MUSHROOM VARIETY 'RO10425'
20260060156 ยท 2026-02-26
Assignee
Inventors
Cpc classification
International classification
Abstract
A new and unique brown variety of the mushroom Agaricus bisporus Imbach was produced by crossing the progeny of brown mushrooms and off-white hybrid mushrooms to bridging cross strains. The resultant hybrid, named RO10425, exhibits distinct and stable characteristics. Hybrid RO10425 exhibits a lighter cap color, thicker cap flesh, and rounder cap, especially when compared to available brown mushroom varieties. RO10425 also exhibits fast mushroom development and high yield.
Claims
1. A new and distinct variety of Agaricus bisporus Imbach mushroom named RO10425, as shown and described herein.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0022] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawings will be provided by the office upon request and payment of the necessary fec.
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DETAILED BOTANICAL DESCRIPTION
[0028] In mushroom breeding, mycelial (=vegetative) cultures of two compatible progenitors (typically these are haploid homokaryotic strains called homokaryons) must come into physical contact so that one or more fusion zones can occur between the progenitors. Within those fusion zones nuclei from the two progenitors become associated. In Agaricus bisporus, a novel, hybrid mycelium ultimately containing two compatible haploid nuclear types (one from each of the two progenitors) plus one mitochondrial type (from either one of the progenitors) emerges. This novel hybrid mycelium can be isolated and propagated to provide the new hybrid culture, which can be further subdivided and propagated for commercial or other purposes.
[0029] It should be recognized that the mushrooms of the present disclosure are hybrids (equivalent to crosses), as they are formed by the hybridization of at least two independently derived strains, since they were formed by hybridizing the wild strain AA-0096 with a second, commercialized strain of A. bisporus (with the second strain, in some embodiments, being itself a cross between at least two other identified strains of A. bisporus ). The term hybrid mushroom may be used herein to represent the product of a cross between a wild A. bisporus strain and a commercial strain, often including a cross between a brown mushroom strain and an off-white hybrid variety. Differences in terminology (and in the specific techniques used alongside the terminology) are merely indicative of the variety of terminology and techniques used in the mushroom production field.
[0030] After parental homokaryotic spores germinated into mycelia, single spore isolates (SSIs) were selected, preferentially those exhibiting slow mycelial growth. The slowly growing mycelia of SSIs usually contain one nuclear type, are thus homokaryotic, and are suitable for crossing, and thus for breeding. Slow growers are denoted as sg. Some homokaryons were obtained through protoplasting; this technique involves treating the vegetative mycelia of the heterokaryons to damage the fungal cell wall, then selecting colonies of protoclones that are each derived from a single, regenerated protoplast. All crosses were initially cultivated on a small scale, and spores were collected for the parents of the crosses to produce the hybrids described hercin.
[0031] One useful feature in the breeding programs for these novel mushroom varieties is the use of a bridging cross strain, which is a hybrid variety produced from crossing a commercial brown variety with a commercial white variety. The particular bridging cross strain of note in these breeding programs is known as the 4x29 strain. Strain 4x29 has been previously deposited under the provisions of the Budapest Treaty with the American Type Culture Collection, Rockville, Md., USA, ATCC accession No. PTA-6877 (deposited on Jul. 20, 2005). Strain 4x29 is the product of crossing a commercial brown strain with a commercial white strain. The 29 of this strain is a unique homokaryon collected from an off-white hybrid strain.
[0032] The varieties, mushrooms, strains, and cultures disclosed herein are understood to comprise parts of mushrooms, such as the fruiting body, cap, stipe, basidium, spore, mycelium, mixed or unmixed spawn, inoculated substrate, protoplast, gill, cell, or product produced from the mushroom. In some embodiments, the mushroom part from the hybrid mushroom produced by any method disclosed herein is a spore. In some embodiments, the mushroom part produced from a mushroom strain disclosed herein is a spore. In some embodiments, RO14176 is a mushroom part produced from the hybrid mushroom of RO10425. In some embodiments, the mushroom part produced from the hybrid mushroom of RO10425 is RO14176.
Trait Determinations
[0033] In the descriptions and tables that follow, numerous traits are calculated/measured as follows in Table 1. Visual depictions of the measurements used for calculating Cap Apex Flesh Thickness are shown in
[0034] Mushrooms were harvested at Stage 3 as described by Hammond & Nichols (Hammond, J. B. W., & Nichols, R. (1976). Carbohydrate metabolism in Agaricus bisporus (Lange) Sing.: changes in soluble carbohydrates during growth of mycelium and sporophore. Microbiology, 93(2), 309-320), when cap diameters were approx. 3-5 cm with the veil still intact. To reduce confounding effects introduced by different trials, developmental timing and yield traits were compared only from trials where all strains being compared were present together.
[0035] Images of mushroom caps and longitudinal cross-sections were captured using a Canon Rebel SL1 camera (Canon, Inc., Toyko, Japan) in a custom-built photo box with standardized lighting conditions. The camera was calibrated for color consistency using a ColorChecker gray balance card (Calibrite LLC, Wilmington, DE). L*, a* and b* values were captured across the surface of each mushroom cap and cross-section using ImageJ (Schneider, C. A., Rasband, W. S., & Eliceiri, K. W. (2012). NIH Image to ImageJ: 25 years of image analysis. Nature Methods, 9(7), 671-675). Images captured in 2024 and 2025 were used for analysis. Extreme outliers were excluded from the data, defined as those falling 3 times the interquartile range (IQR) above the 75th percentile for that trait or more than 3 times the IQR below the 25th percentile for that trait.
[0036] All statistical analyses were carried out in the R programming language (R Core Team, (2013). R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. www[dot]R-project[dot]org/). Levene's test was performed using the car package (Fox, J., & Weisberg, S. (2018). An R companion to applied regression. SAGE Publications), Shapiro-Wilk tests and Tukey HSD were performed using the stats package (R Core Team (2013)), and standard ANOVAs, Kruskal-Wallis tests and Dunn's tests were performed using the rstatix package (Kassambara, A (2023). rstatix: Pipe-Friendly Framework for Basic Statistical Tests. R package version 0.7.2, rpkgs[dot]datanovia[dot]com/rstatix/).
