Patel Hardik K, Fougat Ranbir S, Kumar Sushil, Mistry Jigar G, Kumar Mukesh
Department of Agricultural Biotechnology, Anand Agricultural University, Anand, 388 110, India.
3 Biotech. 2015 Oct;5(5):697-707. doi: 10.1007/s13205-014-0269-y. Epub 2014 Dec 9.
There is a lack of information on the molecular characterization of Ocimum species and hence, efforts have been made under the present study to characterize 17 Ocimum genotypes belonging to 5 different species (O. basilicum, O. americanum, O. sanctum, O. gratissimum and O. Polystachyon) through random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) markers. PCR amplification using 20 RAPD primers generated a total of 506 loci, of which 490 (96.47 %) loci were found polymorphic. The PIC value for RAPD ranged from 0.907 (OPF 14) to 0.954 (OPC 11) with an average of 0.937. The ISSR primers generated a total of 238 loci, of them 234 (98.17 %) loci were polymorphic. The PIC value ranged from 0.892 (UBC 808) to 0.943 (ISSR A12) with an average of 0.923. The average Jaccard's similarity coefficient based on RAPD and ISSR analysis was 0.58 and 0.52, respectively. Clustering pattern of dendrogram generated using the pooled RAPD and ISSR data showed all Ocimum genotypes in their respective species groups at a cutoff value of 0.49 and 0.42, respectively. Many unique species-specific alleles were amplified by RAPD and ISSR markers. In both marker systems, a maximum number of unique alleles were observed in O. sanctum. The results of the present investigation provided valid guidelines for collection, conservation and characterization of Ocimum genetic resources.
关于罗勒属植物的分子特征缺乏相关信息,因此,在本研究中,我们通过随机扩增多态性DNA(RAPD)和简单重复序列区间(ISSR)标记,对属于5个不同物种(罗勒、美国罗勒、圣罗勒、丁香罗勒和多穗罗勒)的17个罗勒基因型进行了特征分析。使用20条RAPD引物进行PCR扩增,共产生了506个位点,其中490个(96.47%)位点具有多态性。RAPD的PIC值范围为0.907(OPF 14)至0.954(OPC 11),平均值为0.937。ISSR引物共产生了238个位点,其中234个(98.17%)位点具有多态性。PIC值范围为0.892(UBC 808)至0.943(ISSR A12),平均值为0.923。基于RAPD和ISSR分析的平均杰卡德相似系数分别为0.58和0.52。使用合并的RAPD和ISSR数据生成的聚类图的聚类模式显示,所有罗勒基因型分别在各自的物种组中,截断值分别为0.49和0.42。RAPD和ISSR标记扩增出了许多独特的物种特异性等位基因。在这两种标记系统中,圣罗勒中观察到的独特等位基因数量最多。本研究结果为罗勒遗传资源的收集、保存和特征分析提供了有效的指导方针。