# Patent

## Title
Tomato hybrid PX 02481245 and parents thereof

## Abstract

The invention provides seed and plants of tomato hybrid PX 02481245 and the parent lines thereof. The invention thus relates to the plants, seeds and tissue cultures of tomato hybrid PX 02481245 and the parent lines thereof, and to methods for producing a tomato plant produced by crossing such plants with themselves or with another tomato 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, including the fruit and gametes of such plants.

## Background
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a divisional of U.S. Ser. No. 13/715,796, filed Dec. 14, 2012, now U.S. Pat. No. 8,952,227, issued on Feb. 10, 2015, which claims the priority of U.S. Provisional Appl. Ser. No. 61/576,697, filed Dec. 16, 2011, the entire disclosures of which are incorporated herein by reference in their entireties.


FIELD OF THE INVENTION
The present invention relates to the field of plant breeding and, more specifically, to the development of tomato hybrid PX 02481245 and the inbred tomato lines PSQ 24-2082 and PSQ-9Z08005.
BACKGROUND OF THE INVENTION
The goal of vegetable breeding is to combine various desirable traits in a single variety/hybrid. Such desirable traits may include any trait deemed beneficial by a grower and/or consumer, including greater yield, resistance to insects or disease, tolerance to environmental stress, and nutritional value.
Breeding techniques take advantage of a plant's method of pollination. There are two general methods of pollination: a plant self-pollinates if pollen from one flower is transferred to the same or another flower of the same plant or plant variety. A plant cross-pollinates if pollen comes to it from a flower of a different plant variety.
Plants that have been self-pollinated and selected for type over many generations become homozygous at almost all gene loci and produce a uniform population of true breeding progeny, a homozygous plant. A cross between two such homozygous plants of different genotypes produces a uniform population of hybrid plants that are heterozygous for many gene loci. Conversely, a cross of two plants each heterozygous at a number of loci produces a population of hybrid plants that differ genetically and are not uniform. The resulting non-uniformity makes performance unpredictable.
The development of uniform varieties requires the development of homozygous inbred plants, the crossing of these inbred plants, and the evaluation of the crosses. Pedigree breeding and recurrent selection are examples of breeding methods that have been used to develop inbred plants from breeding populations. Those breeding methods combine the genetic backgrounds from two or more plants or various other broad-based sources into breeding pools from which new lines and hybrids derived therefrom are developed by selfing and selection of desired phenotypes. The new lines and hybrids are evaluated to determine which of those have commercial potential.
SUMMARY OF THE INVENTION
In one aspect, the present invention provides a tomato plant of the hybrid designated PX 02481245, the tomato line PSQ 24-2082 or tomato line PSQ-9Z08005. Also provided are tomato plants having all the physiological and morphological characteristics of such a plant. Parts of these tomato plants are also provided, for example, including pollen, an ovule, scion, a rootstock, a fruit, and a cell of the plant.
In another aspect of the invention, a plant of tomato hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005 comprising an added heritable trait is provided. The heritable trait may comprise a genetic locus that is, for example, a dominant or recessive allele. In one embodiment of the invention, a plant of tomato hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005 is defined as comprising a single locus conversion. In specific embodiments of the invention, an added genetic locus confers one or more traits such as, for example, herbicide tolerance, insect resistance, disease resistance, and modified carbohydrate metabolism. In further embodiments, the trait may be conferred by a naturally occurring gene introduced into the genome of a line by backcrossing, a natural or induced mutation, or a transgene introduced through genetic transformation techniques into the plant or a progenitor of any previous generation thereof. When introduced through transformation, a genetic locus may comprise one or more genes integrated at a single chromosomal location.
The invention also concerns the seed of tomato hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005. The tomato seed of the invention may be provided as an essentially homogeneous population of tomato seed of tomato hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005. Essentially homogeneous populations of seed are generally free from substantial numbers of other seed. Therefore, in some embodiments, seed of hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005 may be defined as forming at least about 97% of the total seed, including at least about 98%, 99% or more of the seed. The seed population may be separately grown to provide an essentially homogeneous population of tomato plants designated PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005.
In yet another aspect of the invention, a tissue culture of regenerable cells of a tomato plant of hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005 is provided. The tissue culture will preferably be capable of regenerating tomato plants capable of expressing all of the physiological and morphological characteristics of the starting plant, and of regenerating plants having substantially the same genotype as the starting plant. Examples of some of the physiological and morphological characteristics of the hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005 include those traits set forth in the tables herein. The regenerable cells in such tissue cultures may be derived, for example, from embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistils, flowers, seed and stalks. Still further, the present invention provides tomato plants regenerated from a tissue culture of the invention, the plants having all the physiological and morphological characteristics of hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005.
In still yet another aspect of the invention, processes are provided for producing tomato seeds, plants and fruit, which processes generally comprise crossing a first parent tomato plant with a second parent tomato plant, wherein at least one of the first or second parent tomato plants is a plant of tomato line PSQ 24-2082 or tomato line PSQ-9Z08005. These processes may be further exemplified as processes for preparing hybrid tomato seed or plants, wherein a first tomato plant is crossed with a second tomato plant of a different, distinct genotype to provide a hybrid that has, as one of its parents, a plant of tomato line PSQ 24-2082 or tomato line PSQ-9Z08005. In these processes, crossing will result in the production of seed. The seed production occurs regardless of whether the seed is collected or not.
In one embodiment of the invention, the first step in “crossing” comprises planting seeds of a first and second parent tomato plant, often in proximity so that pollination will occur for example, mediated by insect vectors. Alternatively, pollen can be transferred manually. Where the plant is self-pollinated, pollination may occur without the need for direct human intervention other than plant cultivation.
A second step may comprise cultivating or growing the seeds of first and second parent tomato plants into plants that bear flowers. A third step may comprise preventing self-pollination of the plants, such as by emasculating the flowers (i.e., killing or removing the pollen).
A fourth step for a hybrid cross may comprise cross-pollination between the first and second parent tomato plants. Yet another step comprises harvesting the seeds from at least one of the parent tomato plants. The harvested seed can be grown to produce a tomato plant or hybrid tomato plant.
The present invention also provides the tomato seeds and plants produced by a process that comprises crossing a first parent tomato plant with a second parent tomato plant, wherein at least one of the first or second parent tomato plants is a plant of tomato hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005. In one embodiment of the invention, tomato seed and plants produced by the process are first generation (F1) hybrid tomato seed and plants produced by crossing a plant in accordance with the invention with another, distinct plant. The present invention further contemplates plant parts of such an F1 hybrid tomato plant, and methods of use thereof. Therefore, certain exemplary embodiments of the invention provide an F1 hybrid tomato plant and seed thereof.
In still yet another aspect, the present invention provides a method of producing a plant derived from hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005, the method comprising the steps of: (a) preparing a progeny plant derived from hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005, wherein said preparing comprises crossing a plant of the hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005 with a second plant; and (b) crossing the progeny plant with itself or a second plant to produce a seed of a progeny plant of a subsequent generation. In further embodiments, the method may additionally comprise: (c) growing a progeny plant of a subsequent generation from said seed of a progeny plant of a subsequent generation and crossing the progeny plant of a subsequent generation with itself or a second plant; and repeating the steps for an additional 3-10 generations to produce a plant derived from hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005. The plant derived from hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005 may be an inbred line, and the aforementioned repeated crossing steps may be defined as comprising sufficient inbreeding to produce the inbred line. In the method, it may be desirable to select particular plants resulting from step (c) for continued crossing according to steps (b) and (c). By selecting plants having one or more desirable traits, a plant derived from hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005 is obtained which possesses some of the desirable traits of the line/hybrid as well as potentially other selected traits.
In certain embodiments, the present invention provides a method of producing food or feed comprising: (a) obtaining a plant of tomato hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005, wherein the plant has been cultivated to maturity, and (b) collecting at least one tomato from the plant.
In still yet another aspect of the invention, the genetic complement of tomato hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005 is provided. The phrase “genetic complement” is used to refer to the aggregate of nucleotide sequences, the expression of which sequences defines the phenotype of, in the present case, a tomato plant, or a cell or tissue of that plant. A genetic complement thus represents the genetic makeup of a cell, tissue or plant, and a hybrid genetic complement represents the genetic make up of a hybrid cell, tissue or plant. The invention thus provides tomato plant cells that have a genetic complement in accordance with the tomato plant cells disclosed herein, and seeds and plants containing such cells.
Plant genetic complements may be assessed by genetic marker profiles, and by the expression of phenotypic traits that are characteristic of the expression of the genetic complement, e.g., isozyme typing profiles. It is understood that hybrid PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005 could be identified by any of the many well known techniques such as, for example, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., Nucleic Acids Res., 1 8:6531-6535, 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., Science, 280:1077-1082, 1998).
In still yet another aspect, the present invention provides hybrid genetic complements, as represented by tomato plant cells, tissues, plants, and seeds, formed by the combination of a haploid genetic complement of a tomato plant of the invention with a haploid genetic complement of a second tomato plant, preferably, another, distinct tomato plant. In another aspect, the present invention provides a tomato plant regenerated from a tissue culture that comprises a hybrid genetic complement of this invention.
Any embodiment discussed herein with respect to one aspect of the invention applies to other aspects of the invention as well, unless specifically noted.
The term “about” is used to indicate that a value includes the standard deviation of the mean for the device or method being employed to determine the value. The use of the term “or” in the claims is used to mean “and/or” unless explicitly indicated to refer to alternatives only or the alternatives are mutually exclusive. When used in conjunction with the word “comprising” or other open language in the claims, the words “a” and “an” denote “one or more,” unless specifically noted otherwise. The terms “comprise,” “have” and “include” are open-ended linking verbs. Any forms or tenses of one or more of these verbs, such as “comprises,” “comprising,” “has,” “having,” “includes” and “including,” are also open-ended. For example, any method that “comprises,” “has” or “includes” one or more steps is not limited to possessing only those one or more steps and also covers other unlisted steps. Similarly, any plant that “comprises,” “has” or “includes” one or more traits is not limited to possessing only those one or more traits and covers other unlisted traits.
Other objects, features and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and any specific examples provided, while indicating specific embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.


