Application of Digital PCR in Plant Research

doi:10.16420/j.issn.0513-353x.2023-0971

Acta Horticulturae Sinica ›› 2024, Vol. 51 ›› Issue (12): 2977-2988.doi: 10.16420/j.issn.0513-353x.2023-0971

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Application of Digital PCR in Plant Research

GAO Yuan(), MA Yuehua, ZHANG Panpan, SHU Sheng, CHEN Sumei

College of Horticulture，Nanjing Agricultural University，Nanjing 210095，China

Received:2024-03-10 Revised:2024-09-03 Online:2024-12-25 Published:2024-12-13
Contact: GAO Yuan

Abstract

Abstract:

Digital PCR（dPCR）is a newly emerged technology to quantify the absolute nucleic acid content with high-sensitivity in recent years. It can achieve absolute quantification of the initial concentration of the sample without the need to rely on reference or standard curve，with characteristics of high sensitivity，specificity，and accuracy. This article discusses the basic principle of dPCR，and reviews its applications in the detection of genetically modified plants，plant pathogen diagnosis，copy number variation，gene expression research，and further perspectives for the application of dPCR in plants.

Key words: digital PCR, plant research, genetically modified plants, plant pathogen diagnosis, absolute quantification

GAO Yuan, MA Yuehua, ZHANG Panpan, SHU Sheng, CHEN Sumei. Application of Digital PCR in Plant Research[J]. Acta Horticulturae Sinica, 2024, 51(12): 2977-2988.

