GMO molecular characterization details

GMO detailed information

GMO:
PY203
Alias:
UID:
AGV-PY2Ø3-4
Tradename:

GraINzyme Phytase
Companies/Institutions:
Agrivida
Species:
Zea mays
(maize, corn)
Traits:
EU authorisation:

no Reference materials could be found
No results found.
no Literature reference could be found

General Information

Description:

Maize Event PY203 was generated by Agrobacterium-mediated transformation into immature maize embryo tissue with the transformation vector pAG4758.

PY203 maize plants express the phytase gene phy02 , a modified appA phytase gene derived from Escherichia coli strain K12.

Phytases catalyze the step-wise removal of phosphate from the plant produced phosphate storage molecule phytin or phytic acid. They are commonly added to the diets of monogastric animals to increase the nutritional availability of phosphate. PY203 was developed to be added as ground material to animal feed to improve phosphorus utilization.

The transformation vector pAG4758 contains:

• three copies of the phy02 gene (CS-phy02-ECOLX), each with a different monocot derived promoter, the Oryza sativa glutelin-1 gene promoter (P-gluA1-ORYSA), the Zea mays Zc2 gene promoter (P-zc2-MAIZE) and the Zea mays globulin Glb1 gene promoter (P-glb1-MAIZE), and the nos terminator (T-nos-RHIRD),

• the pmi gene (CS-pmi-ECOLX) encoding the phosphomannose isomerase (PMI) enzyme as a plant selectable marker with the Zea mays ubiquitin 1 gene promoter (P-ubi1-MAIZE) and the nos terminator (T-nos-RHIRD).

Data from Southern analyses and DNA sequencing demonstrated that two independent copies of the T-DNA from the transformation construct pAG4758 are inserted into the genome of Event PY203. Both insertions were demonstrated to be stable over several generations.

Southern blots and PCR confirmed that the maize Event PY203 does not contain any vector backbone sequences of the transformation plasmid pAG4758.

Links regarding to this GMO:

FDA https://www.fda.gov/files/combined%20redacted%20notice%20AGRN%2021.pdf,

USDA APHIS https://www.aphis.usda.gov/brs/aphisdocs/19_17601p.pdf,

Updated APHIS petition, Regulations.gov https://downloads.regulations.gov/APHIS-2019-0084-0002/content.pdf,

BCH https://bch.cbd.int/en/database/record?documentID=115832

Transformation / Modification technique:

Agrobacterium-induced

Inserts / Modification

Name:	insert1_PY203
Comment:	Insert 1 on chromosome 8 (locus 3293) contains a complete copy of the T-DNA with three copies of the phy02 genes and a single copy of the phosphomannose isomerase (pmi) gene. It lacks 27 bp from the RB end of the T-DNA, including the entire 25 bp RB element.
Map:

Genetic Elements:

Reference: USDA-APHIS

Elements:

Order	Name	Donor	Position	Definition	Trait	Comment
1	V-Flanking Ti-nos-RHIRD	Agrobacterium tumefaciens	-	Flanking sequence from Agrobacterium tumefaciens Ti plasmids of the nopaline type; sequence located in the promoter of nopaline synthase.		TI-plasmid right border repeat (RB)
2	P-gluA1-ORYZA	Oryza sativa	-	Promoter derived from Oryza sativa Glutelin-1 gene (similar to GenBank accession number EU264103.1). Qu et al. (2008). J Exp Botany, 59, 2417-2424		length: 2071 bp
3	TP-zc2-MAIZE	Zea mays	-	Signal peptide sequence derived from the 27 kDA-zein seed storage protein of Zea mays (similar to Genbank accession number AB086264.1). Torrent et al. (2009). MBC Biology, 7, 5		length: 57 bp
4	CS-phy02-ECOLX	Escherichia coli	-	phy02 is a modified version of the E.coli appA phytase gene (Genbank accession number EFE63517.1).	Phytase	length: 1257 bp
5	R-SEKDEL-SYNTH	Synthetic	-
6	T-nos-RHIRD	Agrobacterium tumefaciens	-	3' untranslated region (3' UTR) of the nopaline synthase gene from Agrobacterium tumefaciens T-DNA. Bevan et al. (1983) Nucleic Acids Res. 11: 369-385; Fraley et al. (1983) Proc Nat L Acad Sci. 80:4803-4807
7	P-zc2-MAIZE	Zea mays	-	Promoter sequence derived from Z. mays Zc2 gene (GenBank accession number X53514.1). Reina et al. (1990). Nucleic Acids Res, 18, 6425		length: 1363 bp
8	TP-zc2-MAIZE	Zea mays	-	Signal peptide sequence derived from the 27 kDA-zein seed storage protein of Zea mays (similar to Genbank accession number AB086264.1). Torrent et al. (2009). MBC Biology, 7, 5		length: 57 bp
9	CS-phy02-ECOLX	Escherichia coli	-	phy02 is a modified version of the E.coli appA phytase gene (Genbank accession number EFE63517.1).	Phytase	length: 1257 bp
10	R-SEKDEL-SYNTH	Synthetic	-
11	T-nos-RHIRD	Agrobacterium tumefaciens	-	3' untranslated region (3' UTR) of the nopaline synthase gene from Agrobacterium tumefaciens T-DNA. Bevan et al. (1983) Nucleic Acids Res. 11: 369-385; Fraley et al. (1983) Proc Nat L Acad Sci. 80:4803-4807
12	P-glb1-MAIZE	Zea mays	-	Promoter region of the globulin 1 gene.	Altered amino acid composition
13	TP-zc2-MAIZE	Zea mays	-	Signal peptide sequence derived from the 27 kDA-zein seed storage protein of Zea mays (similar to Genbank accession number AB086264.1). Torrent et al. (2009). MBC Biology, 7, 5		length: 57 bp
14	CS-phy02-ECOLX	Escherichia coli	-	phy02 is a modified version of the E.coli appA phytase gene (Genbank accession number EFE63517.1).	Phytase	length: 1257 bp
15	R-SEKDEL-SYNTH	Synthetic	-
16	T-nos-RHIRD	Agrobacterium tumefaciens	-	3' untranslated region (3' UTR) of the nopaline synthase gene from Agrobacterium tumefaciens T-DNA. Bevan et al. (1983) Nucleic Acids Res. 11: 369-385; Fraley et al. (1983) Proc Nat L Acad Sci. 80:4803-4807
17	P-ubi1-MAIZE	Zea mays	-	Promoter region of the polyubiquitin gene. Christensen et al. (1992). Plant Mol Biol, 18, 675-689
18	CS-pmi-ECOLX	Escherichia coli	-	Phosphomannose isomerase. Reed et al (2001). In Vitro Cell Dev Biol Plant, 37, 127-132	Mannose metabolism
19	T-nos-RHIRD	Agrobacterium tumefaciens	-	3' untranslated region (3' UTR) of the nopaline synthase gene from Agrobacterium tumefaciens T-DNA. Bevan et al. (1983) Nucleic Acids Res. 11: 369-385; Fraley et al. (1983) Proc Nat L Acad Sci. 80:4803-4807
20	V-LB-RHIRD	Agrobacterium tumefaciens	-

Related DNA Sequences:

Acc.No.	Reference	Sequence	Size (bp)	Annotation
	USDA-APHIS	No
US_11371052_B2_42	patent documents	Yes	18621	Sequence of the T-DNA insertion on locus 3293 (Chromosome 8) including T-DNA and maize host genome flanking DNA

Name:	insert2_PY203
Comment:	Insert 2 on chromosome 2 (locus 3507) is a truncated version of the T-DNA that includes two of the three phy02 genes from the T-DNA fragment. This insertion lacks the third, downstream copy of the phy02 gene and much of the ZmGlb1 promotor from which the gene is expressed as well as the entire pmi gene.
Map:

Genetic Elements:

Reference: USDA-APHIS

Elements:

Order	Name	Donor	Position	Definition	Trait	Comment
1	V-Flanking Ti-nos-RHIRD	Agrobacterium tumefaciens	-	Flanking sequence from Agrobacterium tumefaciens Ti plasmids of the nopaline type; sequence located in the promoter of nopaline synthase.		TI-plasmid right border repeat (RB)
2	P-gluA1-ORYZA	Oryza sativa	-	Promoter derived from Oryza sativa Glutelin-1 gene (similar to GenBank accession number EU264103.1). Qu et al. (2008). J Exp Botany, 59, 2417-2424		length: 2071 bp
3	TP-zc2-MAIZE	Zea mays	-	Signal peptide sequence derived from the 27 kDA-zein seed storage protein of Zea mays (similar to Genbank accession number AB086264.1). Torrent et al. (2009). MBC Biology, 7, 5		length: 57 bp
4	CS-phy02-ECOLX	Escherichia coli	-	phy02 is a modified version of the E.coli appA phytase gene (Genbank accession number EFE63517.1).	Phytase	length: 1257 bp
5	R-SEKDEL-SYNTH	Synthetic	-
6	T-nos-RHIRD	Agrobacterium tumefaciens	-	3' untranslated region (3' UTR) of the nopaline synthase gene from Agrobacterium tumefaciens T-DNA. Bevan et al. (1983) Nucleic Acids Res. 11: 369-385; Fraley et al. (1983) Proc Nat L Acad Sci. 80:4803-4807
7	P-zc2-MAIZE	Zea mays	-	Promoter sequence derived from Z. mays Zc2 gene (GenBank accession number X53514.1). Reina et al. (1990). Nucleic Acids Res, 18, 6425		length: 1363 bp
8	TP-zc2-MAIZE	Zea mays	-	Signal peptide sequence derived from the 27 kDA-zein seed storage protein of Zea mays (similar to Genbank accession number AB086264.1). Torrent et al. (2009). MBC Biology, 7, 5		length: 57 bp
9	CS-phy02-ECOLX	Escherichia coli	-	phy02 is a modified version of the E.coli appA phytase gene (Genbank accession number EFE63517.1).	Phytase	length: 1257 bp
10	R-SEKDEL-SYNTH	Synthetic	-
11	T-nos-RHIRD	Agrobacterium tumefaciens	-	3' untranslated region (3' UTR) of the nopaline synthase gene from Agrobacterium tumefaciens T-DNA. Bevan et al. (1983) Nucleic Acids Res. 11: 369-385; Fraley et al. (1983) Proc Nat L Acad Sci. 80:4803-4807
12	P-glb1-MAIZE	Zea mays	-	Promoter region of the globulin 1 gene.	Altered amino acid composition	truncated

Related DNA Sequences:

Acc.No.	Reference	Sequence	Size (bp)	Annotation
	USDA-APHIS	No
US_11371052_B2_43	patent documents	Yes	13109	Sequence of the T-DNA insertion on locus 3507 (Chromosome 2) including truncated T-DNA and maize host genome flanking DNA

Detection Methods

21 results

Name	Type	Target	Description	Validation	Standardisation	Verification
QL-ELE-AgroBS1 F/AgroBS1 R	element-specific	V-Flanking Ti-nos-RHIRD	Qualitative real-time PCR (TaqMan) method for detection of the Agrobacterium tumefaciens Ti plasmids of the nopaline type aka Agroborder I. Part of the qualitative triplex real-time PCR (TaqMan) method for detection of Ti plasmid (AgroBorder) sequences and a sequence of the Cassava vein mosaic virus promoter pat construct (P-CsVMV-pat) for screening for components of genetically modified organisms in food (Official Collection of Methods according to § 64 LFGB, L 00.00-176)	ring trial validation	national standard	2
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-00-006	element-specific	T-nos-RHIRD	Qualitative conventional PCR method for detection of nopaline synthase terminator (Official Collection of Methods according to § 64 LFGB, L 00.00-31)	ring trial validation	national standard	1
QL-ELE-00-007	element-specific	T-nos-RHIRD	Qualitative conventional PCR method for detection of nopaline synthase terminator (EU-Project SMT4-CT96-2072:1998)	ring trial validation	national standard	1
QL-ELE-00-009	element-specific	T-nos-RHIRD	Qualitative conventional PCR method for detection of nopaline synthase terminator (Lipp et al.,2001)	ring trial validation	ISO standard	1
QL-ELE-00-011	element-specific	T-nos-RHIRD	Qualitative real-time PCR (TaqMan) method for detection of nopaline synthase terminator (ISO 21569 AMD1).	ring trial validation	ISO standard	1
QL-ELE-00-013	element-specific	T-nos-RHIRD	Qualitative real-time PCR (TaqMan) method for detection of nopaline synthase terminator (partim T-nos). Part of the qualitative duplex real-time PCR (TaqMan) method for detection of Cauliflower mosaic virus 35S promoter and nopaline synthase terminator (partim T-nos) (ISO 21569 AMD1)	ring trial validation	ISO standard	1
QL-ELE-00-018	element-specific	T-nos-RHIRD	Qualitative real-time PCR (SYBRGreen) method for detection of T-nos-RHIRD (Barbau-Piednoir et al., 2014)	ring trial validation	unknown	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-ELE-NOS ter 2-5' /NOS ter 2-3'	element-specific	T-nos-RHIRD	Quantitative real-time PCR (TaqMan) method for detection of T-nos (Kodama et al., 2009)	ring trial validation	none	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

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

Detection Methods

Reference Materials

Authorisation

Literature reference