USPP35511P3 - Bunashimeji mushroom named ‘HKHM25’ - Google Patents

Bunashimeji mushroom named ‘HKHM25’ Download PDF

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USPP35511P3
USPP35511P3 US17/803,159 US202217803159V USPP35511P3 US PP35511 P3 USPP35511 P3 US PP35511P3 US 202217803159 V US202217803159 V US 202217803159V US PP35511 P3 USPP35511 P3 US PP35511P3
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hkhm25
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bunashimeji
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Ayumi Nakagoshi
Aya Seki
Chiaki Goto
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Hokuto Corp
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H15/00—Fungi; Lichens

Definitions

  • This invention relates to a new and distinct variety of Bunashimeji mushroom, Hypsizygus marmoreus (Peck) H.E.Bigelow.
  • This new variety named ‘HKHM25’, cultivated by repeated breeding of Bunashimeji mushroom having thick and elastic stems, dark cap color, strong cap roll, and a large mushroom size, has a high quality and enhanced cultivation stability, and ensures presentable stability, reproducibility, and uniformity.
  • Bunashimeji mushrooms The annual production of Bunashimeji mushrooms is approximately 120,000 tons in 2020, making it the second most consumed edible mushroom after enokitake mushroom in Japan.
  • Bunashimeji mushrooms have become available in large quantities in the market, Bunashimeji mushrooms with improved flavor, quality and shelf life has been pursued, which led to the development of ‘Hokuto 18gokin’ by our company.
  • our company has been conducting a wide variety of studies for the more stable cultivation and better quality of Bunashimeji mushrooms.
  • Our company has developed a ‘marmo22go’ mushroom which is thicker, whiter, and has larger mushroom than ‘Hokuto 18gokin’.
  • ‘HKHM25’ which has thicker stems and stronger cap roll, as compared to ‘marmo22go’, was developed, confirming its stability, reproducibility, and uniformity.
  • the present invention is a new and distinct variety of mushroom plant characterized particularly by having thick and elastic stems, dark cap color, strong cap roll, and a large mushroom size, which can be cultivated by gathering and repeated breeding of fungal strains having a stability, reproducibility and uniformity when being produced.
  • This novel and distinct variety of mushroom is identified as ‘HKHM25’.
  • FIG. 1 shows a phylogenetic tree illustrating the antecedents of ‘HKHM25’.
  • FIGS. 2 A and 2 B respectively show front and back images of a dual-culture of ‘HKHM25’.
  • FIGS. 3 A and 3 B respectively show front and back images of a dual-culture of ‘HKHM25’ and ‘Hokuto 18gokin’.
  • FIGS. 4 A and 4 B respectively show front and back images of a dual-culture of ‘HKHM25’ and ‘marmo22go’.
  • FIGS. 5 A and 5 B respectively show front and back images of a fungal flora of ‘HKHM25’ grown in a petri dish.
  • FIGS. 6 A and 6 B respectively show front and back image of a fungal flora of ‘Hokuto 18gokin’ grown in a petri dish.
  • FIGS. 7 A and 7 B respectively show front and back images of fungal flora of ‘marmo22go’ grown in a petri dish.
  • FIG. 8 shows an image of cultivation area of ‘HKHM25’.
  • FIG. 9 shows an image of cultivation area of ‘Hokuto 18gokin’.
  • FIG. 10 shows an image of cultivation area of ‘marmo22go’.
