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Volume 38 Issue 1
Mar.  2020
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Article Navigation >  Journal of Pharmaceutical Practice and Service  > 2020  >  38(1): 42-46,62

JIANG Shenggui, DONG Zhiying, LI Qing, ZHAO Yingkui, HUANG Doudou, SUN Lianna. Establish and optimization of inter-simple sequence repeat PCR reaction system of Gnaphalium affine[J]. Journal of Pharmaceutical Practice and Service, 2020, 38(1): 42-46,62. doi: 10.3969/j.issn.1006-0111.201909048
Citation: JIANG Shenggui, DONG Zhiying, LI Qing, ZHAO Yingkui, HUANG Doudou, SUN Lianna. Establish and optimization of inter-simple sequence repeat PCR reaction system of Gnaphalium affine[J]. Journal of Pharmaceutical Practice and Service, 2020, 38(1): 42-46,62. doi: 10.3969/j.issn.1006-0111.201909048
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Establish and optimization of inter-simple sequence repeat PCR reaction system of Gnaphalium affine

doi: 10.3969/j.issn.1006-0111.201909048
  • 1.

    School of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China

  • 2.

    School of Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China

  • 3.

    Department of Pharmacy, Changzheng Hospital, Navy Medical University, Shanghai 200003, China

  • 4.

    School of Pharmacy, Navy Medical University, Shanghai 200433, China

  • 5.

    School of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China;School of Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China

  • Received Date: 2019-09-12
  • Rev Recd Date: 2019-10-21
  • Objective To provide the experimental basis for the subsequent genetic diversity research through establishing and optimizing the inter-simple sequence repeat PCR (ISSR-PCR) reaction system of Gnaphalium affine. Methods The single-factor experimental method and full experimental method were used to optimize the ISSR-PCR reaction system of Gnaphalium affine. Under the optimal system, after screening primers and corresponding annealing temperatures, the systematic feasibility was verified. Results The optimal ISSR-PCR reaction system was consisted of 10 μl Premix Taq DNA polymerase, 0.3 μmol/L primer, 10 ng DNA template, and sterilized water added to 20 μl. Finally, 10 primers were screened from 100 universal primers, and verification results indicated the system had high stability, good reproducibility, and the selected primers had good polymorphism. Conclusion The ISSR-PCR amplification system of Gnaphalium affine was established for the first time and the primers with appropriate annealing temperatures were filtered out, which provided a reference for the subsequent genetic diversity research of Gnaphalium affine.
  • [1] LI J L, HUANG D D, CHEN W S, et al. Two new phenolic glycosides from Gnaphalium affine D. Don and their anti-complementary activity[J]. Molecules,2013,18(7):7751-7760
    [2] ZHANG H J, LI L N, ZHOU J, et al. Effects of Gnaphalium affine D. Don on hyperuricemia and acute gouty arthritis[J]. J Ethnopharmacol,2017,203:304-311
    [3] LIN Y, LIU P G, LIANG W Q, et al. Luteolin-4'-O-glucoside and its aglycone, two major flavones of Gnaphalium affine D. Don, resist hyperuricemia and acute gouty arthritis activity in animal models[J]. Phytomedicine,2018,41:54-61
    [4] SEONG Y A, HWANG D, KIM G D. The anti-inflammatory effect of Gnaphalium affine through inhibition of NF-κB and MAPK in lipopolysaccharide-stimulated RAW264.7 cells and analysis of its phytochemical components[J]. Cell Biochem Biophys,2016,74(3):407-417
    [5] LIN W Q, XIE J X, WU X M, et al. Inhibition of xanthine oxidase activity by Gnaphalium affine extract[J]. Chin Med Sci J,2014,29(4):225-230
    [6] ZHANG W, WU C Z, FAN S Y. Chemical constituents from Gnaphalium affine and their xanthine oxidase inhibitory activity[J]. Chin J Nat Med,2018,16(5):347-353
    [7] 高俊斌, 王璇, 陈燕红, 等. HPLC法同时测定鼠曲草中7种成分[J]. 中成药, 2018, 40(5):1116-1119
    [8] GOULÃO L, OLIVEIRA C M. Molecular characterisation of cultivars of apple (Malus×domestica Borkh.) using microsatellite (SSR and ISSR) markers[J]. Euphytica,2001,122(1):81-89
    [9] ZIETKIEWICZ E, RAFALSKI A, LABUDA D. Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification[J]. Genomics,1994,20(2):176-183
    [10] 任梦云, 陈彦君, 张盾, 等. 山莨菪ISSR-PCR反应体系的建立与优化[J]. 分子植物育种, 2017, 15(12):5006-5014
    [11] 任风鸣, 胡开治, 刘燕琴, 等. 传统中药金钱草ISSR-PCR反应体系的正交优化研究[J]. 中国中药杂志, 2014, 39(12):2233-2238
    [12] 唐辉, 陈宗游, 史艳财, 等. 正交设计优化地枫皮ISSR-PCR反应体系[J]. 中草药, 2013, 44(5):610-615
    [13] 张福生, 郭顺星. 金线莲ISSR反应体系的建立与优化[J]. 中草药, 2011, 42(1):137-142
    [14] 穆立蔷, 刘赢男, 冯富娟, 等. 紫椴ISSR-PCR反应体系的建立与优化[J]. 林业科学, 2006, 42(6):26-31
    [15] CUI C J, LI Y, LIU Y L, et al. Determination of genetic diversity among Saccharina germplasm using ISSR and RAPD markers[J]. C R Biol,2017,340(2):76-86
    [16] WU W F, CHEN F X, YEH K, et al. ISSR analysis of genetic diversity and structure of plum varieties cultivated in southern China[J]. Biology (Basel),2018,8(1):E2
    [17] 刘蕤, 杨际双. 菊属11个野生种和12个栽培品种遗传关系的ISSR分析[J]. 基因组学与应用生物学, 2009, 28(5):874-882
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Establish and optimization of inter-simple sequence repeat PCR reaction system of Gnaphalium affine

