TWI411680B - Albino mutant of auricularia polytricha - Google Patents

Abstract

An albino mutant of Auricularia polytricha is disclosed. The fruit bodies of the albino mutant are grown in tuft, having opaquely ivory and double-flowered appearance, each of which is 3-9 cm in diameter and 1.0-2.0 mm thick. The fruit bodies of the albino mutant have white fine hairs on their back surface. The albino mutant provides a new edible fungus that is rich in nutrition, persists its crunchiness despite the long cooking processes and has good tasting.

Description

Edible Auricularia auricula

The present invention relates to a mutant strain of fungus, and more particularly to a mutant strain of Auricularia auricula.

The species of fungus that are widely cultivated and eaten internationally are Auricularia auricular-judae and Auricularia polytricha , which are edible fungi of the fungus of the fungus, and mainly in Taiwan. . At present, there are two main strains of hairy fungus planted and sold in Taiwan. Among them, the mushroom growers call it "black ear", which is generally based on fresh sales, while the other type is "red ear". Made for dry fungus. Early fungus was cultivated with segment wood, but the cost was high and the time to grow into fungus was too long. Nowadays, space bags are used to cultivate them on a case-by-case basis.

The fungus has a high nutritional value and is rich in polysaccharides of the fungus. It has the effects of lowering blood pressure and moistening the intestines after eating. However, although the black ear is rich in gum, it is not durable; the red ear is durable and has a crisp taste, but it has less gum.

Edible fungus can consume a variety of nutrients. The fungus is rich in protein, edible cellulose, vitamins, etc., and has high iron and calcium content, low calorie and only a very small amount of fat. Therefore, if you develop a new edible fungus with a good export, a variety of dishes, and a better appearance of the ingredients, you can provide consumers with more choices of ingredients.

The invention provides a mutant of Auricularia auricula, which is a new edible fungus, and the fruit body has an opaque beige color and is shaped like a petal of a peony flower, and has high nutritional value and good taste.

One aspect of the present invention provides a mutant of Auricularia auricula, deposited in the Food Industry Development Research Institute (FIRDI), No. BCRC 930154, characterized in that the plurality of fruiting bodies of the Auricularia auricularia aurantia mutant are clustered and fresh. An opaque beige, petal shaped like a peony flower, each of which has an average diameter of 3 cm to 9 cm, an average thickness of 1.0 mm to 2.0 mm, and a white fine fluff on the back of the ear. .

According to an embodiment of the present invention, the Auricularia auricula mutagenesis mutant has at least 29 free amino acids, including phosphoserine, taurine, aspartic acid, leucine, serine, and Tianmen Winter amide, glutamic acid, α-aminoadipate, glycine, alanine, α-amino-n-butyric acid, lysine, cystine, methionine, cysteine, different Aleucine, leucine, tyrosine, phenylalanine, beta-alanine, beta-aminoisobutyric acid, gamma-butylamine, ethanolamine, DL-isohydroxyisoamine, ornithine, amine Acid, histidine, arginine and valine.

According to an embodiment of the present embodiment, the content of the free amino acid in the auricular albino mutant is 0.1332% by weight.

The auricular albino mutant of the present invention is obtained by mutation and screening of the black wild strain of the black ear of the fungus, and is deposited in the Food Industry Development Research Institute (FIRDI) under the accession number BCRC 930154. The ribosomal deoxyribonucleic acid (rDNA) was sequenced by the Food Industry Development Institute to obtain a sequence fragment of sequence identification number 1 and sequence identification number 2. The sequence fragments of sequence identification number 1 and sequence identification number 2 were compared with the National Center for Biotechnology Information (NCBI) GenBank database, and sequence identification number 1 was similar to the sequence of Auricularia polytricha (HM448450). The degree is 99%, and the sequence similarity between sequence identification number 2 and Auricularia polytricha (AF261554) is 99%. The result of the alignment is that the mutant is a mycelium of Auricularia polytricha .

The mutation screening method, the bacteriological characteristics, and the culture method of the Auricularia auricula mutagen mutant of the present invention are described below.

