CA2305864A1 - Transgenic reduction of sinapine in crucifera - Google Patents
Transgenic reduction of sinapine in crucifera Download PDFInfo
- Publication number
- CA2305864A1 CA2305864A1 CA 2305864 CA2305864A CA2305864A1 CA 2305864 A1 CA2305864 A1 CA 2305864A1 CA 2305864 CA2305864 CA 2305864 CA 2305864 A CA2305864 A CA 2305864A CA 2305864 A1 CA2305864 A1 CA 2305864A1
- Authority
- CA
- Canada
- Prior art keywords
- sequence
- seq
- plant
- cyp84
- promoter
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The sinapine content of seeds of Brassica napus, and other crucifera plants, and the resulting seed meal made therefrom, is reduced by transforming cells of the plants to incorporate an expressible exogenous CYP84 monooxygenase enzyme, particularly ferulate 5-hyroxylase (FSH) enzyme, or an antisense equivalent thereof. This allows for the production of a seed meal that is commercially more valuable. Three specific nucleic acid sequences encoding the FSH polypeptide are disclosed, designated BNFSH1, BNFSH2 and BNFSH3, and genetic constructs produced. The antisense suppression of sinapine is preferred, which can reduce the sinapine content of seed meal by up to 40%
compared to wild type or vector-only transformed plants.
compared to wild type or vector-only transformed plants.
Claims (48)
1. A transformed seed-producing plant, or a part thereof, of the Crucifera family containing an exogenous DNA sequence operably linked to a promoter for expression of a polypeptide that acts as a CYP84 monooxygenase enzyme, or a polypeptide that is an antisense equivalent of a CYP84 monooxygenase enzyme, or a polypeptide that is a catalytic fragment or derivative of said monooxygenase enzyme or antisense equivalent that does not alter the function of said CYP84 enzyme or antisense equivalent, and which plant or plant part has a reduced content of sinapine in seeds thereof compared to vector control plants or plant parts of the same species lacking said exogenous DNA sequence.
2. The plant or part thereof of claim 1, wherein the sequence expresses an antisense equivalent of a CYP84 monooxygenase or a catalytic fragment or derivative thereof.
3. The plant or part thereof of claim 2, wherein the sequence expresses an antisense equivalent of ferulate 5-hyroxylase or a catalytic fragment or derivative thereof.
4. The plant or part thereof of claim 1, wherein the sequence expresses a monooxygenase in sense orientation or a catalytic fragment or derivative thereof.
5. The plant or part thereof of claim 4, wherein the sequence expresses a ferulate 5-hydroxylase enzyme.
6. The plant or part thereof of claim 1, being of a species of Brassicaceae.
7. The plant or part thereof of claim 6, being of a species selected from the group consisting of Brassica napus, Brassica raps, Brassica oleracea, Brassica juncea, Brassica nigra and Sinapsis alba.
8. The plant or part thereof of claim 1, being of a species selected from the group consisting of Brassica napus, Brassica rapa and Brassica oleracea.
9. The plant or part thereof of claim 1, being of the species Brassica napus.
10. The plant or part thereof of claim 1, wherein the exogenous DNA sequence is selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2 and SEQ ID
NO:3, or is a sequence complementary to a sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, or is an analogue, homologue or derivative thereof, or a sequence having at least 40% identity with a sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, or at least 40% identity with a sequence complementary thereto.
NO:3, or is a sequence complementary to a sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, or is an analogue, homologue or derivative thereof, or a sequence having at least 40% identity with a sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, or at least 40% identity with a sequence complementary thereto.
11. A method of producing a transformed seed-producing plant, or part thereof, of the Crucifera family having a reduced content of sinapine in seeds thereof compared to vector control plants of the same species lacking said exogenous DNA
sequence, said method comprising transforming cells of said plant or plant part with an exogenous DNA sequence operably linked to a promoter for expression in said plant cell of a polypeptide that acts as a CYP84 monooxygenase enzyme, or a polypeptide that is an antisense equivalent of an CYP84 monooxygenase enzyme, or a polypeptide that is a catalytic fragment or derivative of said monooxygenase enzyme or antisense equivalent, and selecting transformed plants or plant parts having reduced content of sinapine in seeds thereof compared to vector control plants or plant parts of the same species lacking said exogenous DNA sequence.
sequence, said method comprising transforming cells of said plant or plant part with an exogenous DNA sequence operably linked to a promoter for expression in said plant cell of a polypeptide that acts as a CYP84 monooxygenase enzyme, or a polypeptide that is an antisense equivalent of an CYP84 monooxygenase enzyme, or a polypeptide that is a catalytic fragment or derivative of said monooxygenase enzyme or antisense equivalent, and selecting transformed plants or plant parts having reduced content of sinapine in seeds thereof compared to vector control plants or plant parts of the same species lacking said exogenous DNA sequence.
12. The method of claim 11, wherein the sequence expresses an antisense equivalent of a CYP84 monooxygenase or a catalytic fragment or derivative thereof.
