Broccoli hybrid PS05151639

Abstract

The invention provides seed and plants of broccoli hybrid PS05151639 and the parent lines thereof. The invention thus relates to the plants, seeds and tissue cultures of broccoli hybrid PS05151639 and the parent lines thereof, and to methods for producing a broccoli plant produced by crossing such plants with themselves or with another broccoli plant, such as a plant of another genotype. The invention further relates to seeds and plants produced by such crossing. The invention further relates to parts of such plants.

Claims

1. A broccoli floret from a Brassica oleracea plant, wherein said floret comprises an endogenous content of MSB (glucoraphanin) and MSP (glucoiberin) present from about 2.4 more MSB than broccoli variety Heritage or about 3.5 more MSB than broccoli variety Marathon, when grown in the same environment, wherein the ratio of MSB to MSP in said floret is about 2:1 on a weight to weight basis, and wherein the expression of the endogenous MSB and MSP content shares the genetic source for said content found in broccoli hybrid PS05151639, representative seed of which were deposited under ATCC Accession Number PTA-9676.

2. The floret of claim 1, wherein the floret contains at least about 10 mole/g dry weight MSB and MSP.

3. The floret of claim 1, wherein the floret is obtained from a hybrid broccoli plant.

4. The floret of claim 1, wherein the floret is obtained from a broccoli plant adapted for commercial cultivation.

5. A plant that produces the floret of claim 1.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 shows MSB levels expressed in mg/148 g of starting material for hybrid PS05151639 (labeled PX1639) and two comparison varieties. MSB was quantified in stalks and florets by standard methods.

(2) FIG. 2 shows MSB levels in PS05151639 (1639) and two comparison varieties grown at five different locations in one growing season. Trials were conducted in RCB with 3-4 replicates per variety or in strip plots, unreplicated within a location. MSB was quantified by standard methods.

(3) FIG. 3 shows MSB levels in PS05151639 (1639) and two comparison varieties grown at five different locations in one growing season. Trials were conducted in RCB with 3-4 replicates per variety or in strip plots, unreplicated within a location. MSB was quantified by standard methods.

(4) FIG. 4 shows MSP levels in PS05151639 (1639) and two comparison varieties grown at five different locations in one growing season. Trials were conducted in RCB with 3-4 replicates per variety or in strip plots, unreplicated within a location. MSP was quantified by standard methods.

(5) FIG. 5 shows MSP levels in PS05151639 (1639) and two comparison varieties grown at five different locations in one growing season. Trials were conducted in RCB with 3-4 replicates per variety or in strip plots, unreplicated within a location. MSB was quantified by standard methods.

(6) FIG. 6 shows MSB LSM (least-squared means) analysis for each locationplanting combination shown in FIG. 2. p-values show that PS05151639 (1639) has a significantly higher MSB content than the comparison varieties (p<0.15).

(7) FIG. 7 shows MSB LSM (least-squared means) analysis for each locationplanting combination shown in FIG. 3. p-values show that PS05151639 (1639) has a significantly higher MSB content than the comparison varieties (p<0.10).

(8) FIG. 8 shows MSB times-difference analysis for each locationplanting combination shown in FIG. 2.

(9) FIG. 9 shows MSB times-difference analysis for each locationplanting combination shown in FIG. 3.

(10) FIG. 10 shows a combined analysis of MSB levels across two replicates and multiple locations (data from FIG. 2 and FIG. 3 combined). Data for PS05151639 is labeled 1639.

(11) FIG. 11 shows a combined analysis of MSP levels across two replicates and multiple locations (data from FIG. 2 and FIG. 3 combined). Data for PS05151639 is labeled 1639.

DETAILED DESCRIPTION OF THE INVENTION

(12) The invention provides methods and compositions relating to plants, seeds and derivatives of PS05151639 broccoli hybrid PS05151639 and/or broccoli lines BRM 51-1162 and BRL 51-1128. The parent lines show uniformity and stability within the limits of environmental influence for the traits described hereinafter. By crossing the parent lines, uniform plants of hybrid PS05151639 can be obtained.

(13) In one embodiment, a plant of the invention comprises one or more improved trait selected from increased levels of the phytochemicals MSP (glucoiberin) and/or MSB (glucoraphanin) levels. The development of broccoli hybrid PS05151639 and its parent lines can be summarized as follows.

A. ORIGIN AND BREEDING HISTORY OF BROCCOLI HYBRID PS05151639

(14) The parents of hybrid PS05151639 are BRM 51-1162 and BRL 51-1128. These parents were created as follows.

(15) Line FT-69 is a line developed by the John Innes Center, UK which has elevated levels of the phytochemical MSP (glucoiberin). It was created by crossing a wild relative of domesticated broccoli, Brassica villosa, with a domesticated broccoli, Brassica oleracea. FT-69 was backcrossed to the adapted broccoli parent line BRM 51-19. After each cross, plants were selected based on phenotype similarities to the recurrent parent BRM 51-19, and analyzed for levels of MSP (glucoiberin) and the additional phytochemical MSB (glucoraphanin). The finished line was named BRM 51-1162.

(16) BRL 51-1128 was developed from an original cross of BRL 51-99 single cross (SC) with a cytoplasmic male sterile (CMS) Broccoli having the cabbage Blue Dynasty cytoplasm. BRL 51-99 was the doubled haploid inbred line 398-1254. A back cross program using BRL 51-99 as the recurrent pollen parent for 5 generations was pursued. After five generations of backcrossing, BRL 51-1128 CMS BC 5 line was sent to a production site and the breeder source 05BajaBK 76-2A/CP 1915 was created and used in foundation seed program for this inbred.

B. PHYSIOLOGICAL AND MORPHOLOGICAL CHARACTERISTICS OF BROCCOLI HYBRID PS05151639, BROCCOLI LINE BRM 51-1162 AND BROCCOLI LINE BRL 51-1128

(17) In accordance with one aspect of the present invention, there is provided a plant having the physiological and morphological characteristics of broccoli hybrid PS05151639 and the parent lines thereof. A description of the physiological and morphological characteristics of such plants is presented in Tables 1-3.

