大麦品种对白粉病抗病性鉴定及抗源筛选

通过田间初筛和病圃复筛,对6515份国内外大麦品种进行了白粉病抗性鉴定。重点材料结合苗期、分蘖期鉴定和异地鉴定,筛选出免疫或高抗材料52份,慢白粉病抗性材料59份,并发现具有Mla、Mla6单基因和Mla Mlat、Mla Mlh Mlp多基因组合的国外已知抗病基因品种,在浙江舟山和福建莆田表现抗病。统计分析表明,二棱大麦抗病性较多棱大麦强;抗病品种发病较感病品种迟;抗性材料大多来自国外,英、德、瑞典、叙利亚、澳大利亚、墨西哥等国品种,筛出率较高。     

大麦白粉病菌遗传学研究进展

大麦白粉病是由布氏白粉菌属大麦专化型活体寄生菌Blumeria graminis f.sp.hordei（Bgh）引起的真菌病害,在全球大麦种植区普遍发生,危害日趋严重。大麦白粉病菌与寄主之间存在着“基因对基因”的关系,分化为不同的生理小种或致病型。由于病原菌基因突变、重组和流动以及寄主的选择作用,大麦Bgh种群毒性、致病型频率和分布不断发生变化。随着分子生物学技术飞速发展,应用分子标记已对30多个Bgh无毒基因位点进行了连锁作图分析,已克隆了Bgh无毒基因AVRk1和AVRa10,Bgh全基因组测序现已完成。文章综述了大麦白粉病菌的侵染循环、遗传分化及其无毒基因的定位、克隆和致病机制研究进展,并探讨了基于病原菌毒性进化和基因组解码信息获得持久控制大麦白粉病的有效手段。     

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。     

陕西省小麦抗白粉病基因及新育成小麦品种(系)抗白粉病分析

为明确已知小麦抗白粉病基因载体品种以及2005—2007年陕西省新育成小麦品种(系)的抗病性,采集陕西关中不同地区小麦白粉病菌作为菌源,分年度在人工诱发病圃中对32份已知抗白粉病基因品种及578份新育成小麦品种(系)进行鉴定。结果表明,目前在陕西省仅Pm21、Pm4a、Pm5 6、Pm2 Talent及小黑麦具有优良的抗病性;Pm4b、Pm5(Mli)、Pm13、Pm19、Pm"Era"、Pm2 Mld的抗病性较弱;其它基因抗病性较差。新育成的小麦品种(系)中绝大部分对白粉病表现感病,苗期和成株期均抗病的材料仅27份,占4.67%,成株期抗病性的46份,占7.96%,其中,簇毛麦后代材料抗病性较高。     

2004—2008年东北春麦区小麦白粉病菌种群毒性动态分析

采用全国统一的9个鉴别寄主离体叶片法鉴定侵染型和Gilmour八进制法命名小种,对2004--2008年采自东北春麦区的辽宁、黑龙江等省不同县市的298个小麦白粉病菌标样进行生理小种及毒力频率分析,并利用已知抗白粉病基因的22个小麦品种(系)对参试菌株进行毒力频率测定.结果表明:2004--2006年优势小种为生理小种15号,出现频率分别为13.7%、15.6%和20.3%;2007年优势小种为17号小种,出现频率为10.2%;2008年411号小种上升为优势小种,频率达20.6%;2004--2008年毒性基因V4b、V2+6、V4+8、V12、V16、V18、V20、V21、V22和V23毒力频率较低,在0～28.0%之间,其对应抗性基因Pm4b、Pm2+6、Pm4+8、Pm12、Pm16、Pm18、Pm20、Pm21、Pm22和Pm23抗性较强,可作为有效的抗病基因加以利用.     

