Comparison of Endogenous Genes in Maize Based on Digital PCR

doi:10.19586/j.2095-2341.2023.0165

Current Biotechnology ›› 2025, Vol. 15 ›› Issue (1): 78-85.DOI: 10.19586/j.2095-2341.2023.0165

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Comparison of Endogenous Genes in Maize Based on Digital PCR

Xin QI¹(), Xinran LI¹, Yaning GUO², Dan WANG¹, Kai LI¹, Qiong WU³, Liang LI¹()

^1.Institute of Quality Standard and Testing Technology for Agro-Products of Chinese Academy of Agricultural Sciences，Beijing 100081，China
^2.College of Food Science and Bioengineering，Tianjin Agricultural University，Tianjin 300384，China
^3.Beijing Genseed Technology Co. ，Ltd. ，Beijing 100194，China

Received:2023-12-18 Accepted:2024-10-31 Online:2025-01-25 Published:2025-03-07
Contact: Liang LI

基于数字PCR方法的玉米内标准基因比较研究

齐鑫¹(), 李欣然¹, 郭亚宁², 王丹¹, 李凯¹, 吴琼³, 李亮¹()

^1.中国农业科学院农业质量标准与检测技术研究所，北京 100081
^2.天津农学院食品科学与生物工程学院，天津 300384
^3.北京创种科技有限公司，北京 100194

通讯作者: 李亮
作者简介:齐鑫 E-mail: qixin_0908@163.com；
基金资助:
农业生物育种国家科技重大专项(2022ZD04020)

Abstract

Abstract:

With the increasing variety and planting area of transgenic maize， the demand for qualitative and quantitative detection of transgenic maize-related products is increasingly urgent. The selection of stable genetic and specific endogenous genes is of great significance for the accuracy and consistency of transgenic maize identification results. In this study， digital PCR technology was used to screen and compare nine endogenous genes in maize， including hmg， adh1-1， adh1-2， ivrI-1， ivrI-2， zSSIIb-1， zSSIIb-2， zein-1 and zein-2. By optimizing the annealing temperature of the above nine kinds of endogenous genes， the optimal annealing temperature for each gene was found. Taking transgenic maize MON810 as the research object， the above endogenous genes were used to determine the actual samples. The results showed that adh1-1 and adh1-2 could be used for copy number detection of actual samples， which would provide reference for endogenous gene selection and related quantitative detection of transgenic maize in the future， and provid a strong technical support for the management of transgenic crops in China.

Key words: genetically modified maize, GMO detection, endogenous genes, digital PCR

摘要：

随着转基因玉米种类以及种植面积的不断增加，转基因玉米相关产品的定性和定量检测需求日益迫切。选择稳定遗传、特异性强的内标准基因对于转基因玉米鉴定结果的准确性和一致性具有重要意义。利用数字PCR技术对hmg、adh1-1、adh1-2、ivrI-1、ivrI-2、zSSIIb-1、zSSIIb-2、zein-1、zein-2在内的9种玉米内标准基因进行了筛选比较，通过退火温度的优化，找到各自最适的退火温度，并以转基因玉米MON810为研究对象，用上述内标准基因进行实际样品测定。结果表明，adh1-1和adh1-2可以用于实际样品的拷贝数检测。研究结果将为今后转基因玉米内标准基因的选择和相关定量检测提供参考，并对我国转基因作物的管理提供有力的技术支撑。

关键词: 转基因玉米, 转基因检测, 内标准基因, 数字PCR

CLC Number:

Q812

Xin QI, Xinran LI, Yaning GUO, Dan WANG, Kai LI, Qiong WU, Liang LI. Comparison of Endogenous Genes in Maize Based on Digital PCR[J]. Current Biotechnology, 2025, 15(1): 78-85.

齐鑫, 李欣然, 郭亚宁, 王丹, 李凯, 吴琼, 李亮. 基于数字PCR方法的玉米内标准基因比较研究[J]. 生物技术进展, 2025, 15(1): 78-85.

