Development of RAPD-SCAR markers for different Ganoderma species authentication by improved RAPD amplification and molecular cloning
Development of RAPD-SCAR markers for different Ganoderma species authentication by improved RAPD amplification and molecular cloning
复制标题
通过改进的 RAPD 扩增和分子克隆，开发用于不同灵芝物种鉴定的 RAPD-SCAR 标记。
DOI：
10.4238/2015.may.25.19
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发表时间：
2015-01-01
影响因子：
0.4
通讯作者：
Cheng, J. L.
中科院分区：
文献类型：
--
作者：
Fu, J. J.;Khan, M. A.;Cheng, J. L.
序列特征扩增区（SCAR）技术是一种很有价值的分子生物学技术。该方法主要来源于从随机扩增多态性DNA（RAPD）获得的扩增DNA片段的分子克隆。本研究以10种灵芝菌为材料，采用改良的RAPD技术对它们的DNA进行扩增。然后将扩增的片段克隆到T载体中，筛选阳性克隆，鉴定并测序用于SCAR标记的开发。通过设计PCR引物和优化PCR条件，开发了4个对赤灵芝（LZ 1 - 4和LZ 8 - 2）、华芝（LZ 2 - 2和LZ 8 - 2）、热带赤芝（LZ 8 - 2）和灵芝HG（LZ 9 - 15）具有特异性的SCAR标记，分别命名为LZ 1 - 4、LZ 2 - 2、LZ 8 - 2和LZ 9 - 15。这4个新的SCAR标记已在GenBank中登录，登录号为：分别为KM 391935、KM 391936、KM 391937和KM 391938。因此，在本研究中，我们开发了特定的SCAR标记，用于不同灵芝物种的鉴定和认证。
The sequence-characterized amplified region (SCAR) is a valuable molecular technique for the genetic identification of any species. This method is mainly derived from the molecular cloning of the amplified DNA fragments achieved from the random amplified polymorphic DNA (RAPD). In this study, we collected DNA from 10 species of Ganoderma mushroom and amplified the DNA using an improved RAPD technique. The amplified fragments were then cloned into a T-vector, and positive clones were screened, indentified, and sequenced for the development of SCAR markers. After designing PCR primers and optimizing PCR conditions, 4 SCAR markers, named LZ1-4, LZ2-2, LZ8-2, and LZ9-15, were developed, which were specific to Ganoderma gibbosum (LZ1-4 and LZ8-2), Ganoderma sinense (LZ2-2 and LZ8-2), Ganoderma tropicum (LZ8-2), and Ganoderma lucidum HG (LZ9-15). These 4 novel SCAR markers were deposited into GenBank with the accession Nos. KM391935, KM391936, KM391937, and KM391938, respectively. Thus, in this study we developed specific SCAR markers for the identification and authentication of different Ganoderma species.