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基于核糖体内部转录间隔区(ITS)测序的药用栽培品种杂种特性研究。

Study on Hybrid Characteristics of Medicinally Used Cultivated Species Using Ribosomal Internal Transcribed Spacer (ITS) Sequencing.

机构信息

Institute of Soil and Water Conservation, CAS & MWR, Yangling 712100, China.

College of Landscape Architecture, Zhejiang Agriculture & Forestry University, Hangzhou 311300, China.

出版信息

Molecules. 2018 Jun 28;23(7):1565. doi: 10.3390/molecules23071565.

Abstract

taxa, as a traditional Chinese medicinal and edible plant, has found expanding domestic and foreign applications in recent decades. However, the poor management in germplasm resources market has inevitably caused an unnecessary hybrid of the provenances. In order to clarify the hybrid characteristics of germplasm resources in the main production area, the cultivars collected from the provinces Gansu, Shannxi, Shanxi, and Hubei of China were researched, using internal transcribed spacer (ITS) sequence technology. The confirmation of additive nucleotides based on the ITS sequencing of polymerase chain reaction (PCR) mixture was optimized and used to study the hybrid of cultivars. The results showed that when the ratio of PCR mixture increased up to 15 percent, the presence of a double peak in the sequencing electrophoresis map could be confirmed, suggesting the existence of additive nucleotides. According to the method above, 46 samples of cultivars collected during 2016 and 2017 were studied and compared with the samples collected from the year 2009 to 2010. All of the samples collected during 2016 and 2017 were hybridized and no genetic pure lines were found. In addition, the sites of variable base reduced greatly, concentrating at positions 122 and/or 226. These phenomena suggested that the genetic diversity of cultivars declined and the germplasm resources gradually converged. More attention should be paid to the reasonable exploitation and genetic breeding of taxa.

摘要

作为一种传统的中药材和食用植物,菝葜在近几十年来在国内外得到了广泛的应用。然而,由于种源资源市场管理不善,不可避免地导致了种源的不必要杂交。为了澄清主要产区种质资源的杂种特征,本研究采用内转录间隔区(ITS)序列技术,对来自中国甘肃、陕西、山西和湖北四省的栽培品种进行了研究。基于聚合酶链反应(PCR)混合物 ITS 测序的加性核苷酸的确认进行了优化,并用于研究栽培品种的杂种情况。结果表明,当 PCR 混合物的比例增加到 15%时,可以确认测序电泳图谱中存在双峰,表明存在加性核苷酸。根据上述方法,对 2016 年和 2017 年收集的 46 个菝葜品种样本进行了研究,并与 2009 年至 2010 年收集的样本进行了比较。所有 2016 年和 2017 年收集的样本都发生了杂交,没有发现遗传纯系。此外,可变碱基的位置大大减少,集中在 122 位和/或 226 位。这些现象表明,菝葜品种的遗传多样性下降,种质资源逐渐趋同。应该更加关注菝葜的合理开发和遗传育种。

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