河南省西部山区小麦白粉菌群体遗传多样性分析

为揭示河南省小麦白粉菌群体遗传结构、起源及进化关系,采用简单重复序列区间(inter-simple sequence repeats,ISSR)和扩增片段长度多态性(amplified fragment length polymorphism,AFLP)分子标记技术对河南省西部山区4个小麦产区的35个小麦白粉菌单孢分离菌株进行了群体遗传多样性分析。结果显示:ISSR和AFLP分析均将35个菌株分为3个组,组Ⅰ包括来自卢氏和灵宝的大部分菌株;组Ⅱ包括来自栾川、卢氏和巩义的菌株;组Ⅲ由4个地区的个别菌株组成,同时包含1个闭囊壳释放子囊孢子获得的菌株。ISSR分析出菌株遗传距离分布在0.0139～0.6592之间,扩增多态性比率为64.83%,各菌株间的Shannon指数为0.2749;而AFLP分析所得的各菌株遗传距离变化幅度在0.1257～0.9322之间,扩增多态性比率为82.68%,各菌株间的Shannon指数为0.5100。可见,河南省小麦白粉菌具有丰富的遗传多样性,研究所用的2种方法均可用于遗传多样性分析,其中AFLP分析小麦白粉菌群体表现出更为丰富的遗传多样性。     

Genotypic diversity in barley powdery mildew populations in northern France

Diversity in populations of Erysiphe graminis ( Blumeria graminis ) f.sp. hordei was studied with virulence and molecular markers. Isolates were sampled in two locations in northern France from a winter barley cultivar (Plaisant) and a spring cultivar (Caruso). Only a few pathotypes (determined by virulence markers) were common. The rest of the population was diverse. Diversity within common pathotypes, estimated by five RAPD and two SCAR markers, was generally high, except for one pathotype, which was frequent on Plaisant. This pathotype carried only one virulence, Va22, out of the 11 virulences tested. It appeared as a clonal lineage, which had occurred previously, at least in 1992, in northern France, demonstrating survival of asexual lineages in populations that often reproduce sexually.     

Genetic diversity of the pea root pathogen Aphanomyces euteiches in Europe

The oomycete pathogen Aphanomyces euteiches causes root rot in various legume species. In this study we focused on A. euteiches causing root rot in pea (Pisum sativum), thereby being responsible for severe yield losses in pea production. We aimed to understand the genetic diversity of A. euteiches in Europe, covering a north-to-south gradient spanning from Sweden, Norway and Finland to the UK, France and Italy. A collection of 85 European A. euteiches strains was obtained, all isolated from infected pea roots from commercial vining pea cultivation fields. The strains were genotyped using 22 simple-sequence repeat markers. Multilocus genotypes were compiled and the genetic diversity between individual strains and population structure between countries was analysed. The population comprising strains from Italy was genetically different and did not share ancestry with any other population. Also, strains originating from Finland and the eastern parts of Sweden were found to be significantly different from the other populations, while strains from the rest of Europe were more closely related. A subset of 10 A. euteiches strains from four countries was further phenotyped on two susceptible pea genotypes, as well as on one genotype with partial resistance towards A. euteiches. All strains were pathogenic on all pea genotypes, but with varying levels of disease severity. No correlation between the genetic relatedness of strains and virulence levels was found. In summary, our study identified three genetically distinct groups of A. euteiches in Europe along a north-to-south gradient, indicating local pathogen differentiation.     

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。     

2009—2010年河南省小麦白粉菌群体毒性及其遗传多样性分析

利用40个鉴别寄主对河南省5个小麦产区的44个单孢子堆纯化分离的小麦白粉病菌进行毒性鉴定分析,并采用多基因家系法进行了遗传多样性分析。结果显示,供试群体对Pm8、Pm1、Pm3a、Pm3b、Pm3c、Pm3f、Pm3e、Pm5、Pm6、Pm7、Pm17、Pm19、Pm34等抗病基因的毒性频率分别达到90%以上,已不能再作为抗源利用;对Pm12、Pm16、Pm21、Pm18、Pm2、Pm2＋6、Pm2＋MLD、Pm5（Mli）、Pm23、Pm30、Pm4a等基因的毒性频率低于15%,仍可以在抗病育种中加以利用;河南省的小麦白粉病菌存在地理之间的差异,同时病菌在地区之间有自西南向东北/东方向的传播。表明省内群体中存在着丰富的遗传多样性。     

