CN113455288A - Breeding method of disease-resistant ganoderma lucidum spore powder strain - Google Patents
Breeding method of disease-resistant ganoderma lucidum spore powder strain Download PDFInfo
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- CN113455288A CN113455288A CN202110868265.3A CN202110868265A CN113455288A CN 113455288 A CN113455288 A CN 113455288A CN 202110868265 A CN202110868265 A CN 202110868265A CN 113455288 A CN113455288 A CN 113455288A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/40—Cultivation of spawn
Abstract
The invention is suitable for the field of strain breeding methods, and provides a disease-resistant ganoderma lucidum spore powder strain breeding method, which comprises the following steps: selecting a strain: artificially domesticating wild ganoderma lucidum, and then selecting a disease-resistant strain according to the growth speed of hypha, the colony morphology, the high-temperature resistance and the inoculation survival rate; research on the disease resistance of the disease-resistant strains: in a ganoderma disease area, using a cut-log culture medium to research the disease resistance of strains in three stages of a ganoderma germination period, a pileus period and a spore production period of ganoderma, and breeding disease-resistant strains; research on application effects: carrying out a productivity test on the bred disease-resistant strains, and checking the practicability of the disease-resistant effect of the strains; the strain spore powder obtained by the breeding method of the invention has high yield and good capability of propagating the spore powder; the strain bred by the method has high survival rate in the ganoderma germination stage, pileus stage and spore production stage, and has good disease resistance.
Description
Technical Field
The invention belongs to the field of strain breeding methods, and particularly relates to a breeding method of disease-resistant ganoderma lucidum spore powder strains.
Background
Ganoderma (Ganoderma lucidum) Basidiomycetes order Polyporales family Polyporaceae family Ganoderma genus fungi. Is an effective and nontoxic top grade product of the traditional Chinese medicine in China, namely an ancient medical book Shen nong's herbal Jing in the period of eastern Han. The spore powder is Ganoderma lucidum germ cell, and has Ganoderma lucidum genetic active substance. In recent 60 years, the medicinal value of the spore powder is concerned by the market, the cultivation area is larger and larger, and the disease damages the ganoderma lucidum cultivation more and more. Research shows that in China, 110 edible fungus diseases comprise four major types of fungi, including fungi, viruses, bacteria and slime bacteria, main diseases of lucid ganoderma comprise penicillium, green mould, and a woody and simple type of fungi, which occur in various stages of lucid ganoderma cultivation, the lucid ganoderma buds are infected in the growth period, fungus material nutrients are consumed, toxins are secreted, hypha development is inhibited, lucid ganoderma sporocarp, stipe and lower layer porous tissues of pileus can be infected by the fungi, the sporocarp is swallowed, the yield of spore powder is reduced, and the ganoderma is harvested absolutely.
The disease prevention and cure of ganoderma lucidum has the measures of breeding disease-resistant strains, regulating and controlling cultivation conditions, preventing and curing medicaments and the like, and because ganoderma lucidum is also fungi, the selection of medicaments is greatly limited, and medicaments effective to pathogenic bacteria also have the effects of phytotoxicity and inhibition on the ganoderma lucidum fungi. The food is safe and does not allow the use of harmful medicaments for the ganoderma lucidum products. The most effective method for preventing and treating the ganoderma lucidum diseases is to select and apply disease-resistant strains. However, the existing ganoderma lucidum strains have no disease-resistant excellent strains and no breeding method of disease-resistant ganoderma lucidum spore powder strains. The breeding of disease-resistant strains is a technical problem which is urgently required to be solved for the production and development of ganoderma lucidum spore powder.
Disclosure of Invention
The embodiment of the invention aims to provide a breeding method of disease-resistant ganoderma lucidum spore powder strains, and aims to solve the problems in the background technology.
In order to achieve the above object, the embodiment of the present invention provides a method for breeding disease-resistant ganoderma lucidum spore powder strains, comprising the following steps:
selecting a strain: artificially domesticating wild ganoderma lucidum, and then selecting a disease-resistant strain according to the growth speed of hypha, the colony morphology, the high-temperature resistance and the inoculation survival rate;
research on the disease resistance of the disease-resistant strains: in a ganoderma disease area, using a cut-log culture medium to research the disease resistance of strains in three stages of a ganoderma germination period, a pileus period and a spore production period of ganoderma, and breeding disease-resistant strains;
research on application effects: and (4) carrying out a productivity test on the bred disease-resistant strains, and checking the practicability of the disease-resistant effect of the strains.
Preferably, the operation steps of the artificial domestication of the wild ganoderma lucidum are as follows: collecting wild Ganoderma, tissue separating, and culturing strain.
Preferably, the conditions for collecting the wild ganoderma lucidum are as follows: the average daily temperature of the collected land is more than or equal to 10 ℃, the effective annual accumulated temperature is more than or equal to 5700 ℃, the annual rainfall capacity is more than 1650mm, the forest coverage rate is more than 70 percent in subtropical mountain areas, and the red ganoderma grows on rotten wood stumps; ganoderma pileus is 10-15cm, white growth circle is 3cm, thickness of the pileus is more than 1cm, bottom is beige, and developed strong Ganoderma is obtained.
Preferably, the tissue separation: cutting the basal part of Ganoderma stipe, soaking in 75% of the essence, sterilizing cotton, removing peel and shell of fruiting body with surgical knife, cutting into granules with soybean size, sterilizing with flame, and transferring to pDA tube for culture.
Preferably, the cultured strain is: six strains A301-A306 are obtained by tissue isolation; transferring slant mycelia, culturing in 27 deg.C culture room for 8 days, and testing mycelia shape.