Figures/Tables 2

References 79

[1]	Bahder B W, Helmick E E, Mou D F, Harrison N A, Davis R. 2018. Digital PCR technology for detection of palm-infecting phytoplasmas belonging to group 16SrIV that occur in Florida. Plant Dis, 102 (5):1008-1014. doi: 10.1094/PDIS-06-17-0787-RE pmid: 30673395
[2]	Barone V, Bertoldo G, Magro F, Broccanello C, Puglisi I, Baglieri A, Cagnin M, Concheri G, Squartini A, Pizzeghello D. 2019. Molecular and morphological changes induced by leonardite-based biostimulant in Beta vulgaris L. Plants, 8 (6):181.
[3]	Barrett-Manako K, Andersen M, Martínez-Sánchez M, Jenkins H, Hunter S, Reese-George J, Montefiori M, Wohlers M, Rikkerink E, Templeton M, Nardozza S. 2021. Real-time PCR and droplet digital PCR are accurate and reliable methods to quantify Pseudomonas syringae pv. actinidiae Biovar 3 in kiwifruit infected plantlets. Plant Dis, 105 (6):1748-1757.
[4]	Becker-André M, Hahlbrock K. 1989. Absolute mRNA quantification using the polymerase chain reaction(PCR). A novel approach by a PCR aided transcript titration assay(PATTY). Nucleic Acids Res, 17 (22):9437-9446. pmid: 2479917
[5]	Belide S, Vanhercke T, Petrie J R, Singh S P. 2017. Robust genetic transformation of sorghum(Sorghum bicolor L.)using differentiating embryogenic callus induced from immature embryos. Plant Methods,13:109.
[6]	Boubegtitene A, Merret R. 2022. Monitoring mRNA half-life in Arabidopsis using droplet digital PCR. Plants(Basel), 11 (19):2616.
[7]	Broothaerts W, Beaz Hidalgo R, Buttinger G, Seghers J, Maretti M, Robouch P, Corbisier P. 2023. Proficiency of European GMO control laboratories to quantify MON89788 soybean in a meat pâté matrix. Food Control,145:109454.
[8]	Carrara I, Terzi V, Ghizzoni R, Delbono S, Tumino G, Crespan M, Gardiman M, Francia E, Morcia C. 2023. A molecular toolbox to identify and quantify grape varieties:on the trace of“Glera”. Foods, 12 (16):3091.
[9]	Chiodi C, Concheri G, Squartini A, Ravi S, Broccanello C, Moro M, Stevanato P. 2021. Quantification of rhizomania virus by automated RNA isolation and PCR based methods in sugar beet. Virus Disease, 32 (1):161-166.
[10]	Collier R, Dasgupta K, Xing Y P, Hernandez B T, Shao M, Rohozinski D, Kovak E, Lin J, de Oliveira M L P, Stover E, McCue K F, Harmon F G, Blechl A, Thomson J G, Thilmony R. 2017. Accurate measurement of transgene copy number in crop plants using droplet digital PCR. Plant J, 90 (5):1014-1025.
[11]	Corbisier P, Bhat S, Partis L, Xie V R, Emslie K R. 2010. Absolute quantification of genetically modified MON810 maize(Zea mays L.) by digital polymerase chain reaction. Anal Bioanal Chem, 396 (6):2143-2150. doi: 10.1007/s00216-009-3200-3 pmid: 19816678
[12]	Cottenet G, Blancpain C, Chuah P F. 2019. Performance assessment of digital PCR for the quantification of GM-maize and GM-soya events. Anal Bioanal Chem, 411 (11):2461-2469. doi: 10.1007/s00216-019-01692-7 pmid: 30810790
[13]	Cristiano D, Peruzy M F, Aponte M, Mancusi A, Proroga Y T R, Capuano F, Murru N. 2021. Comparison of droplet digital PCR vs real-time PCR for Yersinia enterocolitica detection in vegetables. Int J Food Microbiol,354:109321.
[14]	Darvishi N, Alavi S M, Hijazi M. 2021. CRISPR-mediated mutation in cinnamoyl-CoA reductase 4 in allohexaploid oilseed crop Camelina sativa,revealed its pivotal role in resistance against Sclerotinia sclerotiorum. Iran J Biotechnol, 19 (4):e2768. doi: 10.30498/ijb.2021.230722.2768 pmid: 35350644
[15]	Demeke T, Gräfenhan T, Holigroski M, Fernando U, Bamforth J, Lee S J. 2014. Assessment of droplet digital PCR for absolute quantification of genetically engineered OXY 235 canola and DP305423 soybean samples. Food Control,46:470-474.