TABLE-US-00001 TABLE 1 Trait Tool Measurement Scale Timing Cap Color Darkness visually 0 to 5 (from white to dark brown), 1st break**, assessed tray-level measurement 2nd break** Cap Color a* Value image 128 to +127 (from pure green to pure 1st break**, (Green-Red) analysis red), individual mushroom-level 2nd break** measurement Cap Color b* Value image 128 to +127 (from pure blue to pure 1st break**, (Blue-Yellow) analysis yellow), individual mushroom-level 2nd break** measurement Cap Color L* Value image 0 to 100 (from pure black to pure white), 1st break**, (Dark-Light) analysis individual mushroom-level measurement 2nd break** Cap Diameter image millimeters (mm), individual mushroom- 1st break**, analysis level measurement 2nd break** Cap Firmness manual 1 to 5 (from very soft to very firm), tray- 1st break**, squeeze test level measurement 2nd break** Cap Apex Flesh calculated distance from top of gills to top of cap of 1st break**, Thickness ratio and mushroom, relative to cap height, 2nd break** image individual mushroom-level measurement analysis Cap Edge Flesh calculated sum of distances from outer edge of each 1st break**, Thickness ratio and gill to outer edges of cap of mushroom, 2nd break** image relative to cap diameter, individual analysis mushroom-level measurement Cap Flesh Thickness hand- millimeters (mm), distance from top of 1st break**, measurement gill-stipe connection (at center of stipe) to 2nd break** top of cap, individual mushroom-level measurement Cap Height image millimeters (mm), distance from base of 1st break**, analysis cap to top of cap, individual mushroom- 2nd break** level measurement Cap Roundness image Unitless ratio (0-1), 1st break**, (Calculated) analysis calculated as 4 * CS.sub.A/( * CS.sub.D.sup.2), where 2nd break** CS.sub.A = area of the cross-section, and CS.sub.D = diameter of the cross-section; assessed with image analysis, individual mushroom-level measurement Cap Roundness visually 1 to 5 (from very flat to spherical), 1st break**, (Visual) assessed tray-level measurement 2nd break** Compatibility calculated pounds per square foot (lb/ft.sup.2) and 1st break** ratio kilograms per square meter (kg/m.sup.2), weight of 1st break mushrooms yielded relative to area of tray, tray-level measurement Compost Colonization visually 0 to 5 (from no colonization to robust before 1st break assessed colonization), tray-level measurement (10 days after spawning) Days to 1st Break visually number of days elapsed after casing at 1st break** assessed which at least half of tray is of harvestable maturity for 1st break, tray-level measurement Days to 2nd Break visually number of days elapsed after casing at 2nd break** assessed which at least half of tray is of harvestable maturity for 2nd break, tray-level measurement Days to Pinset visually number of days elapsed after casing at before 1st break assessed which pins first reach 5 mm, tray-level measurement Percent Harvested calculated % of yield harvested by a designated date 17 days after Speed ratio within break, casing (1st yield at date within break/total yield for break), 24 days break, trial-level measurement after casing (2nd break), 32 days after casing (3rd break) Relative Cap Flesh calculated Cap Flesh Thickness Value/Cap 1st break**, Thickness ratio Diameter Value, individual mushroom- 2nd break** level measurement Relative Cap Height calculated Cap Height Value/Cap Diameter Value, 1st break**, ratio individual mushroom-level measurement 2nd break** Relative Stipe Diameter calculated Stipe Diameter Value/Cap Diameter 1st break**, ratio Value, individual mushroom-level 2nd break** measurement Stipe Diameter hand- millimeters (mm), stipe diameter at base 1st break**, measurement of cap, individual mushroom-level 2nd break** measurement Tray Coverage visually 0 to 5 (from no coverage to full 1st break**, assessed coverage), tray-level measurement 2nd break** Yield (Each Break) weighing kg/m.sup.2, tray-level measurement; or Assessed lb/ft.sup.2, tray-level measurement cumulatively throughout 1st, 2nd, and 3rd breaks Yield (Total) weighing kg/m.sup.2, tray-level measurement; or Assessed lb/ft.sup.2, tray-level measurement cumulatively across 1st, 2nd, and 3rd breaks **at date when at least half of tray is of harvestable maturity in break
Objective description of the variety RO10425
[0037] The following detailed description sets forth the breeding procedures and the characteristics of new cultivar designated RO10425. The strain is maintained and propagated as vegetative mycelium. Spawn (inoculum for the substrate compost) is prepared from the vegetative mycelium. The variety has shown uniformity and stability for the traits, within the limits of expected environmental influence for the traits.
[0038] All homokaryons utilized are strains of Agaricus bisporus.
[0039] Two initial homokaryotic strains were used for breeding. Wild brown strain AA-0096 was crossed with a commercial off-white hybrid mushroom strain.
[0040] More specifically, in a first cross, an AA-0096 slow growing SSI (sg) was crossed with a commercial off-white hybrid protoplast (p).
[0041] A progeny homokaryon resultant from this cross was then selected and backcrossed to the commercial off-white hybrid parent strain. A homokaryon produced by the backcrossing was then selected and crossed to a homokaryon of a bridging cross strain. This cross produced RO10425.
[0042] There are multiple characteristics of RO10425 distinguishable from other varieties, such as a different cap color, rounder cap shape, thicker cap apex flesh, thicker cap edge flesh, and higher or as high yield at second break compared to other available brown commercial strains. Other characteristics of RO10425 distinguishable from other varieties are described below (compared to commercial line BR06, commercial line B14528, and related variety RO14176).
[0043]
Comparison to Other Mushroom Varieties
Initial Trials
Tray-Level Comparisons
[0044] In further evidence of the distinctness and superiority of mushroom strain RO10425, provided below are the results of trials conducted across multiple strains' yielded trays of mushrooms. These tray trials compare yield at each break. Characteristics were measured from 15 trays of strain RO10425 mushrooms and 23 trays of strain BR06 mushrooms (Table 2A and 2B), and corresponding measurements were taken from 24 trays of strain RO14176 mushrooms (Table 3A and 3B).