doi: 10.3969/j.issn.1006-0111.201909048
  • 1. School of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China
  • 2. School of Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
  • 3. Department of Pharmacy, Changzheng Hospital, Navy Medical University, Shanghai 200003, China
  • 4. School of Pharmacy, Navy Medical University, Shanghai 200433, China
  • 5. School of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China;School of Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
  • Received Date: 2019-09-12
  • Rev Recd Date: 2019-10-21

Abstract: Objective To provide the experimental basis for the subsequent genetic diversity research through establishing and optimizing the inter-simple sequence repeat PCR (ISSR-PCR) reaction system of Gnaphalium affine. Methods The single-factor experimental method and full experimental method were used to optimize the ISSR-PCR reaction system of Gnaphalium affine. Under the optimal system, after screening primers and corresponding annealing temperatures, the systematic feasibility was verified. Results The optimal ISSR-PCR reaction system was consisted of 10 μl Premix Taq DNA polymerase, 0.3 μmol/L primer, 10 ng DNA template, and sterilized water added to 20 μl. Finally, 10 primers were screened from 100 universal primers, and verification results indicated the system had high stability, good reproducibility, and the selected primers had good polymorphism. Conclusion The ISSR-PCR amplification system of Gnaphalium affine was established for the first time and the primers with appropriate annealing temperatures were filtered out, which provided a reference for the subsequent genetic diversity research of Gnaphalium affine.

JIANG Shenggui, DONG Zhiying, LI Qing, ZHAO Yingkui, HUANG Doudou, SUN Lianna. Establish and optimization of inter-simple sequence repeat PCR reaction system of Gnaphalium affine[J]. Journal of Pharmaceutical Practice and Service, 2020, 38(1): 42-46,62. doi: 10.3969/j.issn.1006-0111.201909048
Citation: JIANG Shenggui, DONG Zhiying, LI Qing, ZHAO Yingkui, HUANG Doudou, SUN Lianna. Establish and optimization of inter-simple sequence repeat PCR reaction system of Gnaphalium affine[J]. Journal of Pharmaceutical Practice and Service, 2020, 38(1): 42-46,62. doi: 10.3969/j.issn.1006-0111.201909048
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