Full Text:   <1981>

Summary:  <1470>

Suppl. Mater.: 

CLC number: 

On-line Access: 2021-02-07

Received: 2020-08-10

Revision Accepted: 2020-10-08

Crosschecked: 2021-01-06

Cited: 0

Clicked: 3190

Citations:  Bibtex RefMan EndNote GB/T7714


Huixia SHOU


-   Go to

Article info.
Open peer comments

Journal of Zhejiang University SCIENCE B 2021 Vol.22 No.2 P.99-111


Evaluation and application of an efficient plant DNA extraction protocol for laboratory and field testing

Author(s):  Qi WANG, Xiaoxia SHEN, Tian QIU, Wei WU, Lin LI, Zhi'an WANG, Huixia SHOU

Affiliation(s):  State Key Laboratory of Plant Physiology and Biochemistry, College of Life Sciences, Zhejiang University, Hangzhou 310058, China; more

Corresponding email(s):   huixia@zju.edu.cn

Key Words:  DNA extraction, Cellulose filter paper, Field test, Genetically modified (GM) crops, Chinese medicine ultra-fine powder, Rapid molecular identification

Qi WANG, Xiaoxia SHEN, Tian QIU, Wei WU, Lin LI, Zhi'an WANG, Huixia SHOU. Evaluation and application of an efficient plant DNA extraction protocol for laboratory and field testing[J]. Journal of Zhejiang University Science B, 2021, 22(2): 99-111.

@article{title="Evaluation and application of an efficient plant DNA extraction protocol for laboratory and field testing",
author="Qi WANG, Xiaoxia SHEN, Tian QIU, Wei WU, Lin LI, Zhi'an WANG, Huixia SHOU",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Evaluation and application of an efficient plant DNA extraction protocol for laboratory and field testing
%A Xiaoxia SHEN
%A Tian QIU
%A Wei WU
%A Lin LI
%A Zhi'an WANG
%A Huixia SHOU
%J Journal of Zhejiang University SCIENCE B
%V 22
%N 2
%P 99-111
%@ 1673-1581
%D 2021
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B2000465

T1 - Evaluation and application of an efficient plant DNA extraction protocol for laboratory and field testing
A1 - Qi WANG
A1 - Xiaoxia SHEN
A1 - Tian QIU
A1 - Wei WU
A1 - Lin LI
A1 - Zhi'an WANG
A1 - Huixia SHOU
J0 - Journal of Zhejiang University Science B
VL - 22
IS - 2
SP - 99
EP - 111
%@ 1673-1581
Y1 - 2021
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B2000465

Nucleic acids in plant tissue lysates can be captured quickly by a cellulose filter paper and prepared for amplification after a quick purification. In this study, a published filter paper strip method was modified by sticking the filter paper on a polyvinyl chloride resin (PVC) sheet. This modified method is named EZ-D, for EASY DNA extraction. Compared with the original cetyl trimethylammonium bromide (CTAB) method, DNA extracted by EZ-D is more efficient in polymerase chain reaction (PCR) amplification due to the more stable performance of the EZ-D stick. The EZ-D method is also faster, easier, and cheaper. PCR analyses showed that DNA extracted from several types of plant tissues by EZ-D was appropriate for specific identification of biological samples. A regular PCR reaction can detect the EZ-D-extracted DNA template at concentration as low as 0.1 ng/μL. Evaluation of the EZ-D showed that DNA extracts could be successfully amplified by PCR reaction for DNA fragments up to 3000 bp in length and up to 80% in GC content. EZ-D was successfully used for DNA extraction from a variety of plant species and plant tissues. Moreover, when EZ-D was combined with the loop-mediated isothermal amplification (LAMP) method, DNA identification of biological samples could be achieved without the need for specialized equipment. As an optimized DNA purification method, EZ-D shows great advantages in application and can be used widely in laboratories where equipment is limited and rapid results are required.


