PLANTS AND SEEDS OF BRASSICA CARINATA VARIETY DH-056.149

Abstract

The invention is in the field of Brassica carinata breeding (i.e. Ethiopian mustard breeding), specifically relating to Brassica carinata variety DH-056.149. The present invention relates to seeds, plants or parts thereof, cells, methods of making, and uses of this variety and its progeny.

Claims

1. A Brassica carinata variety DH-056.149, representative seed of said variety having been deposited under NCIMB accession number 43000.

2. A seed, plant, plant part or cell of Brassica carinata variety DH-056.149, representative seed of said variety having been deposited under NCIMB accession number 43000.

3. A Brassica carinata plant or plant part having the physiological and/or morphological characteristics set forth in one or more of Tables 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14, as determined at the 5% significance level, when grown in the same location under the same environmental conditions as variety DH-056.149.

4. A Brassica carinata seed produced by crossing Brassica carinata plants and harvesting the resulting Brassica carinata seed, wherein at least one Brassica carinata plant is the plant of claim 2.

5. A method of producing a Brassica carinata variety derived from Brassica carinata variety DH-056.149, the method comprising (c) crossing a plant of Brassica carinata variety DH-056.149 with a different Brassica carinata plant having a desired trait to produce F1 hybrid seed; and (d) growing the resultant F1 hybrid seed and selecting one or more F1 hybrid plants having the desired trait and at least a portion of the genetic make up of Brassica carinata variety DH-056.149.

6. The method of claim 5, further comprising the steps of (a) backcrossing the selected F1 hybrid plants with plants of variety DH-056.149, or with the different Brassica carinata plant having a desired trait, to produce backcross progeny seed; (b) growing the resultant backcross progeny seed and selecting backcross progeny plants that have the desired trait and at least a portion of the genetic make up of Brassica carinata variety DH-056.149; and (c) repeating steps (a) and (b) on the selected backcross progeny plants to a maximum of 10 generations to produce a progeny Brassica carinata plant derived from Brassica carinata variety DH-056.149, wherein the progeny Brassica carinata plant comprises the desired trait and at least a portion of the genetic make up of Brassica carinata variety DH-056.149.

7. The method of claim 5, further comprising the steps of (a) self-pollinating the selected F1 hybrid plants to produce further progeny seed; (b) growing the further progeny seed and selecting further progeny plants that have the desired trait and at least a portion of the genetic make up of Brassica carinata variety DH-056.149; and (c) repeating steps (a) and (b) on the selected further progeny plants to a maximum of 10 generations to produce a progeny Brassica carinata plant derived from Brassica carinata variety DH-056.149, wherein the progeny Brassica carinata plant comprises the desired trait and at least a portion of the genetic makeup of Brassica carinata variety DH-056.149.

8. An F1 hybrid plant grown from the F1 hybrid seed produced by the method of claim 5, wherein the F1 hybrid plant has a desired trait and at least a portion of the genetic make up of Brassica carinata variety DH-056.149.

9. A method for producing a Double Haploid variety comprising: (a) isolating a flower bud of the F1 plant of claim 8; (b) dissecting out a haploid microspore; (c) placing the haploid microspore in culture; (d) inducing the microspore to differentiate into an embryo and subsequently into a plantlet; (e) identifying whether the plantlet contains a diploid chromosome number, wherein the diploid chromosome number occurred through chromosome doubling; and (f) continuing to grow the plantlet if it contains a diploid chromosome number, wherein the Double Haploid variety comprises at least a portion of the genetic makeup of Brassica carinata variety DH-056.149.

10. A cell, seed, plant, or plant part of the Double Haploid variety produced by the method of claim 9.

11. A cell of a Brassica carinata plant or plant part of claim 3.

12. A tissue culture of protoplasts or regenerable cells of the cell of claim 11.

13. The tissue culture of protoplast or regenerable cells of claim 12, wherein the protoplasts or regenerable cells are produced from a tissue selected from the group consisting of leaves, pollen, embryos, roots, root tips, pods, flowers, ovules, and stalks.

14. A Brassica carinata plant regenerated from the tissue culture of claim 12, wherein the plant has the physiological and/or morphological characteristics set forth in one or more of Tables 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14, as determined at the 5% significance level, when grown in the same location under the same environmental conditions as variety DH-056.149.

15. A method of producing a commercial plant product comprising growing the plant of claim 2 to produce a commercial crop and producing the commercial plant product from the commercial crop.

16. A method of producing a commercial plant product comprising growing the F1 hybrid plant of claim 8 to produce a commercial crop and producing the commercial plant product from the commercial crop.

17. A method of producing a commercial plant product comprising growing a plant of the Double Haploid variety of claim 10 to produce a commercial crop and producing the commercial plant product from the commercial crop.

18. A commercial plant product produced from a Brassica carinata plant of variety DH-056.149, wherein the commercial plant product comprises oil, meal, protein isolate or biofumigant.

19. A commercial plant product produced from the Brassica carinata plant of claim 3, wherein the commercial plant product comprises oil, meal, protein isolate or biofumigant.

20. A commercial plant product produced from the F1 hybrid plant of claim 8, wherein the commercial plant product comprises oil, meal, protein isolate or biofumigant.

Description

DETAILED DESCRIPTION

[0273] In one aspect, the present invention relates to cells, seeds, plants, and plant parts of Brassica carinata variety DH-056.149, alternatively carinata variety DH-056.149, variety DH-056.149, or DH-056.149, for which a representative sample of the seed has been deposited under NCIMB Accession number 43000.

[0274] In another aspect, the present invention relates to Brassica carinata plant or plant part produced or derived from seeds, plants, and plant parts of Brassica carinata variety DH-056.149, as well as to all progeny of Brassica carinata variety DH-056.149 produced by one or more breeding, mutagenesis, tissue culture, or genetic modification techniques and having essentially all the physiological and morphological characteristics of a plant or plant part of Brassica carinata variety DH-056.149 when grown in the same location under the same environmental conditions as variety DH-056.149.

[0275] In another aspect, the present invention relates to Brassica carinata plant or plant part produced or derived from seeds, plants, and plant parts of Brassica carinata variety DH-056.149, as well as to all progeny of Brassica carinata variety DH-056.149 produced by one or more breeding, mutagenesis, tissue culture, or genetic modification techniques and having the physiological and/or morphological characteristics set forth in one or more of Tables 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14, as determined at the 5% significance level, when grown in the same location under the same environmental conditions as Brassica carinata variety DH-056.149.

[0276] The present invention also relates to any plants produced or derived from seeds, plants, and plant parts of Brassica carinata variety DH-056.149, that exhibit carinata character.

[0277] For the purpose of the present invention, plants exhibiting carinata character have an erect, upright bearing, are highly branching, with well-developed and aggressive tap root systems (Barro and Martin, 1999). Leaves are generally wide elliptic in shape with weak-medium dentation, medium glaucosity, and very sparse pubescence. Seeds are globose, 1-1.5 mm in diameter and finely reticulated (Mnzava and Schippers, 2004) and vary from yellow to yellow-brown to brown in colour (Getinet 1987; Rahman and Tahir, 2010).

[0278] Plants exhibiting carinata character may be produced from the seed of Brassica carinata variety DH-056.149; a representative sample of the seed has been deposited under NCIMB Accession number 43000 or from any cell, plant or plant part produced from the seed of Brassica carinata variety DH-056.149, using one or more (conventional) plant breeding technologies, cell culture or tissue culture technologies, and/or transgenic technologies.

Plant Breeding Technologies

[0279] Critical to the development and breeding of any crop is the ability to make use of genotypic and phenotypic diversity. Breeding strategies make use of the plant's method of pollination: self-pollination, where the pollen from one flower is transferred to the same or another flower on the same plant or a genetically identical plant; sib-pollination, when individuals with the same family or line are used for pollination; or cross-pollination, where the pollen comes from a flower on a genetically different plant from a different family or line.

[0280] For practical application, a breeder initially selects and crosses two or more parental lines, followed by repeated selfing and selection to produce many unique genetic combinations. For a newly developing crop such as carinata, it is necessary to be able to obtain a sufficient pool of genetic material to identify genetic backgrounds more adapted to target geographies (i.e. a better starting point), as well as variation for traits of interest. This allows for crossing or other modifications to be done to identify genetic combinations superior to the types already tested. Thus, an important initial objective is the collection and characterization of as large of a collection of genetic backgrounds as possible, for each target geography. Parental lines are selected based on breeding priorities and the unique combination of traits available in potential crossing parents. The selection of parents of these crosses is critical to the effectiveness of a breeding program. Parental lines may be closely or distantly related lines of a single plant species or may be two different species of the same genus.

[0281] Crossing, of (inbred) parental lines, by sexual hybridization, is typically done manually in controlled conditions. Often, two or three rounds of crossing are needed to accumulate beneficial alleles into a single genetic background. This includes evaluating offspring of a cross, selecting the most desirable (inbred) lines as future parents, and making the next round of parental selection based on priority targets. Theoretically, billions of different genetic combinations can be produced through a combination of mutagenesis, selfing, and crossing. Since the breeder has no direct control at the cellular level, two breeders will never independently develop the same variety of carinata plants have the same traits.

[0282] In each cycle of selection and evaluation, the breeder selects germplasm to advance to the next generation by growing individual plants in the chosen geography, soil and climate conditions and collecting phenotypic data reflective of actual performance that would be realized by a seed producer. Traits collected focus on those that would be of agronomic or economic benefit in the crop. Examples of traits characterized in a carinata breeding program include, but are not limited to, early plant vigor, plant height, branching habit, days to flower, silique density, flower petal color, pod size, reaction to heat and water stress, disease susceptibility, and pod shatter tolerance.

[0283] In a typical carinata breeding program, the breeder initially selects and crosses two or more parental Brassica carinata lines, followed by repeated selfing and selection to produce many unique genetic combinations, which are evaluated for overall agronomic potential as well as specific traits. Such recurrent selection can be used to improve a population of either self- or cross-pollinating Brassica. Intercrossing of several parents creates a genetically variable population of heterozygous individuals and the best plants are selected based on individual superiority, outstanding progeny and/or excellent combining ability. New populations are created by further intercrossing and selection. This method is useful for the improvement of quantitatively inherited traits controlled by numerous genes.

[0284] Backcrossing may be used to transfer genes for a simply inherited, highly heritable trait from a donor parent to the recurrent parent. After the initial cross, individual plants with the desired trait of the donor parent are selected and backcrossed to the recurrent parent for several generations. The resulting progeny are expected to have the attributes of the recurrent parent and the desired trait from the donor parent. Backcrossing may be used in conjunction with pedigree breeding. Pedigree breeding and recurrent selection methods are used to develop varieties from breeding populations. Pedigree breeding starts with the crossing of two genotypes, each with one or more desirable characteristics that is lacking in the other or that complement each other. Additional genotypes can be included in the breeding population if the original parents do not provide all the desired characteristics. In the pedigree method of breeding, five or more generations of selfing and selection may be used. For example, crossing of the two initial parents (the donor and recurrent parents) produces an initial F1 population, from which an F2 population is produced by selfing one or several F1 plants or by intercrossing two F1 plants (sib mating). Selection of hybrids with desired combination of traits may be conducted with the F2 population, or with the F3 or subsequent population.

[0285] Plants that have been self-pollinated and selected for several generations become homozygous at nearly all gene loci and produce a uniform population of breeding progeny or inbred lines. Subsequent crosses with two different homozygous (inbred) lines produces a uniform population of hybrid plants that may be heterozygous for many gene loci. Crossing two plants each heterozygous at a number of gene loci will produce a population that differ genetically and will not be uniform.

[0286] Doubled haploid (or DH) technology allows for the generation of completely homozygous lines, which are a combination of genes of the parental lines, in a single generation. This accelerates the process of inbred line development dramatically as the process from seeding of parental lines to obtaining seed for a resulting inbred population from those parents will generally take about 18 months. To achieve a highly homozygous line using traditional self-pollination generally will take five or six growth cycles which, in the case of carinata, would represent three years or six growth cycles, with two cycles completed per year. In DH technology, using appropriate in vitro conditions, haploid microspores from an F1 plant can be induced to differentiate into diploid embryos and subsequently plantlets. This technique typically relies on a percentage of regenerated plants to undergo spontaneous doubling (usually in the range of 20 to 60% of plants, depending on several factors), whereas the remaining plants will remain haploid and sterile. To increase the efficiency of space used for seed increase, such as in the greenhouse or field, a flow cytometer is used to distinguish at an early stage the chromosome content n or 2n of each plant. Thus, the sterile plants can be discarded at an early stage.