西南地区小麦品种(系)抗条锈病基因多样性调查

为调查西南地区小麦条锈病抗性、抗病基因位点及其组合多样性,于2013—2014年对以西南地区为主的140份小麦品种(系)进行了成株期、苗期抗病性鉴定和抗病基因标记扫描。成株期鉴定结果显示,2013年贵阳、赫章试验点和2014年贵阳、绵阳试验点都表现为抗病的品种(系)共有50份,其中表现为全生育期抗性的有37份,表现为成株期抗性的有13份;5个抗病基因Yr9、Yr10、Yr15、Yr18、Yr26的分子标记检测结果显示,西南地区小麦Yr26的使用频率最高,为41.4%,Yr9次之为37.9%,Yr10、Yr15、Yr18使用频率较低;抗条锈病基因的组合分析显示,共出现16份基因聚合品种、7种组合类型,其中组合Yr9+Yr26出现频率较高,为5%。表明西南地区的小麦品种(系)以利用全生育期抗性为主,且抗条锈病基因利用较为单一,应发掘和利用新抗条锈病基因及重视多基因的聚合。     

小麦白粉菌群体的毒性基因研究

 1987-1989年间,自四川收集的85个小麦白粉菌株接种在11个已知抗白粉病单基因小麦品种及近等基因系上。试验结果表明:v1,v3b,v3c,v5毒性基因频率很高,达90%以上,次为v3a,v7,v8(88.2-65.9%),v2,v6分别为42.4%和37.6%,v4a,v4b频率最低(15%以下),毒性基因组合中,以v2+v6,v2+v4a,v2+v4b,V4b+v6,v4a+v8的出现极少,表示含有相对应抗性基因组合的品种是高度抗病的。供试的四川小麦白粉菌株,多数均具有7-8个已知毒性基因,毒谱较广。这给该地区小麦抗白粉病育种工作增加了难度。     

大麦条纹病菌致病性分析及抗性种质精细鉴定

为探明大麦条纹病菌Pyrenophora graminea的致病性差异,鉴定筛选抗病大麦种质材料,将15株大麦条纹病菌菌株通过三明治法种子幼芽接种至5个鉴别品种,分析菌株的致病性分化情况;通过在田间孢子喷雾穗部接种和三明治法种子幼芽接种对145个田间种植表现抗病品种进行抗条纹病精细鉴定。结果显示,15株菌株对5个鉴别品种致病性有一定差异,可划分为9个致病类型,其中致病类型Ⅱ、Ⅲ和Ⅳ为优势致病类型,每个致病类型菌株数均占总菌株数的20.0%,其它致病类型的菌株均占总菌株数的6.7%;致病类型Ⅰ菌株致病力最强,对5个鉴别品种表现高感。孢子悬浮液喷雾接种田间试验结果显示,90个大麦品种表现为抗病,占总数的62.1%,其中表现免疫的品种12个、高抗品种26个、中抗品种52个,分别占总数的8.3%、17.9%、35.9%。种子幼芽接种温室条件下发病鉴定结果显示,90个抗病品种接种致病类型ⅢP05菌株,对其表现抗病的品种有28个,占供试品种总数的31.1%,表现免疫的品种10个、高抗品种7个、中抗品种11个,分别占供试品种总数的11.1%、7.8%、12.2%;接种致病类型ⅠP01菌株,对其表现抗病的品种14个,占总数的15.6%,其中表现免疫的品种4个、中抗品种10个,分别占总数的4.4%和11.1%。说明强毒性菌株P01可作为大麦条纹病抗性鉴定接种的常用菌株。     

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。     

甘肃省主要小麦生产品种及高代品系的抗白粉基因推导

［目的］ 采用基因推导法对目前甘肃省小麦主要生产品种及高代品系进行抗白粉基因分析,为甘肃省白粉病的抗病育种和品种使用及布局提供依据。［方法］ 利用21个小麦白粉菌株,结合品种系谱对甘肃14个小麦生产品种及28个高代品系进行抗白粉基因推导。［结果］ 14个生产品种中,1个含Pm8,1个含Pm4b,4个含未知抗病基因,其余8个对所有供试菌株全部表现感病;28个高代品系中,5个含Pm8〖STBZ〗,2个含Pm〖STBX〗3〖STBZ〗a,1个含Pm〖STBX〗4〖STBZ〗b,1个含Pm〖STBX〗30〖STBZ〗,5个含未知抗病基因,其余14个对所有供试菌株全部表现感病。［结论］ 目前甘肃小麦生产品种及高代品系中缺乏抗白粉病基因,一旦条件适宜,小麦白粉病在甘肃地区极易发生和流行,应该引起生产管理部门和育种专家的注意和重视。     

