梅菜ISSR分子标记技术初步研究

Preliminary Identification of Brassica Juncea Varieties by ISSR Molecular Markers

以梅菜为研究对象,初步建立起梅菜的ISSR分子标记技术体系,包括DNA提取、退火温度、循环次数和引物筛选等。利用CTAB法提取梅菜的DNA,经电泳检测和紫外检测,质量较好,适合PCR扩增。对PCR反应体系和条件进行优化,多态性引物分别为809,841,842,845,退火温度为58℃,次数为40次,Tag酶用量为0.8 U。本试验为梅菜的种子纯度鉴定和新品种选育提供技术依据。

The studies aimed to preliminary establish a technical system of ISSR molecular marker for Brassica Juneea varieties. It included the DNA extraction, optimization of PCR parameter such as the annealing temperature, cycle times, and primer screening etc. After being detected by electrophoresis and ultraviolet photometric,the CTAB is good to extract the DNA of Brassica Juncea and suitable for PCR. To optimize the PCR＇s reaction system and its conditions,we set the No. 809 ,No. 841 ,No. 842 and No. 845 as polymorphism primers, the annealing temperature is 58 22,40 cycles, the amount of Tag enzyme is 0.8 U. The testing will provide technical basis for seed purity of Brassica Juncea and new variety breeding