[0287] In some instances, a desirable trait may not reside within the species of interest, specifically Brassica carinata. In that case, it may be possible to transfer the trait via interspecific or wide crossing. For interspecific crossing, one parent is Brassica carinata variety DH-056.149 and the second parent may be a different species of Brassica, including but not limited to B. juncea (L). Czern. (brown mustard), B. napus L. (rape, Argentine canola), B. nigra (L) W. D. J. Koch (black mustard), B. rapa L. (field mustard, Polish canola), and B. oleracea L. (cabbage, broccoli, cauliflower, brussels sprouts, kohlrabi and kale). In other instances, the other Brassicaceae species including but not limited to Brassica alba, Brassica hirta, Brassica juncea, Brassica napus, Brassica nigra, Brassica oleracea, Brassica rapa, Sinapus alba, and Camelina sativa.

[0288] The methodology for performing interspecific crosses is similar to that described for within-species crosses described above. However, unlike intraspecific crosses, the likelihood that the resulting progeny will produce viable seed is very low and thus represents a formidable challenge to the success of this technique. To overcome this potential block, embryo rescue techniques are often employed to recover viable offspring from the cross. Essentially, this relies on the progeny surviving until the embryogenic stage at which point it can be dissected from the silique and placed into artificial growth medium. Under appropriate conditions, the cultured embryo can survive and be induced to differentiate into a plantlet, which can be grown into a mature plant. Successive rounds of embryo rescue may be needed until inbred progeny, or backcross-derived progeny, are stable and can produce fertile offspring without intervention. Often molecular markers, where available, are used to trace a specific allele from a related species into an adapted background in the target species, using repeated cycles of backcrossing.

[0289] In addition, to minimize the relative proportion of the donor genome from the non-carinata species, several rounds of back-crossing of the rescued plants with the Brassica carinata parent may be required to generate progeny having carinata character and the desired trait, which are stable and can produce fertile offspring.

Selection

[0290] Breeding nurseries are often the first cycle of evaluation of breeding populations. Generally, nurseries utilize single or paired rows with frequent checks (i.e. the best available commercial germplasm for a specified geography.

[0291] Various methods are used to screen breeding populations for individual plants with desired characteristics. Single seed descent procedure refers to planting a segregating population, harvesting a sample of one seed per plant, and using the one-seed sample to plant the next generation. When the population has advanced from the F2 to the desired level of inbreeding, the plants from which lines are derived will each trace to different F2 individuals. Since the number of plants in a population declines each generation due to failure of some seeds to germinate or produce seed, not all the F2 plants will be represented by a progeny in the final generation.

[0292] A multiple-seed procedure is when pods form each plant in a population are harvested and threshed together to form a bulk. Part of the bulk is used to plant the next generation and part is put in reserve. The advantage of this method is that it is faster to harvest seed with a machine than to remove one seed from each plant by hand. If desired, Double(d) Haploid methods can be used to extract homogeneous (homozygous) lines.

Marker Assisted Selection and Marker Assisted Breeding

[0293] A breeding program can make use of marker assisted selection (MAS) and marker assisted breeding (MAB) technologies to accelerate the successful outcome of a breeding project. These techniques enable the identification of lines carrying a trait of interest in the laboratory, while other lines not containing a marker of interest can then be discarded at an early stage. These methods can also increase the efficiency of a program, as the lines being evaluated in the field have a greater probability of meeting seed quality or other criteria. MAS and MAB methods rely on the existence of a dense set of genetic markers for the species of interest. Genetic markers are the unique sequences that may be found in allelic forms of genes, distinguishing one allele from another. Like genes themselves, they can be transmitted to progeny in a Mendelian fashion and can thus be used to follow the movement of specific alleles from parents to progeny.

[0294] Molecular markers can be used in Quantitative Trait Loci (QTL) mapping whereby selection of plants with desired trait(s) is assisted by markers known to be closely linked to alleles that have measurable effects on a quantitative traiti.e., accumulation of markers linked to positive effecting alleles or elimination of markers linked to negative effecting alleles in the plant genome. Markers can also be used to select for the genome of the recurrent parent and against the markers of the donor parent. This can minimize the amount of genome from a donor parent that remains in the selected plants and/or the number of back crosses to the recurrent patent.

[0295] Other types of genetic markers in common use by persons skilled in the art include, but are not limited to, Restriction fragment length polymorphisms (RFLP), Random Amplified Polymorphic DNA (RAPD), Amplified Fragment Length Polymorphism (AFLP), Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Simple Sequence Repeats (SSRs), and Single Nucleotide Polymorphisms (SNPs).

Mutagenesis

[0296] Another method of creating genetic variation and capturing beneficial changes in a heritable fashion is through mutagenesis breeding. This method is often carried out via chemical means or ionizing radiation and is typically focused either on a microspore or on a whole seed level. In Brassica breeding, some common forms of mutagens used have been chemical agents such as ethyl methanesulfonate (EMS) or N-ethyl-N-nitrosourea (ENU), or high levels of ionizing radiation (x-ray or gamma irradiation or exposure to UV light). EMS produces random point mutations via low frequency methylation of guanine residues in genomic DNA. This results in altered Watson-Crick base pairing such that the affected base pairing is converted from G-C to A-T. ENU is also an alkylating agent that preferentially modifies thymine residues converting A-T to G-C. Ionizing radiation may affect DNA in many ways but typically the mutations are double strand breaks leading to deletions and frameshift mutations that are frequently inactivating.

[0297] To carry out this technique, seedlings or microspores are exposed to the mutagenic agent and the surviving fraction are allowed to develop into mature plants. In some cases, the mutagenized plantlets or embryos (in the case of microspore mutagenesis) may be exposed to selection in order to enrich for a particular phenotype. For example, mutagenesis has been used to develop plants that are resistant to the actions of specific herbicides; in this instance the developing plantlets or microspores can be grown in vitro in the presence of the herbicide(s) of interest in order to select for those plants with the appropriate mutations conferring resistance. The advantage of the microspore mutagenesis of the seed approach is that the resultant DH plants can be used to derive pure and homozygous plant lines where all induced mutations, whether dominant or recessive, would be expressed. Mutagenesis has been used to develop Brassica varieties with resistance to various herbicides, altered seed oil profiles and increased tolerance to disease and abiotic stress.

Genetic Transformation and Transgenic Technologies

[0298] In instances where unique and valuable traits are known to be available in distant plant or in non-plant species that cannot be transferred to Brassica carinata via classical breeding, and where the genes for those traits have been cloned, a breeding program may employ genetic transformation techniques, or transgenic technologies, to stably transfer those genes into this species. Transfer of cloned genetic elements into B. carinata have been achieved via a number of means, including PEG-mediated DNA uptake into protoplasts (Johnson, et al., 1989), electroporation into protoplasts (Fromm, et al., 1985), ballistic infiltration using DNA coated microprojectiles (Finer, et al., 1999), Agrobacterium-based vector infiltration (Babic, et al., 1998), and infection using plant virus-based vectors (Gleba, et al., 2004). Aside from having the genes of interest in cloned form, the other requirements include having the genes cloned into a suitable vector to allow for their propagation in an appropriate bacterial system, as well as their packaging in appropriate viral and Agrobacterium strains if transformation utilizes an infectious route of transfer. Once transferred, the gene(s) of interest would also require appropriate plant-based promoters, enhancers and terminators to allow for the correct temporal and tissue specific pattern of expression for the heterologous gene. Finally, to select for those rare events where the heterologous gene expression unit has been successfully transferred into the plant genome, a selectable marker may be introduced, either physically linked to the heterologous gene of interest or co-transformed with the gene of interest at a suitable ratio to favor co-insertion.

[0299] The selectable marker may consist of a gene that can confer resistance to a particular herbicide or antibiotic that would otherwise kill the plant, a gene that may confer a growth advantage, a gene that may alter a response to plant hormones, or a gene that expresses a fluorescent protein that can allow transformed cells to be easily visualized. Examples of selectable markers conferring resistance to antibiotics that have been successfully used in Brassica transformation are the NPTII gene (Bevan 1984; Datla, et al., 1992), encoding an enzyme conferring resistance to the antibiotic kanamycin, and the HPT gene encoding an enzyme conferring resistance to the antibiotic Hygromycin (Rothstein, et al., 1987). Examples of selection markers based on conferring tolerance to herbicides and successfully used in Brassica transformation are the BAR (Thompson, et al., 1987) and PAT (Wohlleben, et al., 1988) gene products, which encode phosphinothicine acetyltransferase and confer resistance to glufosinate (bialaphos) or L-PPT, and the AHAS gene product encoding acetohydroxyacid synthase enzyme conferring resistance to imidazolinones (Miki, et al., 1990). Other plant selectable markers have been developed whose actions are not based on conferring resistance to toxic compounds per se but instead allow survival in the presence of nutrients not normally metabolized by the wildtype organism.

[0300] Transformation cannot only be used to introduce heterologous genes into the genome of carinata plants, it can also be used to introduce nucleic acid constructs that are designed to modulate the expression of endogenous genes. Nucleic acid constructs encoding antisense RNA or RNAi sequences (Tang and Galili, 2004) can be used to interfere or knock down the expression of endogenous genes to extremely low levels, simulating the effect of a null mutation at the endogenous locus. This of course relies on the continuous stable expression of the antisense RNA or RNAi to be effective. In amphidiploid Brassica species such as napus, juncea and carinata, multiple copies of genes from the contributing ancestral species may create a high level of functional redundancy such that a single mutation in one of the homologues may not be sufficient to confer a phenotype. However, by using an RNAi or antisense approach, where the interfering RNA is derived from conserved sequences, one may conceivably be capable of targeting all the expressed homologues and achieving a functional knockdown effect.

[0301] Examples of modifications that can be introduced into Brassica carinata using genetic transformation include, but are not limited to, [0302] genes that control pollination, hybrid seed production, or male-sterility; [0303] genes encoding resistance to pathogens and insect pests (plant disease resistance gene(s); gene(s) conferring resistance to fungal pathogens; natural or synthetic Bacillus thuringiensis (Bt) protein or a derivative thereof; an insect-specific hormone or pheromone coach or a mimetic based thereon, or an antagonist or agonist thereof; an insect-specific peptide that disrupts the physiology of the affected pest; an enzyme responsible for hyperaccumulation of a non-protein molecule with insecticidal activity; a membrane permease, a channel former or a channel blocker; a viral-invasive protein or a complex toxin derived therefrom; an insect-specific antibody or an immunotoxin derived therefrom; genes involved in the Systemic Acquired Resistance (SAR) Response and/or the pathogenesis related genes; antifungal genes; detoxification genes, such as for fumonisin, beauvericin, moniliformin, and zearalenone, and their structurally related derivatives; cystatin and cysteine protease inhibitors; Defensin genes; genes that confer resistance to Phytophora Root Rot, such as the Brassica equivalents of the Rps 1, Rps 1-a, Rps 1-b, Rps 1-c, Rps 1-d, Rps 1-e, Rps 1-k, Rps 2, Rps 3-a, Rps 3-b, Rps 3-c, Rps 4, Rps 5, Rps 6, Rps 7, and other Rps genes); [0304] genes that confer resistance to a herbicide (mutant ALS and AHAS enzymes to confer resistance to imidazolinone or sulfonylurea herbicides; mutant 5-enolpyruvylshikimate-3-phosphate synthase (EPSP) and aroA genes conferring resistance to glyphosate; phosphinothricin acetyl transferase (PAT) and Streptomyces hydroscopicus phosphinothricin-acetyl transferase (BAR) conferring resistance to other phosphono compounds such as glufosinate; ACCase inhibitor-encoding genes conferring resistance to pyridinoxy or phenoxy propionic acids and cyclohexones; pabA and gs+ genes conferring resistance to triazine; nitrilase gene conferring resistance to benzonitrile; acetohydroxy acid synthase, which has been found to make plants resistant to multiple types of herbicide); [0305] genes that confer or contribute to altered fatty acids (down-regulation of stearoyl-ACP desaturase to increase stearic acid; elevating oleic acid via FAD-2 gene modification and/or decreasing linolenic acid via FAD-2 gene modification; altering conjugated linolenic or linoleic acid content; altered LEC1, AGP, Dek1, Superal1, mi1 ps, various lpa genes such as lpa 1, lpa3, hpt, or hggt); [0306] genes that confer or contribute to altered phosphorus content (introduction of a phytase-encoding gene; up-regulation of a gene that reduces phytate content); [0307] genes that confer or contribute to altered carbohydrates, antioxidant, or essential seed amino acids; [0308] genes that create a site for site specific DNA integration, such as the introduction of FRT sites that may be used in the FLP/PRT system and/or Lox sites that may be used in the Cre/Loxp system; [0309] genes that affect abiotic stress resistances (including, but not limited to flowering, pod and seed development, enhancement of nitrogen utilization efficiency, altered nitrogen responsiveness, drought resistance or tolerance, cold resistance or tolerance, and salt resistance or tolerance, and increase yield under stress}.