13. The method of claim 12, wherein the sequence expresses an antisense equivalent of ferulate 5-hyroxylase or a catalytic fragment or derivative thereof.
14. The method of claim 11, wherein the sequence expresses a CYP84 monooxygenase in sense orientation or a catalytic fragment or derivative thereof.
15. The method of claim 14, wherein the sequence expresses a ferulate 5-hydroxylase enzyme.
16. The method of claim 11, wherein the plant or part thereof is of a species of Brassicaceae.
17. The method of claim 16, wherein the plant or part thereof is of a species selected from the group consisting of Brassica napus, Brassica rapa, Brassica oderacea, Brassica juncea, Brassica nigra and Sinapsis alba.
18. The method of claim 11, wherein the plant or part thereof is of a species selected from the group consisting of Brassica napus, Brassica rapa and Brassica oleracea.
19. The method of claim 11, wherein the plant or part thereof is of the species Brassica napus.
20. The method of claim 11, wherein the endogenous DNA sequence is selected from SEQ ID NO: 1, SEQ ID NO:2 and SEQ ID NO:3, or a is a sequence complementary to a sequence selected from the group consisting of SEQ ID
NO:1, SEQ ID NO:2 and SEQ ID NO:3, or is an analogue, homologue or derivative thereof.
NO:1, SEQ ID NO:2 and SEQ ID NO:3, or is an analogue, homologue or derivative thereof.
21. A seed meal prepared by growing the plant or part thereof of claim 1, harvesting seeds from the plant, and processing said seeds to form seed meal.
22. An isolated nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3 or SEQ ID NO:5, or an active fragment, derivative, homolog or analog thereof, or a sequence that is complementary to SEQ ID NO:1, SEQ ID NO:3 or SEQ ID NO:5, or an active fragment thereof, derivative, homolog or analog thereof., or a sequence having at least 90% identity to said sequence or complementary sequence.
23. The isolated nucleic acid molecule of claim 22, comprising the sequence of nucleotides of SEQ ID NO:1.
24. The isolated nucleic acid molecule of claim 22, comprising the sequence of nucleotides of SEQ ID NO:3.
25. The isolated nucleic acid molecule of claim 22, comprising the sequence of nucleotides SEQ ID NO:5.
26. The isolated nucleic acid molecule of claim 22, comprising a sequence that is complementary to SEQ ID NO: 1.
27. The isolated nucleic acid molecule of claim 22, comprising a sequence of nucleotides that is complementary to SEQ ID NO:3.
28. The isolated nucleic acid molecule of claim 22, comprising a sequence of nucleotides that is complementary to SEQ ID NO:5.
29. An expressed and isolated polypeptide molecule comprising a sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO:4 and SEQ ID NO:6, or an active fragment, homolog, analog or derivative thereof.
30. A vector for transforming plants of the crucifera family, said vector comprising a promoter, a start codon, a sequence operably linked to said promoter, selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3, and SEQ ID NO:5, or a sequence complementary thereto, or an active fragment, homolog, analog or derivative of said sequence or complementary sequence, and a stop codon.
31. The vector of claim 30, wherein the promoter is the CaMV 35S promoter.
32. The vector of claim 30, wherein the promoter is the napin promoter.
33. Plasmid identified by Accession no. IDAC 020500-5.
34. Plasmid identified by Accession no. IDAC 020500-6.
35. Plasmid identified by Accession no. IDAC 020500-7.
36. A transgenic Brassica napus plant, or part thereof, transformed with the vector of claim 30.
37. A chimeric gene causing reduced sinapine content in plant cells of the Crucifera family transformed with the chimeric gene, the chimeric gene comprising:
a regulatory nucleotide sequence; and a nucleic acid fragment encoding an active plant CYP84 enzyme; and wherein the nucleic acid fragment is operably linked in either the sense or antisense orientation to the regulatory sequence.
a regulatory nucleotide sequence; and a nucleic acid fragment encoding an active plant CYP84 enzyme; and wherein the nucleic acid fragment is operably linked in either the sense or antisense orientation to the regulatory sequence.
38. The chimeric gene of claim 37 wherein the CYP84 enzyme is a F5H enzyme.
39. The chimeric gene of claim 37 wherein the CYP84 enzyme has an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4 or SEQ ID NO:6.
40. The chimeric gene of claim 37 wherein the nucleic acid fragment is operably linked in the sense orientation to said regulatory sequence.
41. The chimeric gene of claim 37 wherein the nucleic acid fragment is operably linked in the antisense orientation to said regulatory sequence.
42. The chimeric gene of claim 37 wherein the regulatory sequence comprises a promoter selected from the group consisting of a napin promoter, a phaseolin promoter, an oleosin promoter and a cruciferin promoter.
43. The chimeric gene of claim 37 wherein the regulatory sequence comprises an endogenous plant promoter effective for controlling expression of a plant gene.
