Hybrid mushroom strain J11500 and descendants thereof

Abstract

A hybrid mushroom culture of Agaricus bisporus, designated as strain J11500, includes a representative culture of the strain, which has been deposited under NRRL Accession No. 50895. A method of producing a hybrid mushroom culture of Agaricus bisporus comprising: mating a homokaryotic line J10102-s69 with a homokaryotic line OWNC. Additionally, mushrooms, parts of the culture and products incorporating the culture are provided.

Claims

1. A hybrid mushroom culture of Agaricus bisporus designated as strain J11500, a representative culture of the strain having been deposited under NRRL Accession No. 50895.

2. A part of the hybrid mushroom culture of claim 1 selected from the group consisting of hyphae, spores, cells, nuclei, and protoplasts.

3. The part of the hybrid mushroom culture of claim 2, wherein the spores are selected from dormant and germinated spores, and wherein the dormant and germinated spores include heterokaryons and homokaryons incorporated therein.

4. A product comprising the hybrid mushroom culture of Agaricus bisporus designated as strain J11500 of claim 1, the product selected from the group consisting of mycelium, spawn, inoculum, casing inoculum, fresh mushrooms, processed mushrooms, mushroom pieces, and colonized substrates including grain, compost, and friable particulate matter.

5. An Essentially Derived Variety of the hybrid mushroom culture of claim 1, wherein said Essentially Derived Variety is a culture of a strain derived from a single initial culture of strain J11500, wherein a culture of the strain has been deposited under NRRL Accession No. 50895, such that at least 75% of its genome or genotype is present in the genome or genotype of the initial culture of strain J11500.

6. A hybrid mushroom culture of Agaricus bisporus having a genotypic fingerprint which has alleles at marker loci ITS, p1n150-G3-2, MFPC-1-ELF, AN, AS, and FF, wherein all of the alleles at marker loci ITS, p1n150-G3-2, MFPC-1-ELF, AN, AS, and FF of said fingerprint are present in the genotypic fingerprint of strain J11500, wherein a culture of strain J11500 has been deposited under NRRL Accession No. 50895.

7. The hybrid mushroom culture of claim 6, wherein said culture has a genotypic fingerprint which has characters at marker loci described in Table VII, wherein all of the characters of said fingerprint are present in the genotypic fingerprint of strain J11500, wherein a culture of strain J11500 has been deposited under NRRL Accession No. 50895.

8. An Agaricus bisporus culture having all of the physiological and morphological characteristics of strain J11500, wherein a culture of strain J11500 has been deposited under the NRRL Accession Number 50895.

9. A method of producing a hybrid mushroom culture of Agaricus bisporus comprising: mating a homokaryotic line J10102-s69, a culture of which has been deposited under NRRL Accession No. 50893, with a homokaryotic line OWNC, a culture of which has deposited under NRRL Accession No. 50894.

10. The method of claim 9, wherein said hybrid mushroom culture exhibits antagonism toward heterokaryon strains in the U1 derived lineage group.

11. The method according to claim 9, further comprising: providing the mushroom culture in mushroom products selected from the group consisting of mycelium, spawn, inoculum, casing inoculum, fresh mushrooms, processed mushrooms, parts of mushrooms, mushroom pieces, and colonized substrates including grain, compost, and friable particulate matter.

12. The method according to claim 9, further comprising: providing the mushroom culture in derived cultures selected from the group consisting of homokaryons, heterokaryons, aneuploids, somatic subcultures, tissue explants cultures, protoplasts, dormant spores, germinating spores, inbred descendents and outbred descendents, transgenic cultures, and cultures having a genome incorporating a single locus conversion.

13. A culture produced by the method of claim 9.

14. The method of claim 10, further comprising growing the hybrid mushroom culture to produce hybrid mushrooms and parts of mushrooms.

15. The method of claim 9, wherein the hybrid mushroom culture produced comprises a marker profile having alleles at marker loci ITS, p1n150-G3-2, MFPC-1-ELF, AN, AS, and FF, wherein all of the alleles at marker loci ITS, p1n150-G3-2 MFPC-1-ELF, AN, AS, and FF of said marker profile are also present in the marker profile of J11500.

16. A cell of the hybrid culture produced by the method of claim 9.

17. The cell of claim 16, further comprising a marker profile having alleles at marker loci ITS, p1n150-G3-2, MFPC-1-ELF, AN, AS, and FF, wherein all of the alleles at marker loci ITS, p1n150-G3-2 MFPC-1-ELF, AN, AS, and FF of said marker profile are also present in the marker profile of J11500.

18. The cell of claim 17, further comprising a marker profile having characters at marker loci described in Table VII, wherein all of the characters of said marker profile are also present in the marker profile of J11500.

19. A culture comprising the cell of claim 16.

20. A method comprising using a hybrid mushroom culture selected from strain J11500 or Essentially Derived Varieties of strain J11500, wherein said Essentially Derived Varieties are cultures of a strain derived from a single initial culture of strain J11500, such that at least 75% of its genome or genotype is present in the genome or genotype of the initial culture of strain J11500, a representative culture of the strain J11500 having been deposited under NRRL Accession No. 50895.

21. The method of claim 20, further comprising: growing a crop of edible mushrooms.

22. The method of claim 20, further comprising: using strain J11500 or Essentially Derived Varieties of strain J11500 in crop rotation to reduce pathogen pressure and pathogen reservoirs in mushroom growing facilities.

23. The method of claim 20, further comprising using strain J11500 and Essentially Derived Varieties of strain J11500 to produce offspring.

Description

DETAILED DESCRIPTION OF THE INVENTION

(1) Initially, in order to provide clear and consistent understanding of the specification and claims, including the scope to be given such terms, the following definitions are provided.

(2) Allele: A heritable unit of the genome at a defined locus, ultimately identified by its DNA sequence (or by other means).

(3) Amphithallism: A reproductive syndrome in which heteromixis and intramixis are both active.

(4) Anastomosis: Fusion of two or more hyphae that achieves cytoplasmic continuity.

(5) Basidiomycete: A monophyletic group of fungi producing meiospores on basidia; a member of a corresponding subdivision of Fungi such as the Basidiomycetales or Basidiomycotina.

(6) Basidium: The meiosporangial cell, in which karyogamy and meiosis occur, and upon which the basidiospores are formed.

(7) Bioefficiency: For mushroom crops, the net fresh weight of the harvested crop divided by the dry weight of the compost substrate at the time of spawning, for any given sampled crop area or compost weight.

(8) Breeding: Development of strains, lines or varieties using methods that emphasize sexual mating.

(9) Cap: Pileus; part of the mushroom, the gill-bearing structure.

(10) Cap Roundness: Strictly, a ratio of the maximum distance between the uppermost and lowermost parts of the cap, divided by the maximum distance across the cap, measured on a longitudinally bisected mushroom; typically averaged over many specimens; subjectively, a rounded property of the shape of the cap.

(11) Carrier substrate: A medium having both nutritional and physical properties suitable for achieving both growth and dispersal of a culture.

(12) Casing layer, casing: A layer of non-nutritive material such as peat or soil that is applied to the upper surface of a mass of colonized compost in order to permit development of the mushroom crop.

(13) Casing inoculum (CI): A formulation of inoculum material incorporating a mushroom culture, typically of a defined heterokaryotic strain, suitable for mixing into the casing layer.