Gene Editing

[0310] More recently, several novel approaches have been developed which offer the ability to manipulate the plant genome in a targeted way. Collectively known as gene editing technology (Petolino, et al., 2010; Woo, et al., 2015, Sauer, et al., 2016), the technologies share several important similarities: [0311] they offer the ability to introduce small or large gene deletions or insertions in the plant genome by means of targeted double strand breaks at precise genomic locations; [0312] they allow for the insertion of small inactivating mutations into specific genes of interest; [0313] they permit the replacement of an entire gene or gene segment by a modified counterpart; [0314] they allow for the insertion of a heterologous gene in a specific genomic location which may represent a preferred site for regulated gene expression. (i.e. downstream of a known endogenous promoter/enhancer).

Locus Conversion

[0315] The Brassica carinata variety described herein represents a new base genetic line into which a new locus or trait my be introduced using transgenic technologies or back-crossing. The term locus conversion refers to the product of such an introgression. For example, a donor parent having a specific desirable trait may be crossed with an inbred variety, the recurrent parent, which has overall good agronomic characteristics yet lacks the desirable trait. The progeny of this cross is then mated back to the recurrent parent followed by selection of the desired trait from the donor parent.

[0316] A locus converted plant cell of a locus converted plant is obtained by introducing one or more locus conversion in the Brassica carinata variety described herein. The locus converted plant cell is therefore identical to a cell from the Brassica carinata variety described herein except for the one or more locus conversion and the locus converted plant exhibits essentially all the physiological and morphological characteristics of the Brassica carinata variety described herein. The locus conversion can confer a trait selected from the group consisting of male sterility, disease resistance, fungal resistance, pest resistance, herbicide tolerance, abiotic stress tolerance, and altered metabolism. Both naturally occurring and transgenic DNA sequences may be introduced through backcrossing. Molecular marker assisted breeding or selection maybe use used to reduce the number of backcrosses need to achieve the backcross conversion.

[0317] A locus conversion of the Brassica carinata variety described herein will otherwise retain its genetic integrity. For example, a locus converted plant of the Brassica carinata variety described herein will comprise at least 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of its base genetics of the starting variety. A locus converted plant of the Brassica carinata variety described herein will therefore have at least one, and possibly two or three physiological or morphological characteristics which are different from those of a locus converted plant of the Brassica carinata variety described herein and otherwise has all the physiological or morphological characteristics of a locus converted plant of the Brassica carinata variety described herein.

Male Sterility

[0318] To produce and test hybrid combinations, some form of pollination control system must be employed. The most widely used pollination control system to achieve this in Brassica species is the method of Ogura (Ogura, 1968) adapted and modified by scientists at INRA, which is based on control of pollination by use of male cytoplasmic sterility. In cytoplasmic male sterile (CMS) plants, a mitochondrial gene mutation interferes with the flower's ability to produce viable pollen but does not affect the functionality of the flower's female components. Because the mutation is within the mitochondrial genome, it is transmitted maternally through the cytoplasm. CMS plant lines, termed A lines, can only produce viable seed via outcrossing with non-CMS plants; however, the F1 plants derived from the seed of such a crossing will also exhibit the CMS phenotype and will still not produce viable seed by self pollination.

[0319] The Ogura CMS trait, originally identified in Japanese radish varieties (Ogura, 1968), was transferred to Brassica oleracea via backcrossing (Bannerot, et al., 1977) then subsequently transferred to Brassica napus. These initial Brassica napus A lines were found to be deficient in terms of their sensitivity to cold and their tendency to exhibit leaf chlorosis, as well as fertility issues, which were ultimately found to be related to interactions with chloroplasts derived from the radish cytoplasm (Pelletier et al., 1983). Using a cell fusion approach, improved Brassica napus A lines were subsequently developed that overcame these Initial deficiencies (Pelletier, et al., 1987). A-lines can be maintained by crossing them with B lines (lines that are genetically identical to A lines except that their cytoplasm is not CMS).

[0320] For production of fully fertile hybrid seed, CMS lines must be crossed with lines that contain a restorer function gene (Rf) which can restore the ability of CMS plants to produce viable pollen. Plant lines with the RF function stably incorporated and homozygous (termed RF lines) can be crossed with CMS lines and the F1 seed harvested from the CMS plants will represent a true hybrid and which will have the ability to produce fully fertile offspring capable of self pollination, a prerequisite for commercial grain production.

[0321] Such an Rf activity, first identified in radish, was introduced into Brassica napus (Heyn, 1976). It was subsequently shown that these lines were affected by fertility problems (Heyn, 1978; Pellan-Delourme and Renard, 1988). Intensive backcrossing of these initial lines eventually yielded material with improved fertility such that they could be used practically as Rf lines (Delourme, et al., 1991). It was further noted, however, that hybrid seed produced from crosses with the early versions of Brassica napus Rf lines often had high glucosinolate levels (Delourme, Foisset et al., 1998) that made generation of canola quality hybrids problematic. Efforts to develop improved Rf lines by reducing the amount of extraneous radish genetic material has resulted in newer generations of Brassica napus (Primard-Brisset, et al., 2005) and Brassica juncea (Tian, et al., 2014) Rf line families of sufficiently quality to be used in commercial hybrid seed production. Seed of Rf lines can be maintained by allowing self crossing of plants grown in isolated tents or fields.

[0322] A, B and Rf lines are thus the prerequisite components of an expandable production system for F1 hybrid seed. In practice, for production of pure hybrid seed: [0323] a. the hybrid production fields are required to be reproductively isolated from extraneous sources of Brassica pollen; [0324] b. RF lines are seeded at a predetermined optimal ratio relative to A lines and may be kept spatially separated from the A lines so that the RF plants can be readily removed post flowering, and prior to harvest of the A line material; [0325] c. pollinators are absolutely required to allow for maximal pollination of the A lines by CMS pollen donors; [0326] d. for maintenance and expansion of RF stocks, RF seed is planted in isolation (either isolated fields separated from other Brassica crops by great distance, or plots that are reproductively isolated by virtue of a physical barrier such as a tent) and allowed to self pollinate; [0327] e. for maintenance and expansion of A line stocks, A lines and B lines are planted in separate rows in isolated fields or tents (as described above) and pollinators are used to promote pollination of A lines with B line pollen. B lines are cut down after flowering and before seed set and the A line plants are allowed to come to maturity then harvested.
In some cases, a less labor- and cost-intensive alternative to that described in (b) is employed whereby a low ratio of RF plants may be interspersed with the A line plants in the field and harvested together. While there will be some level of adulteration of the harvested hybrid grain with that of the self pollinated Rf plants; however, in practice this level of contamination will not greatly affect the commercial quality of the hybrid seed

[0328] More recently, both the CMS and Rf traits have been transferred to Brassica carinata. The development of a set of genetically diverse CMS and Rf lines in Brassica carinata allows the testing of many combinations for the first time in this Brassica species.

[0329] In a hypothetical test scheme based on the Ogura CMS system, the following steps are carried out: [0330] panels of genetically diverse Brassica carinata varieties are selected from a previously characterized collection of elite germplasm and used for introgression of CMS or Rf traits to produce A and Rf test lines. Diversity can be estimated by comparing differences in molecular marker profiles between candidate lines; [0331] crosses are carried out between selected diverse A and Rf test lines, under conditions described in previous sections; [0332] F1 carinata hybrid seed from test crosses are planted in replicated small plot trials, F1 plants and harvested seed are assessed for a number of traits including, but not limited to, early vigour, yield potential, grain and oil yield per acre, as well as seed quality traits; [0333] patterns of heterosis between pairs A and Rf parents are identified and F1 hybrid seed from promising combinations are studied more extensively in larger, geographically diverse trials; and [0334] hybrid seed from specific combinations may be selected for potential commercial release as a new variety.

Phenotype/Genotype

[0335] To be useful and reliable, a Brassica carinata variety or hybrid must be homogenous and reproducible. While there are a number of analytical methods known to the skilled person for assessing the phenotypical stability of a Brassica carinata variety or hybrid, the traditional method is the observation of phenotypic traits over the life of the carinata plant using data collected from field experiments conducted under the selected geographic, climatic, and soil conditions during one or more growing seasons. Phenotypic characteristics observed include, but are not limited to, traits associated with seed yield (pod density, number of seeds per pod, pod length), oil yield, seed oil quality (GSL and erucic acid content; fatty acid composition), seed protein content, seed protein quality, glucosinolate composition of meal, growth habit, lodging resistance, plant height, and pod shatter resistance. Other phenotypic characteristics that may be observed include, but are not limited to, traits associated with pest tolerance or resistance, cold or frost tolerance, disease tolerance or resistance, herbicide tolerance or resistance, early or late flowering, and/or early or late maturity.

[0336] In some embodiments, Brassica carinata varieties or hybrids useful for commercial crop production or production of commercial products may exhibit traits associated with seed yield, oil yield, seed oil quality, erucic acid content of seed or oil, glucosinolate content of seed, seed protein content, fatty acid composition of oil, glucosinolate composition of meal, meal protein content, growth habit, lodging resistance, plant height, and pod shatter resistance

[0337] In some embodiments, a Brassica carinata variety or hybrid may exhibit multiple traits or phenotypic characteristics that provide agronomic advantages in particular geographies, climates, cropping regimes, and/or soil types. For example, [0338] Brassica carinata varieties for planting in regions with a climate classified as being of tropical moist characteristics, with planting occurring in fall or winter for harvest in spring or summer, may be selected for one or more traits including, but not limited to, superior yield of oil per area planted, shorter time to maturity, improved frost tolerance, improved disease resistance, and resistance to pod shatter. [0339] Brassica carinata varieties for planting in regions with a climate classified as being of cool temperate, dry classification, with planting occurring in spring and harvest in summer or fall, may be selected for one or more traits including, but not limited to, superior yield of oil per area planted, shorter time to maturity, tolerance to drought, improved disease resistance, and resistance to pod shatter. [0340] Brassica carinata varieties for planting in regions with a climate classified as being of warm temperate, moist characteristic for planting in fall or winter for harvest in spring or summer, may be selected for one or more traits including, but not limited to, superior yield of oil per area planted, shorter time to maturity, tolerance to drought, improved disease resistance, and resistance to pod shatter.

[0341] Other traits for which Brassica carinata varieties may be selected include, but are not limited to, Alternaria resistance, days to flowering, depth of canopy, duration of flowering, end of flowering, flower petal coloration, frost tolerance, herbicide resistance, leaf colour, leaf glaucosity, leaf length, leaf number of lobes, leaf width, maturity, seed maturity, number of seed-bearing pods, pod (silique) beak length, pedicel length, petiole length, plant height, plant length, pod (silique) length, pod (silique) width, primary raceme length, recovery from frost damage, Sclerotinia resistance, seed colour, seeds per pod, seed weight (thousand seed weight or TKW), and stem color.

[0342] Genotype assessment(s) can be used to confirm the homogeneity and reproducibility of a Brassica carinata hybrid, to identify plants of the same variety or related variety, and to confirm the pedigree of the plant. Techniques known to those skilled in the art for the analysis and comparison of plant genotype include, but are not limited to, whole genome sequencing, Restriction fragment length polymorphisms (RFLP), Random Amplified Polymorphic DNA (RAPD), Amplified Fragment Length Polymorphism (AFLP), Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Simple Sequence Repeats (SSRs), and Single Nucleotide Polymorphisms (SNPs).