  • FIG. 11 shows an image of a fruit body of ‘HKHM25’.
  • FIG. 12 shows an image of a fruit body of ‘Hokuto 18gokin’.
  • FIG. 13 shows an image of a fruit body of ‘marmo22go’.
  • FIG. 14 shows an image of density and surface color of fungal flora in cultured hyphae of ‘HKHM25’.
  • FIG. 15 shows an image of density and surface color of fungal flora in cultured hyphae of ‘Hokuto 18gokin’.
  • FIG. 16 shows an image of density and surface color of fungal flora in cultured hyphae of ‘marmo22go’.
  • FIG. 17 shows an image of development of aerial hypha of ‘HKHM25’.
  • FIG. 18 shows an image of development of aerial hypha of ‘Hokuto 18gokin’.
  • FIG. 19 shows an image of development of aerial hypha of ‘marmo22go’.
  • FIG. 20 shows an image of size and distribution of mottle of ‘HKHM25’.
  • FIG. 21 shows an image of size and distribution of mottle of ‘Hokuto 18gokin’.
  • FIG. 22 shows an image of size and distribution of mottle of ‘marmo22go’.
  • FIG. 23 shows an image of shape and thickness of stipe of ‘HKHM25’.
  • FIG. 24 shows an image of shape and thickness of stipe of ‘Hokuto 18gokin’.
  • FIG. 25 shows an image of shape and thickness of stipe of ‘marmo22go’.
  • ‘HKHM25’ has been asexually reproduced by tissue culture, at HOKUTO CORPORATION in Japan.
  • the history of the ‘HKHM25’ mushroom in terms of improvement period and the like are set forth in the following chronological list of each stage of variety improvement:
  • the ‘HKHM25’ mushroom has the following characteristics: thick and elastic stems, dark cap color, strong cap roll, and overall large size.
  • Dual culture was performed for the ‘HKHM25’ mushroom and a similar variety to examine whether or not a zone line is formed.
  • a potato dextrose agar medium was used, and the ‘HKHM25’ mushroom and the similar variety were inoculated thereon face to face at an interval of 3 cm, and then culture was performed at 25° C. for 28 days to examine whether or not a zone line was formed.
  • preculture was performed at 25° C. for 4 days so as to make the regeneration of hyphae equal (about 10 mm in diameter), and then culture was performed for 7 days at intervals of 5° C. between 5° C. and 30° C.
  • An average daily hyphae growth rate was calculated based on a hyphae growth rate for 7 days of the culture.
  • the optimum culture period for ‘HKHM25’ is 80 days, and the number of growing days at that time is 23.1 days.
  • ‘HKHM25’ had the fastest average daily hyphae growth rate at 20° C. In addition, HKHM25 had slower mycelial growth rate than ‘Hokuto 18gokin’ and ‘marmo22go’ at each temperature zone (Table 2).
  • Container An 850 polypropylene bottle (Capacity: 850 ml, diameter 58 mm) was used.
  • Culture medium Conifer sawdust, corn cob, rice bran and wheat bran were mixed at the dry weight ratio of 7:3:8:2, and the water content was adjusted to 65%.
  • the culture medium was filled up to the brim of the bottle at the rate of 540 ⁇ 20 g per bottle, and was sterilized at high pressure.
  • Starter culture About 20 ml of sawdust starter cultures per bottle was inoculated.
  • Culture Culture was performed at 22° C. for 50 to 90 days at 70% moisture.
  • Table 2 shows the characteristics of the ‘HKHM25’ and specific difference in characteristics as compared with the similar variety when culture was performed under the abovementioned conditions.

