Strain of Pleurotus nebrodensis

Abstract

A novel strain of Pleurotus nebrodensis (Daewang No. 1, Accession No.: KACC93181P) and a method for cultivating it are provided. The novel strain of Pleurotus nebrodensis is different from the existing Pleurotus ferulae in shape and physiological characteristic, has an extra after-ripening period, can be grown at a low temperature of 22 to 25 C. and a low water content (RH) of 60 to 65%, can be cultivated in slightly acid environment of pH 5.5 to 6.5, can utilize bottle cultivation, and has a good shape not to be easily damaged in packaging. Thus, the novel strain of Pleurotus nebrodensis according to the present invention have good commercial value, are more resistant to environmental change, and can be mass produced by automation system and used for creating high value-added business in the food and agriculture industry.

Claims

1. A novel strain of Pleurotus nebrodensis designated Daewang No. 1, representative inoculum having been deposited at the Korean Agricultural Culture under Accession Number KACC93181P.

2. A method for incubating the novel strain of Pleurotus nebrodensis according to claim 1, the method comprising a step for incubating the Pleurotus nebrodensis in a medium comprising cotton seed husk of 30 to 50 vol %, corn powder of 2 to 8 vol %, wheat bran of 5 to 15 vol %, rice bran of 2 to 8 vol %, cotton seed meal of 2 to 8 vol %, corn cup of 10 to 26 vol %, sugar cane of 5 to 15 vol %, tangerine peel powder of 0.2 to 0.8 vol %, sugar of 0.2 to 0.8 vol %, soybean cake of 2 to 8 vol % and gypsum of 0.5 to 1.5 vol % (v/v %) based on total volume of the medium.

3. The method for incubating the novel strain of Pleurotus nebrodensis according to claim 2, wherein optimum temperature of strain growth is 22 to 25 C.

4. The method for incubating the novel strain of Pleurotus nebrodensis according to claim 2, wherein optimum humidity (RH) of strain growth is 60 to 65%.

5. The method for incubating the novel strain of Pleurotus nebrodensis according to claim 2, wherein optimum acidity (pH) of strain growth is 5.5 to 6.5.

6. The method for incubating the novel strain of Pleurotus nebrodensis according to claim 2, wherein after-ripening period is included in cultivation process.

7. The method for incubating the novel strain of Pleurotus nebrodensis according to claim 2, wherein the after-ripening period is processed at a temperature of 18 to 21 C.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIGS. 1 to 6 show cultivation pictures of a novel strain of Pleurotus nebrodensis according to the present invention.

DETAILED DESCRIPTION OF THE INVENTION

(2) The present invention provides a novel strain of Pleurotus nebrodensis, which is designated Daewang No. 1, representative having been deposited at the Korean Agricultural Culture under Accession Number KACC93181P.

(3) Further, the present invention provides a medium of the novel strain of Pleurotus nebrodensis.

(4) The present invention will now be described in detail.

(5) The novel strain of Pleurotus nebrodensis according to the present invention is characterized as being obtained by the following steps: isolating each of mono-spores from selected fruit bodies of Pleurotus ferulae and examining the toxicity of mononuclear strain; manufacturing mono-spore hybrid strains between each of strains; and selecting good quality of strains by cultivation and productivity examination.

(6) Pleurotus ferulae used in the present invention can be obtained by gathering or purchasing.

(7) In the present invention, the novel strain of Pleurotus nebrodensis can be obtained by the below described method.

(8) Original strains are obtained from fruit bodies of Pleurotus ferulae firstly gathered, and stabilization of medium is carried out with the productivity examination. Pleurotus ferulae secondly gathered is incubated in the stabilized medium, and the productivity examination is carried out. The most stable strains of them are selected. Mono-spores are isolated from fruit bodies formed by the selected strains and the feature of mononuclear strain is examined. And mono-spore hybrid strains between each of strains are manufactured and it is identified whether the manufactured mono-spore hybrid strains are hybridized. The hybridized strains are selected, artificially cultivated several times, and the productivity examination is carried out. And then by selecting the best one species of strains, the novel strain of Pleurotus nebrodensis according to the present invention can be obtained, which is excellent in productivity and stability.