[0343] The Brassica carinata variety of the present invention has shown uniformity and stability for all traits described in the variety description information in Table 1, which include morphological, agronomic, and quality traits for Brassica carinata variety DH-056.149. The detailed phenotypic information provided in Table 1 is based on data collected in field experiments using conventional agronomic practices. For comparative purposed, Brassica carinata varieties AAC-A120 and AGRO44-312D-HP11 (WO2017/181276A1; henceforth referred to as AG044-HP11 or HP11) were similarly grown as comparative check varieties in replicated experiments, and observations were recorded for the various morphological traits for Brassica carinata variety DH-056.149 and the comparative check varieties.

Disease

[0344] Varieties of Brassica carinata may be susceptible to a number of pathogens that commonly infect Brassica species including, but not limited to, the bacteria and fungi listed below:

TABLE-US-00001 Disease Organism Disease Organism Bacterial leaf spot Pseudomonas Light leaf spot Pyrenopeziza syringae brassicae Bacterial leaf rot Erwinia marginalis Powdery mildew Erysiphe polygoni Bacterial soft rot Pseudomonas Root rot complex Rhizoctonia solani, marginalis Fusarium spp., Pythium spp. Black rot Xanthomonas Seeding disease Rhizoctonia solani, campestris complex Fusarium spp., Pythium spp. Alternaria black spot Alternaria spp Sclerotonia white Sclerotonia stem rot sclerotiorum Alternaria grey leaf Alternaria brassicae Aster yellow Phytoplasma spp. spot Alternaria brassicola Alternaria pos spot Alternaria brassicae Damping off Phytophthora Alternaria brassicola cactorum Pythium spp. Black leg Leptosphaeria White leaf spot Mycosphaerella maculans capsellae Leptosphaeria biglobosa Black root Aphanomyces Grey stem Mycosphaerella raphanin capsellae Clubroot Plasmodiophora Wirestem and Rhizoctonia solani brassicae girdling root rot Downey mildew Peronospora White rust Albugo candida parasitica Fusarium wilt Fusarium oxysporum Verticillium wilt Verticillium Fusarium avenaceum longisporum

[0345] Examples of economically significant fungal diseases of Brassica species and mustard oilseeds include [0346] a. Sclerotinia stem rot is caused by a fungus whose spores infect Brassica species primarily during flowering stages and whose incidence is associated with periods of high humidity. Lesions are formed on the stems which can eventually kill the plant. Fungicides are available which can control the severity of the infection but must be applied at specific periods of the plant lifecycle (i.e. at early to mid-flowering) for best effect. Often multiple applications within this window of time are necessary. [0347] b. Alternaria is a fungal disease of Brassica species that affects plants at all growth stages from early seedling through to maturity although mature plants are more susceptible. The greatest economic impact is on grain yield and quality. Foliar fungicide application during the late flowering stage is an effective way to mitigate the more detrimental effects of the disease on grain yield and quality. [0348] c. Blackleg is caused by a fungal pathogen Leptosphaeria maculans of Brassica oilseed crops, which infects plants at all stages, but early stage infections have the most serious consequences, often culminating in plants with necrotic lesions on their lower stems that can virtually sever the plants at the base. Fungicides are only partially effective, having a minor protective effect when applied at an early plant growth stage. [0349] d. Clubroot is caused by a soil borne fungus-like pathogen (Plasmodiophora brassicae) that affects the roots of Brassica oilseed crops. The spores can persist for long periods in the soil and there is currently no effective fungicidal treatment. Management may require using rotations which limit the frequency of Brassica planting.

[0350] The development of disease tolerant or disease resistant carinata varieties is important for achieving high yields of seed, oil, and other products from Brassica carinata. Conventional methods for control of microbial disease may employ one or more of chemical control, disease resistance, and culture control procedures such as crop rotation, liming, and use of bait crops.

[0351] When producing a commercial crop or a group of cultivated Brassica carinata plants in a field, harmful microorganisms can be controlled by the application of a composition comprising one or more microbiocidal ingredients, such as a fungicide. Fungicides comprise a diverse set of chemical agents which can prevent or reduce the severity of plant infection by pathogenic fungi. There are numerous classes of fungicides. FRAC (Fungicide Resistance Action Committee; http://www.frac.info/home) lists 12 classes based on the different biochemical pathways that the fungicides within a class targets, as well as a 13.sup.th class which comprises fungicides with unknown modes of action. Fungicides are also distinguished by their modes of delivery and sites of action: some fungicides are sprayed onto plant surfaces, some are applied to the soil surfaces either in granular form or as a liquid flooding the soil surface, while others are applied as seed treatments. Fungicides applied to the seeds or soils tend to be absorbed via the roots and are transported to all plant tissues via xylem. Fungicides that are foliar can be either locali.e., protecting only the surfaces that they contact, systemici.e., absorbed by the upper plant surfaces but then transported by xylem to all above ground tissues, or partially systemici.e., they can be locally absorbed but can only be transported short distances to protect a somewhat more extensive surface than the initial point of fungicide contact. Fungicides can help mitigate the risk of losses incurred by fungal infection, but the costs of fungicide spraying are significant enough to require cost benefit and risk assessment type analyses to be carried out before deciding to proceed.

[0352] When producing a commercial crop or a group of cultivated Brassica carinata plants in a field, harmful microorganisms can be controlled by the application of a composition comprising one or more microbicide or fungicide ingredient. Examples of microbiocides and fungicides useful for disease control for Brassica carinata include, but are not limited to, azoxystrobin, boscalid, fluxapyroxad, pyraclostrobin, picoxystrobin, propiconazole, metconazole, iprodione, prothiaconazole, vinclozolin, carbathiin, thiram, difenoconazole, metalaxyl, sedaxane, fludioxonil, penflufen, trifloxystrobin, and sedaxane.

Insect Pests

[0353] A variety of insect pests may infest and/or cause damage to a developing Brassica carinata plant. Common insect pests of Brassica at the seedling stage include, but are not limited to, aphids (Lipaphis erysimi, Myzus persicae, Brevicoryne brassicae), flea beetles (Phyllotreta cruciferae, Phyllotreta striolata, Psylliodes punctulata), cutworm (Agrotis orthogonia, Euxoa ochrogaster, Feltia jaculifera, Lacimpolia renigrea), and cabbage root maggot (Delia radicum). Common insect pests of Brassica at the flowering and podding stages include, but are not limited to, diamondback moth (Plutella xylostell), Berta armyworm (Mamestra configurata), and cabbage seed pod weevil (Ceutorhynchus obstrictus).

[0354] When producing a commercial crop or a group of cultivated Brassica carinata plants in a field, insect pests may be controlled by the application of a composition comprising one or more insecticide. Insecticides are a group of pesticide compounds designed to reduce or eliminate crop loss due the predation of crop species by insects. Like herbicides and fungicides, insecticides are classified according to their mode of action and the biochemical pathways that they target. One classification scheme (IRAC MoA) advocated by the Insecticide Resistance Action Committee (IRAC; http://www.irac-online.org) lists 29 classes of insecticides grouped by the common biochemical processes and pathways that the insecticide compounds target. Like herbicides and fungicides, insecticide function and persistence can also be influenced by their sites of action, i.e. whether they are only active on the surface of plants as applied, or whether they function as systemic agents. Further differentiation among some insecticide groups may be apparent based on whether they exhibit selectivity for specific insect types due to distinctive aspects of that insect's biology. Given that some insects serve a beneficial role, such as controlling plant pests, serving as plant pollinators and improving the nutrient content of soil, it is important that insecticides not be applied indiscriminately, but rather are used in a way that limits their actions as much as possible to the desired target species. Thus, modalities such as timing of application, amount and route of application, and restrictions on the types of insecticides used and the crops they may be used are all incorporated into the registered usage criteria of insecticide as a means of ensuring their safety and efficacy.

[0355] Insecticides useful for the control of insect pests of Brassica carinata include, but are not limited to, zeta-cypermethrim, zeta-cypermethrim-S-cyanol, lambdacyhalothrin, methoxyfenozide, cyantraniliprole, imidacloprid, thiamethoxam, and clothianidin. Such insecticide can be applied as a seed treatment or as a foliar treatment.

Plant Pests

[0356] Brassica carinata is an aggressive crop and will out-compete many weeds if it establishes well. Some weed species, however, if allowed to establish early and persist, can affect quality and yield of all crops, including carinata. Examples of weeds that can adversely affect yield and quality include cochia, wild mustard, and wild radish. Weed management is thus an important aspect of modern agricultural practice and comprises several different but complementary approaches including physical methods to remove weeds before seed can be set, such as cultivation, tilling and rogueing of fields as well as use of chemical agents or herbicides to suppress or kill weedy species before they become established and/or are able to set and release their seed.

[0357] When producing a commercial crop or a group of cultivated Brassica carinata plants in a field, plant pests can be controlled by the application of a composition comprising one or more herbicide. Herbicides comprise a large group of chemical compounds that interfere with specific biological processes of the plants in such a way as to block their growth and survival. Herbicides are grouped into classes defined by the biological process with which they interact. These can include inhibition of lipid biosynthesis, inhibition of amino acid biosynthesis, hormonal regulation of plant growth, inhibition of photosynthesis, inhibition of nitrogen metabolism, inhibition of plant pigments biosynthesis or function, agents which can disrupt cell membranes and agents which inhibit seedling growth (Sherwani, et al., 2015). In general, different compounds and herbicide classes may display preferential efficacy against certain weedy species. Moreover, some crop species may display more tolerance to certain classes of herbicide than others. Thus, in a particular geographical region, the use of a particular herbicide for weed control may be dictated by the nature of the crop being cultivated and the native weeds encountered in the region. The registered usage also specifies specific methods of application of the herbicide, including recommended concentration of herbicide, use of appropriate diluents, adjuvants, or surfactants, method of delivery (i.e. spray versus granular), timing of application at appropriate crop stage to ensure least crop damage, timing of application and number of applications to ensure optimal weed control, location of application (foliar or soil application), recommended weather conditions for optimal weed control. Some examples of herbicides recommended for use with Brassica carinata grown in SE USA are listed (Seepaul, et al., 2015).

Seed Cleaning

[0358] Cleaning (of) seed or seed cleaning refers to the removal of foreign material from the surface of the seed. Foreign material to be removed includes, but is not limited to, fungi, bacteria, insect material (including insect eggs, larvae, and parts thereof), and any other pests that exist on the surface of the seed. Cleaning (of) seed or seed cleaning also refers to removal of any debris or low quality, infested, or infected seeds, or seeds of different species that are foreign to the sample.

Seed Treatment

[0359] Prior to planting, Brassica carinata a composition may be applied to the seed as a seed treatment. The composition may be applied at any time from harvesting of the seed to sowing of the seed using methods including, but not limited to, mixing in a container, mechanical application, tumbling, spraying, misting, and immersion. The composition may be applied as a liquid, a slurry, a mist, a soak, or a powder. The composition may comprise one or more of a pesticide, fungicide, insecticide, antimicrobial, a bacterial or fungal inoculant for nutrient utilization, plant growth regulator, or plant signalling compound. A general discussion of techniques for application of fungicides to seeds may be found, for example, in chapter 9 of (Jeffs 1978)

