Brassica juncea line NUBJ1207

Abstract

The present embodiments provide a novel DPA-producing Brassica juncea (DPA juncea) line designated NUBJ1207. The embodiments also provide seeds, plants, and plant parts (including DNA) of the DPA juncea line NUBJ1207, as well as methods for generating Brassica plants produced by crossing the DPA juncea line NUBJ1207 with itself or another Brassica line. These embodiments also provide methods for generating DPA-producing Brassica plants that contain one or more transgenes within their genetic material, and transgenic plants and transgenic plant parts produced by these methods. These embodiments further provide methods for generating DPA-producing lines or DPA breeding lines and plant parts derived from the DPA juncea line NUBJ1207, methods for generating canola lines or plant parts derived from line NUBJ1207, and DPA-producing Brassica plants, subspecies, and parts thereof derived from the use of these methods. These embodiments also provide hybrid seeds, plants, and plant parts produced by crossing DPA juncea line NUBJ1207 with another Brassica line or a canola line.

Description

Technical Field

[0001] Related Applications This application claims the benefit of priority of U.S. Provisional Patent Application No. 62 / 891,908, filed Aug. 26, 2019, which is hereby incorporated by reference in its entirety for all purposes.

[0002] Sequence Listing This application contains a sequence listing (created on Jun. 20, 2019, first submitted as a 2,914-byte file named 87376_0015PRO_ST25.txt via EFS-Web in ASCII format on Aug. 26, 2019), which is filed herewith simultaneously with the specification and is hereby incorporated by reference in its entirety.

[0003] This embodiment relates to the novel and specific DPA-producing Brassica juncea line designated NUBJ1207.

Background Art

[0004] Long-chain omega-3 polyunsaturated fatty acids (ω3 LC PUFAs) are widely recognized for their important roles in the human diet. ω3 LC PUFA Δ7,10,13,14,17-docosapentaenoic acid (DPA) contributes to various aspects of human healthy life, from heart and vascular health to brain development and lifelong brain function. A land-based source of DPA is needed.

Summary of the Invention

[0005] This embodiment provides a plant, derivative or progeny plant, a part of the plant, or seeds of the DPA Brassica juncea line NUBJ1207 (the "DPA juncea"), and a representative sample of the seeds has been deposited under ATCC Accession No. PTA-125954 (see the appendix). One aspect of the embodiment provides a method for generating DPA juncea seeds, the method including crossing two Brassica plants and harvesting the resulting seeds, where at least one of the two Brassica plants is the DPA juncea line NUBJ1207 or its progeny.

Embodiments for Carrying Out the Invention

[0006] It should be noted that the present invention is not limited to the specific methods, protocols, and reagents described herein and may vary accordingly. The terms used herein are for the purpose of describing only specific embodiments and are not intended to limit the scope of the present invention, which is defined only by the claims.

[0007] All patents or other publications are hereby incorporated by reference into this specification for the purpose of, for example, explaining and disclosing the methods described in such publications that may be used in connection with the present invention, but are not to be construed as providing definitions of terms that are inconsistent with the definitions presented herein. Such publications are provided only for the purpose of disclosing prior to the filing date of this application. In this regard, nothing should be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or for any other reason. All statements as to the date or content of such documents are based on the information available to the applicant and do not constitute an admission as to the accuracy of the date or content of such documents.

[0008] As used in this specification and the claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Throughout this specification, unless otherwise specified, the terms "comprise", "comprises", and "comprising" are used in a non-exclusive and inclusive sense, such that the recited integer or group of integers may include one or more other unrecited integers or groups of integers. The term "or" is inclusive, unless modified, for example, by "either". Thus, unless the context otherwise dictates, the word "or" means any one member of a particular list and also includes any combination of members of that list.

[0009] All values are approximate because there is some variation due to environmental conditions in the fatty acid compositions. Values are generally expressed as area percentages, which approximate the weight percentages of the total fatty acids, or of the total seeds. Thus, unless otherwise indicated in an example or otherwise specified, all numbers expressing amounts or reaction conditions used in this specification are to be understood as being modified in all instances by the term "about".

[0010] Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art. The terms used in this specification are for the purpose of describing particular embodiments only and are not intended to limit the scope of the invention, which is defined only by the claims. To better understand the present disclosure, certain terms are first defined. Further definitions are set forth throughout the detailed description.

[0011] This embodiment provides a Brassica line that produces LC ω3 PUFA DPA in its seed oil, and in particular, provides a line that embodies the characteristics of the B. juncea line NUBJ1207. LC ω3 PUFAs are known in the art by appropriate discussion thereof and for other fatty acids; see, for example, U.S. Patent No. 10,570,405.

[0012] Brassica juncea belongs to the Brassicaceae (Cruciferae) plant family, commonly known as the mustard family. Generally, B. juncea has light green leaves with a few hairs on the first leaf and the leaf blade that remains on the petiole. The plant reaches a height of 1-2 meters, has deep lobes on the lower leaves, and the upper leaves are narrow and open overall and do not close. The flowers are pale yellow and open gradually upward from the bottom of the slender raceme. The silique (seed pod) is slightly adherent and is 2.5-5 cm long excluding the beak, which is 0.5-1 cm long, and the seeds are round and may be yellow or brown. B. juncea is an amphidiploid (AABB genome, 2n = 36) of B. rapa (AA genome, 2n = 20) by B. nigra (BB genome, 2n = 16).

[0013] Brassica juncea ssp. juncea is cultivated mainly for its seeds. More specifically, the oilseed B. juncea is grown as a spice crop (mustard seed) in North America, but is also used as a source of cooking oil in Asia and other regions. Almost all of the oilseed B. juncea grown (mostly in the Indian subcontinent) has high levels of both erucic acid and glucosinolates, but there are low glucosinolate and low erucic acid subspecies that provide canola-quality edible oil. More specifically, "canola oil" is defined as having less than 2% erucic acid, and canola meal is defined as non-oil meal, air-dried to 1 gram, containing less than 30 μmol of 3-butenyl glucosinolate, 4-pentenyl glucosinolate, 2-hydroxy-3-butenyl glucosinolate, 2-hydroxy-4-pentenyl glucosinolate, or mixtures thereof. See, for example, CODEX ALIMENTARIUS: FATS, OILS & RELATED PRODUCTS, Vol. 8 (2nd Ed., Food & Agriculture Org. United Nations, Rome, Italy, 2001). In particular, the erucic acid content of the seed oil of the B. juncea line NUBJ1207 is less than 1%.

[0014] Compared to canola species B. napus and B. rapa, which are grown over a wider range, B. juncea is more resistant to heat and drought stress. Typically, B. juncea is more resistant to crushing than some B. napus, and can be either banded or linear in combination. Further information on the ecology, hybridization, and diseases of B. juncea is readily available. See, for example, Canadian Food Inspection Agency, Biology Document BIO2007-01, Biology of Brassica juncea (Canola / Mustard) (2012). Neither wild-type B. juncea, B. napus, nor B. rapa produces DPA in seed oil.

[0015] A "line" is a group of plants that shows little overall variation among individuals sharing that designation. A "line" also refers to a homogeneous population of plants having substantially the same genetic material, which shows little or no genetic variation among individuals, particularly with respect to at least one trait and in the transgenes present in line NUBJ1207 and the resulting DPA product. The terms "subspecies" or "cultivar" may be used interchangeably with "line", although generally the former two terms refer to lines suitable for commercial production. For example, "derived genetically", as used in the phrase "derived genetically from the parental line", means that the characteristic in question is indicated in whole or in part by some aspect of the genetic constitution of the plant in question.

