陇南地区不同海拔区域内小麦条锈菌群体遗传多样性的分析

小麦条锈病是世界范围内小麦上最重要的流行性病害之一,可造成严重的产量损失。陇南地区是我国小麦条锈菌主要越夏易变区和新小种发源地,了解该地区不同海拔高度区域内条锈菌遗传多样性有重要意义。本研究采用TP-M13-SSR荧光标记技术对11个种群330个小麦条锈菌分离株基因组DNA进行了SSR标记分析。不同海拔区域的条锈菌遗传多样性有明显的差异,高山区的遗传多样性比较丰富,半山区次之,川道区相对比较低。不同生态区域内,小麦条锈菌群体遗传分化程度不同,高山区和半山区遗传分化程度大,基因流小,川道区群体遗传分化程度比较小,基因流大。来自不同海拔区域的菌系具有相同的基因型,这一结果从分子水平证明了在陇南地区小麦条锈菌在山区与川地之间存在广泛的菌源交流,可就地完成周年循环。     

麦无网长管蚜不同地理种群遗传多样性的ISSR标记研究

为阐明麦无网长管蚜Metopolophium dirhodumus不同地理种群的遗传多样性,利用ISSR分子标记对6个地区（河北保定、定州、石家庄、邢台、邯郸,山东聊城）的麦无网长管蚜种群进行了遗传多样性研究。23条ISSR引物扩增出了297条清晰条带,其中172条（57.91%）具有多态性。遗传多样性分析结果表明：麦无网长管蚜群体间的遗传多样性（57.91%）高于群体内的遗传多样性（26.24%）,其中河北保定种群和河北邯郸种群的种群内遗传变异最小（24.92%）,河北石家庄种群的种群变异最大（41.75%）,河北和山东地区的种群（除河北石家庄）之间没有显著遗传差异。根据种群变异来源分析,有26.44%遗传变异来源于种群间,73.56%变异来自于种群内（Gst=0.2644）,不同地理种群间并没有出现分化现象（Nm=1.3910）。利用不加权算术平均法（UPGMA）对6个种群进行统计分析,构建进化树,发现6个种群分为了两大类,而其地理距离与遗传距离并不存在相关性,地理空间并未对其种群间的基因交流产生影响。麦无网长管蚜有较高的遗传多样性,而各个种群间并没有明显遗传分化的趋势。本研究有关麦无网长管蚜不同地理种群基因流和遗传变异将为控制麦蚜危害和田间综合防治提供必要的数据支持。     

甘肃天水地区小麦条锈菌自然群体DNA指纹分析

对1997—1999连续三年采自甘肃省天水地区16个地点的244个小麦条锈菌分离系进行了PSR331S3/Bg1Ⅱ指纹分析,共鉴定出表现型185个,遗传多样性Shannon指数M=4.467, 加权修正值M*=0.9106,显现出高度的遗传多样性水平;其中三个主要采样点凤凰、甘谷和麦积山病菌群体M值分别为3.5720、3.0268和3.4186,加权修正值M*分别为0.9330、0.8513和0.8610。三个群体之间的遗传分化系数GST=0.03167,显示出较低的遗传分化水平。本文以多拷贝的RFLP标记为手段,揭示了小麦条锈菌作为一种活体寄生的远程气传病原真菌,其越夏栖息地关键区(天水地区)自然群体存在丰富的遗传多样性,不同海拔生境对病菌群体结构有显著的影响。但是,地区内亚群体间分化水平很低,表明区域内菌源交流十分频繁。     

小菜蛾不同地理种群遗传多样性的ISSR标记研究

为阐明小菜蛾Plutella xylostella不同地理种群的遗传多样性, 应用ISSR技术对我国小菜蛾8个地理种群的遗传多样性进行了研究分析。15条引物扩增出395条ISSR条带, 其中多态性条带占89.11%, 全部个体显示了各自独特的ISSR图谱。ISSR标记的遗传多样性分析结果表明： 小菜蛾无论在物种水平上（P=89.11%, H=0.2706, I=0.4286）, 还是在种群水平上（P=88.80%, H=0.2759, I=0.4349）都表现出较高的遗传多样性。其中, 北京南口种群内遗传变异最大, 海南海口和甘肃兰州种群内遗传变异最小, 南方地区（云南、 湖北）小菜蛾种群遗传多样性明显高于北方地区（北京、 天津、 山东、 黑龙江、 甘肃）种群。据种群变异来源分析, 有5.66%的遗传变异来自种群间, 94.34%的变异来源于种群内（N_m=8.3399）, 不同地理种群间没有明显的遗传分化。本文有关小菜蛾不同地理种群基因流动和遗传变异的研究为小菜蛾抗药性的控制及田间种群的综合防治提供了有价值的分子生物学依据。     