(14) Cloning: Somatic propagation without selection.

(15) Combining ability: The capacity of an individual to transmit traits or superior performance to its offspring (known and available methods of assessment vary by trait).

(16) Compatibility: See heterokaryon compatibility.

(17) Culture: The tangible living organism; the organism propagated on various growth media and substrates; one instance of one physical strain, line, homokaryon or heterokaryon; the sum of all of the parts of the culture, including hyphae, mushrooms, spores, cells, nuclei, mitochondria, cytoplasm, protoplasts, DNA, RNA, proteins, cell membranes and cell walls.

(18) Derivation: Development from a strain; see Essentially Derived Variety (EDV).

(19) Derived lineage group: The set of EDVs derived from a single initial strain or variety.

(20) Descent: Genealogical descent over a limited number (e.g., 10 or fewer) of generations.

(21) Diploid: Having two haploid chromosomal complements within a single nuclear envelope.

(22) Essential derivation: A process by which an Essentially Derived Variety is obtained from an initial variety or strain or from an EDV of an initial variety or strain; modification of an initial culture using methods including somatic selection, tissue culture selection, selfing including intramictic reproduction via single spores and multiple spores and mating of sibling offspring lines, back-mating to the initial variety, or mutagenesis and/or genetic transformation of the initial variety to produce a distinct culture in which the genotype of the resulting culture is predominantly that of the initial culture.

(23) Essentially Derived Variety (EDV): (Note: EDV definitions incorporate elements of (1) relatedness, (2) methods of derivation, (3) and empirical tests.) In general, a variety that is predominantly derived from an initial variety or from an EDV of an initial variety, and which conforms to essential characteristics of the initial variety except for distinguishing differences resulting from the act of derivation, is an EDV of the initial variety. In the art of mushroom strain development, a strain or culture predominantly or entirely derived from a single initial strain or culture, thus having most or all, but at least 75%, of its genome or genotype present in the genome or genotype of the initial strain or culture; a strain or culture obtained from an initial strain or culture by somatic selection, tissue culture selection, selfing including mating of sibling offspring lines and intramictic reproduction via single or multiple spores, back-mating to the initial strain or culture, or mutagenesis and/or genetic transformation of the initial strain or culture; a strain or culture reconstituted from neohaplonts derived from an initial strain or culture, whether or not the haploid lines have been passed into or out of other heterokaryons; a strain or culture with the same essential phenotype as that of an initial strain or culture.

(24) Flesh Thickness: A ratio of the maximum distance between the top of the stem and the uppermost part of the cap, divided by the maximum distance across the cap, measured on a longitudinally bisected mushroom; typically averaged over many specimens; subjectively called meatiness.

(25) Flush: A period of mushroom production within a cropping cycle, separated by intervals of non-production; the term flush encompasses the terms break and wave and can be read as either of those terms.

(26) Fungus: An organism classified as a member of the Kingdom Fungi.

(27) Genotypic fingerprint: A description of the genotype at a defined set of marker loci; the known genotype.

(28) Gill: Lamella; part of the mushroom, the hymenophore- and basidium-bearing structure.

(29) Haploid: Having only a single complement of nuclear chromosomes; see homokaryon.

(30) Heteroallelic: Having two different alleles at a locus; analogous to heterozygous.

(31) Heteroallelism: Differences between homologous chromosomes in a heterokaryotic genotype; analogous to heterozygosity.

(32) Heterokaryon: As a term of art this refers to a sexual heterokaryon: a culture which has two complementary (i.e., necessarily heteroallelic at the Mat locus) types of haploid nuclei in a common cytoplasm, and is thus functionally and physiologically analogous to a diploid individual (but cytogenetically represented as N+N rather than 2N), and which is potentially reproductively competent, and which exhibits self/non-self incompatibility reactions with other heterokaryons; also called a strain or stock in the breeding context.

(33) Heterokaryon compatibility: The absence of antagonism observed during physical proximity or contact between two heterokaryons that are not genetically identical; see Heterokaryon Incompatibility.

(34) Heterokaryon incompatibility: The phenomenon of antagonism observed during physical proximity or contact between two heterokaryons that are not genetically identical; a multilocus self/non-self recognition system that operates in basidiomycete heterokaryons.

(35) Heterokaryotic: Having the character of a heterokaryon.

(36) Heteromixis: Life cycle involving mating between two different non-sibling haploid individuals or gametes; outbreeding.

(37) Homoallelic: Having not more than one allele at a locus. The equivalent term in a diploid organism is homozygous. Haploid lines are by definition entirely homoallelic at all non-duplicated loci.

(38) Homokaryon: A haploid culture with a single type (or somatic lineage) of haploid nucleus (cytogenetically represented as N), and which is ordinarily reproductively incompetent, and which does not exhibit typical self/non-self incompatibility reactions with heterokaryons, and which may function as a gamete in sexually complementary anastomoses; a line which, as with an inbred plant line, transmits a uniform genotype to offspring; a predominantly homoallelic line that mates well and fruits poorly is a putative homokaryon for strain development purposes; see discussion below.

(39) Homokaryotic: Having the character of a homokaryon; haploid.

(40) Hybrid: Of biparental origin, usually applied to heterokaryotic strains and cultures produced in controlled matings.

(41) Hybridizing: Physical association, for example on a petri dish containing a sterile agar-based nutrient medium, of two cultures, usually homokaryons, in an attempt to achieve anastomosis, plasmogamy, and formation of a sexual heterokaryon (=mating); succeeding in the foregoing.

(42) Hyphae: Threadlike elements of mycelium, composed of cell-like compartments.

(43) Inbreeding: Matings that include sibling-line matings, back-matings to parent lines or strains, and intramixis; reproduction involving parents that are genetically related.

(44) Incompatibility: See heterokaryon incompatibility.

(45) Inoculum: A culture in a form that permits transmission and propagation of the culture, for example onto new media; specialized commercial types of inoculum include spawn and CI.

(46) Intramixis: A uniparental sexual life cycle involving formation of a complementary mated pair of postmeiotic nuclei within the basidium or individual spore.

(47) Introgressive trait conversion: mating offspring of a hybrid to a parent line or strain such that a desired trait from one strain is introduced into a predominating genetic background of the other parent line or strain.

(48) Lamella: see gill.

(49) Line: A culture used in matings to produce a hybrid strain; ordinarily a homokaryon which is thus homoallelic, otherwise a non-heterokaryotic (non-NSNPP) culture which is highly homoallelic; practically, a functionally homokaryotic and entirely or predominantly homoallelic culture; analogous in plant breeding to an inbred line which is predominantly or entirely homozygous.

(50) Lineage group: see derived lineage group. The set of EDVs derived from a single initial strain or variety.

(51) Locus: A defined contiguous part of the genome, homologous although often varying among different genotypes; plural: loci.

(52) Marker assisted selection: Using linked genetic markers including molecular markers to track trait-determining loci of interest among offspring and through pedigrees.

(53) MAT: The mating-type locus, which determines sexual compatibility and the heterokaryotic state.

(54) Mating: The sexual union of two cultures via anastomosis and plasmogamy; methods of obtaining matings between mushroom cultures are well known in the art.

(55) Mycelium: The vegetative body or thallus of the mushroom organism, comprised of threadlike hyphae.

(56) Mushroom: The reproductive structure of an agaric fungus; an agaric; a cultivated food product of the same name.

