共查询到18条相似文献，搜索用时 939 毫秒
1.
小麦条锈菌水源11类群的RAPD分析及SCAR标记的建立 总被引：2，自引：0，他引：2
鉴于水源11类群近年来一直处于优势地位,为简化其检测手段,本研究利用RAPD技术对该类群的8个主要致病类型进行了多态性分析,以寻找其中主要流行类型的特异性分子标记。结果如下:共筛选出10个碱基随机引物190条,其中94条可得到稳定清晰的扩增图谱,用该94条引物进行RAPD分析,发现各致病类型间遗传变异丰富;以引物S1410扩增得到了水源11-4的特异性DNA条带;以引物S1412和S1304扩增得到了水源11-14的特异性DNA条带;对引物S1304扩增得到的特异性DNA条带回收、克隆和测序,设计了1对19bp/18bp的引物,并成功地将其转化为对水源11-14特异的SCAR标记。以上结果表明,通过规模筛选来寻找小麦条锈菌生理小种的特异性DNA片段,并将其转化为稳定的SCAR标记,有可能建立起中国小麦条锈菌流行生理小种的快速分子鉴定体系。 相似文献
2.
小麦条锈菌新菌系V26的SCAR检测标记 总被引：2，自引：0，他引：2
建立小麦条锈菌生理小种的快速分子检测技术体系对小麦条锈菌的监测和防治策略的制定具有重要价值。条锈菌V26是近年来出现的,对我国目前小麦抗病育种中普遍应用的抗条锈病基因Yr26具有毒性的新菌系。该菌系的出现,对我国当前小麦生产、抗病育种都造成了严重威胁。本研究选用189条随机引物对CYR29、CYR31、CYR32、CYR33、T4、Su11-4和V26等7个条锈菌生理小种(菌系)进行了扩增,筛选V26的特异性RAPD片段,并对其进行克隆和测序。根据测序结果,设计并合成SCAR特异性引物, 将V26的RAPD标记转化为稳定的SCAR标记。使得对该菌系的快速检测成为可能,同时也将会为条锈菌新小种的监测提供更为准确的科学依据。 相似文献
3.
小麦叶锈病由Puccinia triticina引起,是我国小麦生产上的重要病害。本研究旨在建立小麦叶锈菌快速、准确的PCR诊断检测技术体系,用于病害精准测报和综合防控。以真菌β-微管蛋白基因的保守序列为引物,进行小麦锈菌gDNA的PCR比较分析,发现小麦叶锈菌具有长度为268bp的特异性DNA片段;序列分析后设计了2对专化性引物,成功获得检测灵敏度为5.00pg/μL模板DNA浓度水平的小麦叶锈菌种的特异性SCAR标记。对55个来自我国不同麦区的小麦叶锈菌标样以及其它麦类病原真菌的检测表明,该叶锈菌标记的检测可靠性达100%。人工接种条件下,叶锈菌侵染24h后,即可在小麦叶片内检测到该标记。 相似文献
4.
欧洲小麦品种Mega抗条锈病基因的遗传分析及分子标记 总被引：1，自引：0，他引：1
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。 相似文献
5.
一个新的与稻瘟病菌无毒基因AVR-Pikm紧密连锁的SCAR标记 总被引：3，自引：0，他引：3
无毒基因是病原物中决定寄主抗病性表达的功能基因,其功能的丧失导致毒性小种的产生。本研究利用随机引物扩增DNA多态性技术,从稻瘟病菌菌株S1522中新筛选到1个与无毒基因AVR-Pikm紧密连锁的DNA标记OPE121400。根据OPE121400的核苷酸序列,设计了1对含有17个核苷酸的特异性SCAR引物,并利用该引物对无毒表型亲本S1522和毒性表型亲本S159及其有性杂交后代的108个菌株进行了PCR扩增。结果表明:所有无毒表型的菌株均能特异性扩增出1条与OPE121400大小相近的DNA片段,而毒性表型的菌株除2个重组体外,均不能扩增出此片段。根据计算,这一SCAR标记与目标无毒基因AVR-Pikm之间的遗传距离为1.89 cM,与本研究小组先前报道的另一个标记OPO121000位于目标基因的同一侧,但与OPO121000相比,距目标基因近了2.86 cM。本标记的获得将有助于确定AVR-Pikm在染色体上的位置,有助于确定用于进一步筛选位于相反一侧的连锁标记的重叠群区域。 相似文献
6.
