GMO molecular characterization details

GMO detailed information

GMO:
DP23211
Alias:
UID:
DP-Ø23211-2
Tradename:
Companies/Institutions:
Pioneer Hi-Bred International
Species:
Zea mays
(maize, corn)
Traits:
EU authorisation:

General Information

Description:

The maize event DP23211 was produced by Agrobacterium-mediated SSI (site-specific integration) transformation with the vector PHP74643. The event contains four gene cassettes:

The first gene cassette contains an siRNA system targeting the smooth septate junction protein 1 gene (SSJ1) in Diabrotica virgifera and hereby conferring insect resistance.
The second cassette contains the gene ipd072Aa from Pseudomonas chlororaphis. The protein IPD072Aa confers protection against certain coleopteran pests.
The third cassette contains the phosphinothricin acetyl transferase gene (pat) from Streptomyces viridochromogenes. The PAT enzyme confers tolerance to phosphinothricin (a.k.a. glufosinate).
The fourth gene cassette contains the phosphomannose isomerase (pmi) gene from Escherichia coli. Expression of the PMI protein in plants servers as a selectable marker which allows plant tissue growth with mannose as the carbon source.

Southern by Sequencing (SbS) analysis of DP23211 demonstrated that there is a single, intact insertion. Furthermore, the sequence of the insert and the flanking borders was obtained by Sanger sequencing.

Genetic and phenotypic stability of the inserts in DP23211 was confirmed by molecular and segregation analysis of different DP23211 maize generations.

Links regarding to this GMO:

MAFF Japan http://www.affrc.maff.go.jp/attach/pdf/20190201-12.pdf,

WIPO https://patentimages.storage.googleapis.com/ee/ff/f8/90f1f2112adabe/WO2019209700A1.pdf,

BCH https://bch.cbd.int/en/database/record?documentID=116060,

USDA-APHIS https://www.aphis.usda.gov/brs/aphisdocs/20-20301p.pdf,

EFSA https://open.efsa.europa.eu/study-inventory/EFSA-Q-2019-00807

Transformation / Modification technique:

Agrobacterium-induced

Inserts / Modification

Name:	insert_DP23211
Comment:	The 71116 bp vector PHP74643 has been used for the creation of event DP23211. Only a partial sequence (between FRT1 and FRT87) comprising of 4 gene cassettes has been inserted into the genome of the cultivar.
Map:

Genetic Elements:

Reference: patent documents

Elements:

Order	Name	Donor	Position	Definition	Trait	Comment
1	V-intervening-SYNTH	Synthetic	-			flippase recombination target site (FRT1)
2	CS-pmi-ECOLX	Escherichia coli	-	Phosphomannose isomerase. Reed et al (2001). In Vitro Cell Dev Biol Plant, 37, 127-132	Mannose metabolism
3	T-pinII-SOLTU	Solanum tuberosum	-	3' untranslated region (3' UTR) of the proteinase inhibitor II gene; 0.31kb terminator sequence.
4	T-z19-MAIZE	Zea mays	-	Terminator of the 19-kDA zein gene from Zea mays. GenBank KX247647
5	P-act1-ORYSA	Oryza sativa	-	Promoter of the actin 1 gene. McElroy et al. (1990). The Plant Cell, 2, 163-171
6	I-1_act1-ORYSA	Oryza sativa	-
7	CS-pat-STRVR	Streptomyces viridochromogenes	-	Phosphinothricin acetyltransferase gene derived from Streptomyces viridochromogenes a.k.a. pat gene; shares 85 per cent homology at the amino acid level with the bar gene. Wohlleben et al. (1988) Gene 70:25-37	Glufosinate tolerance	maize optimized phosphinothricin acetyltransferase gene (mo-pat)
8	T-35S-CaMV	Cauliflower mosaic virus	-	3' untranslated region (3' UTR) of the 35S gene from Cauliflower mosaic virus. Gardner et al. (1981). Nucleic Acids Res, 9:2871-2888
9	T-ubi-SORBI	Sorghum bicolor	-	Terminator of the ubiquitin gene. Phytozome gene ID Sobic.004G049900.l
10	T-gkaf-SORBI	Sorghum bicolor	-	Terminator of the gamma-kafirin gene; De Freitas et al. (1994). Molecular and General Genetics, 245:177-186
11	P-ubi1-MAIZE	Zea mays	-	Promoter region of the polyubiquitin gene. Christensen et al. (1992). Plant Mol Biol, 18, 675-689		length: 900 bp
12	I-ubi1-MAIZE	Zea mays	-	5' UTR of noncoding region of ubiquitin gene 1.		length: 83 bp; 5‘ UTR
13	I-1_ubi1-MAIZE	Zea mays	-			length: 1013 bp
14	CS-SSJ1 _RNAi_u-DIAVI	Diabrotica virgifera virgifera	-	Fragment of the smooth septate junction protein 1 gene (SSJ1) designed to match that from D. virgifera (Western corn rootworm).	Coleoptera resistance	length: 210 bp
15	I-1_adh1-MAIZE	Zea mays	-			length: 100 bp; connector sequence derived from the Intron 1 region of the Zea mays alcohol dehydrogenase gene, containing the first 50 bp of the 5‘ end and the last 50 bp of the 3‘ end
16	CS-SSJ1 _RNAi_u-DIAVI	Diabrotica virgifera virgifera	-	Fragment of the smooth septate junction protein 1 gene (SSJ1) designed to match that from D. virgifera (Western corn rootworm).	Coleoptera resistance	length: 210 bp; complementary to element 14
17	T-gz27-MAIZE	Zea mays	-	Terminator of the 27-kDA gamma zein gene.		length: 480 bp
18	T-ubi14-ARATH	Arabidopsis thaliana	-	3' untranslated region (3' UTR) of the ubiquitin 14 gene. Callis et al. (1995). Genetics 139: 921-939		length: 902 bp
19	T-In2_1-MAIZE	Zea mays	-	Terminator region from the Zea mays In2-1 gene. Hershey and Stoner (1991), Plant Mol Biol 17(4):679-90. doi: 10.1007/BF00037053.		length: 494 bp
20	P-BSV(YV)	Banana streak virus	-	Promoter region from the Banana streak virus (Acuminata Yunnan strain); Schenk et al. (2001). Plant Mol Biol, 47(3):399-412.		length: 414 bp
21	I-unknown protein-MAIZE	Zea mays	-	Intron region from the Zea mays ortholog of an Oryza sativa hypothetical protein.		length: 856 bp
22	CS-ipd072Aa-PSECL	Pseudomonas chlororaphis	-	Insecticidal protein ipd072Aa gene.	Coleoptera resistance	length: 261 bp
23	T-JR1_putative-ARATH	Arabidopsis thaliana	-	Terminator region of putative gene of the mannose-binding lectin superfamily protein (JR1).		length: 573 bp
24	V-intervening sequences		-			length: 48 bp; FRT87, Modified Flippase recombination target site derived from Sachhammyces cenvistae

Related DNA Sequences:

Acc.No. Reference Sequence Size (bp) Annotation

patent documents No

Transformation Vectors

Name:	vector_PHP74643
Comment:	The 71116 bp vector PHP74643 has been used for the creation of event DP23211 and DP62151.
Map:

Genetic Elements:

Reference: patent documents

Elements:

Order	Name	Donor	Size (bp)	Position	Definition	Trait	Comment
1	V-RB-RHIRD	Agrobacterium tumefaciens		-
2	P-nopaline synthase			-
3	CS-wus2_MAIZE	Zea mays		-	Homeobox-transcription factor 122 (Putative homeobox DNA-binding domain superfamily protein) from Zea mays. Mayer KFX et al.(1998).Cell, 95: 805-815; Lowe K et al. (2016). Plant Cell, 28:1998-2015	Alteration in growth, development or product quality
4	T-pinII-SOLTU	Solanum tuberosum		-	3' untranslated region (3' UTR) of the proteinase inhibitor II gene; 0.31kb terminator sequence.
5	P-ubi1-MAIZE	Zea mays		-	Promoter region of the polyubiquitin gene. Christensen et al. (1992). Plant Mol Biol, 18, 675-689
6	I-1_ubi1-MAIZE	Zea mays		-			5' UTR
7	I-1_ubi1-MAIZE	Zea mays		-
8	CS-odp2_MAIZE	Zea mays		-	odp2 is an ovule development protein from Zea mays, GenBank XM008676474, Lowe K et al. (2016). Plant Cell, 28:1998-2015	Alteration in growth, development or product quality
9	T-pinII-SOLTU	Solanum tuberosum		-	3' untranslated region (3' UTR) of the proteinase inhibitor II gene; 0.31kb terminator sequence.
10	T-z19-MAIZE	Zea mays		-	Terminator of the 19-kDA zein gene from Zea mays. GenBank KX247647
11	P-ubi1-MAIZE	Zea mays		-	Promoter region of the polyubiquitin gene. Christensen et al. (1992). Plant Mol Biol, 18, 675-689
12	I-1_ubi1-MAIZE	Zea mays		-			5' UTR
13	I-1_ubi1-MAIZE	Zea mays		-
14	R-flp1_YEAST	Saccharomyces cerevisiae		-	Exon of the FLP recombinase from S. cerevisiae. The gene encodes the protein 1 region. Sadowski (1995). Prog Nucleic Acid Res Mol Biol, 51:53-91		maize codon optimized-flp Exon1
15	I-2_psbr-SOLTU	Solanum tuberosum		-	Intron 2 of the psbr gene (also known as light-inducible tissue-specific LS1 gene); length: 193bp.		st-LS1 Intron
16	R-flp2_YEAST	Saccharomyces cerevisiae		-	Exon of the FLP recombinase from S. cerevisiae. The gene encodes the protein 2 region. Sadowski (1995). Prog Nucleic Acid Res Mol Biol, 51:53-91		maize codon optimized-flp Exon2
17	T-pinII-SOLTU	Solanum tuberosum		-	3' untranslated region (3' UTR) of the proteinase inhibitor II gene; 0.31kb terminator sequence.
18	E-35S-CaMV	Cauliflower mosaic virus		-
19	P-ltp2-HORVU	Hordeum vulgare		-	Promoter of a lipid transfer protein gene. Kalla (1994). The Plant Joumal, 6(6):849-860
20	CS-DsRed2-SYNTH	Synthetic		-	Variant with one alteration (Alt1) of Clontech's DsRed protein, isolated from a marine coral, Discosoma sp., with unchanged amino acid sequence	DsRed2 activity
21	T-35S-CaMV	Cauliflower mosaic virus		-	3' untranslated region (3' UTR) of the 35S gene from Cauliflower mosaic virus. Gardner et al. (1981). Nucleic Acids Res, 9:2871-2888
22	T-35S-CaMV	Cauliflower mosaic virus		-	3' untranslated region (3' UTR) of the 35S gene from Cauliflower mosaic virus. Gardner et al. (1981). Nucleic Acids Res, 9:2871-2888
23	V-intervening-SYNTH	Synthetic		-			flippase recombination target site (FRT1)
24	CS-pmi-ECOLX	Escherichia coli		-	Phosphomannose isomerase. Reed et al (2001). In Vitro Cell Dev Biol Plant, 37, 127-132	Mannose metabolism
25	T-pinII-SOLTU	Solanum tuberosum		-	3' untranslated region (3' UTR) of the proteinase inhibitor II gene; 0.31kb terminator sequence.
26	T-z19-MAIZE	Zea mays		-	Terminator of the 19-kDA zein gene from Zea mays. GenBank KX247647
27	P-act1-ORYSA	Oryza sativa		-	Promoter of the actin 1 gene. McElroy et al. (1990). The Plant Cell, 2, 163-171
28	I-1_act1-ORYSA	Oryza sativa		-
