Santos M D M, Buso G C S, Torres A C
Departamento de Botânica, Universidade de Brasília, Brasília, DF, Brasil.
Genet Mol Res. 2008 Oct 21;7(4):1097-105. doi: 10.4238/vol7-4gmr489.
The objective of the present study was to evaluate the genetic variability in micropropagated plantlets of ornamental pineapple, after the fourth period of subculture. The basal culture medium consisted of MS salts, vitamins, 3% sucrose, liquid formulation, supplemented with 6-benzylaminopurine (BAP) at concentrations of 0.125, 0.25, 0.5, 1.0, and 2.0 mg/L. The addition of BAP influenced the occurrence of genetic variation revealed using random amplified polymorphic DNA (RAPD) markers. Of a total of 520 primers tested, 44 were selected and amplified; 402 monomorphic bands (97.2%) and 18 polymorphic bands (2.8%) resulted among regenerated plantlets. The polymorphic fragments were produced by 12 primers (OPA-01, OPA-20, OPB-01, OPB-19, OPC-19, OPF-13, OPL-17, OPM-13, OPP-16, OPT-07, OPV-19, and OPX-03). Among the primers that identified polymorphism, OPA-01, OPA-20, OPB-19, OPC-19, OPL-17, OPP-16, and OPX-3 each showed, one polymorphic band and OPF-13 amplified a maximum of three bands. In this study, the RAPD technique was effective in showing the occurrence of somaclonal variations that occur during the micropropagation process of ornamental pineapple cultivation in BAP-supplemented medium, and it is possible to detect the presence of genetic variation in early stages of plant development.
本研究的目的是评估观赏菠萝微繁殖苗在继代培养第四个周期后的遗传变异性。基本培养基由MS盐、维生素、3%蔗糖、液体配方组成,并添加浓度为0.125、0.25、0.5、1.0和2.0 mg/L的6-苄基腺嘌呤(BAP)。BAP的添加影响了利用随机扩增多态性DNA(RAPD)标记揭示的遗传变异的发生。在总共测试的520个引物中,选择并扩增了44个;再生苗中产生了402条单态带(97.2%)和18条多态带(2.8%)。多态性片段由12个引物(OPA-01、OPA-20、OPB-01、OPB-19、OPC-19、OPF-13、OPL-17、OPM-13、OPP-16、OPT-07、OPV-19和OPX-03)产生。在鉴定出多态性的引物中,OPA-01、OPA-20、OPB-19、OPC-19、OPL-17、OPP-16和OPX-3各自显示一条多态带,OPF-13最多扩增出三条带。在本研究中,RAPD技术有效地显示了在添加BAP的培养基中观赏菠萝栽培微繁殖过程中发生的体细胞克隆变异的发生情况,并且有可能在植物发育的早期阶段检测到遗传变异的存在。