DETAILED DESCRIPTION OF THE INVENTION
The invention provides methods and compositions relating to plants, seeds and derivatives of tomato hybrid PX 02481245, tomato line PSQ 24-2082 and tomato line PSQ-9Z08005.
Tomato hybrid PX 02481245 is variety for EARLY SEASON THICK segment with an estimated maturity of 118-120 days and capable of EFS (extended field storage). PX 02481245 consistently produces large firm fruit with high viscosity, good color, provides high yield potential with juice bostwick in the 15.5 range and produces thicker viscosity than N6366 and AB2 varieties. AFW is 80 grams and has BRIX of 5.3. This hybrid has good vine structure, vines are uniform and upright, with great bed cover.
PSQ-9Z08005 is jointless, with uniform ripening shoulder, midseason maturity, medium large determinate dark green plant, and medium size square fruit. PSQ-9Z08005 has the ‘alc/nor’ gene so it ripens to apricot/orange color. PSQ-9Z08005 shows resistance to Verticillium (V1), Fusarium (Race 1 & 2) (F1 F2), Nematode, Bacterial Speck Bsk(0).
A. Origin and Breeding History of Tomato Hybrid PX 02481245
The parents of hybrid PX 02481245 are PSQ 24-2082 and PSQ-9Z08005. These parents were created as follows:
Tomato line PSQ 24-2082 was created using a combination of crossing, backcrossing and pedigree selection. All crossing and pedigree selection was done in Seminis greenhouses and field plats in Woodland, Calif. and Meliphilla, Chile.
TMV resistant line 96FB279-5 was used as a source of TMV2a resistance. Two crosses (BC1) to Seminis inbred line 8892/1475 were made, followed by 8 generations of single plant pedigree selection and then bulked in BC1F9. TMV resistance was fixed by PCR Marker in BC1F3. The breeding and selection was carried out as follows:












Year
Generation








Year 1
F1
(96FB279-5 × 8892/1475)


Year 1
BC1
(96FB279-5 × 8892/1475)8892/1475


Year 2
BC1F2


Year 3
BC1F3
Line Fixed for TMV resistance


Year 3
BC1F4


Year 4
BC1F5


Year 4
BC1F6


Year 5
BC1F7


Year 5
BC1F8


Year 6
BC1F9
Line bulked and designated PSQ24-2082 and sent




to Foundation Seed for increase.