Commercial Crops and Commercial Plant Products

[0360] Commercial crop production comprises the steps of seeding, cultivation and harvesting of grain: [0361] Seeding: Brassica carinata can be planted into conventionally tilled soil where conventional tillage or full-tillage comprises a substantial soil inversion repeated several times yearly such that few plant residues remain at the soil surface at the time of seeding. Alternatively, carinata can planted into soil that is maintained under conservation tillage practices whereby the extent and frequency of tillage is substantially reduced with respect to conventional tillage (so-called medium or low tillage soil management) or preferably, it may be no-till planted in standing stubble. Seeding is carried out at a rate designed to achieve plant densities in a range from 80 to 180 plants per square meter. B. carinata should be seeded at a consistent 1.25 to 2.5 cm depth. Brassica carinata is a mid- to long-season crop that requires a slightly longer growing season than other mustard types. Hence seeding early provides the best results. The ideal seeding date depends greatly on geography and weather. In general, soils should be at least 4 C. (40 F.) or higher before planting. In the Canadian Prairies and US northern tier, typical planting occurs in spring between early April to late May. In South Eastern US, typical planting occurs in fall between October and December. In South America, the optimal planting time occurs in fall or winter (i.e. typically between beginning of May to end of June). [0362] Cultivation: For good stand establishment, Brassica carinata requires adequate soil moisture at seeding and through emergence but can tolerate reduced moisture thereafter and stands up well to the semi-arid mid-summer conditions. Brassica carinata is a temperate climate crop but which has been adapted to the more extreme conditions experienced in the southern Canadian prairies and Northern Tier US states. During initial stand formation, carinata can recover from short term frost conditions and tolerates higher heat during flowering and seed set better than other Brassica oilseeds. The fertility requirements of Brassica carinata are similar to other mustards and canola. Adequate availability of the primary macronutrients nitrogen, phosphorous, potassium and sulfur are required to achieve the true yield potential. Lesser amounts of secondary macronutrients, including calcium (Ca), magnesium (Mg) and sulfur (S) and trace amounts of micronutrients (such as boron, copper, Iron, manganese, zinc) may also contribute to optimal plant growth and yield. Fertilizer rates vary with growing zone and soil fertility. [0363] Harvesting is the act of collecting the portion of a plant that has matured sufficiently over the course of a growing season and that has value as a source of food, feed, fibre, feedstock, structural material or as a propagule for the plant itself. Brassica carinata is harvested, for example, by mechanical harvesting, ideally when seed maturity is reached (seed, pods and stalks turn from green to yellow, seed moisture is 9.5 5 or less). Brassica carinata can be combine harvested by straight cutting or, if need be, can be swathed at an early stage, allowed to dry naturally or with the aid of a desiccant, then the dried swath can be combined. Swathing mean cutting near the base of the plant and allowing the plant to lie flat in field for several days to allow the grain to reach the appropriate dryness. However, since Brassica carinata has a sturdy stalk, the preferred method for harvest of carinata is direct combining at maturity, rather than swathing or pushing followed by combining. [0364] Combining: refers to the process of reaping and collecting the seed pods from the matured crop, threshing the seed pods to release the seed (grain), and winnowing to separate and recover the grain from the now empty seed pods, stems, and branches (collectively referred to as chaff). These once distinct operations are today often combined by use of a multifunctional mechanized apparatus, appropriately known as a combine harvester. [0365] Grain, in reference to Brassica carinata, refers to the seed harvested at maturity and sold as a source of oil and meal products.

[0366] Commercial products produced from Brassica carinata seed include, but are not limited to, crushed, non-viable seed, oil, meal, and protein isolate. Production of oil, meal and protein isolate involves multiple steps. Typically, the seeds are cleaned then crushed in a roller mill to generate flakes. The flaked seed then undergoes a cooking process in which it is conveyed to a heated drum where the flakes are cooked at elevated temperatures (typically from 70-90 C.). The cooking helps to reduce the viscosity of the oil to allow for more efficient extraction in subsequent steps. Cooked seed flakes are then pressed in a series of screw presses or expellers which can remove 50-60% of the oil. Aside from the oil, which is removed for further processing, the pressing produces a meal cake that is ideal for solvent extraction. Using several cycles of extraction, the meal cake is treated with a solvent such as hexane to remove the residual oil from the meal. The meal is then transferred to a desolventizer-toaster where it is heated to remove remaining solvent; the final step of the process, called toasting, involves injection of stream into the meal to remove the last traces of solvent. The meal is then cooled and dried by blowing forced air through it. In some cases, the seed can also be processed using a cold press methodology which is similar to above except it does not involve the use of solvent to remove residual oil from the oil cake, resulting in a meal with much higher oil composition. In other cases, meal containing high levels of glucosinolates can be combined with other ingredients and formulated into pellets for use as a biofumigant (U.S. Publication No. 2008/0199451).

[0367] Brassica carinata plants can also be used commercially for biofumigation to reduce the population of disease-causing organisms in the soil. Typically, a field is seeded with Brassica plants early in the planting season, the plants are grown for a time, and the biomass is collected between flowering and before seed set. The plant biomass is then chopped to release the myrosinase enzyme and convert the glucosinolates in the plant biomass to isothiocyanates. The chopped biomass is tilled into the soil prior to seeding of a second crop

Uses of Brassica carinata Variety DH-056.149

[0368] Brassica carinata variety DH-056.149 can be used in accordance with any of the breeding methods described herein, as well as in breeding methods known to those skilled in the art, to produce carinata hybrids or other progeny plants having the desired traits and characteristics of variety DH-056.149.

[0369] The invention is directed to methods for producing carinata seeds, plants and plant parts from a carinata plant produced by crossing a first parent plant with a second parent plant, wherein the first parent plant is Brassica carinata variety DH-056.149 and the second parent plant is also Brassica carinata variety DH-056.149, another Brassica carinata variety, or a variety of another Brassicaceae species. In some embodiments, Brassica carinata variety DH-056.149 may be the male or female parent. In other embodiments, either the first or second parent plant may be male sterile. In some embodiments, the second parent plant comprises a desired trait. In some embodiments, the desired trait is selected from the group consisting of male sterility, disease resistance, fungal resistance, pest resistance, herbicide tolerance, abiotic stress tolerance, and altered metabolism. In other embodiments, the desired trait is herbicide tolerance and the herbicide is selected from, but not limited to, the group consisting of glyphosate, glufosinate, imidazolinones, and auxin analogues such as 2,4-D and dicamba.

[0370] In another aspect, the invention is directed to a method of producing a first generation (F1) hybrid Brassica carinata seed, as well a first generation (F1) hybrid plant grown from such seed, comprising crossing Brassica carinata variety DH-056.149 with a different Brassica carinata plant and harvesting the resultant F1 hybrid Brassica carinata seed, and wherein Brassica carinata variety DH-056.149 is either a female parent or a male parent.

[0371] In another aspect, the invention is further directed to a method for producing a Doubled Haploid (DH) variety comprising isolating a flower bud of the F1 hybrid plant, dissecting out a haploid microspore, placing the haploid microspore in culture, inducing the microspore to differentiate into an embryo and subsequently into a plantlet, identifying whether the plantlet contains a diploid chromosome number, wherein the diploid chromosome number occurred through chromosome doubling, and continuing to grow the plantlet if it contains a diploid chromosome number. In one embodiment, the diploid chromosome number is obtained by chemical or physical means. In another aspect the invention is directed to a plant, plant part, or seed of such Double Haploid variety produced from the F1 hybrid plant resulting from crossing Brassica carinata variety DH-056.149 with a different Brassica carinata plant.

[0372] In another aspect, the invention is directed to producing a Brassica carinata plant, as well as a plant, plant part, cell, or seed therefrom, by crossing Brassica carinata variety DH-056.149 with a second Brassica carinata variety having a desired trait, growing the resultant F1 hybrid seed and selecting one or more progeny plants that have the desired trait, backcrossing the selected progeny plants that have the desired trait with plants of variety DH-056.149 to produce backcross progeny seed, and growing the resultant backcross progeny seed and selecting backcross progeny plants that have the desired trait and at least a portion of the genetic make up of Brassica carinata variety DH-056.149.n In some embodiments, the steps of backcrossing and growing the resulting progeny seed may be repeated 1 to 2 times, 1 to 3 times, 1 to 4 times, or 1 to 5 times, or until the Brassica carinata variety produced from variety DH-056.149 has the desired trait and the physiological and/or morphological characteristics set forth in one or more of Tables 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14, as determined at the 5% significance level, when grown in the same location under the same environmental conditions as variety DH-056.149. In some embodiments, the desired trait is selected from the group consisting of male sterility, disease resistance, fungal resistance, pest resistance, herbicide tolerance, abiotic stress tolerance, and altered metabolism. In other embodiments, the desired trait is herbicide tolerance and the herbicide is selected from, but not limited to, the group consisting of glyphosate, glufosinate, imidazolinones, and auxin analogues such as 2,4-D and dicamba.

[0373] In yet another aspect, the invention is directed to a method of producing a Brassica carinata variety having a desired trait, as well as seeds, plants and plant parts of such variety, by crossing a plant of Brassica carinata variety DH-056.149 with a plant of another species of the family Brassicaceae comprising the desired trait, producing F1 progeny plants using embryo rescue techniques to recover viable F1 plants or growing F1 seeds, self-pollinating the F1 plants that have the desired trait and carinata character, producing F2 plants using embryo rescue techniques to recover viable F2 plants or growing F2 seeds, self-pollinating the F2 plants that have the desired trait and carinata character, using embryo rescue techniques to recover viable F3 plants or growing F3 seeds to produce progeny plants, self-pollinating the progeny plants that have the desired trait and carinata character to produce further progeny plants, and selecting the progeny plants with the desired trait and carinata character. In some embodiments, the steps of producing progeny plants, self-pollinating, and selecting progeny plants having the desired trait and carinata character are repeated until the progeny plant has the desired trait and the physiological and/or morphological characteristics set forth in one or more of Tables 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14, as determined at the 5% significance level, when grown in the same location under the same environmental conditions as variety DH-056.149. In other embodiments, the steps of backcrossing and growing the resulting progeny seed may be repeated 1 to 2 times, 1 to 3 times, 1 to 4 times, or 1 to 5 times, or until the Brassica carinata variety produced from Brassica carinata variety DH-056.149 has the desired trait and the physiological and/or morphological characteristics set forth in one or more of Tables 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14, as determined at the 5% significance level, when grown in the same location under the same environmental conditions as variety DH-056.149. In some embodiments, the desired trait is selected from the group consisting of male sterility, disease resistance, fungal resistance, pest resistance, herbicide tolerance, abiotic stress tolerance, and altered metabolism. In other embodiments, the desired trait is herbicide tolerance and the herbicide is selected from, but not limited to, the group consisting of glyphosate, glufosinate, imidazolinones, and auxin analogues such as 2,4-D and dicamba.

[0374] In another aspect, the present invention is directed to a cell of a plant of a Brassica carinata variety having a desired trait, as well as seeds, plants and plant parts of such variety, by crossing a plant of Brassica carinata DH-056.149 with a plant of another species of the family Brassicaceae comprising the desired trait, producing F1 progeny plants using embryo rescue techniques to recover viable F1 plants or growing F1 seeds, self-pollinating the F1 plants that have the desired trait and carinata character, producing F2 plants using embryo rescue techniques to recover viable F2 plants or growing F2 seeds, self-pollinating the F2 plants that have the desired trait and carinata character, using embryo rescue techniques to recover viable F3 plants or growing F3 seeds to produce progeny plants, self-pollinating the progeny plants that have the desired trait and carinata character to produce further progeny plants, and selecting the progeny plants with the desired trait and carinata character. In some embodiments, the steps of producing progeny plants, self-pollinating, and selecting progeny plants having the desired trait and carinata character are repeated until the progeny plant has the desired trait and the physiological and/or morphological characteristics set forth in one or more of Tables 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14, as determined at the 5% significance level, when grown in the same location under the same environmental conditions as variety DH-056.149. In some embodiments, the desired trait is selected from the group consisting of male sterility, disease resistance, fungal resistance, pest resistance, herbicide tolerance, abiotic stress tolerance, and altered metabolism. In other embodiments, the desired trait is herbicide tolerance and the herbicide is selected from, but not limited to, the group consisting of glyphosate, glufosinate, imidazolinones, and auxin analogues such as 2,4-D and dicamba.

[0375] Another aspect of the present invention is directed to a tissue culture of protoplasts or regenerable cells of the plant or plant part produced from the seed of Brassica carinata variety DH-056.149, as well as a Brassica carinata plant regenerated from the tissue culture the physiological and/or morphological characteristics set forth in one or more of Tables 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14, as determined at the 5% significance level, when grown in the same location under the same environmental conditions as variety DH-056.149. The tissue may be selected from the group consisting of leaves, pollen, embryos, roots, root tips, pods, flowers, ovules, and stalks.

[0376] In other aspects, the present invention is directed to a cell of Brassica carinata variety designated DH-056.149, as well as to a plant or plant part, or a protoplast of a plant or plant part, produced from a seed of Brassica carinata variety designated DH-056.149. In some embodiments, the plant part is an ovule, a leaf, pollen, a seed, an embryo a root, a root tip, a pod, a flower, or a stalk. In other embodiments, the plant part is pollen or an ovule. In other embodiments, the present invention is directed to a cell, plant or plant part having the physiological and/or morphological characteristics set forth in one or more of Tables 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14, as determined at the 5% significance level, when grown in the same location under the same environmental conditions as variety DH-056.149.

