Rapid Prediction of Contents of Sibiricose A5 and Sibiricose A6 in Polygalae Radix with ISSR Markers

WANG Hai-hua; WANG Dan-dan; CHEN Tong-yao; ZHANG Fu-sheng; QIN Xue-mei; MA Cun-gen

doi:10.13422/j.cnki.syfjx.20182106

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Rapid Prediction of Contents of Sibiricose A5 and Sibiricose A6 in Polygalae Radix with ISSR Markers

更新时间：2024-09-23

- Rapid Prediction of Contents of Sibiricose A5 and Sibiricose A6 in Polygalae Radix with ISSR Markers
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 24, Pages: 38-44(2018)
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- DOI：10.13422/j.cnki.syfjx.20182106
  CLC： R284.1
- Published：2018
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WANG Hai-hua, WANG Dan-dan, CHEN Tong-yao, et al. Rapid Prediction of Contents of Sibiricose A5 and Sibiricose A6 in Polygalae Radix with ISSR Markers[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(24): 38-44.
DOI：

WANG Hai-hua, WANG Dan-dan, CHEN Tong-yao, et al. Rapid Prediction of Contents of Sibiricose A5 and Sibiricose A6 in Polygalae Radix with ISSR Markers[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(24): 38-44. DOI： 10.13422/j.cnki.syfjx.20182106.

摘要

目的：优化远志简单重复序列区间（ISSR）扩增多态性反应体系，设计一对聚合酶链式反应（PCR）引物用于快速预测远志药材中sibiricose A5和sibiricoseA6的含量高低。方法：采用UPLC测定远志药材中sibiricose A5与sibiricose A6的含量，通过单因素试验优化远志ISSR反应体系，利用ISSR分子标记技术筛选出与sibiricose A5和sibiricose A6含量相关的特异性DNA片段，针对此片段设计特异性PCR引物。结果： 7批远志药材中sibiricose A5和sibiricose A6的含量差异较大，变化范围分别为0.895 4~3.227 6，0.644 2~2.057 8 mg·g-1。对筛选出的与sibiricose A5和sibiricose A6含量相关片段（890-8）进行克隆测序后，设计出了1对特异性引物（YZ-SA），可用于快速预测远志药材中sibiricose A5和sibiricose A6的含量高低。结论：优化后的远志ISSR反应体系具有标记位点清晰、检测多态性能力强等特点，设计出的特异性引物可用于快速预测远志药材中sibiricose A5和sibiricose A6的含量高低。

Abstract

Objective: The inter-simple sequence repeat(ISSR) reaction system of Polygalae Radix was optimized and a pair of polymerase chain reaction(PCR) primers was designed to rapidly predict the contents of sibiricose A5 and sibiricose A6 in Polygalae Radix. Method: The contents of sibiricose A5 and sibiricose A6 were determined by UPLC

and the ISSR reaction system of Polygalae Radix was optimized by single factor tests.The specific DNA fragments related to sibiricose A5 and sibiricose A6 were screened by ISSR

and a pair of specific PCR primers was designed for the fragments. Result: The contents of sibiricose A5 and sibiricose A6 were significantly different in 7 batches of Polygalae Radix

and the variation range of these two substances was 0.895 4-3.227 6

0.644 2-2.057 8 mg·g-1

respectively.After cloning and sequencing of 890-8 related to the contents of sibiricose A5 and sibiricose A6

a pair of specific primers (YZ-SA) was designed for rapidly predicting the contents of sibiricose A5 and sibiricose A6 in Polygalae Radix. Conclusion: The ISSR-PCR system established for Polygalae Radix shows the characters of clear marker site

reliable abundant polymorphisms and other characteristics.The specific PCR primers can be used to rapidly predict the contents of sibiricose A5 and sibiricose A6 in Polygalae Radix.

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