(18) TABLE-US-00001 TABLE 1 Physiological and Morphological Characteristics of Hybrid PS05151639 CHARACTERISTIC PS05151639 Heritage Species Brassica oleracea var. italica L. Brassica oleracea var. italica L. Region of Adaptation (area Pacific Coast Pacific Coast where best adapted in USA) Maturity Fall Planted (days from 129.5 129.5 direct seeding to 50% Rep 1 Jan. 8, 2009 rep 2 Jan. 22, 2009 Rep 1 Jan. 8, 2009 rep 2 Jan. 22, 2009 harvest) Rep 1 May 21, 2009 rep 2 May 28, 2009 Rep 1 May 21, 2009 rep 2 May 28, 2009 Fall Planted (days from 80 80 transplanting to 50% Rep 1 Feb. 27, 2009 rep 2 Mar. 12, 2009 Rep 1 Feb. 27, 2009 rep 2 Mar. 12, 2009 harvest) Fall Planted (length of 1 1 harvest period in days) Rep 1 May 21, 2009 rep 2 May 28, 2009 Rep 1 May 21, 2009 rep 2 May 28, 2009 Rep 1 May 21, 2009 rep 2 May 28, 2009 Rep 1 May 21, 2009 rep 2 May 28, 2009 Harvest Season (main Spring/Summer Spring/Summer crop at 50% harvest) Time of Harvest Maturity Early Medium (50% of plants) (Galaxy, Packman, Scorpio) Time of Beginning of Medium Medium Flowering (50% of plants (Coaster, Cruiser) with at least 10% flowers) Seedling Cotyledon Color Medium Green Medium Green Cotyledon Color (RHS 132D 132D color chart value) Cotyledon Anthocyanin Absent Weak Hypocotyl Anthocyanin Intermediate Intermediate Plant Plant Height (at harvest, 68.2 74.9 cm) Head Height (at harvest, 35.9 38.8 cm) Height (at harvest Medium Tall maturity, cm) (Coaster) Number of Stems One One (Ramoso Calabrese, Shogun) Branches Many Medium Habit Intermediate Intermediate Market Class Both Both Life Cycle Annual Annual Type of Variety First Generation Hybrid Hybrid Leaf Width Medium Broad (Buccaneer, Green Belt) Length (including petiole) Long Long (Green Duke, Laser) Number of Lobes Medium Many (Coaster, Topper) Attitude (at beginning of Semi-erect Semi Erect head formation) (Arcadia, Asti, Civet, Claudia) Outer Leaves Number of Leaves Per 16.09 14.3 Plant (at harvest) Width (at midpoint of 16.4 18 plant including petiole, cm) Length (at midpoint of 54.6 60.1 plant including petiole, cm) Petiole Length (cm) 22.5 23.8 Length Long Long (Groene Calabrese, Premium Crop) Leaf Ratio - 5:1 5:1 Length/Width Leaf Attachment Petiolate Petiolate Wax Presence Intermediate Intermediate Foliage Color (with wax, Dark Green Dark Green if present) Foliage Color (with wax, 139A 136A if present, RHS color chart value) Leaf Shape Narrow Elliptic Narrow Elliptic Leaf Base Blunt Blunt Leaf Apex Blunt Blunt Leaf Margins Very Wavy Very Wavy Leaf Veins Thick Thick Midrib Raised Raised Attitude (leaf angle from Erect Erect ground) (80-100 degrees) Torsion of Leaf Tip Intermediate Intermediate Profile of Upper Side of Concave Planar Leaf Leaf Blade Color Green Green (Claudia, Verflor) Intensity of Color Dark Dark Anthocyanin Coloration Absent Absent (Claudia, Embassy) Undulation of Margin Medium Medium (Citation) Dentation of Margin Medium Medium (Buccaneer) Blistering Medium Medium (Medium Late 145, Skiff) Head Length of Branching at Very Long Short Base (excluding stem) (A Getti di Napoli) Diameter (at widest point, 15.4 14.4 at market maturity, cm) Depth (at market 14.2 10.9 maturity, cm) Weight (market trimmed, 391.4 387.5 at market maturity, grams) Color (at market maturity) Medium Green Dark Green (Idol, Verflor) Intensity of Color Medium Dark Color (at market maturity, 137B N138B RHS color chart value) Anthocyanin Coloration Present Absent (Brigadeer, Shogun, Viola) Intensity of Anthocyanin Weak Coloration (Brigadeer) Shape (at market Circular Transverse Elliptic maturity) (Esquire) Dome Shape (at market Domed Semi Domed maturity) Size (at market maturity) Medium Medium (Dundee, Early Man) Compactness/Firmness (at Short Pedicels/Tight/Firm Short Pedicels/Tight/Firm market maturity) (Captain) Surface Knobbling (at Medium Fine market maturity) (Southern Comet) Texture Coarse Fine (Citation) Bead Size (at market Small Small maturity) Flower Buds (at market Uneven in Size Even In Size maturity) (Cateye) Anthocyanin Coloration Absent Absent of Leaf Axils (at market maturity) Anthocyanin Coloration Absent Absent of Leaf Veins (at market maturity) Anthocyanin Coloration Absent Absent of Leaf Blade (at market maturity) Anthocyanin Coloration Present Present of Entire Plant (at market maturity) Anthocyanin Coloration Absent Absent of Leaf Petiole (at market (Claudia, Embassy) maturity) Color of Head Leaves (at Green Green market maturity) Color of Head Leaves (at 137A 137A market maturity, RHS color chart value) Bracts Absent Absent (Gem, Orion) Secondary Heads (at Axillary along entire main Axillary along entire main market maturity) stem up to main head stem up to main head Prominence of Secondary Weak Weak Heads (at market maturity) Number of Secondary 1.4 1.3 Heads (at market maturity) Flower Color Cream Yellow Intensity of Yellow Color Light Color (RHS color chart 157D 5C value) Stalk Color Green Green Stalk Color (RHS color 137A 137B chart value) Male Sterility Present Present (Chevalier, Montop) *These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.