[16]	Demeke T, Lee S J, Eng M. 2022. Increasing the efficiency of canola and soybean GMO detection and quantification using multiplex droplet digital PCR. Biology(Basel), 11 (2):201.
[17]	Dobnik D, Spilsberg B, Bogožalec Košir A, Holst-Jensen A, Žel J. 2015. Multiplex quantification of 12 European Union authorized genetically modified maize lines with droplet digital polymerase chain reaction. Anal Chem, 87 (16):8218-8226. doi: 10.1021/acs.analchem.5b01208 pmid: 26169291
[18]	Dong X, Gao D, Dong J, Chen W, Li Z, Wang J, Liu J. 2020. Mass ratio quantitative detection for kidney bean in lotus seed paste using duplex droplet digital PCR and chip digital PCR. Analytical and Bioanalytical Chemistry, 412 (7):1701-1707. doi: 10.1007/s00216-020-02410-4 pmid: 31965247
[19]	Dreo T, Pirc M, Ramšak Ž, Pavšič J, Milavec M, Žel J, Gruden K. 2014. Optimising droplet digital PCR analysis approaches for detection and quantification of bacteria:a case study of fire blight and potato brown rot. Analytical and Bioanalytical Chemistry, 406 (26):6513-6528.
[20]	Dressman D, Yan H, Traverso G, Kinzler K W, Vogelstein B. 2003. Transforming single DNA molecules into fluorescent magnetic particles for detection and enumeration of genetic variations. Proceedings of the National Academy of Sciences of the United States of America, 100 (15):8817-8822.
[21]	Du Y, Wang M, Zou L, Long M, Yang Y, Zhang Y, Liang X. 2021. Quantitative detection and monitoring of Colletotrichum siamense in rubber trees using real-time PCR. Plant Dis, 105 (10):2861-2866.
[22]	Duan K, Zhao Y, Li Z, Zou X, Yang J, Guo C, Chen S, Yang X, Gao Q. 2021. A strategy for the production and molecular validation of agrobacterium-mediated intragenic octoploid strawberry. Plants(Basel), 10 (11):2229.
[23]	Gao D, Abdullah S, Baldwin T, Caspersen A, Williams E, Carlson A, Petersen M, Hu G, Klos K E, Bregitzer P. 2024. Agrobacterium-mediated transfer of the Fusarium graminearum Tri 6 gene into barley using mature seed-derived shoot tips as explants. Plant Cell Rep, 43 (2):40.
[24]	Gao R, Feyissa B A, Croft M, Hannoufa A. 2018. Gene editing by CRISPR/Cas 9 in the obligatory outcrossing Medicago sativa. Planta, 247 (4):1043-1050.
[25]	Głowacka K, Kromdijk J, Leonelli L, Niyogi K K, Clemente T E, Long S P. 2016. An evaluation of new and established methods to determine T-DNA copy number and homozygosity in transgenic plants. Plant Cell Environ, 39 (4):908-917.
[26]	Hansen S J Z, Morovic W, DeMeules M, Stahl B, Sindelar C W. 2018. Absolute enumeration of probiotic strains Lactobacillus acidophilus NCFM^®and Bifidobacterium animalis subsp. lactis Bl-04^® via chip-based digital PCR. Front Microbiol,9:704.
[27]	Hansen S J Z, Tang P, Kiefer A, Galles K, Wong C, Morovic W. 2019. Droplet digital PCR is an improved alternative method for high-quality enumeration of viable probiotic strains. Front Microbiol,10:3025.
[28]	Higuchi R, Fockler C, Dollinger G, Watson R. 1993. Kinetic PCR analysis:real-time monitoring of DNA amplification reactions. Biotechnology (N Y), 11 (9):1026-1030.
[29]	Hrycan J, Theilmann J, Mahovlic A, Boulé J, Úrbez-Torres J R. 2023. Health status of ready-to-plant grapevine nursery material in Canada regarding young vine decline fungi. Plant Dis, 107 (12):3708-3717. doi: 10.1094/PDIS-05-23-0900-SR pmid: 37436216
[30]	Hua S S T, Palumbo J D, Parfitt D E, Sarreal S B L, O’Keeffe T L. 2018. Development of a droplet digital PCR assay for population analysis of aflatoxigenic and atoxigenic Aspergillus flavus mixtures in soil. Mycotoxin Research, 34 (3):187-194.
[31]	Huggett J F. 2020. The Digital MIQE guidelines update:minimum information for publication of quantitative digital PCR experiments for 2020. Clin Chem, 66 (8):1012-1029. doi: 10.1093/clinchem/hvaa125 pmid: 32746458
