CN104145709A - Black fungus strain on plateau - Google Patents

Abstract

A plateau black fungus strain, which relates to a black fungus strain. It is black fungus HDXZ-1 (Auricularia auricular HDXZ-1), which is preserved in China Center for Type Culture Collection with the preservation number CCTCC NO: M2013032. The invention is applied to the field of black fungus cultivation.

Description

A plateau black fungus strain

technical field

The invention relates to a black fungus strain.

Background technique

Black fungus is a saprotrophic edible fungus. In the two stages of growth and development, mycelium penetrates into the substrate, and the organic matter in the degraded substrate absorbs the released nutrients to provide its own nutrition and reproductive needs. Black fungus is a traditional edible fungus in my country. It is known as "vegetable meat and meat among vegetarians". It is known as a human body scavenger and is a healthy food in people's lives.

Tibetan black fungus is the best among mountain delicacies on the Qinghai-Tibet Plateau. It is a saprophytic fungus that grows on rotten wood and consists of mycelium and fruiting body. The unique environmental conditions of Tibet endow plateau black fungus with the characteristics of high nutritional value and high price; but also because of the special geographical environment of Tibet, such as high ultraviolet light and different forest matrix environment, the yield of local fungus on the plateau is generally low, which also makes the inland excellent The variety of fungus is "acclimatized" in Tibet, and its quality and taste have declined significantly.

Contents of the invention

The invention provides a black fungus plant suitable for planting under plateau environmental conditions, which has high planting yield and good commercial performance in the plateau area, and its comprehensive performance is better than that of inland transplanted varieties.

The black fungus HDXZ-1 (Auricularia auricular HDXZ-1) of the present invention is preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC NO: M2013032.

Plant black fungus HDXZ-1 (Auricularia auricular HDXZ-1) of the present invention in Tibet, and the yield dry weight of each fungus bag (the size of the fungus bag is Φ16.5cm×33cm, and the inoculation block is 1cm×2cm×2cm) reaches 48.7±0.26 g, the yield is basically the same as that of Hei 29 (dry weight per mushroom bag: 50.3±0.29g) and Yuanbao 2 (dry weight per mushroom bag: 49.7±0.31g) planted in the inland plain area (Dongning, Heilongjiang) Quite equivalent, 1.2 times that of Hei 29 transplanted to Tibet (40.6±0.31g dry weight per mushroom bag), 1.1 times that of Yuanbao 2 (45.7±0.28g dry weight per mushroom bag), It has the advantage of high output.

The black fungus HDXZ-1 (Auricularia auricular HDXZ-1) of the present invention has the characteristics of good taste and better than the fungus transplanted in the interior.

The cultivation season of the black fungus HDXZ-1 (Auricularia auricular HDXZ-1) of the present invention does not conflict with the traditional busy farming time, so that the farmers and herdsmen have something to do and money to make during the slack time. The use of bags for cultivation does not occupy arable land, and can be cultivated on tidal flats and gravel land, which can effectively expand the land use area. The income of farmers and herdsmen in plateau areas can be increased by cultivating black fungus HDXZ-1 (Auricularia auricularHDXZ-1). 200,000 bags need 20 mu of land, and it is estimated that about 19,000 catties of dried ears can be harvested. According to the raw material cost of each bag is 1.2 yuan , the labor cost is 1 yuan, and the price is 150 yuan per catty, which can bring a net income of 2.41 million yuan, which is beneficial to the local economic development.

Black fungus HDXZ-1 (Auricularia auricular HDXZ-1) is Auricularia auricula, which belongs to the genus Auricularia (Auricularia); it is preserved in the China Center for Type Culture Collection (CCTCC), the preservation address is Wuhan University, and the preservation date is January 2013 On March 21, the deposit number is CCTCC NO: M2013032.

Description of drawings

Figure 1 is a phylogenetic tree constructed by selecting fungi with more than 99% homology of black fungus HDXZ-1 (Auricularia auricular HDXZ-1).