[0016] "Progeny" means all descendants, including the offspring and derivatives of a plant, and includes the first, second, third, and subsequent generations, and may be produced by self-pollination of the plant or by crossing with plants having the same or different genotypes, and may be modified by various suitable genetic engineering techniques. The cultivated variety is usually related to a plant intentionally altered and selected by humans. "T0" refers to the first generation of transformed plant material, "T1" refers to the seeds produced on the T0 plant, and the T1 seeds give rise to plants that produce T2 seeds, etc. for subsequent Tx progeny.

[0017] "A part of a plant" includes plant cells, plant organs, plant protoplasts, plant cell tissue cultures from which plants can be redifferentiated, plant callus, plant aggregates, and intact plant cells in plants or parts thereof such as embryos, pollen, ovules, seeds, sheaths, leaves, flowers, branches, fruits, stalks, roots, root tips, anthers, cotyledons, hypocotyls, radicles, single cells, propagules, cell cultures, tissue cultures, etc. Cotyledons are a type of seed leaf and small leaves are contained on the plant embryo. Cotyledons contain the food storage tissue of the seed. An embryo is a small plant contained within a mature seed. "Plant cell" also encompasses non-redifferentiable plant cells. The progeny, derivatives, variants, and mutants of redifferentiated plants are also within the scope of this embodiment if some of them contain one, two, or three of the nucleic acid molecules as exemplified by SEQ ID NOs: 1 and 2, SEQ ID NOs: 3 and 4, and SEQ ID NOs: 5 and 6, and the molecules have the characteristics of DPA JUNCEA NUBJ1207. This embodiment also relates to the use of the elite event NUBJ1207 transgene in plant cell cultures and tissue cultures. This embodiment includes plants and parts of plants from the elite event NUBJ1207 line, as well as other plants produced by the described methods.

[0018] At least one embodiment described herein provides a novel B. juncea line, designated NUBJ1207, which produces DPA in its seed oil. Other aspects of this embodiment include the seeds, plants, and parts of plants of DPA juncea NUBJ1207, and methods for producing canola plants produced by crossing DPA juncea NUBJ1207 with itself or with other canola genotypes, and the generation of mutants by mutagenesis or transformation of DPA juncea NUBJ1207. Thus, this embodiment includes any method of using line NUBJ1207, examples of which include selfing, backcrossing, hybrid production, and crossing into a population. All plants produced using line NUBJ1207 as a parent are within the scope of this embodiment. In at least one embodiment, the DPA juncea line NUBJ1207 is used in crosses with other different canola plants to produce seeds and plants of first-generation (F 1 ) hybrids having excellent characteristics. Thus, examples of members of the genus Brassica useful in practicing this embodiment include, in addition to B. juncea, B. napus, B. napobrassica, B. oleracea, B. carinata, B. rapa, and B. campestris, and any other plant belonging to the genus Brassica that enables breeding between Brassica species. Typically, an "oilseed plant" refers to any one of the species of B. napus, B. rapa (or campestris), or B. juncea.

[0019] The phenotypic characterization of the DPA juncea line NUBJ1207 includes, in addition to the production of DPA in the seed oil, the following.

[0020]

Table 1

[0021] The DPA juncea described in this specification may be of A line, B line, or R line. The A line is a male sterile line, B is similar to "A" in all characteristics but is a male fertile line, and R, when crossed with the B line, restores fertility in the F1 hybrid. The B line is sometimes called the maintenance line because it is used to maintain fertility. The restoring line is a pollinating subspecies that is used to pollinate a male sterile line to produce F1 progeny that are male fertile and can self-propagate to produce seeds. Inbred lines may enable the restoration of fertility to the progeny of the male sterile line with which they are crossed. Thus, the hybrid line may be grown from F1 seeds produced by crossing two inbred (pure line) lines, one of which may be male sterile.

[0022] For pure line selection, the progeny (self-pollination) by single plant selection are maintained separately and are not bulked together. A set of genetically homogeneous lines is the expected result. Thus, a pure line may be the progeny of a homozygous plant by single self-fertilization.

[0023] Accordingly, the present embodiment provides a method for introducing desirable traits into the DPA juncea line NUBJ1207, where the method comprises crossing a NUBJ1207 plant with a plant of another Brassica genotype comprising the desirable trait for generating progeny plants, where the desirable trait is selected from the group consisting of male sterility, herbicide tolerance, pest tolerance, modified fatty acid metabolism, modified carbohydrate metabolism, modified seed yield, modified oil percentage, modified protein percentage, modified lodging resistance, and resistance to bacterial, fungal, or viral diseases; selecting one or more progeny plants having the desirable trait for generating selected progeny plants; crossing the selected progeny plants with a NUBJ1207 plant for generating progeny plants by backcrossing; selecting progeny plants by backcrossing having substantially all of the desirable traits of the NUBJ1207 line, as well as physiological and morphological characteristics; and repeating these steps three or more times for generating selected fourth and subsequent generation progeny plants by backcrossing, comprising substantially all of the desirable traits of the NUBJ1207 line, as well as physiological and morphological characteristics. Plants produced by this method are included in this aspect of the embodiment, where the plants have substantially all of the desirable traits (i.e., DPA production) of the NUBJ1207 line, as well as physiological and morphological characteristics.

[0024] "Breeding" includes all methods of growing or propagating plants and includes all suitable conventional breeding techniques and artificial breeding techniques, in addition to crossing both interspecies and intraspecies and interline and intra-line. Desirable traits (e.g., the DPA trait of NUBJ1207) may be transferred to other canola lines or B. juncea lines, cultivars, or accessions, or may be transferred through conventional breeding methods, or may be transferred to other Brassica species such as B. napus and B. rapa through interspecies crossing. Both conventional breeding methods and interspecies crossing methods, as well as other methods of transferring genetic material between plants, are well known in the art.

[0025] B. juncea line NUBJ1207 was developed by Agrobacterium tumefaciens-mediated transformation with the gene construct pJP3416_GA7-modB (“modB”) and contains seven genes that can promote the accumulation of omega-3 fatty acids in seeds. Briefly, the modB construct contains genes encoding a Δ6 desaturase cloned from the microalga Micromonas pusilla, a Δ6-elongase and a Δ5 elongase cloned from the microalga Pyramimonas cordata, a Δ5 desaturase cloned from the microalga Pavlova salina, a Δ15 / ω3-desaturase cloned from the yeast Pichia pastoris, a Δ12 desaturase cloned from the yeast Lachancea kluyveri, and a Δ4 desaturase cloned from Pavlova salina, each having an appropriate transcriptional promoter and terminator. See, for example, U.S. Patent Nos. 10,570,405 (FIG. 1), 10,563,218, 10,125,084, 9,718,759, 9,932,541. However, upon insertion into B. juncea (low erucic acid, low glucosinolate line) from which NUBJ1207 is derived, the Δ4-desaturase gene of modB was truncated and rendered non-functional.