(57) Neohaplont: A haploid culture or line obtained by physically deheterokaryotizing (reducing to haploid components) a heterokaryon; a somatically obtained homokaryon.

(58) Offspring: Descendents, for example of a parent heterokaryon, within a single generation; most often used to describe cultures obtained from spores from a mushroom of a strain.

(59) Outbreeding: Mating among unrelated or distantly related individuals.

(60) Parent: An immediate progenitor of an individual; a parent strain is a heterokaryon, a parent line is a homokaryon; a heterokaryon may be the parent of an F1 heterokaryon via an intermediate parent line.

(61) Pedigree-assisted breeding: The use of genealogical information to identify desirable combinations of lines in controlled mating programs.

(62) Phenotype: Observable characteristics of a strain or line as expressed and manifested in an environment.

(63) Plasmogamy: Establishment, via anastomosis, of cytoplasmic continuity leading to the formation of a sexual heterokaryon.

(64) Progenitor: Ancestor, including parent (the direct progenitor).

(65) Selfing: Mating among sibling lines; also intramixis.

(66) Somatic: Of the vegetative mycelium.

(67) Spawn: A mushroom culture, typically a pure culture of a heterokaryon, typically on a sterile substrate which is friable and dispersible particulate matter, in some instances cereal grain; commercial inoculum for compost; reference to spawn includes reference to the culture on a substrate.

(68) Spore: Part of the mushroom, the reproductive propagule.

(69) Stem: Stipe; part of the mushroom, the cap-supporting structure.

(70) Sterile Growth Media: Nutrient media, sterilized by autoclaving or other methods, that support the growth of the organism; examples include agar-based solid nutrient media such as Potato Dextrose Agar (PDA), nutrient broth, and many other materials.

(71) Stipe: see stem.

(72) Strain: A heterokaryon with defined characteristics or a specific identity or ancestry; equivalent to a variety.

(73) Tissue culture: A de-differentiated vegetative mycelium obtained from a differentiated tissue of the mushroom.

(74) Trait conversion: Selective introduction of the genetic determinants of one (a single-locus conversion) or more desirable traits into the genetic background of an initial strain while retaining most of the genetic background of the initial strain. See Introgressive trait conversion and Transformation.

(75) Transformation: A process by which the genetic material carried by an individual cell is altered by the incorporation of foreign (exogenous) DNA into its genome; a method of obtaining a trait conversion including a single-locus conversion.

(76) Virus-breaking: Using multiple incompatible strains, i.e. strains exhibiting heterokaryon incompatibility, successively in a program of planned strain rotation within a mushroom production facility to reduce the transmission of virus from on-site virus reservoirs into newly planted crops.

(77) Yield: The net fresh weight of the harvest crop, normally expressed in pounds per square foot.

(78) Yield pattern: The distribution of yield within each flush and among all flushes; influences size, quality, picking costs, and relative disease pressure on the crop and product.

(79) With respect to the definition of homokaryon above, it is noted that homokaryons and homoallelic lines are subject to technical and practical considerations: A homokaryon in classical terms is a haploid culture which is axiomatically entirely homoallelic. In practical terms, for fungal strain development purposes, the definition is broadened somewhat to accommodate both technical limitations and cytological variation, by treating all predominately homoallelic lines as homokaryons. Technical limitations include the fact that genomes contain duplicated DNA regions including repeated elements such as transposons, and may also include large duplications of chromosomal segments due to historical translocation events; such regions may appear not to be homoallelic by most genotyping methods. Two different A. bisporus genomes sequenced by the Joint Genome Institute, a U.S. federal facility, differ in estimated length by 4.4%, and in gene numbers by 8.2%, suggesting a considerable amount of DNA duplication or rearrangement within different strains of the species. No presently available genome of A. bisporus can completely account for the physical arrangement of such elements and translocations, and so the assembled genome sequences of haploid lines may have regions that appear to be heteroallelic using currently available genotyping methods. Cytologically, a homokaryotic offspring will ordinarily be a spore that receives one haploid, postmeiotic nucleus. However, a spore receiving two third-division nuclei from the basidium will be genetically equivalent to a homokaryon. A spore receiving two second-division sister postmeiotic nuclei will be a functional homokaryon even though some distal islands of heteroallelism may be present due to crossovers during meiosis. Also, a meiosis that has an asymmetrical separation of homologues can produce an aneuploid, functionally homokaryotic spore in which an extra chromosome, producing a region of heteroallelism, is present. All of these cultures are highly homoallelic and all function as homokaryons. Technological limitations make it impractical to distinguish among such cultures, and also to rule out DNA segment duplication as an explanation for limited, isolated regions of the genome sequence assembly that appear to be heteroallelic. Therefore, in the present application, the use of the term homoallelic to characterize a line includes entirely or predominately homoallelic lines, and cultures described in this way are functional homokaryons, are putatively homokaryotic, and are all defined as homokaryons in the present application.

(80) Now, with respect to the invention and as noted hereinabove, the present invention relates to cultures of the hybrid Agaricus bisporus strain J11500 and to cultures derived or descended from J11500. Such cultures are used to produce mushrooms and parts of mushrooms. Thus, the present invention further relates to methods of making and using the strain J11500 and Essentially Derived Varieties (EDVs) of the strain J11500.

(81) Hybrid strain J11500 is the product of 6 generations of controlled line matings by Sylvan America, Inc. The original mating was made between line JB 137-s8 and line SWNC. In the sixth generation, line J10102-s69, a descendent of the first hybrid (and of other hybrids produced by Sylvan, Inc.), was mated with line OWNC to produce the novel hybrid strain J11500.

(82) Cultures of strain J11500 produce commercially acceptable and desirable crops of white mushrooms. Table I presents yield data as pounds per square foot, in three independent crop tests with internal replication. As shown in Table I, productivity of J11500 is comparable to and often greater than the productivity of the A15 strain, with total (3-flush) yield averaging 101.3% of the A-15 control and ranging as high as 106.8% under standard growing conditions for A-15. Distribution of the crop over the three-flush harvest period is relatively accelerated, meaning that more of the crop is picked during first flush, when disease pressure is lowest and product quality may be correspondingly higher. In a general t-test on this small data set, first break yield differences between J11500 and A-15 approached statistical significance (p=0.057).

(83) TABLE-US-00001 TABLE I 1.sup.st flush yield 1.sup.st & 2.sup.nd flush yield Total yield Test ID J11500 A-15 J11500 A-15 J11500 A-15 12-108 2.87 2.27 4.50 4.04 5.02 4.70 12-119 2.47 2.15 3.73 3.81 4.34 4.61 12-146 2.57 2.39 3.92 3.71 4.60 4.47 Averages 2.63 2.27 4.05 3.85 4.65 4.59 % gain +16% +5% +1%

(84) Within first flush, yield is also accelerated. Over the four productive days of first flush, the cumulative daily yield data in Table II, reporting averages of the same three tests, shows that the harvest of strain J11500 is accelerated over that of the A-15 control.

(85) TABLE-US-00002 TABLE II Day (after casing): Cumulative daily yield: 14 15 16 17 J11500 yield as a percent of A-15 yield 181% 139% 128% 116%

(86) Timing to harvest is about equivalent to that of commercial strain A15 (both about 13 to 19 days), and sometimes may be slightly faster, which can be economically advantageous. Table III shows that in the same crop tests, on average, strain J11500 began to produce its crop 0.43 days before A-15, and the peak of production in the first flush was 0.24 days earlier for strain J11500.