(19) TABLE-US-00002 TABLE 2 Physiological and Morphological Characteristics of Line BRM 51-1162 CHARACTERISTIC BRM 51-1162 Heritage Species Brassica oleracea var. italica L. Brassica oleracea var. italica L. Region of Adaptation (area Pacific Coast Pacific Coast where best adapted in USA) Maturity Fall Planted (days from 136 129.5 direct seeding to 50% Rep 1 Jan. 8, 2009 rep 2 Jan. 22, 2009 Rep 1 Jan. 8, 2009 rep 2 Jan. 22, 2009 harvest) Rep 1 May 28, 2009 rep 2 Jun. 3, 2009 Rep 1 May 21, 2009 rep 2 May 28, 2009 Fall Planted (days from 86.5 80 transplanting to 50% Rep 1 Feb. 27, 2009 rep 2 Mar. 12, 2009 Rep 1 Feb. 27, 2009 rep 2 Mar. 12, 2009 harvest) Fall Planted (length of 1 1 harvest period in days) Rep 1 May 28, 2009 rep 2 Jun. 3, 2009 Rep 1 May 21, 2009 rep 2 May 28, 2009 Rep 1 May 28, 2009 rep 2 Jun. 3, 2009 Rep 1 May 21, 2009 rep 2 May 28, 2009 Harvest Season (main Spring/Summer Spring/Summer crop at 50% harvest) Time of Harvest Maturity Very Late Medium (50% of plants) (Late Purple Sprouting) Time of Beginning of Medium-Late Medium Flowering (50% of plants with at least 10% flowers) Seedling Cotyledon Color Medium Green Medium Green Cotyledon Color (RHS 132D 132D color chart value) Cotyledon Anthocyanin Weak Weak Hypocotyl Anthocyanin Weak Intermediate Plant Plant Height (at harvest, 52 74.9 cm) Head Height (at harvest, 20.25 38.8 cm) Height (at harvest Short Tall maturity) (Packman, Primor) Number of Stems One One (Ramoso Calabrese, Shogun) Branches Many Medium Habit Compact Intermediate Market Class Both Both Life Cycle Annual Annual Type of Variety Inbred Hybrid Leaf Width Narrow Broad (Arcadia, Brigadeer) Length (including petiole) Long Long (Green Duke, Laser) Number of Lobes Few Many (Early White Sprouting) Attitude (at beginning of Semi-erect Semi Erect head formation) (Arcadia, Asti, Civet, Claudia) Outer Leaves Number of Leaves Per 14.09 14.3 Plant (at harvest) Width (at midpoint of 18.3 18 plant including petiole, cm) Length (at midpoint of 47.8 60.1 plant including petiole, cm) Petiole Length (cm) 18.1 23.8 Leaf Ratio - 4:1 5:1 Length/Width Leaf Attachment Petiolate Petiolate Wax Presence Intermediate Intermediate Foliage Color (with wax, Dark Green Dark Green if present) Foliage Color (with wax, 138A 136A if present, RHS color chart value) Leaf Shape Narrow Elliptic Narrow Elliptic Leaf Base Blunt Blunt Leaf Apex Blunt Blunt Leaf Margins Very Wavy Very Wavy Leaf Veins Thick Thick Midrib Raised Raised Attitude (leaf angle from Erect Erect ground) (80-100 degrees) Torsion of Leaf Tip Intermediate Intermediate Profile of Upper Side of Planar Planar Leaf Petiole Length Long Long (Groene Calabrese, Premium Crop) Leaf Blade Color Green Green (Claudia, Verflor) Intensity of Color Dark Dark Anthocyanin Coloration Present Absent (Buccaneer, Pascal) Undulation of Margin Medium Medium (Citation) Dentation of Margin Medium Medium (Buccaneer) Blistering Medium Medium (Medium Late 145, Skiff) Head Length of Branching at Short Short Base (excluding stem) (Brigadeer, Buccaneer, Emperor) Diameter (at widest point, 13 14.4 at market maturity, cm) Depth (at market 13.2 10.9 maturity, cm) Weight (market trimmed, 355.6 387.5 at market maturity, grams) Color (at market maturity) Blue-green Dark Green (Buccaneer) Intensity of Color Medium Dark Color (at market maturity, 137B N138B RHS color chart value) Anthocyanin Coloration Absent Absent (Early White Sprouting) Shape (at market Circular Transverse Elliptic maturity) (Esquire) Dome Shape (at market Semi-domed Semi Domed maturity) Size (at market maturity) Small Medium (Orbit, Scorpio) Compactness/Firmness (at Short Pedicels/Tight/Firm Short Pedicels/Tight/Firm market maturity) (Captain) Surface Knobbling (at Medium Fine market maturity) (Southern Comet) Texture Fine Fine (Auriga, Bishop, Green Top) Bead Size (at market Small Small maturity) Flower Buds (at market Uneven in Size Even In Size maturity) (Cateye) Anthocyanin Coloration Absent Absent of Leaf Axils (at market maturity) Anthocyanin Coloration Absent Absent of Leaf Veins (at market maturity) Anthocyanin Coloration Absent Absent of Leaf Blade (at market maturity) Anthocyanin Coloration Present Present of Entire Plant (at market maturity) Anthocyanin Coloration Absent Absent of Leaf Petiole (at market (Claudia, Embassy) maturity) Color of Head Leaves (at Green Green market maturity) Color of Head Leaves (at 137A 137A market maturity, RHS color chart value) Bracts Present Absent (Ramoso Calabrese) Secondary Heads (at Completely Absent Axillary along entire main market maturity) (Scorpio, Zeus) stem up to main head Prominence of Secondary Weak Heads (at market maturity) Number of Secondary 0 1.3 Heads (at market maturity) Flower Color Cream Yellow Intensity of Yellow Color Light Color (RHS color chart 155A 5C value) Stalk Color Green Green Stalk Color (RHS color 138A 137B chart value) Male Sterility Absent Present (Marathon) *These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.