[0377] The present invention also includes a method of producing a Brassica carinata variety comprising a desired trait, as well as a plant, plant part, or seed of such variety, by introducing a nucleic acid construct conferring the desired trait into a Brassica carinata plant of variety DH-056.149 using polyethylene glycol (PEG) mediated uptake, electroporation, ballistic infiltration using DNA coated microprojectiles (gene gun), an Agrobacterium infiltration-based vector, or a plant virus-based vector. In some embodiments, the nucleic acid construct comprises a transgene. In other embodiments, the nucleic acid construct comprises an RNAi construct. In some embodiments, the Brassica carinata variety comprises the desired trait and the physiological and/or morphological characteristics set forth in one or more of Tables 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, and 14, as determined at the 5% significance level, when grown in the same location under the same environmental conditions as variety DH-056.149. In other embodiments, the desired trait is selected from the group consisting of male sterility, disease resistance, fungal resistance, pest resistance, herbicide tolerance, abiotic stress tolerance, and altered metabolism. In other embodiments, the desired trait is herbicide tolerance and the herbicide is selected from, but not limited to, the group consisting of glyphosate, glufosinate, imidazolinones, and auxin analogues such as 2,4-D and dicamba.

[0378] In another aspect, the invention is directed to the use of a plant of Brassica carinata variety DH-056.149 to produce a Brassica carinata variety comprising a new trait, wherein the new trait is introduced by exposing seedlings or microspores to a mutagenic agent. In some embodiments, the mutagenic agent is ethyl methanesulfonate, N-ethyl-N-nitrosourea, or x-ray, gamma or ultraviolet radiation.

[0379] In another aspect, the invention is directed to a cell of a plant of a Brassica carinata variety produced from variety DH-056.149, wherein the Brassica carinata variety comprises a new trait and is produced by a method comprising exposing seedlings or microspores to a mutagenic agent and allowing the surviving fraction to develop into mature plants. In some embodiments, the mutagenic agent is ethyl methanesulfonate, N-ethyl-N-nitrosourea, or x-ray, gamma or ultraviolet radiation.

[0380] In another aspect, the invention is directed to a method of producing a commercial crop of a Brassica carinata variety DH-056.149 or of a Brassica carinata variety produced by any of methods described herein. In another aspect, the invention is directed to a method of producing a commercial plant product from the commercial crop. In some embodiments, the commercial plant product comprises oil, meal, protein isolate, or biofumigant. In another aspect the invention is directed to a method of producing crushed, non-viable seed of Brassica carinata variety DH-056.149 or a Brassica carinata variety produced by any of methods described herein.

[0381] In another aspect, the invention is directed to a method of biofumigation comprising, growing a plant of Brassica carinata variety DH-056.149 or a Brassica carinata variety produced by any of methods described herein in a field, collecting the Brassica carinata plant biomass between flowering and seed set, chopping the biomass, and incorporating the chopped biomass into the soil.

[0382] The citation of any publication herein is not an admission that the publication is prior art with respect to the present application.

[0383] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it is readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the scope of the appended claims.

[0384] It is to be understood that any numerical value inherently contains certain errors necessarily resulting from the standard deviation found in the respective testing measurements. Also, as used herein, the term about generally means within 10%, 5%, 1%, or 0.5% of a given value or range. Alternatively, the term about means within an acceptable standard error of the mean when considered by one of ordinary skill in the art. Unless indicated to the contrary, the numerical parameters set forth in the present disclosure and attached claims are approximations that can vary as desired. At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

[0385] As used in this specification and the appended claims, the singular forms a, an, and the include plural reference unless the context clearly dictates otherwise. Unless defined otherwise all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this invention belongs.

[0386] The phrase and/or, as used herein in the specification and in the claims, should be understood to mean either or both of the elements so conjoined, i.e., elements that are conjunctively present in some cases and disjunctively present in other cases. Multiple elements listed with and/or should be construed in the same fashion, i.e., one or more of the elements so conjoined. Other elements may optionally be present other than the elements specifically identified by the and/or clause, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, a reference to A and/or B, when used in conjunction with open-ended language such as comprising can refer, in one embodiment, to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc.

[0387] As used herein in the specification and in the claims, or should be understood to encompass the same meaning as and/or as defined above. For example, when separating items in a list, or or and/or shall be interpreted as being inclusive, i.e., the inclusion of at least one, but also including more than one, of a number or list of elements, and, optionally, additional unlisted items.

[0388] As used herein, whether in the specification or the appended claims, the transitional terms comprising, including, carrying, having, containing, involving, and the like are to be understood as being inclusive or open-ended (i.e., to mean including but not limited to), and they do not exclude unrecited elements, materials or method steps. Only the transitional phrases consisting of and consisting essentially of, respectively, are closed or semi-closed transitional phrases with respect to claims and exemplary embodiment paragraphs herein. The transitional phrase consisting of excludes any element, step, or ingredient which is not specifically recited. The transitional phrase consisting essentially of limits the scope to the specified elements, materials or steps and to those that do not materially affect the basic characteristic(s) of the invention disclosed and/or claimed herein.

EXAMPLES

Example 1: Field Trials

[0389] Small plot field experiments were conducted in Canada and northern states of the United States and Canada using best agronomic practices for carinata as described in the Carinata Management Handbook for the Northern Plains (http://growcarinata.com/ckfinder/userfiles/files/US%20Northern%20Plains_handbook_2018.pdf). Several newly developed carinata varieties, including DH-056.149, were grown in a Randomized Complete Block Design trial (RCBD), comprising 4 replicate plots (unless otherwise noted) along with plots of check varieties AAC-A120 and/or AGRO44-312D-HP11 (henceforth referred to as AGRO44-HP11 or HP11). During the trials, the experimental lines and check varieties were closely observed, and descriptions of phenotypic characteristics and agronomic traits were recorded. After plants had reached maturity, seed was collected for subsequent NIR based seed quality analysis (as described in the definitions section). Least square mean values from replicated data sets were calculated using the REML model and pairwise comparisons carried out using Tukey's test at the 5% probability level. All quantitative data analysis was carried out using JMP 13 (SAS).

Example 2: Varietal Characteristics of DH-056.149 Grown in Southeastern United States

[0390] The southeastern states of Florida, Georgia, Alabama and others constitute an important carinata growing region. Farmers in this region would benefit from a crop that can be grown during the winter months as a cover crop, replacing the normal winter fallow in their rotations, allowing them an additional cash crop option while providing soil benefits that may in turn benefit other crops in their rotation. In order to identify varieties with suitable productivity and agronomic properties in the low daylight hour/winter cropping scenario, field evaluations of new carinata varieties in this region are carried out, selecting the best adapted varieties for the local environment.

[0391] As described in Example 1, replicated field trials afford the opportunity to compare experimental varieties in terms of their morphological phenotypes and agronomic traits. During the winters of 2016-2017 and 2017-18, trials were carried out in Quincy, Fla. and provided extensive phenotypic observations, summarized in Table 1. Each characteristic was assessed as described in DEFINITIONS section, above, with the following exceptions and additions: [0392] Sclerotinia incidence: expressed as a % of symptomatic plants in a 3948 area (data collected 2-4 days before harvest) [0393] Shattering loss at harvestshattered seeds at harvest collected in three 713 pans placed 3 from either end or at center of each plot and converted to kg/ha.

[0394] Table 1 provides data for morphological and agronomic characteristics for variety DH-056.149 and check variety AGRO44-HP11 (HP11). Data were obtained from observations taken during trials carried out Quincy, F1. Least square mean values from replicated data sets were calculated using the REML model and pairwise comparisons carried out using Tukey's test at the 5% probability level. All quantitative data analysis was carried out using IMP 13 (SAS). Paired LSM values that share a common letter (letters column) are not significantly different from one another.

TABLE-US-00002 TABLE 1 Morphological traits of DH-056.149 grown in Quincy, FL 2016-17 and 2017-18 Botanical name Brassica carinata Season type Spring/winter Pollination control Open pollinated Year 2016-17 2017-18 Variety DH-056.149 Letters HP11 Letters DH-056.149 Letters HP11 Letters Days to first 80.5 DE 84.25 BCD ND* ND bolt Days to 50% 88.0 E 89.5 CDE 94.0 AB 94.5 AB bolt Days to first 90.0 C 93.5 B ND* ND* ND* flower Days to 50% 100.0 C 105.5 B 101.25 BC 103.5 AB flower Days to last 127.25 CD 135.50 A Nd* ND* Nd* flower Days to 166.75 B 169.0 A 161.8 BCDE 162.2 BCDE Maturity Depth of canopy 71.63 A 79.25 A (cm) Stand 39.75 A 42.44 A ND* ND* (plants/m2) Plant height 152 D 184.4 A ND* Nd* (cm) Pod shatter 316.9 A 189.6 ABCD ND* Nd* loss (kg/ha) ScLerotinia 22.57 AB 8.25 CD ND* Nd* incidence, % TKW (g) 5.10 ABC 4.79 BCDE 3.29 F 4.33 ABC ND* = not determined

[0395] The distinguishing characteristics of DH-056.149 with respect to the check variety are summarized below

TABLE-US-00003 Flowering: In both trial years, plants of DH-056.149 bolted and/or initiated flowering earlier than plants of the check variety. The end of flowering was also earlier for DH-056.149 in the 2016-17 trial. Plant In the 2016-17 trials, plants of DH-056.149 were found to be height: significantly shorter in stature than plants of the check variety; plants of DH-056.149 also had a shorter canopy than the check variety. Maturity: In both trial years, maturity occurred slightly earlier for plants of DH-056.149 compared to plants of the check variety; however, this difference was only found to be significant in the 2016-17 trial. Pod Loss of grain due to pod shattering, in kg/ha, was not Shatter statistically different for variety DH-056.149 compared to Loss check variety AGR044-HP11. Sclerotinia Under the growth conditions of this trial, DH-056.149 showed suscep- a significantly higher incidence of Sclerotinia infection tibility: (22.6%) than check variety AGR044-HP11.

[0396] In summary, variety DH-056.149 exhibits a number of agronomic traits consistent with those of an existing commercial variety grown in the SE US region, as well as other traits (early flowering and maturity) that would make it a suitable candidate for commercial cultivation in the SE USA or as a breeding line.

Example 3: Grain Yields of DH-056.149 Grown in Winter Trials in Southeast US States

[0397] To assess the yield potential of new and experimental carinata varieties, including DH-056.149, relative to commercial check varieties, a series replicated yield trials designed as described in Example 1 were carried out during winter 2016-2017 and 2017-18 in southeast US. Trials were carried out at four sites. However, all but one of the sites (Quincy, F1) demonstrated coefficients of variation (C of V) for yields too high (>15%) to be considered reliable; therefore, only yield data from Quincy were considered.

[0398] After maturity had been reached, seeds were harvested from the center 5 of each plot (5 rows), cleaned and dried in a forced air oven at 50 C. for at least 72 h, subsequently weighed for yield determination and the seed analyzed for test weight and moisture using a Steinlite SL95 Moisture Meter. Reported yields, tabulated in Table 2, have been adjusted to 8% moisture. Paired LSM values that share a common letter (letters column of Table 2) are not significantly different from one another.

TABLE-US-00004 TABLE 2 Grain yields of variety DH-056.149 grown in Quincy, Florida 2016-17 2017-18 LSM Yield LSM Yield Variety (kg/ha) Letters (kg/ha) Letters DH-056.149 2673 A 2608 ABC AGR044-HP11 2106 AB 3114 A

[0399] The LSM yield of DH-056.149 is statistically similar to the yield of the AGRO44-HP11 check variety (Table 2), making it a suitable candidate for commercial exploitation in the southeast region of the US.

Example 4: Quality of Carinata Seed Harvested from Winter Field Trials in Southeast US

[0400] Trials were conducted in southeastern US states as described in Example 1. Seed quality data were obtained from NIR analysis, as described above under DEFINITIONS, on replicated samples of harvested grain from small plot, RCBD field experiments. Least square means (LSM) were calculated for measurements of replicate samples using a restricted maximal likelihood (REML) estimate, which can better describe data sets where some replicate data might be missing due to weather or disease related losses. Tukey's test was then applied pairwise to determine whether there were statistically significant differences between the LSM yields for DH-056.149 and the check variety (Table 3). Paired LSM values that share a common letter (letters column of Table 3) are not significantly different from one another.