7.
以小麦优异抗源Sw92为父本,感病小麦品种铭贤169为母本,杂交获得F1、F2和BC1代群体。采用我国叶锈菌优势小种PHT对双亲及其杂交世代进行接种鉴定。结果表明,小麦抗源Sw92对叶锈菌小种PHT的抗性系由一对隐性基因所控制。采用简单重复序列(SSR)技术对Sw92携带的抗性基因进行分子标记,共筛选了371对SSR引物,获得2个引物(WMC494、WMC737)可在抗/感池和双亲中扩增出多态性DNA片段。遗传连锁分析结果表明,该抗病基因位于小麦6BS上,与WMC494、WMC737标记的遗传距离分别为3.4cM和15.0cM,不同于6BS上的已知抗叶锈基因Lr36和Lr53,暂命名为LrSw92。 相似文献
8.
小麦抗叶锈基因Lr19来源于长穗偃麦草,表现优良的抗叶锈性,国内外发现对Lr19有毒性的小麦叶锈菌菌株的报道较少。本研究以TcLr19和Thatcher亲本以及TcLr19×Thatcher F2代单株构建的分离群体为材料,建立了与Lr19共分离的稳定的SCAR分子标记,命名为Y19SCAR982。对49个小麦抗叶锈近等基因系材料的稳定性检测结果表明,其重复性好且为Lr19特异的分子标记。对120个小麦品种检测的结果表明,该标记可有效应用于小麦抗叶锈分子辅助选择育种。 相似文献
9.
小簇麦易位系V9128-3抗条锈病基因的遗传分析和SSR分子标记 总被引：2，自引：0，他引：2
用7个我国当前流行的条锈菌生理小种对V9128-3的抗条锈性进行了评价,表明本易位系对我国优势流行小种具有良好的抗病性。以Su-4对V9128-3与铭贤169配置的F1、BC1F1、F2及F3代群体进行了遗传分析,并对其中1个F2群体进行了SSR标记,再用BC1F1群体的部分单株和F3家系进行连锁标记的初步验证。遗传分析表明了V9128-3对Su-4的抗病性由1对显性核基因独立控制,从219对SSR引物中筛选到2个位于2AL上的该基因YrHV(暂命名)两侧的标记Xgwm356和Xwmc658,遗传距离分别为8.5和5.6cM,所用部分BC1F1单株和F3家系验证了该2个标记与YrHV连锁性。将此标记可用于小麦抗条锈病分子标记辅助育种。 相似文献
10.
中国小麦贵州98-18中抗叶锈基因的分子定位 总被引：2，自引：0，他引：2
小麦(Triticum aestivum)品系贵州98-18对中国目前大多数叶锈菌(Puccinia triticina)生理小种表现抗性。基因推导表明,贵州98-18可能携带新的抗叶锈基因。为了有效利用这一抗源,将贵州98-18和感病小麦品种郑州5389杂交,获得F1、F2代群体,用我国叶锈菌优势小种THTT对双亲及其杂交后代进行接种鉴定。结果表明,贵州98-18对THTT的抗性由1对显性基因控制,暂命名为LrG98。采用SSR技术对贵州98-18携带的抗病基因进行分子标记,共筛选了1 274对SSR或STS引物,位于1BL染色体上的4对引物可在抗/感池和双亲中扩增出多态性DNA片段。遗传连锁分析结果表明,该抗病基因位于小麦1BL染色体上,与Xbarc582-1B和Lr26的STS标记ω-secali(Glu-B3)的遗传距离最近,均为3.8 cM。该基因与目前所有已知的抗叶锈基因不同,可能是1个新的抗病基因。 相似文献
11.