甘肃省大麦条纹病菌遗传多样性及致病力差异

为了解甘肃省大麦条纹病病原菌Pyrenophora graminea的遗传多样性及致病力差异,运用RAPD分子标记技术对大麦条纹病菌不同菌株进行遗传多样性分析,并采用三明治法进行菌株致病力差异研究。结果表明:17个RAPD标记从45个菌株中扩增出126条带,平均每个标记7.41条带,遗传相似系数范围为0.468 3～0.984 1,平均值为0.830 8,当遗传相似系数为0.723 6时,可将供试菌株划分为4个类群,分别包含41、2、1和1个菌株;致病力测定结果显示菌株QWC较菌株QQ致病力强,两菌株除在品种‘甘啤2号’和‘GP-3’上无致病力外,在其他供试品种上致病力均存在差异。表明大麦条纹病菌不同菌株间存在遗传差异,且菌株QWC和菌株QQ存在致病力差异。     

Nei's gene diversity and the index of average differences are identical measures of diversity within populations

It has been established that Nei's measure of the average genetic diversity per locus, H _S , and the measure of average differences between isolates with respect to simple mismatch dissimilarity, are identical measures of diversity within populations. The Müller index of diversity can be considered as the correction of Nei's measure for small samples.     

The effect of fungicide dose on the composition of laboratory populations of barley powdery mildew

The effect of the use of different doses of the fungicide fenpropimorph on populations of barley powdery mildew Blumeria ( Erysiphe ) graminis f. sp. hordei was investigated in a laboratory selection experiment. A sample from the Danish aerial population of powdery mildew was split into populations, and these were kept separately for 31 generations on susceptible barley seedlings treated with fungicide at two concentrations, as well as on a control. Samples from these populations were tested for their resistance to fenpropimorph and their virulence spectra. There was a large amount of environmental variation in the ED₅₀ values used to measure fungicide resistance. In both treated populations, the average level of fungicide resistance increased, this increase being faster and greater in the population treated with the high dose. The diversity of pathotypes of the treated populations decreased, with the decline being more rapid in the population treated with the high dose, where one pathotype dominated the population after 31 generations. This pathotype was apparently not the fittest in the population treated with the low dose. This implies that knowledge of ED₅₀ is not sufficient to predict pathotype evolution under different fungicide treatments. The dominant pathotype in the high-dose treatment may not have been clonal, as there was evidence of two levels of fungicide resistance. The large environmental variation observed in estimated ED₅₀ values for resistance towards fenpropimorph may help to explain why this resistance has evolved at a slower rate than resistance towards other fungicides.     

Virulence spectrum in populations of the barley powdery mildew pathogen, Erysiphe graminis f. sp. hordei in Tunisia and Morocco in 1992

The virulence of 57 single colonies of Erysiphe graminis f. sp. hordei from Tunisia and Morocco was investigated using the Pallas near-isogenic differential set. Virulence patterns in general were similar in both countries. Va8, Va10+Du2, V41/145, VLa and VRu2 appear to be common in the region. The resistance alleles Mla7 and Mla9, in combination with other resistances, were highly effective against the isolates tested. No virulence against mlo and Mla9 + k was detected in Tunisia. No virulence on Mla7 and Mla9 was detected in Morocco. The frequency of virulence against several resistances was significantly higher in Morocco than in Tunisia. On the other hand, virulence against other resistances was higher in Tunisia than in Morocco. Three isolates from Morocco infected mlo to an extent greater than previously described. All pathotypes were unique. An attempt is made to interpret the results by comparison with pathotype evolution in Europe.     