(87) TABLE-US-00003 TABLE III Days to first pick Peak first flush pick day Test ID J11500 A-15 J11500 A-15 12-108 14.0 15.3 14.7 15.3 12-119 14.0 14.0 14.0 14.3 12-146 14.0 14.0 15.0 14.8 Averages 14.0 14.43 14.56 14.8 Days gained +0.43 +0.24

(88) Cap roundness and relative flesh thickness (i.e., meatiness) are considered to be desirable commercial mushroom traits. J11500 typically produces mushrooms with caps having thicker flesh, and which are subjectively rounder, than those of A15; objectively, the following physical measurement ratios demonstrate the shape differences of J11500 compared to A15.

(89) Cap roundness, expressed as cap height/cap diameter (CH/CD) is an economically important trait reflecting a consumer preference for rounder mushrooms. Measurements were made on samples of 10 first break mushrooms of equivalent maturity from both J11500 and the commercial control A-15. J11500 was rounder (0.68) compared to the control A-15 (0.60), and this difference was significant (t-test, p=9.15E-07).

(90) Similarly, cap meatiness, expressed as flesh thickness/cap diameter (FT/CD) is an economically important trait reflecting a consumer preference for thicker-fleshed mushrooms. Measurements were made on samples of 10 first break mushrooms of equivalent maturity from both J11500 and the commercial control A-15. J11500 was meatier (0.36) compared to the control A-15 (0.33), and this difference was significant (t-test, p=0.0054).

(91) Cross-strainincompatibility can also be a useful commercial mushroom trait. J11500 is incompatible with A-15, a proxy for the U1 derived lineage group. When casing material incorporating inoculum of J11500 is placed over compost colonized with A-15, or conversely when A-15 is placed over J11500, a partial crop failure ensues, demonstrating incompatibility as shown by the yield data in TABLE IV:

(92) TABLE-US-00004 TABLE IV Spawn strain Casing strain Identity First flush yield J11500 J11500 Self 2.47 lbs. A-15 A-15 Self 2.03 lbs. J11500 A-15 Non-self 0.50 lbs. A-15 J11500 Non-self 0.17 lbs.

(93) The incompatibility of J11500 with A-15 is transmitted into spores and thus is inherited by EDVs derived from spores, as shown by the yield data in TABLE V:

(94) TABLE-US-00005 TABLE V Spawn strain Casing strain Identity First flush yield A-15 A-15 Self 1.74 lbs. A-15 J11500-ms2 Non-self 0.58 lbs. A-15 J11500-ms3 Non-self 0.63 lbs. A-15 J11500-ms4 Non-self 0.58 lbs. A-15 J11500-ms5 Non-self 0.44 lbs. A-15 J11500-ms10 Non-self 0.53 lbs.

(95) A test of compatibility of an EDV of strain J11500 (designated J11500-ms2) with the strain J11500 itself was performed and the results are shown in TABLE VI.

(96) TABLE-US-00006 TABLE VI Spawn strain Casing strain Identity First flush yield J11500 J11500 Self 1.95 lbs. J11500 J11500-ms2 Self: EDV 2.69 lbs. J11500 J11500-ms2 Self: EDV 3.13 lbs.

(97) Table VI shows that in test 13-177, the EDV strain designated J11500-ms2 was completely compatible with the initial strain J11500, and in fact demonstrated higher first break yield than strain J11500 as opposed to a partial crop failure that would have indicated incompatibility.

(98) Given that strain J11500 has 4 non-cultivar progenitors and that considerable genetic diversity exists among strains, the genotypic fingerprint of strain J11500 shows numerous differences with that of the U1 lineage group. A unique fingerprint allows strain J11500 (and its Essentially Derived Varieties and descendents) to be unambiguously identified. Agronomically, genetic diversity among cultivated strains is a desirable objective because it is well established that genetic monocultures among agricultural crop species can lead to disastrous failures due to particular disease, pest, or environmental pressures. Any otherwise desirable commercial strain with genetic novelty is therefore valuable. Strain J11500 meets those criteria.

(99) For the purpose of this invention, the whole genomic sequence of strain J11500 and of the cultures of its parent lines and of selected EDVs of J11500 have been obtained by Sylvan America Inc. using the following method. The homokaryotic parent line cultures were grown in sterile broth growth medium after maceration. After 2-4 weeks, hyphal cells were collected by filtration, were frozen at 80 C, and were lyophilized until dry. Cap tissue was obtained from mushrooms produced by cultures of the heterokaryotic J11500 (and EDV) strains, and was frozen and lyophilized. DNA was extracted using a CTAB protocol followed by RNAse treatment and gel purification. A contractor, SeqWright, prepared DNA libraries from the DNA of each culture, and sequenced the libraries using Illumina MiSeq technology. Assemblies of the reads into genomic sequence using the public-domain reference genome sequence of H97 was performed by the contractor. Consequently about 93% to about 95% of the entire genotype of strain J11500 and of three EDVs of strain J11500 are known to Sylvan America, Inc with certainty. The total number of markers distinguishing strain J11500 that are known to the assignee is about 300,000. A brief excerpt of the genotype strain J11500 at numerous sequence-characterized marker loci distributed at intervals along each of the 19 H97 V2.0 reference scaffolds larger than 100 Kb in length is provided in Table VII.