[32]	Jia R, Zhao H, Huang J, Kong H, Zhang Y, Guo J, Huang Q, Guo Y, Wei Q, Zuo J, Zhu Y J, Peng M, Guo A. 2017. Use of RNAi technology to develop a PRSV-resistant transgenic papaya. Sci Rep, 7 (1):12636. doi: 10.1038/s41598-017-13049-0 pmid: 28974762
[33]	Karppinen K, Avetisyan A, Hykkerud A L, Jaakola L. 2022. A dPCR Method for quantitative authentication of wild lingonberry(Vaccinium vitis-idaea)versus cultivated American cranberry(V. macrocarpon). Foods, 11 (10):1476.
[34]	Keremane M L, McCollum T G, Roose M L, Lee R F, Ramadugu C. 2021. An improved reference gene for detection of“Candidatus Liberibacter asiaticus”associated with citrus Huanglongbing by qPCR and digital droplet PCR assays. Plants (Basel), 10 (10):2111.
[35]	Knight N L, Moslemi A, Begum F, Dodhia K N, Covarelli L, Hills A L, Lopez-Ruiz F J. 2022. Detection of Ramularia collo-cygni from barley in Australia using triplex quantitative and droplet digital PCR. Pest Manag Sci, 78 (4):1367-1376.
[36]	Lancíková V, Hricová A. 2020. Digital absolute gene expression analysis of essential starch-related genes in a radiation developed Amaranthus cruentus L. variety in comparison with real-time PCR. Plants, 9 (8):966.
[37]	Lee H, Cho I, Ju H, Jeong R. 2021. Development of a reverse transcription droplet digital PCR assay for sensitive detection of peach latent mosaic viroid. Mol Cell Probes,58:101746.
[38]	Leichtfried T, Reisenzein H, Steinkellner S, Gottsberger R A. 2020. Transmission studies of the newly described apple chlorotic fruit spot viroid using a combined RT-qPCR and droplet digital PCR approach. Arch Virol, 165 (11):2665-2671.
[39]	Li H, Wei C, Meng Y, Fan R, Zhao W, Wang X, Yu X, Laroche A, Kang Z, Liu D. 2020. Identification and expression analysis of some wheat F-box subfamilies during plant development and infection by Puccinia triticina. Plant Physiol Biochem,155:535-548.
[40]	Li J, Zhai S, Gao H, Xiao F, Li Y, Wu G, Wu Y. 2021. Development and assessment of a duplex droplet digital PCR method for quantification of GM rice Kemingdao. Anal Bioanal Chem, 413 (16):4341-4351. doi: 10.1007/s00216-021-03390-9 pmid: 34023912
[41]	Low H, Chan S, Soo G, Ling B, Tan E. 2017. Clarity™ digital PCR system:a novel platform for absolute quantification of nucleic acids. Anal Bioanal Chem, 409 (7):1869-1875.
[42]	Lu Y, Zhang H, Zhao Z, Wen C, Wu P, Song S, Yu S, Luo L, Xu X. 2020. Application of droplet digital PCR in detection of seed-transmitted pathogen Acidovorax citrulli. Journal of Integrative Agriculture, 19 (2):561-569.
[43]	Luigi M, Manglli A, Corrado C L, Tiberini A, Costantini E, Ferretti L, Tomassoli L, Bertin S. 2023. Development,validation,and application of reverse transcription real-time and droplet digital PCR assays for the detection of the potyviruses watermelon mosaic virus and zucchini yellow mosaic virus in cucurbits. Plants(Basel), 12 (12):2364.
[44]	McCord Per H. 2016. Using droplet digital PCR(ddPCR)to detect copy number variation in sugarcane,a high-level polyploid. Euphytica, 209 (2):439-448.
[45]	McCue K F, Gardner E, Chan R, Thilmony R, Thomson J. 2019. Transgene stacking in potato using the GAANTRY system. BMC Res Notes, 12 (1):457. doi: 10.1186/s13104-019-4493-8 pmid: 31345264
[46]	Morcia C, Bergami R, Scaramagli S, Ghizzoni R, Carnevali P, Terzi V. 2020. A chip digital PCR assay for quantification of common wheat contamination in pasta production chain. Foods, 9 (7):911.
[47]	Morcia C, Tumino G, Raimondi S, Schneider A, Terzi V. 2021. Muscat flavor in grapevine:a digital PCR assay to track allelic variation in VvDXS Gene. Genes(Basel), 12 (5):747.
