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摘要:分子标记技术筛选亲本单26与单28的遗传差异条带,并成功转化为单26菌株核DNA的特征标记SCAR1和单28菌株核DNA的特征标记SCAR2;将核特征标记SCAR应用于杂交菌株2628的原生质体再生菌株的检测,成功地分离与鉴定出两亲本类型的原生质单核化菌株,表明杂交菌株2628细胞已被拆分;本研究再通过配对杂交、SCAR检测及出菇试验等证明,两亲本类型的原生质单核化菌株又可再重建成异核体的2628菌株;这为丰富食用菌特别是同宗结合真菌的遗传育种理论研究提供了新的思路。 相似文献
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烟草PVY抗性的遗传分析与分子标记筛选 总被引:2,自引:0,他引:2
本文以抗马铃薯Y病毒(简称PVY)的烟草品种RY5,感病品种Coker176为亲本,构建F1、正反交F2和正反交BC1群体,苗期摩擦接种PVY的抗性遗传分析结果表明,接种后第21d群体PVY抗性数据符合孟德尔单基因隐性质量性状的遗传模型。接种后第28d群体PVY抗性数据偏离孟德尔单基因隐性质量性状的遗传模型。提取F2代群体中抗病和感病单株DNA,从多条RAPD引物和一对SCAR引物中,筛选出两个紧密连锁的分子标记。RAPD标记O12V3695与RY5的抗病基因对应的显性等位基因位点(Va)间的遗传距离为2.10cM,而SCAR标记与Va间的遗传距离为2.52cM,这两个分子标记可用于抗PVY抗性育种。 相似文献
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草菇杂交菌株2628的鉴定与品比试验 总被引:1,自引:0,他引:1
将2个匍匐型不孕的草菇单孢菌株单26、单28进行杂交配对,尖端菌丝细胞分离纯化后得到一气生型菌株2628。RAPD标记和SRAP标记的DNA指纹表明,2628继承包含了单26和单28的所有遗传谱带,是一株杂交种。杂交菌株2628可出菇,在与当前主栽品种出菇品比试验中,杂交菌株2628子实体外观较好,产量最高,是主栽品种V23-2产量的2倍多,生物学转化率达到33.4%。 相似文献
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结球甘蓝霜霉病是由十字花科寄生霜霉菌引发的严重病害。本研究利用高代自交系‘R103’抗病亲本和‘S101’感病亲本及其F2分离群体,于霜霉病病发高峰期田间自然诱发抗性鉴定,由单基因显性控制。运用AFLP分子标记技术,结合BSA法,对双亲和2对抗、感基因池的筛选和验证,获得2个与结球甘蓝抗霜霉病基因相关的AFLP标记。204 bp BoRAAC/CAC标记转化为SCAR标记,发现在抗病亲本和抗病池中与感病亲本和感病池中条带只存在量上的差异,在抗性选择起中只能起辅助作用;191 bp BoRAAG/CTC标记(EU-635443.1),转化为SCAR标记,仅在抗病亲本和2个抗病池中获得目的条带,经F2群体单株验证后,与抗性位点的遗传距离为5.7cM。利用BoRAAG/CTC113标记对F1和BC1群体与多年苗期霜霉病抗性鉴定的20份结球甘蓝种质资源进行验证和筛选,该标记与品种(系)的霜霉病抗性吻合率达到95%,初步表明该标记可应用于早期结球甘蓝抗霜霉病抗性辅助选择。 相似文献
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利用分子标记辅助育种技术选育低温高产草菇菌株 总被引:1,自引:0,他引:1
《分子植物育种》2017,(8)
利用RAPD分子标记技术对11个草菇菌株进行了遗传背景分析,发现泰国No.1菌株与低温菌株VH3的亲缘关系较远,可作为杂交育种的另一亲本菌株。收集两亲本的孢子并进行单孢杂交配对,采用低温筛选试验淘汰掉部分不耐低温的菌株,随后利用交配型基因分子标记验证杂交子的真实性,最终共获得77个杂交子。将部分杂交菌株进行低温出菇栽培试验,结果显示:草菇杂交菌株V6T3-1的子实体农艺性状较好,且生物转化率达到28.26%,远高于亲本菌株VH3与泰国No.1。 相似文献
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大豆新品系已成为杂交育种中最主要亲本类型,本研究对系谱明确、适合江淮地区种植的296份大豆新品系进行SSR和PAV标记分析,以揭示其遗传关系,促进其育种利用。结果 93个SSR标记共检测到417个等位变异,平均每个位点等位变异数为4.48,变幅为2~15,PIC值为0.46;227对PAV标记每个位点平均等位变异数为2.10,变幅2~4,PIC值为0.22;基于SSR标记所计算的多样性指标数值均高于PAV标记所得。根据核心亲本划分的4个亚群间分子标记遗传多样性值相近,但都存在一些特有和特缺等位变异。基于SSR和PAV标记遗传距离的聚类分析分别可将供试材料分为12和10类,其中4个大类均可与4个核心亲本亚群对应,还发现一些系谱相同/相近的品系被聚在不同类群。两类分子标记都可用于揭示供试材料的遗传背景,利用所有320个标记可将296份新品系分为8类。 相似文献
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用一套分别含有不同抗叶锈基因的53个以Thatcher为遗传背景的近等基因系(near-isogeniclines,NILs)对已报道的分别与抗叶锈基因Lr24和Lr35连锁的STS、SCAR进行特异性验证。结果对于与Lr24连锁的STS标记,在53个NILs中只在TcLr24亲本中扩增出片段大小与报道相同的310bp的条带,在TcLr35中也扩增出了一条片段,但片段大小不同于310bp约为270bp。对于与Lr35连锁的SCAR标记,只在TcLr35亲本中扩增出片段大小为900bp的条带,与报道片段大小一致。验证结果表明与抗病基因Lr24和Lr35连锁的STS、SCAR分子标记在NILs中特异性都较好,进一步证明了这两个分子标记可方便地用于小麦抗叶锈基因Lr24、Lr35的分子标记辅助选择育种。 相似文献
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以甘薯抗茎线虫病品种徐781和感茎线虫病品种徐薯18的杂交F1分离群体的174株单株为材料,对甘薯抗茎线虫病基因的遗传进行了分析。结果表明,徐781的抗茎线虫病为单基因控制,推测其基因型为Rrrrrr,徐薯18的基因型为rrrrrr。采用BSA-RAPD相结合的方法,筛选940条随机引物,发现10条引物在抗、感池间表现多态性。用这10条引物检测两亲本及建池单株,发现只有引物OPP03在8株抗池单株中扩增出一条在8株感池单株中所没有的特异条带,认为该标记与甘薯抗茎线虫病基因连锁。根据该标记对F1代174个单株的扩增结果,利用Mapmaker3.0软件计算遗传距离,表明该标记与抗茎线虫病基因间的遗传距离为14.2cM。将该片断回收、克隆、测序,表明其长度为878bp,该标记命名为OPP03878。根据测序结果,设计1对特异引物,进行特异性扩增,成功地将OPP03878标记转化为SCAR标记,用亲本及分离群体验证表明该分子标记稳定性好。 相似文献