(9) It is desirable that the medium comprises cotton seed husk of 30 to 50 vol %, corn powder of 2 to 8 vol %, wheat bran of 5 to 15 vol %, rice bran of 2 to 8 vol %, cotton seed meal of 2 to 8 vol %, corn cob of 10 to 26 vol %, sugar cane of 5 to 15 vol %, tangerine peel powder of 0.2 to 0.8 vol %, sugar of 0.2 to 0.8 vol %, soybean cake of 2 to 8 vol % and gypsum of 0.5 to 1.5 vol % (v/v %) based on total volume of the medium, but is not limited thereto.

(10) It is desirable to maintain the water content (RH) of the medium at 65%, but not limited thereto. Further, it is desirable to maintain the pH of the medium at 5.5 to 6.5, but is not limited thereto.

(11) It is desirable to incubate Pleurotus nebrodensis in the medium, but is not limited thereto.

(12) It is desirable that bottle cultivation is used to cultivate Pleurotus nebrodensis, but is not limited thereto.

(13) It is desirable that the optimum growth temperature of the strain of Pleurotus nebrodensis is 22 to 25 C., but is not limited thereto.

(14) It is desirable that the optimum growth humidity (RH) of strain of Pleurotus nebrodensis is 60 to 65%, but is not limited thereto.

(15) It is desirable that the optimum growth acidity (pH) of strain of Pleurotus nebrodensis is 5.5 to 6.5, but is not limited thereto.

(16) It is desirable that after-ripening period is included in cultivation process of Pleurotus nebrodensis, but is not limited thereto.

(17) The novel strain of Pleurotus nebrodensis according to the present invention is different from the existing Pleurotus ferulae in shape and physiological characteristic, has an extra after-ripening period, can be grown at a low temperature of 22 to 25 C. and a low water content (RH) of 60 to 650, can be cultivated in slightly acid environment of pH 5.5 to 6.5, can utilize bottle cultivation, and has a good shape not to be easily damaged in packaging. Thus, the novel strain of Pleurotus nebrodensis according to the present invention have good commercial value, are more resistant to environmental change, and can be mass produced by automation system. In conclusion, the present invention could be used for creating high value-added business in the food and agriculture industry.

(18) The present invention provides a method for cultivating a novel strain of Pleurotus nebrodensis.

(19) It is desirable that the cultivating method comprises the steps as follows:

(20) 1) manufacturing a medium comprising cotton seed husk of 30 to 50 vol %, corn powder of 2 to 8 vol %, wheat bran of 5 to 15 vol %, rice bran of 2 to 8 vol %, cotton seed meal of 2 to 8 Vol %, corn cob of 10 to 26 vol %, sugar cane of 5 to 15 vol %, tangerine peel powder of 0.2 to 0.8 vol %, sugar of 0.2 to 0.8 vol %, soybean cake of 2 to 8 vol % and gypsum of 0.5 to 1.5 vol % based on total volume of the medium;

(21) 2) putting the medium in bottles and sterilizing by high pressure;

(22) 3) inoculating the sterilized medium with seed of Pleurotus nebrodensis and incubating the seed;

(23) 4) thinning out a part of Pleurotus nebrodensis produced in the incubated medium, and incubating the rest during an after-ripening period; and

(24) 5) growing up the incubated Pleurotus nebrodensis.

(25) The cultivating method is not limited to the above described steps but may include alternative or additional steps if necessary.

(26) The cultivating method of the novel strain of Pleurotus nebrodensis can be industrialized and bottle-cultivated. Further, because little manpower and small space are required for the method, it can be cultivated by automation system and mass produced.

(27) The present invention also provides a fruit body of the novel strain of Pleurotus nebrodensis.

(28) Exemplary embodiments of the present invention will now be described for understanding of the present invention. The following embodiments provide for understanding of the present invention and are not intended to limit a technical scope of the present invention.

Embodiment 1: Manufacturing of a Novel Strain of Pleurotus nebrodensis

(29) In order to manufacture the novel strain of Pleurotus nebrodensis according to the present invention, we have visited Xinjiang Province, China, and firstly gathered wild Pleurotus ferulae. After returning to Korea, we obtained original strain from the fruit body of the gathered Pleurotus ferulae, and tried to cultivate the obtained original strain artificially.