Figures/Tables 9

References 24

1	王凤军,陈强,叶素丹,等.转基因玉米品系及转化体成分四重实时荧光PCR快速鉴定[J].中国粮油学报,2021,36(12):128-135.
	WANG F J, CHEN Q, YE S D, et al.. Rapid identification of transgenic maize lines and transformants by quadruplex real-time PCR[J]. J. Chin. Cereals Oils Assoc., 2021, 36(12): 128-135.
2	TAKABATAKE R, ONISHI M, FUTO S, et al.. Comparison of the specificity,stability,and PCR efficiency of six rice endogenous sequences for detection analyses of genetically modified rice[J]. Food Contr., 2015, 50: 949-955.
3	陈利红,周俊飞,梁晋刚,等.基于二代测序技术的玉米内标准基因扩增子的筛选与评估[J].食品科学,2023,44(20):146-154.
	CHEN L H, ZHOU J F, LIANG J G, et al.. Screening and evaluation of amplicons of maize endogenous reference genes based on next-generation sequencing technology[J]. Food Sci., 2023, 44(20): 146-154.
4	高佳奇,陈硕,王迪等.六种作物内标准基因开发与检测研究进展[J]. 生物技术进展, 2020, 10(6): 613-622.
	GAO J Q, CHEN S, WANG D, et al.. Progress on development and detection of internal reference genes in six crops[J]. Curr. Biotechnol., 2020, 10(6): 613-622.
5	COTTENET G, BLANCPAIN C, CHUAH P F. Performance assessment of digital PCR for the quantification of GM-maize and GM-soya events[J]. Anal. Bioanal. Chem., 2019, 411(11): 2461-2469.
6	DENG T T, HUANG W S, REN J N, et al.. Verification and applicability of endogenous reference genes for quantifying GM rice by digital PCR[J/OL]. Anal. Biochem., 2019, 587: 113442[2024-12-30]. .
7	PATERNÒ A, MARCHESI U, GATTO F, et al.. Finding the joker among the maize endogenous reference genes for genetically modified organism (GMO) detection[J]. J. Agric. Food Chem., 2009, 57(23): 11086-11091.
8	王凤军,杨伊平,叶素丹,等.多重荧光定量PCR快速鉴定转基因玉米品系[J].中国粮油学报,2023,38(4):143-149.
	WANG F J, YANG Y P, YE S D, et al.. Rapid identification of transgenic maize lines by multiplex fluorescence quantitative PCR[J]. J. Chin. Cereals Oils Assoc., 2023, 38(4): 143-149.
9	李凌燕,张旭冬,陈子言,等.转基因抗虫耐除草剂玉米MON87411精准定量检测方法的建立[J].生物安全学报,2023,32(1):38-45.
	LI L Y, ZHANG X D, CHEN Z Y, et al.. Establishment of accurate quantitative detection method for insect-resistant and herbicide-tolerant maize MON87411[J]. J. Biosaf., 2023, 32(1): 38-45.
10	周圆,单长林,李孝军,等.运用微滴式数字PCR技术检测转基因玉米品系的研究[J].安徽农业科学,2018,46(14):175-178.
	ZHOU Y, SHAN C L, LI X J, et al.. Study on detection of genetically modified maize lines using microdrop digital PCR technique[J]. J. Anhui Agric. Sci., 2018, 46(14): 175-178.
11	FOLLONI S, BELLOCCHI G, PROSPERO A, et al.. Statistical evaluation of real-time PCR protocols applied to quantify genetically modified maize[J]. Food Anal. Meth., 2010, 3(4): 304-312.
12	WEI S, WANG C G, ZHU P Y, et al.. A high-throughput multiplex tandem PCR assay for the screening of genetically modified maize[J]. LWT, 2018, 87: 169-176.
13	PAPAZOVA N, ZHANG D, GRUDEN K, et al.. Evaluation of the reliability of maize reference assays for GMO quantification[J]. Anal. Bioanal. Chem., 2010, 396(6): 2189-2201.
14	肖芳,李俊,王颢潜等.转基因玉米NK603转化体/zSSIIb内标基因二重微滴数字PCR方法的建立及应用[J]. 中国农业科学, 2021, 54(22): 4728-4739.
	XIAO F, LI J, WANG H Q, et al.. Establishment and application of A duplex ddPCR method to quantify the NK603/zSSIIb copy number ratio in transgenic maize NK603[J]. Sci. Agric. Sin., 2021, 54(22): 4728-4739.
15	SCHOLDBERG T A, NORDEN T D, NELSON D D, et al.. Evaluating precision and accuracy when quantifying different endogenous control reference genes in maize using real-time PCR[J]. J. Agric. Food Chem., 2009, 57(7): 2903-2911.
16	DMIQE G, HUGGETT J F.The digital MIQE guidelines update:minimum information for publication of quantitative digital PCR experiments for 2020[J]. Clin. Chem., 2020, 66(8): 1012-1029.
17	BASU A S. Digital assays part I: partitioning statistics and digital PCR[J]. SLAS Technol., 2017, 22(4): 369-386.
18	CORBISIER P, BUTTINGER G, SAVINI C, et al.. Expression of GM content in mass fraction from digital PCR data[J/OL]. Food Contr., 2022, 133(Pt B): 108626[2024-12-30]. .
19	ROWLANDS V, RUTKOWSKI A J, MEUSER E, et al.. Optimisation of robust singleplex and multiplex droplet digital PCR assays for high confidence mutation detection in circulating tumour DNA[J/OL]. Sci. Rep., 2019, 9: 12620[2024-12-30]. .
20	ZHANG H, LAŠŠÁKOVÁ S, YAN Z, et al.. Digital polymerase chain reaction duplexing method in a single fluorescence channel[J/OL]. Anal. Chim. Acta., 2023, 1238: 340243[2024-12-30]. .
21	DEBSKI P R, GARSTECKI P. Designing and interpretation of digital assays: concentration of target in the sample and in the source of sample[J]. Biomol. Detect. Quantif., 2016, 10: 24-30.
22	国家质量监督检验检疫总局,中国国家标准化管理委员会. 水溶液中核酸的浓度和纯度检测紫外分光光度法: [S].北京:中国标准出版社,2017.
23	刘晓,朱鹏宇,景小艳,等.双重数字PCR在转基因大豆检测中的应用[J].生物技术进展,2020,10(1):60-66.
	LIU X, ZHU P Y, JING X Y, et al.. Application of duplex droplet digital PCR for detection of genetically modified soybean[J]. Curr. Biotechnol., 2020, 10(1): 60-66.
24	卢海强,焦新雅,吴思源,等.数字PCR在食源性致病菌检测中的应用进展[J]. 生物技术进展, 2021, 11(3): 260-268.
	LU H Q, JIAO X Y, WU S Y, et al.. Application progress on the digital PCR in detection of foodborne pathogenic bacteria[J]. Curr. Biotechnol., 2021, 11(3): 260-268.