[0045] Table 2A below provides qualitative comparisons between mushroom strain RO10425 and strain BR06. Column 1 lists the tray-level characteristic (as described in Table 1), column 2 shows the qualitative descriptor for the characteristic for mushroom strain RO10425 relative to BR06, and column 3 shows the qualitative descriptor for mushroom strain BR06 relative to RO10425.
TABLE-US-00002 TABLE 2A Characteristic RO10425 BR06 First Break Yield (lb/ft.sup.2) Lower Higher Second Break Yield (lb/ft.sup.2) Similar Similar Third Break Yield (lb/ft.sup.2) Higher Lower All Breaks Combined Yield (lb/ft.sup.2) Similar Similar
[0046] Table 2B below provides quantitative comparisons between mushroom strain RO10425 and strain BR06. Column 1 lists the tray-level characteristics (as described in Table 1), column 2 shows the average value of the characteristic for mushroom strain RO10425, column 3 shows the standard deviation of the characteristic for mushroom strain RO10425, column 4 shows the average value of the characteristic for mushroom strain BR06, column 5 shows the standard deviation of the characteristic for mushroom strain BR06, and column 6 shows the p-values of two-sample, two-sided t-tests. Bolded p-values indicate p<0.05.
TABLE-US-00003 TABLE 2B RO10425 BR06 Characteristic Average St. Dev. Average St. Dev. p-value First Break Yield (lb/ft.sup.2) 1.61 0.64 2.54 1.29 1.33E02 Second Break Yield (lb/ft.sup.2) 1.69 0.48 1.47 0.51 1.92E01 Third Break Yield (lb/ft.sup.2) 1.32 0.41 0.75 0.50 8.12E04 All Breaks Combined Yield (lb/ft.sup.2) 4.62 0.61 4.76 1.24 6.79E01
[0047] Table 3A below provides qualitative comparisons between mushroom strain RO10425 and strain RO14176. Column 1 lists the tray-level characteristic (as described in Table 1), column 2 shows the qualitative descriptor for the characteristic for mushroom strain RO10425 relative to RO14176, and column 3 shows the qualitative descriptor for mushroom strain RO14176 relative to RO10425.
TABLE-US-00004 TABLE 3A Characteristic RO10425 RO14176 First Break Yield (lb/ft.sup.2) Lower Higher Second Break Yield (lb/ft.sup.2) Similar Similar Third Break Yield (lb/ft.sup.2) Higher Lower All Breaks Combined Yield (lb/ft.sup.2) Lower Higher
[0048] Table 3B below provides quantitative comparisons between mushroom strain RO10425 and strain BR06. Column 1 lists the tray-level characteristics (as described in Table 1), column 2 shows the average value of the characteristic for mushroom strain RO10425, column 3 shows the standard deviation of the characteristic for mushroom strain RO10425, column 4 shows the average value of the characteristic for mushroom strain RO14176, column 5 shows the standard deviation of the characteristic for mushroom strain RO14176, and column 6 shows the p-values of two-sample, two-sided t-tests. Bolded p-values indicate p <0.05.
TABLE-US-00005 TABLE 3B RO10425 RO14176 Characteristic Average St. Dev. Average St. Dev. p-value First Break Yield (lb/ft.sup.2) 1.61 0.64 2.54 0.83 7.01E04 Second Break Yield (lb/ft.sup.2) 1.69 0.48 1.91 0.52 1.91E01 Third Break Yield (lb/ft.sup.2) 1.32 0.41 0.92 0.38 3.41E03 All Breaks Combined Yield (lb/ft.sup.2) 4.62 0.61 5.37 1.08 1.94E02
Individual Mushroom-Level Comparisons
[0049] In additional evidence of the distinctness and superiority of mushroom strain RO10425, provided below is a summary of precise quantitative measurements of individual mushrooms at first and second breaks. These data compare Cap L* Value, Cap a* Value, and Cap b* Values in the International Commission on Illumination (CIELAB) color space, Cap Diameter, Cap Height, Cap Flesh Thickness, Stipe Diameter, Relative Cap Height, Relative Cap Flesh Thickness, and Relative Stipe Diameter (as described in Table 1). Measurements from 49 first break and 37 second break RO10425 mushrooms were compared to 80 first break and 40 second break BR06 mushrooms (Tables 4A and 4B).
[0050] Table 4A below provides qualitative comparisons between mushroom strain RO10425 and strain BR06. Column 1 lists the mushroom-level characteristics (as described in Table 1), column 2 shows the qualitative descriptor for the characteristic for mushroom strain RO10425 relative to BR06, and column 3 shows the qualitative descriptor for mushroom strain BR06 relative to RO10425.
TABLE-US-00006 TABLE 4A Characteristic RO10425 BR06 First Break Cap L* Value (dark-light) Lighter Darker Cap a* Value (green-red) Greener Redder Cap b* Value (blue-yellow) Bluer Yellower Cap Diameter Similar Similar Cap Height Similar Similar Cap Flesh Thickness Similar Similar Stipe Diameter Larger Smaller Relative Cap Height Similar Similar Relative Cap Flesh Thickness Thicker Thinner Relative Stipe Diameter Larger Smaller Second Break Cap L* Value (dark-light) Lighter Darker Cap a* Value (green-red) Greener Redder Cap b* Value (blue-yellow) Bluer Yellower Cap Diameter Larger Smaller Cap Height Taller Shorter Cap Flesh Thickness Thicker Thinner Stipe Diameter Larger Smaller Relative Cap Height Similar Similar Relative Cap Flesh Thickness Similar Similar Relative Stipe Diameter Similar Similar
[0051] Table 4B below provides quantitative comparisons between mushroom strain RO10425 and strain BR06. Column 1 lists the mushroom-level characteristics (as described in Table 1), column 2 shows the average value of the characteristic for mushroom strain RO10425, column 3 shows the standard deviation of the characteristic for mushroom strain RO10425, column 4 shows the average value of the characteristic for mushroom strain BR06, column 5 shows the standard deviation of the characteristic for mushroom strain BR06, and column 6 shows the p-values of two-sample, two-sided t-tests. Bolded p-values indicate p <0.05.