Tomato line PSQ-9Z08005 was created using pedigree selection. All pedigree selection was done in field plots in Woodland, Calif. and Meliphilla, Chile.
Accession T7349 was harvested from field in Woodland Calif. and used to create a large segregating population which was planted in Melipilla, Chile. Seven generations of individual plant horticultural pedigree selection followed with the line being bulked in Year 4 for Foundation Seed. The breeding and selection was carried out as follows:












Year
Generation








Year 1
F2
(T7349)


Year 1
F3
(T7349)


Year 2
F4
(T7349)


Year 2
F5
(T7349)


Year 3
F6
(T7349)


Year 3
F7
(T7349)


Year 4
F8
Line (T7349) designated as PSQ-9Z08005 and




given to Foundation Seed for increase.









The parent lines are uniform and stable, as is a hybrid produced therefrom. A small percentage of variants can occur within commercially acceptable limits for almost any characteristic during the course of repeated multiplication. However no variants are expected.
B. Physiological and Morphological Characteristics of Tomato Hybrid PX 02481245, Tomato Line PSQ 24-2082 and Tomato Line PSQ-9Z08005
In accordance with one aspect of the present invention, there is provided a plant having the physiological and morphological characteristics of tomato hybrid PX 02481245 and the parent lines thereof. A description of the physiological and morphological characteristics of such plants is presented in Tables 1-3.







TABLE 1







Physiological and Morphological Characteristics of Hybrid PX 02481245













Comparison Variety:



CHARACTERISTIC
PX 02481245
VF 134














1.
Seedling





Anthocyanin in hypocotyl of
present (Montfavet H 63.4)
present



2-15 cm seedling



Habit of 3-4 week old
normal
normal



seedling


2.
Mature Plant



Height
67.1 cm
68 cm



Growth type
determinate (Campbell
determinate




1327, Prisca)



Only determinate growth
medium (Montfavet H.
medium



type varieties: Plant: number
63.4)



of inflorescences on main



stem (side shoots to be



removed)



Form
normal
lax, open



Size of canopy (compared to
medium
large



others of similar type)



Habit
semi-erect
sprawling


3.
Stem



Anthocyanin coloration of
absent or very weak
absent or very weak



upper third



Branching
intermediate (Westover)
profuse



Branching at cotyledon or
present
present



first leafy node



Number of nodes between
7 to 10
4 to 7



first inflorescence



Number of nodes between
1 to 4
1 to 4



early (1st to 2nd, 2nd to 3rd)



inflorescences



Number of nodes between
1 to 4
1 to 4



later developing



inflorescences



Pubescence on younger stems
sparsely hairy (scattered
moderately hairy




long hairs)


4.
Leaf



Type (mature leaf beneath the
tomato
tomato



3rd inflorescence)



Morphology (mature leaf
Pinnate leaf with medium
Pinnate leaf with small



beneath the 3rd inflorescence)
sized leaflets
sized leaflets



Margins of major leaflets
shallowly toothed or
shallowly toothed or



(mature leaf beneath the 3rd
scalloped
scalloped



inflorescence)



Marginal rolling or wiltiness
slight
absent



(mature leaf beneath the 3rd



inflorescence)



Onset of leaflet rolling
mid season
mid season



(mature leaf beneath the 3rd



inflorescence)



Surface of major leaflets
rugose (bumpy or veiny)
smooth



(mature leaf beneath the 3rd



inflorescence)



Pubescence (mature leaf
normal
hirsute



beneath the 3rd inflorescence)



Attitude (in middle third of
semi-erect (Allround,
horizontal



plant)
Drakar, Vitador)



Length
medium (Lorena)
long



Width
medium
medium



Division of blade
pinnate (Mikado, Pilot, Red
pinnate




Jacket)



Size of leaflets (in middle of
small (Tiny Tim)
large



leaf)



Intensity of green color
dark (Allround, Daniela,
medium




Lorena, Red Robin)



Glossiness (in middle third of
weak (Daniela)
weak



plant)



Blistering (in middle third of
medium (Marmande VR)
none



plant)



Size of blisters (in middle
medium (Marmande VR)
none



third of plant)



Attitude of petiole of leaflet
semi-erect (Blizzard,
semi-drooping



in relation to main axis (in
Marmande VR)



middle third of plant)


5.
Inflorescence



Type (2nd and 3rd truss)
mainly uniparous
mainly multiparous




(Dynamo)



Type (3rd inflorescence)
simple
forked



Average number of flowers
9.6
6.6



in inflorescence (3rd



inflorescence)



Leafy or “running”
absent
occasional



inflorescence (3rd



inflorescence)


6.
Flower



Calyx
normal (lobes awl shaped)
normal



Calyx-lobes
approx. equaling corolla
shorter than corolla



Corolla color
yellow
yellow



Style pubescence
absent or very scarce
sparse




(Campbell 1327)



Anthers
all fused into tube
all fused into tube



Fasciation (1st flower of 2nd
absent (Monalbo,
occasionally present



or 3rd inflorescence)
Moneymaker)



Color
yellow (Marmande VR)
yellow


7.
Fruit



Typical shape in longitudinal
elliptical
circular



section (3rd fruit of 2nd or 3rd



cluster)



Shape of transverse/cross
round
round



section (3rd fruit of 2nd or 3rd



cluster)



Shape of stem end (3rd fruit
indented
flat



of 2nd or 3rd cluster)



Shape of blossom end (3rd
flat to pointed/nippled
flat to pointed/nippled



fruit of 2nd or 3rd cluster)
(Cal J, Early Mech, Peto




Gro)



Size of blossom scar
small (Montfavet H 63.4,
very small




Montfavet H 63.5)



Shape of pistil scar (3rd fruit
DOT
dot



of 2nd or 3rd cluster)



Peduncle: abscission layer
absent (jointless) (Aledo,
present



(3rd fruit of 2nd or 3rd cluster)
Bandera, Count, Lerica)



Ribbing at peduncle end
absent or very weak
absent or very weak




(Calimero, Cerise)



