锈病越夏区的重要组成部分。文中还对中部麦区越夏菌源的影响、作用及品种抗性变异因素等进行了讨论。     

Genetic Diversity in Australian Populations of Puccinia graminis f. sp. avenae

ABSTRACT Sequence-tagged microsatellite profiling was used to develop 110 microsatellites for Puccinia graminis f. sp. tritici (causal agent of wheat stem rust). Low microsatellite polymorphism was exhibited among 10 pathogenically diverse P. graminis f. sp. tritici isolates collected from Australian cereal growing regions over a period of at least 70 years, with two polymorphic loci detected, each revealing two alleles. Limited cross-species amplification was observed for the wheat rust pathogens, P. triticina (leaf rust) and P. striiformis f. sp. tritici (stripe rust). However, very high transferability was revealed with P. graminis f. sp. avenae (causal agent of oat stem rust) isolates. A genetic diversity study of 47 P. graminis f. sp. avenae isolates collected from an Australia-wide survey in 1999, and a historical group of 16 isolates collected from Australian cereal growing regions from 1971 to 1996, revealed six polymorphic microsatellite loci with a total of 15 alleles. Genetic analysis revealed the presence of several clonal lineages and subpopulations in the pathogen population, and wide dispersal of identical races and genotypes throughout Australian cereal-growing regions. These findings demonstrated the dynamic population structure of this pathogen in Australia and concur with the patterns of diversity observed in pathogenicity studies.     

我国小麦条锈菌体细胞遗传重组的分子证据

 小麦条锈病是影响我国小麦生产的重要病害之一,条锈菌毒性变异是引致小麦品种抗病性丧失的主要原因。本文应用SSR分子标记技术,研究了陇南越夏区小麦条锈菌群体遗传结构,以期寻找条锈菌群体遗传重组的分子证据。研究结果表明,陇南地区小麦条锈菌群体遗传多样性比较丰富而遗传分化较小,遗传变异主要存在于群体内部,而在群体之间,遗传多样性有显著的差异。在11对SSR引物中,有4对能够检测到陇南地区小麦条锈菌群体存在遗传重组,而且重组体出现的频率不同,CPS15揭示的条锈菌重组体出现的频率为20.0%,CPS34揭示的条锈菌重组体出现的频率为18.5%,RJ20揭示的条锈菌重组体出现的频率为12.8%,RJ18揭示的条锈菌重组体出现的频率为15.0%,陇南地区小麦条锈菌群体重组体出现频率平均为16.6%。本文揭示的遗传重组现象表明陇南地区条锈菌体细胞结合十分普遍,由此推测我国小麦条锈菌在自然条件下通过遗传重组而导致毒性变异的可能性。     

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.     

中国小麦条锈菌鉴别寄主维尔对条锈病的温敏抗性研究

维尔是中国小麦条锈菌重要鉴别寄主,通过潜育期变温处理对其进行温敏抗性检测和基因推导分析。本研究发现,维尔对小麦条锈菌流行小种条中32号的抗性表现为侵染型由常温下的高侵染型向高温下的低侵染型转变,侵染指数降低,呈现高温诱导抗条锈性;通过与已知的微效基因品系比较及方差分析,推测维尔对条中32号的温敏抗性可能是由2对高温诱导表达的温敏微效基因控制。建议将维尔作为温敏微效基因资源在抗病育种中利用,作为鉴别寄主用于小种鉴定时,要严格控制鉴定温度,不要超过18 ℃。     

Genetic analysis and molecular mapping of wheat genes conferring resistance to the wheat stripe rust and barley stripe rust pathogens