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Botany (AREA)
  • Developmental Biology & Embryology (AREA)
  • Environmental Sciences (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

The present variety of Bunashimeji mushroom named ‘HKHM25’ was cultivated by the gathering and repeated breeding of Bunashimeji mushrooms having thick and elastic stems, dark cap color, strong cap roll, and a large mushroom size, has a high quality and enhanced cultivation stability. This edible mushroom is exquisite in stability, reproducibility and uniformity when being produced.

Description

BACKGROUND OF THE INVENTION
This invention relates to a new and distinct variety of Bunashimeji mushroom, Hypsizygus marmoreus (Peck) H.E.Bigelow. This new variety named ‘HKHM25’, cultivated by repeated breeding of Bunashimeji mushroom having thick and elastic stems, dark cap color, strong cap roll, and a large mushroom size, has a high quality and enhanced cultivation stability, and ensures presentable stability, reproducibility, and uniformity.
The annual production of Bunashimeji mushrooms is approximately 120,000 tons in 2020, making it the second most consumed edible mushroom after enokitake mushroom in Japan. However, since Bunashimeji mushrooms have become available in large quantities in the market, Bunashimeji mushrooms with improved flavor, quality and shelf life has been pursued, which led to the development of ‘Hokuto 18gokin’ by our company. Further, our company has been conducting a wide variety of studies for the more stable cultivation and better quality of Bunashimeji mushrooms. Our company has developed a ‘marmo22go’ mushroom which is thicker, whiter, and has larger mushroom than ‘Hokuto 18gokin’.
Further, using ‘marmo22go’ as a parent, as a result of continuing breed improvement by cross breeding to improve stability and quality, ‘HKHM25’, which has thicker stems and stronger cap roll, as compared to ‘marmo22go’, was developed, confirming its stability, reproducibility, and uniformity.
SUMMARY OF THE INVENTION
The present invention is a new and distinct variety of mushroom plant characterized particularly by having thick and elastic stems, dark cap color, strong cap roll, and a large mushroom size, which can be cultivated by gathering and repeated breeding of fungal strains having a stability, reproducibility and uniformity when being produced. This novel and distinct variety of mushroom is identified as ‘HKHM25’.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 shows a phylogenetic tree illustrating the antecedents of ‘HKHM25’.
FIGS. 2A and 2B respectively show front and back images of a dual-culture of ‘HKHM25’.
FIGS. 3A and 3B respectively show front and back images of a dual-culture of ‘HKHM25’ and ‘Hokuto 18gokin’.
FIGS. 4A and 4B respectively show front and back images of a dual-culture of ‘HKHM25’ and ‘marmo22go’.
FIGS. 5A and 5B respectively show front and back images of a fungal flora of ‘HKHM25’ grown in a petri dish.
FIGS. 6A and 6B respectively show front and back image of a fungal flora of ‘Hokuto 18gokin’ grown in a petri dish.
FIGS. 7A and 7B respectively show front and back images of fungal flora of ‘marmo22go’ grown in a petri dish.
FIG. 8 shows an image of cultivation area of ‘HKHM25’.
FIG. 9 shows an image of cultivation area of ‘Hokuto 18gokin’.
FIG. 10 shows an image of cultivation area of ‘marmo22go’.
FIG. 11 shows an image of a fruit body of ‘HKHM25’.
FIG. 12 shows an image of a fruit body of ‘Hokuto 18gokin’.
FIG. 13 : shows an image of a fruit body of ‘marmo22go’.
FIG. 14 shows an image of density and surface color of fungal flora in cultured hyphae of ‘HKHM25’.
FIG. 15 shows an image of density and surface color of fungal flora in cultured hyphae of ‘Hokuto 18gokin’.
FIG. 16 shows an image of density and surface color of fungal flora in cultured hyphae of ‘marmo22go’.
FIG. 17 shows an image of development of aerial hypha of ‘HKHM25’.
FIG. 18 shows an image of development of aerial hypha of ‘Hokuto 18gokin’.
FIG. 19 shows an image of development of aerial hypha of ‘marmo22go’.
FIG. 20 shows an image of size and distribution of mottle of ‘HKHM25’.
FIG. 21 shows an image of size and distribution of mottle of ‘Hokuto 18gokin’.
FIG. 22 shows an image of size and distribution of mottle of ‘marmo22go’.
FIG. 23 shows an image of shape and thickness of stipe of ‘HKHM25’.
FIG. 24 shows an image of shape and thickness of stipe of ‘Hokuto 18gokin’.
FIG. 25 shows an image of shape and thickness of stipe of ‘marmo22go’.
 DETAILED DESCRIPTION OF THE INVENTION
‘HKHM25’ has been asexually reproduced by tissue culture, at HOKUTO CORPORATION in Japan. The history of the ‘HKHM25’ mushroom in terms of improvement period and the like are set forth in the following chronological list of each stage of variety improvement:
    • December 2014: Cultivation of ‘MH025615’ strain.
    • January 2017: Cultivation of ‘M11025617’ (‘marmo22go’) strain.
    • December 2019: ‘MH025615’ and ‘MH025617’ (‘marmo22go’) strains were crossed and a strain with thick stem, strong cap roll, and good quality among the obtained strains was selected as ‘MH025633’. We then repeated the cultivation test to distinguish the strains.
    • March 2021: After repeated cultivation tests, since distinguishability, stability, and uniformity were confirmed, the strain was named ‘HKHM25’ and its cultivation was completed. Applied for registration of a new variety to the Ministry of Agriculture, Forestry and Fisheries of Japan.
The above crossing is summarized in the phylogenetic tree illustrated in FIG. 1 .
The ‘HKHM25’ mushroom has the following characteristics: thick and elastic stems, dark cap color, strong cap roll, and overall large size.
(1) Comparison with Existing Variety by Dual Culture
Dual culture was performed for the ‘HKHM25’ mushroom and a similar variety to examine whether or not a zone line is formed.
Study Method:
As an examination method, a potato dextrose agar medium was used, and the ‘HKHM25’ mushroom and the similar variety were inoculated thereon face to face at an interval of 3 cm, and then culture was performed at 25° C. for 28 days to examine whether or not a zone line was formed.
Strains used for the comparison between present variety, ‘HKHM25’ and other varieties:
    • ‘HKHM25’: Present variety
    • ‘Hokuto 18gokin’: Variety similar to the present variety
    • ‘marmo22go’: Parent variety of the present variety
      Results:
Zone lines were formed between ‘HKHM25’ and all other co-cultured varieties
(TABLE 1, FIGS. 2 to 4)
This clearly shows that the present mushroom is a new variety.
Similar varieties
Hokuto 18gokin marmo22go
HKHM25 + +
+ is present and − is absent.
*Zone line formation was not observed in the dual culture between ‘HKHM25’ strains.

