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苎麻长白毛变种和苎麻短白毛变种的分子标记的开发与验证。

Development and validation of molecular markers for characterization of Boehmeria nivea var. nivea and Boehmeria nivea var. tenacissima.

机构信息

Department of Life Sciences, Tzu-Chi University, Hualien 970, Taiwan.

出版信息

Chin Med. 2010 Nov 29;5:40. doi: 10.1186/1749-8546-5-40.

DOI:10.1186/1749-8546-5-40
PMID:21114822
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3004904/
Abstract

BACKGROUND

The root of Boehmeria spp (ramie) is a hepatoprotective Chinese herbal medicine. Medicinal properties vary between Boehmeria nivea var. nivea and Boehmeria nivea var. tenacissima, which are local species found in Taiwan. As commercial preparations may use either species, there is a need for a rapid and simple assay to identify variants for quality control.

METHODS

Four methods were developed and tested for their applicability in differentiating the two species. These methods were random amplified polymorphic DNA (RAPD); sequence characterized amplified regions (SCAR); single nucleotide polymorphisms (SNP) and cleaved amplified polymorphic sequences (CAPS).

RESULTS

Three RAPD markers were developed that produced unique bands in B. nivea var. tenacissima and B. nivea var. nivea. Based on sequenced RAPD bands, one SCAR marker was developed that produced a single DNA band in B. nivea var. nivea. Two SNP markers differentiated between B. nivea var. nivea and B. nivea var. tenacissima based on single nucleotide substitutions. A pair of CAPS oligonucleotides was developed by amplifying a 0.55-kb DNA fragment that exhibited species-specific digestion patterns with restriction enzymes Alf III and Nde I. Consistent results were obtained with all the four markers on all tested Boehmeria lines.

CONCLUSION

The present study demonstrates the use of the RAPD, SCAR, SNP and CAPS markers for rapid identification of two closely related Boehmeria species.

摘要

背景

苎麻根是一种具有保肝作用的中草药。中国台湾产的苎麻有两种，即白叶苎麻和青麻，药用特性不同。由于商业制剂可能使用这两个种，因此需要一种快速简单的方法来识别变种，以进行质量控制。

方法

为了区分这两个种，开发并测试了 4 种方法。这些方法是随机扩增多态性 DNA（RAPD）；序列特征扩增区（SCAR）；单核苷酸多态性（SNP）和切割扩增多态性序列（CAPS）。

结果

开发了 3 个 RAPD 标记，它们在青麻和白叶苎麻中产生独特的条带。基于测序的 RAPD 条带，开发了一个 SCAR 标记，它在白叶苎麻中产生一个单一的 DNA 条带。基于单个核苷酸取代，两个 SNP 标记可以区分白叶苎麻和青麻。通过扩增 0.55kb 的 DNA 片段，设计了一对 CAPS 寡核苷酸，该片段在限制酶 Alf III 和 Nde I 作用下具有种特异性的酶切模式。所有 4 种标记在所有测试的苎麻品系上都得到了一致的结果。

结论

本研究证明了 RAPD、SCAR、SNP 和 CAPS 标记在快速鉴定两种密切相关的苎麻种方面的应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d465/3004904/b63343b28eef/1749-8546-5-40-6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d465/3004904/63f0c1766831/1749-8546-5-40-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d465/3004904/78ecac72cd03/1749-8546-5-40-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d465/3004904/a87bcf84246c/1749-8546-5-40-3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d465/3004904/82ee652e3839/1749-8546-5-40-4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d465/3004904/e792b725964f/1749-8546-5-40-5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d465/3004904/b63343b28eef/1749-8546-5-40-6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d465/3004904/63f0c1766831/1749-8546-5-40-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d465/3004904/78ecac72cd03/1749-8546-5-40-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d465/3004904/a87bcf84246c/1749-8546-5-40-3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d465/3004904/82ee652e3839/1749-8546-5-40-4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d465/3004904/e792b725964f/1749-8546-5-40-5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d465/3004904/b63343b28eef/1749-8546-5-40-6.jpg

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