ABSTRACT Stripe rust is one of the most important diseases of wheat and barley worldwide. On wheat it is caused by Puccinia striiformis f. sp. tritici and on barley by P. striiformis f. sp. hordei Most wheat genotypes are resistant to P. striiformis f. sp. hordei and most barley genotypes are resistant to P. striiformis f. sp. tritici. To determine the genetics of resistance in wheat to P. striiformis f. sp. hordei, crosses were made between wheat genotypes Lemhi (resistant to P. striiformis f. sp. hordei) and PI 478214 (susceptible to P. striiformis f. sp. hordei). The greenhouse seedling test of 150 F(2) progeny from the Lemhi x PI 478214 cross, inoculated with race PSH-14 of P. striiformis f. sp. hordei, indicated that Lemhi has a dominant resistance gene. The single dominant gene was confirmed by testing seedlings of the F(1), BC(1) to the two parents, and 150 F(3) lines from the F(2) plants with the same race. The tests of the F(1), BC(1), and F(3) progeny with race PSH-48 of P. striiformis f. sp. hordei and PST-21 of P. striiformis f. sp. tritici also showed a dominant gene for resistance to these races. Cosegregation analyses of the F(3) data from the tests with the two races of P. striiformis f. sp. hordei and one race of P. striiformis f. sp. tritici suggested that the same gene conferred the resistance to both races of P. striiformis f. sp. hordei, and this gene was different but closely linked to Yr21, a previously reported gene in Lemhi conferring resistance to race PST-21 of P. striiformis f. sp. tritici. A linkage group consisting of 11 resistance gene analog polymorphism (RGAP) markers was established for the genes. The gene was confirmed to be on chromosome 1B by amplification of a set of nullitetrasomic Chinese Spring lines with an RGAP marker linked in repulsion with the resistance allele. The genetic information obtained from this study is useful in understanding interactions between inappropriate hosts and pathogens. The gene identified in Lemhi for resistance to P. striiformis f. sp. hordei should provide resistance to barley stripe rust when introgressed into barley cultivars.     

冀鲁豫部分小麦生产品种在甘肃陇南的抗条锈性表现

2007-2008年,在甘肃省农科院植保所甘谷试验站及天水市的不同海拔高度地区,对收集到的146份冀鲁豫生产品种进行了全生育期抗条锈性鉴定及成株期抗条锈性监测。结果发现,仅有绵麦39抗性表现稳定,对接种鉴定及自然诱发的条锈病均表现免疫,合选18、许6、矮抗58等3份材料的抗性表现也相对较好,抗性比例仅为2.74%,其余142份材料均表现感病。文中还对各品种利用前景、评价等进行了讨论。     

欧洲小麦品种Mega抗条锈病基因的遗传分析及分子标记

 本研究表明欧洲小麦品种Mega对我国小麦条锈病重要流行小种CYR30、CYR31、CYR32、Su-4和Su-14在苗期都具有良好的抗病性。采用小麦条锈菌小种CYR30对Mega与感病小麦品种铭贤169杂交的F₁、F₂和BC₁代及双亲进行苗期抗病性遗传分析,结果表明,Mega对CYR30的抗性由1对显性基因独立控制。采用SSR标记技术对其携带的抗性基因进行分子标记,在237对SSR引物中,发现位于5BL上的2个SSR引物位点Barc232、Wmc640在双亲和抗、感池间能扩增出稳定的特异性片段,与抗病基因连锁的遗传距离分别是3.7cM和8.6cM,暂命名为YrMe。本研究结果为科学利用Mega抗条锈基因培育抗病品种提供了依据。     

86份贵协系小麦种质资源对条锈病的抗病性评价

小麦条锈病是发生于甘肃陇南麦区小麦生产上的最主要病害,利用和种植抗病品种(系)是防治该病害最经济有效且有利于保护环境的措施。2011-2013年度连续两年在甘肃省农业科学院植物保护研究所甘谷试验站田间对86份来自贵州大学的贵协系小麦种质资源材料进行了成株期抗条锈病鉴定,结果发现仅有‘贵协1’、‘贵协2’、‘贵协3’、‘贵协7’、‘贵协011-2-6’、‘贵协011-3-16’、‘贵协011-3-29’、‘贵协011-3-31’、‘贵协011-3-34’等9份材料表现抗病,且抗病性相对稳定,可作为一线抗源材料在陇南抗病育种中充分利用。     