[0026] Genetic analysis revealed that NUBJ1207 contains the loci of three recombinant transgenes, each of which has a separate transgene insert containing an expression cassette, and each insert is located in a different chromosome. In particular, the transgene insert in chromosome A06 of NUBJ1207 contains a truncated MAR sequence (640 bp Rb7 matrix attachment region of Nicotiana tabacum), a promoter of Arabidopsis thaliana FA elongase 1 (Pro_Arath-FAE1), the P. cordata Δ6-elongase gene, a terminator of Glycine max lectin (Ter_Glyma-Lectin), a promoter of cauliflower mosaic virus 35S RNA, a coding sequence of phosphinothricin acetyltransferase (PAT) enzyme, a terminator from A. tumefaciens nopaline synthase (TER_Agrtu-NOS), and a left border sequence (LB) of A. tumefaciens used for T-DNA transcription. The transgene insert in chromosome B07 of NUBJ1207 contains a right border sequence (RB) of A. tumefaciens used for T-DNA transcription, a terminator of Linum usitatissimum conlinin2 (TER_Linus-Cnl2), M. psirha Δ6 desaturase, and a truncated promoter of L. usitatissimum conlinin2 (PRO_Linus-Cnl2) (without 5’ 228 bp).The transgene insert in chromosome A08 of NUBJ1207 contains RB, TER_Linus-Cnl2, M. psirilla Δ6 desaturase, PRO_Linus-Cnl2, Pro_Arath-FAE1, P. cordata Δ5 elongase, Ter_Glyma-lectin, the promoter of B. napus napin (PRO_Brana-Fp1), P. satina Δ5 desaturase, TER_Agrtu-NOS, the terminator of L. usitatissimum conlinin1 (TER_Linus-Cnl1), P. pastoris Δ15 / ω3-desaturase, the promoter of L. usitatissimum conlinin1 (PRO_Linus-Cnl1), PRO_Linus-Cnl2, and 21 base pairs of P. satina Δ4-desaturase. (Note that the expression cassette described above reflects a vector design in which the genes are not arranged in the same 5’-3’ orientation). Further, with respect to the transgene insert in A08, the truncated Δ4-desaturase sequence encodes only the first 7 N-terminal amino acid residues of the enzyme and is not expected to have Δ4-desaturase activity, which is consistent with the fatty acid content observed in NUBJ1207 seed oil.

[0027] Three transgene inserts in line NUBJ1207 provide most of the genes from binary vector modB, excluding the Δ12-desaturase gene and the Δ4-desaturase gene. However, B. juncea contains endogenous Δ12-desaturase and Δ15 / ω3-desaturase genes, such that Δ12-unsaturation activity is supplemented by the endogenous genes. Additionally, line NUBJ1207 contains two copies of the Δ6-desaturase transgene. These transgene inserts convert oleic acid (OA) to DPA but do not further convert DPA to DHA due to the lack of Δ4-desaturase activity.

[0028] Accordingly, one embodiment provides seeds of the Brassica juncea line designated NUBJ1207, a representative sample of which has been deposited under ATCC Accession No. PTA-125954, wherein said seeds contain within their genome a first transgene locus located on chromosome 1 and containing P. cordata Δ6-elongase and phosphinothricin acetyltransferase, a second transgene locus located on chromosome 2 and containing M. psila Δ6-desaturase, and a third transgene locus located on chromosome 3 and containing a non-functional portion of M. psila Δ6-desaturase, P. cordata Δ5-elongase, P. salina Δ5-desaturase, Pichia pastoris Δ15 / ω3-desaturase, and P. salina Δ4-desaturase.

[0029] Since the initial transformation, the progeny have been self-pollinated and the plant breeding pedigree system has subsequently been used to develop line NUBJ1207 of this embodiment. Some of the criteria used for selection in various generations include DPA content (% of total fatty acids in the seed), vigor, fertility, standability, disease resistance, oil content, and maturity. NUBJ1207 is thus a genetically stable and homogeneous transgenic Brassica, and no off-type plants have been shown during evaluation. The NUBJ1207 line exhibits genetic stability and homogeneity as described in the phenotypic description information. It has been self-pollinated over a sufficient number of generations while paying careful attention to homogeneity with respect to plant type. This line has been increased while continuously observing for homogeneity.

[0030] A major phenotypic characteristic of NUBJ1207 is the production of a significant amount of DPA in its seeds. As shown herein, bulk seeds of NUBJ1207 generally contain about 10% DPA or more (% of total fatty acids in the seed).

[0031] At least one embodiment relates to a method for producing a DPA Brassica plant by crossing a first parent plant with a second parent plant, where the first plant or the second plant is a plant from the Brassica juncea line NUBJ1207. In at least one embodiment, both the first parent plant and the second parent plant may be from the line NUBJ1207. Any breeding method using the line NUBJ1207 is contemplated, examples of which include selfing, backcrossing, hybrid breeding, and crossing into a population. Any plant produced using the B. juncea line NUBJ1207 as a parent is within the scope of this embodiment. For example, the B. juncea line NUBJ1207 may be crossed with a canola-quality low-glucosinolate B. juncea line to provide a low-glucosinolate DPA B. juncea.

[0032] The additional methods of the present embodiment include the introduction of an expression vector to be introduced into plant tissue using a gene direct introduction method. Examples of such methods include those using microprojectile-mediated delivery, DNA injection, electroporation, or Agrobacterium-mediated transformation. In some embodiments, the transgenic variant of NUBJ1207 may contain at least one additional transgene, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional transgenes (in addition to those present in NUBJ1207). Accordingly, the present invention also relates to super-transformed variants of the claimed line NUBJ1207, or variants obtained by breeding NUBJ1207 with one or more other transgenic Brassicas. Such transgenes are expressed under the control of regulatory sequences well known in the art (e.g., promoters, enhancers, intervening sequences, terminators). In one embodiment, the desired traits may be one or more of herbicide tolerance, pest tolerance, disease resistance, modified seed yield, modified oil percentage, modified protein percentage, modified lodging resistance, or modified fatty acid or carbohydrate metabolism. The specific gene may be any gene well known in the art or listed herein. Examples include marker genes, polynucleotides (nucleic acid molecules) conferring resistance or tolerance to imidazolinones, sulfonylureas, glyphosate, glufosinate, 2,4-D, dicamba, L-phosphinothricin, triazine, hydroxyphenylpyruvate, dioxygenase inhibitors, protoporphyrinogen oxidase inhibitors, phenoxy propionic acids, cyclohexanones, or benzonitriles, polynucleotides encoding Bacillus thuringiensis, polynucleotides encoding phytase, polynucleotides encoding 1-acyl-glycerol-3-phosphate acyltransferase (LPAAT), galactinol synthase, or raffinose synthase, or polynucleotides conferring resistance to blackleg, white rust, or other common canola diseases, but are not limited thereto.Such transgenes and related techniques are well known in the art. See, for example, U.S. Patent Nos. 9,718,759 and 8,143,488. Alternatively, the DPA traits of such transgenic embodiments may be obtained by breeding canola containing at least one transgene with NUBJ1207 as exactly described. For example, the DPA yield may be increased by introgressing NUBJ1207 with a transgenic B. napus A02 chromosome as described in U.S. Patent No. 10,563,218. The traits for producing DHA as well as DPA may be obtained, for example, by introgressing at least one complete modB insert (one locus containing two modB constructs), such as a transgenic A05 chromosome, into NUBJ1207 as described in U.S. Patent No. 10,570,405.

[0033] Accordingly, a transgene or other phenotypic trait can be introduced into NUBJ1207, or the DPA production trait of NUBJ1207 can be introduced into other Brassica or canola lines, which are performed using conventional breeding techniques such as gene transfer or backcrossing well known in the art. Examples of canola plant breeding techniques that can be utilized in producing the progeny of NUBJ1207 include, for example, recurrent selection, bulk selection, mass selection, mutation breeding, backcrossing, pedigree breeding, tissue culture, open pollination breeding, restriction fragment length polymorphism enhanced selection, genetic marker enhanced selection, doubled haploid production. Combinations of such techniques are often used. The development of Brassica subspecies in a plant breeding program typically requires the development and evaluation of homozygous subspecies. See, for example, US2018 / 0016591, U.S. Patent No. 8,143,488, Downey et al., Rapeseed & Mustard, at 437 in PRINCIPLES OF CULTIVAR DEVEL. (Fehr (ed.), Macmillan & Co., NY, 1987), Thompson, Breeding winter oilseed rape Brassica napus, 7 Adv. Appl. Biol. 1-104 (1983), Ward et al., Oilseed Rape (Farming Press Ltd., Wharefedale Road, Ipswich, Suffolk, 1985).