1. Induction and screening methods for albino mutants:

The mutation screening method of the Auricularia auricular mutagenesis mutant of the present invention is described as follows. First, water (4500 mL), glucose (130 g), soy flour (90 g), corn flour (45 g), magnesium sulfate (2.3 g), potassium dihydrogen phosphate (4.5 g), and yeast powder (20.25 g) The bacterial culture solution was prepared and sterilized at a temperature of 121 ° C to 122 ° C for 1.5 hours at 1.5 atmospheres. The mycelium of the black fungus of the fungus cultured in potato dextrose agar (PDA) medium was cut into 12 cubes with a length of 2 mm to 3 mm and inoculated into 500 mL of the cells. In the culture medium, the culture was shaken for 9-10 days to form a hyphae. Then, the healthy hyphae are taken out and transferred to a new cell culture medium to regenerate the colonies.

In the process of shaking culture, the alternating temperature and carbon dioxide concentration can be used, for example, alternating 30 ° C or 6 ° C temperature, and / or with 6000 ppm or 500 ppm carbon dioxide concentration alternately, thereby stimulating albino whitening mutation The mycelium of the strain is formed.

The above steps are repeated a plurality of times, and the stimulation of alternating temperature and carbon dioxide concentration is added to continuously regenerate the aged hyphae and stimulate the formation of hyphae of the auricular albino mutant. After that, the liquid bacteria are planted into space capsules to produce fruiting bodies.

If the auricular albino mutant is not cultured, the number of times the culture liquid is regenerated is increased, and the aging and regenerated hyphae are forced to be mutated.

After the mutant of Auricularia auricula is cultured, the fruit body is separated, the mycelium is cultured, and the mycelial culture liquid is planted into a space bag for cultivation. Then, the white-haired albino mutants were selected, and the next stage of re-cultivation was carried out, and the re-cultivation was repeated repeatedly, so that the cultivated Auricularia auricula were all albino mutants.

Second, the morphological characteristics of colonies and fruiting bodies:

Please refer to FIGS. 1A to 1C, which are photographs showing the overall appearance, front and back photographs of the mycelium of the Auricularia auricula mutagen mutant according to an embodiment of the present invention on a potato dextrose agar slant medium.

The strain was inoculated on potato dextrose agar plate medium and malt extract agar (MEA) plate medium, and after 21 days of culture, the colony diameter was about 50 mm to 60 mm as shown in Figs. 1E to 1H.

Please refer to Figures 1E and 1F, which are photographs of the front and back of the cultured Auricularia auricular mutagenesis mutant on potato dextrose agar plate medium, respectively. Figure 1E shows that the colonies are white on the potato dextrose agar medium, and the aerial hyphae grow in a radial shape, with high and low fluctuations, and are cotton-like. After that, the aerial hyphae in the middle mature area were affixed to form an inner ring, and the outer edge of the colony was uniform. From the first F picture, the back surface of the colony was light yellow-brown, and the middle mature area was dark brown.

Please refer to Figures 1G and 1H, which are photographs of the front and back of the mycelium of the cultured Auricularia auricular amyloplast strain on the malt extract agar medium, respectively. In Fig. 1G, the colony was white on the periphery of the malt extract agar medium, and the inside was light beige. The aerial hyphae grew in a radial shape, which was cotton-like to villous, and then began from the middle of the mature mycelium. The volts are in a water-soaked ring with a uniform outer edge of the colony. The 1H figure is the back of the colony and is light brownish yellow.

Please refer to FIG. 1D, which is a button body structure of the mycelium of the Auricularia auricula mutagenesis mutant observed by an optical microscope, and the scale on the lower right side represents 2 μm. It can be seen from Fig. 1D that the mycelium has no sporulation structure, and the hyphae of the freshly cultured colonies are about (1-)2 to 3.5 (-5.5) μm wide, and the mycelial cell wall is thin and smooth.

Please refer to Fig. 2A, which is a photograph of the overall appearance of the fruiting body of the Auricularia auricula. Please refer to Fig. 2B together for another overall appearance photograph of the fruiting body of the Auricularia auricula. The fruiting bodies are clustered, opaque beige when fresh, and shaped like petals of peony flowers, each of which has an average diameter of 3 cm to 9 cm, and the average thickness of each fruiting body is 1.0 mm to 2.0 mm. The average outer diameter of the plexus is 12 cm to 22 cm, and the average height of the whole plexus is 6 cm to 8 cm. There are white fine fluff on the back of the ear. The hair is dense but short, and the small fluff is not visible to the naked eye. It needs to be observed with a magnifying glass.