基因名称		引物探针序列（5'→3'）	扩增子/bp	参考文献
hmg	F-1	5′-TTGGACTAGAAATCTCGTGCTGA-3′	79	[7,11,15]
	R-1	5′-GCTACATAGGGAGCCTTGTCCT-3′
	P-1	5′-CAATCCACACAAACGCACGCGTA-3′
adh1-1	F-1	5′-CGTCGTTTCCCATCTCTTCCTCC-3′	135	[14-15]
	R-1	5′-CCACTCCGAGACCCTCAGTC-3′
	P-1	5′-AATCAGGGCTCATTTTCTCGCTCCTCA-3′
adh1-2	F-2	5′-CCTTCTTGGCGGCTTATCTG-3′	70	[7，13]
	R-2	5′-CCAGCCTCATGGCCAAAG-3′
	P-2	5′-CTTAGGGGCAGACTCCCGTGTTCCCT-3′
ivr1-1	F-1	5′-CGCTCTGTACAAGCGTGC-3′	104	[15]
	R-1	5′-GCAAAGTGTTGTGCTTGGACC-3′
	P-1	5′-CACGTGAGAATTTCCGTCTACTCGAGCCT-3′
ivr1-2	F-2	5′-TGGCGGACGACGACTTGT-3′	79	[4]
	R-2	5′-AAAGTTTGGAGGCTGCCGT-3′
	P-2	5′-CGAGCAGACCGCCGTGTACTTCTACC-3′
zein-1	F-1	5′-CGTGTCCGTCCCTGATGC-3′	83	[4]
	R-1	5′-AGGCGTCATCATCTGTGGC-3′
	P-1	5′-CAACTGTTGGCCTTACCGCTTCAGACG-3′
zein-2	F-2	5′-GCCATTGGGTACCATGAACC-3′	104	[13]
	R-2	5′-AGGCCAACAGTTGCTGCAG-3′
	P-2	5′-AGCTTGATGGCGTGTCCGTCCCT-3′
zSSIIb-1	F-1	5′-CTCCCAATCCTTTGACATCTGC-3′	151	[15]
	R-1	5′-TCGATTTCTCTCTTGGTGACAGG-3′
	P-1	5′-AGCAAAGTCAGAGCGCTGCAATGCA-3′
zSSIIb-2	F-2	5′-CCAATCCTTTGACATCTGCTCC-3′	114	[15]
	R-2	5′-GATCAGCTTTGGGTCCGGA-3′
	P-2	5′-AGCAAAGTCAGAGCGCTGCAATGCA-3′