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Rs1046AB is a dominant genic male sterility (DGMS) line in rapeseed, in which the sterility has always been thought to be conditioned by the interaction of a male sterility gene ( Ms ) and its non-allelic restorer gene ( Rf ). This system provides not only a tool for assisting in recurrent selection but also a promising system for hybrid production. Based on previous studies, two amplified fragment length polymorphism markers linked with the Ms gene were converted into a dominant and a co-dominant sequence characterized amplified region (SCAR) marker, respectively. The putative linear order relationship of three dominant SCAR markers with the same genetic distance from the Rf gene, was also determined by an examination of whether the homologues of these markers are present or not in different lines carrying Rf . A bigger fragment generated by the closest marker linked to the Rf gene was observed in all lines carrying the recessive allele rf , suggesting that this marker is a co-dominant marker, which was further confirmed by nucleotide sequence comparison of these fragments. SCAR markers specific for Ms and Rf will be especially valuable in marker-assisted DGMS three-line breeding. 相似文献
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Rye (Secale cereale L. and S. strictum) offers potential to increase the genetic variability and to introduce desirable characters for wheat improvements. Cytogenetic
techniques have been used to screen wheat lines containing rye chromatin. These techniques are not adequate since they are
highly technical and time consuming. They are not suitable for breeding programs that require rapid screening of large numbers
of genotypes. The main objective of this study was to develop and characterize ISSR and SCAR markers that can distinguish
wheat from rye genome. Total DNA from wheat, rye, and triticale accessions from different provenances were amplified with
ISSR primers in PCR assays. Three wheat-diagnostic sequences were identified. In addition three rye-diagnostic ISSR markers
of which, one marker specifically diagnostic for Secale strictum were characterized. Pairs of primers flanking these specific sequences were designed to produce SCAR markers. Two SCAR markers
were rye genome-specific. One SCAR was present in all the seven rye chromosome, and another was specific to rye chromosomes
two, three, four, and seven. These newly developed ISSR and SCAR markers should be useful to wheat breeders screening genotypes
that may contain rye chromatins. 相似文献