[0034] In another aspect of this embodiment, NUBJ1207 and its progeny can be identified not only by their phenotypic traits (i.e., the production of DPA), but also by the identification of their genotypes. As described in Example 3 of this specification, NUBJ1207 not only has a unique gene profile brought about by its transgene, but also contains unique DNA sequences at the junctions created upon insertion of the gene cassette into the B. juncea genome. More specifically, a part of the gene construct GA7-modB is present at three loci, each of which is on a different chromosome (chromosomes A08, A06, and B07), and each creates six unique junctions with unique DNA sequences.

[0035] The marker profile of NUBJ1207 can be recognized using techniques such as RFLP, random amplified polymorphic DNA (RAPD), arbitrarily primed polymerase chain reaction (AP-PCR), DNA amplified fingerprinting (DAF), sequence characterized amplified region (SCAR), amplified fragment length polymorphism (AFLP), SSR, also called microsatellite, or SNP. See, for example, Glick et al., METHODS IN PLANT MOLEC. BIOL. & BIOTECHNOL. (CRC Press, Boca Raton, 1993). An exemplary assay for the identification of NUBJ1207 junctions is the Kompetitive Allele Specific PCR (KASP) assay, the method of which is illustrated in US2018 / 0016591.

[0036] Currently, canola is recognized as an increasingly important oilseed crop and as a source of meal in many parts of the world. The DPA juncea line NUBJ1207 or its progeny can be used in the production of oil, meal, or other food or edible supplies, including nutritional supplements or pharmaceuticals, according to known techniques. Advantageously, the oil obtained from DPA juncea NUBJ1207 has a low erucic acid content (less than 1%). Glucosinolates may be found in the meal portion of the processed seeds and can be minimized by methods known in the art.

[0037] In accordance with the Budapest Treaty, the applicant has deposited the seeds of DPA B. juncea NUBJ1207 with the American Type Culture Collection (ATCCR), located at 0801 University Blvd., Manassas, Va., 20110-2209 U.S.A., under accession number PTA-125954. The applicant has met the requirements of 37 C.F.R. §§ 1.801-1.809. The applicant does not have the authority to waive any restrictions imposed by laws regarding the movement or transportation of biological materials in commerce. The applicant does not waive any rights granted under patent law or plant breeder's rights.

[0038] Accordingly, one aspect of the present embodiment provides seeds of the Brassica juncea line designated NUBJ1207 (DPA juncea NUBJ1207), and representative samples of such seeds are deposited under ATCC accession number PTA-125954. At least one embodiment provides a Brassica plant or a part thereof, which contains the genome of the seeds of DPA juncea NUBJ1207 generated by growing the seeds of DPA juncea NUBJ1207, and examples thereof include seeds, pollen, cotyledons, half-cotyledons, tissue cultures of regenerable cells, or ovules. The tissue culture or regenerable cells according to one embodiment can regenerate a plant that can express all the morphological and physiological characteristics of DPA juncea NUBJ1207, and may constitute leaves, pollen, embryos, roots, root tips, sheaths, flowers, ovules, and petioles. Another embodiment provides a Brassica plant or a part thereof that has all the physiological and morphological characteristics of DPA juncea NUBJ1207. Another embodiment provides cells of the B. juncea line NUBJ1207, and representative samples of such seeds are deposited under ATCC accession number PTA-125954.

[0039] Another aspect of the present embodiment provides a method for producing Brassica seeds, the method including the steps of growing and cultivating DPA juncea NUBJ1207 to produce and harvest the seeds, and representative samples of such seeds are deposited under ATCC accession number PTA-125954. The step of growing or cultivating can be carried out in a greenhouse, a tent, or a field. The step of growing or cultivating can be carried out on a commercial scale. At least one embodiment provides a composition containing oil, meal, or meal protein obtained by practicing this method. The oil may be crude oil, refined oil, or concentrated oil of one or more fatty acids.

[0040] Another aspect of this embodiment provides a method for producing oil or meal, the method comprising obtaining seeds of DPA juncea NUBJ1207, or related progeny thereof, and processing the seeds to obtain oil or meal. At least one embodiment provides a composition comprising oil from seeds of DPA juncea NUBJ1207, or related progeny thereof. The oil may be crude, refined, or concentrated oil of one or more fatty acids. At least one embodiment provides a composition comprising meal from seeds of DPA juncea NUBJ1207, or related progeny thereof.

[0041] Another aspect of this embodiment provides a Brassica plant or a part thereof (e.g., progeny) derived from DPA juncea NUBJ1207, wherein the plant or part thereof is transformed such that its genetic material comprises one or more transgenes operably linked to one or more regulatory elements in addition to the transgene present in the Brassica juncea line NUBJ1207. Thus, at least one embodiment provides a method for producing a Brassica plant whose genetic material comprises at least one transgene in addition to the transgene of line NUBJ1207, the method comprising crossing a NUBJ1207 plant or a plant derived therefrom with a second plant of another Brassica line, whereby the genetic material of the progeny resulting from the cross expresses at least one additional transgene. Alternatively, at least one embodiment provides a method for producing a Brassica plant whose genetic material comprises at least one transgene in addition to the transgene of line NUBJ1207, the method comprising introducing at least one additional transgene into NUBJ1207. In such methods, the genetic material of the progeny resulting from the cross or transformation expresses desirable additional traits. The additional transgene may confer herbicide tolerance, insect pest tolerance, bacterial disease tolerance, fungal disease tolerance, viral disease tolerance, or sterility.

[0042] Another embodiment provides a method for generating a canola plant derived from the Brassica juncea line NUBJ1207, the method comprising: (a) crossing the B. juncea line NUBJ1207 with a second canola plant to produce progeny canola seeds; and (b) growing the progeny canola seeds under plant growth conditions to produce a canola plant derived from the Brassica line NUBJ1207. This embodiment further includes: (c) an additional step of crossing a canola plant derived from NUBJ1207 with itself or with another canola plant to produce additional canola progeny seeds derived from NUBJ1207; (d) an additional step of growing the progeny canola seeds of step (c) under plant growth conditions to produce an additional canola plant derived from NUBJ1207; and (e) repeating the crossing step of (c) and the growing step of (d) from zero (0) to ten (10) times to further produce a canola plant derived from NUBJ1207, wherein the canola plant derived from NUBJ1207 expresses at least two NUBJ1207 traits selected from DPA products and at least one additional agriculturally economically desirable trait. Related embodiments provide a canola plant or a part thereof derived from NUBJ1207 produced by such a method.

[0043] Another embodiment provides a method for generating Brassica seeds, the method including crossing a first parent Brassica plant with a second parent Brassica plant and harvesting the resulting first-generation Brassica seeds, wherein the first parent Brassica plant or the second parent Brassica plant is a Brassica line NUBJ1207 plant. Specifically, the first parent Brassica plant is different from the second parent Brassica plant, and the resulting seeds are first-generation (F1) hybrid Brassica seeds. Related embodiments provide further steps of backcrossing a selected F1 hybrid plant with line NUBJ1207 or a different Brassica plant having desirable traits to generate backcross progeny seeds, growing the resulting backcross progeny seeds and selecting backcross progeny plants having the desirable traits, and repeating the backcrossing and growing steps over 1 to 10 generations on the selected backcross progeny plants to generate progeny Brassica plants derived from Brassica juncea line NUBJ1207. In related embodiments, progeny derived from Brassica juncea line NUBJ1207 can be obtained by self-pollinating (selfing) the F1 progeny.

