Molecular divergence and application of the ITS-5.8S rDNA and RUBISCO spacer in Porphyra haitanensis Chang et Zheng (Bangiales, Rhodophyta)
Molecular divergence and application of the ITS-5.8S rDNA and RUBISCO spacer in Porphyra haitanensis Chang et Zheng (Bangiales, Rhodophyta)
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DOI：
10.1007/s10499-010-9322-y
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发表时间：
2010-01
影响因子：
2.9
通讯作者：
Chang-sheng Chen;Chaotian Xie;De-hua Ji;Yan Liang;Lingmin Zhao
中科院分区：
文献类型：
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作者：
Chang-sheng Chen;Chaotian Xie;De-hua Ji;Yan Liang;Lingmin Zhao
为了选择一种可靠、灵敏的海地紫菜菌株鉴别方法，比较了5株野生和5株栽培紫菜菌株核糖体DNA内转录间隔区1至内转录间隔区（ITS-5.8S）核苷酸序列和RUBISCO基因间间隔区。基于分子分析，ITS-5.8S序列（约1210 bp）可划分为ITS1、5.8S和ITS2三个区域。每个菌株的ITS1和ITS2序列不同，甚至在同一地点采集的个体之间也是如此。相比之下，10株海地紫菜菌株的5.8S rDNA和RUBISCO间隔序列是相同的，但不同种类紫菜之间存在差异。系统发育分析也支持这些结论。这些在ITS-5.8S序列中重复出现的高度保守区和多样化区序列特征，可用于海地紫菜品系或紫菜种的种质鉴别。相比之下，RUBISCO间隔只适用于鉴别卟啉种类。设计了新的偶联引物，只扩增了卟啉的5.8S rDNA和ITS2区域。这些扩增片段的序列可以很容易地用于种质鉴定或进行卟啉的系统发育分析。
To select a reliable and sensitive method for discriminating strains of Porphyra haitanensis, the nucleotide sequence of the internal transcribed spacer 1 to internal transcribed spacer 2 regions (ITS-5.8S) of nuclear ribosomal DNA and the intergenic spacer region of RUBISCO were compared in five wild and five cultivated Porphyra haitanensis strains. Based on molecular analyses, sequences of ITS-5.8S (about 1,210 bp) could be divided into three regions: ITS1, 5.8S, and ITS2. The ITS1 and ITS2 sequences of each strain differed, even between individuals collected from the same site. In contrast, 5.8S rDNA and RUBISCO spacer sequences were identical among the ten P. haitanensis strains, although differences were found among different Porphyra species. Phylogenetic analysis also supported these conclusions. These sequence features of highly conserved regions and diversified regions that occurred repeatedly in ITS-5.8S could be useful in discriminating germplasm of P. haitanensis strains or Porphyra species. In contrast, the RUBISCO spacer is only suitable for identifying Porphyra species. New coupled primers were designed to amplify only the 5.8S rDNA and ITS2 region of Porphyra. The sequences of these amplified fragments can be readily used to identify germplasm or to perform phylogenetic analysis of Porphyra spp.