TABLE-US-00005 TABLE 3 Seed quality characteristics of variety DH-056.149 grown in southeast US in 2016-17 (Citra and Quincy, FL) and 2017-18 (Quincy, FL) 2016-17 2017=18 Quality Variety LSM Letters LSM Letters Seed Protein DH-056.149 24.86 EF 27.91 BC content, wt % of AGR044-HP11 25.07 EF 25.50 DE seed Seed Oil content, DH-056.149 46.35 BC 49.00 A wt % of seed AGR044-HP11 45.84 C 47.91 ABC Glucosinolates, DH-056.149 93.37 BC 106.01 ABC mol/g AGR044-HP11 83.59 C 93.26 D Erucic Acid, DH-056.149 37.57 G 47.80 AB wt % of total AGR044-HP11 44.17 B 46.05 BCD fatty acids Total Saturates, DH-056.149 5.87 AB 5.04 F wt % of total AGR044-HP11 5.51 CD 5.75 DE fatty acids

[0401] The fatty acid profile of seed oil from DH-056.149, as determined by NIR analysis, was also compared to that of seed from check variety AGRO44-HP11 and the results, expressed as the percentage by weight (mean of 4 samples) for each of the major fatty acid constituents of the oil is shown in Table 4. Paired LSM values that share a common letter (letters columns) are not significantly different from one another.

TABLE-US-00006 TABLE 4 Fatty acid profile of oil from DH-056.149 grown in Quincy, Fl in 2017-18 Fatty Acid Variety LSM Letters C18:1 DH-056.149 0.56 E AGR044-HP11 5.13 CD C18:2 DH-056.149 12.70 D AGR044-HP11 14.51 ABC C18:3 DH-056.149 10.60 G AGR044-HP11 11.96 CD C20:1 DH-056.149 11.38 A AGR044-HP11 9.37 CDE C22:1 DH-056.149 47.80 AB AGR044-HP11 46.05 BCD Mono DH-056.149 62.84 BCDE AGR044-HP11 63.72 BC Poly DH-056.149 29.22 EF AGR044-HP11 29.94 CDE LCFA DH-056.149 27.21 F AGR044-HP11 32.28 DE VLCFA DH-056.149 72.79 A AGR044-HP11 67.72 BC ND = not determined

[0402] The results of the seed quality analysis of DH-056.149 grown in southeast US during the winters of 2016-2017 and 2017-18 are summarized below can be summarized as follows:

TABLE-US-00007 Seed oil content Seed from DH-056.149 has, on average, a statistically similar oil content as seed from check variety AGR044-HP11. Seed protein content Seed from DH-056.149 has, on average, a statistically similar (2016-17) or somewhat lower (2017-18) protein content as seed from check variety AGR044-HP11. Glucosinolates The total GSL content (mol/g) of seed produced by DH- 056.149 is, on average, statistically similar to (2016-17) or somewhat higher than (2017-18) the total GSL content of seed produced by the check variety AGR044-HP11. Erucic acid Oil from DH-056.149 grain produced in the 2016-17 trials shows significantly lower levels of erucic acid (C22:1) compared to oil from grain of check variety AGR044-HP11; however, in the 2017-18 trials, the erucic acid content was similar in the oil from grain produced by both DH-056.149 and the check variety. Total saturates Oil from DH-056.149 grain shows statistically significant higher (2016-17) or lower (2017-18) levels of total saturates compared to oil from grain of check variety AGR044-HP11. Fatty acid profile Oil from DH-056.149 grain produced in the 2017-18 trial was enriched in the very long chain fatty acids (VLCFA) such as eicosenoic acid (C20:1) and erucic acid (C22:1) compared to oil from grain produced by the check variety, and had a corresponding and significant lower proportion of the long chain fatty acids (LCFA) such as oleic (C18:1), linoleic (C18:2) and linolenic (C18:3) acids.

[0403] As can be seen in Table 3, the DH-056.149 showed a similar oil content to that of the commercial check variety which, based on yield equivalence between DH-056.149 and the check line (shown in Example 3), could result in similar amounts of oil recovered per hectare for fields planted with DH-056.149 and current commercial variety. Although the oil in DH-056.149 grain harvested in this trial had a slightly lower erucic acid content that of the check variety, at almost 38 wt % it was still within the typical range seen for current commercial carinata varieties.

[0404] In these trials, DH-056.149 produced seed with a highly significant 23 mol/g lower total GSL content than the commercial check line. The significantly lower levels of GSL found in the DH-056.149 seed could positively impact the quality of oil and meal feedstocks produced from its grain. Even lower GSL inclusion rates in carinata meal may be achieved through application of downstream processing technologies, such as that described in U.S. Publication No. US2018/0042266 A1.

[0405] DH-056.149 seed produced in this trial contained a similar protein content as that of the check line. Therefore, meal produced from processing of this seed would still be expected to contain >40% protein content, representing an advantage with respect to competing Brassica oil seed meals such as canola meal.

[0406] Based on the potential for oil yields similar to oil yields from existing commercial varieties as well as the seed quality attributes discussed above, the DH-056.149 variety may be a suitable candidate for commercial exploitation in the southeast region of the US.

Example 5: Varietal Characteristics of DH-056.149 Grown in Field Trials in Northern US and Canada

[0407] The northern tier states of the US, such as North Dakota and its neighbours, as well as the adjacent southern Canadian prairie regions of Alberta and Saskatchewan constitute an important Brassica oilseed growing region. Evaluation of new carinata varieties in this region allows for identification of those varieties that are best adapted to growing in the local environment. which comprises a semiarid environment with warm summer and cold winter season. Under such conditions, carinata varieties which are productive under long daylight spring-summer cultivation in semiarid environments would prove advantageous to local farmers. Field trials are carried out annually at several sites in the region as described in Example 1 to identify best candidates for commercialization based on agronomic traits, seed yield and quality as well as multi-year performance.

[0408] The detailed phenotypic information provided in Table 5 is based on data collected from small plot, RCBD field experiments conducted in northern US states and Canada during the spring and summer, as described in Example 1. Each characteristic was assessed as described in DEFINITIONS section, for all experiment carinata varieties entered in the trial, including DH-056.149, as well as the commercial check variety AAC-A120, a carinata variety well adapted to cultivation in the target region. For all values from the 2018 trials, as well as for the values from the 2017 trial for days to flower, days to maturity, and pod canopy, measurements represent least square mean of replicated ratings or measurements taken at 4 or more of the sites. For the 2017 trials, the values for early vigour represent averages of ratings assigned at 4 or more sites, the values for maturity average ratings from three or more sites, and the values for lodging resistance average ratings from one or two sites.

[0409] Each characteristic was assessed as described in DEFINITIONS section, above. Table 5 provides additional data for morphological and agronomic characteristics for variety DH-056.149. and check variety AAC-A120.

TABLE-US-00008 TABLE 5 Morphological traits of DH-056.149 grown in field trials in northern US and Canada Botanical name Brassica carinata Season type Spring/winter Pollination control Open pollinated Year 2017 2018 Variety DH-056.149 Letters AAC-A120 Letters DH-056.149 Letters AAC-A120 Letters Early vigour 5.53 A 5.33 A ND ND (1-7 scale) Days to first 51.25 B 48.62 CD 56.75 A 56.00 A flower End flowering 79.8 77.3 71.00 AB 74.50 A (days) Flowering 31.5 31.3 14.25 A 18.50 A duration (days) Days to maturity 109.0 A 106.4 A Bottom pod 51.10 A 46.60 AB 42.5 A 28.75 A canopy (cm) Canopy height 106.9 A 96.73 AB 100.5 A 89.75 AB (cm) Depth of canopy 56.53 A 53.14 AB 58.00 AB 61.00 ABC (cm) Lodging 5.5 5.1 1.5 B 6.0 A resistance (1-7 scale) Shatter 4.0 4.8 5.5 AB 3.5 B resistance (1-7 scale) ND = not determined.

[0410] Relative to check variety AAC-A120, Brassica carinata variety DH-056.149 shows the following agronomic characteristics:

TABLE-US-00009 Flowering In both trial years, some differences were observed between plants of variety DH-056.149 and plants of the check variety with regard to initiation of flower, end of flowering, or duration of flowering; however, these differences were not found to be statistically significant. Canopy In both trial years, some differences were observed between plants of variety DH-056.149 and plants of the check variety with regard to canopy height, bottom of canopy and canopy depth; however, these differences were not found to be statistically significant. Maturity In the 2017 trial, plants of variety DH-056.149 reached maturity 2-3 days later than plants of check variety AAC-A120. Lodging In the 2018 trial, plants of variety DH-056.149 a significantly higher propensity to lodging compared to the check variety AAC-A120. Resistance to Pod Resistance to pod shattering varied from year to year, with pods of variety DH-056.149 exhibiting slightly lower (2017) or higher Shattering (2018) resistance to pod shattering compared to pods of check variety AAC-A120.

[0411] Agronomic data accumulated from the trials described above indicate that variety DH-056.149 was found to possess traits consistent with those of an existing commercial line typically grown in the northern US and Canadian southern prairies and supports selection of variety DH-056.149 as suitable candidate for commercial cultivation in this region.

Example 6: Grain Yields of Variety DH-056.149 from Trials in Northern US and Canada

[0412] Tables 6, 7, and 8 (below) summarize the results of yield analysis carried out on DH-056.149 and check variety AAC-A120 from field trials in northern US and Canada in 2016, 2017 and 2018. Least square means (LSM) were calculated for yields from replicate trials for each variety tested at each site using a restricted maximal likelihood (REML) estimate, which can better describe data sets where some replicate data might be missing due to weather or disease related losses. Tukey's test was then applied pairwise to determine whether there were statistically significant differences between the LSM yields for DH-056.149 and check variety AAC-A120. An aggregate of the yield data from all sites was obtained and the aggregated LSM yield values for each variety were compared to one another as described above. Paired LSM values that share a common letter (letters columns of Tables 6, 7, and 8) are not significantly different from one another.

TABLE-US-00010 TABLE 6 Grain yields of DH-056.149 from 2016 trials in northern US states and Canada LSM Yield Site Variety (kg/ha) Letters Medicine Hat, AB DH-056.149 1851 ABCD AAC-A120 1806 ABCD Devils Lake, ND DH-056.149 3590 A AAC-A120 3136 A Moosomin SK DH-056.149 3018 ABCD AAC-A120 2704 ABCDE Hettinger, ND DH-056.149 1369 A AAC-A120 1077 AB All sites DH-056.149 2897 ABC AAC-A120 2547 ABCD

TABLE-US-00011 TABLE 7 Grain yields of DH-056.149 from 2017 trials in northern US states and Canada LSM Yield Site Variety (kg/ha) Letters Carrington, ND DH-056.149 2581 AB AAC-A120 2086 BC Indian Head, SK DH-056.149 3447 A AAC-A120 3057 A Milestone, SK DH-056.149 1830 AB AAC-A120 1827 AB Outlook, SK DH-056.149 3459 A AAC-A120 3434 A All sites DH-056.149 2837 A AAC-A120 2601 AB

TABLE-US-00012 TABLE 8 Grain yields of DH-056.149 from 2018 trials in northern US states and Canada LSM Yield Site Variety (kg/ha) Letters Brookings, ND DH-056.149 1576 AB AAC-A120 1849 A Outlook, SK DH-056.149 3074 A AAC-A120 3732 A

[0413] While the yield data reflect considerable variation between sites, at each individual site the difference in LSM yields between DH-056.149 and the commercial check variety AAC-A120 were found not to be significantly different.

[0414] The data from trials carried out in northern US states and Canadian southern prairies in 2016 (Table 6), 2017 (Table 7), and 2018 (Table 8) demonstrate similar yield potentials for DH-056.149 and a commercial carinata variety used as check variety, supporting selection of DH-056.149 as a candidate for commercial cultivation in this region.

Example 7: Quality of Carinata Seed Harvested from Field Trials Conducted in Northern US and Canada

[0415] Trials were conducted as described in Example 1. Seed quality data were obtained from NIR analysis, as described above under DEFINITIONS, on replicated samples of harvested grain from small plot, RCBD field experiments. Least square means (LSM) were calculated for measurements of replicate samples using a restricted maximal likelihood (REML) estimate, which can better describe data sets where some replicate data might be missing due to weather or disease related losses. Tukey's test was then applied pairwise to determine whether there were statistically significant differences between the LSM values for DH-056.149 and check variety AAC-A120 (Table 9). Paired LSM values that share a common letter (letters column of Table 9) are not significantly different from one another.