(20) TABLE-US-00003 TABLE 3 Physiological and Morphological Characteristics of Line BRL 51-1128 CHARACTERISTIC BRL 51-1128 Heritage Species Brassica oleracea var. italica L. Brassica oleracea var. italica L. Region of Adaptation (area Pacific Coast Pacific Coast where best adapted in USA) Maturity Fall Planted (days from 129.5 129.5 direct seeding to 50% Rep 1 Jan. 8, 2009 rep 2 Jan. 22, 2009 Rep 1 Jan. 8, 2009 rep 2 Jan. 22, 2009 harvest) Rep 1 May 21, 2009 rep 2 May 28, 2009 Rep 1 May 21, 2009 rep 2 May 28, 2009 Fall Planted (days from 80 80 transplanting to 50% Rep 1 Feb. 27, 2009 rep 2 Mar. 12, 2009 Rep 1 Feb. 27, 2009 rep 2 Mar. 12, 2009 harvest) Fall Planted (length of 1 1 harvest period in days) Rep 1 May 21, 2009 rep 2 May 28, 2009 Rep 1 May 21, 2009 rep 2 May 28, 2009 Rep 1 May 21, 2009 rep 2 May 28, 2009 Rep 1 May 21, 2009 rep 2 May 28, 2009 Harvest Season (main Spring/Summer Spring/Summer crop at 50% harvest) Time of Harvest Maturity Early Medium (50% of plants) (Galaxy, Packman, Scorpio) Time of Beginning of Late Medium Flowering (50% of plants (Shogun, Viola) with at least 10% flowers) Seedling Cotyledon Color Medium Green Medium Green Cotyledon Color (RHS 132D 132D color chart value) Cotyledon Anthocyanin Weak Weak Hypocotyl Anthocyanin Strong Intermediate Plant Plant Height (at harvest, 52.6 74.9 cm) Head Height (at harvest, 38.2 38.8 cm) Height (at harvest Short Tall maturity) (Packman, Primor) Number of Stems More Than One One (A Getti di Napoli) Branches Many Medium Habit Compact Intermediate Market Class Both Both Life Cycle Annual Annual Type of Variety Inbred Hybrid Leaf Width Broad Broad (Claudia, Esquire, New Prince) Length (including petiole) Long Long (Green Duke, Laser) Number of Lobes Few Many (Early White Sprouting) Attitude (at beginning of Semi-erect Semi Erect head formation) (Arcadia, Asti, Civet, Claudia) Outer Leaves Number of Leaves Per 27.7 14.3 Plant (at harvest) Width (at midpoint of 11.7 18 plant including petiole, cm) Length (at midpoint of 41 60.1 plant including petiole, cm) Petiole Length (cm) 16.7 23.8 Leaf Ratio - 4:1 5:1 Length/Width Leaf Attachment Petiolate Petiolate Wax Presence Intermediate Intermediate Foliage Color (with wax, Purple-green Dark Green if present) Foliage Color (with wax, 137A 136A if present, RHS color chart value) Leaf Shape Broad Elliptic Narrow Elliptic Leaf Base Blunt Blunt Leaf Apex Blunt Blunt Leaf Margins Slightly Wavy Very Wavy Leaf Veins Thick Thick Midrib Raised Raised Attitude (leaf angle from Erect Erect ground) (80-100 degrees) Torsion of Leaf Tip Intermediate Intermediate Profile of Upper Side of Concave Planar Leaf Petiole Length Long Long (Groene Calabrese, Premium Crop) Leaf Blade Color Green Green (Claudia, Verflor) Intensity of Color Dark Dark Anthocyanin Coloration Present Absent (Buccaneer, Pascal) Undulation of Margin Medium Medium (Citation) Dentation of Margin Medium Medium (Buccaneer) Blistering Medium Medium (Medium Late 145, Skiff) Head Length of Branching at Very Long Short Base (excluding stem) (A Getti di Napoli) Diameter (at widest point, 12.1 14.4 at market maturity, cm) Depth (at market 15.75 10.9 maturity, cm) Weight (market trimmed, 221.8 387.5 at market maturity, grams) Color (at market maturity) Purple/Violet Dark Green (Viola) Intensity of Color Medium Dark Color (at market maturity, 189A N138B RHS color chart value) Anthocyanin Coloration Present Absent (Brigadeer, Shogun, Viola) Intensity of Anthocyanin Strong Coloration Shape (at market Transverse Elliptic Narrow Transverse Elliptic maturity) (Citation, Scorpio, Zeus) Dome Shape (at market Domed Semi Domed maturity) Size (at market maturity) Small Medium (Orbit, Scorpio) Compactness/Firmness (at Short Pedicels/Tight/Firm Short Pedicels/Tight/Firm market maturity) (Captain) Surface Knobbling (at Coarse Fine market maturity) (Perseus, Regilio) Texture Coarse Fine (Citation) Bead Size (at market Small Small maturity) Flower Buds (at market Uneven in Size Even In Size maturity) (Cateye) Anthocyanin Coloration Present Absent of Leaf Axils (at market maturity) Anthocyanin Coloration Absent Absent of Leaf Veins (at market maturity) Anthocyanin Coloration Present Absent of Leaf Blade (at market maturity) Anthocyanin Coloration Present Present of Entire Plant (at market maturity) Anthocyanin Coloration Absent Absent of Leaf Petiole (at market (Claudia, Embassy) maturity) Color of Head Leaves (at Green Green market maturity) Color of Head Leaves (at 139A 137A market maturity, RHS color chart value) Bracts Present Absent (Ramoso Calabrese) Secondary Heads (at Basal Axillary along entire main market maturity) stem up to main head Prominence of Secondary Strong Weak Heads (at market (Marathon, Tribute) maturity) Number of Secondary 1.5 1.3 Heads (at market maturity) Flower Color Yellow Yellow (Brigadeer, Orion) Intensity of Yellow Color Medium Light (Capitol, Corvet) Color (RHS color chart 3D 5C value) Stalk Color Green Green Stalk Color (RHS color 137B 137B chart value) Male Sterility Present Present (Chevalier, Montop) *These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.

C. BREEDING BROCCOLI PLANTS

(21) One aspect of the current invention concerns methods for producing seed of broccoli hybrid PS05151639 involving crossing broccoli lines BRM 51-1162 and BRL 51-1128. Alternatively, in other embodiments of the invention, hybrid PS05151639, line BRM 51-1162, or line BRL 51-1128 may be crossed with itself or with any second plant. Such methods can be used for propagation of hybrid PS05151639 and/or the broccoli lines BRM 51-1162 and BRL 51-1128, or can be used to produce plants that are derived from hybrid PS05151639 and/or the broccoli lines BRM 51-1162 and BRL 51-1128. Plants derived from hybrid PS05151639 and/or the broccoli lines BRM 51-1162 and BRL 51-1128 may be used, in certain embodiments, for the development of new broccoli varieties.

(22) The development of new varieties using one or more starting varieties is well known in the art. In accordance with the invention, novel varieties may be created by crossing hybrid PS05151639 followed by multiple generations of breeding according to such well known methods. New varieties may be created by crossing with any second plant. In selecting such a second plant to cross for the purpose of developing novel lines, it may be desired to choose those plants which either themselves exhibit one or more selected desirable characteristics or which exhibit the desired characteristic(s) when in hybrid combination. Once initial crosses have been made, inbreeding and selection take place to produce new varieties. For development of a uniform line, often five or more generations of selfing and selection are involved.

(23) Uniform lines of new varieties may also be developed by way of double-haploids. This technique allows the creation of true breeding lines without the need for multiple generations of selfing and selection. In this manner true breeding lines can be produced in as little as one generation. Haploid embryos may be produced from microspores, pollen, anther cultures, or ovary cultures. The haploid embryos may then be doubled autonomously, or by chemical treatments (e.g. colchicine treatment). Alternatively, haploid embryos may be grown into haploid plants and treated to induce chromosome doubling. In either case, fertile homozygous plants are obtained. In accordance with the invention, any of such techniques may be used in connection with a plant of the invention and progeny thereof to achieve a homozygous line.