TABLE-US-00007 TABLE 4B RO10425 BR06 St. St. Characteristic Average Dev. Average Dev. p-value First Break Cap L* Value (dark-light) 65.84 5.15 62.20 5.58 3.22E04 Cap a* Value (green-red) 7.51 0.99 9.43 1.15 5.94E17 Cap b* Value (blue-yellow) 26.03 1.29 28.67 1.63 7.91E17 Cap Diameter 43.45 3.41 44.05 3.80 3.70E01 Cap Height 27.09 1.99 27.45 1.99 3.27E01 Cap Flesh Thickness 15.03 1.42 14.57 1.46 8.09E02 Stipe Diameter 21.66 2.08 20.03 2.08 3.18E05 Relative Cap Height 0.62 0.03 0.62 0.04 9.82E01 Relative Cap Flesh Thickness 0.35 0.04 0.33 0.03 1.61E02 Relative Stipe Diameter 0.56 0.05 0.53 0.03 1.02E03 Second Break Cap L* Value (dark-light) 64.44 2.79 62.19 3.79 4.12E03 Cap a* Value (green-red) 7.05 0.79 7.99 1.07 3.86E05 Cap b* Value (blue-yellow) 26.48 1.15 27.01 1.06 3.64E02 Cap Diameter 45.20 3.51 41.98 4.43 7.39E04 Cap Height 28.78 2.27 26.81 2.89 1.42E03 Cap Flesh Thickness 15.19 1.61 14.30 1.53 1.54E02 Stipe Diameter 20.90 2.02 18.74 3.24 8.38E04 Relative Cap Height 0.64 0.04 0.64 0.03 8.37E01 Relative Cap Flesh Thickness 0.34 0.04 0.34 0.02 5.52E01 Relative Stipe Diameter 0.46 0.04 0.45 0.05 8.97E02
[0052] Additional analyses were performed on the measurements taken across individual mushrooms of each variety at multiple breaks, in order to compare more generalized mushroom traits irrespective of whether the mushrooms belonged to the first vs. second break of harvesting. The same characteristics described above (see Table 1) were measured from 86 mushrooms of strain RO10425 across the first and second break, and corresponding measurements were taken from 150 mushrooms of strain BR06 across the first and second break (Table 5A and 5B).
[0053] Table 5A below provides qualitative comparisons between mushroom strain RO10425 and strain BR06 made irrespective of break. Column 1 lists the mushroom-level characteristics (as described in Table 1), column 2 shows the qualitative descriptor for the characteristic for mushroom strain RO10425 relative to BR06, and column 3 shows the qualitative descriptor for mushroom strain BR06 relative to RO10425.
TABLE-US-00008 TABLE 5A Characteristic RO10425 BR06 Cap L* Value (dark-light) Lighter Darker Cap a* Value (green-red) Greener Redder Cap b* Value (blue-yellow) Bluer Yellower Cap Diameter Larger Smaller Cap Height Taller Shorter Cap Flesh Thickness Thicker Thinner Stipe Diameter Larger Smaller Relative Cap Height Taller Shorter Relative Cap Flesh Thickness Thicker Thinner Relative Stipe Diameter Larger Smaller
[0054] Table 5B below provides quantitative comparisons between mushroom strain RO10425 and strain BR06. Column 1 lists the mushroom-level characteristics (as described in Table 1), column 2 shows the average value of the characteristic for mushroom strain RO10425, column 3 shows the standard deviation of the characteristic for mushroom strain RO10425, column 4 shows the average value of the characteristic for mushroom strain BR06, column 5 shows the standard deviation of the characteristic for mushroom strain BR06, and column 6 shows the p-values of two-sample, two-sided t-tests. Bolded p-values indicate p <0.05.
TABLE-US-00009 TABLE 5B RO10425 BR06 Characteristic Average St. Dev. Average St. Dev. p-value Cap L* Value (dark-light) 65.24 4.33 62.86 4.95 2.49E04 Cap a* Value (green-red) 7.31 0.93 8.77 1.31 3.90E17 Cap b* Value (blue-yellow) 26.22 1.24 28.31 1.61 5.99E21 Cap Diameter 44.20 3.55 42.84 4.00 9.33E03 Cap Height 27.82 2.26 26.53 2.59 1.55E04 Cap Flesh Thickness 15.10 1.50 14.01 1.68 1.12E06 Stipe Diameter 21.33 2.07 18.70 2.98 6.06E12 Relative Cap Height 0.63 0.03 0.62 0.04 4.84E02 Relative Cap Flesh Thickness 0.34 0.04 0.33 0.03 9.34E04 Relative Stipe Diameter 0.48 0.04 0.44 0.05 1.96E12
[0055] Tables 6A and 6B compare instant strain RO10425 and related strain RO14176. Table 6A below provides qualitative comparisons between mushroom strain RO10425 and strain RO14176. Measurements from 49 first break and 37 second break RO10425 mushrooms were compared to 80 first break and 40 second break RO14176 mushrooms (Tables 6A and 6B). Column 1 lists the mushroom-level characteristics (as described in Table 1), column 2 shows the qualitative descriptor for the characteristic for mushroom strain RO10425 relative to RO14176, and column 3 shows the qualitative descriptor for mushroom strain RO14176 relative to RO10425.