附子野生资源群体遗传多样性的RAPD分析

应用RAPD标记分析了分布在附子主栽区四川、重庆、陕西及湖北16个野生乌头种群的遗传变异。24个引物共检测到643个RAPD位点,多态位点602个,总的多态位点百分率达93．5％。Shannon多样性和Nei遗传分化结果一致显示重庆酉阳种群和重庆城口种群遗传多样性最高,四川盐源种群和陕西勉县种群的遗传多样性最低。Shannon指数测出的种群内的遗传变异（57．6％）略占优势,群体间的遗传分化达到42．4％;Nei基因分化系数（GST）达40．0％;分子方差分析（AMOVA）发现群体间遗传变异仅为25．37％;种群每代迁移数Nm为0．756。Nei相似性系数的UPGMA分析结果显示该地区的野生乌头分布上有一定的地域性,特别是同为附子道地产地江油供种的北川、安县和青川种群间遗传关系密切,说明种质资源在道地药材形成中具有重要作用。研究结果表明,附子主要栽培地区的乌头野生种群之间存在较大的遗传分化,遗传多样性较高,遗传种质资源较丰富,存在一定的特异性资源,为进一步开发利用乌头（川乌、附子）提供了丰富的种质资源。     

大别山山核桃种群遗传多样性研究

为了更有效地保护和合理开发大别山山核桃(Carya dabieshanensis)资源,该文利用RAPD分子标记技术,对3个天然大别山山核桃种群的90个单株的遗传多样性、种群内和种群间的遗传变异进行了研究,结果表明:20对10 bp随机引物共检测到238条谱带,其中多态带为162条,占68.1%。遗传多样性分析结果显示: Shannon多样性指数为0.476 1,58.18%的变异分布于群体内,而种群间变异占了41.82%;Nei指数群体总基因多样度为0.314 5,群体内平均基因多样度(H_S)为0.186 5,群体间的基因多样度(H_ST)为0.128 0,群体Nei基因分化系数(G_ST)为0.406 7,说明40.67%的变异存在于种群间,群体内的变异占了总变异的59.33%,与Shannon多样性指数相比基本一致,均表明种群内有较丰富的遗传变异,这为优良品种选育提供广阔前景;种群间的基因流(N_m)为0.730 6,证明种群间遗传交换较小,这与环境适应性和高山阻隔有一定的关系。     

内蒙古地区亚洲小车蝗不同地理种群的RAPD分析

采用随机扩增多态性DNA(RAPD)技术对内蒙古地区亚洲小车蝗Oedaleus asiaticus(B.-bienko)9个不同地理种群90个个体进行扩增,8条随机引物扩增共产生了78条带,多态性片段为62条。对Nei′s基因多样性指数和遗传距离进行分析,结果表明:种群间的遗传分化系数为0.2343,即23.43%的遗传变异存在于种群间,种群内的遗传分化系数为0.7657,即76.57%的遗传多样性存在于种群内,群体内遗传多样性大于群体间遗传多样性。用NJ法对这3个种群的Nei′s遗传距离作聚类分析,结果表明亚洲小车蝗不同种群的遗传分化程度与地理距离具有正相关关系。     

濒危灌木长叶红砂遗传多样性的RAPD分析

应用RAPD分子标记对濒危灌木长叶红砂(Reaumuria trigyna)种群遗传多样性进行了分析.应用18条随机引物对长叶红砂5个种群的95个个体进行扩增,检测到118个位点,其中多态位点105个.结果表明:长叶红砂种群的多态位点比率(P)为88.98%,显示出长叶红砂种群存在较高的遗传多样性.Shannon多样性指数(0.4966)、Nei基因多样性指数(0.3303)和基因分化系数(Gst=0.1425)的分析结果显示,长叶红砂种群遗传变异大多存在于种群内,种群间的遗传分化占14.25%.聚类分析表明,长叶红砂种群遗传距离与地理距离之间无直接相关关系.遗传多样性水平与物种特性和所处不同群落有关,濒危植物并不一定表现为遗传变异水平的降低.     

