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以桉树的赤桉、巨桉、细叶桉和粗皮桉4个种18个种源为研究材料,根据优化的ISSR-PCR反应体系和扩增条件,从100条引物中筛选出12条扩增条带特异、条带数适中、背景清晰、重复性好的引物,总计扩增出120条清晰可重复的DNA条带,其中93条是多态性条带。每个引物可扩增的多态条带长度范围为300~2 500 bp,数目在4~11条,平均为7.75条,多态带百分率为77.5%。通过软件POPGENE 1.31、MVSP 32软件和NTSYS-pc软件对所得数据进行分析,从而得出4个种18个种源的遗传相似系数,并绘制分子聚类图和主坐标分析图,结果表明,遗传一致度的变化范围在0.387 1~0.914 0之间,遗传距离的变化范围在0.089 9~0.949 1之间,种间遗传相似性差异较大,种内遗传相似性极高。
According to the optimized ISSR-PCR reaction system and amplification conditions, 12 provenances of eucalyptus, eucalyptus grandis, eucalyptus urophylla and eucalyptus urophylla were selected as the research materials, 12 primers were screened out from 100 primers A total of 120 clear and repeatable DNA bands were amplified using primers with specific bands, moderate number of bands, clear background and good repeatability. Among them, 93 were polymorphic bands. The length of polymorphic bands amplified by each primer ranged from 300 to 2500 bp, the number of which ranged from 4 to 11 with an average of 7.75 and the percentage of polymorphic bands was 77.5%. The data obtained from POPGENE 1.31 software, MVSP 32 software and NTSYS-pc software were analyzed to obtain the genetic similarity coefficients of 4 species and 18 provenances, and the molecular clustering and principal coordinates analysis were drawn. The results showed that genetic The range of variation was between 0.387 1 and 0.914 0, and the range of genetic distance was between 0.089 9 and 0.949 1. The genetic similarities among species varied greatly, and the intraspecies genetic similarity was very high.