TABLE-US-00010 TABLE 6A Characteristic RO10425 RO14176 First Break Cap L* Value (dark-light) Similar Similar Cap a* Value (green-red) Similar Similar Cap b* Value (blue-yellow) Similar Similar Cap Diameter Similar Similar Cap Height Shorter Taller Stipe Diameter Larger Smaller Relative Cap Height Shorter Taller Relative Cap Flesh Similar Similar Thickness Relative Stipe Diameter Larger Smaller Second Break Cap L* Value (dark-light) Similar Similar Cap a* Value (green-red) Redder Greener Cap b* Value (blue-yellow) Yellower Bluer Cap Diameter Larger Smaller Cap Height Similar Similar Stipe Diameter Larger Smaller Relative Cap Height Shorter Taller Relative Cap Flesh Thinner Thicker Thickness Relative Stipe Diameter Similar Similar
[0056] Table 6B below provides quantitative comparisons between mushroom strain RO10425 and strain RO14176. Column 1 lists the mushroom-level characteristics (as described in Table 1), column 2 shows the average value of the characteristic for mushroom strain RO10425, column 3 shows the standard deviation of the characteristic for mushroom strain RO10425, column 4 shows the average value of the characteristic for mushroom strain RO14176, column 5 shows the standard deviation of the characteristic for mushroom strain RO14176, and column 6 shows the p-values of two-sample, two-sided t-tests. Bolded p-values indicate p <0.05.
TABLE-US-00011 TABLE 6B RO10425 RO14176 Characteristic Average St. Dev. Average St. Dev. p-value First Break Cap L* Value (dark-light) 65.84 5.15 65.91 4.79 9.38E01 Cap a* Value (green-red) 7.51 0.99 7.49 1.05 9.32E01 Cap b* Value (blue-yellow) 26.03 1.29 26.35 1.35 1.82E01 Cap Diameter 43.45 3.41 43.53 3.27 9.01E01 Cap Height 27.09 1.99 27.89 2.12 3.55E02 Cap Flesh Thickness 15.03 1.42 15.33 1.36 2.42E01 Stipe Diameter 21.66 2.08 19.99 1.90 7.06E06 Relative Cap Height 0.62 0.03 0.64 0.03 2.43E03 Relative Cap Flesh Thickness 0.35 0.04 0.35 0.03 3.25E01 Relative Stipe Diameter 0.50 0.04 0.46 0.04 4.71E07 Second Break Cap L* Value (dark-light) 64.44 2.79 65.61 3.67 1.24E01 Cap a* Value (green-red) 7.05 0.79 6.36 0.76 2.32E04 Cap b* Value (blue-yellow) 26.48 1.15 25.05 1.46 1.07E05 Cap Diameter 45.20 3.51 41.88 2.68 1.23E05 Cap Height 28.78 2.27 28.22 2.12 2.65E01 Cap Flesh Thickness 15.19 1.61 15.75 1.61 1.31E01 Stipe Diameter 20.90 2.02 19.16 2.29 6.92E04 Relative Cap Height 0.64 0.04 0.67 0.04 7.23E05 Relative Cap Flesh Thickness 0.34 0.04 0.38 0.04 1.79E05 Relative Stipe Diameter 0.46 0.04 0.46 0.05 6.21E01
[0057]
[0058] Additional analysis was conducted for measurements taken across individual mushrooms of each variety at multiple breaks, in order to compare more generalized mushroom traits irrespective of whether the mushrooms belonged to the first vs. second break of harvesting. The same characteristics described above (see Table 1) were measured from 86 mushrooms of strain RO10425 across the first and second break, and corresponding measurements were taken from 170 mushrooms of strain RO14176 across the first and second break (Table 7A and 7B). Table 7A below provides qualitative comparisons between mushroom strain RO10425 and strain RO14176. Column 1 lists the mushroom-level characteristics (as described in Table 1), column 2 shows the qualitative descriptor for the characteristic for mushroom strain RO10425 relative to RO14176, and column 3 shows the qualitative descriptor for mushroom strain RO14176 relative to RO10425.
TABLE-US-00012 TABLE 7A Characteristic RO10425 RO14176 Cap L* Value (dark-light) Darker Lighter Cap a* Value (green-red) Redder Greener Cap b* Value (blue-yellow) Similar Similar Cap Diameter Larger Shorter Cap Height Taller Shorter Cap Flesh Thickness Similar Similar Stipe Diameter Larger Smaller Relative Cap Height Shorter Taller Relative Cap Flesh Thickness Similar Similar Relative Stipe Diameter Larger Smaller
[0059] Table 7B below provides quantitative comparisons between mushroom strain RO10425 and strain RO14176. Column 1 lists the mushroom-level characteristics (as described in Table 1), column 2 shows the average value of the characteristic for mushroom strain RO10425, column 3 shows the standard deviation of the characteristic for mushroom strain RO10425, column 4 shows the average value of the characteristic for mushroom strain RO14176, column 5 shows the standard deviation of the characteristic for mushroom strain RO14176, and column 6 shows the p-values of two-sample, two-sided t-tests. Bolded p-values indicate p <0.05.
TABLE-US-00013 TABLE 7B RO10425 RO14176 Characteristic Average St. Dev. Average St. Dev. p-value Cap L* Value (dark-light) 65.24 4.33 67.15 4.61 1.60E03 Cap a* Value (green-red) 7.31 0.93 6.93 1.09 6.45E03 Cap b* Value (blue-yellow) 26.22 1.24 26.44 1.59 2.60E01 Cap Diameter 44.20 3.55 42.36 3.30 5.22E05 Cap Height 27.82 2.26 27.14 2.37 2.88E02 Cap Flesh Thickness 15.10 1.50 14.70 1.84 7.93E02 Stipe Diameter 21.33 2.07 18.53 2.68 1.68E15 Relative Cap Height 0.63 0.03 0.64 0.04 2.54E02 Relative Cap Flesh Thickness 0.34 0.04 0.35 0.04 3.75E01 Relative Stipe Diameter 0.48 0.04 0.44 0.05 2.98E11
Additional Trials
Tray-Level Comparisons
[0060] Several additional trials were conducted for characterizing RO10425. Table 8 below provides qualitative comparisons between mushroom strain RO10425 and strain BR06. Traits of particular interest are bolded. Column 1 lists the tray-level characteristics (as described in Table 1), column 2 shows the qualitative descriptor for the characteristic for mushroom strain RO10425 relative to BR06, and column 3 shows the qualitative descriptor for mushroom strain BR06 relative to RO10425.