Depression at peduncle end
weak (Futuria, Melody)
absent or very weak



Size of stem/peduncle scar
medium (Montfavet H
small




63.4, Montfavet H 63.5,




Rutgers)



Point of detachment of fruit
at calyx attachment
at pedicel joint



at harvest (3rd fruit of 2nd or



3rd cluster)



Length of mature fruit (stem
58.6 mm
59.35 mm



axis) (3rd fruit of 2nd or 3rd



cluster)



Diameter of fruit at widest
45.7 mm
50.65 mm



point (3rd fruit of 2nd or 3rd



cluster)



Weight of mature fruit (3rd
70 grams
86 grams



fruit of 2nd or 3rd cluster)



Size
medium (Alphamech,
small




Diego)



Ratio length/diameter
medium (Early Mech, Peto
small




Gro)



Core
present
present



Size of core in cross section
medium (Montfavet H
small



(in relation to total diameter)
63.4, Montfavet H 63.5)



Number of locules
3 or 4 (Montfavet H 63.5)
2 or 3



Surface
smooth
smooth



Base color (mature-green
light green (Lanai, VF 145-
apple or medium green



stage)
F5)



Pattern (mature-green stage)
uniform green
green shouldered



Green shoulder (before
absent (Felicia, Rio
absent



maturity)
Grande, Trust)



Intensity of green color of
light (Capello, Duranto,
medium



fruit (before maturity)
Trust)



Color at maturity (full-ripe)
red (Ferline, Daniela,
red




Montfavet H 63.5)



Color of flesh at maturity
red/crimson (Ferline, Saint-
red/crimson



(full-ripe)
Pierre)



Flesh color
with lighter and darker
uniform




areas in walls



Locular gel color of table-
red
red



ripe fruit



Firmness
firm (Femova, Konsul,
medium




Tradior)



Shelf life
long (Daniela)
long



Time of flowering
early (Feria, Primabel)
late



Time of maturity
early (Feria, Rossol)
early



Ripening
uniform
uniform



Ripening
inside out
uniformity



Epidermis color
yellow
yellow



Epidermis
normal
normal



Epidermis texture
average
tough



Thickness of pericarp
medium (Carmello,
medium




Europeel, Floradade, Heinz




1706, Montfavet H 63.5)



Dry matter content (at
high (Aloha, Coudoulet)
medium



maturity)



Sensitivity to silvering
insensitive (Marathon,
insensitive




Sano)


10.
Chemistry and Composition of



Full-ripe Fruits



pH
 4.1
 4.27



Titratable acidity, as % citric
 0.455
 0.3874



Total solids (dry matter,
 7.05
 5.4477



seeds and skins removed)



Soluble solids as °Brix
 6.21
 4.735


11.
Phenology



Seeding to 50% flow (1 open
 55
 71



on 50% of plants)



Seeding to once over harvest
118
118



Fruiting season
very concentrated (UC 82)
medium



Relative maturity in areas
early
early



tested


12.
Adaptation



Culture
field
field



Principle use(s)
whole-pack canning,
whole-pack canning,




concentrated products
concentrated products



Machine harvest
not adapted
not adapted



Regions to which adaptation
Sacramento and Upper San



has been demonstrated
Joaquin Valley of




California





*These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.













TABLE 2







Physiological and Morphological Characteristics of Line PSQ 24-2082










CHARACTERISTIC
PSQ 24-2082













1.
Seedling




Anthocyanin In Hypocotyl Of 2-15 Cm Seedling
Present (Montfavet H 63.4)



Habit Of 3-4 Week Old Seedling
Normal


2.
Mature Plant



Height
58.3 cm



Growth Type
Determinate (Campbell 1327,




Prisca)



Form
Lax, Open



Size Of Canopy (Compared To Others Of
Large



Similar Type)



Habit
Sprawling (Decumbent)


3.
Stem



Branching
Intermediate (Westover)



Branching At Cotyledon Or First Leafy Node
Present



Number Of Nodes Between First Inflorescence
4 To 7



Number Of Nodes Between Early (1st To 2nd, 2nd
1 To 4



To 3rd) Inflorescences



Number Of Nodes Between Later Developing
1 To 4



Inflorescences



Pubescence On Younger Stems
Sparsely Hairy (Scattered Long




Hairs)


4.
Leaf



Type (Mature Leaf Beneath The 3rd
Tomato



Inflorescence)



Margins Of Major Leaflets (Mature Leaf
Shallowly Toothed Or Scalloped



Beneath The 3rd Inflorescence)



Marginal Rolling Or Wiltiness (Mature Leaf
Slight



Beneath The 3rd Inflorescence)



Onset Of Leaflet Rolling (Mature Leaf Beneath
Mid Season



The 3rd Inflorescence)



Surface Of Major Leaflets (Mature Leaf Beneath
Rugose (Bumpy Or Veiny)



The 3rd Inflorescence)



Pubescence (Mature Leaf Beneath The 3rd
Normal



Inflorescence)


5.
Inflorescence



Type (Make Observations On The 3rd
Simple



Inflorescence)



Average Number Of Flowers In Inflorescence
6.2



(Make Observations On The 3rd Inflorescence)



Leafy Or “Running” Inflorescence (Make
Occasional



Observations On The 3rd Inflorescence)


6.
Flower



Calyx
Normal (Lobes Awl Shaped)



Calyx-Lobes
Shorter Than Corolla



Corolla Color
Yellow



Style Pubescence
Absent Or Very Scarce (Campbell




1327)



Anthers
All Fused Into Tube



Fasciation (1st Flower Of 2nd Or 3rd
Absent (Monalbo, Moneymaker)



Inflorescence)


7.
Fruit



Typical Shape In Longitudinal Section (3rd Fruit
Cylindrical



Of 2nd Or 3rd Cluster)



Shape Of Transverse/Cross Section (3rd Fruit
Angular



Of 2nd Or 3rd Cluster)



Shape Of Stem End (3rd Fruit Of 2nd Or 3rd
Flat



Cluster)



Shape Of Blossom End (3rd Fruit Of 2nd Or 3rd
Indented To Flat



Cluster)



Shape Of Pistil Scar (3rd Fruit Of 2nd Or 3rd
Dot



Cluster)