29	CS-pat-STRVR	Streptomyces viridochromogenes		-	Phosphinothricin acetyltransferase gene derived from Streptomyces viridochromogenes a.k.a. pat gene; shares 85 per cent homology at the amino acid level with the bar gene. Wohlleben et al. (1988) Gene 70:25-37	Glufosinate tolerance	maize codon optimized (mo-pat)
30	T-35S-CaMV	Cauliflower mosaic virus		-	3' untranslated region (3' UTR) of the 35S gene from Cauliflower mosaic virus. Gardner et al. (1981). Nucleic Acids Res, 9:2871-2888
31	T-ubi-SORBI	Sorghum bicolor		-	Terminator of the ubiquitin gene. Phytozome gene ID Sobic.004G049900.l
32	T-gkaf-SORBI	Sorghum bicolor		-	Terminator of the gamma-kafirin gene; De Freitas et al. (1994). Molecular and General Genetics, 245:177-186
33	V-MCS-SYNTH	Synthetic	87	20691-20777
34	P-ubi1-MAIZE	Zea mays	900	20778-21677	Promoter region of the polyubiquitin gene. Christensen et al. (1992). Plant Mol Biol, 18, 675-689
35	I-ubi1-MAIZE	Zea mays	83	21678-21760	5' UTR of noncoding region of ubiquitin gene 1.		5‘ UTR
36	I-1_ubi1-MAIZE	Zea mays	1013	21761-22773
37	V-MCS-SYNTH	Synthetic	39	22774-22812
38	CS-SSJ1 _RNAi_u-DIAVI	Diabrotica virgifera virgifera	210	22813-23022	Fragment of the smooth septate junction protein 1 gene (SSJ1) designed to match that from D. virgifera (Western corn rootworm).	Coleoptera resistance
39	V-MCS-SYNTH	Synthetic	21	23023-23043
40	I-1_adh1-MAIZE	Zea mays	100	23044-23143			Connector sequence derived from the Intron 1 region of the Zea Mays alcohol dehydrogenase gene, containing the first 50 bp of the 5‘ end and the last 50 bp of the 3‘ end
41	V-MCS-SYNTH	Synthetic	21	23144-23164
42	CS-SSJ1 _RNAi_u-DIAVI	Diabrotica virgifera virgifera	210	23165-23374	Fragment of the smooth septate junction protein 1 gene (SSJ1) designed to match that from D. virgifera (Western corn rootworm).	Coleoptera resistance	complementary to element 38
43	V-MCS-SYNTH	Synthetic	34	23375-23408
44	T-gz27-MAIZE	Zea mays	480	23409-23888	Terminator of the 27-kDA gamma zein gene.
45	V-MCS-SYNTH	Synthetic	6	23889-23894
46	T-ubi14-ARATH	Arabidopsis thaliana	902	23895-24796	3' untranslated region (3' UTR) of the ubiquitin 14 gene. Callis et al. (1995). Genetics 139: 921-939
47	V-MCS-SYNTH	Synthetic	6	24797-24802
48	T-In2_1-MAIZE	Zea mays	494	24803-25296	Terminator region from the Zea mays In2-1 gene. Hershey and Stoner (1991), Plant Mol Biol 17(4):679-90. doi: 10.1007/BF00037053.
49	V-MCS-SYNTH	Synthetic	57	25297-25353
50	V-intervening sequences		24	25354-25377			attB2, Bacteriophage lambda integrase recombination site from the Invitrogen Gateway ® cloning system
51	V-MCS-SYNTH	Synthetic	37	25378-25414
52	P-BSV(YV)	Banana streak virus	414	25415-25828	Promoter region from the Banana streak virus (Acuminata Yunnan strain); Schenk et al. (2001). Plant Mol Biol, 47(3):399-412.
53	V-MCS-SYNTH	Synthetic	19	25829-25847
54	I-unknown protein-MAIZE	Zea mays	856	25848-26703	Intron region from the Zea mays ortholog of an Oryza sativa hypothetical protein.
55	V-MCS-SYNTH	Synthetic	9	26704-26712
56	CS-ipd072Aa-PSECL	Pseudomonas chlororaphis	261	26713-26973	Insecticidal protein ipd072Aa gene.	Coleoptera resistance
57	V-MCS-SYNTH	Synthetic	6	26974-26979
58	T-JR1_putative-ARATH	Arabidopsis thaliana	573	26980-27552	Terminator region of putative gene of the mannose-binding lectin superfamily protein (JR1).
59	V-MCS-SYNTH	Synthetic	39	27553-27591
60	V-intervening sequences		21	27592-27612			attB3, Bacteriophage lambda integrase recombination site from the Invitrogen Gateway ® cloning system
61	V-MCS-SYNTH	Synthetic	121	27613-27733
62	V-intervening sequences		48	27734-27781			FRT87, Modified Flippase recombination target site derived from Sachhammyces cenvistae

