Species identification in seven small millet species using polymerase chain reaction - Restriction fragment length polymorphism of trnS-psbC gene region
Species identification in seven small millet species using polymerase chain reaction - Restriction fragment length polymorphism of trnS-psbC gene region
复制标题
聚合酶链式反应对七种小谷子进行物种鉴定——trnS-psbC基因区限制性片段长度多态性
作者：
M. Parani;K. Rajesh;M. Lakshmi;L. Parducci;A. Szmidt;A. Parida
采用聚合酶链反应（PCR）技术，对来自7个小型谷子品种的119份材料的叶绿体trnS-psbC基因进行扩增，并用8种限制性内切酶分别或组合进行限制性片段长度多态性（PCR-RFLP）分析。PCR-RFLP与个别酶之间的多态性，只有一些物种。然而，所有的物种都可以通过使用两种酶的组合来区分，特别是HaeIII和MspI。对11~20份具有相同酶组合的材料进行PCR-RFLP分析，结果表明，种内无变异，说明差异带型具有种间特异性。与此相反，相同的酶组合是不是有用的区分不同种的木豆属，Rhyncosia，冷杉属，Rhizophora，Ceriops，和木榄属，它也揭示了种内变异在3种冷杉。本研究表明，用两种四碱基识别酶消化trnS-psbC比单独使用任何一种酶显示更多的变化，这可能是一种方法，在一些属的物种鉴定的选择。
The chloroplast trnS-psbC gene regions from total genomic DNA of 119 accessions from seven small millet species were amplified by polymerase chain reaction (PCR) and digested with eight restriction enzymes individually as well as in combinations of two enzymes to generate restriction fragment length polymorphism (PCR-RFLP). PCR-RFLP with individual enzymes revealed polymorphism between only some species. However, all the species could be distinguished by using a combination of two enzymes, specifically HaeIII and MspI. PCR-RFLP of 11 to 20 accessions with the same enzyme combination showed no intraspecific variation, which established that the differential banding patterns were species specific. In contrast, the same enzyme combination was not useful for differentiating different species of the genera Cajanus, Rhyncosia, Abies, Rhizophora, Ceriops, and Bruguiera, and it also revealed intraspecies variation in three species of Abies. The present study indicated that digestion of trnS-psbC with two four-base recognizing enzymes reveals more variation than with either enzyme alone and that it may be a method of choice for species identification in some genera.