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Rs1046AB is a line which is true breeding for a dominant genetic male sterility gene (Ms) but which is a mixture of male fertile and sterile individuals (a two-type line) because it is segregating for a dominant suppressor gene (Rf). This system provides a promising alternative to the CMS system for hybrid breeding in Brassica napus. In order to identify molecular markers linked to the rf gene, a near-isogenic line (NIL) population from the cross between a sterile individual (MsMsrfrf) and a fertile individual (MsMsRfrf) in Rs1046AB was subjected to amplified fragment length polymorphism (AFLP) analysis, with a combination of comparing near isogenic lines (NILs) and bulked segregant analysis (BSA). From 2,816 pairs of AFLP primers, six fragments showing polymorphism between the fertile and sterile bulks as well as the individuals of the bulks were identified. Linkage analysis indicated that the six AFLP markers are tightly linked to the Rf gene and all are distributed on the same side. The minimum genetic distance between the Rf gene and a marker was 0.7 cM. Since the AFLP markers are not suitable for large-scale application in MAS (marker-assisted selection), our objective was to develop a fast, cheap and reliable PCR-based assay. Consequently, three of the four closest AFLP markers were converted directly to sequence characterized amplified region (SCAR) markers. For the other marker a corresponding SCAR marker was successfully obtained after isolating the adjacent sequences by PCR Walking. The available SCAR markers of the Rf gene will greatly facilitate future breeding programs using dominant GMS to produce hybrid varieties. 相似文献
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Yan Yan Yang Yu Meng Huo Jun Miao Bing Jiang Liu Su Ping Kong Li Min Gao Chang Liu Zhen Bao Wang Yasuki Tahara Hidemi Kitano Xiong Wu 《Euphytica》2013,190(2):267-277
Cytoplasmic genetic male-sterility is used to produce hybrid onion (Allium cepa L.) seeds worldwide. In this paper, we present the results of research aimed toward identifying PCR-based markers linked to the Ms locus through amplified fragment length polymorphism (AFLP). After screening 512 AFLP primer combinations, only one AFLP fragment was identified as being flanking linked to the dominant Ms allele. Subsequently, the AFLP marker was converted into a sequence-characterized amplified region (SCAR) marker, designated as DNF-566, co-segregated with the dominant Ms allele in first backcross (BC1) segregated populations. Furthermore, we designed another molecular marker (RNS-357) co-segregated with the ms allele to identify different genotypes (i.e., MsMs, Msms, or msms). Both markers could be used for evaluating onion lines with different genetic backgrounds (including male-sterile lines, maintainer lines, male-fertile lines, and commercial based F1 hybrid cultivars). The results of this study indicate that maintainer plants could be directly selected by using these 2 SCAR markers in the onion breeding process, and this may contribute significantly toward breeding onion F1 hybrid cultivars. 相似文献
