We reported our preliminary study on mapping of the expressed sequence tag (EST) using single-stranded DNA conformation polymorphism (SSCP) on polymerase chain reaction (PCR) product. Five ESTs primer pairs, developed...
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We reported our preliminary study on mapping of the expressed sequence tag (EST) using single-stranded DNA conformation polymorphism (SSCP) on polymerase chain reaction (PCR) product. Five ESTs primer pairs, developed from nuclear genes including CAD(cinnamyl-alcohol dehydrgenase), CHS(chalcone synthase), NIR(nitrite reductase), ARA554 (cDNA expressed in differentiating xylem in Pinus taedae) and GS(glutamine synthetase), were selected to optimize the experimental protocol and generate mapping markers in the megagemtophytes from a single tree of Pinus massoniana. Our ultimate goal was to use SSCP approach to construct a transcriptional map for comparative mapping studies in P. massoniana. The efficiency to construct a transcriptional map in P. massoniana based on the published EST primer pairs derived from other pine species critically depended on the successful amplification of EST fragments and the ratio of the heterozygous loci revealed in ***. In this study, 3 (60) out of 5 tested EST primer pairs were succeeded in amplification, however, only 1(20) gene was heterozygous in the tested tree. The ratio of heterozygous loci detected in this study was similar to that revealed by anonymous marker in ***. Therefore, a low efficiency would be expected if the map would be constructed using single pedigree. The segregation ratio of loci revealed by primer pairs would be higher if multiple pedigrees would be used. We proposed that consensus mapping approach based on multiple-pedigree should be used for EST mapping and therefore to increase the efficiency of EST mapping.
利用PCR扩增产物直接测序的方法分析广义青篱竹属 (Arundinaria)中有关争议类群的代表种或模式种 (毛竹为外类群 )等 18种竹种的核糖体DNA内转录间隔区 (InternalTranscribedSpacers ,ITS)序列。通过最简约性分析产生的ITS系统发育树表明 ,供试竹种形成一个自然的单系类群 ,这说明广义青篱竹属中这些不同的类群归属青篱竹属是合理的。 17种竹种可聚为 2大分支 :其中斑苦竹 (A .oleosa)、仙居苦竹 (A .hsienchuensis)、茶秆竹 (A .amabilis)、长叶苦竹 (A .chino)、苦竹 (A .amara)、宜兴苦竹 (A .yixingensis)、菲白竹 (A .fortunei)、翠竹 (A .pygmaea)为一个分支 ;而大明竹 (A .graminea)、巴山木竹 (A .fargesii)、冷箭竹 (A .faberi)、凤竹 (A .hupehense)、鼓节矢竹 (***)、矢竹 (Pseudosasajaponica)、短穗竹 (Brachystachyumdensiflorum)、肿节竹 (A .oedogonata)、少穗竹 (A .sulcata)组合在另一分支。ITS系统发育树还表明 ,大明竹与巴山木竹、鼓节矢竹与矢竹、少穗竹与短穗竹和肿节竹关系极为密切 ,均得到较高的Bootstrap(分别为 99%、10 0 %和 87% )的支持 ;茶秆竹与仙居苦竹关系非常密切 ,茶秆竹可归隶到青篱竹属中 ;翠竹和菲白竹关系密切 。
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