Fig. 2 is a graph showing the result of hardness contrast test in Embodiment 2.

Fig. 3 is a graph showing the results of an elastic contrast test in Embodiment 2.

Detailed ways

The technical solution of the present invention is not limited to the specific embodiments listed below, but also includes any combination of the specific embodiments.

Specific embodiment 1: In this embodiment, black fungus HDXZ-1 (Auricularia auricular HDXZ-1) was preserved in the China Center for Type Culture Collection, the preservation date is January 21, 2013, and the preservation number is CCTCC NO: M2013032.

Black fungus HDXZ-1 (Auricularia auricular HDXZ-1) was collected from Nanyigou Scenic Spot, Milin County, Tibet, at an altitude of 2800m.

Black fungus HDXZ-1 (Auricularia auricular HDXZ-1) was initially identified as black fungus by morphology.

The collected black fungus HDXZ-1 (Auricularia auricular HDXZ-1) specimens were removed from the dirt, fumigated with potassium permanganate-formaldehyde solution for 30 minutes, and then the fruiting bodies were placed on the ultra-clean bench for aseptic separation ( Pick up the gelatinous part of the fruiting body with an inoculation shovel), inoculate the separated gelatinous part (soybean grain size) into the center of the PDA medium, cultivate it in a constant temperature incubator at 25°C for 15 days, and then pick the tip mycelium at the edge of the colony , and transplanted to the test tube medium (1L test tube medium consists of 200g peeled potato, 20.0g glucose, 3.0g KH ₂ PO ₄ , 1.5g MgSO ₄ , 14.0g agar, 75g wheat bran and the rest of the water) in the center of the slant Cultivate at 25°C until the hyphae cover the slope, and repeat this 3 times. The purified strains were inserted into a sawdust medium (the sawdust medium was composed of 40% dry material and 60% water, and the dry material was composed of 78% broad-leaved hardwood sawdust, 20% Wheat bran or rice bran, 1% gypsum and 1% white sugar) are preserved in test tubes.

Antagonism test:

The black fungus HDXZ-1 (Auricularia auricular HDXZ-1) was collected with the control fungus strains (Auricularia auricularia No. 29, Yuanbao No. 2 and another black fungus strain collected in Bomi County, Tibet Tibet No. 8 - Tibet No. 8 From Bomi County, Tibet, at an altitude of 2900m, the base material of the fungus is birch, collected by Zhang Yanlong, the collection time is 2010, and it is now preserved in the 211 Laboratory of Heilongjiang University, and released to the public), and the independence of the strains is tested in pairs. , were respectively inoculated to both ends of the test tube containing PDA medium, and each combination was repeated three times. After inoculation with the marked serial number, incubate at a constant temperature at 25°C, observe once every 12 hours, find and discard test tubes contaminated by miscellaneous bacteria in time, and replace the slant of a new test tube for inoculation and culture again. After contacting the mycelia of the strains in the tube, observe and record the contact parts of the mycelium, the degree of affinity of the mycelium and the formation of antagonistic lines. The specific morphological characteristics of uplift, isolation type, and groove type refer to the provisions of "GB/T12728 Terminology of Edible Fungi" Judgment: "-" means affinity, "+" means incompatibility but no obvious antagonistic line, "++" means incompatible and obvious antagonistic line. The results of the antagonism test are shown in Table 1.