香蕉枯萎病菌生理小种鉴定及其SCAR标记 总被引：8，自引：0，他引：8
通过室内人工接种蕉类鉴别寄主,对采集于广东蕉区的18个蕉类枯萎病菌菌株进行鉴定,KP021、KP022、GZ981和JL021 4个菌株属Racel,其余14个菌株属Race4,说明广东蕉区同时存在尖孢镰刀菌古巴专化型Race1和Race4。用RAPD技术对上述18个菌株进行分析,从200条随机引物中筛选出8条引物可产生生理小种RAPD标记12个,其中标记Racel的8个,标记Race4的4个。对这些RAPD标记带分别进行回收、克隆、测序,根据这些特异片段序列分别设计相应的SCAR引物,通过对18个菌株的PCR扩增检验,有4个RAPD标记成功地转化为SCAR标记,其中Race1-SCAR标记1个、Race4-SCAR标记2个、同时能鉴定出2个小种的SCAR标记1个。应用这4个SCAR标记同时对采自田间的9个病菌分离物进行检测,能够准确地鉴定出广东蕉区的尖孢镰刀菌古巴专化型Racel和Race4,这为下一步开展香蕉枯萎病菌生理小种的分子鉴定及各生理小种田间流行动态监测奠定了基础。 相似文献
12.
ABSTRACT Specific primers and polymerase chain reaction (PCR) assays that identify Fusarium oxysporum f. sp. ciceris and each of the F. oxysporum f. sp. ciceris pathogenic races 0, 1A, 5, and 6 were developed. F. oxysporum f. sp. ciceris- and race-specific random amplified polymorphic DNA (RAPD) markers identified in a previous study were cloned and sequenced, and sequence characterized amplified region (SCAR) primers for specific PCR were developed. Each cloned RAPD marker was characterized by Southern hybridization analysis of Eco RI-digested genomic DNA of a subset of F. oxysporum f. sp. ciceris and nonpathogenic F. oxysporum isolates. All except two cloned RAPD markers consisted of DNA sequences that were found highly repetitive in the genome of all F. oxysporum f. sp. ciceris races. F. oxysporum f. sp. ciceris isolates representing eight reported races from a wide geographic range, nonpathogenic F. oxysporum isolates, isolates of F. oxysporum f. spp. lycopersici, melonis, niveum, phaseoli, and pisi, and isolates of 47 different Fusarium spp. were tested using the SCAR markers developed. The specific primer pairs amplified a single 1,503-bp product from all F. oxysporum f. sp. ciceris isolates; and single 900- and 1,000-bp products were selectively amplified from race 0 and race 6 isolates, respectively. The specificity of these amplifications was confirmed by hybridization analysis of the PCR products. A race 5-specific identification assay was developed using a touchdown-PCR procedure. A joint use of race 0- and race 6-specific SCAR primers in a single-PCR reaction together with a PCR assay using the race 6-specific primer pair correctly identified race 1A isolates for which no RAPD marker had been found previously. All the PCR assays described herein detected up to 0.1 ng of fungal genomic DNA. The specific SCAR primers and PCR assays developed in this study clearly identify and differentiate isolates of F. oxysporum f. sp. ciceris and of each of its pathogenic races 0, 1A, 5, and 6. 相似文献
13.
玉米大斑病菌是异宗配合真菌，有性杂交有可能增强病菌的致病力，或形成新的致病小种，因此对该病菌有性杂交后代进行致病性测定和遗传多态性分析对控制该病菌的危害具有重要意义。对亲本菌株132、135和它们杂交产生的70个单子囊孢子F1代菌株进行了生理小种鉴定和AFLP（扩增性片段长度多态性）分析。生理小种鉴定结果表明，F1代菌株中与亲本菌株132（23N号小种）属于同一小种类型的占41．4％，与亲本菌株135（23号小种）相同的占20．0％，另外还出现了0、1、2、3、13、123、12N、13N和123N号小种，所占比例分别为2．9％、1．4％、2．9％、2．9％、4．3％、8．6％、1．4％、4．3％和10．0％，说明有性杂交可使后代菌株的致病性发生比较广泛的变异。AFLP分析表明，F1代菌株之间分子遗传相似系数在0．87～0．99之间，其中84．3％的F1代菌株与亲本菌株的遗传相似系数在0．878以上，但与亲本菌株132同源性较强的F1代菌株数目大约是与亲本菌株135的5倍，说明不同菌株具有不同的遗传传递能力。比较生理小种鉴定和AFLP分析结果，发现生理小种分化和AFLP分子遗传多态性间有一定的相关性，但不能完全对应，不存在遗传谱系就等于小种的简单对应关系。 相似文献
14.