[0044] Another embodiment provides a method for obtaining an inbred line of Brassica, the method including: (a) planting a collection of seeds including hybrid seeds, one of whose parents is Brassica NUBJ1207, the collection further including seeds of Brassica NUBJ1207; (b) growing canola plants from the collection of seeds; (c) identifying inbred plants from the inbred line; (d) selecting the inbred plants; and (e) controlling pollination in a manner that maintains the homozygosity of the inbred plants. Seeds of the inbred Brassica line NUBJ1207 are deposited under ATCC Accession No. PTA-125954. In some embodiments of this method, the inbred line of Brassica may be B. juncea or B. napus.

[0045] At least one embodiment provides a method for generating doubled monoploid sub-species, the method comprising: (a) isolating buds of F1 plants provided herein; (b) dissecting monoploid anther pollens; (c) placing the monoploid anther pollens in a culture; (d) inducing the anther pollens to differentiate into embryos and then into further smaller plantlets; (e) identifying whether the further smaller plantlets contain a diploid chromosome number, wherein the diploid chromosome number is generated through chromosome doubling; and, when the further smaller plantlets contain a diploid chromosome number, continuing to grow the smaller plantlets.

[0046] A further aspect of this embodiment provides novel nucleic acid molecules (e.g., DNA molecules) embodied by the combination of transgene inserts and the B. juncea genome, and cells or compositions containing such nucleic acids. Exemplary embodiments provide DNA molecules comprising one, two, or three DNA molecules selected from: (a) a DNA molecule comprising the sequence provided as SEQ ID NO: 1 and the sequence provided as SEQ ID NO: 2; (b) a DNA molecule comprising the sequence provided as SEQ ID NO: 3 and the sequence provided as SEQ ID NO: 4; or (c) a DNA molecule comprising the sequence provided as SEQ ID NO: 5 and the sequence provided as SEQ ID NO: 6. Exemplary compositions according to this aspect may be seed meals containing such DNA molecules. At least one embodiment provides plants (such as Brassica plants), seeds, plant parts, seed meals, etc., described herein that contain such DNA molecules.

[0047] The related embodiments provide a cell comprising one, two, or three nucleic acid molecules selected from: (a) a DNA molecule comprising the sequence provided as SEQ ID NO:1 and the sequence provided as SEQ ID NO:2; (b) a DNA molecule comprising the sequence provided as SEQ ID NO:3 and the sequence provided as SEQ ID NO:4; (c) a DNA molecule comprising the sequence provided as SEQ ID NO:5 and the sequence provided as SEQ ID NO:6. The cell may be a B. juncea cell. The cell may be present in a plant, seed, or part of a plant as described herein. The cell may be present in meal (i.e., seed meal).

[0048] Another aspect provides a method of using the DNA molecules of SEQ ID NOs:1-6, or a portion thereof. Such use includes, for example, identifying the presence of at least one of the locus characteristics of NUBJ1207. Certain embodiments provide a method of detecting a DNA molecule, such as amplification of a DNA molecule comprising a sequence of at least 20 contiguous nucleotides from one of SEQ ID NOs:1-6. In certain embodiments, the method may be a KASP assay. Related embodiments provide a single-stranded portion of any one of the nucleic acids of SEQ ID NOs:1-6, such as a primer or probe.

[0049] Another aspect of this embodiment provides a method of detecting the presence of event NUBJ1207 in a sample comprising plant DNA, wherein the method comprises contacting the sample with at least one primer that binds to at least one transgene flanking region of the Brassica genome as shown in SEQ ID NOs:1-6. The method may utilize a KASP™ genotyping assay. Related embodiments provide a plant, plant material, or plant-derived material identified by such a method. Another related embodiment provides a kit comprising components for performing such a method.

[0050] At least one embodiment provides a method for detecting the presence of event NUBJ1207 in a sample containing plant DNA, the method comprising: (a) contacting the sample with a first primer that binds to an adjacent junction region of the Brassica genome of NUBJ1207 and a second primer that binds to the transgene of NUBJ1207; (b) subjecting the sample to a polymerase chain reaction; and (c) characterizing the amplicon produced between the primers.

[0051] Another aspect of this embodiment provides genomic DNA of the inbred B. juncea line NUBJ1207, and representative seeds of the inbred line have been deposited under ATCC Accession No. PTA-125954. Related embodiments provide plant cells comprising: (a) DNA having a sequence shown in at least one of SEQ ID NO: 1 to SEQ ID NO: 6; (b) DNA having at least 95% identity with at least one of SEQ ID NO: 1 to SEQ ID NO: 6; or (c) a complement of (a) or (b).

[0052] Another aspect of this embodiment provides seeds of the B. juncea line NUBJ1207, or plants derived therefrom (i.e., progeny or related progeny), comprising about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, or about 20% DPA (as % of total fatty acids in the seed oil). Another embodiment provides a collection of seeds, or plants derived therefrom, comprising at least 95% of the seeds of the B. juncea line NUBJ1207, wherein the seeds comprise about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, or about 20% DPA. Certain embodiments provide seed oil obtained from such seeds or collections of seeds.

[0053] A further aspect provides a method for obtaining DPA-containing Brassica grains, comprising: (a) introducing the DPA trait of NUBJ1207 into an elite Brassica line that is male sterile; (b) introducing the DPA trait of NUBJ1207 into a second elite Brassica line that is fertile; (c) crossing the two lines (a) and (b) to obtain hybrid progeny; (d) cultivating the seeds of the hybrid progeny; and (e) harvesting the grains produced by the cultivated hybrid progeny.

Example

[0054] Example 1. Cultivation attempt (North America) Seeds obtained from the B line derived from the elite event of NUBJ1207 had the following fatty acid profile as determined by gas chromatography (Table 2).

[0055]

Table 2

[0056] The seeds were sown in the spring in Zone 7 of North America. During cultivation, specific agronomic characteristics were monitored approximately weekly as shown in Table 3 below.

[0057]

Table 3

[0058] Example 2. Cultivation attempt (Australia) Four lines of B. juncea for DPA production containing NUBJ1207 were field-trialed in separate tents in Australia, and the fatty acid content of the mature seeds (pooled bulk) was analyzed by gas chromatography (alternatively, seed fatty acids can also be measured using high-resolution NMR). Fatty acid contents where the existing fatty acids exceed 1% (of the total fatty acids %) are shown in Table 4 below. The data show consistency between the generations from bulk plants and the generations from single plants.

[0059]

Table 4

[0060] DPA B. juncea NUBJ1207 was grown in the field in southeastern Australia during the winter of 2017. Seeds were sown in May, and the resulting plants began to flower 139 days after sowing, and the seeds were harvested 218 days after sowing.

[0061] Pooled seed data from single plants from siblings of tent-grown NUBJ1207 are shown in Table 5 below. The data show consistency among siblings of the T5 generation.

[0062]

Table 5

[0063] In further experiments, seed sheaths were harvested from 10 individual tent-grown plants at various stages of maturity and plant position, and the seeds were analyzed. The following data, namely Table 6, show consistency among sheaths with different maturity and position in the plants.

[0064]

Table 6

[0065] Example 4. Unique Adjacent Regions The methods and kits of this specification are useful for identifying the presence of transgenic plant material, particularly plant material containing a transgene in NUBJ1207, as well as transgenic Brassica plants, plant material, and seeds containing such events in biological samples. The elite event NUBJ1207 described herein can be identified by genotype, which can be characterized through a genetic marker profile that can identify plants of the same cultivar or related cultivars, or can be used to determine or confirm a pedigree. The genetic marker profile can be obtained by techniques such as restriction fragment length polymorphism (RFLP), random amplified polymorphic DNA (RAPD), arbitrarily primed polymerase chain reaction (AP-PCR), DNA amplification fingerprinting (DAF), sequence characterized amplified region (SCAR), amplified fragment length polymorphism (AFLP), simple sequence repeats (SSR) (also called microsatellites), and single nucleotide polymorphisms (SNP).