Please refer to Figure 3, which is a mutant of Auricularia auricula (left) and ratio A photo of the overall appearance of a black wild strain (right side). Please refer to Fig. 4 together, which is a photograph of the overall appearance of the black wild strain of the comparative example. It can be seen from Figures 3 and 4 that the fruit bodies of the black wild strains are ear-shaped to disc-shaped, each of which has a larger diameter, and the villi are clearly visible at the back of the ear, and the villi are longer. The auricular albino mutant is composed of several small pieces of fruiting bodies, forming a petal like a peony flower.

Third, the analysis of nutrients:

Please refer to Table 1, which shows the results of component analysis of the Auricularia auricula mutagenesis mutant according to an embodiment of the present invention. As can be seen from the results of Table 1, the Auricularia auricula mutagenesis mutant has abundant nutrients including, for example, fat, protein, free amino acid, carbohydrate, polysaccharide, sodium and the like. In addition, the albino mutant of Auricularia auricula contains calcium, phosphorus and iron (results not shown).

Please refer to Table 2, which shows the types and contents of free amino acids contained in the Auricularia auricula mutagenesis mutant according to an embodiment of the present invention. Table 2 shows the compositional content of the free amino acid. From the results of Table 2, the Auricularia auricula mutagenesis strain contains at least 29 free amino acids, and contains 133.2 mg of free amino acid per 100 g of Auricularia auricula.

In addition, an acute toxicity test was conducted via the Food Industry Development Institute. The mice were fed a stomach tube for 2 weeks at a dose of 5 g/kg bw (animal body weight) of cooked Auricularia auricula (experimental group) or water (control group), and the clinical symptoms of the mice were observed. The results of the test showed that all mice had normal activity, no increase in body weight, and no mice died after 2 weeks of observation. After the end of the experiment, dissection was performed to visually observe the organs in each mouse body, which showed no obvious pathological changes in the organs of the mice, and no symptoms of poisoning were observed. From the above results, it was revealed that the oral acute albino mutant of the present invention has an oral acute toxicity LD ₅₀ value of more than 5 g/kg of animal body weight, which is not biotoxic, and thus can be applied to foods or other aspects.

Fourth, the training method:

In one embodiment, the above-mentioned obtained species of Auricularia auricula mutated mutant strain can further utilize a space bag to produce a fruit body (or a mushroom). Suitable nutrients for space bags include wood chips (89%), rice bran (3%), barley flour (3%), powder head (4%), and calcium carbonate (1%). Stir the nutrients evenly in the above proportions. . After that, water was added, and the mixture was uniformly stirred and then bagged, in which water accounted for 65% and the pH was adjusted to 7. Then, it was sterilized at a temperature of from 99 ° C to 101 ° C for 1.5 hours from 1.5 atm to 2.0 atm. After cooling, the above-mentioned obtained Auricularia auricularia aureus mutant strain is introduced into a sterile environment, and sealed and cultured in an environment of a temperature of 20 ° C to 25 ° C and a carbon dioxide concentration of 1000 ppm or less for 40 days to 45 days. After it is mature, open the space bag, increase the humidity to stimulate the mushroom, and harvest it at a temperature of 22 ° C to 26 ° C for 28 days.

It is worth mentioning that the above-mentioned nutrient ingredients of the present invention exclude the use of cottonseed hulls to cultivate fungus. The reason is that the cottonseed hull contained in the conventional nutrient component has the toxic chemical, gossypol, which may contaminate the albino mutant of Auricularia auricula. Consumption of cotton auricular whitening mutants containing cotton phenol can cause symptoms such as heart discomfort and male infertility.

5. Comparison with other fungus:

The auricular albino mutant of one embodiment of the present invention was compared with the existing black ear, white fungus, and black fungus of Auricularia auricula, and the results are shown in Table 3. The Auricularia auricula mutant has 29 free amino acids, and its yield and free amino acid species are much higher than the other three kinds of fungus.

Secondly, in terms of cultivation, the auricular albino mutant of the present invention is easy to cultivate, and the fruit body can be produced without symbiosis by two bacteria, wherein the first average yield of each space bag can be as high as 320 grams or more, and the economy The value is quite high. In contrast, Tremella needs to be mixed with two different strains before being transferred to space pack culture. However, when the above-mentioned strains are mixed, not only are they susceptible to contamination by other bacteria, but if the time of mixing the respective strains is different, it is impossible to symbiosis between the two bacteria and stimulate the production of fruiting bodies, thereby affecting the yield. As for the cultivation of snow fungus, although it is similar to the albino mutant, the production of snow fungus is less, and the first average yield of each space bag is only 120 grams or less.

