Identifying and distinguishing sibling species in the Tetrahymena pyriformis complex (Ciliophora, Oligohymenophorea) using PCR/RFLP analysis of nuclear ribosomal DNA
Identifying and distinguishing sibling species in the Tetrahymena pyriformis complex (Ciliophora, Oligohymenophorea) using PCR/RFLP analysis of nuclear ribosomal DNA
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DOI：
10.1111/j.1550-7408.1996.tb04509.x
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发表时间：
1996-11-01
影响因子：
2.2
通讯作者：
Lynn, DH
中科院分区：
文献类型：
--
作者：
Jerome, CA;Lynn, DH
我们描述了一个riboprinting策略，用于识别和区分兄弟物种的四膜虫梨形复合体。它涉及使用聚合酶链反应扩增大部分核核糖体DNA和内部转录间隔区，并用限制性内切酶消化该DNA。然后使用独特的限制性片段长度模式或单倍型将物种区分为：（1）在物种水平上可鉴定的六个分类群，（2）分成四对的八个分类群，和（3）彼此相同的一组八个分类群。后一个结果表明，需要一个更可变的分子来区分复杂的最密切相关的物种。有两个菌株从一个物种（嗜热四膜虫），也没有从另一个物种（四膜虫empidokyrea）的多个菌株之间的种内变异。这种方法提供了一种替代传统的技术来识别T。梨形种需要活的参考标本和/或揭示高水平的种内变异。
We describe a riboprinting strategy for identifying and distinguishing among sibling species in the Tetrahymena pyriformis complex. It involves use of the polymerase chain reaction to amplify a large segment of the nuclear ribosomal DNA and internal transcribed spacers, and digestion of this DNA with restriction enzymes. Unique restriction fragment length patterns or haplotypes were then used to distinguish species into: (1) six taxa that were identifiable to the species level, (2) eight taxa that were separated into four pairs, and (3) a group of eight taxa that were identical to each other. The latter result indicates that a more variable molecule is needed to distinguish the most closely related species in the complex. There was no intraspecific variation between two strains from one species (Tetrahymena thermophila) nor among multiple isolates from another species (Tetrahymena empidokyrea). This approach provides an alternative to traditional techniques for identifying T. pyriformis species that require living reference specimens and/or that reveal high levels of intraspecific variation.