(24) Backcrossing can also be used to improve an inbred plant. Backcrossing transfers a specific desirable trait from one inbred or non-inbred source to an inbred that lacks that trait. This can be accomplished, for example, by first crossing a superior inbred (A) (recurrent parent) to a donor inbred (non-recurrent parent), which carries the appropriate locus or loci for the trait in question. The progeny of this cross are then mated back to the superior recurrent parent (A) followed by selection in the resultant progeny for the desired trait to be transferred from the non-recurrent parent. After five or more backcross generations with selection for the desired trait, the progeny have the characteristic being transferred, but are like the superior parent for most or almost all other loci. The last backcross generation would be selfed to give pure breeding progeny for the trait being transferred.

(25) The plant of the present invention are particularly well suited for the development of new lines based on the elite nature of the genetic background of the plants. In selecting a second plant to cross with PS05151639 and/or broccoli lines BRM 51-1162 and BRL 51-1128 for the purpose of developing novel broccoli lines, it will typically be preferred to choose those plants which either themselves exhibit one or more selected desirable characteristics or which exhibit the desired characteristic(s) when in hybrid combination. Examples of desirable traits may include, in specific embodiments, high seed yield, high seed germination, seedling vigor, high yield, disease tolerance or resistance, and adaptability for soil and climate conditions. Consumer-driven traits, such as a head shape, nutritional value, and taste are other traits that may be incorporated into new lines of broccoli plants developed by this invention.

D. PERFORMANCE CHARACTERISTICS

(26) As described above, hybrid PS05151639 exhibits desirable agronomic traits. The performance characteristics of PS05151639 were the subject of an objective analysis of the performance traits relative to other varieties. The results of the analysis are presented below.

(27) TABLE-US-00004 TABLE 4 Performance Characteristics For PS05151639 and Selected Varieties* Stem Traits Variety Plant Traits Smoothness Cleaning Lateral (average Size Type Uniformity Vigor (1 = very Side Shoots Size 1 = very smooth, ability Heads over eight (1 = tall, (1 = erect, (1 = excellent, strong, 9 = (1 = absent, (1 = thick, 9 = very (1 = very easy, (1 = absent. trials) 9 = short) 9 = open) 9 = poor) very weak) 9 = very weak) 9 = thin) irregular) 9 = difficult) 9 = alot) PS05151639 3.6 2.3 3.79 3.3 1.5 5.8 1.92 1.5 1 Average Marathon 4.8 3.5 3 3.3 2.125 6 1.75 1.56 1 Average Heritage 3.5 2.3 3 3.3 2.25 4.6 4.42 4.06 1 Average Variety Head Traits (average Floret Traits Size (cm) (1 = extra over eight Size (1 = tall, Contrast Color Pedicel Size (1 = short, Type (1 = crown, large [>7], 7 = small Color (1 = dark green, trials) 9 = short) (1 = absent, 9 = >26%) 9 = large) 9 = spear) [<5]) 9 = purple green) PS05151639 4 5 1 1 5 3.59 Average Marathon 4 5 1 1 5 4.34 Average Heritage 6 5 1 1 5 1.58 average Variety Head Traits (average Smoothness (1 = very Weight (1 = very heavy over eight Shape (1 = high dome, Bead Size (1 = very Firmness (1 = very firm, smooth, 9 = very [<400 gr], 9 = light trials) 9 = flat) fine, 9 = variable) 9 = soft) irregular) [<200 gr]) PS05151639 2.4 4 5.2 5.2 5.25 Average Marathon 3.9 4.05 5.1 4.3 6.75 Average Heritage 3.1 4.125 3.3 3.3 5 average Defects Harvest Variety Hollow Stem Stem Holding Overall (average (1 = 100% steam Oxidation Bracting Cat eye Brown bead Open bead Diseases Ability (field) (1 = over eight solid, 9 = 76-100% (1 = absent, (1 = absent, (1 = absent, (1 = absent, (1 = absent, (1 = absent, (1 = good, advance, trials) hollow) 9 = present) 9 = severe) 9 = severe) 9 = severe) 9 = present) 9 = severe) 9 = bad) 9 = drop) PS05151639 1 1 1 2.5 1.06 1 1 1.625 4.8 Average Marathon 1 1 1 2.6 1 1 1 1.625 4.9 Average Heritage 1.125 1 1 2.5 1 1 1 1 3.5 Average *Based on scales as indicated

(28) MSB levels were quantitated using standard methods. The data was analyzed using a combined analysis across locations and plantings, considering variety as a fixed effect, and location, planting, reps within (locationplanting), locationplanting, and their interactions with variety as random effects. Results for MSB are presented in Table 5. Hybrid PS05151639's LS-mean was 428, vs comparison varieties Heritage LSM of 180 and Marathon's of 123. PS05151639 contained 2.4 more MSB than Heritage and 3.5 more than Marathon.

(29) TABLE-US-00005 TABLE 5 MSB Levels For PS05151639 and Selected Varieties Variety MSB LSM Std. Error Difference X-Difference PS05151639 428 46.2 Heritage 180 46.2 249 2.4X Marathon 123 46.2 305 3.5X

(30) MSP levels were quantitated using standard methods. The data was analyzed using a combined analysis across locations and plantings, considering variety as fixed effect, and location, planting, reps within (locationplanting), locationplanting, and their interactions with variety as random effects. Results for MSP are presented in Table 6. Hybrid PS05151639's LS-mean was 62, vs comparison varieties Heritage LSM of 8 and Marathon's LSM of 7. PS05151639's MSB was 7.8 that of Heritage and 8.5 that of Marathon.

(31) TABLE-US-00006 TABLE 6 Performance Characteristics For PS05151639 and Selected Varieties Variety MSP LSM Std. Error Difference X-Difference PS05151639 62 8.2 Heritage 8 8.2 54 7.8X Marathon 7 8.2 55 8.5X

(32) MSB and MSP levels were quantitated in the lines BRL 51-1128 and BRM 51-1162 using standard methods. Results of this analysis are presented in Table 7.

