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1.
To test if locus-specific microsatellite markers designed for one genus are informative when used with related genera, the conservation of microsatellite-flanking intergeneric primer binding sites was tested in the closely related tribes Vicieae and Cicereae, from the subfamily Papilionoideae of the Leguminosae family. A total of 123 sequence-tagged microsatellite sites (STMS) markers derived from chickpea were used to amplify loci in lentil (Lens) and dry pea (Pisum). The percentage of chickpea primer binding sites conserved between the three genera was 54.4%. Hybridisation of 63 selected amplified loci to the digoxigenin-labelled oligonucleotide probe (TAA)5 showed that 69.8% of loci from dry pea and 66.6% of loci from lentil hybridised to the probe. Sequencing of amplified products from chickpea with the primer Ta176 demonstrated that one amplicon contained a microsatellite, whereas another amplicon amplified with the same particular STMS primer pair did not. Amplicons produced from lentil and pea with this primer pairs did not contain microsatellite sequences. Results obtained with Tr7, which amplified a PCR product in lentil and chickpea but not in pea, showed that microsatellite sequences were present in chickpea and absent in lentil. Similar results were obtained with Ts35, which produces amplicons in pea and chickpea; but, again, microsatellite sequences were only present in chickpea. We therefore conclude that STMS derived from chickpea could be used to detect variability between other Leguminosae genera, but it is necessary to verify whether homologous loci are revealed. 相似文献
2.
The potential of microsatellite markers for use in genetic studies in eggplant, Solanum melongena, has been evaluated. A genomic library of eggplant was screened for GA and GT repeat motifs to isolate microsatellite clones. The frequency of each repeat motif in the eggplant genome was found to be every 3200 kb for GA repeats and every 820 kb for GT repeats. Sixty‐one per cent of GT repeats were found to directly flank AT repeats. A total of 37 polymerase chain reaction (PCR) primer pairs were designed, 23 of which amplified a single product or several products. The level of microsatellite polymorphism was evaluated by using S. melongena lines and related Solanum species. Two to six alleles per primer pair were displayed in the S. melongena lines and two to 13 alleles were displayed in the Solanum relatives. Seven microsatellites showed polymorphism between parental lines of the mapping population and segregated in a codominant Mendelian manner. These microsatellite loci were distributed throughout the linkage map. 相似文献
3.
Yufang Guo Sukumar Saha John Z. Yu Johnie N. Jenkins Russell J. Kohel Brian E. Scheffler David M. Stelly 《Euphytica》2008,161(3):361-370
Bacterial artificial chromosome (BAC) libraries with large DNA fragment inserts have rapidly become the preferred choice for
physical mapping. BAC-derived microsatellite or simple sequence repeats (SSRs) markers facilitate the integration of physical
maps with genetic maps. The objective of this research was to identify chromosome locations of the BAC-derived SSR markers
in tetraploid cotton. A total of 192 SSR primer pairs were derived from BAC clones of an Upland cotton genetic standard line
TM-1 (Gossypium hirsutum L.). Metaphor agarose gel electrophoresis results revealed 76 and 59 polymorphic markers between TM-1 and 3–79 (G. barbadense) or G. tomentosum, respectively. Using deletion analysis method, we assigned 39 markers out of the 192 primer pairs to 17 different chromosomes
or chromosome arms. Among them, 19 and 17 markers were localized to A-subgenomes (chromosome 1–13) and D-subgenomes (chromosome
14–26), respectively. The subgenome status for the remaining three markers remained unclear due to their two potential chromosome
locations achieved by tertiary monosomic stocks deletion analysis. Chromosomal assignment of these BAC-derived SSR markers
will help in integrating physical and cotton genetic linkage maps and thus facilitate positional candidate gene cloning, comparative
genome analysis, and the coordination of chromosome-based genome sequencing project in cotton.
Disclaimer: Mention of trademark, proprietary product, or vendor does not constitute a guarantee or warranty of the product
by USDA, ARS and does not imply its approval to the exclusion of other products or vendors that may also be suitable.
The U.S. Government’s right to retain a non-exclusive, royalty-free license in and to any copyright is acknowledged. 相似文献
4.
5.