基于线粒体COI序列变异的高原鼢鼠种群遗传分化研究

基于线粒体COI基因序列对高原鼢鼠7个地理种群的遗传多样性、遗传分化和基因流进行分析,158条COI基因序列中发现了570个变异位点,界定了54个COI单倍型。总体单倍型多样性指数Hd为0.911,种群单倍型多样性在0.278—0.964之间,高原鼢鼠的种群遗传多样性较高。群体总的遗传分化系数Fst为0.611,群体间的基因流Nm在-0.020—3.700之间,部分地理种群之间的遗传分化程度高,基因流弱。AMOVA分子变异分析显示,高原鼢鼠的遗传变异主要来自种群之间(77.56%),种群内部的遗传变异较低(22.44%)。中性检验(P0.01)与错配分析显示高原鼢鼠种群经历了群体扩张事件。IBD未检测到地理距离与种群间遗传距离的显著性相关关系(R2=0.124,P=0.118)。综合分析认为,高原鼢鼠不同地理种群间遗传分化的原因除地理隔离外,还与迁移扩散方式、进化过程和其他环境因素有关。     

钱塘江日本沼虾野生群体遗传变异的SSR分析

利用微卫星标记分析了钱塘江干流水域闻堰、富阳、场口、桐庐等7个野生日本沼虾群体的遗传多样性和遗传结构。结果表明:10个微卫星位点呈现高度多态性;闻堰、富阳、场口、新安江等中下游野生群体的遗传多样性水平有高于歙县和休宁两个上游群体遗传多样性水平的趋势。符号检验和Wilcoxon符号秩次检验的结果表明,钱塘江日本沼虾群体近期没有发生瓶颈效应,群体数量也没有下降。FST的范围介于0.0201～0.1069。分子方差分析结果显示,大部分的遗传变异(93.48%)存在于个体间,少部分遗传变异(6.52%)存在于群体之间。群体间FST及AMOVA分析表明,群体处于中等遗传分化水平;基于DA遗传距离构建的NJ聚类树显示,地理位置相邻的群体聚在一起。STRUCTURE分析413份参试的日本沼虾样本被分为2个理论种群,即上游歙县和休宁群体为一个理想种群,其余中下游的5个群体为另一个理想种群。日本沼虾的遗传多样性和遗传结构与所生存的地理位置具有相关性。     

中国小麦条锈菌转主寄主小檗的鉴定

用萌发的小麦条锈菌冬孢子接种采自陕西省境内的陕西小檗、少齿小檗和长穗小檗,3种小檗均产生了性孢子器和锈孢子器。用人工接种小麦条锈菌冬孢子在陕西小檗上产生的锈孢子器接种小麦铭贤169产生了典型的条锈菌夏孢子堆症状。特异性PCR和DNA序列分析表明,人工接种产生于小檗上的锈孢子、接种锈孢子于小麦上产生的夏孢子堆与小麦条锈菌DNA的ITS区序列完全一致。更为重要的是,用采自田间受锈菌侵染的小檗叶片产生的锈孢子接种小麦铭贤169,经培养在小麦铭贤169叶片上产生了典型的条锈病症状。从而证实,在自然条件下,在中国,小檗不仅可作为小麦条锈菌的转主寄主,而且小麦条锈菌可在小檗上完成其有性繁殖过程。这一发现对进一步揭示我国小麦条锈菌高度的群体遗传多样性与毒性变异机理、完善小麦条锈病的防治策略具有十分重要的理论和实际意义。     

小麦条锈菌条中31号生理小种SCAR检测标记的建立

建立小麦条锈菌Pucciniastriiformisf.sp.tritici生理小种的快速分子检测技术对我国小麦条锈病的监测和防治策略的制定具有重要价值,本文首次报道了利用SCAR—PCR技术进行条锈菌生理小种分子检测的方法。通过对我国目前主要优势小种条中31号RAPD片段的规模筛选,在对特异片段回收、克隆、测序的基础上,设计特异PCR引物,成功获得了条中31号生理小种专化的SCAR检测标记。     

小麦条锈菌分子生态学研究进展

小麦条锈病是危害最严重的小麦流行性病害之一,小麦条锈菌的生态学研究对制定合理的防治策略和抗锈育种具有重要意义.近十几年来,DNA分子标记技术被应用于小麦条锈菌的群体遗传学研究,推动了小麦条锈菌分子生态学研究的快速发展,为揭示小麦条锈菌的群体生态特性开辟了一个新的途径.本文系统介绍小麦条锈菌分子生态学研究的主要进展,并就我国当前研究的局限性和发展趋势进行了分析.     