TABLE-US-00013 TABLE 9 Seed quality characteristics of DH-056.149 from trials in northern US and Canada. 2016 2017 2018 Quality Variety LSM Letters LSM Letters LSM Letters Protein content, DH-056.149 29.25 BC 27.5 DE 30.41 A wt % of seed AAC-A120 29.35 BC 28.5 ABCD 30.36 AB Oil content, DH-056.149 40.58 E 43.2 ABCD 39.68 E wt % of seed AAC-A120 43.12 BC 44.3 A 42.81 D Glucosinolates, DH-056.149 107.12 AB 91.4 ABC 86.16 AB mol/g AAC-A120 102.88 B 80.6 ABC 78.16 ABC Erucic Acid, DH-056.149 40.20 E 36.9 JK 40.40 C wt % of fatty acids AAC-A120 43.04 CD 40.5 CDE 43.62 BC Total Saturates, DH-056.149 6.12 A 6.26 A wt % of fatty acids AAC-A120 5.71 EF 5.80 CD

[0416] The fatty acid profile of seed oil from DH-056.149, as determined by NIR analysis, was also compared to that of the AAC A120 check variety and the results, expressed as the percentage by weight (mean of 4 samples) for each of the major fatty acid constituents of the oil are shown in Table 10. Paired LSM values that share a common letter (letters column of Table 10) are not significantly different from one another.

TABLE-US-00014 TABLE 10 Fatty acid profile of DH-056.149 grown in northern US states and Canadian southern prairies in 2018 Fatty Acid Variety LSM Letters C18:1 DH-056.149 8.65 A AAC-A120 6.28 C C18:2 DH-056.149 16.91 A AAC-A120 15.29 CD C18:3 DH-056.149 12.61 E AAC-A120 14.40 C C20:1 DH-056.149 9.08 ABC AAC-A120 8.97 ABCD C22:1 DH-056.149 40.40 C AAC-A120 43.62 BC Mono DH-056.149 58.70 DE AAC-A120 58.02 E Poly DH-056.149 33.94 AB AAC-A120 34.20 A LCFA DH-056.149 40.77 AB AAC-A120 39.70 AB VLCFA DH-056.149 59.23 DE AAC-A120 60.30 DE

TABLE-US-00015 Seed oil content Grain from DH-056.149 exhibited statistically similar (2017), or somewhat lower (2016 and 2018), oil content relative to the oil content of grain from the check variety. Seed protein content In all trial years, grain from DH-056.149 exhibited statistically similar protein content as grain from the check variety. Glucosinolates In all trial years, grain from DH-056.149 exhibited statistically similar total glucosinolate content as grain from the check variety. Erucic acid In the 2016 and 2017 trials, oil from DH-056.149 grain contained statistically lower levels of erucic acid (~3 wt %) compared to oil from grain of the check variety; however, in the 2018 trials, the erucic acid content of the oil from grain of DH-056.149 was similar to that from grain of the check variety. Total saturates and Oil from grain of DH-056.149 produced in the 2018 trial had a fatty acid profile significantly higher proportion of saturated fatty acids, as well as higher proportion of the three C18 fatty acids (oleic acid, C18:1, linoleic acid, C18:2, and linolenic acid, C18:3).

[0417] As can be seen in Table 9, grain from DH-056.149 showed a similar, or slightly lower, oil content compare to grain from the commercial check variety which, based on equivalent or slightly higher yields for variety DH-056.149 compared to the check variety (shown in Example 6), could result in similar amounts of oil recovered per hectare for fields planted with DH-056.149 and current commercial varieties.

[0418] In these trials, DH-056.149 produced grain with a somewhat, though not significantly, higher total GSL content than the commercial check variety.

[0419] DH-056.149 grain produced in this trial contained a similar protein content as that of the check variety. Meal produced from processing of this grain would therefore be expected to contain >40 wt % protein content, representing an advantage with respect to competing Brassica oil seed meals such as canola meal.

[0420] Based on the potential for oil yields equivalent those from the existing commercial variety, as well as the seed quality attributes discussed above, the DH-056.149 variety may be a suitable candidate for commercial exploitation in the northern US states and Canada, and possibly other geographical regions.

Example 8: Varietal Characteristics of DH-056.149 when Grown in South America

[0421] Uruguay is an important commercial producer of Brassica carinata. Its climate lends itself to production of carinata as a short-day length winter cover crop often in rotation with summer grown crops such as soybean. In order to assess, new carinata varieties for their suitability in commercial production in this region, small plot yield trials have been carried out in Uruguay, primarily at two sites, La Estanzuela (LE) and Young (YO). In winter of 2018, small plot trials were carried out at both sites, essentially as described in Example 1, except that 3 randomized plot replicates were employed instead of 4 and provided extensive phenotypic observations, summarized in Table 11. Each characteristic was assessed as described in DEFINITIONS section, above.

TABLE-US-00016 TABLE 11 Morphological traits of variety DH-056.149 grown in field trials carried out in Uruguay (2018) DH-056.149 AGR044-HP11 Traits LE YO LE YO 10% flowering (days) 113 85 110 85 Mid flowering (days) 119 88 118 88 100% flowering (days) 140 108 157 116 Plant height (m) 1.86 1.66 1.83 1.55 Canopy bottom (m) 0.96 0.91 0.73 0.65 Canopy depth (m) 0.90 0.75 1.10 0.90 Stand (plants/m.sup.2) 63 ND 59 ND Shatter (kg/ha) 205 ND 170 ND Harvest loss (kg/ha) 415 ND 335 ND TKW (g) 3.0 ND 3.7 ND ND = not determined

[0422] The distinguishing characteristics of DH-056.149 grown in this trial and with respect to the check variety are summarized below:

TABLE-US-00017 Flowering: Plants of variety DH-056.149 initiated flowering within 3 days of plants of the check variety; however, plants of variety DH-056.149 ended flowering 1-2 weeks earlier than plants of the check variety. Plant height: Plants of variety DH-056.149 were similar in height or slightly taller than plants of the check variety; however, the canopy bottom was observed to be higher for the DH-056.149 compared to plants of the check variety at both trial sites. As a result, plants of DH-056.149 exhibited a small depth of canopy compared to plants of the check variety. Stand: Plant stand determinations (number plants/m.sup.2 prior to harvest), carried out solely at the La Estanzuela site, were similar for variety DH- 056.149 and the check variety. Pod shatter: Pod shatter, expressed as absolute seed loss (LSM kg of seed/ha) obtained from replicate plots prior to harvest, was determined as described in the definition section at the La Estanzuela site. DH- 056.149 experienced similar harvest losses to the check variety.

Example 9: Yield of Carinata Seed from Variety DH-056.149 Grown in Uruguay During Winter

[0423] Table 12 summarizes the results of yield analysis carried out on a number of new and experimental carinata varieties at La Estanzuela in 2018. Several new varieties were evaluated, including DH-056.149, alongside check variety AGRO44-HP11, the latter chosen due to it being the current preferred commercial variety in this geography. Least square means (LSM) yield (in kg/ha) were calculated from replicate trials for each variety tested at each site using a restricted maximal likelihood (REML) estimate, which can better describe data sets where some replicate data might be missing due to weather or disease related losses. Tukey's test was then applied pairwise to determine whether there were statistically significant differences between the LSM yields for DH-056.149 and check variety AGRO44-HP11. Paired LSM values that share a common letter (letters column of Table 12) are not significantly different from one another.

TABLE-US-00018 TABLE 12 Yield of seed from variety DH-056.149 grown in Uruguay during winter 2018 LSM Yield Site Variety (kg/ha) Letters La DH-056.149 3253 AB Estanzuela AGR044-HP11 4110 AB ND = not determined

[0424] As shown Table 12, analysis of LSM yield values by Tukey's test showed that there were no statistically significant differences between yields observed for DH-056.149 and the check variety.

Example 10: Quality of DH-056.149 Carinata Seed Harvested from Field Trials Conducted in Uruguay

[0425] As described above, and in Example 1, trials were conducted using best agronomic practices for carinata as describe the Carinata Management Handbook for the southeastern United States (https://argisoma.com/ckfiner/userfiles/files/2017_18_SE_Handbook.pdf) Seed quality data was obtained from NIR analysis of replicated samples of harvested grain, as described under DEFINITIONS, above. Table 13 tabulates the main characteristics of DH-056.149 seed harvested from these trials in relation to those of the HP11 check variety. Paired LSM values that share a common letter (letters columns of Tables 13 and 14) are not significantly different from one another.

TABLE-US-00019 TABLE 13 Seed quality characteristics of variety DH-056.149 grown in Uruguay during winter of 2018 Quality Variety LSM Letters Protein content, DH-056.149 27.74 AB wt % of seed HP11 26.26 ABCD Oil content, DH-056.149 43.14 D wt % of seed HP11 46.6 BC Glucosinolates, DH-056.149 67.55 A mol/g HP11 55.28 ABC Erucic Acid, DH-056.149 40.57 E wt % fatty acids HP11 46.53 AB Total Saturates, DH-056.149 6.46 B wt % fatty acids HP11 6.15 DEF

[0426] The fatty acid profile of seed oil from DH-056.149, as determined by NIR analysis, was also compared to that of the ABR044-HP11 check variety and the results, expressed as the percentage by weight (mean of 4 samples) for each of the major fatty acid constituents of the oil are shown in Table 14.

TABLE-US-00020 TABLE 14 Fatty acid profile of variety DH-056.149 grown in Uruguay during winter of 2018 % Fatty Acid Variety LSM Letters C18:1 DH-056.149 9.57 CD AGR044-HP11 6.28 F C18:2 DH-056.149 16.09 B AGR044-HP11 14.38 C C18:3 DH-056.149 12.70 F AGR044-HP11 14.21 BC C20:1 DH-056.149 8.72 AB AGR044-HP11 7.57 E C22:1 DH-056.149 40.57 E AGR044-HP11 46.53 AB Mono DH-056.149 58.36 EFG AGR044-HP11 59.87 CDE Poly DH-056.149 34.81 CD AGR044-HP11 33.90 DE LCFA DH-056.149 45.17 BC AGR044-HP11 43.62 CD VLCFA DH-056.149 54.83 CD AGR044-HP11 56.38 BC ND = not determined

[0427] The major seed quality trait characteristics of DH-056.149 are summarized below

TABLE-US-00021 Seed oil Seed from DH-056.149 demonstrated a lower oil content content compared to seed from the check variety. Seed protein Seed from DH-056.149 demonstrated similar protein content content as seed from the check variety. Glucosinolates Seed from DH-056.149 demonstrated a higher total GSL content compared to seed from the check variety. Erucic acid Seed from DH-056.149 demonstrated a lower level of erucic (C22.1) compared to oil from seed from the check variety. Fatty acids Oil from seed of DH-056.149 contained higher profile and proportions of saturated fatty acids, oleic acid (C18:1), SATS content and linoleic acid (C18:2) and lower portions of linolenic acid (C18:3) and erucic acid (C22:1). However, the proportions of mono- and polysaturated fatty acids, as well as the proportions of long chain (LCFA) and very long chain (VLCFA) fatty acids were similar to those for oil from seed of the check variety.

[0428] In summary the seed oil content and fatty acid profile, protein content and GSL content of DH-056.149 were found to be substantially comparable to those of an existing commercial variety HP11 grown in the same geography. These attributes support the selection of DH-056.149 as a potential commercial candidate for cultivation in this region

DEPOSIT

[0429] Applicant(s) have made a deposit of at least 2500 seeds of Brassica carinata variety DH-056.149 with the NCIMB Ltd., Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, UK, AB21 9YA, under Accession number 43000. The seeds deposited with NCIMB on Apr. 3, 2018 for DH-056.149 were taken from the seed stock maintained by Agrisoma Biosciences Inc., since prior to the filing date of this application. The deposit of seed of Brassica carinata variety DH-056.149 will be maintained in the NCIMB depository, which is a public depository, for a period of 30 years, or 5 years after the most recent request, or for the enforceable life of the patent, whichever is longer, and will be replaced if it becomes nonviable during that period. Additionally, Applicant has satisfied all the requirements 37 C.F.R. 1.801-1.809, including providing an indication of the viability of the sample upon deposit. Applicant imposes no restrictions on the availability of the deposited material from NCIMB; however, Applicant has no authority to waive any restrictions imposed by law on the transfer of biological material or its transportation in commerce. Applicant(s) do not waive any infringement of their rights granted under this patent or rights applicable to Brassica carinata variety DH-056.149 under the Plant Variety Protection Act (7 USC 2321 et seq.).

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