(33) TABLE-US-00007 TABLE 7 Performance Characteristics For BRL 51-1128 and BRM 51-1162 Glucoraphanin (MSB) Glucoiberin (MSP) micromoles/gram micromoles/gram Variety fresh weight fresh weight BRL 51-1128 CMS 1.026 0.000* BRL 51-1128 CMS 1.256 0.000* BRL 51-1128 CMS 1.420 0.168 BRM 51-1162 0.785 0.798 BRM 51-1162 SC 1.092 0.972 BRM 51-1162 SC 1.237 1.089 BRM 51-1162 SC 1.598 1.402 BRM 51-1162 SC 1.552 1.531 BRM 51-1162 SC 1.933 1.609 BRM 51-1162 SC 1.230 1.431 BRM 51-1162 SC 1.480 1.761 BRM 51-1162 SC 1.440 1.912 BRM 51-1162 SC 1.606 1.806 BRM 51-1162 SC 1.873 2.444 BRM 51-1162 SC 2.237 2.443 BRM 51-1162 SC 2.145 2.032 *no measurable glucoiberin (MSP)

E. FURTHER EMBODIMENTS OF THE INVENTION

(34) In certain aspects of the invention, plants described herein are provided modified to include at least a first desired heritable trait. Such plants may, in one embodiment, be developed by a plant breeding technique called backcrossing, wherein essentially all of the desired morphological and physiological characteristics of a variety are recovered in addition to a genetic locus transferred into the plant via the backcrossing technique. The term single locus converted plant as used herein refers to those broccoli plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the desired morphological and physiological characteristics of a variety are recovered in addition to the single locus transferred into the variety via the backcrossing technique. By essentially all of the desired morphological and physiological characteristics, it is meant that the characteristics of a plant are recovered that are otherwise present when compared in the same environment, other than an occasional variant trait that might arise during backcrossing or direct introduction of a transgene.

(35) Backcrossing methods can be used with the present invention to improve or introduce a characteristic into the present variety. The parental broccoli plant which contributes the locus for the desired characteristic is termed the nonrecurrent or donor parent. This terminology refers to the fact that the nonrecurrent parent is used one time in the backcross protocol and therefore does not recur. The parental broccoli plant to which the locus or loci from the nonrecurrent parent are transferred is known as the recurrent parent as it is used for several rounds in the backcrossing protocol.

(36) In a typical backcross protocol, the original variety of interest (recurrent parent) is crossed to a second variety (nonrecurrent parent) that carries the single locus of interest to be transferred. The resulting progeny from this cross are then crossed again to the recurrent parent and the process is repeated until a broccoli plant is obtained wherein essentially all of the desired morphological and physiological characteristics of the recurrent parent are recovered in the converted plant, in addition to the single transferred locus from the nonrecurrent parent.

(37) The selection of a suitable recurrent parent is an important step for a successful backcrossing procedure. The goal of a backcross protocol is to alter or substitute a single trait or characteristic in the original variety. To accomplish this, a single locus of the recurrent variety is modified or substituted with the desired locus from the nonrecurrent parent, while retaining essentially all of the rest of the desired genetic, and therefore the desired physiological and morphological constitution of the original variety. The choice of the particular nonrecurrent parent will depend on the purpose of the backcross; one of the major purposes is to add some commercially desirable trait to the plant. The exact backcrossing protocol will depend on the characteristic or trait being altered and the genetic distance between the recurrent and nonrecurrent parents. Although backcrossing methods are simplified when the characteristic being transferred is a dominant allele, a recessive allele, or an additive allele (between recessive and dominant), may also be transferred. In this instance it may be necessary to introduce a test of the progeny to determine if the desired characteristic has been successfully transferred.

(38) In one embodiment, progeny broccoli plants of a backcross in which PS05151639 is the recurrent parent comprise (i) the desired trait from the non-recurrent parent and (ii) all of the physiological and morphological characteristics of broccoli hybrid PS05151639 as determined at the 5% significance level when grown in the same environmental conditions.

(39) Broccoli varieties can also be developed from more than two parents. The technique, known as modified backcrossing, uses different recurrent parents during the backcrossing. Modified backcrossing may be used to replace the original recurrent parent with a variety having certain more desirable characteristics or multiple parents may be used to obtain different desirable characteristics from each.

(40) Many single locus traits have been identified that are not regularly selected for in the development of a new inbred but that can be improved by backcrossing techniques. Single locus traits may or may not be transgenic; examples of these traits include, but are not limited to, herbicide resistance, resistance to bacterial, fungal, or viral disease, insect resistance, modified fatty acid or carbohydrate metabolism, and altered nutritional quality. These comprise genes generally inherited through the nucleus.

(41) Direct selection may be applied where the single locus acts as a dominant trait. For this selection process, the progeny of the initial cross are assayed for viral resistance and/or the presence of the corresponding gene prior to the backcrossing. Selection eliminates any plants that do not have the desired gene and resistance trait, and only those plants that have the trait are used in the subsequent backcross. This process is then repeated for all additional backcross generations.

(42) Selection of broccoli plants for breeding is not necessarily dependent on the phenotype of a plant and instead can be based on genetic investigations. For example, one can utilize a suitable genetic marker which is closely genetically linked to a trait of interest. One of these markers can be used to identify the presence or absence of a trait in the offspring of a particular cross, and can be used in selection of progeny for continued breeding. This technique is commonly referred to as marker assisted selection. Any other type of genetic marker or other assay which is able to identify the relative presence or absence of a trait of interest in a plant can also be useful for breeding purposes. Procedures for marker assisted selection are well known in the art. Such methods will be of particular utility in the case of recessive traits and variable phenotypes, or where conventional assays may be more expensive, time consuming or otherwise disadvantageous. Types of genetic markers which could be used in accordance with the invention include, but are not necessarily limited to, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., 1998).

F. PLANTS DERIVED BY GENETIC ENGINEERING

(43) Many useful traits that can be introduced by backcrossing, as well as directly into a plant, are those which are introduced by genetic transformation techniques. Genetic transformation may therefore be used to insert a selected transgene into a plant of the invention or may, alternatively, be used for the preparation of transgenes which can be introduced by backcrossing. Methods for the transformation of plants that are well known to those of skill in the art and applicable to many crop species include, but are not limited to, electroporation, microprojectile bombardment, Agrobacterium-mediated transformation and direct DNA uptake by protoplasts.

(44) To effect transformation by electroporation, one may employ either friable tissues, such as a suspension culture of cells or embryogenic callus or alternatively one may transform immature embryos or other organized tissue directly. In this technique, one would partially degrade the cell walls of the chosen cells by exposing them to pectin-degrading enzymes (pectolyases) or mechanically wound tissues in a controlled manner.

(45) An efficient method for delivering transforming DNA segments to plant cells is microprojectile bombardment. In this method, particles are coated with nucleic acids and delivered into cells by a propelling force. Exemplary particles include those comprised of tungsten, platinum, and preferably, gold. For the bombardment, cells in suspension are concentrated on filters or solid culture medium. Alternatively, immature embryos or other target cells may be arranged on solid culture medium. The cells to be bombarded are positioned at an appropriate distance below the macroprojectile stopping plate.