(2) Growth Characteristics of ‘HKHM25’
Study Method:
After inoculating an agar piece of the ‘HKHM25’ having a diameter of 5 mm and an agar piece of the similar variety having a diameter of 5 mm on a potato dextrose agar medium, preculture was performed at 25° C. for 4 days so as to make the regeneration of hyphae equal (about 10 mm in diameter), and then culture was performed for 7 days at intervals of 5° C. between 5° C. and 30° C. An average daily hyphae growth rate was calculated based on a hyphae growth rate for 7 days of the culture. Also, the optimum culture period for ‘HKHM25’ is 80 days, and the number of growing days at that time is 23.1 days.
Results:
‘HKHM25’ had the fastest average daily hyphae growth rate at 20° C. In addition, HKHM25 had slower mycelial growth rate than ‘Hokuto 18gokin’ and ‘marmo22go’ at each temperature zone (Table 2).
(3) Morphological Characteristics of the ‘HKHM25’ Mushroom in a Cultivation Example
Cultivation Method:
Container: An 850 polypropylene bottle (Capacity: 850 ml, diameter 58 mm) was used.
Culture medium: Conifer sawdust, corn cob, rice bran and wheat bran were mixed at the dry weight ratio of 7:3:8:2, and the water content was adjusted to 65%. The culture medium was filled up to the brim of the bottle at the rate of 540±20 g per bottle, and was sterilized at high pressure.
Starter culture: About 20 ml of sawdust starter cultures per bottle was inoculated.
Culture: Culture was performed at 22° C. for 50 to 90 days at 70% moisture.
Growth: After completing the culture, the inoculum is removed, and shifted to a growing room. Development was conducted under a temperature of 15±1° C., at humidity of 95% or about 2,000 ppm CO2 density. Also, light was not particularly irradiated until the first 14th day, then irradiated with about 500 to 1,000 Lx. The mushroom is harvested when the cap in the center of the stump has sufficiently opened.
Cultivation Results:
Table 2 shows the characteristics of the ‘HKHM25’ and specific difference in characteristics as compared with the similar variety when culture was performed under the abovementioned conditions.
In addition, according to R.H.S. Colour Chart, the color of the central part of cap: 199A, the color of the peripheral area of cap: 199C, the color of gill: 158C, the color of stripe: 155B. All color references are from The R.H.S. Colour Chart of The Royal Horticultural Society of London (R.H.S.), fifth edition published in 2007. Also, the descriptions are from mushrooms that are on the 24th day.
TABLE 2
Fungus characteristics Table of Hypsizygus marmoreus (Peck) H.E. Bigelow of Recording and Registration
(Please circle the applicable items for the characteristics.)
Characteristic
values
Remarks of similar
Characteristic values of present variety (comparison (Measured varieties
with standard varieties values, (Hokuto
Character 01 02 03 04 05 06 07 08 09 etc.) (marmo22go) 18gokin)
Physio-
logical
property
Dual
culture
Zone line none ob- — 09 09
formation served
Dislike- none ob- — 01 01
touch served
reaction
Density of cultured hyphae low medium
Figure USPP035511-20231128-P00001
 03 02 02
Develop- ment of aerial low medium
Figure USPP035511-20231128-P00002
 03 03 02
hypha
Color of surface of fungal flora white
Figure USPP035511-20231128-P00003
 pale yellow other 02 01 01
Color of back of fungal flora white
Figure USPP035511-20231128-P00004
 pale yellow other 02 02 02
Accommo-
dativeness
for
temperature
Optimal temperature for hyphal growth
Figure USPP035511-20231128-P00005
 24~ 26° C. 28~ 30° C. 03 21.9° C. 05 24.1° C. 04 22.5° C.
Hyphal growth rate 5° C./mm low
Figure USPP035511-20231128-P00006
 high 05 0.52 mm 06 0.70 mm 05 0.56 mm
Hyphal
growth
rate
10° C./mm low
Figure USPP035511-20231128-P00007
 medium 04 0.86 mm 05 1.16 mm 05 1.24 mm
15° C./mm low
Figure USPP035511-20231128-P00008