Table 1 Antagonism results

strain Black 29 Yuan Bao No. 2 Tibet 8 Black fungus HDXZ-1 + + ++

Comparison test of mycelium growth and growth rate (the test was carried out in Tibet):

Black fungus HDXZ-1 (Auricularia auricular HDXZ-1) was inoculated according to the ratio of first-level strains: second-level strains: third-level strains = 1:7:560 (one first-level strain was inoculated with seven second-level cultures Base, 1 secondary strain inoculates 80 tertiary medium). Each strain of the first-grade strains is connected with 5 test tubes (PDA medium for the first-grade strains), and the second-grade strains (the medium for the second-grade strains is composed of 40% dry material and 60% water by mass percentage, The dry material is composed of 78% hardwood sawdust, 20% wheat bran or rice bran, 1% gypsum and 1% white sugar by mass percentage) and tertiary strains (the tertiary strain medium is based on mass The percentage is composed of 38% dry material and 62% water, and the dry material is composed of 76% Qinggang sawdust, 22% highland barley bran, 1% gypsum and 1% lime by mass percentage) to inoculate 100 bags and 300 bags respectively bag, set 3 repetitions.

(1) First inoculate black fungus HDXZ-1 (Auricularia auricular HDXZ-1) in the center of the slope of the PDA test tube in the ultra-clean workbench, place it in a constant temperature incubator at 25°C, and observe the growth of black fungus HDXZ-1 at room temperature Conditions, such as mycelium thick, thick degree, color and so on.

(2) Inoculate the black fungus HDXZ-1 with secondary bacteria in the inoculation box, and then put the secondary bacteria into the cultivation room sterilized by the bacteria room disinfection king in advance (place the inoculation port upright), first Cultivate at 29°C, and change to 25°C after the strains eat the material. After the mycelia cover the entire surface of the material, observe the growth of the mycelium of each strain, such as the thickness, density, and color of the mycelium.

(3) Inoculate the third-grade bacteria of black fungus HDXZ-1 on the inoculation table according to the aseptic operation procedure (the size of the inoculation block is 1cm×2cm×2cm), and then place the third-grade bacteria horizontally on the culture rack (inoculation Leave an appropriate gap at the mouth) to cultivate in an environment of 20°C, and open the door for ventilation for 1 hour every day when there is no wind. When the mycelium is covered with the material surface, randomly select 10 bags, mark the length of the fungus every 2 days, observe the growth of the mycelium at the same time, measure its length and record the full time, calculate the daily average mycelial growth rate, and Perform analysis of variance.

Table 2 shows the growth of black fungus HDXZ-1 (Auricularia auricular HDXZ-1) at all levels.

Table 2 The growth of strains at all levels

It shows that the mycelium of black fungus HDXZ-1 (Auricularia auricular HDXZ-1) grows vigorously, the mycelium is strong, white and grows well.

Table 3 The growth rate (mm) on the tertiary bacteria medium

Plateau cultivation ear test:

In the Linzhi area of Tibet, the plateau cultivation ear experiment was carried out. The cultivation medium (bacteria bag) was mixed with water and dry material. The mass ratio of water and dry material was 38:62. The dry material was composed of 76% Qinggang sawdust, Composition of 22% highland barley bran, 1% gypsum and 1% lime; the temperature during the cultivation of fungus bags is 20°C; the sterilization time of the cultivation medium is 9 hours in a simple sterilizer in the plateau area; choose 4 In the first ten days of the month, the fungus bags were cultivated and managed in the ground; in the field management stage of black fungus, the best cut shape was "∣" shape, and the best cut number was 140.

The black fungus HDXZ-1 (Auricularia auricular HDXZ-1) covered with fungus bags was taken to the ground for management, the ear-out situation of the strains and the contamination by miscellaneous bacteria were observed, and the commercial performance of the fruiting body was compared and analyzed after the ears were harvested. The bag records the yield of fruiting bodies.

Table 4 strain production situation

Table 5 Performance of plateau cultivated commodities

strain fruiting body color Flower type opening situation Taste Black fungus HDXZ-1 Ventral dark brown dorsally Bowl open piece, small root Soft and slippery

The commercial performance of black fungus HDXZ-1 meets the requirements of excellent black fungus products.