[0066] For example, the elite event NUBJ1207 described herein can be identified by generating a genetic map from a sample of plant material. The genetic map can be generated by conventional RFLP, polymerase chain reaction (PCR) analysis, or SSRs that identify the approximate chromosomal location of an integrated DNA molecule encoding a heterologous protein. See Glick & Thompson, METHODS IN PLANT MOLEC. BIOL. & BIOTECHNOL. 269 (CRC Press, Boca Raton, FL, 1993). Map information regarding chromosomal location is useful for the owner protection of the transgenic plant in question. For example, the map of the integration region can be compared with a similar map of a suspect plant to determine whether the latter has a common lineage with the target plant. Examples of map comparisons can include hybridization, RFLP, PCR, SSR, and sequencing, all of which are prior art.

[0067] DPA Junscea NUBJ1207 is an inbred line from an elite event that can be characterized and identified by unique DNA sequences at the junction of the parental plant genome and the inserted transgene construct. More specifically, each of chromosomes A08, A06, and B07 contains 5' and 3' junctions unique to DPA Junscea.

[0068] For example, the DNA sequence of the 5' junction of chromosome A08 contains 100 nucleotides of B. Junscea (italicized) and 100 nucleotides of the transgene (non-italicized).

[0069]

Chemical formula

[0070] For example, the DNA sequence of the 3' junction of chromosome A08 contains 100 nucleotides of the transgene (non-italicized) and 100 nucleotides of B. Junscea (italicized).

[0071]

Chemical formula

[0072] For example, the DNA sequence of the 5' junction of chromosome A06 contains 100 nucleotides of B. Junscea (italicized) and 100 nucleotides of the transgene (non-italicized).

[0073]

Chemical formula

[0074] For example, the DNA sequence of the 3' junction of chromosome A06 contains 100 nucleotides of the transgene (non-italicized) and 100 nucleotides of B. Junscea (italicized).

[0075]

Chemical formula

[0076] For example, the DNA sequence of the 5'-junction of chromosome B07 contains 100 nucleotides of B. juncea (italic) and 100 nucleotides of the transgene (non-italic).

[0077]

Chem.

[0078] For example, the DNA sequence of the 3'-junction of chromosome B07 contains 100 nucleotides of the transgene (non-italic) and 100 nucleotides of B. juncea (italic).

[0079]

Chem.

[0080] For the identification of DPA juncea NUBJ1207, the primers for identifying the junction can be of any suitable length derived from the larger junction sequence. The length of the junction sequence used to identify DPA Juncea NUBJ1207 may be any suitable length that provides identification of the unique junction, for example, at least 10, or at least 15, or 20 consecutive nucleotides from the junction of the transgene and B. juncea DNA.

[0081] Although the foregoing embodiments have been described in considerable detail for purposes of clarity and understanding, it will be apparent to those skilled in the art that certain changes and modifications, such as single gene modifications and mutations, somaclonal variants, mutant individuals selected from large populations of inbred lines of plants, etc., may be practiced within the scope of the invention as limited only by the appended claims.