基因名称		引物探针序列（5'→3'）	扩增子/bp	参考文献
hmg	F-1	5′-TTGGACTAGAAATCTCGTGCTGA-3′	79	[7,11,15]
	R-1	5′-GCTACATAGGGAGCCTTGTCCT-3′
	P-1	5′-CAATCCACACAAACGCACGCGTA-3′
adh1-1	F-1	5′-CGTCGTTTCCCATCTCTTCCTCC-3′	135	[14-15]
	R-1	5′-CCACTCCGAGACCCTCAGTC-3′
	P-1	5′-AATCAGGGCTCATTTTCTCGCTCCTCA-3′
adh1-2	F-2	5′-CCTTCTTGGCGGCTTATCTG-3′	70	[7，13]
	R-2	5′-CCAGCCTCATGGCCAAAG-3′
	P-2	5′-CTTAGGGGCAGACTCCCGTGTTCCCT-3′
ivr1-1	F-1	5′-CGCTCTGTACAAGCGTGC-3′	104	[15]
	R-1	5′-GCAAAGTGTTGTGCTTGGACC-3′
	P-1	5′-CACGTGAGAATTTCCGTCTACTCGAGCCT-3′
ivr1-2	F-2	5′-TGGCGGACGACGACTTGT-3′	79	[4]
	R-2	5′-AAAGTTTGGAGGCTGCCGT-3′
	P-2	5′-CGAGCAGACCGCCGTGTACTTCTACC-3′
zein-1	F-1	5′-CGTGTCCGTCCCTGATGC-3′	83	[4]
	R-1	5′-AGGCGTCATCATCTGTGGC-3′
	P-1	5′-CAACTGTTGGCCTTACCGCTTCAGACG-3′
zein-2	F-2	5′-GCCATTGGGTACCATGAACC-3′	104	[13]
	R-2	5′-AGGCCAACAGTTGCTGCAG-3′
	P-2	5′-AGCTTGATGGCGTGTCCGTCCCT-3′
zSSIIb-1	F-1	5′-CTCCCAATCCTTTGACATCTGC-3′	151	[15]
	R-1	5′-TCGATTTCTCTCTTGGTGACAGG-3′
	P-1	5′-AGCAAAGTCAGAGCGCTGCAATGCA-3′
zSSIIb-2	F-2	5′-CCAATCCTTTGACATCTGCTCC-3′	114	[15]
	R-2	5′-GATCAGCTTTGGGTCCGGA-3′
	P-2	5′-AGCAAAGTCAGAGCGCTGCAATGCA-3′