It should be noted that, in evaluating the taste, the manner of cooking, and the appearance of the food, the amygdalin mutant of the present invention has more gelatin than the black ear, and has a crisp taste and no mushroom taste. Secondly, the auricular albino mutant of the present invention is more durable to cook, and the manner of cooking is more diverse, and the extraction, the juice, the hot pot, the stir-fry, the cold salad, and the like can be applied. Furthermore, the fruit body of the auricular albino mutant of the present invention is opaque beige, shaped like a petal of a peony flower, and has a more beautiful appearance, and can provide a new choice for edible fungus. In contrast, although the practicality of the black ear is also extensive, the black ear has a mushroom flavor, and many people are afraid to eat it. In addition, the black ear has a lot of glue but is not durable. Tremella and snow fungus have no mushroom flavor, but the white fungus mushroom umbrella is small like the petals of chrysanthemum, which is mostly used for extraction or dessert, while the snow fungus is mostly used for extraction or juice. Therefore, the mouthfeel, the manner of cooking, and the appearance of the food of the Auricularia auricula mutant of the present invention are far superior to the existing snow fungus, white fungus, and black ear.

The present invention has been disclosed in the above embodiments, and is not intended to limit the present invention. Any one of ordinary skill in the art to which the present invention pertains can make various changes without departing from the spirit and scope of the invention. The scope of protection of the present invention is therefore defined by the scope of the appended claims.

<110>Lin Hongzhang

<120> Edible Auricularia auricula

<160>2

<210>SEQ ID No: 1

<211>594

<212>DNA

<213> Auricularia polytricha

<220>

<223>rDNA ITS1-5.8S-ITS2

<400>1

<210>SEQ ID NO: 2

<211>917

<212>DNA

<213> Auricularia polytricha

<220>

<223>LSU rDNA D1-D3

<400>2

The above and other objects, features, advantages and embodiments of the present invention will become more <RTIgt; <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; A photograph of the overall appearance of potato dextrose agar slant medium.

Figure 1B is a positive view of the mycelium of the auricular albino mutant of Figure 1A. Face photo.

Fig. 1C is a photograph of the back side of the mycelium of the Auricularia auricula mutagenesis strain of Fig. 1A.

Fig. 1D is a view showing the structure of the mycelium of the mycelium of the Auricularia auricula mutagenesis observed by an optical microscope.

Figure 1E is a frontal photograph of the mycelium of the cultured Auricularia auricular mutagenesis strain on potato dextrose agar plate medium.

Fig. 1F is a photograph of the back side of the mycelium of the Auricularia auricula mutagenesis strain of Fig. 1E.

The 1G map is a frontal photograph of the mycelium of the cultured Auricularia auricula mutagenesis strain on the malt extract agar medium.

Figure 1H is a photograph of the back side of the mycelium of the Auricularia auricula mutagenesis strain of Figure 1G.

Figure 2A is a photograph of the overall appearance of the fruiting body of the Auricularia auricula.

Figure 2B is a photograph of another overall appearance of the fruiting body of the Auricularia auricula.

Fig. 3 is a photograph showing the overall appearance of a black fungus albino mutant (left side) and a black wild strain (right side) of a comparative example.

Fig. 4 is a photograph showing the overall appearance of a black wild strain of a comparative example.

Claims (2)

A mutant of Auricularia auricula, deposited in the Food Industry Development Research Institute (FIRDI), number BCRC 930154, characterized in that the plurality of fruiting bodies of the Auricularia auricula mutant are clustered, opaque beige when fresh, and shaped like a double flap a petal of peony, each of which has an average diameter of 3 cm to 9 cm, and an average thickness of each of the sub-entities of 1.0 mm to 2.0 mm and each of the sub-entities The surface of the back of the ear has white fine fluff, wherein the Auricularia auricula has at least 29 free amino acids, and the free amino acids comprise phosphorus serine, taurine, aspartic acid, lysine, silk Aminic acid, aspartame, glutamic acid, α-amino adipic acid, glycine, alanine, α-amino-n-butyric acid, proline, cystine, methionine, cystathionine Aminic acid, isoleucine, leucine, tyrosine, phenylalanine, beta-alanine, beta-aminoisobutyric acid, gamma-butylamine, ethanolamine, DL-isohydroxyisoamine, avian amine Acid, lysine, histidine, arginine and proline. The Auricularia auricular amyloplast mutant according to Item 1, wherein the free amino acid is 0.1332% by weight of the Auricularia auricula.