(46) An illustrative embodiment of a method for delivering DNA into plant cells by acceleration is the Biolistics Particle Delivery System, which can be used to propel particles coated with DNA or cells through a screen, such as a stainless steel or Nytex screen, onto a surface covered with target cells. The screen disperses the particles so that they are not delivered to the recipient cells in large aggregates. It is believed that a screen intervening between the projectile apparatus and the cells to be bombarded reduces the size of projectiles aggregate and may contribute to a higher frequency of transformation by reducing the damage inflicted on the recipient cells by projectiles that are too large. Microprojectile bombardment techniques are widely applicable, and may be used to transform virtually any plant species.

(47) Agrobacterium-mediated transfer is another widely applicable system for introducing gene loci into plant cells. An advantage of the technique is that DNA can be introduced into whole plant tissues, thereby bypassing the need for regeneration of an intact plant from a protoplast. Modern Agrobacterium transformation vectors are capable of replication in E. coli as well as Agrobacterium, allowing for convenient manipulations (Klee et al., 1985). Moreover, recent technological advances in vectors for Agrobacterium-mediated gene transfer have improved the arrangement of genes and restriction sites in the vectors to facilitate the construction of vectors capable of expressing various polypeptide coding genes. The vectors described have convenient multi-linker regions flanked by a promoter and a polyadenylation site for direct expression of inserted polypeptide coding genes. Additionally, Agrobacterium containing both armed and disarmed Ti genes can be used for transformation.

(48) In those plant strains where Agrobacterium-mediated transformation is efficient, it is the method of choice because of the facile and defined nature of the gene locus transfer. The use of Agrobacterium-mediated plant integrating vectors to introduce DNA into plant cells is well known in the art (Fraley et al., 1985; U.S. Pat. No. 5,563,055).

(49) Transformation of plant protoplasts also can be achieved using methods based on calcium phosphate precipitation, polyethylene glycol treatment, electroporation, and combinations of these treatments (see, e.g., Potrykus et al., 1985; Omirulleh et al., 1993; Fromm et al., 1986; Uchimiya et al., 1986; Marcotte et al., 1988). Transformation of plants and expression of foreign genetic elements is exemplified in Choi et al. (1994), and Ellul et al. (2003).

(50) A number of promoters have utility for plant gene expression for any gene of interest including but not limited to selectable markers, scoreable markers, genes for pest tolerance, disease resistance, nutritional enhancements and any other gene of agronomic interest. Examples of constitutive promoters useful for broccoli plant gene expression include, but are not limited to, the cauliflower mosaic virus (CaMV) P-35S promoter, which confers constitutive, high-level expression in most plant tissues (see, e.g., Odel et al., 1985), including monocots (see, e.g., Dekeyser et al., 1990; Terada and Shimamoto, 1990); a tandemly duplicated version of the CaMV 35S promoter, the enhanced 35S promoter (P-e35S) the nopaline synthase promoter (An et al., 1988), the octopine synthase promoter (Fromm et al., 1989); and the figwort mosaic virus (P-FMV) promoter as described in U.S. Pat. No. 5,378,619 and an enhanced version of the FMV promoter (P-eFMV) where the promoter sequence of P-FMV is duplicated in tandem, the cauliflower mosaic virus 19S promoter, a sugarcane bacilliform virus promoter, a commelina yellow mottle virus promoter, and other plant DNA virus promoters known to express in plant cells.

(51) A variety of plant gene promoters that are regulated in response to environmental, hormonal, chemical, and/or developmental signals can be used for expression of an operably linked gene in plant cells, including promoters regulated by (1) heat (Callis et al., 1988), (2) light (e.g., pea rbcS-3A promoter, Kuhlemeier et al., 1989; maize rbcS promoter, Schaffner and Sheen, 1991; or chlorophyll a/b-binding protein promoter, Simpson et al., 1985), (3) hormones, such as abscisic acid (Marcotte et al., 1989), (4) wounding (e.g., wunl, Siebertz et al., 1989); or (5) chemicals such as methyl jasmonate, salicylic acid, or Safener. It may also be advantageous to employ organ-specific promoters (e.g., Roshal et al., 1987; Schernthaner et al., 1988; Bustos et al., 1989).

(52) Exemplary nucleic acids which may be introduced to the broccoli plants of this invention include, for example, DNA sequences or genes from another species, or even genes or sequences which originate with or are present in the same species, but are incorporated into recipient cells by genetic engineering methods rather than classical reproduction or breeding techniques. However, the term exogenous is also intended to refer to genes that are not normally present in the cell being transformed, or perhaps simply not present in the form, structure, etc., as found in the transforming DNA segment or gene, or genes which are normally present and that one desires to express in a manner that differs from the natural expression pattern, e.g., to over-express. Thus, the term exogenous gene or DNA is intended to refer to any gene or DNA segment that is introduced into a recipient cell, regardless of whether a similar gene may already be present in such a cell. The type of DNA included in the exogenous DNA can include DNA which is already present in the plant cell, DNA from another plant, DNA from a different organism, or a DNA generated externally, such as a DNA sequence containing an antisense message of a gene, or a DNA sequence encoding a synthetic or modified version of a gene.

(53) Many hundreds if not thousands of different genes are known and could potentially be introduced into a broccoli plant according to the invention. Non-limiting examples of particular genes and corresponding phenotypes one may choose to introduce into a broccoli plant include one or more genes for insect tolerance, such as a Bacillus thuringiensis (B.t.) gene, pest tolerance such as genes for fungal disease control, herbicide tolerance such as genes conferring glyphosate tolerance, and genes for quality improvements such as yield, nutritional enhancements, environmental or stress tolerances, or any desirable changes in plant physiology, growth, development, morphology or plant product(s). For example, structural genes would include any gene that confers insect tolerance including but not limited to a Bacillus insect control protein gene as described in WO 99/31248, herein incorporated by reference in its entirety, U.S. Pat. No. 5,689,052, herein incorporated by reference in its entirety, U.S. Pat. Nos. 5,500,365 and 5,880,275, herein incorporated by reference it their entirety. In another embodiment, the structural gene can confer tolerance to the herbicide glyphosate as conferred by genes including, but not limited to Agrobacterium strain CP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat. No. 5,633,435, herein incorporated by reference in its entirety, or glyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No. 5,463,175, herein incorporated by reference in its entirety.

(54) Alternatively, the DNA coding sequences can affect these phenotypes by encoding a non-translatable RNA molecule that causes the targeted inhibition of expression of an endogenous gene, for example via antisense- or cosuppression-mediated mechanisms (see, for example, Bird et al., 1991). The RNA could also be a catalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desired endogenous mRNA product (see for example, Gibson and Shillito, 1997). Thus, any gene which produces a protein or mRNA which expresses a phenotype or morphology change of interest is useful for the practice of the present invention.