Comparative virulence phenotypes and molecular genotypes of Puccinia striiformis f. sp. tritici, the wheat stripe rust pathogen in China and the United States

Stripe rust (yellow rust) of wheat, caused by Puccinia striiformis f. sp. tritici, is one of the most important diseases in both China and the United States. The Chinese and US populations of the stripe rust fungus were compared for their virulence phenotypes on wheat cultivars used to differentiate races of the pathogen in China and the US and molecular genotypes using simple sequence repeat (SSR) markers. From 86 Chinese isolates, 54 races were identified based on reactions on the 17 Chinese differentials and 52 races were identified based on the 20 US differentials. The selected 51 US isolates, representing 50 races based on the US differentials, were identified as 41 races using the Chinese differentials. A total of 132 virulence phenotypes were identified from the 137 isolates based on reactions on both Chinese and US differentials. None of the isolates from the two countries had identical virulence phenotypes on both sets of differentials. From the 137 isolates, SSR markers identified 102 genotypes, of which 71 from China and 31 from the US. The virulence data clustered the 137 isolates into 20 virulence groups (VGs) and the marker data clustered the isolates into seven molecular groups (MGs). Virulence and SSR data had a low (r = 0.34), but significant (P = 0.01) correlation. Principal component analyses using either the virulence data or the SSR data separated the isolates into three groups: group a consisting of only Chinese isolates, group b consisting of both Chinese and US isolates and group c consisting of mostly US isolates. A neighbour-joining tree generated using the molecular data suggested that the P. striiformis f. sp. tritici populations of China and the US in general evolved independently.     

Detection of Puccinia striiformis in Latently Infected Wheat Leaves by Nested Polymerase Chain Reaction

Stripe rust, caused by Puccinia striiformis f. sp. tritici ( Pst ), is one of the most devastating wheat diseases worldwide, especially in temperate regions with cool moist weather conditions. The identification of the pathogen in infected plants based on morphological or physiological criteria before sporulation is labour-intensive and time-consuming. To accelerate and simplify the process of detection, a nested Polymerase Chain Reaction (PCR) assay was developed for specific and sensitive detection of Pst . Specific primers Psta-Psts were designed according to a genome-specific sequence of Pst . In nested PCR, with a 10-fold dilution series of template DNA, the detection limit was 2 pg DNA in the first PCR with the primers Psta-Psts. The second round PCR was then performed using amplified products from the first PCR as the template and Nesta-Nests as the primers. An amplification signal was detectable even when only 2 fg of P. striiformis f. sp. tritici DNA was used as the template in nested PCR. With nested PCR, the sensitivity of detection was enhanced 1000 fold. Using extracts from stripe rust-infected wheat leaves, the fungus could be determined in the leaves before symptom appearance. The assay provides a rapid and sensitive method for detection of P. striiformis f. sp. tritici in latently infected leaves of overwintering wheat plants.     

Cloning and characterization of the actin gene from Puccinia striiformis f. sp. tritici

The fungus Puccinia striiformis f. sp. tritici, the causal agent of wheat stripe rust, is an obligate biotrophic basidiomycete. Urediniospores are the most common spore type involved in the epidemiology of this disease. Tip growth of germ tubes of germinated urediniospores is a key step during infection of wheat, but few studies have investigated it so far. Recent research has found that actin is closely associated with hyphal tip growth. In this study, we have cloned and obtained the full-length actin cDNA from P. striiformis f. sp. tritici and characterized its expression. Furthermore, actin filament (F-actin) patterns were visualized microscopically during germ tube formation. The most conspicuous actin-containing structures were actin patches. They were mainly concentrated near the hyphal tip and scattered throughout the cortex. By using cytochalasin B, we observed that depolymerization of F-actin greatly reduced the germination rate of urediniospores and disrupted the transport of vesicles to the germ tube tip, indicating that F-actin played a key role in the tip growth of P. striiformis f. sp. tritici. This work helps us to understand the tip growth mechanism of P. striiformis f. sp. tritici, and may provide a theoretical framework for designing novel pesticides.     