Submit the ITS sequence of black fungus HDXZ-1 (Auricularia auricular HDXZ-1) strain to NCBI, use the BLAST program to compare the homology with the sequence library already included in GenBank, download MEGA5.0 and select the homology above 99% According to the phylogenetic tree (as shown in Figure 1), the Auricularia auricularia HDXZ-1 (Auricularia auricular HDXZ-1) strain is in the same branch as the Auricularia auricularia strain 428-based subgroup in the database From this, it can be determined that black fungus HDXZ-1 (Auricularia auricular HDXZ-1) is a species of black fungus in the genus Auricularia.

The differences between black fungus HDXZ-1 (Auricularia auricular HDXZ-1) and black fungus strain 428 are shown in Table 6.

Table 6

strain Black fungus HDXZ-1 Black fungus strain 428 mycelial germination temperature 18°C 21°C optimum growth pH 7.5 7.3 optimum growth temperature 25℃-28℃ 23°C-26°C Optimum water content for growth 60%-62% 62%-65% Product performance of sub-entities small root bowl Single piece of small root Yield of pouches 48g 47g

Black fungus HDXZ-1 (Auricularia auricular HDXZ-1) was rehydrated in a water bath for 60 minutes, and the surface moisture was blotted with absorbent paper, and the rehydration rate was 90%. Black fungus HDXZ-1 (Auricularia auricular HDXZ-1) before rehydration, the ear piece is small and broken, dark and dull in color; after rehydration, the ear piece is thinner, dark brown and dull, with better elasticity and smoothness The sex is average, the overall taste is good.

Specific implementation mode 2: In this implementation mode, a comparative test is carried out on black fungus HDXZ-1 (Auricularia auricular HDXZ-1), outstanding auricularia varieties from the mainland, and excellent auricularia varieties transplanted to Tibet:

Sample No. 1 is black fungus in Dongning, Heilongjiang, which is black fungus Yuanbao No. 2 isolated from Bidekin Edible Fungi Research Institute in Dongning County;

Sample No. 2 is black fungus in Dongning, Heilongjiang, which is black fungus A4 isolated from Bidekin Edible Fungi Research Institute in Dongning County;

Sample No. 3 is Yuanbao black fungus No. 2 transplanted to Tibet;

Sample No. 4 is black fungus A4 transplanted to Tibet;

Sample No. 5 is black fungus HDXZ-1 (Auricularia auricular HDXZ-1).

1. The hardness comparison test results are shown in Figure 2, which shows that the hardness of the inland fungus transplanted to Tibet has increased significantly, and is significantly larger than that of black fungus HDXZ-1 (Auricularia auricular HDXZ-1), and the taste has decreased significantly.

2. The results of the elasticity comparison test are shown in Figure 3, which shows that the elasticity of the inland fungus transplanted to Tibet has decreased significantly, and is also lower than that of black fungus HDXZ-1 (Auricularia auricular HDXZ-1), and the taste is poor.

3. The test results of the macronutrient components of black fungus are shown in Table 7. Black fungus HDXZ-1 (Auricularia auricularHDXZ-1), that is, sample No. 5, has higher water and protein content than the other 4 samples, crude fat, reducing sugar and total sugar The content was significantly lower than that of the other 4 samples, and black fungus HDXZ-1 (Auricularia auricular HDXZ-1) was more in line with the current diet concept of high protein, high dietary fiber and low fat intake.

Table 7 The list of macronutrients in black fungus samples (unit: g/100g)

1 2 3 4 5 moisture 8.29±0.05b 8.49±0.03b 8.34±0.04b 8.02±0.07b 9.36±0.1a Ash 5.51±0.03a 5.45±0.01a 5.18±0.04b 5.28±0.06b 5.13±0.05b protein 13.5±0.34b 13.7±0.46b 13.4+0.35b 13.3±0.26b 15.1±0.12a crude fiber 5.4+0.1c 5.8±0.02c 9.3±0.08a 9.1±0.03a 8.1±0.12b crude fat 0.55±0.007b 0.52±0.008b 0.7±0.01a 0.67±0.002a 0.31±0.005c total sugar 55.8±1.3b 57.3±1.1b 68.6±1.4a 64.7±1.15a 46.7±1.55c reducing sugar 13.7±0.21b 14.9±0.31b 19.6±0.46a 18.3±0.5a 4.5±0.1c

3. Detection of mineral elements and some heavy metals

Determination method: use the inductively coupled plasma emission spectrometry in NY/T1653-2008 to determine the mineral elements and some heavy metal elements in the black fungus samples. Detection conditions: the carrier gas flow rate is 15L/min, the auxiliary gas flow rate is 0.2L/min, the pump flow rate is 1.5mL/min, the observation mode is axial, and the delay time is 15 seconds.

