维吾尔药茶昆仑雪菊ISSR分子标记体系的建立

研究适用于昆仑雪菊的ISSR-PCR反应体系,建立ISSR分子标记技术评价昆仑雪菊生态多样性体系。采用改良CTAB法、CTAB区室法、改良CTAB区室法、试剂盒法提取昆仑雪菊的新鲜叶片、干花以及种子的DNA;以827为引物,研究浓度、循环数、退火温度对ISSR-PCR反应体系的影响,以不同实验材料检测其适宜性。结果表明,昆仑雪菊新鲜叶片、干花以及种子的DNA的最佳提取方法为改良CTAB区室法,ISSR-PCR反应体系的最适引物浓度为1.0μmol·L-1,退火温度为54.6℃,循环数为30。ISSR-PCR反应体系适用于评价昆仑雪菊的生物多样性。

An ISSR Molecular Labeling Technique System for Uygur Herbal Tea Kunlun Chrysanthemum

The research aimed to study the ISSR-PCR reaction system and establish the ISSR molecular marker technique to evaluate the ecological diversity of Kunlun chrysanthemum.Genomic DNA was extracted from the fresh leaves,dried flowers and seeds of Kunlun chrysanthemum by four different DNA extraction methods as following:improved CTAB method,CTAB subarea method,improved CTAB subarea method and kit method.Primer 827 was used to optimize concentration,cycle time,annealing temperature on the ISSR-PCR reaction system,and the suitability of the methods was tested by different materials.The best extraction method of fresh leaves,dried flowers and seeds was the improved CTAB subarea method,the optimal primer concentration was 1.0 μmol · L1,the best annealing tem perature was 54.6 ℃C,and the perfect cycle time was 30.The ISSR-PCR reaction system is suitable for evaluating the biodiversity of Kunlun chrysanthemum.