TABLE-US-00014 TABLE 8 Characteristic RO10425 BR06 Before First Break Days to Pinset Similar Similar First Break Days to 1st Break Similar Similar Percent Harvested Speed Similar Similar Yield (kg/m)2 Similar Similar Second Break Days to 2nd Break Similar Similar Yield (kg/m)2 Higher Lower Third Break Yield (kg/m)2 Similar Similar All Breaks Combined Yield (kg/m)2 Similar Similar
[0061] The calculations that informed these qualitative descriptors are shown in Table 10.
[0062] Table 9 below provides qualitative comparisons between mushroom strain RO10425 and strain B14528. Column 1 lists the tray-level characteristic (as described in Table 1), column 2 shows the qualitative descriptor for the characteristic for mushroom strain RO10425 relative to B14528, and column 3 shows the qualitative descriptor for mushroom strain B14528 relative to RO10425.
TABLE-US-00015 TABLE 9 Characteristic RO10425 B14528 Before First Break Days to Pinset Similar Similar First Break Days to 1st Break Similar Similar Percent Harvested Speed Similar Similar Yield (kg/m)2 Similar Similar Second Break Days to 2nd Break Similar Similar Yield (kg/m)2 Similar Similar Third Break Yield (kg/m)2 Similar Similar All Breaks Combined Yield (kg/m)2 Similar Similar
[0063] The calculations that informed these qualitative descriptors are shown in Table 10.
[0064] The first part of analyzing trait measurements in the additional trials herein included making comparisons between all three of instant strain RO10425, comparative strain BR06, and comparative strain B14528. For these three strains, a Shapiro-Wilk test and a Levene test were performed first to assess if the data satisfy the assumptions of normality and equal variances among groups, respectively, of a standard ANOVA. The Shapiro-Wilk test and Levene test determined that most of these traits satisfied neither assumption. Therefore, a Kruskal-Wallis test (Kruskal, W. H., & Wallis, W. A. (1952). Use of ranks in one-criterion variance analysis. Journal of the American statistical Association, 47(260), 583-621), a non-parametric variation of ANOVA that does not assume normality, equal variances among groups, or equal sample sizes among groups, was performed. Medians and interquartile ranges (IQR) were reported, as these metrics are robust to non-normal distributions. Means are also reported for consistency with normally-distributed traits.
[0065] The Shapiro-Wilk test and Levene test determined that most developmental timing traits (Days to Pinset, Days to 1st Break, and Days to 2nd Break) and yield traits were not normally distributed and variances among groups were not equal. The Kruskal-Wallis test and Dunn post-hoc test (Dunn's Test; Dunn, O. J. (1964). Multiple comparisons using rank sums. Technometrics, 6(3), 241-252) were used in these cases. When the Kruskal-Wallis test results were significant based on a threshold of p<0.05, Dunn's test was performed post-hoc to make pairwise strain comparisons (Dunn, 1964). P-values were adjusted using the Bonferroni correction to minimize risk of Type 1 error (false significance). An exception to this trend was Percent Harvested Speed, which was normally distributed and exhibited equal variance for all strains. For assessing Percent Harvested Speed, a standard ANOVA was used and a Tukey Honestly Significant Difference (HSD) test was performed post-hoc for pairwise strain comparisons. Standard deviations multiplied by 100 for consistency alongside percentages (SD*100) were reported for this trait. This testing was performed on groups of three strains, as shown in Table 10, but the other results herein using this test show a pairwise subset of the tests for consistency with the initial trials.
[0066] In Table 10, column 1 shows each trait and the strains compared for said trait, column 2 shows the mean value for that strain's measurements of that trait, column 3 shows the median value of the same measurements, column 4 shows the interquartile range (IQR) for the same measurements, column 5 shows the number of trials conducted, and column 6 shows the statistical significance group calculated for that strain for that trait according to Dunn's Test. Traits of particular interest are bolded.
TABLE-US-00016 TABLE 10 Number Significance of Group Variety Mean Median IQR Trials (Dunn's Test) Before First Break Days to Pinset RO10425 12.00 12.00 2.00 4 a B14528 12.00 12.00 1.75 4 a BR06 12.40 12.00 1.00 4 a First Break Days to 1st Break RO10425 18.00 18.00 1.00 4 a B14528 17.60 18.00 1.75 4 a BR06 18.10 18.00 1.00 4 a Yield (kg/m)2 RO10425 10.64 10.64 1.76 3 a B14528 9.81 9.52 4.44 3 a BR06 10.30 8.35 5.57 3 a Second Break Days to 2nd Break RO10425 24.90 25.00 1.75 4 a B14528 24.80 25.00 1.00 4 a BR06 25.30 26.00 1.00 4 a Yield (kg/m)2 RO10425 8.74 8.10 3.27 3 a B14528 7.13 6.93 1.51 3 ab BR06 6.20 5.86 1.61 3 a Third Break Yield (kg/m)2 RO10425 3.66 3.52 0.78 3 ab B14528 5.81 5.42 1.81 3 b BR06 2.64 1.95 2.10 3 a All Breaks Combined Yield (kg/m)2 RO10425 23.05 23.14 4.25 3 a B14528 22.75 22.61 2.29 3 a BR06 19.14 18.70 5.52 3 a Significance Percent Harvested Speed Group (measured in First Break) SD * 100 (Tukey HSD) RO10425 36.7% 34.8% 32.70 3 a B14528 36.2% 45.2% 25.00 3 a BR06 28.9% 12.9% 37.70 3 a
Individual Mushroom-Level Comparisons
[0067] In additional evidence of the distinctness and superiority of mushroom strain RO10425, provided below is a summary of precise quantitative measurements of individual mushrooms at first and second breaks. These data compare Cap L* Value, Cap a* Value, and Cap b* Values in the International Commission on Illumination (CIELAB) color space, Cap Apex Flesh Thickness, Cap Edge Flesh Thickness, and Cap Roundness (Calculated) (as described in Table 1). Measurements were taken for hundreds of mushrooms per trait, noted as sample size (n) in Tables 11B and 12B.