Related DNA Sequences

Acc.No.	Reference	Sequence	Size (bp)	Annotation
	patent documents	No

Detection Methods

18 results

Name	Type	Target	Description	Validation	Standardisation	Verification
QL-ELE-AINT 2-5'/AINT 2-3'	element-specific	I-actin	Qualitative real-time PCR (TaqMan) method for detection of rice actin 1 intron (I-ractI) (Mano et al. 2009).	in-house validation	none	1
QL-ELE-Patf-n2/Patr-n2	element-specific	CS-pat-STRVR	Qualitative real-time PCR (TaqMan) method for detection of the pat gene (Xu et al., 2006)	in-house validation	none	1
QL-ELE-P-Rice actin-F/P-Rice actin-R	element-specific	P-act1-ORYSA	Qualitative real-time PCR (TaqMan) method for detection of rice actin 1 promoter (P-ractI) (Debode et al. 2013).	in-house validation	none	1
QL-ELE-P-Ubi-F/P-Ubi-R	element-specific	P-ubi1-MAIZE	Qualitative real-time PCR (TaqMan) method for detection of ubiquitin promoter of Zea mays (Debode et al., 2013)	in-house validation	none	1
QL-ELE-T-35S-F/T-35S-R	element-specific	T-Cauliflower mosaic virus	Qualitative real-time PCR (TaqMan) method for detection of the 35S terminator from Cauliflower mosaic virus (Debode et al., 2013)	in-house validation	none	1
QL-TAX-ZM-002	taxon-specific	Zea mays	Qualitative real-time PCR (SYBRGreen) method for detection of maize alcoholdeydrogenase 1 gene (Barbau-Piednoir et al., 2014)	ring trial validation	unknown	1
QL-TAX-ZM-003	taxon-specific	Zea mays	Qualitative conventional PCR method for detection of the maize invertase gene (ISO 21569)	ring trial validation	ISO standard	1
QT-EVE-ZM-035	event-specific	DP23211	Quantitative real-time PCR (TaqMan) method for detection of maize event DP23211 (EU-RL GMFF GMOMETHODS database as of February 15, 2024)	ring trial validation	EU reference method	1
QT-TAX-ZM-001	taxon-specific	Zea mays	Quantitative real-time PCR (TaqMan) method for detection of maize alcohol dehydrogenase 1 (adh1) gene. Estimated number of target copies per haploid genome (Jacchia et al. 2018): 1	ring trial validation	ISO standard	1
QT-TAX-ZM-002	taxon-specific	Zea mays	Quantitative real-time PCR (TaqMan) method for detection of the maize high-mobility-group A (hmgA) gene (ISO 21570). Estimated number of target copies per haploid genome (Jacchia et al. 2018): 1	ring trial validation	ISO standard	1
QT-TAX-ZM-003	taxon-specific	Zea mays	Quantitative real-time PCR (TaqMan) method for detection of maize alcohol dehydrogenase 1 (adh1) gene (EU-RL GMFF GMOMETHODS database as of October 01, 2014)	ring trial validation	EU reference method	1
QT-TAX-ZM-004	taxon-specific	Zea mays	Quantitative real-time PCR (TaqMan) for detection of maize aldolase 1 (ald1) gene (EU-RL GMFF GMOMETHODS database as of July 14, 2016). Estimated number of target copies per haploid genome (Jacchia et al. 2018): 2	ring trial validation	EU reference method	1