[48]	Mullis K B. 1990. The unusual origin of the polymerase chain reaction. Sci Am, 262 (4):56-61,64-55. pmid: 2315679
[49]	Murolo S, Moumni M, Mancini V, Allagui M B, Landi L, Romanazzi G. 2021. Detection and quantification of Stagonosporopsis cucurbitacearum in seeds of Cucurbita maxima using droplet digital polymerase chain reaction. Front Microbiol,12:764447.
[50]	Narancio R, John U, Mason J, Giraldo P, Spangenberg G. 2021. Digital PCR(dPCR)and qPCR mediated determination of transgene copy number in the forage legume white clover(Trifolium repens). Mol Biol Rep, 48 (4):3069-3077.
[51]	Notaguchi M, Higashiyama T, Suzuki T. 2015. Identification of mRNAs that move over long distances using an RNA-Seq analysis of Arabidopsis/Nicotiana benthamiana heterografts. Plant Cell Physiol, 56 (2):311-321.
[52]	Pandey B, Mallik I, Gudmestad N C. 2020. Development and application of a real-time reverse-transcription PCR and droplet digital PCR assays for the direct detection of potato mop top virus in soil. Phytopathology, 110 (1):58-67. doi: 10.1094/PHYTO-05-19-0185-FI pmid: 31448996
[53]	Pérez-López E, Rodríguez-Martínez D, Olivier C Y, Luna-Rodríguez M, Dumonceaux T J. 2017. Molecular diagnostic assays based on cpn 60 UT sequences reveal the geographic distribution of subgroup 16SrXIII-(A/I) I phytoplasma in Mexico. Sci Rep, 7 (1):950. doi: 10.1038/s41598-017-00895-1 pmid: 28424530
[54]	Philips A, Nowis K, Stelmaszczuk M, Jackowiak P, Podkowiński J, Handschuh L, Figlerowicz M. 2020. Expression landscape of circRNAs in Arabidopsis thaliana seedlings and adult tissues. Front Plant Sci,11:576581.
[55]	Quan P L, Sauzade M, Brouzes E. 2018. dPCR:a technology review. Sensors(Basel), 18 (4):1271.
[56]	Ren Z, Chen R, Muhae-Ud-Din G, Fang M, Li T, Yang Y, Chen W, Gao L. 2022. Development of real-time PCR and droplet digital PCR based marker for the detection of Tilletia caries inciting common bunt of wheat. Front Plant Sci,13:1031611.
[57]	Ruijter J M, Ramakers C, Hoogaars W M, Karlen Y, Bakker O, van den Hoff M J, Moorman A F. 2009. Amplification efficiency:linking baseline and bias in the analysis of quantitative PCR data. Nucleic Acids Res, 37 (6):e45.
[58]	Santander R D, Gašić K, Aćimović S G. 2022. Selective quantification of Erwinia amylovora live cells in pome fruit tree cankers by viability digital PCR. Methods Mol Biol,2536:231-249.
[59]	Santos C, Duarte S, Tedesco S, Fevereiro P, Costa R L. 2017. Expression profiling of Castanea genes during resistant and susceptible interactions with the oomycete pathogen Phytophthora cinnamomi reveal possible mechanisms of immunity. Frontiers in Plant Science,8:515.
[60]	Schmiderer C, Steinborn R, Novak J. 2023. Monoterpene synthases of three closely related sage species(Salvia officinalis,S. fruticosa and S. pomifera,Lamiaceae). Plant Physiology and Biochemistry,196:318-327.
[61]	Shen J, Shou W, Zhang Y, Yuan G, Zhao Y, Chen J, Havey M J. 2019a. Rare maternal and biparental transmission of the cucumber mitochondrial DNA reveals sorting of polymorphisms among progenies. Theor Appl Genet, 132 (4):1223-1233.
[62]	Shen J, Zhang Y, Havey M J, Shou W. 2019b. Copy numbers of mitochondrial genes change during melon leaf development and are lower than the numbers of mitochondria. Hortic Res,6:95.
[63]	Sykes P J, Neoh S H, Brisco M J, Hughes E, Condon J, Morley A A. 1992. Quantitation of targets for PCR by use of limiting dilution. Biotechniques, 13 (3):444-449. pmid: 1389177
[64]	Tan C, Fan D, Wang N, Wang F, Wang B, Zhu L, Guo Y. 2021. Applications of digital PCR in COVID-19 pandemic. View(Beijing), 2 (2):20200082.