G. DEFINITIONS

(55) In the description and tables herein, a number of terms are used. In order to provide a clear and consistent understanding of the specification and claims, the following definitions are provided:

(56) Allele: Any of one or more alternative forms of a gene locus, all of which alleles relate to one trait or characteristic. In a diploid cell or organism, the two alleles of a given gene occupy corresponding loci on a pair of homologous chromosomes.

(57) Backcrossing: A process in which a breeder repeatedly crosses hybrid progeny, for example a first generation hybrid (F.sub.1), back to one of the parents of the hybrid progeny. Backcrossing can be used to introduce one or more single locus conversions from one genetic background into another.

(58) Crossing: The mating of two parent plants.

(59) Cross-pollination: Fertilization by the union of two gametes from different plants.

(60) Diploid: A cell or organism having two sets of chromosomes.

(61) Emasculate: The removal of plant male sex organs or the inactivation of the organs with a cytoplasmic or nuclear genetic factor or a chemical agent conferring male sterility.

(62) Enzymes: Molecules which can act as catalysts in biological reactions.

(63) F.sub.1 Hybrid: The first generation progeny of the cross of two nonisogenic plants.

(64) Genotype: The genetic constitution of a cell or organism.

(65) Haploid: A cell or organism having one set of the two sets of chromosomes in a diploid.

(66) Linkage: A phenomenon wherein alleles on the same chromosome tend to segregate together more often than expected by chance if their transmission was independent.

(67) Marker: A readily detectable phenotype, preferably inherited in codominant fashion (both alleles at a locus in a diploid heterozygote are readily detectable), with no environmental variance component, i.e., heritability of 1.

(68) Phenotype: The detectable characteristics of a cell or organism, which characteristics are the manifestation of gene expression.

(69) Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer to genetic loci that control to some degree numerically representable traits that are usually continuously distributed.

(70) Resistance: As used herein, the terms resistance and tolerance are used interchangeably to describe plants that show no symptoms to a specified biotic pest, pathogen, abiotic influence or environmental condition. These terms are also used to describe plants showing some symptoms but that are still able to produce marketable product with an acceptable yield. Some plants that are referred to as resistant or tolerant are only so in the sense that they may still produce a crop, even though the plants are stunted and the yield is reduced.

(71) Regeneration: The development of a plant from tissue culture.

(72) Royal Horticultural Society (RHS) color chart value: The RHS color chart is a standardized reference which allows accurate identification of any color. A color's designation on the chart describes its hue, brightness and saturation. A color is precisely named by the RHS color chart by identifying the group name, sheet number and letter, e.g., Yellow-Orange Group 19A or Red Group 41B.

(73) Self-pollination: The transfer of pollen from the anther to the stigma of the same plant.

(74) Single Locus Converted (Conversion) Plant: Plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the desired morphological and physiological characteristics of a broccoli variety are recovered in addition to the characteristics of the single locus transferred into the variety via the backcrossing technique and/or by genetic transformation.

(75) Substantially Equivalent: A characteristic that, when compared, does not show a statistically significant difference (e.g., p=0.05) from the mean.

(76) Tissue Culture: A composition comprising isolated cells of the same or a different type or a collection of such cells organized into parts of a plant.

(77) Transgene: A genetic locus comprising a sequence which has been introduced into the genome of a broccoli plant by transformation.

H. DEPOSIT INFORMATION

(78) A deposit of broccoli hybrid PS05151639 and lines BRM 51-1162 and BRL 51-1128, disclosed above and recited in the claims, has been made with the American Type Culture Collection (ATCC), 10801 University Blvd., Manassas, Va. 20110-2209. The date of the deposits made was Dec. 23, 2008. The accession numbers for those deposited seeds of broccoli hybrid PS05151639 and inbred parent lines BRM 51-1162 and BRL 51-1128 are ATCC Accession Number PTA-9676, ATCC Accession Number PTA-9675, and ATCC Accession Number PTA-9674, respectively. Upon issuance of a patent, all restrictions upon the deposits will be removed, and the deposits are intended to meet all of the requirements of 37 C.F.R. 1.801-1.809. The deposits will be maintained in the depository for a period of 30 years, or 5 years after the last request, or for the effective life of the patent, whichever is longer, and will be replaced if necessary during that period.

(79) Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity and understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the invention, as limited only by the scope of the appended claims.

(80) All references cited herein are hereby expressly incorporated herein by reference.

REFERENCES

(81) The following references, to the extent that they provide exemplary procedural or other details supplementary to those set forth herein, are specifically incorporated herein by reference: U.S. Pat. No. 5,378,619 U.S. Pat. No. 5,463,175 U.S. Pat. No. 5,500,365 U.S. Pat. No. 5,563,055 U.S. Pat. No. 5,633,435 U.S. Pat. No. 5,689,052 U.S. Pat. No. 5,880,275 An et al., Plant Physiol., 88:547, 1988. Bird et al., Biotech. Gen. Engin. Rev., 9:207, 1991. Bustos et al., Plant Cell, 1:839, 1989. Callis et al., Plant Physiol., 88:965, 1988. Choi et al., Plant Cell Rep., 13: 344-348, 1994. Dekeyser et al., Plant Cell, 2:591, 1990. Ellul et al., Theor. Appl. Genet., 107:462-469, 2003. EP 534 858 Fraley et al., Bio/Technology, 3:629-635, 1985. Fromm et al., Nature, 312:791-793, 1986. Fromm et al., Plant Cell, 1:977, 1989. Gibson and Shillito, Mol. Biotech., 7:125, 1997 Klee et al., Bio-Technology, 3(7):637-642, 1985. Kuhlemeier et al., Plant Cell, 1:471, 1989. Marcotte et al., Nature, 335:454, 1988. Marcotte et al., Plant Cell, 1:969, 1989. Odel et al., Nature, 313:810, 1985. Omirulleh et al., Plant Mol. Biol., 21(3):415-428, 1993. Potrykus et al., Mol. Gen. Genet., 199:183-188, 1985. Roshal et al., EMBO J., 6:1155, 1987. Schaffner and Sheen, Plant Cell, 3:997, 1991. Schernthaner et al., EMBO J., 7:1249, 1988. Siebertz et al., Plant Cell, 1:961, 1989. Simpson et al., EMBO J., 4:2723, 1985. Terada and Shimamoto, Mol. Gen. Genet., 220:389, 1990. Uchimiya et al., Mol. Gen. Genet., 204:204, 1986. Wang et al., Science, 280:1077-1082, 1998. Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990. WO 99/31248