Virulence phenotypes of Blumeria graminis f. sp. hordei in South Africa

Virulence analysis of 224 isolates of Blumeria graminis f. sp. hordei (barley powdery mildew) from South Africa was performed. The isolates were collected from eight fields and a greenhouse in 2004 and 2007. The isolates were tested for virulence on a set of 20 differential varieties. All isolates were virulent on the resistance genes Mla8 and Ml(Ch) and avirulent for the resistance genes Mla3, Mla6, Mla7, Mla9, Mla13, Mla23, Mlp1 and MlaN81. Virulence frequencies of field isolates for the resistance genes Mla12 + MlaEm2, Mlat, Mla22, Mlk1 and Mlh were 52.9–99.5 % and for Mla1 + MlaAl2, MlLa, Mlra, Mlg + MlCP and Ml(Ru2) were 0.5–23.5 %. In total, 46 pathotypes were detected in the field and seven other pathotypes in the greenhouse. Only nine pathotypes were found in both years, but they included 61.8 % of the isolates. The predominant pathotype represented 15.9 % of the isolates, and was the only one common to all three field populations. The average relative virulence complexity per field isolate increased from 0.405 in 2004 to 0.486 in 2007. Two powdery mildew metapopulations in geographically distant and separated areas (North West and Western Cape) were deduced. The South African population of Blumeria graminis f. sp. hordei had unique virulence frequencies and virulence associations when compared to populations from other parts of the world.     

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.     

Adaptation of barley powdery mildew populations in England to varieties with two resistance genes

In 1986, two spring barley cultivars were widely grown in the UK for the first time: Klaxon, which carries the powdery mildew resistance alleles Mla7, Mlk and Ml ( La ), and Natasha ( Mla12 + Ml(Ab) ). Significant amounts of a third cultivar, Doublet ( Mla7+ Ml(La) ), were grown for the first time in 1986. The individual resistance genes, and the combination Mla7+Mlk , had previously been used separately in different varieties. Isolates of the mildew pathogen Erysiphe graminis f.sp. hordei , which were virulent on Doublet and Klaxon, were rare up to June 1986. One clone of E. g. f.sp. hordei , virulent on Doublet and Klaxon, increased in frequency from < 1% to 36% from June to October 1986, in samples from the airborne population in Cambridge, UK. This consisted of isolates with apparently identical virulences, responses to fungicides and genetic fingerprints. It also formed the majority of Doublet-virulent mildew sampled from a field of Doublet near Cambridge in 1987. By contrast, isolates virulent on Natasha were already common and genetically diverse in 1985:22 of 100 isolates sampled in October 1985, belonging to 13 races, were virulent. Natasha appeared not to influence the E. g. f.sp. hordei population greatly, as the frequency of Natasha-virulent isolates fell slightly, from 15·5% to 11·7% between June and October 1986. No single clone predominated in the Natasha-virulent population. These results support the view that new epidemics of barley powdery mildew in the UK arise by highly stochastic evolution of E. g. f.sp. hordei populations. They also indicate that varieties with new combinations of previously exposed resistance genes do not necessarily provide durable control of mildew, because the frequency of a virulent clone may rise rapidly.     

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.     

Powdery mildew resistance in selections from moroccan barley landraces

Thirty barley landraces collected from Morocco in 1985 and 1989, and held in the Polish Gene Bank, IHAR, Radzików, Poland, were screened for resistance to powdery mildew. Fifteen tested landraces (50%) showed powdery mildew resistance reactions and 24 single plant lines were selected. Eighteen lines originating from 13 landraces were tested with 17 isolates of powdery mildew and another six lines originating from six landraces were tested with 23; the isolates were chosen according to their virulence spectra observed on the ‘Pallas’ isolines differential set. Three lines (E 1090-2-2, E 1110-3-2 and E 1077-1-1) showed resistance to all powdery mildew virulence genes prevalent in Europe. In 21 lines, unknown genes alone or in combination with specific ones were detected. Five different resistance alleles(Mlat, Mlal, Mla3, Mlg andMl(CP)) were postulated to be present in the tested lines, alone or in combination:Mlat was postulated to be present in nine (~38%) lines;Mlg andMl(CP) in two lines, andMla1 andMla3 in one tested line each. The use of newly identified sources of resistance in barley breeding as a means of controlling powdery mildew is discussed.     