用227对小麦微卫星引物进行PCR扩增，76对可在多枝赖草和耐黄矮病的普通小麦-多枝赖草二体异附加系Line24的小麦亲本中国春、丰抗13间检测到多态性。和多枝赖草相同而与Line24其他小麦亲本不同的扩增带。在这76对引物中，发现有4对引物能从Line24中扩增出进一步用Line24和普通小麦杂交得到的7个不同的单体异附加系进行验证，也得到同样的结果，说明这4对微卫星引物扩增出的特异带可以作为Line24中多枝赖草染色体的分子标记。根据这4对引物各自对应的微卫星标记位点在小麦染色体上的位置，说明Line24中附加的一对多枝赖草染色体是第3，5，6和7部分同源群多枝赖草染色体相互易位形成的。 相似文献
6.
Abundance, polymorphism and genetic mapping of microsatellites in oilseed rape (Brassica napus L.) 总被引：9，自引：0，他引：9
The objective of the present study was to estimate the abundance and degree of polymorphism of simple sequence repeat (SSR) markers in rapeseed. By screening about 45000 clones of a small inserts library of rapeseed total DNA the abundances of GA/TC and CA/TG simple sequence repeats in the rapeseed genome were estimated to be approximately one repeat every 100 kb and 400 kb, respectively. After sequencing 13 positive clones, primer pairs could be designed for 11 microsatellite loci. Seven of these primer pairs produced reproducible amplification products in a set of 31 rapeseed genotypes, with one pair amplifying two independent products, giving a total of eight amplified loci. The different microsatellite loci displayed between one and three visible alleles. At four loci, additional null alleles were observed. With up to four alleles, polymorphic microsatellite markers show significantly higher allele numbers in rapeseed than restriction fragment length polymorphism (RFLP) markers. Four of the eight microsatellite markers could be mapped on four different linkage groups of an RFLP map of the rapeseed genome. 相似文献
7.
为了揭示人工甘蓝型油菜早期世代遗传和表观遗传变异规律, 以A组合(大黄油菜×中花芥蓝) S0世代、B组合(大黄油菜×中迟芥蓝) S0和S1世代人工甘蓝型油菜为材料, 分别利用AFLP和MSAP技术检测基因组变化及甲基化模式变化情况。结果表明, 16对引物在A组合S0扩增到523条带, 其中4对引物扩增出9条变异带, 包括7条亲本缺失带和2条新增带, 分别占S0总条带的1.33%和0.38%;45对引物在B组合双亲植株扩增到1093条带, 只有1对引物检测到1条父本带型在所有S0植株中缺失, 约占S0总条带的0.09%;在B9子代F19-1~F19-16总共扩增得到1092条带, 变异带有10条, 占总条带的0.915%, 其中包括9条缺失带和1条新增带, 9条缺失带全部位于C基因组。MSAP检测发现, B组合S0植株中有3个位点发生了甲基化模式的改变, 全部位于A基因组, 甲基化模式改变位点占总检测位点的1.37%。研究还发现B组合S0世代一个植株出现可遗传的花色变异, 推测该表型变异与B组合人工甘蓝型油菜中C基因组变异有关。 相似文献
8.
保守性强、重复性好、多态性高的微卫星位点可被有效用于构建作物DNA条形码。选取目前生产上主要推广种植、代表不同来源系统的12个棉花品种作为微卫星位点筛选材料,参考我室构建的四倍体栽培棉种种间高密度遗传图谱信息,从376对覆盖全基因组的SSR引物中,筛选出51对引物可扩增出带型清晰且多态性高的微卫星位点。这些引物在12个供试品种中共产生155个等位位点,每对引物揭示的等位基因位点在2~7之间,平均值为3.04。参照微卫星位点的染色体定位和多态信息,在每条染色体上选择一个多态性相对高的SSR位点,其相应的26对SSR引物被推荐为构建棉花品种DNA条形码的一套首选引物,并初步应用于12个品种的DNA条形码编制。其余25对引物作为候选引物。使用该套引物扩增出的微卫星位点可用于大量棉花品种DNA条形码构建,为棉花品种真实性和纯度的分子鉴定奠定基础。 相似文献
9.