Nonhost resistance of rice to rust pathogens

Rice is atypical in that it is an agricultural cereal that is immune to fungal rust diseases. This report demonstrates that several cereal rust species (Puccinia graminis f. sp tritici, P. triticina, P. striiformis, and P. hordei) can infect rice and produce all the infection structures necessary for plant colonization, including specialized feeding cells (haustoria). Some rust infection sites are remarkably large and many plant cells are colonized, suggesting that nutrient uptake occurs to support this growth. Rice responds with an active, nonhost resistance (NHR) response that prevents fungal sporulation and that involves callose deposition, production of reactive oxygen species, and, occasionally, cell death. Genetic variation for the efficacy of NHR to wheat stem rust and wheat leaf rust was observed. Unlike cereal rusts, the rust pathogen (Melampsora lini) of the dicotyledenous plant flax (Linum usitatissimum) rarely successfully infects rice due to an apparent inability to recognize host-derived signals. Morphologically abnormal infection structures are produced and appressorial-like structures often don't coincide with stomata. These data suggest that basic compatibility is an important determinate of nonhost infection outcomes of rust diseases on cereals, with cereal rusts being more capable of infecting a cereal nonhost species compared with rust species that are adapted for dicot hosts.     

Molecular markers for leaf rust resistance genes in wheat

Over 100 genes of resistance to rust fungi: Puccinia recondita f. sp. tritici, (47 Lr - leaf rust genes), P. striiformis (18 Yr - yellow rust genes) and P. graminis f. sp. tritici (41 Sr - stripe rust genes) have been identified in wheat (Triticum aestivum L.) and its wild relatives according to recent papers. Sixteen Lr resistance genes have been mapped using restriction fragments length polymorphism (RFLP) markers on wheat chromosomes. More than ten Lr genes can be identified in breeding materials by sequence tagged site (STS) specific markers. Gene Lrk 10, closely linked to gene Lr 10, has been cloned and its function recognized. Available markers are presented in this review. The STS, cleaved amplified polymorphic sequence (CAPS) and sequence characterized amplified regions (SCAR) markers found in the literature should be verified using Triticum spp. with different genetic background. Simple sequence repeats (SSR) markers for Lr resistance genes are now also available.     

Rates of evolution of avirulence phenotypes and DNA markers in a northwest European population of Puccinia striiformis f. sp. tritici

The effects of evolutionary processes in fungal pathogen populations may occur more rapidly and display larger effects in agricultural systems than in wild ecosystems because of human involvement by plant breeding and crop management. In this study, we analysed the rate of evolution in three lineages of a northwest European population of a biotrophic and asexual reproduced fungal pathogen, Puccinia striiformis f. sp. tritici, causing yellow rust on wheat. Pathogen samples were collected between 1975 and 2002 in the UK and Denmark, and assayed for 14 individual avirulence/virulence alleles and up to 234 amplified fragment length polymorphism (AFLP) primer pairs producing approximately 17,000 AFLP fragments. The large number of fragments and a targeted sampling of isolates allowed a reconstruction of phylogenies in great detail, i.e. no homoplasy and a representation of sequential, evolutionary steps by pathogen samples. A recent, phenotypic loss of avirulence was observed at least once for loci corresponding to P. striiformis f. sp. tritici resistance Yr2, Yr3, Yr4, Yr7, Yr9, and Yr15, whereas Avr6 and Avr17 were lost independently in all three lineages, corresponding to 16 events of loss of avirulence (emergence of virulence). The opposite process, restoration of avirulence, was observed for Yr9 and Yr32. An interpretation of phenotypic changes within lineages as independent mutation events resulted in mutation frequencies from 1.4x10(-6) to 4.1x10(-6) per AFLP fragment (locus) per generation, whereas the effective rate by which a mutation from avirulence to virulence was established in the pathogen population, when subject to selection by host resistance genes, was approximately three orders of magnitude faster.