 high 05 1.65 mm 05 1.95 mm 05 1.80 mm
20° C./mm low
Figure USPP035511-20231128-P00009
 medium high 04 2.47 mm 05 2.79 mm 05 2.60 mm
25° C./mm low
Figure USPP035511-20231128-P00009
 medium high 04 2.01 mm 05 3.29 mm 04 2.47 mm
30° C./mm low
Figure USPP035511-20231128-P00009
 medium high 04 0.91 mm 07 2.17 mm 05 1.23 mm
Morpho-
logical
property
Cap
Cross sectional shape convex
Figure USPP035511-20231128-P00010
 flat con- cave other 02 02 02
Mottle of the surface few
Figure USPP035511-20231128-P00011
 many 02 02 02
Size of mottle small medium
Figure USPP035511-20231128-P00012
 03 01 03
Distribution of mottle
Figure USPP035511-20231128-P00013
 whole part 01 02 02
Clarity of mottle unclarity moderate
Figure USPP035511-20231128-P00014
 03 03 03
Size
Figure USPP035511-20231128-P00015
 medium large 03 21.2 mm 03 23.2 mm 02 20.4 mm
Color of central area white grey- yellow light grey- brown yellow- brown
Figure USPP035511-20231128-P00016
 dark tan dark grey tan dark grey brown other 05 199A 05 199C 05 199B
Color of peripheral area white grey- yellow light grey- brown yellow- brown
Figure USPP035511-20231128-P00017
 dark tan dark grey tan dark grey brown other 05 199C 05 199D 05 199D
Thickness thin medium thick
Figure USPP035511-20231128-P00009
 08 9.8 mm 09 11.3 mm 0.7 8.1 mm
Fleshy soft
Figure USPP035511-20231128-P00018
 hard 05 05 07
Gill
Color white
Figure USPP035511-20231128-P00019
 light orange yellow grey- yellow other 02 158C 02 158C 02 158C
Alignment normal
Figure USPP035511-20231128-P00020
 other 02 02 02
Width narrow
Figure USPP035511-20231128-P00021
 wide 02 02 02
Density low
Figure USPP035511-20231128-P00022
 high 02 02 02
Stipe
Shape long and thin short and thin long and thick short and thick
Figure USPP035511-20231128-P00023
 05 05 01
Length short
Figure USPP035511-20231128-P00024
 long 05 56.0 mm 05 53.9 mm 04 52.8 mm
Thickness thin medium thick
Figure USPP035511-20231128-P00009
 09 17.7 mm 08 16.5 mm 05 9.4 mm
Color
Figure USPP035511-20231128-P00025
 yellow- white grey grey- brown other 01 155B 01 155B 01 155B
Hair
Figure USPP035511-20231128-P00026
 few medium many 01 01 01
Fleshy soft medium
Figure USPP035511-20231128-P00027
 03 02 02
Ratio of maximum diameter of small medium
Figure USPP035511-20231128-P00028
 07 2.24 06 1.91 04 1.52
stem to
diameter
just below
cap
Cultural
property
Develop-
ment of
fruit body
Develop- ment group scattered
Figure USPP035511-20231128-P00029
 03 03 03
Optimal culture period short
Figure USPP035511-20231128-P00030
 long 05 80 days 05 80 days 06 90 days
Length of time from fruit body- short
Figure USPP035511-20231128-P00031
 long 05 23.3 days 04 21.9 days 05 22.3 days
formation
promotion
to fruit
body
harvesting
Optimal temperature for low medium
Figure USPP035511-20231128-P00032
 07 19° C. 07 19° C. 05 15° C.
primordial
development
Optimal temperature for fruit body low
Figure USPP035511-20231128-P00033
 high 05 15° C. 05 15° C. 05 15° C.
growth
Adaptivity for
illuminance
Adaptivity for
culture
Yield
Yield of fruit body few
Figure USPP035511-20231128-P00034
 many 05 125.4 g 06 137.0 g 02 98.4 g
The number of productive
Figure USPP035511-20231128-P00007
 few medium many 02 19.4 stems 03 21.7 stems 03 23.6 stems
stems
Other
property
Bitter none weak strong
ingredient
Disease
resistance
Ingredients
Notes:
Circle shows the characteristics of ‘HKHM25’.

The R.H.S. Colour Chart of The Royal Horticultural Society of London (R.H.S.), fifth edition published in 2007, The R.H.S. contact: R.H.S. Membership Department The Royal Horticultural Society PO Box 313 London, SW1P 2PE

Claims (1)

What is claimed is:
1. A new, distinct variety of Bunashimeji Mushroom as substantially illustrated and described in the specification.
US17/803,159 2022-03-08 2022-03-08 Bunashimeji mushroom named ‘HKHM25’ Active 2042-09-07 USPP35511P3 (en)

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