QT-TAX-ZM-005	taxon-specific	Zea mays	Quantitative PCR method for detection of maize high-mobility-group (hmg) gene (EU-RL GMFF GMOMETHODS database as of May 28, 2018).	ring trial validation	EU reference method	1
QT-TAX-ZM-006	taxon-specific	Zea mays	Quantitative real-time PCR (TaqMan) method for detection of maize starch synthase IIb gene (ISO 21570)	ring trial validation	ISO standard	1
QT-TAX-ZM-008	taxon-specific	Zea mays	Quantitative real-time PCR method for detection of maize high-mobility-group (hmg) gene (EU-RL GMFF GMOMETHODS database as of July 20, 2023). Estimated number of target copies per haploid genome (Jacchia et al. 2018): 1	ring trial validation	EU reference method	1
QT-TAX-ZM-011	taxon-specific	Zea mays	Quantitative real-time PCR (TaqMan) method for detection of the maize alcohol dehydrogenase 1 gene (EU-RL GMFF GMOMETHODS database as of March 06, 2014)	ring trial validation	EU reference method	1
QT-TAX-ZM-014	taxon-specific	Zea mays	Quantitative real-time PCR (TaqMan) method for detection of maize alcohol dehydrogenase 1 gene (EU-RL GMFF GMOMETHODS database as of March 06, 2014). Estimated number of target copies per haploid genome (Jacchia et al. 2018): 1	ring trial validation	EU reference method	1
QT-TAX-zSSIIb-3F/zSSIIb-4R	taxon-specific	Zea mays	Quantitative real-time PCR (TaqMan) method for detection of maize starch synthase IIb gene (Yang et al., 2005)	unknown	unknown	1

Catalogue number	Source	Value	Type	Certified	Available
ERM-BF445a	JRC - CRM (ERM)	0 % (m/m)	dried powder	yes	yes
ERM-BF445b	JRC - CRM (ERM)	98.7 % (m/m)	dried powder	yes	yes

Citation	Publication year	Publishing authority	Publication type	Content types
Assessment of genetically modified maize DP23211 for food and feed uses, under Regulation (EC) No 1829/2003 (application EFSA-GMO-NL-2019-163)	2024	EFSA GMO Panel	Scientific opinion EFSA GMO Panel	risk assessment document
EU-RL GMFF GMOMETHODS database; QT-EVE-ZM-035	2024	JRC	EU reference method protocol	detection
Application for authorisation of genetically modified plants DP23211 maize and derived food and feed in accordance with regulation (EC) No 1829/2003, EFSA-GMO-NL-2019-163, Part VII, Summary	2019	EFSA	application summary	application summary

European GMO Initiative for a Unified Database System

GMO detailed information

GMO:

Alias:

UID:

Tradename:

Companies/Institutions:

Species:

Traits:

EU authorisation:

General Information

Links regarding to this GMO:

Inserts / Modification

Transformation Vectors

Related DNA Sequences

Detection Methods

Reference Materials

Authorisation

Literature reference