南宁市桑白粉病病原菌种类鉴定

桑白粉病是桑树的重要病害,分为桑里白粉病和桑表白粉病。通过形态特征观察和ITS、D1/D2序列分析,明确桑里白粉病的病原菌为桑生球针壳Phyllactinia moricola;桑表白粉病的病原菌为桑白粉菌Erysiphe mori。桑钩丝壳Uncinula mori是桑白粉菌的异名。     

Population genetic structure of four regional populations of the barley pathogen Pyrenophora teres f. maculata in Iran is characterized by high genetic diversity and sexual recombination

The leaf spot form of the barley disease net blotch, caused by the fungus Pyrenophora teres f. maculata (PTM), is an increasingly important foliar disease of barley. Studies of population genetic structure and reproductive mode are necessary to make predictions of the evolutionary potential of the pathogen. Sources of resistance to PTM have been found in Iranian landraces, which may have the potential to improve plant breeding efforts. However, little is known about the population genetic structure of this fungus in Iran. In this study, we analysed the frequency of the mating type genes to assess the potential for sexual mating of PTM collected from four provinces—Khuzestan, Hamadan, Golestan, and East Azerbaijan—and we investigated the population genetic structure using seven simple sequence repeat markers. High genotype diversity, linkage equilibrium, and equal ratios of mating types frequencies in the PTM populations at Khuzestan and Hamadan support the occurrence of sexual reproduction in these populations, while in Golestan and East Azerbaijan populations, significant gametic disequilibrium and relatively low genotype diversity suggest a higher incidence of clonality or different demographic histories. Unequal mating type frequencies in Golestan confirm a predominance of asexual reproduction. Finally, we found significant evidence for strong population structure with most of the genetic variation represented within regional populations (89%). Overall, our study provides evidence for high genetic variation in Iranian PTM populations, which may be the basis for rapid adaptive evolution in this pathosystem. This highlights the need for integrated efforts to control the disease.     

哈尔滨地区小天蓝绣球白粉病病原菌鉴定

小天蓝绣球Phlox drummondii Hook., 又名雁来红?金山海棠?福禄考, 是一种极具观赏性的一年生草本植物, 在我国北方被用来城市绿化?在小天蓝绣球的生长过程中, 白粉病已成为其所患病害中发生最为严重的一种, 给小天蓝绣球产业造成了巨大的损失, 也使其丧失了美化城市的功能?本研究以东北农业大学园艺实验站的小天蓝绣球病叶为材料, 对其病原菌进行形态学和分子生物学鉴定, 明确其分类地位, 为小天蓝绣球白粉病的科学防治提供依据?结果表明:小天蓝绣球白粉病菌主要侵染叶片, 叶片正面布满白色粉层, 后期叶片上产生黑褐色的小点, 即闭囊壳?闭囊壳直径80~160 μm, 附属丝菌丝状, 长度30~210 μm, 子囊多个, 一个子囊内含有子囊孢子2~8个, 分生孢子为粉孢子?对病原菌的ITS序列进行PCR扩增, 得到561 bp的序列(GenBank登录号:MN209867), 在GenBank上进行BLAST比对, 确定此病菌为Golovinomyces magnicellulatus?     

黑龙江省番茄白粉病病原鉴定

通过收集东北农业大学日光温室中的番茄白粉菌,并对其进行形态和分子鉴定及分析,明确了该白粉病的病原种类及其分类地位。根据分生孢子的显微形态观察初步确定为新番茄粉孢菌（Oidiumneolycopersici L. Kiss）。进一步根据真菌rDNA-ITS序列检测分析后发现该病原菌与新番茄粉孢菌（O. neolycopersici）的相似性为100%,并对与其相似性较高的序列构建进化树分析发现,本试验研究的白粉菌与其他国家和地区的白粉菌相似性不高,这可能是由于哈尔滨地区白粉菌自身小种进化的原因。     

Polygenic powdery mildew disease resistance in Arabidopsis thaliana: quantitative trait analysis of the accession Warschau-1

To further the understanding of the natural genetic diversity for disease resistance to powdery mildew ( Erysiphe cichoracearum ) in Arabidopsis thaliana , quantitative trait loci analysis was undertaken on recombinant inbred lines derived from a cross between the resistant accession Warschau-1 and the susceptible Columbia-0. Powdery mildew grew less well on Warschau-1, but the resistance was not associated with a specific block in the infection sequence. Two potential powdery mildew disease-resistance loci were identified and mapped, one with a major effect and one with a minor effect on disease resistance. The two loci acted in an additive manner to confer resistance, and together they explained 65% of the variation in resistance. In addition, the major powdery mildew disease-resistance locus was genetically mapped to the bottom of chromosome III, a region containing the powdery mildew resistance loci RPW7 , RPW8 and RPW10 . Unlike resistance mediated by the RPW8 locus in the accession Moscow-1, resistance in Warschau-1 was not correlated with the hypersensitive response, highlighting the influence of genetic background or environmental factors on the expression of disease resistance. Together with the powdery mildew resistance loci described in other studies, these results suggest that A. thaliana is a useful source of natural powdery mildew disease resistance, which potentially can be utilized in fundamental studies and as a tool for applied studies.     