基因名称	59.0 ℃	58.6 ℃	58.0 ℃	57.1 ℃	55.9 ℃	55.0 ℃	平均值±标准差	RSD/%
hmg	3 550.0	3 643.3	3 690.0	3 653.3	3 660.0	3 736.7	3 655.6±61.78	1.69
adh1-1	2 630.0	2 663.3	2 676.7	2 676.7	2 640.0	2 673.3	2 660.0±20.22	0.76
adh1-2	3 190.0	3 230.0	3 216.7	3 260.0	3 236.7	3 200.0	3 222.2±25.53	0.79
ivr1-1	2 733.3	2 753.3	2 806.7	2 763.3	2 813.3	2 793.3	2 777.2±32.00	1.15
ivr1-2	3 600.0	3 596.7	3 790.0	3 790.0	3 700.0	3 716.7	3 698.9±86.20	2.33
zein-1	2 613.3	2 666.7	2 763.3	3 030.0	3 583.3	4 083.3	3 123.3±589.94	18.89
zein-2	2 823.3	2 833.3	2 923.3	2 900.0	2 926.7	2 900.0	2 884.4±45.00	1.56
zSSIIb-1	3 443.3	3 416.7	3 493.3	3 496.7	3 476.7	3 510.0	3 472.8±35.80	1.03
zSSIIb-2	4 070.0	4 123.3	4 136.7	4 186.7	4 180.0	4 040.0	4 122.8±58.63	1.42

基因名称	59.0 ℃	58.6 ℃	58.0 ℃	57.1 ℃	55.9 ℃	55.0 ℃	平均值±标准差	RSD/%
hmg	3 550.0	3 643.3	3 690.0	3 653.3	3 660.0	3 736.7	3 655.6±61.78	1.69
adh1-1	2 630.0	2 663.3	2 676.7	2 676.7	2 640.0	2 673.3	2 660.0±20.22	0.76
adh1-2	3 190.0	3 230.0	3 216.7	3 260.0	3 236.7	3 200.0	3 222.2±25.53	0.79
ivr1-1	2 733.3	2 753.3	2 806.7	2 763.3	2 813.3	2 793.3	2 777.2±32.00	1.15
ivr1-2	3 600.0	3 596.7	3 790.0	3 790.0	3 700.0	3 716.7	3 698.9±86.20	2.33
zein-1	2 613.3	2 666.7	2 763.3	3 030.0	3 583.3	4 083.3	3 123.3±589.94	18.89
zein-2	2 823.3	2 833.3	2 923.3	2 900.0	2 926.7	2 900.0	2 884.4±45.00	1.56
zSSIIb-1	3 443.3	3 416.7	3 493.3	3 496.7	3 476.7	3 510.0	3 472.8±35.80	1.03
zSSIIb-2	4 070.0	4 123.3	4 136.7	4 186.7	4 180.0	4 040.0	4 122.8±58.63	1.42

MON810稀释倍数	拷贝数·µL^-1				均值	RSD/%
MON810稀释倍数	重复1	重复2	重复3	重复4	均值	RSD/%
50 000	5 470	5 680	5 620	5 840	5 652.5	2.71
70 000	3 680	3 760	3 600	3 880	3 730.0	3.20
90 000	3 350	3 380	3 210	3 360	3 325.0	2.34