The test results are shown in Table 8. The content of Ca and Mg in black fungus HDXZ-1 (Auricularia auricular HDXZ-1) is the highest, and the analysis shows that the content of Ca and Mg in black fungus HDXZ-1 (Auricularia auricular HDXZ-1) is not only high Due to the local environmental factors in Tibet, black fungus HDXZ-1 (Auricularia auricular HDXZ-1) has a stronger ability to absorb Ca and Mg than the other four samples.

The content of P in black fungus HDXZ-1 (Auricularia auricular HDXZ-1) was the highest, while the transplanted varieties No. 3 and No. 4 samples were lower, indicating that although the P content in the culture environment was low, black fungus HDXZ-1 (Auricularia auricular HDXZ- 1) It has good absorption and enrichment ability for P.

The test shows that the absorption ability of black fungus HDXZ-1 (Auricularia auricular HDXZ-1) to K element is obviously better than that of No. 3 and No. 4 samples.

The test shows that the Zn content in the environment in Tibet is relatively high, and black fungus HDXZ-1 (Auricularia auricular HDXZ-1) has the function and effect of balancing Zn content.

The content of Na in black fungus HDXZ-1 (Auricularia auricular HDXZ-1) is the lowest.

The content of heavy metals Cr and Pb in black fungus HDXZ-1 (Auricularia auricular HDXZ-1) is the lowest, although the content of heavy metals Cr and Pb in the Tibetan plateau environment is high, black fungus HDXZ-1 (Auricularia auricularHDXZ-1) can effectively Avoid the absorption of these two harmful heavy metals.

The intake of Na and K must maintain a balance to be healthy for the human body; Ca, Zn, and P are all essential elements for the structure and function of bone cells, and promote bone formation; Mg is an activator of various enzymes in physiological and biochemical reactions, and Mg It also has the function of regulating muscle excitability; while Cr and Pb are heavy metals harmful to human body. Therefore, it can be seen that Auricularia auricular HDXZ-1 (Auricularia auricular HDXZ-1) is a kind of Auricularia auricularia strain suitable for planting in the Tibetan plateau area with reasonable mineral element content.

Table 8 Contents of mineral elements and heavy metal elements in fungus samples (μg/g)

name 1 2 3 4 5 Ca 3186±56 3245±65 4157±48 4229±36 4407±37 Zn 1.12±0.22 0.05±0.01 23.1±0.74 24.6±0.43 5.12±0.21 P 3187±15 3223±27 3019±27 3089±26 3658±23 K 12612±85 12481±101 10841±63 11089±74 12151±67 Na 1993±23 1960±17 1307±27 1249±12 889±15 Mg 1021±12 1323±28 1697±37 1773±35 2113±28 Cr 36.8±1.23 35.8±1.01 51.2±1.56 52.8±1.03 30.4±1.4 Pb 1.02±0.2 0.84±0.1 3.49±0.26 3.89±0.5 0.45±0.1 As 0.47±0.1 0.37±0.05 - - -

4. Vitamin content detection

Black fungus HDXZ-1 (Auricularia auricular HDXZ-1) has the highest VB ₂ content.

Claims (1)

1. the former black fungus strain of a plant height, it is woodear HDXZ-1(Auricularia auricular HDXZ-1), being deposited in Chinese Typical Representative culture collection center, deposit number is CCTCC NO:M2013032.