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。     

应用real-time PCR 定量检测田间小麦条锈菌潜伏侵染的研究

 为快速、及时地定量检测处于潜育状态下的小麦条锈菌菌量,本研究于2009－2010年实施田间试验,在北京和甘肃各设3个试验小区,将小区分割成1 m×1 m或1.5 m×1.5 m的小格。初春刚返青叶片出现症状前,在每小格内采集30片叶,提取叶片基因组DNA,应用已建立的real-time PCR定量测定方法,计算每个样本的分子病情指数(molecular-detected disease index, MDI）。在本试验的天气条件和试验设计下,采样10 d后,调查田间标记采样点的发病情况,计算病情指数DI,比较MDI和DI的关系,建立回归模型。结果表明：甘肃甘谷3个试验小区的MDI和DI存在极显著的相关性（R₂¹=0.813 6,R₂²=0.884 5,R₂³=0.592 8）;北京上庄3个试验小区的MDI和DI也存在极显著的相关性（R₂¹=0.742 3,R₂²=0.578 4,R₂³=0.858 4）。本研究证明了应用分子生物学方法可估测小麦条锈病潜育侵染程度的可能性,为建立分子定量测定田间潜育侵染量系统提供了可靠依据。     

Models for predicting potential yield loss of wheat caused by stripe rust in the U.S. Pacific Northwest

Climatic variation in the U.S. Pacific Northwest (PNW) affects epidemics of wheat stripe rust caused by Puccinia striiformis f. sp. tritici. Previous models only estimated disease severity at the flowering stage, which may not predict the actual yield loss. To identify weather factors correlated to stripe rust epidemics and develop models for predicting potential yield loss, correlation and regression analyses were conducted using weather parameters and historical yield loss data from 1993 to 2007 for winter wheat and 1995 to 2007 for spring wheat. Among 1,376 weather variables, 54 were correlated to yield loss of winter wheat and 18 to yield loss of spring wheat. Among the seasons, winter temperature variables were more highly correlated to wheat yield loss than the other seasons. The sum of daily temperatures and accumulated negative degree days of February were more highly correlated to winter wheat yield loss than the other monthly winter variables. In addition, the number of winter rainfall days was found correlated with yield loss. Six yield loss models were selected for each of winter and spring wheats based on their better correlation coefficients, time of weather data availability during the crop season, and better performance in validation tests. Compared with previous models, the new system of using a series of the selected models has advantages that should make it more suitable for forecasting and managing stripe rust in the major wheat growing areas in the U.S. PNW, where the weather conditions have become more favorable to stripe rust.     

81份春小麦种质资源抗条锈病评价

本研究结合苗期分小种接种和大田成株期自然诱发条件下对供试的81份春小麦资源进行抗条锈病鉴定,分析其抗条锈病情况、抗病类型及抗病性关系,结果表明:具全生育期抗条锈病类型的资源有7份,具成株期抗条锈病类型的资源有39份,有30份资源为含有成株期和对部分小种失去抗性的全生育期抗病性的抗性类型;苗期分别接种7个小种后,表现抗病的资源数为9~25份不等,表现中抗的资源为2~6份不等,表现感病的有50~69份不等,成株期表现抗病的73份,表现中抗的3份,感病的5份,即供试种质资源大部分表现为苗期感病,而成株期具有抗病性。聚类分析结果表明,该81份种质资源抗性聚类相对分散,遗传多样性水平较高。研究结果为小麦育种提供优良的抗条锈性资源。     