(100) TABLE-US-00007 TABLEVII PositionofSNA Culture: Scaffold [H97V2.0ref.coords.] J10102-s69 OWNC J11500 J11500-ms2 1 99995 CTAC TTGA CTACATTGA CTAC TTGA CTAC TTGA 1 349966 AAGG GGTT AAGGTGGTT AAGG GGTT AAGG GGTT 1 600059 TTTT TTT TTTTTTTT-C TTTT TT[-/A] TTTT TT[-/A] 1 850014 C TTTTC C CCTTTTCAC C TTTTC C C TTTTC C 1 1099971 GTCG CACC GTCGACACC GTCG CACC GTCG CACC 1 1350278 GGAG TCG GGAGAGTCG GGAG TCG GGAG TCG 1 1599956 AATA GCGC AATAAGCGC AATA GCGC AATA GCGC 1 1850032 CGAG AATT CGAGTAATT CGAG AATT CGAG AATT 1 2119049 ACAA CAA ACAATCCAA ACAA CAA ACAA CAA 1 2400243 ACTT ATGA ACTTCATGA ACTT ATGA ACTT ATGA 1 2612870 AATA GAGT AATAGGAGT AATA GAGT AATA GAGT 1 2858975 GCCG TCTT GCCGTTCTT GCCG TCTT GCCG TCTT 1 2804522 GAAG GAC GAAGACGAC GAAG GAC GAAG GAC 1 3047987 AAGG GGGG AAGGGGGGG AAGG GGGG AAGG GGGG 1 3164166 ATAA GGG ATAAGGGGG ATAA GGG ATAA GGG 1 3256057 TATC GTTT TATCTGTTT TATC GTTT TATC GTTT 2 101820 ATTA GAT ATTAAAGAT ATTA GAT ATTA GAT 2 350156 TCGG GGTG TCGGGGGTG TCGG GGTG TCGG GGTG 2 600112 ATGT TACG ATGTATACG ATGT TACG ATGT TACG 2 850338 TGGT CTAA TGGTGCTAA TGGT CTAA TGGT CTAA 2 1099413 CCTG CTCA CCTGACTCA CCTG CTCA CCTG CTCA 2 1349512 CTCA CAGT CTCAGCAGT CTCA CAGT CTCA CAGT 2 1600085 CACA TGCC CACAATGCC CACA TGCC CACA TGCC 2 1901773 ACTC AATT ACTCGAATT ACTC AATT ACTC AATT 2 2150201 GTCG AGGT GTCGTAGGT GTCG AGGT GTCG AGGT 2 2400281 TCAA AC C TCAAAACCC TCAA AC C TCAA AC C 2 2650136 ATAA TCCT ATAATTCCT ATAA TCCT ATAA TCCT 2 2903593 ACTA A GA ACTAAAAGA ACTA A GA ACTA A GA 2 3048019 GTCC CTGC GTCCGCTGC GTCC CTGC GTCC CTGC 3 65650 GGCG TTTT GGCGCTTTT GGCG TTTT GGCG TTTT 3 119281 TTTA ACTC TTTATACTC TTTA ACTC TTTA ACTC 3 249570 GTAT ATGT GTAT ATGT GTAT ATGT GTAT ATGT 3 750000 GTCC GCCA GTCC GCCA GTCC GCCA GTCC GCCA 3 1250000 TTTT CCGG TTTT CCGG TTTT CCGG TTTT CCGG 3 1750000 ACGC TGAC ACGC TGAC ACGC TGAC ACGC TGAC 3 2250000 CGTG CGAT AGTG CGAT CGTG CGAT CGTG CGAT 3 2520748 TAAT CCAC TAATGCCAC TAAT CCAC TAAT CCAC 4 100004 GAGT AT A GAGTGATAA GAGT AT A GAGT AT A 4 340893 AGG GGTA AGGTGGTAT AGG GGTA AGG GGTA 4 598147 GATC ACAG GATCGACAG GATC ACAG GATC ACAG 4 852119 CGAA A TC CGAATATTC CGAA A TC CGAA A TC 4 1100085 GATG CGAA GATGCCGAA GATG CGAA GATG CGAA 4 1350536 CGAA CGG CGAACTCGG CGAA CGG CGAA CGG 4 1599885 GATA TTGC GATACTTGC GATA TTGC GATA TTGC 4 1850288 ATTC GTA ATTCGTGTA ATTC GTA ATTC GTA 4 2100356 TCAG GACC TCAGAGACC TCAG GACC TCAG GACC 4 2284257 TCTG ACTG TCTGGACTG TCTG ACTG TCTG ACTG 5 100211 TCCT GAAT TCCTTGAAT TCCT GAAT TCCT GAAT 5 350872 GGCG GCCC GGCGTGCCC GGCG GCCC GGCG GCCC 5 599922 CGTC TTCA CGTCATTCA CGTC TTCA CGTC TTCA 5 851262 TAAT TCT TAATTCTCT TAAT TCT TAAT TCT 5 1099776 ACAT GACA ACATTGACA ACAT GACA ACAT GACA 5 1352539 TTGT TCC TTGTGATCC TTGT TCC TTGT TCC 5 1599904 AACT CCTT AACTTCCTT AACT CCTT AACT CCTT 5 1851458 AAAT TCC AAATAATCC AAAT TCC AAAT TCC 5 2100025 CCCT AGTC CCCTTAGTC CCCT AGTC CCCT AGTC 5 2278878 GGTC AAAA GGTCGAAAA GGTC AAAA GGTC AAAA 6 106294 GCCA CTC GCCATCTCG GCCA CTC GCCA CTC 6 350337 CATT GGTT CATTTGGTT CATT GGTT CATT GGTT 6 600047 GGAG ATTT GGAGCATTT GGAG ATTT GGAG ATTT 6 849990 AGTT AGGA AGTTCAGGA AGTT AGGA AGTT AGGA 6 1098535 CAAA ATTG CAAAGATTG CAAA ATTG CAAA ATTG 6 1349453 TGTC TAG TGTCGGTAG TGTC TAG TGTC TAG 6 1600000 AAAC TGGA AAAC TGGA AAAC TGGA AAAC TGGA 6 1676645 AACC GATT AACCGGATT AACC GATT AACC GATT 6 2000087 GATT TGCG GATTTTGCG GATT TGCG GATT TGCG 6 2252662 GGGT GGTA GGGTTGGTA GGGT GGTA GGGT GGTA 7 100284 GAAA TCAG GAAATTCAG GAAA TCAG GAAA TCAG 7 350044 ATAT CTTT ATATTCTTT ATAT CTTT ATAT CTTT 7 600111 CAAT ATTA CAATTATTA CAAT ATTA CAAT ATTA 7 850516 TGAC CATA TGACGCATA TGAC CATA TGAC CATA 7 1100248 TCAC GAAG TCACGGAAG TCAC GAAG TCAC GAAG 7 1350089 CTTT CCCC CTTTTCCCC CTTT CCCC CTTT CCCC 7 1605047 ATAC TG C ATACTTGGC ATAC TG C CTAC TG C 7 1850000 GAGA ACT GAGA ACT GAGA ACT GAGA ACT 7 1898793 TCCG AT A TCCGCATAA TCCG AT A TCCG AT A 7 1991505 TCTA GTT TCTACGGTT TCTA GTT TCTA GTT 8 350000 ATTG CGCG ATTG CGCG ATTG CGCG ATTG CGCG 8 600000 CATT ACGG CATT ACGG CATT ACGG CATT ACGG 8 1100000 CATA GATC CATA GATC CATA GATC CATA GATC 8 1350000 AGCT AACA AGCT AACA AGCT AACA AGCT AACA 8 1600100 CTGA CCCT CTGA CCCT CTGA CCCT CTGA CCCT 9 100105 CTCA CCGA CTCAACCGA CTCA CCGA CTCA CCGA 9 352455 AGTC CCA AGTCCTCCA AGTC CCA AGTC CCA 9 599950 TGGT TCCC TGGTATCCC TGGT TCCC TGGT TCCC 9 1010845 GGGT GTGA GGGTGGTGA GGGT GTGA GGGT GTGA 9 1244202 GATG AGAT GATGAAGAT GATG AGAT GATG AGAT 9 1504476 TACT TACC TACTGTACC TACT TACC TACT TACC 9 1656962 TATC ACTG TATCTACTG TATC ACTG TATC ACTG 10 100438 AATT ATTT AATTAATTT AATT ATTT AATT ATTT 10 350030 GCGG TCAA GCGGCTCAA GCGG TCAA GCGG TCAA 10 600032 TTAC CTGG TTACACTGG TTAC CTGG TTAC CTGG 10 850000 TCGG CGGA TCGG CGGA TCGG CGGA TCGG CGGA 10 860249 CCGC AAATT CCGCAAATT CCGC AAATT CCGC AAATT 10 1109960 AGGA ATGA AGGAAATGA AGGA ATGA AGGA ATGA 10 1303902 TGAT TACT TGATTTACT TGAT TACT TGAT TACT 10 1490452 AATC GATG AATCAGATG AATC GATG AATC GATG 11 100000 TATT TTAG TATT TTAG TATT TTAG TATT TTAG 11 350000 GTCA CAAG GTCA CAAG GTCA CAAG GTCA CAAG 11 600000 ATGG CGCG ATGG CGCG ATGG CGCG ATGG CGCG 11 850000 CTTC CCAT CTTC CCAT CTTC CCAT CTTC CCAT 11 1100000 TTAC GTTG TTAC GTTG TTAC GTTG TTAC GTTG 11 124000 AGCC AGTA AGCC AGTA AGCC AGTA AGCC AGTA 12 100000 CCTT TAGT CCTT TAGT CCTT TAGT CCTT TAGT 12 1000000 CGAG AGGA CGAG AGGA CGAG AGGA CGAG AGGA 13 100697 ACGT TTTA ACGTCTTTA ACGT TTTA ACGT TTTA 13 370521 TTTG GTCA TTTGAGTCA TTTG GTCA TTTG GTCA 13 604345 CTTC GCAT CTTCAGCAT CTTC GCAT CTTC GCAT 13 850249 GG T GT A GGCTAGTAA GG T GT A GG T GT A 14 113109 AGGG AATA AGGGAAATA AGGG AATA AGGG AATA 14 372086 CGAT C TT CGATCCCTT CGAT C TT CGAT C TT 14 725684 ATGA TT G ATGAGTTCG ATGA TT G ATGA TT G 15 150013 GTGG CCGT GTGGCCCGT GTGG CCGT GTGG CCGT 15 449866 GAAT TCGG GAATTTCGG GAAT TCGG GAAT TCGG 16 208609 CACA GCAC CACATGCAC CACA GCAC CACA GCAC 16 400000 CCTC GATT CCTC GATT CCTC GATT CCTC GATT 17 120000 TATT TTCA TATT TTCA TATT TTCA TATT TTCA 17 338415 TGAG AGCC TGAGAAGCC TGAG AGCC TGAG AGCC 17 449833 ATCA AC A ATCAGACAA ATCA AC A ATCA AC A 18 101884 ATTA GGAC ATTACGGAC ATTA GGAC ATTA GGAC 19 98377 GCTA TGGG GCTATTGGG GCTA TGGG GCTA TGGG