Length Of Mature Fruit (3rd Fruit Of 2nd Or 3rd
63.7 mm



Cluster)



Diameter Of Fruit (3rd Fruit Of 2nd Or 3rd
45.7 mm



Cluster)



Weight Of Mature Fruit (3rd Fruit Of 2nd Or 3rd
76.1 grams



Cluster)



Core
Present



Number Of Locules
2 Or 3 (Alphamech, Futuria)



Surface
Smooth



Base Color (Mature-Green Stage)
Apple Or Medium Green (Heinz




1439 VF)



Pattern (Mature-Green Stage)
Uniform Green



Color At Maturity (Full-Ripe)
Red (Ferline, Daniela, Montfavet H




63.5)



Color Of Flesh At Maturity (Full-Ripe)
Red/Crimson (Ferline, Saint-




Pierre)



Flesh Color
Uniform



Locular Gel Color Of Table-Ripe Fruit
Red



Epidermis Color
Yellow



Epidermis
Easy-Peel



Epidermis Texture
Average



Thickness Of Pericarp
Medium (Carmello, Europeel,




Floradade, Heinz 1706, Montfavet




H 63.5)





*These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.













TABLE 3







Physiological and Morphological Characteristics of Line PSQ-9Z08005













Comparison:



Characteristic
PSQ-9Z08005
PSQ 24-988*HP 988














1.
Seedling





anthocyanin in hypocotyl of 2-15 cm
present
present



seedling
(Montfavet H 63.4)



habit of 3-4 week old seedling
normal
normal


2.
Mature Plant



height
57.2 cm
48.5 cm



growth type
determinate
determinate




(Campbell 1327,




Prisca)



plant: number of inflorescences on
medium (Montfavet
medium



main stem (side shoots to be
H 63.4)



removed)



form
normal
normal



size of canopy (compared to others
medium
small



of similar type)



habit
semi-erect
semi-erect


3.
Stem



anthocyanin coloration of upper third
absent or very weak
absent or very weak



branching
intermediate
intermediate




(Westover)



branching at cotyledon or first leafy
absent
present



node



number of nodes between first
7 to 10
4 to 7



inflorescence



number of nodes between early (1st
1 to 4
1 to 4



to 2nd, 2nd to 3rd) inflorescences



number of nodes between later
1 to 4
1 to 4



developing inflorescences



pubescence on younger stems
sparsely hairy
moderately hairy




(scattered long hairs)


4.
Leaf



type (mature leaf beneath the 3rd
tomato
tomato



inflorescence)



margins of major leaflets (mature
shallowly toothed or
shallowly toothed or



leaf beneath the 3rd inflorescence)
scalloped
scalloped



marginal rolling or wiltiness (mature
moderate
moderate



leaf beneath the 3rd inflorescence)



onset of leaflet rolling
mid season
mid season



surface of major leaflets (mature leaf
rugose (bumpy or
rugose



beneath the 3rd inflorescence)
veiny)



pubescence (mature leaf beneath the
normal
normal



3rd inflorescence)



attitude (in middle third of plant)
semi-erect (Allround,
semi-erect




Drakar, Vitador)



length
medium (Lorena)
short



width
medium
narrow



division of blade
pinnate (Mikado,
pinnate




Pilot, Red Jacket)



size of leaflets (in middle of leaf)
small (Tiny Tim)
small



intensity of green color
dark (Allround,
dark




Daniela, Lorena, Red




Robin)



glossiness (as for 6)
weak (Daniela)
medium



blistering (as for 6)
medium
strong




(Marmande VR)



size of blisters (as for 6)
medium
medium




(Marmande VR)



attitude of petiole of leaflet in
semi-erect (Blizzard,
horizontal



relation to main axis (in middle of
Marmande VR)



leaf)


5.
Inflorescence



inflorescence type (2nd and 3rd truss)
mainly uniparous
mainly uniparous




(Dynamo)



type (3rd inflorescence)
simple
simple



average number of flowers in
7.2
5.7



inflorescence (3rd inflorescence)



leafy or “running” inflorescence (3rd
absent
absent



inflorescence)


6.
Flower



calyx
normal (lobes awl
normal




shaped)



calyx-lobes
shorter than corolla
shorter than corolla



corolla color
yellow
yellow



style pubescence
absent or very scarce
absent or very scarce




(Campbell 1327)



anthers
all fused into tube
all fused into tube



fasciation (1st flower of 2nd or 3rd
absent (Monalbo,
absent



inflorescence)
Moneymaker)



color
yellow
yellow




(Marmande VR)


7.
Fruit



typical shape in longitudinal section
elliptical
elliptical



(3rd fruit of 2nd or 3rd cluster)



shape of transverse/cross section
round
round



(3rd fruit of 2nd or 3rd cluster)



shape of stem end (3rd fruit of 2nd or
flat
flat



3rd cluster)



shape of blossom end (3rd fruit of 2nd
flat to pointed/
flat to pointed/



or 3rd cluster)
nippled (Cal J, Early
nippled




Mech, Peto Gro)



size of blossom scar
medium (Alphamech,
small




Apla, Carmello,




Floradade)



shape of pistil scar (3rd fruit of 2nd or
stellate
dot



3rd cluster)



peduncle: abscission layer (3rd fruit
absent (jointless)
absent



of 2nd or 3rd cluster)
(Aledo, Bandera,




Count, Lerica)



ribbing at peduncle end
medium
weak




(Montfavet H 63.4,




Montfavet H 63.5)



depression at peduncle end
absent or very weak
absent or very weak




(Europeel, Heinz




1706, Rossol, Sweet




Baby)



size of stem/peduncle scar
medium
small




(Montfavet H 63.4,




Montfavet H 63.5,




Rutgers)



point of detachment of fruit at
at calyx attachment
at calyx attachment



harvest (3rd fruit of 2nd or 3rd cluster)



length of mature fruit (stem axis; 3rd
64.7 mm
52.3 mm



fruit of 2nd or 3rd cluster)



diameter of fruit at widest point (3rd
48.3 mm
41.1 mm



fruit of 2nd or 3rd cluster)



weight of mature fruit (3rd fruit of 2nd
84.6 grams
49.2 grams



or 3rd cluster)