Comparison of Endogenous Genes in Maize Based on Digital PCR

基于数字PCR方法的玉米内标准基因比较研究

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靶标	拷贝数·µL^-1				均值	RSD/%
靶标	重复1	重复2	重复3	重复4	均值	RSD/%
hmg	3 540	3 860	3 910	3 610	3 730.0	4.89
adh1-1	3 340	3 410	3 340	3 430	3 380.0	1.39
adh1-2	3 920	4 030	4 000	3 970	3 980.0	1.18
ivr1-1	3 260	3 400	3 440	3 420	3 380.0	2.42
ivr1-2	No Call	No Call	No Call	No Call	-	-
zSSIIb-1	3 060	3 160	3 180	3 280	3 170.0	2.84
zSSIIb-2	3 380	3 400	3 450	3 380	3 402.5	0.97
zein-1	4 100	4 120	4 130	4 110	4 115.0	0.31
zein-2	4 120	4 130	4 120	4 040	4 102.5	1.02

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靶标	拷贝数·µL^-1				均值	标准差	RSD/%
靶标	重复1	重复2	重复3	重复4	均值	标准差	RSD/%
MON810-5	547	568	562	584	565.25	15.31	0.03
adh1-1	334	341	334	343	338.00	4.69	0.01
外源/内标	164%	167%	168%	170%	167%	0.03	0.02
MON810-7	368	376	360	388	373.00	11.94	0.03
adh1-1	334	341	334	343	338.00	4.69	0.01
外源/内标	110%	110%	108%	113%	110%	0.02	0.02
MON810-9	335	338	321	336	332.50	7.77	0.02
adh1-1	334	341	334	343	338.00	4.69	0.01
外源/内标	100%	99%	96%	98%	98%	0.02	0.02

靶标	拷贝数·µL^-1				均值	标准差	RSD/%
靶标	重复1	重复2	重复3	重复4	均值	标准差	RSD/%
MON810-5	547	568	562	584	565.25	15.31	0.03
adh1-1	334	341	334	343	338.00	4.69	0.01
外源/内标	164%	167%	168%	170%	167%	0.03	0.02
MON810-7	368	376	360	388	373.00	11.94	0.03
adh1-1	334	341	334	343	338.00	4.69	0.01
外源/内标	110%	110%	108%	113%	110%	0.02	0.02
MON810-9	335	338	321	336	332.50	7.77	0.02
adh1-1	334	341	334	343	338.00	4.69	0.01
外源/内标	100%	99%	96%	98%	98%	0.02	0.02

靶标	拷贝·µL^-1				均值	标准差	RSD/%
靶标	重复1	重复2	重复3	重复4	均值	标准差	RSD/%
MON810-5	547	568	562	584	565.25	15.31	0.03
adh1-2	392	403	400	397	398.00	4.69	0.01
外源/内标	140%	141%	141%	147%	142%	0.03	0.02
MON810-7	368	376	360	388	373.00	11.94	0.03
adh1-2	392	403	400	397	398.00	4.69	0.01
外源/内标	94%	93%	90%	98%	94%	0.03	0.03
MON810-9	335	338	321	336	332.50	7.77	0.02
adh1-2	392	403	400	397	398.00	4.69	0.01
外源/内标	85%	84%	80%	85%	84%	0.02	0.03

靶标	拷贝·µL^-1				均值	标准差	RSD/%
靶标	重复1	重复2	重复3	重复4	均值	标准差	RSD/%
MON810-5	547	568	562	584	565.25	15.31	0.03
adh1-2	392	403	400	397	398.00	4.69	0.01
外源/内标	140%	141%	141%	147%	142%	0.03	0.02
MON810-7	368	376	360	388	373.00	11.94	0.03
adh1-2	392	403	400	397	398.00	4.69	0.01
外源/内标	94%	93%	90%	98%	94%	0.03	0.03
MON810-9	335	338	321	336	332.50	7.77	0.02
adh1-2	392	403	400	397	398.00	4.69	0.01
外源/内标	85%	84%	80%	85%	84%	0.02	0.03