(101) Table VII presents a fingerprint excerpted from the SNP (Single Nucleotide Polymorphism) marker genotype of the entire genome sequences of line J10102-s69, of line OWNC, of the F1 hybrid J11500 strain obtained from the mating of lines J10102-s69 and OWNC, and of the J11500-ms2 EDV of strain J11500. The IUPAC nucleotide and ambiguity codes are used to represent the observed 9-base DNA marker sequences reported above, each of which represents a genotypic marker locus. The identity of each marker locus is specified by the scaffold and SNP position information derived from the H97 V2.0 reference genome sequence published by the U.S. Department of Energy Joint Genome Institute (Morin et al. 2012). It is evident that a composite relationship of the heteroallelic genotype of strain J11500 exists with respect to the homoallelic genotypes of its two parental lines, namely line J10102-s69 and line OWNC. It is further evident that the heterokaryon genotype of the example EDV J11500-ms2 matches that of its initial strain, J11500.

(102) A brief description of the genotype of strain J11500 at further six unlinked marker loci is provided below. Because the J11500 heterokaryon incorporates two sets of chromosomes, there are two allelic copies (two characters or elements of the genotype) at each marker locus. The brief genotype excerpt provided below therefore consists of 12 characters or elements. The brief genotype was prepared by the assignee of record using targeted Polymerase Chain Reactions to amplify genomic regions bracketing the defined markers from each of the culture DNAs. Any suitable PCR primers that bracket the defined marker regions may be used for this purpose; methods of designing suitable primers are well known in the art. The amplified PCR product DNA was sequenced by a contractor, Eurofins, using methods of their choice, and the genotypes were determined by direct inspection of these sequences in comparison to Sylvan America's database of reference marker/allele sequences.

(103) Description of the p1n150-G3-2 Marker:

(104) The 5 end of this marker segment begins at position 1 with the first T in the sequence TCCCAAGT, corresponding to H97 JGI V2.0 Scaffold 1 position 868615 (Morin et al. 2012) and extending in a reverse orientation (relative to the scaffold orientation) for ca. 600 nt in most alleles; an insertion in the DNA of allele 1T has produced a longer segment. At present, 9 alleles incorporating at least 30 polymorphic positions have been documented from diverse strains in Sylvan America's breeding collection.

(105) Alleles present in the J10102-s69 and J11500 pedigree over three generations are alleles 1T, 2, 3, 4, and 9, characterized as follows (using the format: nucleotide base character @ alignment position, based on alignment of alleles 2, 3, and 4, and the alignable portions of allele 1T):

(106) Allele 1T: C @ 193; insertion of Abr1 transposon of 320 nt @ 206^207; T @ 327; C @ 374; G @ 378; G @ 422; C @ 431; G @ 472; etc.

(107) Allele 2: no Abr1 insertion; C @ 193; C @ 327, C @ 374; C @ 378; G @ 422; T @ 431; G @ 472; etc.

(108) Allele 3: no Abr1 insertion; C @ 193; T @ 327, G @ 374; C @ 378; G @ 422; T @ 431; A @ 472; etc.

(109) Allele 4: no Abr1 insertion; C @ 193; T @ 327, C @ 374; C @ 378; A @ 422; T @ 431; G @ 472; etc.

(110) Allele 9: no Abr1 insertion; G @ 193; C @ 327, C @ 374; C @ 378; G @ 422; T @ 431; G @ 472; etc.

(111) Because of linkage to the MAT locus, which is obligately heteroallelic in fertile heterokaryons, genotypes of all known and expected heterokaryons at p1n150-G3-2 are also heteroallelic.

(112) The J10102 heterokaryon has an 1/2 heteroallelic genotype.

(113) The U1 heterokaryon has an 1/2 heteroallelic genotype.

(114) Off-White heterokaryons such as Somycel 76 have a 1/3 heteroallelic genotype.

(115) Smooth-White heterokaryons such as Somycel 53 have a 2/3 heteroallelic genotype.

(116) The J9277 heterokaryon has a 1/4 heteroallelic genotype.

(117) The genotype of the J11500 heterokaryon at the p1n150-G3-2 marker locus is 1/2 (heteroallelic), designating the presence of alleles 1 and 2. Allele 1 was contributed by the OWNC line. Allele 2 was transmitted from the J10102 heterokaryon via the J10102-s69 homokaryon. The 1/2 genotype distinguishes J11500 from many other heterokaryons including from all of its own grandparents, although not from the U1 strain family.

(118) Description of the ITS (=ITS 1+2 Region) Marker:

(119) The ITS segment is part of the nuclear rDNA region, which is a cassette that is tandemly repeated up to an estimated 100 times in the haploid genome of A. bisporus. Therefore there is no single precise placement of this sequence in the assembled H97 genome, and in fact it is difficult or impossible to precisely assemble the sequence over all of the tandem repeats. Three cassette copies were included on scaffold 10 of the H97 JGI V2.0 assembly, beginning at position 1612110; a partial copy is also assembled into scaffold 29 (Morin et al. 2012). The 5 end of this marker segment begins at position 1 with the first G in the sequence GGAAGGAT, and extending in a forward orientation (relative to the scaffold orientation) for ca. 703-704 nt in most alleles. At present, more than 9 alleles incorporating at least 11 polymorphic positions have been documented from diverse strains in Sylvan's breeding collection.