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.     

Virulence patterns in populations of Erysiphe graminis f.sp. hordei in Europe in 1986

A total of 3166 single colony progeny of Erysiphe graminis f.sp. hordei , constituting random samples of the aerial inoculum from 68 regions of Western Europe, were analysed in the laboratory for virulence to barley resistance genes Mlra, Mlg, Mla6+Mlg. MILa, Mla7+Mlk, Mlall, Mla9, MlaI3 + MlRu3. Mlal, Mla3 and mlo. No isolate virulent to mlo was detected. Distinct regional patterns were observed for the frequencies of most of the other virulence genes. They stressed the role of selection exerted by the cultivars grown, as well as the effect of wind dissemination of mildew populations. These two phenomena are particularly well exemplified by the data on early stages in the evolution of the newly emerging Val3 virulence in Denmark. In contrast with the preceding evolution, there was a considerable decrease of the frequency of a broadly distributed virulence ( Va6); the most probable reasons for this are outlined.     

High diversity, low spatial structure and rapid pathotype evolution in Moroccan populations of Blumeria graminis f.sp. hordei

Limited information is available about the spatial distribution and evolution of Blumeria graminis f.sp. hordei populations in North African countries, such as Morocco. Frequencies of virulence alleles in B. graminis populations are mainly driven by selection exerted by host resistance genes in addition to neutral processes such as migration and genetic drift. In Morocco, in contrast to Europe, there has been no systematic deployment of resistant cultivars, although some R genes are present in the traditional varieties. This is expected to result in the evolution of pathotypes with virulence to different R genes, and higher diversity in Morocco compared to Europe. To test this, we used 24 differential cultivars to characterise 72 isolates from Morocco in 2009. We assessed diversity and spatial structure of pathotypes and compared them to past isolates from the same area (collected in 1992). There was a high diversity of pathotypes. Isolates from 2009 were distinct from isolates from 1992, due to loss of virulence to Mla12, increased virulence to Mla8, Mla3 and Mlk1, and decreased virulence to Mla6, Ml(Ru2), Mlg and MlLa. Many virulences were different from those observed in European and Asian populations of B. graminis. At the spatial scale investigated, airborne dispersal and a lack of strong selection in the host population likely prevented the formation of population structure and allowed the accumulation of high isolate diversity. The evolution of novel and distinct pathotypes since 1992 is likely attributable to gene flow from Europe and selection by the host population in Morocco.     

Interaction between powdery mildew and barley with mlo5 mildew resistance

Powdery mildew infection of barley with the mlo5 barley powdery mildew resistance gene was examined, using near-isogenic barley lines, with and without mlo5 resistance, and two near-isogenic powdery mildew isolates, HL3/5 and GE3 with high (virulent) or low (avirulent) penetration efficiency on the resistant barley line. In all isolate–host combinations (except GE3 on the resistant barley line), frequency of haustorium formation increased significantly from zero at 11 h after inoculation to a maximum by 13 h, and there was no subsequent increase up to 24 h. In the susceptible barley line, 27% of appressoria from both isolates formed haustoria. Although this was significantly higher than the frequency of haustorium formation (18%) of HL3/5 on the resistant barley line, HL3/5 was much more successful than GE3 (frequency of haustorium formation less than 1%). The fact that HL3/5 did not possess a generally higher ability to penetrate successfully to form haustoria on the susceptible barley line, indicates that HL3/5 did not overcome the mlo5 resistance by being generally more vigorous. In the resistant barley line, papillae were larger than in the susceptible line; however, both isolates were associated with papillae of the same diameter at the time of penetration. We suggest that the mlo5 resistant barley line confers two different forms of resistance: isolate-specific and isolate-nonspecific.