棉花第14染色体分子标记连锁群的构建及其应用 总被引：2，自引：1，他引：1
利用置换系CSB14Sh与TM-1产生F2分离群体,以SSR标记构建连锁图对置换系进行分子鉴定。利用从覆盖全基因组的3800对引物筛选出的15对多态性引物对群体进行扩增,产生23个分子标记位点。其中,21个分子标记进入了同1个连锁群。通过与前人的分子图谱比较,把该连锁群定位在第14染色体上。表明TM-1与CSB14Sh在第14条染色体上有差异,而在其它染色体上没有差异。再结合染色体置换系构建的过程,可以认为CSB14Sh确实为第14条染色体短臂的置换系。 相似文献
10.
In this study, AFLP and SSR techniques were combined with the bulk segregant analysis (BSA) method to map the restorer gene BrRfp using an F2‐segregating population comprising 258 individuals developed by crossing the polima (pol)‐like cytoplasmic male sterility (CMS) line 06J45 and the restorer line 01S325 of heading Chinese cabbage. A survey of 2048 AFLP primer pairs identified 21 polymorphic fragments, approximately half of which exhibited high similarity with the A09 chromosome sequence of Brassica rapa in the Brassica database (BRAD). Based on the genome sequence, three specific AFLP fragments linked with BrRfp were successfully converted into sequence‐characterized amplified region (SCAR) markers, named SC1233, SC2673 and SC2141. Subsequently, 178 pairs of SSR primers were redesigned for further screening, with five producing polymorphic amplification patterns. Linkage analysis showed that these markers were distributed along both sides of the BrRfp gene, with two markers, SSR03 and SSR2528, co‐segregating with the BrRfp locus in the F2 population. These results may be valuable for marker‐assisted selection and map‐based cloning in heading Chinese cabbage. 相似文献
11.
Identification and DNA polymorphism of the S‐locus receptor kinasegene (SRK) was analysed by pollen tube tests, polymerase chain reaction‐cleaved amplified polymorphic sequence (PCR‐CAPS) and nucleotide sequencing. SRK‐specific primers that can distinguish class and class II S haplotypes amplified single DNA fragments of 900‐1050 bp. The DNA fragments of 22 inbred lines amplified with a class SRK‐specific primer pair determined seven types with HinfI and EcoRII. In addition, the DNA fragments of 17 inbred lines amplified with a class II SRK‐specific primer pair determined three types with Hinf1. Nucleotide sequencing of the DNA fragments amplified from 10S haplotypes showed that exons of the 3′‐end in SRK are highly conserved, and that there is much variation of the introns, which produced polymorphism of the band pattern in PCR‐CAPS profiles. The S haplotypes of the plants were determined by restriction analysis of PCR products and agreed with results based on pollen tube growth tests. The PCR‐CAPS analysis using specific primer pairs of SRK is considered to be useful for S allele identification in breeding programmes. 相似文献
12.
Genomic in situ hybridization (GISH) and restriction fragment length polymorphism (RFLP) were used to identify the Leymus multicaulis (XXNN, 2n = 28) chromosomes in wheat-L. muliticaulis derivatives. Fifteen lines containing L. multicaulis alien chromosomes or chromosomal fragments were identified. All alien chromosomes or fragments in these 15 lines were from
the X genome and none were from the N genome. Eleven L. multicaulis disomic addition lines and four translocation-addition lines were identified with chromosome rearrangements among homoeologous
groups 2, 3, 6 and 7. Only homoeologous group 1 lacked rearrangements in addition or translocation chromosomes. The results
revealed that translocation in non-homoeologous chromosomes widely exists in the Triticeae and therefore it is necessary to
identify the alien chromosomes
(segments) in a wheat background using these combined techniques. During the course of the work, probe PSR112, was found to
detect X genome addition lines involving L. multicaulischromosomes. This may prove to be a valuable probe for the identification of alien chromosomes in a wheat background.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
13.
Jana Řepková Antonín Dreiseitl Pavel Lízal Zdeňka Kyjovská Kateřina Teturová Radka Psotková Ahmed Jahoor 《Euphytica》2006,151(1):23-30
Summary Four newly detected accessions of wild barley (Hordeum
vulgare ssp. spontaneum) resistant to powdery mildew caused by Blumeria
graminis f. sp. hordei were studied with the aim of finding the number of genes/loci conferring the resistance of individual accessions, the type of inheritance of the genes and their relationships to the Mla locus. F2 populations after crosses between the winter variety ‘Tiffany’ and four wild barley accessions and use of microsatellite DNA markers were focused on the identification of individual resistance genes/loci by means of their chromosomal locations. In PI466495, one locus conferring powdery mildew resistance was identified in highly significant linkage with the marker Bmac0213. This location is consistent with the known locus Mla on chromosome 1HS. In the other three accessions the resistance was determined by two independent loci. In PI466197, PI466297 and PI466461, one locus was identified on chromosome 1HS and three new loci were revealed on chromosomes 2HS (highly significant linkage with Bmac0134), 7HS (highly significant linkage with Bmag0021) and 7HL (significant linkage with EBmac0755). Our prospective aim is identification of further linked DNA markers and the exact location of the resistance genes on the barley chromosomes. 相似文献
14.