[65]	Tellinghuisen J, Spiess A N. 2015. Bias and imprecision in analysis of real-time quantitative polymerase chain reaction data. Anal Chem, 87 (17):8925-8931. doi: 10.1021/acs.analchem.5b02057 pmid: 26235706
[66]	Tian Y, Fei J, Luo J, Chen L, Ye J, Du W, Yu C. 2023. Development of a reverse-transcription droplet digital PCR method for quantitative detection of Cucumber green mottle mosaic virus. Heliyon, 9 (2):e12643.
[67]	Vogelstein B, Kinzler K W. 1999. Digital PCR. Proceedings of the National Academy of Sciences of the United States of America, 96 (16):9236-9241.
[68]	Wang L, Tian Q, Zhou P, Zhao W, Sun X. 2022. Evaluation of droplet digital PCR for the detection of black canker disease in tomato. Plant Dis, 106 (2):395-405.
[69]	Wang S, Cui H, Chen M, Wu Y, Wang S. 2023. Quantitative PCR assays for the species-specific detection of Fusarium graminearum sensu stricto and Fusarium asiaticum in winter wheat growing regions in China. Int J Food Microbiol,387:110061.
[70]	Wu Y, Li J, Li X, Liang J, Li Y, Zeng X, Wu G. 2017. Copy number and zygosity determination of transgenic rapeseed by droplet digital PCR. Oil Crop Science, 1 (2):84-94.
[71]	Xie S, Yu H, Wang Q, Cheng Y, Ding T. 2020. Two rapid and sensitive methods based on TaqMan qPCR and droplet digital PCR assay for quantitative detection of Bacillus subtilis in rhizosphere. Journal of Applied Microbiology, 128 (2):518-527.
[72]	Xu X, Peng C, Wang X, Chen X, Wang Q, Xu J. 2016. Comparison of droplet digital PCR with quantitative real-time PCR for determination of zygosity in transgenic maize. Transgenic Res, 25 (6):855-864. pmid: 27632191
[73]	Yakimowski S B, Teitel Z, Caruso C M. 2021. Defence by duplication:The relation between phenotypic glyphosate resistance and EPSPS gene copy number variation in Amaranthus palmeri. Mol Ecol, 30 (21):5328-5342. doi: 10.1111/mec.16231 pmid: 34662479
[74]	Yang Shou, Jiang Feng, Liu Yan, Li Shiyao, Wang Mingqiu, Ma Yi, Lin Min, Zhang Li. 2017. Duplex digital droplet PCR for the determination of walnut-derived and soybean-derived ingredients in walnut protein drink. Food Science, 38 (16):280-286. (in Chinese)
	杨硕, 江丰, 刘艳, 李诗瑶, 王鸣秋, 马弋, 林敏, 张莉. 2017. 多重微滴式数字PCR定量检测市售核桃乳中核桃、大豆源性成分. 食品科学, 38 (16):280-286.
[75]	You Yinghua, Zhao Zhi, Li Yanan, Song Zhiguo, Chang Honglei. 2022. Application of digital PCR in the detection of genetically modified plants. Chinese Journal of Biological Control, 38 (5):1143-1148. (in Chinese) doi: 10.16409/j.cnki.2095-039x.2022.09.012
	游英华, 赵志, 李亚楠, 宋治国, 常洪雷. 2022. 数字PCR在转基因植物检测中的应用. 中国生物防治学报, 38 (5):1143-1148. doi: 10.16409/j.cnki.2095-039x.2022.09.012
[76]	Yu N, Han J, Deng T, Chen L, Zhang J, Xing R, Wang P, Zhao G, Chen Y. 2021. A novel analytical droplet digital PCR method for identification and quantification of raw health food material powder from Panax notoginseng. Food Analytical Methods, 14 (3):552-560.
[77]	Zhang J, Luo J, Chen L, Ahmed T, Alotaibi S S, Wang Y, Sun G, Li B, An Q. 2022. Development of droplet digital PCR assay for detection of seed-borne Burkholderia glumae and B. gladioli causing bacterial panicle blight disease of rice. Microorganisms, 10 (6):1223.
[78]	Zmienko A, Samelak-Czajka A, Goralski M, Sobieszczuk-Nowicka E, Kozlowski P, Figlerowicz M. 2015. Selection of reference genes for qPCR- and ddPCR-based analyses of gene expression in senescing barley leaves. PLoS ONE, 10 (2):e0118226.
[79]	Zmienko A, Samelak-Czajka A, Kozlowski P, Szymanska M, Figlerowicz M. 2016. Arabidopsis thaliana population analysis reveals high plasticity of the genomic region spanning MSH2,AT3G18530 and AT3G18535 genes and provides evidence for NAHR-driven recurrent CNV events occurring in this location. BMC Genomics, 17 (1):1-16.