河南省小麦黑胚病优势病原菌链格孢菌(Alternaria spp.)遗传多样性分析

 采用RAPD技术对分离自河南省各地的43株小麦黑胚病优势病原菌(Alternaria spp.)菌株进行了遗传多样性分析。17个随机引物共扩增出151条清晰的DNA条带,所扩增出的DNA条带均为多态带,说明河南省小麦黑胚病菌存在着丰富的遗传多样性。利用NTSYS软件进行了病原菌的聚类分析,结果表明,河南省小麦黑胚病菌主要有2个种,即Alternaria alternata和A. tenuissima,种间的遗传相似系数的变化幅度为0.62~0.92。来自同一个地区的小麦黑胚病菌菌株基本上聚在了一起,表现出很近的亲缘关系,来自不同地区之间的菌株也可以交叉聚类。病原菌遗传多样性分析进一步验证了形态学的鉴定结果。     

High levels of genetic and genotypic diversity in field populations of the barley pathogen Ramularia collo-cygni

The ascomycete pathogen Ramularia collo-cygni causes Ramularia leaf spot (RLS) on barley. Although R. collo-cygni is considerd an emerging disease of barley, little is known about genetic diversity or population genetic structure of this pathogen. We applied a set of polymorphic AFLP (Amplified Fragment Length Polymorphism) markers to investigate population genetic structure in two Northern European populations of R. collo-cygni. The distribution of AFLP alleles revealed low levels of population subdivision and high levels of genetic diversity at both locations. Our analyses included 87 isolates and of these 84 showed a unique genotype pattern. The genetic structure of populations in Scotland and Denmark is highly similar and we find no evidence of population sub-division. An analysis of molecular variance was used to show that 86 % of the variance is attributable to within field genetic variance. In spite of the high levels of genetic and genotypic diversity in the R. collo-cygni populations, we find significant evidence of linkage disequilibrium among the AFLP alleles using a multilocus analysis. We propose that the high levels of genotypic diversity and the lack of population differentiation result from considerable levels of gene flow between populations most likely mediated by seed borne dispersal of inoculum.     

Pathogenic and molecular diversity in highly virulent populations of the parasitic weed Orobanche cumana (sunflower broomrape) from Europe

The parasitic weed Orobanche cumana (sunflower broomrape) constrains sunflower production in eastern and southern Europe and in the Middle East. Although genetic resistance is the most effective control method, new parasite races evolve overcoming sunflower resistance. In this work, highly virulent populations of O. cumana were analysed for pathogenicity and genetic diversity. The virulence of 11 populations from Hungary, Romania, Spain and Turkey was assessed and compared after infection of sunflower inbred lines to differentiate races of the parasite under glasshouse conditions. Molecular diversity among and within 27 parasite populations was studied by RAPD‐PCR, UPGMA and amova analyses. Highly virulent race F was identified in Hungary, Spain and Turkey. The most virulent race (G) was also found in Turkey. The molecular analysis among highly virulent populations of O. cumana identified four molecular clusters, respectively, grouping populations from Central Spain, Hungary, South Spain and Turkey. The genetic homogeneity within parasite populations was confirmed, since no molecular divergences were found within them. This work constitutes the first geographical study of O. cumana together with pathogenicity and molecular traits inherent to each geographical group, and provides useful information for possible phylogenetic analyses of O. cumana. In addition, molecular markers associated with geographical origin could be developed and used as diagnostic tools to track new broomrape introductions into areas free of virulent races where they might represent a threat to sunflower production.     

