Characteristics, development and mapping of Gossypium hirsutum derived EST-SSRs in allotetraploid cotton
Characteristics, development and mapping of Gossypium hirsutum derived EST-SSRs in allotetraploid cotton
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DOI：
10.1007/s00122-005-0142-9
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发表时间：
2006-02-01
影响因子：
5.4
通讯作者：
Zhang, TZ
中科院分区：
文献类型：
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作者：
Han, ZG;Wang, CB;Zhang, TZ
为了构建饱和遗传图谱和促进标记辅助选择(MAS)育种，有必要加强目前棉花已知分子标记的储备库。微卫星或简单序列重复(SSR)出现在植物的表达序列标签(EST)中(Kantty等人，Factory Mol Biol 48：501-510,2002)。许多EST现在是公开可用的，是开发EST-SSR的一个很好的工具。从我们为陆地棉构建的两个棉纤维/胚珠cDNA文库中获得了13,505个EST，其中966个(7.15%)含有一个或多个SSR，从中开发出489对EST-SSR引物。在EST-SSR中，59.1%是三核苷酸，其次是二核苷酸(30%)、四核苷酸(6.4%)、五核苷酸(1.8%)和六核苷酸(2.7%)。AT/TA重复频率最高(18.4%)，其次为CTT/GAA(5.3%)、AG/TC(5.1%)、AGA/TCT(4.9%)、AGT/TCA(4.5%)和AAG/TTC(4.5%)。共扩增出130个EST-SSR基因座，在两个作图亲本之间产生了多态性。其中123个以杂交组合[(TM-1×海7124)TM-1]整合到我们的异源四倍体棉花遗传图谱上。EST-SSR标记分布在20条染色体和6个连锁群上。这些EST-SSR标记可用于棉花遗传作图、数量性状座位(QTL)定位和比较基因组学研究。
In order to construct a saturated genetic map and facilitate marker-assisted selection (MAS) breeding, it is necessary to enhance the current reservoir of known molecular markers in Gossypium. Microsatellites or simple sequence repeats (SSRs) occur in expressed sequence tags (EST) in plants (Kantety et al., Plant Mol Biol 48:501-510, 2002). Many ESTs are publicly available now and represent a good tool in developing EST-SSRs. From 13,505 ESTs developed from our two cotton fiber/ovule cDNA libraries constructed for Upland cotton, 966 (7.15%) contained one or more SSRs and from them, 489 EST-SSR primer pairs were developed. Among the EST-SSRs, 59.1% are trinucleotides, followed by dinucleotides (30%), tetranucleotides (6.4%), pentanucleotides (1.8%), and hexanucleotides (2.7%). AT/TA (18.4%) is the most frequent repeat, followed by CTT/GAA (5.3%), AG/TC (5.1%), AGA/TCT (4.9%), AGT/TCA (4.5%), and AAG/TTC (4.5%). One hundred and thirty EST-SSR loci were produced from 114 informative EST-SSR primer pairs, which generated polymorphism between our two mapping parents. Of these, 123 were integrated on our allotetraploid cotton genetic map, based on the cross [(TM-1xHai7124)TM-1]. EST-SSR markers were distributed over 20 chromosomes and 6 linkage groups in the map. These EST-SSR markers can be used in genetic mapping, identification of quantitative trait loci (QTLs), and comparative genomics studies of cotton.