印度圆粒小麦抗条锈病新基因YrSph的遗传分析和SSR标记定位

 抗条锈病小麦新种质D31是以印度圆粒小麦（Triticum sphaerococcum Perc.）AS384与普通小麦品系94-3854杂交、回交选育而成的新品系。用中国小麦条锈病菌（Puccinia striiformis f.sp. stritici）流行生理小种条中32号对D31和Taichung29的杂交后代进行苗期及成株期抗条锈性遗传分析。结果表明,D31对生理小种条中32号表现为全生育期近免疫抗性反应,其抗性由1对隐性基因控制,暂命名为YrSph。利用BSA法对构建的F²遗传作图群体进行SSR标记分析。通过对400对微卫星引物的筛选和群体分析表明,抗性基因与 Xwmc149、Xwmc246、Xgwm372和Xwmc198 具有连锁关系,其中与 Xwmc246 标记连锁较近,遗传距离为8.8 cM。基因和标记之间的顺序为 Xwmc149-YrSph-Xwmc246-Xgwm372-Xwmc198 。根据作图结果,将D31所含的抗条锈病基因YrSph定位于2A短臂上。基于该基因的作图位置与系谱分析,认为该基因可能是1个新的抗条锈病基因。     

四川省100个小麦品种(系)抗条锈病基因的 分子检测

 为了解四川省近年小麦生产品种和后备品种对条锈菌致病类型G22-83和流行小种CYR32、CYR33的抗性水平,明确已知的主要抗条锈基因在该地区小麦品种中分布状况,利用小麦条锈菌G22-83、CYR32和CYR33对四川省100个小麦品种(系)进行苗期抗病性鉴定;采用已知基因Yr5、Yr9、Yr10、Yr15、Yr18 和Yr26的分子标记,对供试小麦材料进行抗性基因检测。结果表明,100个供试材料中,对CYR32 表现抗病的58份,占58%;对CYR33 表现抗病的63份,占63%;对G22表现抗病的43份,占43%;对CYR32、CYR33和G22均表现抗病性的品种(系)28个,占28%。供试小麦品种(系)中携带Yr9、Yr10、Yr15 和Yr26基因的材料分别有24份(24%)、9份(9%)、5份(5%)和26份(26%),所有供试品种(系)中未检测到Yr5和Yr18基因。     

BTH和INA诱导小麦抗条锈病性的研究

用不同浓度的苯(1,2,3)噻二嗪-7-硫代羧酸-S-甲酯(BTH)或2,6-二氯异烟酸(INA)喷雾处理小麦幼苗,进行小麦诱导抗病性研究。结果表明,以1.0mmol(毫摩尔)INA处理后接种小麦条锈病的诱导抗性效果最好。以1.0mmolINA和0.3mmolBTH处理小麦叶片5d后接种,叶片中苯丙氨酸解氨酶(PAL)和多酚氧化酶(PPO)的活性均有明显上升,且PAL、PPO和INA、BTH的诱导抗性表达都呈正相关关系。BTH和INA水溶液对小麦条锈菌夏孢子的萌发均无明显抑制作用。据此认为,处理后病情指数的下降是由于试剂处理提高了小麦幼苗的抗病性,即麦苗产生了诱导抗性而引起的。     

条锈菌诱导的小麦EF手钙离子绑定蛋白基因TaCab1的功能初步分析

 根据小麦EF手钙离子绑定蛋白(TaCab1)基因序列,利用WMD3软件设计特异的人工miRNA (amiRNA),构建VIGS沉默载体。利用amiRNA-VIGS体系,对小麦的TaCab1基因的功能进行了初步分析。利用Northern blot和实时定量PCR技术分别检测了amiRNA的积累及TaCab1的沉默效率,并利用显微观察技术统计条锈菌侵染小麦后的组织学差异。结果表明,amiRNA可以得到有效的积累,其靶标基因TaCab1可以得到有效的沉默。从表型上看,小麦叶片上条锈菌夏孢子的产孢量也在一定程度上有所降低。组织学观察发现当TaCab1被沉默后,寄主细胞的坏死面积在侵染后期明显增大,条锈菌的菌丝分枝数也明显增多,但菌丝长度明显变短。