size
medium (Alphamech,
small




Diego)



ratio length/diameter
medium (Early Mech,
medium




Peto Gro)



core
present
present



size of core in cross section (in
medium
medium



relation to total diameter)
(Montfavet H 63.4,




Montfavet H 63.5)



number of locules
3 or 4
2 or 3




(Montfavet H 63.5)



surface
smooth
smooth



base color (mature-green stage)
light green (Lanai,
apple or medium green




VF 145-F5)



pattern (mature-green stage)
uniform green
uniform green



green shoulder (before maturity)
absent (Felicia, Rio
absent




Grande, Trust)



color at maturity (full-ripe)
red (Ferline, Daniela,
red




Montfavet H 63.5)



color of flesh at maturity (full-ripe)
red/crimson
red crimson




(Ferline, Saint-Pierre)



flesh color
uniform
uniform



locular gel color of table-ripe fruit
red
red



firmness
firm (Fernova,
firm




Konsul, Tradiro)



shelf life
medium (Durinta)
medium



time of flowering
medium
medium




(Montfavet H 63.5,




Prisca)



time of maturity
late (Manific, Saint-
medium




Pierre)



ripening (blossom-to-stem axis)
uniform
blossom to stem end



ripening (peripheral to central radial
uniformity
uniformity



axis)



epidermis color
yellow
yellow



epidermis
normal
normal



epidermis texture
tender
tender



thickness of pericarp
thick (Cal J, Daniela,
thick




Ferline, Peto Gro,




Rio Grande)



dry matter content (at maturity)
medium
medium



sensitivity to silvering
insensitive
insensitive




(Marathon, Sano)


8.
Chemistry and Composition of Full-Ripe



Fruits



pH
4.15
4.24



titratable acidity, as % citric
0.517
0.45



total solids (dry matter, seeds and
8.2
6.5



skin removed)



soluble Solids as °Brix
7.5
5.8


9.
Phenology



seeding to 50% flow (1 open on
55 days
55 days



50% of plants)



seeding to once over harvest (if
153 days
132 days



applicable)



fruiting season
medium (Westover)
medium



relative maturity in areas tested
medium
medium


10.
Adaptation



culture
field
field





*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 Tomato Plants
One aspect of the current invention concerns methods for producing seed of tomato hybrid PX 02481245 involving crossing tomato lines PSQ 24-2082 and PSQ-9Z08005. Alternatively, in other embodiments of the invention, hybrid PX 02481245, line PSQ 24-2082, or line PSQ-9Z08005 may be crossed with itself or with any second plant. Such methods can be used for propagation of hybrid PX 02481245 and/or the tomato lines PSQ 24-2082 and PSQ-9Z08005, or can be used to produce plants that are derived from hybrid PX 02481245 and/or the tomato lines PSQ 24-2082 and PSQ-9Z08005. Plants derived from hybrid PX 02481245 and/or the tomato lines PSQ 24-2082 and PSQ-9Z08005 may be used, in certain embodiments, for the development of new tomato varieties.
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 PX 02481245 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.
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.
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.
The plants 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 PX 02481245 and/or tomato lines PSQ 24-2082 and PSQ-9Z08005 for the purpose of developing novel tomato 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 fruit yield, disease tolerance or resistance, and adaptability for soil and climate conditions. Consumer-driven traits, such as a fruit shape, color, texture, and taste are other examples of traits that may be incorporated into new lines of tomato plants developed by this invention.
D. Performance Characteristics
As described above, hybrid PX 02481245 exhibits desirable traits, as conferred by tomato lines PSQ 24-2082 and PSQ-9Z08005. The performance characteristics of hybrid PX 02481245 and tomato lines PSQ 24-2082 and PSQ-9Z08005 were the subject of an objective analysis of the performance traits relative to other varieties. The results of the analysis are presented below.







TABLE 4







Performance Characteristics for Hybrid PX 02481245 and Comparative varieties





















Disease

Fruit
Wall
Brix
Jbost

Ostwald







Variety
Resistance
Days
Wt (g)
(mm)
(%)
(cm)
PPB
(s)
a/b
pH
MA
SAR
Lycopene





S 6366
VF2NPst(0)
118
84.6
7.5
5.4
20.1
7.5
197
2.37
4.51
16.0
7.5
119


AB 0311
VF2NPst(0)
118
89.5
7.7
5.6
17.4
6.5
216
2.40
4.38
19.9
5.9
122



TSWV


AB 2
VF2Pst(0)
122
95.6
8.1
5.3
19.3
7.0
209
2.37
4.38
19.6
6.0
111


AB 3
VF2NPst(0)
122
99.3
8.1
5.4
19.1
7.0
185
2.35
4.41
18.1
6.5
113


PS 024 7
VF2N TSWV
125
93.7
8.4
5.0
15.8
4.7
280
2.39
4.44
18.4
6.4
123


0002


PX 024 8
VF2NPst(0)
125
87.6
6.9
5.2
16.5
5.3
293
2.27
4.35
20.7
5.7
100


1245
EFS


CXD255
VF2NPst(0)
125
92.9
8.0
5.2
14.7
4.3
348
2.35
4.40
17.9
6.5
119



EFS


Hypeel 849
VF2NPst(0)
128
87.3
8.1
4.9
15.4
4.4
393
2.35
4.36
20.4
5.8
110



EFS


PS 024 5
VF2NPst(0)
130
94.6
8.3
5.3
15.2
4.8
263
2.33
4.46
17.4
6.8
101


0650
EFS


H5608
VF2NPst(0)
128
76.4
7.3
5.0
12.5
3.0
489
2.51
4.39
20.4
5.8
131



TSWV





*** Key for Table 4:


% Brix Raw puree solids based on refractive index


Juice Bostwick (Jbost) Hot Break puree flow in centimeters/30 seconds − average of 2 samples (lower value indicates higher viscosity)


Predicted Paste Bostwick (PPB) Estimated Bostwick of 12% Brix paste, calculated using Brix and Juice Bostwick values (lower value indicates higher viscosity)


Ostwald Serum viscosity of hot break puree serum measured in seconds of flow at 30 C., relative to water (higher number indicates higher viscosity)


pH Standard pH values of raw puree


Sugar to Acid ratio (SAR) Ratio based on total solids and titratable acidity.