Downy mildew is a destructive disease of spinach worldwide. There have been 10 races described since 1824, six of which have been identified in the past 10 years. Race identification is based on qualitative disease reactions on a set of diverse host differentials which include open-pollinated cultivars, contemporary hybrid cultivars, and older hybrid cultivars that are no longer produced. The development of a set of near-isogenic open-pollinated spinach lines (NILs), having different resistance loci in a susceptible and otherwise common genetic background, would facilitate identification of races of the downy mildew pathogen, provide a tool to better understand the genetics of resistance, and expedite the development of molecular markers linked to these disease resistance loci. To achieve this objective, the spinach cv. Viroflay, susceptible to race 6 of Peronospora farinosa f. sp. spinaciae, was used as the recurrent susceptible parent in crosses with the hybrid spinach cv. Lion, resistant to race 6. Resistant F(1) progeny were subsequently backcrossed to Viroflay four times with selection for race 6 resistance each time. Analysis of the segregation data showed that resistance was controlled by a single dominant gene, and the resistance locus was designated Pfs-1. By bulk segregant analysis, an amplified fragment length polymorphism (AFLP) marker (E-ACT/M-CTG) linked to Pfs-1 was identified and used to develop a co-dominant Sequence characterized amplified region (SCAR) marker. This SCAR marker, designated Dm-1, was closely linked ( approximately 1.7 cM) to the Pfs-1 locus and could discriminate among spinach genotypes that were homozygous resistant (Pfs-1Pfs-1), heterozygous resistant (Pfs-1pfs-1), or homozygous susceptible (pfs-1pfs-1) to race 6 within the original mapping population. Evaluation of a wide range of commercial spinach lines outside of the mapping population indicated that Dm-1 could effectively identify Pfs-1 resistant genotypes; the Dm-1 marker correctly predicted the disease resistance phenotype in 120 out of 123 lines tested. In addition, the NIL containing the Pfs-1 locus (Pfs-1Pfs-1) was resistant to multiple races of the downy mildew pathogen indicating Pfs-1 locus may contain a cluster of resistance genes. 相似文献
15.
Support for a stepwise mutation model for pathogen evolution in Australasian Puccinia striiformis f.sp. tritici by use of molecular markers 总被引：7，自引：0，他引：7
Since its initial detection in Australia in 1979, wheat yellow (stripe) rust ( Puccinia striiformis f.sp. tritici ) has evolved in Australia and New Zealand into more than 20 pathotypes with assorted virulence characteristics. This evolution is believed to have occurred in a stepwise fashion from an original single pathotype, with no subsequent new introductions. A combination of random amplified polymorphic DNA (RAPDs) and amplified fragment length polymorphisms (AFLPs) was used to examine the level of molecular variation in Australian and New Zealand isolates, and to compare this with variation amongst other isolates of P. striiformis . Using 60 RAPD primers on seven Australian isolates representing seven different pathotypes collected between 1979 and 1991, more than 300 potentially polymorphic loci were analysed and no polymorphisms were detected. Using the same primers on two UK isolates, 3% of loci showed a polymorphism. A similar level of polymorphism was found between UK isolates using AFLP primers, and between 5 and 15% of fragments were polymorphic between an isolate from the UK, an isolate from Denmark, and one from Colombia. However, no AFLP polymorphisms were found amongst 14 Australian and New Zealand isolates tested, at over 100 potentially polymorphic loci. The lack of molecular variation in the Australian and New Zealand collection is consistent with the stepwise mutation theory of pathotype evolution from a single introduction. 相似文献
16.