草棉EST-SSRs的遗传评价 总被引：5，自引：2，他引：3
根据GenBank中公布的247条草棉EST序列,搜索SSR并进行引物设计。其中的25条序列含有27个SSR,1~6碱基重复类型都存在,二碱基和三碱基重复的频率较高。为了明确在A、D和AD基因组中的可转移性,依据25条序列共设计25对EST-SSR引物,其中22对引物扩增出清晰可辨的DNA条带,产生92个多态性片段,平均每对引物产生3.64个多态性片段。引物的多态性信息含量(PIC)在0.49~0.91之间,平均为0.81。6对引物在BC1种间作图群体[(鄂棉22 × Pima3-79) ×鄂棉22]中表现多态性,产生7个多态性位点,其中5个为共显性,2个为显性。除HAU230b标记在BC1分离群体中不符合孟德尔式分离比例,其余引物表现正常分离。6个位点被整合到陆地棉和海岛棉种间BC1遗传连锁图谱上的6条染色体：有4个位于A亚基因组的4条染色体上(Chr.6、10、11和12),2个位于D亚基因组的2条染色体(Chr.19和20)。 相似文献
15.
CSB14Sh,which is isogenic for its recurrent parent TM-1 except for chromosome 14 short arm,was crossed with TM-1,and the F2 population was produced.A total of 3800 SSR primer pairs covering the whole genome were used to screen polymorphism among two parents,TM-1 and CSB14Sh,and their F1 progeny,which resulted in 15 polymorphic primer pairs.The 15 polymorphic primer pairs amplified 23 marker loci. 相似文献
16.
Summary The meiotic pairing behaviour at metaphase I of a Triticum aestivum×Triticum monococcum hybrid has been studied by means of the C-banding technique to ascertain the homology between the chromosomes in the A genome of the two species. The technique allowed the A and B genome chromosomes and the 2D, 3D and 5D chromosomes to be identified. Differences in the level of chromosome pairing in the A genome were noted. The T. monococcum 4A chromosome did not pair with any of the T. aestivum chromosomes in any of the metaphase I cells analysed. Two reciprocal translocations between the 2B and 2D chromosomes on one side and the 2A and 3D on the other side have been identified. The usefulness of the C-banding technique in the study of chromosome homology among species related to wheat is discussed. 相似文献
17.
Efficiency of PCR-RF-SSCP marker production in Brassica oleracea using Brassica EST sequences 总被引：1，自引：0，他引：1
Efficiencies of SCAR, CAPS and PCR-RF-SSCP marker production were investigated using two combinations of breeding lines in Brassica oleracea. Published EST sequences of B. oleracea, Brassica rapa, Brassica napus, and Arabidopsis thaliana and newly determined nucleotide sequences of anther cDNA clones from B. oleracea were used for designing primer pairs to amplify genes. The percentage of primer pairs yielding DNA amplification of a single gene was higher in primer pairs of B. oleracea (91%) than those of B. rapa (56%) and A. thaliana (17%). Single DNA fragments amplified by 9% of the primer pairs showed polymorphism as SCAR markers between a broccoli line and a Chinese kale line by agarose-gel electrophoresis. CAPS analysis showed different band patterns in 32% of the same-sized DNA fragments, and PCR-RF-SSCP analysis revealed DNA polymorphism in 52% of those showing no DNA polymorphism by CAPS. In total, 71% of the single DNA fragments were converted to DNA markers. The frequency of DNA polymorphism between parental lines of a cabbage F1 hybrid was lower, 5% by SCAR and 12% by CAPS. However PCR-RF-SSCP analysis revealed DNA polymorphism in 21% of the DNA fragments showing no polymorphism by CAPS. These results suggest that PCR-RF-SSCP analysis enables highly efficient DNA marker production for mapping of genes in Brassica using progeny, even progeny of closely related parents. Analysis of selfed seeds of broccoli F1 cultivars using PCR-RF-SSCP markers indicated that PCR-RF-SSCP analysis is also applicable to seed purity tests. 相似文献
18.