Master Acid (MA) Acidity calculation based on Titratable Acidity and Total Solids


a/b Color (a/b) Represents the hue or color of the sample using a Minolta CM-508d (higher number is “redder” color)


Lycopene Lycopene (ppm) in raw puree


Fruit Weight Average weight of 10 fruit, measured in gram


Fruit Wall (Wall) Average width of pericarp wall of 10 fruit, measured in millimeters


Disese Resistance V—Verticillium; F1—Fusarium (Race 1); F2—Fusarium (Race 2); N—Nematode; BSK—Bacterial Speck













TABLE 5







Further Performance Characteristics For Hybrid PX 02481245










Variety
Tons/Acre







PX 02481245 (A)
60.2



FV (A)
62.0



S 6366 (A, B)
59.2



AB2 (B, C)
55.6



H 8504 (C)
54.4







** The results shown in Table 5 are taken from three trial locations in the same growing season. “A” and “B” indicate varieties whose production is not statistically different from each other. Varieties not connected by the same letter are significantly different.






E. Further Embodiments of the Invention
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 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 tomato plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the 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 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.
Backcrossing methods can be used with the present invention to improve or introduce a characteristic into the present variety. The parental tomato 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 tomato 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.
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 tomato plant is obtained wherein essentially all of the 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.
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.
In one embodiment, progeny tomato plants of a backcross in which a plant described herein is the recurrent parent comprise (i) the desired trait from the non-recurrent parent and (ii) all of the physiological and morphological characteristics of tomato the recurrent parent as determined at the 5% significance level when grown in the same environmental conditions.
New 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.
With the development of molecular markers associated with particular traits, it is possible to add additional traits into an established germ line, such as represented here, with the end result being substantially the same base germplasm with the addition of a new trait or traits. Molecular breeding, as described in Moose and Mumm, 2008 (Plant Physiology, 147: 969-977), for example, and elsewhere, provides a mechanism for integrating single or multiple traits or QTL into an elite line. This molecular breeding-facilitated movement of a trait or traits into an elite line may encompass incorporation of a particular genomic fragment associated with a particular trait of interest into the elite line by the mechanism of identification of the integrated genomic fragment with the use of flanking or associated marker assays. In the embodiment represented here, one, two, three or four genomic loci, for example, may be integrated into an elite line via this methodology. When this elite line containing the additional loci is further crossed with another parental elite line to produce hybrid offspring, it is possible to then incorporate at least eight separate additional loci into the hybrid. These additional loci may confer, for example, such traits as a disease resistance or a fruit quality trait. In one embodiment, each locus may confer a separate trait. In another embodiment, loci may need to be homozygous and exist in each parent line to confer a trait in the hybrid. In yet another embodiment, multiple loci may be combined to confer a single robust phenotype of a desired trait.
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.
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.
Selection of tomato 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., Nucleic Acids Res., 1 8:6531-6535, 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., Science, 280:1077-1082, 1998).
F. Plants Derived by Genetic Engineering
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.
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.
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.
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. Microprojectile bombardment techniques are widely applicable, and may be used to transform virtually any plant species.
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., Bio-Technology, 3(7):637-642, 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.
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., Bio/Technology, 3:629-635, 1985; U.S. Pat. No. 5,563,055).
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., Mol. Gen. Genet., 199:183-188, 1985; Omirulleh et al., Plant Mol. Biol., 21(3):415-428, 1993; Fromm et al., Nature, 312:791-793, 1986; Uchimiya et al., Mol. Gen. Genet., 204:204, 1986; Marcotte et al., Nature, 335:454, 1988). Transformation of plants and expression of foreign genetic elements is exemplified in Choi et al. (Plant Cell Rep., 13: 344-348, 1994), and Ellul et al. (Theor. Appl. Genet., 107:462-469, 2003).
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 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., Nature, 313:810, 1985), including in monocots (see, e.g., Dekeyser et al., Plant Cell, 2:591, 1990; Terada and Shimamoto, Mol. Gen. Genet., 220:389, 1990); a tandemly duplicated version of the CaMV 35S promoter, the enhanced 35S promoter (P-e35S); 1 the nopaline synthase promoter (An et al., Plant Physiol., 88:547, 1988); the octopine synthase promoter (Fromm et al., Plant Cell, 1:977, 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.
A variety of plant gene promoters that are regulated in response to environmental, hormonal, chemical, and/or developmental signals can also be used for expression of an operably linked gene in plant cells, including promoters regulated by (1) heat (Callis et al., Plant Physiol., 88:965, 1988), (2) light (e.g., pea rbcS-3A promoter, Kuhlemeier et al., Plant Cell, 1:471, 1989; maize rbcS promoter, Schaffner and Sheen, Plant Cell, 3:997, 1991; or chlorophyll a/b-binding protein promoter, Simpson et al., EMBO J., 4:2723, 1985), (3) hormones, such as abscisic acid (Marcotte et al., Plant Cell, 1:969, 1989), (4) wounding (e.g., wunl, Siebertz et al., Plant Cell, 1:961, 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., EMBO J., 6:1155, 1987; Schernthaner et al., EMBO J., 7:1249, 1988; Bustos et al., Plant Cell, 1:839, 1989).
Exemplary nucleic acids which may be introduced to 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.
Many hundreds if not thousands of different genes are known and could potentially be introduced into a tomato plant according to the invention. Non-limiting examples of particular genes and corresponding phenotypes one may choose to introduce into a tomato 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 in 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.
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., Biotech. Gen. Engin. Rev., 9:207, 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, Mol. Biotech., 7:125, 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
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:
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.
Backcrossing: A process in which a breeder repeatedly crosses hybrid progeny, for example a first generation hybrid (F1), 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.
Crossing: The mating of two parent plants.
Cross-pollination: Fertilization by the union of two gametes from different plants.
Diploid: A cell or organism having two sets of chromosomes.
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.
Enzymes: Molecules which can act as catalysts in biological reactions.
F1 Hybrid: The first generation progeny of the cross of two nonisogenic plants.
Genotype: The genetic constitution of a cell or organism.
Haploid: A cell or organism having one set of the two sets of chromosomes in a diploid.
Linkage: A phenomenon wherein alleles on the same chromosome tend to segregate together more often than expected by chance if their transmission was independent.
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.
Phenotype: The detectable characteristics of a cell or organism, which characteristics are the manifestation of gene expression.
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.
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.
Regeneration: The development of a plant from tissue culture.
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.
Self-pollination: The transfer of pollen from the anther to the stigma of the same plant.
Single Locus Converted (Conversion) Plant: Plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a tomato 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.
Substantially Equivalent: A characteristic that, when compared, does not show a statistically significant difference (e.g., p=0.05) from the mean.
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.
Transgene: A genetic locus comprising a sequence which has been introduced into the genome of a tomato plant by transformation.
H. Deposit Information
A deposit of tomato hybrid PX 02481245 and inbred parent lines PSQ 24-2082 and PSQ-9Z08005, 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 deposits were Dec. 14, 2011, Feb. 5, 2009, and Dec. 14, 2011, respectively. The accession numbers for those deposited seeds of tomato hybrid PX 02481245 and inbred parent lines PSQ 24-2082 and PSQ-9Z08005 are ATCC Accession No. PTA-12335, ATCC Accession No. PTA-9756, and ATCC Accession No. PTA-12336, 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.
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.
All references cited herein are hereby expressly incorporated herein by reference.