Claims

**Claim 1** Seeds of the Brassica juncea line designated NUBJ1207, said seeds having been deposited under ATCC accession number PTA-125954, wherein DPA juncea NUBJ1207 has within its genome, a transgene locus on chromosome A08 comprising a 5' flanking sequence of a chromosome A08 transgene insert of the nucleotide sequence of SEQ ID NO: 1 or a nucleotide sequence complementary to SEQ ID NO: 1, and a 3' flanking sequence of a chromosome A08 transgene insert of the nucleotide sequence of SEQ ID NO: 2 or a nucleotide sequence complementary to SEQ ID NO: 2, a transgene locus on chromosome A06 comprising a 5' flanking sequence of a chromosome A06 transgene insert of the nucleotide sequence of SEQ ID NO: 3 or a nucleotide sequence complementary to SEQ ID NO: 3, and a 3' flanking sequence of a chromosome A06 transgene insert of the nucleotide sequence of SEQ ID NO: 4 or a nucleotide sequence complementary to SEQ ID NO: 4, a transgene locus on chromosome B07 comprising a 5' flanking sequence of a chromosome B07 transgene insert of the nucleotide sequence of SEQ ID NO: 5 or a nucleotide sequence complementary to SEQ ID NO: 5, and a 3' flanking sequence of a chromosome B07 transgene insert of the nucleotide sequence of SEQ ID NO: 6 or a nucleotide sequence complementary to SEQ ID NO: 6, comprising, wherein the transgene locus on chromosome A08 comprises transgenes of M. psirra Δ6 desaturase, P. cordata Δ5 elongase, P. marina Δ5 desaturase, and P. pastoris Δ15 / ω3-desaturase; the transgene locus on chromosome A06 comprises a transgene of P. cordata Δ6-elongase; and the transgene locus on chromosome B07 comprises a transgene of M. psirra Δ6 desaturase, seeds, which do not contain a transgene for Δ12 desaturase within their genome. **Claim 2** A Brassica plant or a part thereof produced by growing the seeds of claim 1. **Claim 3** The Brassica plant or a part thereof according to claim 2, wherein the part is at least one of seeds, leaves, pollen, embryos, roots, root tips, sheaths, flowers, ovules, stems, cells, protoplasts, or tissue cultures. **Claim 4** A method for producing omega-3 docosapentaenoic acid (DPA) in the seeds of the Brassica plant according to claim 2, said method comprising growing the plant or a progeny thereof that produces DPA and has within its genome, The 5'-flanking junction sequence of the chromosomal A08 transgene insert of SEQ ID NO: 1 or a nucleotide sequence complementary to SEQ ID NO: 1, and the 3'-flanking junction sequence of the chromosomal A08 transgene insert of SEQ ID NO: 2 or a nucleotide sequence complementary to SEQ ID NO: 2, a gene introduction locus on chromosome A08 comprising: The 5'-flanking junction sequence of the chromosomal A06 transgene insert of SEQ ID NO: 3 or a nucleotide sequence complementary to SEQ ID NO: 3, and the 3'-flanking junction sequence of the chromosomal A06 transgene insert of SEQ ID NO: 4 or a nucleotide sequence complementary to SEQ ID NO: 4, a gene introduction locus on chromosome A06 comprising: The 5'-flanking junction sequence of the chromosomal B07 transgene insert of SEQ ID NO: 5 or a nucleotide sequence complementary to SEQ ID NO: 5, and the 3'-flanking junction sequence of the chromosomal B07 transgene insert of SEQ ID NO: 6 or a nucleotide sequence complementary to SEQ ID NO: 6, a gene introduction locus on chromosome B07 comprising: comprising wherein the gene introduction locus on chromosome A08 comprises the transgenes of M. psirra Δ6 desaturase, P. cordata Δ5 elongase, P. salina Δ5 desaturase, and P. pastoris Δ15 / ω3-desaturase; the gene introduction locus on chromosome A06 comprises the transgene of P. cordata Δ6-elongase; the gene introduction locus on chromosome B07 comprises the transgene of M. psirra Δ6 desaturase, and a progeny that does not contain the transgene of Δ12 desaturase in its genome. A method comprising the step of cultivating. **Claim 5** A method for producing Brassica seeds, the method being the Brassica plant or its progeny according to claim 2 for producing seeds, producing DPA, and within its genome, The 5'-flanking junction sequence of the chromosomal A08 transgene insert of SEQ ID NO: 1 or a nucleotide sequence complementary to SEQ ID NO: 1, and the 3'-flanking junction sequence of the chromosomal A08 transgene insert of SEQ ID NO: 2 or a nucleotide sequence complementary to SEQ ID NO: 2, a gene introduction locus on chromosome A08 comprising: The 5'-flanking junction sequence of the chromosomal A06 transgene insert of SEQ ID NO: 3 or a nucleotide sequence complementary to SEQ ID NO: 3, and the 3'-flanking junction sequence of the chromosomal A06 transgene insert of SEQ ID NO: 4 or a nucleotide sequence complementary to SEQ ID NO: 4, a gene introduction locus on chromosome A06 comprising: The 5'-flanking junction sequence of the chromosomal B07 transgene insert of the nucleotide sequence of SEQ ID NO: 5 or a nucleotide sequence complementary to SEQ ID NO: 5, and the 3'-flanking junction sequence of the chromosomal B07 transgene insert of the nucleotide sequence of SEQ ID NO: 6 or a nucleotide sequence complementary to SEQ ID NO: 6, and a transgenic locus on chromosome B07 comprising the same, comprising, wherein the transgenic locus on chromosome A08 comprises transgenes of M. psyllae Δ6 desaturase, P. cordata Δ5 elongase, P. salina Δ5 desaturase, and P. pastoris Δ15 / ω3-desaturase; the transgenic locus on chromosome A06 comprises the transgene of P. cordata Δ6-elongase; the transgenic locus on chromosome B07 comprises the transgene of M. psyllae Δ6 desaturase, a method comprising the steps of growing a progeny that does not contain the transgene of Δ12 desaturase in its genome, and harvesting the seeds. **Claim 6** The seeds of a Brassica plant harvested as described in claim 5. **Claim 7** A Brassica plant or its progeny according to claim 2, which produces DPA and in its genome, the 5'-flanking junction sequence of the chromosomal A08 transgene insert of the nucleotide sequence of SEQ ID NO: 1 or a nucleotide sequence complementary to SEQ ID NO: 1, and the 3'-flanking junction sequence of the chromosomal A08 transgene insert of the nucleotide sequence of SEQ ID NO: 2 or a nucleotide sequence complementary to SEQ ID NO: 2, and a transgenic locus on chromosome A08 comprising the same, the 5'-flanking junction sequence of the chromosomal A06 transgene insert of the nucleotide sequence of SEQ ID NO: 3 or a nucleotide sequence complementary to SEQ ID NO: 3, and the 3'-flanking junction sequence of the chromosomal A06 transgene insert of the nucleotide sequence of SEQ ID NO: 4 or a nucleotide sequence complementary to SEQ ID NO: 4, and a transgenic locus on chromosome A06 comprising the same, the 5'-flanking junction sequence of the chromosomal B07 transgene insert of the nucleotide sequence of SEQ ID NO: 5 or a nucleotide sequence complementary to SEQ ID NO: 5, and the 3'-flanking junction sequence of the chromosomal B07 transgene insert of the nucleotide sequence of SEQ ID NO: 6 or a nucleotide sequence complementary to SEQ ID NO: 6, and a transgenic locus on chromosome B07 comprising the same, comprising, Here, the transgene locus on chromosome A08 contains the transgenes of M. psyllae Δ6 desaturase, P. cordata Δ5 elongase, P. salina Δ5 desaturase, and P. pastoris Δ15 / ω3-desaturase; the transgene locus on chromosome A06 contains the transgene of P. cordata Δ6-elongase; and the transgene locus on chromosome B07 contains the transgene of M. psyllae Δ6 desaturase. A method for producing oil, comprising the steps of obtaining progeny seeds that do not contain the transgene of Δ12 desaturase in their genome, and treating said seeds to obtain oil. **Claim 8** A Brassica plant or its progeny according to claim 2, which produces DPA and, in its genome, a transgene locus on chromosome A08 comprising a 5' flanking sequence of a chromosome A08 transgene insert having the nucleotide sequence of SEQ ID NO: 1 or a nucleotide sequence complementary to SEQ ID NO: 1, and a 3' flanking sequence of a chromosome A08 transgene insert having the nucleotide sequence of SEQ ID NO: 2 or a nucleotide sequence complementary to SEQ ID NO: 2; a transgene locus on chromosome A06 comprising a 5' flanking sequence of a chromosome A06 transgene insert having the nucleotide sequence of SEQ ID NO: 3 or a nucleotide sequence complementary to SEQ ID NO: 3, and a 3' flanking sequence of a chromosome A06 transgene insert having the nucleotide sequence of SEQ ID NO: 4 or a nucleotide sequence complementary to SEQ ID NO: 4; a transgene locus on chromosome B07 comprising a 5' flanking sequence of a chromosome B07 transgene insert having the nucleotide sequence of SEQ ID NO: 5 or a nucleotide sequence complementary to SEQ ID NO: 5, and a 3' flanking sequence of a chromosome B07 transgene insert having the nucleotide sequence of SEQ ID NO: 6 or a nucleotide sequence complementary to SEQ ID NO: 6; and comprising Here, the transgenic locus on chromosome A08 contains the introduced genes of M. psyllae Δ6 desaturase, P. cordata Δ5 elongase, P. salina Δ5 desaturase, and P. pastoris Δ15 / ω3-desaturase; the transgenic locus on chromosome A06 contains the introduced gene of P. cordata Δ6-elongase; the transgenic locus on chromosome B07 contains the introduced gene of M. psyllae Δ6 desaturase. A collection of seeds containing at least 95% of the seeds of the progeny that do not contain the introduced gene of Δ12 desaturase in their genome, wherein the seeds contain about 5% to 20% of omega-3 docosapentaenoic acid (DPA) (as the weight percentage of the total fatty acids in the seed oil).

9. Seed oil obtained from the collection of seeds according to Claim 8.

10. Meal obtained from the collection of seeds according to Claim 8.

11. A Brassica plant or its progeny according to Claim 2, containing fatty acids in an esterified form, producing DPA, and having in its genome a transgenic locus on chromosome A08 comprising a 5' flanking sequence of the chromosome A08 transgenic insert of the nucleotide sequence of SEQ ID NO: 1 or a nucleotide sequence complementary to SEQ ID NO: 1, and a 3' flanking sequence of the chromosome A08 transgenic insert of the nucleotide sequence of SEQ ID NO: 2 or a nucleotide sequence complementary to SEQ ID NO: 2 a transgenic locus on chromosome A06 comprising a 5' flanking sequence of the chromosome A06 transgenic insert of the nucleotide sequence of SEQ ID NO: 3 or a nucleotide sequence complementary to SEQ ID NO: 3, and a 3' flanking sequence of the chromosome A06 transgenic insert of the nucleotide sequence of SEQ ID NO: 4 or a nucleotide sequence complementary to SEQ ID NO: 4 a transgenic locus on chromosome B07 comprising a 5' flanking sequence of the chromosome B07 transgenic insert of the nucleotide sequence of SEQ ID NO: 5 or a nucleotide sequence complementary to SEQ ID NO: 5, and a 3' flanking sequence of the chromosome B07 transgenic insert of the nucleotide sequence of SEQ ID NO: 6 or a nucleotide sequence complementary to SEQ ID NO: 6 and including Here, the transgenic locus on chromosome A08 contains the transgenes of M. psyllae Δ6 desaturase, P. cordata Δ5 elongase, P. salina Δ5 desaturase, and P. pastoris Δ15 / ω3-desaturase; the transgenic locus on chromosome A06 contains the transgene of P. cordata Δ6-elongase; the transgenic locus on chromosome B07 contains the transgene of M. psyllae Δ6 desaturase. An extracted seed oil obtained from progeny that do not contain the transgene of Δ12 desaturase in their genome, wherein the fatty acids include palmitic acid (C16), stearic acid (C18), oleic acid (C18:1), cis-vaccenic acid (C18:1n7c), linoleic acid (C18:2n6c), α-linolenic acid (C18:3n3), stearidonic acid (C18:4n3), gondoic acid (C20:1n9c), eicosatetraenoic acid (C20:4n3), ω3-docosatrienoic acid (C22:4n3), and ω3-docosapentaenoic acid (C22:5n3). Here, in the total fatty acid content of the seed oil, the level of palmitic acid is less than 5%, the level of stearic acid is less than 4%, the level of oleic acid is 35% to 48%, the level of cis-vaccenic acid is less than 4%, the level of linoleic acid is less than 9%, the level of α-linolenic acid is 15% to 20%, the level of stearidonic acid is less than 2%, the level of gondoic acid is less than 2%, the level of eicosatetraenoic acid is less than 4.5%, the level of ω3-docosatrienoic acid is 1% to 3%, and the level of ω3-docosapentaenoic acid is 8% to 15%. An extracted seed oil. [