ABSTRACT Two strains of the wheat stem rust fungus, Puccinia graminis f. sp. tritici, were crossed on barberry, and a single F(1) progeny strain was selfed. The parents, F(1), and 81 F(2) progeny were examined for virulence phenotypes on wheat differential cultivars carrying stem rust resistance (Sr) genes. For eight Sr differentials, phenotypic ratios are suggestive of single dominant avirulence genes AvrT6, AvrT8a, AvrT9a, AvrT10, AvrT21, AvrT28, AvrT30, and AvrTU. Avirulence on the Sr; (Sr 'fleck') differential showed phenotypic ratios of approximately 15:1, indicating epistatic interaction of two genes dominant for avirulence. Avirulence on Sr9d favored a 3:13 over a 1:3 ratio, possibly indicating two segregating genes-one dominant for avirulence and one dominant for avirulence inhibition. Linkage analysis of eight single dominant avirulence genes and 970 DNA markers identified DNA markers linked to each of these avirulence genes. The closest linkages between AvrT genes and DNA markers were between AvrT6 and the random amplified polymorphic DNA marker crl34-155 (6 centimorgans [cM]) AvrT8a and the amplified fragment length polymorphism marker eAC/mCT-197 (6 cM) and between AvrT9a and the amplified fragment length polymorphism marker eAC/mCT-184 (6 cM). AvrT10 and AvrTU are linked at distance of 9 cM. 相似文献
17.
ABSTRACT Collections of Puccinia triticina, the wheat leaf rust fungus, were obtained from Great Britain, Slovakia, Israel, Germany, Australia, Italy, Spain, Hungary, South Africa, Uruguay, New Zealand, Brazil, Pakistan, Nepal, and eastern and western Canada. All single-uredinial isolates derived from the collections were tested for virulence polymorphism on 22 Thatcher wheat lines that are near-isogenic for leaf rust resistance genes. Based on virulence phenotype, selected isolates were also tested for randomly amplified polymorphic DNA (RAPD) using 11 primers. The national collections were placed into 11 groups based on previously established epidemiological zones. Among the 131 single-uredinial isolates, 105 virulence phenotypes and 82 RAPD phenotypes were described. In a modified analysis of variance, 26% of the virulence variation was due to differences in isolates between groups, with the remainder attributable to differences within groups. Of the RAPD variation, 36% was due to differences in isolates between groups. Clustering based on the average virulence distance (simple distance coefficient) within and between groups resulted in eight groups that differed significantly. Collections from Australia-New Zealand, Spain, Italy, and Britain did not differ significantly for virulence. Clustering of RAPD marker differences (1 - Dice coefficient) distinguished nine groups that differed significantly. Collections from Spain and Italy did not differ significantly for RAPD variation, neither did collections from western Canada and South America. Groups of isolates distinguished by avirulent/virulent infection types to wheat lines with resistance genes Lr1, Lr2a, Lr2c, and Lr3 also differed significantly for RAPD distance, showing a general relationship between virulence and RAPD phenotype. The results indicated that on a worldwide level collections of P. triticina differ for virulence and molecular backgrounds. 相似文献
18.
Genetic variation among asexual progeny of Phytophthora infestans detected with RAPD and AFLP markers 总被引：6，自引：1，他引：6
Genotypic variation among 32 single-zoospore isolates (SZI) of Phytophthora infestans , derived asexually from two hyphal-tip parental isolates (PI-105 and PI-1) of the US-8 genotype, was assessed with 80 random amplified polymorphic DNA (RAPD) primers and 18 amplified fragment length polymorphic DNA (AFLP) primer pairs. In previous investigations, the SZIs from parental isolate PI-105 showed high levels of virulence variability and were differentiated into 14 races, whereas the SZIs from PI-1 showed identical virulence to the parent. The purpose of this investigation was to determine if phenotypic variation observed among SZIs of P. infestans could be detected at the DNA level in these isolates. Polymorphism was detected with 51 RAPD primers and with all 18 AFLP primer pairs in PI-105 SZIs. In SZIs from PI-1, polymorphism was also detected with 25 RAPD primers and 17 AFLP primer pairs. Cluster analysis using the unweighted pair-group method with arithmetic averages (UPGMA) separated the SZIs from parent PI-105 into six virulence groups, 11 RAPD groups and three AFLP groups. Cluster analysis of PI-1 SZIs, which all belong to the same virulence group, differentiated them into four RAPD groups and six AFLP groups. No close correlation among RAPD, AFLP and virulence groups could be established within the two progenies of SZIs. Results of this study suggest that there is a considerable level of inherent genetic variability among SZIs derived asexually from the same parental isolate. The possible mechanisms and implications of this genetic variation are discussed. 相似文献