Pastelín Solano Miriam Cristina, Salinas Ruíz Josafhat, González Arnao María Teresa, Castañeda Castro Odón, Galindo Tovar María Elena, Bello Bello Jericó Jabín
1Faculty of Biological and Agropecuary Sciences, University of Veracruz, Km 1 Peñuela-Amatlán de los Reyes, Federal Highway, 94950 Amatlán de los Reyes, State of Veracruz Mexico.
Postgraduate College-Campus Córdoba, Km 348 Córdoba-Veracruz Federal Highway, 94946 Amatlán de los Reyes, State of Veracruz Mexico.
Physiol Mol Biol Plants. 2019 Mar;25(2):561-567. doi: 10.1007/s12298-019-00645-9. Epub 2019 Feb 2.
The effect of subculture cycles on somaclonal variation of using intersimple sequence repeat (ISSR) markers was analyzed. Nodal segments of 2 cm in length were established in vitro and multiplied by 10 subculture cycles in Murashige and Skoog (MS) medium supplemented with 8.86 μM BAP (benzylaminopurine). After 45 days in each culture, the length and number of shoots per explant were evaluated. For ISSR markers, ten shoots per each subculture and the mother plant were used. Ten ISSR primers were used and a total of 118 bands were obtained. The polymorphism (%) was calculated and a dendrogram based on Jaccard's genetic distance between the subcultures and the donor plant was obtained. These results show that the multiplication rate tends to increase until subculture five, whereas shoot length decreases as the number of subcultures increases. The ISSR markers revealed an increase in the polymorphism percentage after the fifth culture cycle. The dendrogram showed the formation of two groups. The first group, with less genetic variability, is the donor plant and subcultures 1-5; the second group has greater genetic distance and is formed by subcultures 6-10. The results revealed that the number of subcultures with 8.86 μM BAP is a factor that affects the somaclonal variation during in vitro regeneration of . In conclusion, the subculture number affects somaclonal variation and in vitro development of .
分析了继代培养周期对使用简单序列重复区间（ISSR）标记的体细胞克隆变异的影响。将2厘米长的茎段在体外培养，并在添加了8.86μM苄氨基嘌呤（BAP）的Murashige和Skoog（MS）培养基中进行10次继代培养繁殖。每次培养45天后，评估每个外植体的芽长和芽数。对于ISSR标记，每次继代培养选取10个芽以及母株进行分析。使用了10条ISSR引物，共获得118条带。计算多态性（%），并基于继代培养物与供体植物之间的杰卡德遗传距离得到一个聚类图。这些结果表明，增殖率在继代培养5次之前趋于增加，而随着继代培养次数的增加，芽长会降低。ISSR标记显示在第5次培养周期后多态性百分比增加。聚类图显示形成了两组。第一组遗传变异性较小，包括供体植物和第1 - 5次继代培养物；第二组遗传距离较大，由第6 - 10次继代培养物组成。结果表明，添加8.86μM BAP的继代培养次数是影响[植物名称未给出]体外再生过程中体细胞克隆变异的一个因素。总之，继代培养次数影响[植物名称未给出]的体细胞克隆变异和体外发育。