西南地区稻瘟病菌群体遗传多样性分析

为明确西南地区稻瘟病菌Magnaporthe grisea(Hebert)Barr群体遗传结构及其多样性水平,选用13对SSR引物对来自18个县(市)的221个稻瘟病菌单孢菌株进行PCR扩增,利用最长距离法和生物学软件进行聚类分析和群体遗传多样性分析。结果显示,13对SSR引物均能扩增出一条大小相同且清晰的条带,多态位点百分率高达100%。221个菌株在0.16相异水平上可划分为13个遗传宗谱,宗谱SCL01含205个菌株,占总菌株数的92.76%,为优势宗谱;宗谱SCL02~SCL013为劣势宗谱,差异极大。在群体水平上,菌源丰富的8个区域稻瘟病菌群体的Nei’s基因多样性指数为0.2133,Shannon信息指数为0.3588,具有丰富的遗传多样性,且群体间差异较大;这8个种群基于UPGMA法大都聚为一类,种群遗传谱系与地理区域分布呈一定相关性,群体遗传多样性均值为0.2518,存在一定的遗传分化,且群体内多样性大于群体间,总遗传变异的59.37%存在于群体内。总体上,西南地区稻瘟病菌群体结构既有明显的优势宗谱,又存在许多复杂多变的特异性小宗谱,具有丰富的遗传多样性,与地理分布关系较为密切。     

Genetic variation in populations of the cacao wilt pathogen, Ceratocystis cacaofunesta

Ceratocystis cacaofunesta (=  Ceratocystis fimbriata ) causes a lethal wilt disease of cacao ( Theobroma cacao ) in Latin America. Polymorphic microsatellite markers, (CAT)₅ nuclear DNA fingerprints and Hae III mitochondrial DNA fingerprints were used to compare genetic diversity among isolates of C. cacaofunesta collected from populations in western Ecuador, Costa Rica, Colombia, and Rondônia and Bahia in Brazil. Microsatellite markers and nuclear DNA fingerprints separated Ecuadorian isolates from isolates of the other four populations, and these two major groups correspond to genetic lineages already identified from ITS-rDNA sequences and intersterility groupings. Mitochondrial DNA fingerprints also demonstrated substantial diversity and split the Ecuadorian isolates into two groups. All marker types showed limited variation in the Colombian, Costa Rican and Bahian populations, as might be expected for introduced populations that have gone through recent genetic bottlenecks. In contrast, the Rondonian and western Ecuadorian populations showed gene diversity values similar to natural populations of other Ceratocystis species. The Rondonian population was the only sampled population in the native range of T. cacao (the Upper Amazon), and the putatively introduced populations were more closely related to the Rondonian population than to the western Ecuadorian population. The Ecuadorian population is in an area with other native Theobroma species, which may serve as natural hosts.     

我国烟草赤星病菌遗传多样性的ISSR分析

为明确我国烟草赤星病的2种主要致病菌链格孢菌Alternaria alternata和长柄链格孢菌A.longipes的地理差异与遗传结构,采用ISSR标记对分离自9个省市的135株烟草赤星病菌进行遗传多样性分析。结果显示,通过正交优化试验建立的烟草赤星病菌ISSR-PCR最佳反应体系稳定性较好,筛选出17条多态性高且稳定的引物,共扩增出192条谱带,其中有177条具有多态性,多态率为92.19%。UPGMA聚类分析结果显示,链格孢菌和长柄链格孢菌的遗传相似性系数分别在0.67~1.00和0.66~1.00之间,遗传相似性系数为0.83时可使链格孢菌和长柄链格孢菌分别划分为5个和6个亚群,其中前者不同地理种群间表现出地理相关性,后者不同菌株随机分组。烟草赤星病菌种群的基因多态性和遗传多样性丰富,链格孢菌和长柄链格孢菌的群体间遗传分化系数分别为0.36和0.37,均存在遗传分化;群居每代迁移数分别为0.89和0.85,不同地理种群间存在基因交流;2种烟草赤星病菌的遗传分化结构表现出相似性。表明我国烟草赤星病菌中的链格孢菌和长柄链格孢菌均存在丰富的遗传多样性,且二者进化方向相似,ISSR标记能较好地揭示烟草赤星病菌种群间的亲缘关系和遗传差异性,可用于其遗传多样性分析。