12. ] A method for producing a Brassica plant having at least one desirable trait in addition to the traits of the inbred Brassica line NUBJ1207 deposited under ATCC accession number PTA-125954, wherein the trait of the inbred Brassica line NUBJ1207 is the production of omega-3 docosapentaenoic acid, the method comprising crossing a plant of the inbred line NUBJ1207 with another Brassica elite event, whereby the genetic material resulting from the crossing expresses the desirable additional trait, wherein the inbred Brassica line NUBJ1207 has in its genome The 5'-flanking junction sequence of the chromosomal A08 transgene insert of the nucleotide sequence of SEQ ID NO: 1 or a nucleotide sequence complementary to SEQ ID NO: 1, and the 3'-flanking junction sequence of the chromosomal A08 transgene insert of the nucleotide sequence of SEQ ID NO: 2 or a nucleotide sequence complementary to SEQ ID NO: 2, and a transgenic locus on chromosome A08 comprising the same, The 5'-flanking junction sequence of the chromosomal A06 transgene insert of the nucleotide sequence of SEQ ID NO: 3 or a nucleotide sequence complementary to SEQ ID NO: 3, and the 3'-flanking junction sequence of the chromosomal A06 transgene insert of the nucleotide sequence of SEQ ID NO: 4 or a nucleotide sequence complementary to SEQ ID NO: 4, and a transgenic locus on chromosome A06 comprising the same, The 5'-flanking junction sequence of the chromosomal B07 transgene insert of the nucleotide sequence of SEQ ID NO: 5 or a nucleotide sequence complementary to SEQ ID NO: 5, and the 3'-flanking junction sequence of the chromosomal B07 transgene insert of the nucleotide sequence of SEQ ID NO: 6 or a nucleotide sequence complementary to SEQ ID NO: 6, and a transgenic locus on chromosome B07 comprising the same, comprising, wherein the transgenic locus on chromosome A08 comprises transgenes of M. psirra Δ6 desaturase, P. cordata Δ5 elongase, P. marina Δ5 desaturase, and P. pastoris Δ15 / ω3-desaturase; the transgenic locus on chromosome A06 comprises the transgene of P. cordata Δ6-elongase; and the transgenic locus on chromosome B07 comprises the transgene of M. psirra Δ6 desaturase, the inbred Brassica line NUBJ1207 does not contain the transgene of Δ12 desaturase in its genome, the additional trait is at least one of increased DPA production, canola traits, herbicide tolerance, insect pest tolerance, bacterial disease resistance, fungal disease resistance, viral disease resistance, or sterility, method.

13. A method for producing a canola plant derived from Brassica line NUBJ1207, the method comprising: (a) crossing a plant of Brassica juncea line NUBJ1207 deposited under ATCC Accession No. PTA-125954 with a second Brassica plant to produce progeny Brassica seeds; (b) growing the progeny Brassica seeds under plant growth conditions to produce a canola plant derived from the Brassica line NUBJ1207, wherein the Brassica line NUBJ1207 has in its genome, The gene integration locus on chromosome A08, comprising the 5' flanking junction sequence of the chromosome A08 transgene insert of SEQ ID NO: 1 or a nucleotide sequence complementary to SEQ ID NO: 1, and the 3' flanking junction sequence of the chromosome A08 transgene insert of SEQ ID NO: 2 or a nucleotide sequence complementary to SEQ ID NO:

2. The gene integration locus on chromosome A06, comprising the 5' flanking junction sequence of the chromosome A06 transgene insert of SEQ ID NO: 3 or a nucleotide sequence complementary to SEQ ID NO: 3, and the 3' flanking junction sequence of the chromosome A06 transgene insert of SEQ ID NO: 4 or a nucleotide sequence complementary to SEQ ID NO:

4. The gene integration locus on chromosome B07, comprising the 5' flanking junction sequence of the chromosome B07 transgene insert of SEQ ID NO: 5 or a nucleotide sequence complementary to SEQ ID NO: 5, and the 3' flanking junction sequence of the chromosome B07 transgene insert of SEQ ID NO: 6 or a nucleotide sequence complementary to SEQ ID NO:

6. comprising wherein the gene integration locus on chromosome A08 contains the transgenes of M. psirilla Δ6 desaturase, P. cordata Δ5 elongase, P. salina Δ5 desaturase, and P. pastoris Δ15 / ω3-desaturase; the gene integration locus on chromosome A06 contains the transgene of P. cordata Δ6-elongase; and the gene integration locus on chromosome B07 contains the transgene of M. psirilla Δ6 desaturase. The Brassica line NUBJ1207 does not contain the transgene of Δ12 desaturase in its genome. Method. [

14. ] A method for generating Brassica hybrid progeny useful for the production of omega-3 docosapentaenoic acid (DPA), the method comprising: (a) transferring the gene of NUBJ1207 deposited under ATCC accession number PTA-125954 into an elite Brassica line that is sterile; (b) transferring the gene of NUBJ1207 into a second elite Brassica line that is fertile; (c) crossing the two lines (a) and (b) to obtain hybrid progeny; and (d) cultivating the seeds of the hybrid progeny. wherein the Brassica line NUBJ1207 has in its genome The gene integration locus on chromosome A08, comprising the 5' flanking junction sequence of the chromosome A08 transgene insert of SEQ ID NO: 1 or a nucleotide sequence complementary to SEQ ID NO: 1, and the 3' flanking junction sequence of the chromosome A08 transgene insert of SEQ ID NO: 2 or a nucleotide sequence complementary to SEQ ID NO:

2. The 5'-flanking junction sequence of the chromosomal A06 transgene insert of SEQ ID NO: 3 or a nucleotide sequence complementary to SEQ ID NO: 3, and the 3'-flanking junction sequence of the chromosomal A06 transgene insert of SEQ ID NO: 4 or a nucleotide sequence complementary to SEQ ID NO: 4, a transgenic locus on chromosome A06 comprising The 5'-flanking junction sequence of the chromosomal B07 transgene insert of SEQ ID NO: 5 or a nucleotide sequence complementary to SEQ ID NO: 5, and the 3'-flanking junction sequence of the chromosomal B07 transgene insert of SEQ ID NO: 6 or a nucleotide sequence complementary to SEQ ID NO: 6, a transgenic locus on chromosome B07 comprising comprising wherein the transgenic locus on chromosome A08 comprises transgenes of M. psyllae Δ6 desaturase, P. cordata Δ5 elongase, P. salina Δ5 desaturase, and P. pastoris Δ15 / ω3-desaturase; the transgenic locus on chromosome A06 comprises the transgene of P. cordata Δ6-elongase; the transgenic locus on chromosome B07 comprises the transgene of M. psyllae Δ6 desaturase The Brassica line NUBJ1207 does not contain the transgene of Δ12 desaturase in its genome. Method. **Claim 15** The method according to claim 14, wherein the hybrid progeny comprises at least one trait selected from increased DPA production, canola oil properties, herbicide tolerance, pest tolerance, bacterial disease resistance, fungal disease resistance, viral disease resistance, or sterility.

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