(120) Alleles present in the J10102-s69 and J11500 immediate pedigree are alleles I1, I2, and I4, characterized as follows (using the format: nucleotide base character @ alignment position, based on alignment of 9 alleles).

(121) Allele I1: C @ 52; T @ 461; T @ 522; T @ 563; etc.

(122) Allele I2: T @ 52; T @ 461; T @ 522; T @ 563; etc.

(123) Allele I4: C @ 52; A @ 461; C @ 522; C @ 563; etc.

(124) The J10102 heterokaryon has an I1/I4 heteroallelic genotype.

(125) The U1 heterokaryon has an I1/I2 heteroallelic genotype.

(126) The genotype of the J11500 heterokaryon at the ITS marker locus is I1/I4 (heteroallelic), designating the presence of alleles I1 and I4. Allele I1 was contributed by the OWNC line. Allele I4 was transmitted from the J10102 heterokaryon via the J10102-s69 homokaryon. This distinguishes J11500 from the U1 strain family, which has an I1/I2 genotype.

(127) Description of the MFPC-1-ELF Marker:

(128) The 5 end of this marker segment begins at position 1 with the first G in the sequence GGGAGGGT, corresponding to H97 JGI V2.0 Scaffold 8 position 829770 (Morin et al. 2012) and extending in a forward orientation (relative to the scaffold orientation) for ca. 860 nt in most alleles. At present, at least 7 alleles incorporating at least 40 polymorphic positions have been documented from diverse strains in Sylvan's breeding collection.

(129) Alleles present in the J10102-s69 and J11500 immediate pedigree, are alleles E1, E2, and E8, characterized as follows (using the format: nucleotide base character @ alignment position, based on alignment of 8 alleles).

(130) Allele E1: A @ 77; A @ 232; A @ 309; T @ 334; A @ 390; A @ 400; T @ 446, A @ 481; etc.

(131) Allele E2: G @ 77; A @ 232; G @ 309; T @ 334; G @ 390; G @ 400; C @ 446, G @ 481; etc.

(132) Allele E8: A @ 77; G @ 232; G @ 309; A @ 334; A @ 390; A @ 400; C @ 446, G @ 481; etc.

(133) The J10102 heterokaryon has an E1/E8 heteroallelic genotype.

(134) The U1 heterokaryon has an E1/E2 heteroallelic genotype.

(135) The genotype of the J11500 heterokaryon at the MFPC-1-ELF marker locus is E1/E1, designating the presence of two copies of alleles E1. One copy of allele E1 was contributed by the OWNC line; a second copy of allele E1 was transmitted from the J10102 heterokaryon via the J10102-s69 homokaryon. This homoallelic genotype distinguishes J11500 from the predominant U1-type of commercial cultivar, which has an E1/E2 genotype.

(136) Description of the AN marker:

(137) The 5 end of this marker segment begins at position 1 with the first G in the sequence GGGTTTGT, corresponding to H97 JGI V2.0 Scaffold 9 position 1701712 (Morin et al. 2012) and extending in a forward orientation (relative to the scaffold orientation) for ca. 1660 nt (in the H97 genome) to 1700 nt (in the alignment space) in known alleles; several insertions/deletions have created length polymorphisms which, in addition to point mutations of individual nucleotides, characterize the alleles. At present, 5 alleles incorporating more than 70 polymorphic positions have been documented from diverse strains in Sylvan's breeding collection.

(138) Alleles present in the J10102-s69 and J11500 immediate pedigree are alleles N1, N2 and N5, characterized in part as follows (using the format: nucleotide base character @ alignment position, based on alignment of alleles N1 through N5):

(139) Allele N1: G @ 640; [deletion] @ 844-846; T @ 882; A @ 994, etc.

(140) Allele N2: A @ 640; [deletion] @ 844-846; T @ 882; A @ 994, etc.

(141) Allele N5: A @ 640; ACG @ 844-846; C @ 882; G @ 994, etc.

(142) The J10102 heterokaryon has an N1/N5 heteroallelic genotype.

(143) The U1 heterokaryon has an N1/N2 heteroallelic genotype.

(144) The genotype of the J11500 heterokaryon at the AN marker locus is N1/N5 (heteroallelic), designating the presence of alleles N1 and N5. Allele N1 was contributed by the OWNC line. Allele N5 was transmitted from the J10102 heterokaryon via the J10102-s69 homokaryon.

(145) The N1/N5 genotype at the AN marker locus distinguishes J11500 from commercial strains U1 and A-15, which have an N1/N2 genotype. This element of the genotype fingerprint can also distinguish J11500 from among many other strains.

(146) Description of the AS Marker:

(147) The 5 end of this marker segment begins at position 1 with the first G in the sequence GG(T/N)GTGAT, corresponding to H97 JGI V2.0 Scaffold 4 position 752867 (Morin et al. 2012) and extending in a forward orientation (relative to the scaffold orientation) for ca. 1620 nt (in the H97 genome) to 1693 nt (in the alignment space) in known alleles; several insertions/deletions have created length polymorphisms which, in addition to point mutations of individual nucleotides, characterize the alleles. At present, 7 alleles incorporating more than 80 polymorphic positions have been documented from diverse strains in Sylvan's breeding collection.

(148) Alleles present in the J10102-s69 and J11500 immediate pedigree are alleles SC and SD, characterized in part as follows (using the format: nucleotide base character @ alignment position, based on alignment of alleles SA through SG):

(149) Allele SC: T @ 28; GATATC @ 258-263; G @ 275; [insertion]+VTTTCTCAGC+[insertion] @ 309-349; C @ 404, etc.

(150) Allele SD: C @ 28; [deletion] @ 258-263; T @ 275; [deletion] @ 309-349; T @ 404, etc.

(151) The J10102 heterokaryon has an SC/SD heteroallelic genotype.

(152) The U1 heterokaryon has an SC/SD heteroallelic genotype.

(153) The genotype of the J11500 heterokaryon at the AS marker locus is SC/SD (heteroallelic), designating the presence of alleles SC and SD. Allele SD was contributed by the OWNC line. Allele SC was transmitted from the J10102 heterokaryon via the J10102-s69 homokaryon.

(154) The SC/SD genotype at the AS marker locus is also shared by commercial strains U1 and A-15. While this element of the genotype fingerprint distinguished J11500 from among many other strains, it does not distinguish J11500 from the U1 strain family.

(155) Description of the FF Marker:

(156) The 5 end of this marker segment begins at position 1 with the first T in the sequence TTCGGGTG, corresponding to H97 JGI V2.0 Scaffold 12 position 281674 (Morin et al. 2012) and extending in a forward orientation (relative to the scaffold orientation) for ca. 570 nt in most alleles. At present, 7 alleles incorporating at least 20 polymorphic positions have been documented from diverse strains in Sylvan's breeding collection.

(157) Alleles present in the J10102-s69 and J11500 immediate pedigree are Alleles FF1 and FF2, characterized as follows (using the format: nucleotide base character @ alignment position, based on alignment of alleles 1 and 2):

(158) Allele FF1: CCG @ 48-50

(159) Allele FF2: TTC @ 48-50

(160) The J10102 heterokaryon has an FF1/FF2 heteroallelic genotype.

(161) The U1 heterokaryon has an FF1/FF2 heteroallelic genotype.