[0068] Table 11A below provides qualitative comparisons between mushroom strain RO10425 and strain BR06. Traits of particular interest are bolded. Column 1 lists the mushroom-level characteristics (as described in Table 1), column 2 shows the qualitative descriptor for the characteristic for mushroom strain RO10425 relative to BR06, and column 3 shows the qualitative descriptor for mushroom strain BR06 relative to RO10425.
TABLE-US-00017 TABLE 11A Characteristic RO10425 BR06 First Break Cap L* Value (dark-light) Lighter Darker Cap a* Value (green-red) Greener Redder Cap b* Value (blue-yellow) Bluer Yellower Cap Apex Flesh Thickness Thicker Thinner Cap Edge Flesh Thickness Thicker Thinner Cap Roundness (Calculated) Rounder Flatter Second Break Cap L* Value (dark-light) Lighter Darker Cap a* Value (green-red) Greener Redder Cap b* Value (blue-yellow) Bluer Yellower Cap Apex Flesh Thickness Thicker Thinner Cap Edge Flesh Thickness Thicker Thinner Cap Roundness (Calculated) Rounder Flatter
[0069] Table 11B shows additional quantitative comparisons between mushroom strain RO10425 and strain BR06. Column 1 shows each trait and the strains compared for said trait, column 2 shows the mean value for that strain's measurements of that trait, column 3 shows the median value of the same measurements, column 4 shows the interquartile range (IQR) for the same measurements, column 5 shows the number of trials conducted, and column 6 shows the statistical significance group calculated for that strain for that trait according to Dunn's Test.
TABLE-US-00018 TABLE 11B Number of Significance Mushrooms Group Variety Mean Median IQR (n) (Dunn's Test) First Break Cap L* Value (dark-light) RO10425 65.66 65.22 7.34 209 a BR06 63.30 62.74 9.83 797 b Cap a* Value (green-red) RO10425 7.35 7.43 1.97 209 a BR06 8.67 8.96 2.58 787 b Cap b* Value (blue-yellow) RO10425 28.43 28.58 3.10 209 a BR06 28.72 29.04 3.17 776 b Cap Apex Flesh Thickness RO10425 0.55 0.55 0.04 166 a BR06 0.52 0.52 0.05 541 b Cap Edge Flesh Thickness RO10425 0.30 0.31 0.05 166 a BR06 0.21 0.21 0.09 537 b Cap Roundness (Calculated) RO10425 0.64 0.63 0.05 166 a BR06 0.61 0.61 0.04 541 b Second Break Cap L* Value (dark-light) RO10425 68.82 69.47 8.20 157 a BR06 65.68 65.49 5.90 340 b Cap a* Value (green-red) RO10425 6.15 6.26 2.55 157 a BR06 7.21 7.31 2.42 340 b Cap b* Value (blue-yellow) RO10425 27.69 27.46 2.54 157 a BR06 28.15 28.49 2.86 340 b Cap Apex Flesh Thickness RO10425 0.55 0.55 0.04 157 a BR06 0.51 0.52 0.07 362 b Cap Edge Flesh Thickness RO10425 0.32 0.32 0.05 157 a BR06 0.23 0.23 0.09 362 b Cap Roundness (Calculated) RO10425 0.61 0.61 0.05 157 a BR06 0.60 0.59 0.06 363 b
[0070] Table 12A below provides qualitative comparisons between mushroom strain RO10425 and strain B14528. Traits of interest are bolded in Tables 12A and 12B. Column 1 lists the individual mushroom-level characteristic (as described in Table 1), column 2 shows the qualitative descriptor for the characteristic for mushroom strain RO10425 relative to B14528, and column 3 shows the qualitative descriptor for mushroom strain B14528 relative to RO10425.
TABLE-US-00019 TABLE 12A Characteristic RO10425 B14528 First Break Cap L* Value (dark-light) Lighter Darker Cap a* Value (green-red) Greener Redder Cap b* Value (blue-yellow) Bluer Yellower Cap Apex Flesh Thickness Thicker Thinner Cap Edge Flesh Thickness Thicker Thinner Cap Roundness (Calculated) Rounder Flatter Second Break Cap L* Value (dark-light) Similar Similar Cap a* Value (green-red) Similar Similar Cap b* Value (blue-yellow) Similar Similar Cap Apex Flesh Thickness Thicker Thinner Cap Edge Flesh Thickness Thicker Thinner Cap Roundness (Calculated) Rounder Flatter
[0071] Table 12B below provides quantitative comparisons between mushroom strain RO10425 and strain B14528, which resulted in the qualitative descriptors of Table 12A. Column 1 shows each individual mushroom-level trait (as described in Table 1) and the strains compared for said trait, column 2 shows the mean value for that strain's measurements of that trait, column 3 shows the median value of the same measurements, column 4 shows the interquartile range (IQR) for the same measurements, column 5 shows the number of mushrooms assessed (n), and column 6 shows the statistical significance group calculated for that strain for that trait according to Dunn's Test.
TABLE-US-00020 TABLE 12B Number of Significance Mushrooms Group Variety Mean Median IQR (n) (Dunn's Test) First Break Cap L* Value (dark-light) RO10425 65.66 65.22 7.34 209 a B14528 64.06 64.13 6.56 340 c Cap a* Value (green-red) RO10425 7.35 7.43 1.97 209 a B14528 8.26 8.31 2.13 340 c Cap b* Value (blue-yellow) RO10425 28.43 28.58 3.10 209 a B14528 30.31 30.16 3.39 340 c Cap Apex Flesh Thickness RO10425 0.55 0.55 0.04 166 a B14528 0.47 0.46 0.06 387 c Cap Edge Flesh Thickness RO10425 0.30 0.31 0.05 166 a B14528 0.22 0.22 0.08 387 b Cap Roundness (Calculated) RO10425 0.64 0.63 0.05 166 a B14528 0.61 0.61 0.05 389 b Second Break Cap L* Value (dark-light) RO10425 68.82 69.47 8.20 157 a B14528 68.56 69.02 6.54 170 a Cap a* Value (green-red) RO10425 6.15 6.26 2.55 157 a B14528 6.09 5.96 2.05 170 a Cap b* Value (blue-yellow) RO10425 27.69 27.46 2.54 157 a B14528 27.37 27.78 3.22 170 a Cap Apex Flesh Thickness RO10425 0.55 0.55 0.04 157 a B14528 0.46 0.46 0.05 198 c Cap Edge Flesh Thickness RO10425 0.32 0.32 0.05 157 a B14528 0.23 0.23 0.06 198 b Cap Roundness (Calculated) RO10425 0.61 0.61 0.05 157 a B14528 0.60 0.60 0.05 198 b
[0072] As conducted in initial trials, comparisons were made between instant strain RO10425 and related strain RO14176. Tables 13A below provides qualitative comparisons between mushroom strain RO10425 and strain RO14176. Column 1 lists the mushroom-level characteristics (as described in Table 1), column 2 shows the qualitative descriptor for the characteristic for mushroom strain RO10425 relative to RO14176, and column 3 shows the qualitative descriptor for mushroom strain RO14176 relative to RO10425.