(30) The original strain is incubated in a medium which comprises cotton seed husk of 30 to 50 vol %, corn powder of 2 to 8 vol %, wheat bran of 5 to 15 vol %, rice bran of 2 to 8 vol %, cotton seed meal of 2 to 8 vol %, corn cob of 10 to 26 vol %, sugar cane of 5 to 15 vol %, tangerine peel powder of 0.2 to 0.8 vol %, sugar of 0.2 to 0.8 vol %, soybean cake of 2 to 8 vol % and gypsum of 0.5 to 1.5 vol % based on total volume of the medium. And the stabilization of the medium was carried out with repetitive productivity examination.

(31) In order to breed a strain different from the existing species, Pleurotus ferulae was gathered secondly, the gathered Pleurotus ferulae was incubated in the stabilized medium, and the productivity examination was carried out. Three strains were gathered, two of them stable for cultivating were selected, and mono-spores were isolated from fruit bodies formed by the selected two strains, respectively.

(32) Using the isolated mono-spores, the feature of mononuclear strain was examined, and total 100 mono-spore hybrid strains between each of strains were manufactured. The clamp connection of the manufactured mono-spore hybrid strains was observed with a microscope to identify whether the manufactured mono-spore hybrid strains are hybridized. The mono-spore hybridization was tried for total 100 strains, 73 strains of them, in which hybridization was identified, were selected. The selected 73 strains were cultivated several times by a bag cultivation method, and the productivity examination was carried out. And then best strains were selected by investigating shape of pileus, color and gloss of pileus, thickness of stipe, length of stipe and so on of the fruit body grown in the cultivation test. The selected strains have a short duration of incubation and cultivation, are large in number, have an umbrella shaped pileus, and have a fleshy body dense rather than hard. The productivity examination was carried out for selected 73 strains from the mono-spore hybridization described in the above, and the best one strain was selected. As a result, a novel strain designated Daewang No. 1 is a selected interspecific hybrid between Pleurotus ferulae and Pleurotus nerbrodensis.

(33) The present inventors confirmed the obtained novel strain as a novel strain of Pleurotus nebrodensis according to the present invention, and deposited it to the RDA-Genebank Information Center of the Korean Academy of Agricultural Science on Apr. 22, 2013 (Accession name: Daewang No. 1, Accession No.: KACC93181P)

Embodiment 2: Cultivation Conditions of a Novel Strain of Pleurotus nebrodensis

(34) (a) Manufacturing of Medium

(35) A medium of a novel strain of Pleurotus nebrodensis was manufactured, which comprises cotton seed husk of 40 vol %, corn powder of 5 vol %, wheat bran of 10 vol %, rice bran of 5 vol %, cotton seed meal of 5 vol %, corn cob of 18 vol %, sugar cane of 10 vol %, tangerine peel powder of 0.5 vol %, sugar of 0.5 vol %, soybean cake of 5 vol % and gypsum of 1 vol % based on total volume of the medium.

(36) The water content (RH) of the medium was maintained at 65%, and the pH was maintained at 5.5 to 6.5.

(37) (b) Preparing of Incubation Container

(38) Incubation containers of 1100 cc75, 1300 cc80 and 1400 cc85 were prepared for bottle cultivation.

(39) (c) Sterilizing of Medium and Incubation Container

(40) The medium manufactured in (a) were put in the containers prepared in (b). And then the containers filled with the medium were high-pressure sterilized at 105 C. and 0.2 kgf/cm.sup.2 for 3 hours, and cooled at room temperature.

(41) (d) Incubating Condition

(42) The medium cooled in (c) was inoculated at 22 to 25 C. in the darkroom for 25 days, and an after-ripening process was carried out at 18 to 21 C. for 20 day.

(43) (e) Growth Condition of Fruit Body

(44) The fruit bodies formed in (d) were grown up under the condition of a temperature of 13 to 15 C., a relative humidity (RH) of 85 to 95% and a density of carbon dioxide of below 1,000 ppm, for 20 to 25 days.

Comparative Example: Obtaining and Cultivation Condition of Pleurotus ferulae

(45) 1. Obtaining of Pleurotus ferulae

(46) Pleurotus ferulae (or A-Wie-Go) firstly gathered in Embodiment 1 was used as a comparison group. The Pleurotus ferulae not bred or hybridized was used as it is, without further processing.