物种 Species	应用方向 Application direction	参考文献 Reference
玉米 Zea mays L.	转基因株系鉴定，基因表达分析，植物病原体诊断 Transgenic lines characterization，gene expression analysis，phytopathogens diagnostics	Corbisier et al.,2010；Xu et al.,2016；Collier et al.,2017；Cottenet et al.,2019；Wang et al.,2023
大豆 Glycine max	植物源性成分检测，转基因株系鉴定 Plant species traceability，transgenic lines characterization	杨硕等,2017；Cottenet et al.,2019；Demeke,2014,2022；
拟南芥 Arabidopsis thaliana	基因表达分析，拷贝数变异，转录本半衰期 Gene expression analysis，copy number variation（CNV）detection，transcript half-life	Notaguchi et al.,2015；Zmienko et al.,2016；Philips et al.,2020
油菜 Brassica napus L.	转基因株系鉴定，拷贝数变异 Transgenic lines characterization，copy number variation（CNV）detection	Demeke,2014,2022；Wu et al.,2017
大麦 Hordeum vulgare L.	基因表达分析，植物病原体诊断，拷贝数变异 Gene expression analysis，phytopathogens diagnostics，copy number variation（CNV）detection	Zmienko et al.,2015；Knight et al.,2022；Gao et al.,2024
甘蔗 Saccharum officinarum	拷贝数变异Copy number variation（CNV）detection	McCord,2016
烟草Nicotiana tabacum	拷贝数变异Copy number variation（CNV）detection	Głowacka et al.,2016
水稻 Oryza sativa L.	拷贝数变异，转基因株系鉴定，植物病原体诊断 Copy number variation（CNV）detection，transgenic lines characterization，phytopathogens diagnostics	Collier et al.,2017；Li et al.,2021；Zhang et al.,2022
马铃薯 Solanum tuberosum	拷贝数变异，转基因株系鉴定，植物病原体诊断 Copy number variation（CNV）detection，transgenic lines characterization，phytopathogens diagnostics	Collier et al.,2017；McCue et al.,2019；Pandey et al.,2020
板栗Castanea mollissima	基因表达分析 Gene expression analysis	Santos et al.,2017
木瓜Pseudocydonia sinensis	拷贝数变异 Copy number variation（CNV）detection	Jia et al.,2017
番茄 Solanum lycopersicum	拷贝数变异，植物病原体诊断 Copy number variation（CNV）detection，phytopathogens diagnostics	Collier et al.,2017；Wang et al.,2022
小麦 Triticum aestivum L.	拷贝数变异，基因表达分析，植物源性成分检测，植物病原体诊断 Copy number variation （CNV）detection，gene expression analysis，plant species traceability，phytopathogens diagnostics	Collier et al.,2017；Li et al.,2020；Morcia et al.,2020；Ren et al.,2022；Wang et al.,2023
柑橘 Citrus reticulata Blanco	拷贝数变异，植物病原体诊断 Copy number variation（CNV） detection，phytopathogens diagnostics	Collier et al.,2017；Keremane et al.,2021
高粱 Sorghum bicolor L.	拷贝数变异 Copy number variation（CNV）detection	Belide et al.,2017
草莓 Fragaria × ananassa	植物病原体诊断，转基因株系鉴定 Phytopathogens diagnostics，transgenic lines characterization	Pérez-López et al.,2017；Duan et al.,2021
棕榈 Trachycarpus fortunei	植物病原体诊断 Phytopathogens diagnostics	Bahder et al.,2018
苜蓿 Medicago sativa L.	转基因株系鉴定 Transgenic lines characterization	Gao et al.,2018
甜瓜 Cucumis melo L.	拷贝数变异 Copy number variation（CNV）detection	Shen et al.,2019b
黄瓜 Cucumis sativus L.	基因表达分析，植物病原体诊断 Gene expression analysis，phytopathogens diagnostics	Shen et al.,2019a；Tian et al.,2023
甜菜 Beta vulgaris L.	基因表达分析，植物病原体诊断 Gene expression analysis，phytopathogens diagnostics	Barone et al.,2019；Chiodi et al.,2021
苋菜 Amaranthus tricolor L.	基因表达分析，拷贝数变异 Gene expression analysis，copy number variation（CNV）detection	Lancíková & Hricová,2020；Yakimowski et al.,2021
苹果 Malus pumila Mill.	植物病原体诊断 Phytopathogens diagnostics	Leichtfried et al.,2020
桃 Prunus persica L.	植物病原体诊断 Phytopathogens diagnostics	Lee et al.,2021
莴苣 Lactuca sativa L.	植物病原体诊断 Phytopathogens diagnostics	Cristiano et al.,2021
葡萄Vitis vinifera L.	基因表达分析，植物源性成分检测，植物病原体诊断 Gene expression analysis，plant species traceability，phytopathogens diagnostics	Morcia et al.,2021；Carrara et al.,2023；Hrycan et al.,2023
三七 Panax notoginseng	植物源性成分检测 Plant species traceability	Yu et al.,2021
猕猴桃 Actinidia chinensis Planch	植物病原体诊断 Phytopathogens diagnostics	Barrett-Manako et al.,2021
白三叶草Trifolium repens	拷贝数变异 Copy number variation（CNV）detection	Narancio et al.,2021
亚麻荠 Camelina sativa	拷贝数变异 Copy number variation（CNV）detection	Darvishi et al.,2021
笋瓜 Cucurbita maxima	植物病原体诊断 Phytopathogens diagnostics	Murolo et al.,2021
蔓越莓 Vaccinium macrocarpon Ait	拷贝数变异 Copy number variation（CNV）detection	Karppinen et al.,2022
梨 Pyrus spp	植物病原体诊断 Phytopathogens diagnostics	Santander et al.,2022
鼠尾草Salvia japonica	基因表达分析 Gene expression analysis	Schmiderer et al.,2023

