PCR-RFLP鉴别假高粱和丝克高粱

本文利用PCR特异引物扩增rDNA-18S-26S基因,对PCR产物进行纯化、克隆和测序;分析假高粱与丝克高粱rDNA-18S-26S的酶切图谱,选择合适的限制性内切酶对两者PCR产物进行特异性切割,按照酶切片段的长度来区分两个近似种.被AflIII酶切后,假高粱被切割为674bp、174bp两个片段,而丝克高粱被切割成181bp、174bp、495bp、669bp 4个片段.为建立PCR-RFLP分子鉴定方法有效地区分假高粱与丝克高粱这两个种提供依据.

Differentiation of Sorghum halepense and S.silk by PCR-RFLP method

To establish a molecular method for differentiation of Sorghum halepense and S. silk, poly-merase chain reaction-restriction fragment length polymorphism analysis was used. The rDNA-18S-26S gene was amplified by specific primers and PCR products were purified, cloned , sequenced and analyzed. Afl III restriction enzyme was chosen based on the restriction maps of rDNA-18S-26S gene of S. halepense and S. silk to cut PCR products. Results showed that two different rDNA-18S-26S sequences blasted as S . halepense and S . silk were detected in the study. S . halepense and S . silk can be visuallly distinguished according to the electrophoresis pattern: the four fragments in 181bp, 174bp, 495bp and 669bp were observed in the lanes of S. silk while only two bands in 674bp and 174bp for S. halepense . Therefore, this PCR-RFLP method established in this study can be used as the basis for identification between S. halepense and S. silk .