(47) 2. Cultivation Conditions of Pleurotus ferulae

(48) (a) Manufacturing of Medium

(49) (1) Medium A

(50) Medium A of Pleurotus ferulae was manufactured, which comprises sawdust of 24.09 vol %, wheat bran of 6.02 vol %, cotton seed meal of 6.02 vol %, corn cob of 9.03 vol %, rice bran of 3.01 vol %, soybean cake of 3.01 vol %, sugar cane of 7.83 vol %, tangerine peel powder of 0.6 vol %, lime hydrate of 0.6 vol %, red candy (Hong tang) of 0.6 vol % and water of 65.0 vol % (v/v %) based on total volume of the medium.

(51) (2) Medium B

(52) Medium B of Pleurotus ferulae was manufactured, which comprises cotton seed husk of 30.0 vol %, cotton seed meal of 6.06 vol %, wheat bran of 7.27 vol %, sugar of 0.6 vol %, soybean cake of 3.03 vol %, corn cob of 12.12 vol %, lime of 0.6 vol %, tangerine peel powder of 0.6 vol % and water of 39.39 vol % (v/v %) based on total volume of the medium.

(53) (3) Water Content and Acidity Control

(54) The water content (RH) of mediums A and B manufactured in (1) and (2) was maintained at 60 to 70%, and the pH was maintained at 7.5 to 8.5.

(55) (b) Preparing of Incubation Container

(56) Incubation containers of 1100 cc75, 1300 cc80 and 1400 cc85 were prepared for bottle cultivation.

(57) (c) Sterilizing of Medium and Incubation Container

(58) The medium A and B manufactured in (a) were put in the containers prepared in (b). And then the containers filled with the medium were high-pressure sterilized at 105 C. and 0.2 kgf/cm.sup.2 for 3 to 4 hours, and cooled at room temperature.

(59) (d) Incubating Condition

(60) The medium cooled in (c) was incubated at 22 to 25 C. in the darkroom for 25 to 30 days.

(61) (e) Growth Condition of Fruit Body

(62) The fruit bodies formed in (d) were grown up under condition of a temperature of 13 to 18 C., a relative humidity (RH) of 85 to 95% and a density of carbon dioxide below 1,000 ppm, for 20 to 25 days.

Embodiment 3: Incubating and Cultivating of a Novel Strain of Pleurotus nebrodensis

(63) In order to incubate and cultivate a novel strain of Pleurotus nebrodensis according to the present invention, the following described processes were carried out.

(64) In order to manufacture the medium of the novel strain of Pleurotus nebrodensis described in (a) of Embodiment 2, all base materials of the medium described above were mixed by a mixer (SG-4600D) for 40 minutes. Underground water was added to the mixture and mixed for 60 to 90 minutes so that water content of the medium was adjusted at a humidity of 60 to 650, and then the medium was put in bottles. The bottles filled with the medium was sterilized at high temperature (by a dry sterilizer 14-LMC or a high pressure sterilizer SHW-M110), and then cooled. The seed cut off oxygen by sealing the bottles was parceled out to a growth chamber, and laid and stacked on the frames installed in the growth chamber. It is desirable that the bottles are stacked up in three stages and the frames are installed in four or five stages. After adopting the seed, the growth chamber was maintained at a temperature of 15 to 18 C. and was observed for 7 to 10 days without opening the bottles. After adopting, the growth chamber was not humidified, the density of carbon dioxide of the chamber was maintained below 1,000 ppm, and light of 1000 to 1500 lux was supplied to the chamber for 10 to 12 hours per day. Hyphae have formed again, and the mushroom has begun to germinate. When fruit bodies of the mushroom in the bottles were grown up to the size of a soybean, the bottles are opened. After opening the bottles, when mushrooms were grown up to the size of a corn ear, only one mushroom grown up to the fullest was reserved, but the rest were thinned out. And then, for after-ripening, in the growth chamber, the humidity was maintained at 85 to 900, the temperature was maintained at 13 to 15 C. (by using a constant-temperature oven C-1B2, 3, 4), a density of carbon dioxide was adjusted below 700 ppm, and light of 1000 to 1500 lux was supplied for 10 to 12 hours per day. The mushrooms were harvested on 10th to 12th day from germination, and the growth chamber was not humidified on 1 day before harvest time. Of the obtained mushrooms, the mushrooms of 150 to 170 g had highest quality.