物种 Species	应用方向 Application direction	参考文献 Reference
玉米 Zea mays L.	转基因株系鉴定，基因表达分析，植物病原体诊断 Transgenic lines characterization，gene expression analysis，phytopathogens diagnostics	Corbisier et al.,2010；Xu et al.,2016；Collier et al.,2017；Cottenet et al.,2019；Wang et al.,2023
大豆 Glycine max	植物源性成分检测，转基因株系鉴定 Plant species traceability，transgenic lines characterization	杨硕等,2017；Cottenet et al.,2019；Demeke,2014,2022；
拟南芥 Arabidopsis thaliana	基因表达分析，拷贝数变异，转录本半衰期 Gene expression analysis，copy number variation（CNV）detection，transcript half-life	Notaguchi et al.,2015；Zmienko et al.,2016；Philips et al.,2020
油菜 Brassica napus L.	转基因株系鉴定，拷贝数变异 Transgenic lines characterization，copy number variation（CNV）detection	Demeke,2014,2022；Wu et al.,2017
大麦 Hordeum vulgare L.	基因表达分析，植物病原体诊断，拷贝数变异 Gene expression analysis，phytopathogens diagnostics，copy number variation（CNV）detection	Zmienko et al.,2015；Knight et al.,2022；Gao et al.,2024
甘蔗 Saccharum officinarum	拷贝数变异Copy number variation（CNV）detection	McCord,2016
烟草Nicotiana tabacum	拷贝数变异Copy number variation（CNV）detection	Głowacka et al.,2016
水稻 Oryza sativa L.	拷贝数变异，转基因株系鉴定，植物病原体诊断 Copy number variation（CNV）detection，transgenic lines characterization，phytopathogens diagnostics	Collier et al.,2017；Li et al.,2021；Zhang et al.,2022
马铃薯 Solanum tuberosum	拷贝数变异，转基因株系鉴定，植物病原体诊断 Copy number variation（CNV）detection，transgenic lines characterization，phytopathogens diagnostics	Collier et al.,2017；McCue et al.,2019；Pandey et al.,2020
板栗Castanea mollissima	基因表达分析 Gene expression analysis	Santos et al.,2017
木瓜Pseudocydonia sinensis	拷贝数变异 Copy number variation（CNV）detection	Jia et al.,2017
番茄 Solanum lycopersicum	拷贝数变异，植物病原体诊断 Copy number variation（CNV）detection，phytopathogens diagnostics	Collier et al.,2017；Wang et al.,2022
小麦 Triticum aestivum L.	拷贝数变异，基因表达分析，植物源性成分检测，植物病原体诊断 Copy number variation （CNV）detection，gene expression analysis，plant species traceability，phytopathogens diagnostics	Collier et al.,2017；Li et al.,2020；Morcia et al.,2020；Ren et al.,2022；Wang et al.,2023
柑橘 Citrus reticulata Blanco	拷贝数变异，植物病原体诊断 Copy number variation（CNV） detection，phytopathogens diagnostics	Collier et al.,2017；Keremane et al.,2021
高粱 Sorghum bicolor L.	拷贝数变异 Copy number variation（CNV）detection	Belide et al.,2017
草莓 Fragaria × ananassa	植物病原体诊断，转基因株系鉴定 Phytopathogens diagnostics，transgenic lines characterization	Pérez-López et al.,2017；Duan et al.,2021
棕榈 Trachycarpus fortunei	植物病原体诊断 Phytopathogens diagnostics	Bahder et al.,2018
苜蓿 Medicago sativa L.	转基因株系鉴定 Transgenic lines characterization	Gao et al.,2018
甜瓜 Cucumis melo L.	拷贝数变异 Copy number variation（CNV）detection	Shen et al.,2019b
黄瓜 Cucumis sativus L.	基因表达分析，植物病原体诊断 Gene expression analysis，phytopathogens diagnostics	Shen et al.,2019a；Tian et al.,2023
甜菜 Beta vulgaris L.	基因表达分析，植物病原体诊断 Gene expression analysis，phytopathogens diagnostics	Barone et al.,2019；Chiodi et al.,2021
苋菜 Amaranthus tricolor L.	基因表达分析，拷贝数变异 Gene expression analysis，copy number variation（CNV）detection	Lancíková & Hricová,2020；Yakimowski et al.,2021
苹果 Malus pumila Mill.	植物病原体诊断 Phytopathogens diagnostics	Leichtfried et al.,2020
桃 Prunus persica L.	植物病原体诊断 Phytopathogens diagnostics	Lee et al.,2021
莴苣 Lactuca sativa L.	植物病原体诊断 Phytopathogens diagnostics	Cristiano et al.,2021
葡萄Vitis vinifera L.	基因表达分析，植物源性成分检测，植物病原体诊断 Gene expression analysis，plant species traceability，phytopathogens diagnostics	Morcia et al.,2021；Carrara et al.,2023；Hrycan et al.,2023
三七 Panax notoginseng	植物源性成分检测 Plant species traceability	Yu et al.,2021
猕猴桃 Actinidia chinensis Planch	植物病原体诊断 Phytopathogens diagnostics	Barrett-Manako et al.,2021
白三叶草Trifolium repens	拷贝数变异 Copy number variation（CNV）detection	Narancio et al.,2021
亚麻荠 Camelina sativa	拷贝数变异 Copy number variation（CNV）detection	Darvishi et al.,2021
笋瓜 Cucurbita maxima	植物病原体诊断 Phytopathogens diagnostics	Murolo et al.,2021
蔓越莓 Vaccinium macrocarpon Ait	拷贝数变异 Copy number variation（CNV）detection	Karppinen et al.,2022
梨 Pyrus spp	植物病原体诊断 Phytopathogens diagnostics	Santander et al.,2022
鼠尾草Salvia japonica	基因表达分析 Gene expression analysis	Schmiderer et al.,2023

Application of Digital PCR in Plant Research

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