数字PCR技术在植物研究中的应用

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

摘要/Abstract

摘要：

数字PCR技术是近年来兴起的一种高灵敏核酸绝对定量分析技术，无需标准品和绘制标准曲线即可实现对样品核酸初始浓度的绝对定量，具有高灵敏度、高特异性和高精确度的特点。本文阐述了数字PCR技术的基本原理，综述了数字PCR技术在转基因植物检测、植物病原体诊断、基因拷贝数变异和基因表达研究等方面的应用，并对其在植物中的应用前景进行展望。

关键词: 数字PCR, 植物研究, 转基因植物, 植物病原体诊断, 绝对定量

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

高媛, 马月花, 张盼盼, 束胜, 陈素梅. 数字PCR技术在植物研究中的应用[J]. 园艺学报, 2024, 51(12): 2977-2988.

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.

图/表 2

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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