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利用简单重复序列间分子标记(ISSR)方法对铁皮石斛、叠鞘石斛、金钗石斛3种石斛种间、同种石斛不同蒴果间、同一石斛蒴果不同植株间及同种石斛不同继代次数组培苗的遗传多样性进行分析.结果表明:(1)3种石斛组培苗种间多态性位点百分率为80.59%,相似系数为0.32,在物种水平上遗传多态性处较高水平.铁皮石斛与金钗石斛的遗传距离最小,为0.406;铁皮石斛与叠鞘石斛的遗传近距离次之,为0.965;金钗石斛与叠鞘石斛的遗传距离最大,为1.099.(2)在同种石斛不同蒴果间,金钗石斛组培苗间遗传差异性最大,多态性位点百分率为77.78%,铁皮石斛次之,为58.82%,叠鞘石斛最低,为28.57%.造成这一现象的原因可能是金钗石斛蒴果数量最多,铁皮石斛次之,叠鞘石斛数量最少.相应地,不同蒴果间的遗传距离分别为0.338、0.301、0.191.(3)同种石斛同一蒴果不同植株间亦存在遗传差异性,金钗石斛不同植株间多态性位点百分率为最大,为25.00%,叠鞘石斛次之,为11.11%,铁皮石斛最小,为7.14%.它们之间的遗传距离分别为0.207、0.047、0.030.(4)3种石斛组培苗在4次继代培养过程中电泳条带带型一致,并未发生质的变化,认为没有发生遗传变异.可见,ISSR分子标记方法可较好地应用于石斛组培苗遗传差异性研究.
Using the simple sequence repeat marker (ISSR) method, the succession times of different generations of Dendrobium candidum, Dendrobium candidum, Dendrobium nobile Dendrobium species, the same capsule of Dendrobium candidum, different plants of the same Dendrobium capsule and the same Dendrobium species The results showed that: (1) The percentage of polymorphic loci in three Dendrobium species was 80.59% and the similarity coefficient was 0.32, indicating that the genetic polymorphism at the species level was higher at the genetic polymorphism The genetic distance between Dendrobium candidum and Dendrobium nobile was the smallest, which was 0.406, the genetic distance between Dendrobium candidum and Dendrobium candidum was 0.965, and the genetic distance between Dendrobium nobile and Dendrobium candidum was the largest, which was 1.099. The same species of Dendrobium between the capsule, Dendrobium nobile tissue culture seedling the genetic differences between the largest, the percentage of polymorphic loci was 77.78%, followed by Dendrobium candidum, 58.82%, Dendrobium lamella lowest, accounting for 28.57%. The reason of the phenomenon may be that the number of Dendrobium nobile is the largest, followed by Dendrobium candidum and Dendrobium candidum the least, correspondingly, the genetic distances among different capsules are 0.338,0.301,0.191, respectively. (3) The same capsule with the same species of Dendrobium There are also genetic differences between, nobile The percentage of polymorphic loci in different plants was 25.00%, followed by Dendrobium, 11.11%, and Dendrobium candidum the smallest, which was 7.14% .The genetic distances among them were 0.207, 0.047 and 0.030 respectively. 4) The three bands of Dendrobium candidum were the same in the fourth subculturing process, and there was no change in the quality of the three kinds of Dendrobium candidum, which means no genetic variation occurred.It can be seen that the ISSR molecular marker method can be applied to Dendrobium Study on Genetic Difference of Plantlets.