目的:本研究旨在开发一种优化的滤纸法快速纯化DNA技术--EZ-D(EASY DNA),并将EZ-D与聚合酶链式反应(PCR)/环介导等温扩增(LAMP)技术相结合应用于实验室或室外不同类型植物材料的目的基因的快速鉴定,同时对EZ-D方法的DNA提取效率和所提取DNA的完整性和可扩增能力进行评价。


Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1]Chen SL, Pang XH, Song JY, et al., 2014. A renaissance in herbal medicine identification: from morphology to DNA. Biotechnol Adv, 32(7):1237-1244.

[2]Cheng W, Xia ZJ, Feng XZ, et al., 2016. A rapid and nondestructive method for soybean DNA extraction and its application. Chin Bull Bot, 51(1):68-73 (in Chinese).

[3]Fan YH, Wang PL, Fan XS, et al., 2016. Research on application status of Chinese herbal decoction pieces based on clinical survey. China J Chin Mater Med, 41(15):2927-2931 (in Chinese).

[4]Goto M, Honda E, Ogura A, et al., 2009. Colorimetric detection of loop-mediated isothermal amplification reaction by using hydroxy naphthol blue. Short Tech Rep, 46(3):167-172.

[5]Hou SM, Duan JQ, Liang XN, et al., 2005. Optimized CTAB protocol for extracting the total DNA of ramie. Acta Bot Boreali-Occid Sin, 25(11):2193-2197 (in Chinese).

[6]Jiang C, Huang LQ, Yuan Y, et al., 2013. Rapid extraction of DNA from Chinese medicinal materials by alkaline lysis. Chin J Pharm Anal, 33(7):1081-1090 (in Chinese).

[7]Kiddle G, Hardinge P, Buttigieg N, et al., 2012. GMO detection using a bioluminescent real time reporter (BART) of loop mediated isothermal amplification (LAMP) suitable for field use. BMC Biotechnol, 12:15.

[8]Lan QK, Zhao X, Chen R, et al., 2019. Authentication of Fritillaria cirrhosa based on loop-mediate isothermal amplification. Chin J Pharm Anal, 39(3):551-556 (in Chinese).

[9]Li HJ, Zhao YC, Li LH, 2017. The research progress of Chinese medicine superfine powder quality standard. Mod Anim Husbandry, 1(2):30-32 (in Chinese).

[10]Li JJ, Xiong C, Liu Y, et al., 2016. Loop-mediated isothermal amplification (LAMP): emergence as an alternative technology for herbal medicine identification. Front Plant Sci, 7:1956.

[11]Li M, Zhao X, 2012. RAPD analysis of three species of Fritillaria in southern China. J Zhejiang Univ Technol, 40(6):634-638 (in Chinese).

[12]Li M, Huang LM, Zhao X, et al., 2014. Specific PCR identification of Fritillaria thunbergii . Chin Tradit Herb Drugs, 45(12):1754-1757 (in Chinese).

[13]Liesenfeld O, Lehman L, Hunfeld KP, et al., 2014. Molecular diagnosis of sepsis: new aspects and recent developments. Eur J Microbiol Immunol, 4(1):1-25.

[14]Liu H, Wang JB, Li P, et al., 2020. Rapid detection of P‒35S and T-nos in genetically modified organisms by recombinase polymerase amplification combined with a lateral flow strip. Food Control, 107:106775.

[15]Liu XX, Li J, Zhang YB, et al., 2019. Rapid identification of Fritillariae Cirrhosae Bulbus by polymerase chain reaction (PCR) method. Chin J Pharm Anal, 39(10):1844-1851 (in Chinese).

[16]Lu LH, Han Q, Li L, et al., 2014. Establishment of an efficient transformation protocol for soybean using glyphosate as selective agent and the development of glyphosate-tolerant transgenic soybean lines. Sci Sin Vitae, 44(4):406-415.

[17]Moeller JR, Moehn NR, Waller DM, et al., 2014. Paramagnetic cellulose DNA isolation improves DNA yield and quality among diverse plant taxa. Appl Plant Sci, 2(10):1400048.

[18]Murray MG, Thompson WF, 1980. Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res, 8(19):4321-4326.