## Claims

1. A tomato plant of hybrid PX 02481245, a sample of seed of said hybrid having been deposited under ATCC Accession Number PTA-12335.

2. A seed of hybrid PX 02481245, a sample of seed of said hybrid having been deposited under ATCC Accession Number PTA-12335.

3. A seed that produces the plant of claim 1.

4. A plant part of the plant of claim 1.

5. The plant part of claim 4, further defined as a leaf, an ovule, pollen, a fruit, or a cell.

6. A tissue culture of regenerable cells of the plant of claim 1.

7. The tissue culture according to claim 6, comprising cells or protoplasts from a plant part selected from the group consisting of embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistil, flower, seed and stalks, wherein the plant part comprises a cell of tomato hybrid PX 02481245.

8. A tomato plant regenerated from the tissue culture of claim 6, wherein said plant has all of the physiological and morphological characteristics of tomato hybrid PX 02481245, a sample of seed of said hybrid having been deposited under ATCC Accession Number PTA-12335.

9. A method of vegetatively propagating the plant of claim 1 comprising the steps of:
(a) collecting tissue capable of being propagated from the plant according to claim 1;
(b) cultivating said tissue to obtain proliferated shoots; and
(c) rooting said proliferated shoots to obtain rooted plantlets.

10. The method of claim 9, further comprising growing at least a first plant from said rooted plantlets.

11. A method of introducing a desired trait into tomato hybrid PX 02481245 comprising:
(a) introducing at least a first heritable trait into a tomato line selected from the group consisting of line PSQ 24-2082 and line PSQ-9Z08005 to produce a plant of the first inbred tomato line that heritably carries the trait, wherein the heritable trait is introduced into said first tomato line by backcrossing and wherein representative samples of seed of tomato line PSQ 24-2082 and line PSQ-9Z08005 have been deposited under ATCC Accession Number PTA-9756, and ATCC Accession Number PTA-12336, respectively; and
(b) crossing a plant of the first inbred tomato line that heritably carries the trait with a plant of a different line selected from said group consisting of PSQ 24-2082 and line PSQ-9Z08005 to produce a plant of hybrid PX 02481245 comprising the heritable trait.

12. A tomato plant produced by the method of claim 11, wherein the plant comprises the desired trait and otherwise comprises essentially all of the morphological and phenotypic traits of hybrid PX 02481245, a sample of seed of said hybrid having been deposited under ATCC Accession Number PTA-12335.

13. A method of producing a plant comprising an added trait, the method comprising introducing a transgene conferring the trait into a plant of hybrid PX 02481245, a sample of seed of said hybrid having been deposited under ATCC Accession Number PTA-12335.

14. A plant produced by the method of claim 13.

15. The plant of claim 1, further comprising a transgene.

16. The plant of claim 15, wherein the transgene confers a trait selected from the group consisting of male sterility, herbicide tolerance, insect resistance, pest resistance, disease resistance, modified fatty acid metabolism, environmental stress tolerance, modified carbohydrate metabolism and modified protein metabolism.

17. The plant of claim 1, further comprising a single locus conversion, a sample of seed of said hybrid having been deposited under ATCC Accession Number PTA-12335, wherein said plant otherwise comprises essentially all of the morphological and physiological characteristics of tomato hybrid PX 02481245.

18. The plant of claim 17, wherein the single locus conversion confers a trait selected from the group consisting of male sterility, herbicide tolerance, insect resistance, pest resistance, disease resistance, modified fatty acid metabolism, environmental stress tolerance, modified carbohydrate metabolism and modified protein metabolism.

19. A method for producing a seed of a plant derived from hybrid PX 02481245 comprising the steps of:
(a) crossing a tomato plant of hybrid PX 02481245 with itself or a second tomato plant; a sample of seed of said hybrid having been deposited under ATCC Accession Number PTA-12335; and
(b) allowing seed of a hybrid PX 02481245-derived tomato plant to form.

20. The method of claim 19, further comprising the steps of:
(c) selfing a plant grown from said hybrid PX 02481245-derived tomato seed to yield additional hybrid PX 02481245-derived tomato seed;
(d) growing said additional hybrid PX 02481245-derived tomato seed of step (c) to yield additional hybrid PX 02481245-derived tomato plants; and
(e) repeating the selfing and growing steps of (c) and (d) to generate at least a first further hybrid PX 02481245-derived tomato plant.

21. The method of claim 19, wherein the second tomato plant is of an inbred tomato line.

22. The method of claim 20, further comprising:
(f) crossing the further hybrid PX 02481245-derived tomato plant with a different tomato plant to produce seed of a hybrid progeny plant.

23. A method of producing a tomato seed comprising crossing the plant of claim 1 with itself or a second tomato plant and allowing seed to form.

24. A method of producing a tomato fruit comprising:
(a) obtaining the plant according to claim 1, wherein the plant has been cultivated to maturity; and
(b) collecting a tomato from the plant.