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稻属物种的染色体组类型有10种,其中具CD染色体组的物种有3种(Oryza alta Swallen,O.grandiglumis(Doell)Prod.和O.latifolia Desv.),仅分布在美洲中部和南部。这3个种在形态上和稻属其他染色体组的种比较容易区别,但它们彼此之间鉴别比较困难。近年来的研究表明,O.alta和O.grandiglumis应归并为1个种(O.grandiglumis),而O.latifolia应保持不变。本文基于代表不同分布区11个样品的核糖体转录间隔区(ITS)的77个克隆序列数据,利用DNA Strider1.2软件进行了限制性酶切位点分析,提出了一个鉴别CD染色体组物种的方法。方法的具体步骤是:(1)用通用引物扩增ITS片段;(2)用特异性的限制性内切酶Fok Ⅰ和/或Dra Ⅲ消化PCR扩增产物:(3)用1％的琼脂糖电泳并根据消化产物的多态性特征来鉴别不同物种。基于本文提出的核糖体ITS限制性片段多态性(RLFP)分析,可以快速和可靠地将稻属CD染色体组物种区别开来。
There are 10 genomic types of the genus Oryza, of which there are 3 species with the CD genome (Oryza alta Swallen, O. grandiglumis (Doell) Prod. And O. latifolia Desv.), Distributed only in the central and southern Americas. The three species are relatively easy to distinguish between morphologically and other genomes of the genus Oryza, but they are more difficult to distinguish from each other. Recent studies have shown that O. alta and O. grandiglumis should be integrated into one species (O. grandiglumis), while O. latifolia should remain unchanged. Based on the 77 cloned sequence data of ribosomal RNA transcribed spacer region (ITS) representing 11 samples in different distribution areas, we analyzed the restriction enzyme sites using DNA Strider1.2 software and proposed a method to identify the species of CD genome method. The specific steps of the method are: (1) amplifying ITS fragments with universal primers; (2) digesting PCR products with specific restriction enzymes Fok I and / or Dra III: (3) Sugar electrophoresis and to identify different species based on the polymorphism of digestive products. Based on the ribosome ITS restriction fragment polymorphism (RLFP) analysis presented in this paper, it is possible to quickly and reliably distinguish between the Oryza genus CD genomic species.