[19]Notomi T, Okayama H, Masubuchi H, et al., 2000. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res, 28(12):e63.

[20]Paul R, Saville AC, Hansel JC, et al., 2019. Extraction of plant DNA by microneedle patch for rapid detection of plant diseases. ACS Nano, 13(6):6540-6549.

[21]Peterson DG, Boehm KS, Stack SM, 1997. Isolation of milligram quantities of nuclear DNA from tomato (Lycopersicon esculentum), a plant containing high levels of polyphenolic compounds. Plant Mol Biol Rep, 15(2):148-153.

[22]Ren LZ, Zhang CB, Zhao HK, et al., 2012. An improved method to rapid and high-quality of genomic DNA extraction from soybean. Chin Agric Sci Bull, 28(9):38-41 (in Chinese).

[23]Tomita N, Mori Y, Kanda H, et al., 2008. Loop-mediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products. Nat Protoc, 3(5):877-882.

[24]Tu JM, Zhang GA, Datta K, et al., 2000. Field performance of transgenic elite commercial hybrid rice expressing Bacillus thuringiensis δ-endotoxin. Nat Biotechnol, 18(10):1101-1104.

[25]Varma A, Padh H, Shrivastava N, 2007. Plant genomic DNA isolation: an art or a science. Biotechnol J, 2(3):386-392.

[26]Wang HM, Li YX, Weng HL, et al., 2012. Extracted Larix gmelinii DNA with improved DNA extraction kits. J Anhui Agric Sci, 40(35):17029-17030, 17041 (in Chinese).

[27]Wang SD, Yokosho Y, Guo RZ, et al., 2019. The soybean sugar transporter GmSWEET15 mediates sucrose export from endosperm to early embryo. Plant Physiol, 180(4):2133-2141.

[28]Wang SD, Li L, Ying YH, et al., 2020. A transcription factor OsbHLH156 regulates Strategy II iron acquisition through localising IRO2 to the nucleus in rice. New Phytol, 225(3):1247-1260.

[29]Wang SS, Wang F, Zhang H, et al., 2020. Rapid extraction of DNA from Chinese herbal medicines by filter paper method. Mod Chin Med, 22(2):213-218 (in Chinese).

[30]Wu YY, Dai DY, Cai CM, 2015. A modified UREA protocol for soybean DNA extraction. Soybean Sci, 34(1):112-115 (in Chinese).

[31]Yang L, Wu WR, Fu F, et al., 2019. Exploration and application of a new method for rapid extraction of DNA from Chinese medicinal materials. Chin Tradit Herbal Drugs, 50(2):502-509 (in Chinese).

[32]Yu ZX, Wei CJ, Li YM, et al., 2016. Study on DNA extraction method of genetically modified soybean. Guangdong Chem Ind, 43(18):23-24 (in Chinese).

[33]Zhang JJ, Xu H, Zhao SJ, 2017. Rapid identification of medicinal herbs through plant Direct-PCR. Acta Pharm Sin, 52(11):1763-1769 (in Chinese).

[34]Zhang LF, Li SG, Han SY, et al., 2013. Establishment of larch transgenic breeding system and its industrial application. Biotechnol Bus, (3):7-11 (in Chinese).

[35]Zhang XZ, Lowe SB, Gooding JJ, 2014. Brief review of monitoring methods for loop-mediated isothermal amplification (LAMP). Biosens Bioelectron, 61:491-499.

[36]Zhao ZP, Yan Y, Zheng MM, et al., 2019. Genetic evaluation of Gentiana rigescens based on the chloroplast psbA-trnH, trnL-trnF sequence polymorphism. Lishizhen Med Mater Med Res, 30(5):1203-1206 (in Chinese).

[37]Zou YP, Mason MG, Wang YL, et al., 2017. Nucleic acid purification from plants, animals and microbes in under 30 seconds. PLoS Biol, 15(11):e2003916.

Open peer comments: Debate/Discuss/Question/Opinion


Please provide your name, email address and a comment

Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - 2024 Journal of Zhejiang University-SCIENCE