Isolation of a new repetitive DNA sequence from Secale africanum enables targeting of Secale chromatin in wheat background 总被引：1，自引：0，他引：1
Cheng Liu Zu-Jun Yang Guang-Rong Li Zi-Xian Zeng Yong Zhang Jian-Ping Zhou Zhao-Hui Liu Zheng-Long Ren 《Euphytica》2008,159(1-2):249-258
A genome specific DNA sequence that detects Secale africanum chromatin incorporated into wheat was developed in this study. Random amplified polymorphic DNA (RAPD) analysis was used
to search for genome specific DNA sequences of S. africanum in lines, R111, “mianyang11” (MY11) and wheat-rye 1RS/1BL translocations R25 and R57. A high copy rye-specific DNA segment
pSaD15940 of the S. africanum genome was obtained. The sequence of pSaD15 did not show any significant homology to other reported sequences in databases
and it is therefore a new repetitive sequence of Secale. PCR primers were designed for pSaD15940, which amplify a clear 887 bp fragment in S. africanum but not in any wheat. The primers also amplified an 887 bp fragment in other accessions of rye, Chinese Spring-Imperial rye
chromosome additions and a diverse range of material carrying different rye chromosomes or chromosomal segments. In situ hybridization
showed that probe pSaD15940 was specifically hybridized throughout all rye chromosomes arms except for the terminal regions. The advantage of the rye-specific
probe developed herein compared to those of previous reports is that it has been shown to be widely applicable to other Secale species. The probe will be useful as a molecular marker for the introgression of S. africanum and other rye chromosome segments into the wheat genome. 相似文献
19.
Summary Tetraploid Bromus ciliatus L. is a North American bromegrass that has been placed in the Pnigma section of Bromus. The objective of this study was to characterize the genome of tetraploid B. ciliatus by cytogenetic methods and compare it to the genomes of other species included in the section Pnigma. All the plants of the accession (USDA PI 232214) selected for chromosome counting were tetraploids (2n = 28). The mean 2C nuclear DNA content for tetraploid B. ciliatus was 19.13± 0.07 pg as determined by flow cytometry which is significantly greater than the tetraploid DNA content of B. inermis Leyss. (11.74± 0.16 pg). C-banding procedures were used to identify individual mitotic chromosomes and to develop a karyotype
for B. ciliatus. The genome of the tetraploid B. ciliatus consisted of 16 median chromosomes, eight submedian chromosomes, and four chromosomes with satellites which included one
pair with a large satellite and one pair with a small satellite. The general pattern of the distribution of constitutive heterochromatin
in B. ciliatus was quite different than the other bromegrasses that have been analyzed to date. Except for two pairs of chromosomes, all
chromosomes in tetraploid B. ciliatus had telomeric bands on one or both arms. Some of the chromosomes with telomeric bands had centromeric bands that were located
at one or both sides of the centromere and intercalary bands which were generally absent in the other bromegrass species.
It was possible to identify all chromosomes of tetraploid B. ciliatus and to match the pairs of homologous chromosomes by using chromosome lengths, arm length ratios and C-banding patterns. The
results of this study indicate that tetraploid B. ciliatus has different genomes than the European species evaluated to date in the section Pnigma. 相似文献
20.
Elongated glumes are present in thetetraploid wheat species T.polonicum, T. turanicum, T.durum convar. falcatum and in thehexaploid species T. petropavlovskyi.Inheritance of glume length was studiedwith the aim to map the respective lociusing wheat microsatellite markers. In T. polonicum and T. petropavlovskyiloci conferring long glume were mapped nearthe centromere on chromosome 7A. These twoloci are designated P-A
pol
1 andP-A
pet
1, respectively. It isshown that both are probably homoeoallelicto each other and to the P gene ofT. ispahanicum on chromosome 7B. The loci determining elongated glumes in T. turanicum and T. durum conv. falcatum are not homoeologous to the P loci in the centromeric region of thegroup 7 chromosomes. 相似文献