44. A seed meal comprising seeds, or parts thereof, of a transformed seed-producing plant of the Crucifera family containing an exogenous DNA sequence operably linked to a promoter for expression of a polypeptide that acts as a CYP84 monooxygenase enzyme, or a polypeptide that is an antisense equivalent of an CYP84 monooxygenase enzyme, or a polypeptide that is a catalytic fragment or derivative of said monooxygenase enzyme or antisense equivalent that does not alter the function of said CYP84 enzyme or antisense equivalent, and which plant or plant part has a reduced content of sinapine in seeds thereof compared to vector control plants or plant parts of the same species lacking said exogenous DNA
sequence.
sequence.
45. The seed meal of claim 44 wherein said transformed plant is canola.
46. A method of producing a transformed seed-producing plant, or part thereof, of the Crucifera family having a reduced content of sinapine in seeds thereof compared to vector control plants of the same species lacking said exogenous DNA
sequence, said method comprising transforming cells of said plant or plant part with an exogenous DNA sequence operably linked to a promoter for expression in said plant cell of a polypeptide, said DNA sequence being a sequence flanking an endogenous CYP84 gene of the plant, or a sequence that is complementary thereto.
sequence, said method comprising transforming cells of said plant or plant part with an exogenous DNA sequence operably linked to a promoter for expression in said plant cell of a polypeptide, said DNA sequence being a sequence flanking an endogenous CYP84 gene of the plant, or a sequence that is complementary thereto.
47. The method of claim 46, wherein said CYP84 gene is an F5H gene.
48. An antibody molecule that binds specifically to a polypeptide of the group consisting fo SEQ ID NO:2, SEQ ID NO:4 and SEQ ID NO:6, or a polypeptide having an amino acid sequence with at least 40% identity thereto.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA 2305864 CA2305864C (en) | 1999-05-06 | 2000-05-05 | Transgenic reduction of sinapine in crucifera |
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US13280099P | 1999-05-06 | 1999-05-06 | |
| CA002270417A CA2270417A1 (en) | 1999-05-06 | 1999-05-06 | Transgenic reduction of the sinapine content in brassica napus seeds |
| CA2,270,417 | 1999-05-06 | ||
| US60/132,800 | 1999-05-06 | ||
| CA 2305864 CA2305864C (en) | 1999-05-06 | 2000-05-05 | Transgenic reduction of sinapine in crucifera |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2305864A1 true CA2305864A1 (en) | 2000-11-06 |
| CA2305864C CA2305864C (en) | 2010-07-20 |
Family
ID=27170960
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA 2305864 Expired - Fee Related CA2305864C (en) | 1999-05-06 | 2000-05-05 | Transgenic reduction of sinapine in crucifera |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA2305864C (en) |
-
2000
- 2000-05-05 CA CA 2305864 patent/CA2305864C/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CA2305864C (en) | 2010-07-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP0480024B1 (en) | Dna encoding plant thioesterases | |
| US7166771B2 (en) | Coordinated decrease and increase of gene expression of more than one gene using transgenic constructs | |
| AU708654B2 (en) | DNA molecules which code for a plastid 2-oxoglutarate/malate translocator | |
| CA2257198C (en) | Suppression of specific classes of soybean seed protein genes | |
| EP1967589B1 (en) | An oleosin 5' regulatory region for the modification of plant seed lipid composition | |
| US7157621B2 (en) | Alteration of oil traits in plants | |
| Zhang et al. | Expression of antisense or sense RNA of an ankyrin repeat-containing gene blocks chloroplast differentiation in arabidopsis. | |
| AU753176B2 (en) | Specific gene activation by chimeric Gal4 transcription factors in stable transgenic plants | |
| JPH06506584A (en) | Plant promoters involved in controlling lipid biosynthesis in seeds | |
| CA2268860A1 (en) | Leaf-specific gene expression in transgenetic plants | |
| US6825335B1 (en) | Synthetic fatty acid desaturase gene for expression in plants | |
| CA2919816C (en) | Gene capable of increasing seed protein content and method of use thereof | |
| US20160032307A1 (en) | Modifying the fatty acid profile of camelina sativa oil | |
| JP6224030B2 (en) | Methods for modifying fructan biosynthesis, increasing plant biomass, and enhancing biochemical pathway productivity in plants | |
| CA2381316A1 (en) | Homogentisate-dioxygenase | |
| AU778179B2 (en) | Gibberellin 3 beta-hydroxylase genes of rice and uses thereof | |
| Alonso et al. | A non-photosynthetic ferredoxin gene is induced by ethylene in Citrus organs | |
| US6025544A (en) | Processes for modifying plant flowering behavior | |
| WO2005080579A2 (en) | Taxadiene biosynthesis | |
| CA2305864A1 (en) | Transgenic reduction of sinapine in crucifera | |
| KR102000471B1 (en) | Constitutive promoter derived from chrysanthemum and use thereof | |
| US6501004B1 (en) | Transgenic reduction of sinapine in crucifera | |
| AU765413B2 (en) | Light-repressible promoters | |
| US6469149B1 (en) | Production of ascorbic acid in plants | |
| EP1298214B1 (en) | Plant thioesterases |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| MKLA | Lapsed |