Development of RAPD-SCAR markers for Lonicera japonica (Caprifolicaceae) variety authentication by improved RAPD and DNA cloning
Development of RAPD-SCAR markers for Lonicera japonica (Caprifolicaceae) variety authentication by improved RAPD and DNA cloning
复制标题
通过改进的 RAPD 和 DNA 克隆开发用于忍冬品种鉴定的 RAPD-SCAR 标记。
DOI：
10.15517/rbt.v62i4.13493
复制
发表时间：
2014-12-01
期刊：
影响因子：
--
通讯作者：
Fu, Junjiang
中科院分区：
文献类型：
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作者：
Yang, Luquan;Khan, Md. Asaduzzaman;Fu, Junjiang
遗传多样性是一个物种的共同特征，它对物种的生存和适应起着至关重要的作用，对物种的识别和鉴定具有重要意义。日本野牡丹是一种传统的药用植物，近年来，利用改进的随机扩增多态性DNA（RAPD）技术对该植物进行了遗传学研究。本研究以这些RAPD片段为基础，建立了一套用于鉴定L.粳稻品种从不同样品的新鲜嫩叶中提取DNA。采自中国深圳、宜昌、乐山、峨眉、娄底等地。DNA材料采用改良的RAPD PCR扩增。通过对不同物种的RAPD扩增片段进行切取、克隆和扩增，建立了适合不同物种的稳定的序列特征性扩增区域（SCAR）标记。从改良的RAPD片段中成功克隆了两个SCAR标记JYH 3 -3和JYH 4 -3。SCAR标记JYH 3 -3对所有的L. JYH 4-3是来自广东省深圳市的一个特异标记，深圳市与湖北、四川、湖南三省相距甚远，是其它L. japonica samples）。筛选出JYH 3 -3标记作为鉴定该菌的特异分子标记。JYH 4 -3是深圳地区的分子标记。所开发的SCAR标记可作为苜蓿的特异性分子标记。粳稻品种鉴定改进的RAPD分析与SCAR标记的结合为研究生物的遗传多样性提供了有力的工具，我们在L.日本的。
Genetic diversity within a species is a common feature, which plays a vital role in its survival and adaptability, and is important for the identification and authentication of a species. Lonicera japonica is a traditionally used medicinal plant, which have been recently genetically characterized by an improved ran- dom amplified polymorphic DNA (RAPD) analysis. In this study, the molecular markers on the basis of these RAPD fragments have been developed to identify specific L. japonica variety. The DNAs were extracted from fresh young leaves of different samples of L. japonica collected from Shenzhen, Yichang, Leshan, Emei and Loudi, China. The DNA materials were amplified using improved RAPD PCR. Different RAPD bands were excised, cloned and developed for stable sequence-characterized amplified region (SCAR) markers with differ- ent species. Two SCAR markers, JYH3-3 and JYH4-3, have been successfully cloned from improved.RAPD fragments. The SCAR marker JYH3-3 was found specific for all of the L. japonica samples collected from the different regions, and another marker JYH 4-3 was strictly specific to the Shenzhen sample from Guangdong province, which is geographically distant from Hubei, Sichuan and Hunan Provinces (source of other L. japonica samples). The marker JYH3-3 was found as specific molecular marker for the identification of L. japonica, while JYH4-3 was found as molecular marker strictly specific for the Shenzhen sample. The developed SCAR markers might serve as more specific molecular markers for L. japonica variety authentication. The combination of improved RAPD analysis and SCAR marker development have resulted useful tools to study the genetic variety of any organism, which we have successfully applied here in L. japonica.