Embodiment 4: Comparison of Shape Characteristics Between a Novel Strain of Pleurotus nebrodensis and of Pleurotus ferulae

(65) In order to compare shape characteristics between the novel strain of Pleurotus nebrodensis according to the present invention and Pleurotus ferulae of the comparison group, the shape analysis was carried out for the novel strain of Pleurotus nebrodensis manufactured in Embodiment 1, Pleurotus ferulae prepared in Comparative Example and the fruit bodies of Pleurotus nebrodensis obtained Embodiment 3.

(66) 1. Pileus

(67) The pileus of the novel strain of Pleurotus nebrodensis had high white color similar to the comparison group. Further, the pileus had globular shape and its length was measured to be 10 to 15 cm. On the other hand, the pileus of Pleurotus ferulae of the comparison group had semi-spherical shape and its length was measured to be 6 to 13 cm.

(68) 2. Stipe

(69) The stipe of the novel strain of Pleurotus nebrodensis was thick and short comparatively, and its length is measured to be 2 to 3 cm. Further, the width of the stipe of the novel strain of Pleurotus nebrodensis was measured to be 2 to 3 cm so that the ratio of length to width is identified to be 1:1.

(70) On the other hand, the stipe of Pleurotus ferulae of the comparison group was thick and long, and its length is measured to be 3 to 8 cm. Further, the width of the stipe of Pleurotus ferulae was measured to be 4 to 6 cm so that the ratio of length to width is identified to be 3:1.

(71) 3. Fruit Body

(72) The novel strain of Pleurotus nebrodensis has a structure in which fruit body includes the stipe, and the shape of the fruit body is globular. Further, the fruit body of the novel strain of Pleurotus nebrodensis was ontogeny type the same as that of Pleurotus ferulae of the comparison group, but its shape was more stable relatively.

Embodiment 5: Comparison of Physiological Characteristics Between a Novel Strain of Pleurotus nebrodensis and of Pleurotus ferulae

(73) In order to compare physiological characteristics between the novel strain of Pleurotus nebrodensis according to the present invention and Pleurotus ferulae of the comparison group, optimum temperature, optimum water content and optimum acidity of strain growth were compared between the novel strain of Pleurotus nebrodensis manufactured in Embodiment 1 and Pleurotus ferulae prepared in Comparative Example.

(74) 1. Optimum Temperature of Strain Growth

(75) It was identified that the optimum temperature of strain growth of the novel strain of Pleurotus nebrodensis is 22 to 25 C., and the optimum temperature in the after-ripening period is 18 to 21 C. On the other hand, it was identified that the optimum temperature of strain growth of Pleurotus ferulae is 25 to 28 C., and there are not an extra after-ripening period, unlike the novel strain of Pleurotus nebrodensis according to the present invention.

(76) Therefore, it was identified that the novel strain of Pleurotus nebrodensis according to the present invention can be grown at lower temperature than Pleurotus ferulae of the comparison group and has an extra after-ripening period.

(77) 2. Optimum Water Content of Strain Growth

(78) It was identified that the optimum water content (RH, %) of strain growth of the novel strain of Pleurotus nebrodensis is 60 to 650. On the other hand, it was identified that the optimum water content of strain growth of Pleurotus ferulae is 65 to 70%.

(79) Therefore, it was identified that the novel strain of Pleurotus nebrodensis according to the present invention can be grown at lower water content than Pleurotus ferulae of the comparison group.

(80) 3. Optimum Acidity of Strain Growth

(81) It was identified that the optimum acidity (pH) of strain growth of the novel strain of Pleurotus nebrodensis is pH 5.5 to 6.5. On the other hand, it was identified that the optimum acidity of strain growth of Pleurotus ferulae is pH 7.5 to 8.5.

(82) Therefore, it was identified that the novel strain of Pleurotus nebrodensis according to the present invention can be grown in slightly acid condition.

(83) While the invention has been shown and described with reference to a certain preferred embodiment thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

(84) [Accession Number]

(85) Depositary authority: RDA-Genebank Information Center of the Korean Academy of Agricultural Science

(86) Accession No.: KACC93181P

(87) Accession Date: 20130422