TABLE-US-00021 TABLE 13A Characteristic RO10425 RO14176 First Break Cap L* Value (dark-light) Darker Lighter Cap a* Value (green-red) Redder Greener Cap b* Value (blue-yellow) Similar Similar Cap Diameter Similar Similar Cap Height Similar Similar Cap Flesh Thickness Similar Similar Stipe Diameter Larger Smaller Relative Cap Height Similar Similar Relative Cap Flesh Thickness Similar Similar Relative Stipe Diameter Larger Smaller Second Break Cap L* Value (dark-light) Similar Similar Cap a* Value (green-red) Similar Similar Cap b* Value (blue-yellow) Similar Similar Cap Diameter Similar Similar Cap Height Similar Similar Cap Flesh Thickness Similar Similar Stipe Diameter Larger Smaller Relative Cap Height Similar Similar Relative Cap Flesh Thickness Similar Similar Relative Stipe Diameter Larger Smaller
[0073] Table 13B below provides quantitative comparisons between mushroom strain RO10425 and strain RO14176 that informed the qualitative comparisons of Table 13A. In Table 13B, Column 1 lists the mushroom-level characteristics (as described in Table 1), column 2 shows the average value of the characteristic for mushroom strain RO10425, column 3 shows the standard deviation of the characteristic for mushroom strain RO10425, column 4 shows the average value of the characteristic for mushroom strain RO14176, column 5 shows the standard deviation of the characteristic for mushroom strain RO14176, and column 6 shows the p-values of two-sample, two-sided t-tests. Bolded p-values indicate p<0.05.
TABLE-US-00022 TABLE 13B RO10425 RO14176 Characteristic Average St. Dev. Average St. Dev. p-value First Break Cap L* Value (dark-light) 65.17 5.27 66.97 4.84 1.18E05 Cap a* Value (green-red) 7.55 1.68 6.89 1.63 6.95E07 Cap b* Value (blue-yellow) 28.52 2.09 28.46 2.07 6.92E01 Cap Diameter 43.18 3.81 43.25 3.94 8.21E01 Cap Height 27.67 2.65 27.87 2.19 3.44E01 Cap Flesh Thickness 15.26 1.61 15.19 1.73 6.44E01 Stipe Diameter 21.66 2.08 19.92 2.43 1.94E06 Relative Cap Height 0.65 0.04 0.65 0.04 3.41E01 Relative Cap Flesh Thickness 0.35 0.03 0.35 0.05 1.66E01 Relative Stipe Diameter 0.50 0.04 0.45 0.05 1.50E09 Second Break Cap L* Value (dark-light) 68.82 5.25 69.32 4.59 3.11E01 Cap a* Value (green-red) 6.15 1.63 6.02 1.64 4.26E01 Cap b* Value (blue-yellow) 27.69 1.91 27.50 2.11 3.19E01 Cap Diameter 43.19 4.08 42.56 3.62 1.08E01 Cap Height 26.65 2.49 26.28 2.29 1.31E01 Cap Flesh Thickness 14.51 1.34 14.24 1.60 6.04E02 Stipe Diameter 20.90 2.02 17.80 2.58 4.81E11 Relative Cap Height 0.62 0.04 0.62 0.05 3.57E01 Relative Cap Flesh Thickness 0.34 0.03 0.34 0.04 9.52E01 Relative Stipe Diameter 0.46 0.04 0.42 0.05 1.84E06
[0074] One of the advantages of the strain RO10425 is its incompatibility with other commercial strains. Table 14 below provides results of compatibility tests between strain RO10425 and commercial strain BR06, commercial strain B14528, related strain RO12756, and related strain RO14176. These results indicate that strain RO10425 is incompatible with strain BR06, strain B14528, and related strain RO12756, while strain RO10425 is compatible with related strain RO14176. This was true regardless of which tested strain served as the compost variety and which tested strain served as the casing variety.
TABLE-US-00023 TABLE 14 Strains (compost variety/ Mean Median SD Significance casing variety) lb/ft.sup.2 kg/m.sup.2 lb/ft.sup.2 kg/m.sup.2 lb/ft.sup.2 kg/m.sup.2 Trials Group RO10425/RO10425 2.71 13.23 2.86 13.94 0.50 2.44 3 a RO12756/RO12756 2.62 12.79 2.61 12.74 0.41 2.00 3 a RO14176/RO14176 2.66 12.99 2.41 11.77 0.61 2.98 3 a BR06/BR06 2.62 12.79 2.46 11.99 1.15 5.61 2 a B14528/B14528 2.93 14.31 2.92 14.23 0.34 1.66 1 a RO10425/RO14176, 2.43 11.86 2.75 13.40 0.77 3.76 1 a RO14176/RO10425 RO10425/B14528, 0.71 3.47 0.65 3.15 0.17 0.83 1 b B14528/RO10425 RO12756/RO10425, 0.91 4.44 1.04 5.08 0.35 1.71 1 b RO10425/RO12756 RO10425/BR06, 0.80 3.91 0.92 4.47 0.51 2.49 1 b BR06/RO10425