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There is an urgent need for the developmentof early identification techniques inolive-trees due to the economic importanceof cultivar identification in periods ofexpansion like now. We have been able toidentify 22 olive-tree cultivars using only10 different, specific, repeatable markers.These markers were designed by the cloningof significant RAPD bands obtained in PCRperformed on bulked DNA to retain thegenetic variability of each cultivar.Clones were partially or totally sequencedand new primers derived from thesesequences were used to obtain SequenceCharacterised Amplified Region (SCAR)fragments. We have demonstrated that theuse of the 10 SCAR markers is enough toprovide a simple, cheap, and reliableprocedure to identify 22 geographicallyrelated olive-tree cultivars. 相似文献
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Jatropha curcas (Euphorbiaceae) is an oil-bearing species with multiple uses and considerable potential as a bioenergy crop. The present
investigation has been undertaken to assess the extent of genetic diversity in a representative set of 42 accessions of J. curcas encompassing different crop growing regions in India along with a non-toxic genotype from Mexico as a prelude for utilization
of promising and genetically divergent materials in the breeding programmes. Molecular polymorphism was 42.0% with 400 RAPD
primers and 33.5% with 100 ISSR primers between accessions indicating modest levels of genetic variation in the Indian germplasm.
The within-population variation based on RAPD polymorphism was 64.0% and was on par with the inter-population variation. Polymorphic
ISSR markers have been identified that could differentiate the Indian accessions from the Mexican genotype and two of them
were converted to SCAR markers. The SCAR primer pair ISPJ1 amplified a 543 bp fragment in all the Indian populations, while
ISPJ2 with a specific amplicon of 1,096 bp was specific to the Mexican genotype. Population-specific bands have been identified
for the accession from Kerala (2 RAPD markers), Neemuch-1 from Rajasthan (1 each of RAPD and ISSR markers) and the non-toxic
genotype from Mexico (17 RAPD and 4 ISSR markers), which serve as diagnostic markers in genotyping. The study indicates an
immediate need for widening the genetic base of J. curcas germplasm through introduction of accessions with broader geographical background. 相似文献
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多重PCR技术鉴定番茄Ty-1和Mi基因 总被引:2,自引:0,他引:2
利用同一PCR反应体系,对分别与番茄(Lycopersion esculentum)抗黄化曲叶病毒(Tomato YellowLeaf Curl virus)病的Ty-1基因和番茄抗根结线虫(root-knot nematode)的Mi基因紧密连锁的SCAR标记进行同时扩增筛选,扩增的特异性片段与单引物扩增片段完全吻合.与Ty-1基因紧密连锁的SCAR1标记为共显性标记,抗感材料均产生398 bp的特异片段,纯合和杂合抗病基因型存在TaqⅠ酶切位点,酶切后分别产生了303 bp和95 bp以及398 bp、303 bp和95 bp的特异性片段,而感病基因型无此酶切位点.与Mi基因紧密连锁的SCAR2标记也为共显性标记,抗感材料均产生750 bp的特异片段,纯合和杂合抗病基因型存在TaqⅠ酶切位点,酶切后分别产生了570bp和180bp以及750bp、570bp和180bp的特异性片段,而感病基因型无此酶切位点.酶切结果仍为750 bp的产物.经反复验证,结果准确可靠,可以用于在同一PCR反应体系中对两个抗病基因进行同时筛选鉴定. 相似文献