(162) The genotype of the J11500 heterokaryon at the FF marker locus is FF1/FF1 (homoallelic), designating the presence of two copies of allele FF-1, contributed by both the OWNC line and the J10102-s69 homokaryon. This distinguishes J11500 from the predominant U1-type of commercial cultivar, which has an FF1/FF2 genotype. This element of the genotype fingerprint can also distinguish J11500 from among many other strains.

(163) A deposit of a culture of an example of an EDV, namely strain J11500-ms2, obtained from hybrid strain J11500, as disclosed herein, has also been made with the Agricultural Research Services Culture Collection (NRRL) 1815 North University Street, Peoria, Ill. 61604 USA. The date of deposit was Jan. 15, 2014. The culture deposited was taken from the same culture maintained by Sylvan America, Inc., Kittanning, Pa., the assignee of record, since prior to the filing date of this application. All restrictions upon the deposit have been removed, and the deposit is intended to meet all deposit requirements of the U.S. Patent and Trademark Office, including 37 C.F.R. Sec. 1.801-1.809, and all deposit requirements under the Budapest Treaty. The NRRL Accession No. is 50896. The deposit 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 as necessary during that period. The culture will be irrevocably and without restriction or condition released to the public upon filing of a priority application or upon the issuance of a patent according to the patent laws.

(164) Further, the two parent lines of J11500 have also been deposited. Specifically, a deposit of a culture of the Agaricus bisporus homokaryotic line J10102-s69, as disclosed herein, has been made with the Agricultural Research Services Culture Collection (NRRL) 1815 North University Street, Peoria, Ill. 61604 USA. The date of deposit was Jan. 15, 2014. The culture deposited was taken from the same culture maintained by Sylvan America, Inc., Kittanning, Pa., the assignee of record, since prior to the filing date of this application. All restrictions upon the deposit have been removed, and the deposit is intended to meet all deposit requirements of the U.S. Patent and Trademark Office, including 37 C.F.R. Sec. 1.801-1.809, and all deposit requirements under the Budapest Treaty. The NRRL Accession No. is 50893. The deposit 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 as necessary during that period. The culture will be irrevocably and without restriction or condition released to the public upon filing of a priority application or upon the issuance of a patent according to the patent laws.

(165) In addition, a deposit of a culture of the Agaricus bisporus homokaryotic line OWNC, as disclosed herein, has been made with the Agricultural Research Services Culture Collection (NRRL) 1815 North University Street, Peoria, Ill. 61604 USA. The date of deposit was Jan. 15, 2014. The culture deposited was taken from the same culture maintained by Sylvan America, Inc., Kittanning, Pa., the assignee of record, since prior to the filing date of this application. All restrictions upon the deposit have been removed, and the deposit is intended to meet all deposit requirements of the U.S. Patent and Trademark Office, including 37 C.F.R. Sec. 1.801-1.809, and all deposit requirements under the Budapest Treaty. The NRRL Accession No. is 50894. The deposit 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 as necessary during that period. The culture will be irrevocably and without restriction or condition released to the public upon filing of a priority application or upon the issuance of a patent according to the patent laws.

(166) One use of the culture of strain J11500 is the production of crops of edible mushrooms for sale. Another use is for the improvement of facility hygiene via strain rotation and a virus-breaking effect. A third use is to incorporate the genetic material of strain J11500 into offspring and derived or descended cultures including dormant and germinating spores and protoplasts. Additional uses also exist as noted above.

(167) Hybridization of Agaricus bisporus cultures of the invention may be accomplished by allowing two different cultures, one of which is a genetic line present in a spore of J11500, to grow together in close proximity, preferably on sterile media, until anastomosis (i.e., hyphal or cell fusion) occurs. In a successful mating, the resultant fusion culture is a first-generation outbred hybrid culture incorporating a genetic line present in a mushroom spore which is one part of one embodiment of the present invention. Protoplasts derived from basidia or other parts of the organism are another part of the J11500 mushroom that may be used to transmit genetic material of J11500 into new cultures.

(168) Methods for obtaining, manipulating, and mating cultures of the present invention, for producing offspring, inoculum, products, and crops of the current invention, for using a strain rotation program to improve mushroom farm hygiene, and for obtaining the genotypic fingerprint of mushroom cultures, are described hereinabove and are also well known to practitioners of the art.

(169) In order to demonstrate practice of the invention, a subculture of strain J11500 was propagated as described above to produce spawn and casing inocula, which were used to produce crops of white mushrooms under standard commercial cultivation practices as described herein above (see Background of Invention section). Commercial culture inocula including mushroom spawn and casing inoculum were prepared using commercial large-scale microbiological production methods, namely, by aseptically introducing inoculum of a pure culture of strain J11500 into from one to about 2,000 liters of sterilized growth media under sterile conditions, and were disbursed into sterile packaging for test purposes. The mushroom spawn was mixed with pasteurized compost and incubated for 13 to 18 days. A non-nutritive peat-based casing layer was placed over the compost as previously described and a casing inoculum was incorporated into the casing layer. Under controlled environmental conditions, the first mushrooms reached the correct stage of development in a further 14 days. The mushrooms were picked over a 3 to 4 day period. Three flushes of mushrooms were harvested before each test was concluded.

(170) The mushrooms produced by strain J11500 have a white pileus color. As the Royal Horticultural Society (RHS) color charts do not provide a reference standard for the color white, direct measurements of color of the strain J11500 mushroom cap have been made using a Minolta Chromameter and the L-a-b color space system. One measurement was made on the caps of each of ten first break mushrooms grown in a testing facility. The mean values, plus or minus the standard error, for the measured L, a, and b color components were as follows: L=89.580.11; a=1.210.015; b=8.120.088. Colors within or substantially coinciding with color space described by these three parameter distributions are called white according to standard and accepted practices of the commercial mushroom industry.

(171) Strain J10102 is a heterokaryotic strain obtained in Sylvan America, Inc.'s strain development program. It did not have the combination of characters needed to be successful commercially; however its performance and physical characteristics approached those criteria, and the strain was assessed as having some unknown potential for further development and improvement. Consequently, J10102 was used as a parent in 165 matings to several diverse lines of A. bisporus that, it was believed, might have had some useful potential in mating combinations. Individual outcomes were unpredictable and variable; it was hoped that the experiment might produce a successful result but the overall likelihood of that was considered to be low. Of the 165 novel hybrids obtained, only two were of potential commercial interest, and only one, J11500, consistently met the target criteria for a successful commercial strain. It was later determined in the course of testing that strain J11500 had other beneficial attributes as well.

(172) Essentially Derived Varieties of strain J11500 were obtained from single spores, multiple spore mixtures, and from tissue and somatic selections, as described hereinabove. Spores of strain J11500 were obtained and were germinated and used to produce heterokaryotic and homokaryotic offspring as described hereinabove. Homokaryotic offspring lines were used to make matings to other lines, and further hybrids were obtained from these matings. Spawn and casing inoculum of J11500 and A-15 were used in self/self and self/non-self combinations in test crops to confirm the incompatibility of the two strains, a prerequisite for use in virus-breaking strategies, all as described hereinabove.

(173) Although the invention has been described in terms of particular embodiments in this application, one of ordinary skill in the art, in light of the teachings herein, can generate additional embodiments and modifications without departing from the spirit of, or exceeding the scope of, the claimed invention. Accordingly, it is understood that the descriptions herein are proffered only to facilitate comprehension of the invention and should not be construed to limit the scope thereof.