Preferably, the top of the ganoderma lucidum sprout period is white and bright, the ganoderma lucidum grows strongly, the survival is evaluated, the top is grey black, and the ganoderma lucidum sprout atrophy is evaluated to die;
in the pileus period, the white growth ring covered with the cap is bright, the pileus grows strongly, survival is evaluated, the growing ring covered with the cap is gray and black, the pileus shrinks, and death is evaluated;
in the spore production period, when the temperature in the greenhouse is 25-29 ℃, a large amount of spores are sprayed to the fungus tube holes below the pileus, the bottom of the pileus is yellow, the survival is evaluated, the fungus tube holes below the pileus are not sprayed with spores, and the bottom of the pileus is grey black, and the death is evaluated.
Preferably, in the process of researching the disease resistance of the disease-resistant strains, the ganoderma is cultivated in a ganoderma disease area for more than three years, organic substances and soil in the environment exist, a large amount of penicillium, streptomyces viridis, trichoderma and other germs exist in the air, and a log culture medium is used.
In summary, due to the adoption of the technical scheme, the method has the following beneficial effects:
the embodiment of the invention provides a breeding method of disease-resistant ganoderma lucidum spore powder strains, and the strains obtained by the breeding method have high spore powder yield and good spore powder propagation capacity; the strain bred by the method has high survival rate in the ganoderma germination stage, pileus stage and spore production stage, and has good disease resistance.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the embodiments of the present invention are described in further detail below with reference to specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the embodiments of the invention and are not limiting of the embodiments of the invention.
Example 1
Collecting wild ganoderma lucidum: the collected land has an average daily temperature of 15 ℃, an effective annual accumulated temperature of 5800 ℃, an annual rainfall of 1650mm or more, a forest coverage rate of more than 70% in subtropical mountain areas, and ganoderma lucidum growing on rotten wood stumps. Ganoderma pileus 15cm, white growth circle 3cm, cover thickness more than 1cm, bottom beige, and developed strong Ganoderma.
Tissue isolation: cutting the basal part of Ganoderma stipe, soaking in 75% of the essence, sterilizing cotton, removing peel and shell of fruiting body with surgical knife, cutting into granules with soybean size, sterilizing with flame, and transferring to pDA tube for culture.
Strain test and tissue separation to obtain six strains A301-A306. Transferring slant mycelia, placing in 27 deg.C culture room, and culturing for 8 days to test mycelia shape. The results are shown in Table 1.
TABLE 1
| Numbering | Form of the composition | Colour(s) | Density of | Growth speed arm spread (cm/d) |
| A301 | Is tiled and attached to | White colour | ++++ | 1.98 |
| A302 | Tiling | White colour | +++ | 1.65 |
| A303 | Fluffy shape | White colour | +++ | 1.38 |
| A304 | Tiling | White colour | ++ | 1.92 |
| A305 | Tiling | Grey colour | ++ | 1.30 |
| A306 | Is tiled and attached to | White colour | ++++ | 1.93 |
The test shows that: ganoderma acclimatization A301 mycelium has good shape, color, density, and development speed, and mycelium growth speed of 1.98 (cm/d).
The spore-forming ability of the strain was investigated by comparing A301 with the strain B201 used in production. The results are shown in Table 2.
TABLE 2
| Strain numbering | Number of bacterial material segments | Weight kg of fungus material | Lucid ganoderma 'duo' shape | Collected spore powder kg | 100kg of cut-log spore powder with the yield of kg | Comparison of |
| B201 | 30 | 240 | Circular shape with cover diameter of 10-13cm and beige bottom | 44.4 | 1.85 | 100 |
| A301 | 30 | 240 | The diameter of the cover is 13-15cm, and the sector bottom color is golden yellow | 52.8 | 2.20 | 118.9 |
Tests show that; the yield of the spore powder of 100kg of the log is 18.9 percent higher than that of a contrast strain, and the spore powder of A301 has good spore reproduction capability.
And carrying out disease resistance test on the bred spore powder strains in the disease area.
Test site: in a ganoderma disease area, ganoderma is cultivated for more than three years, organic substances and soil in the environment allow the ganoderma to grow, and a large amount of penicillium, streptomyces viridis, trichoderma and other germs exist in the air; cut-log culture medium was used.
The test strains were: a301 domesticating wild ganoderma; control strain B201 was a locally applied producer strain.
The test method comprises the following steps:
according to the technical specification of the cut-log ganoderma lucidum production, the procedures of tree seed selection, fungus section production, bagging, sterilization and the like are completed, the weight of the fungus section is 8Kg, and test strains A301 and B201 are inoculated to the cut-log for fungus growing.
A301 and B201 are respectively selected to have good spawn running and 50 sections without mixed bacteria pollution, and are subjected to a soil-buried culture test.
Building a sesame shed with the height of 2m, covering a nylon membrane, and covering a sunshade net; measures such as water spraying, ventilation and the like are taken well, the temperature and the humidity in the greenhouse are adjusted, the temperature is controlled to be 28 ℃, and the relative humidity of air is 85%.
Pruning when Ganoderma lucidum grows Ganoderma lucidum sprout, reserving one Ganoderma lucidum sprout in one section of the fungus material, and cultivating one Ganoderma lucidum. The disease resistance of the strain is tested in three stages of ganoderma, pileus and spore production.
The disease resistance of the strain is determined, the top of the ganoderma bud stage is white and bright, the strain grows healthily and strongly, and the survival is evaluated; the top end is gray black, and the ganoderma sprouts are atrophied and evaluated to die. In the pileus period, the white growing ring at the cover edge is bright, the pileus grows strongly, and the survival is evaluated; the growth circle of the cap edge is gray black, the pileus is atrophied, and the death is evaluated. In the sporulation period, when the temperature in the greenhouse is 28 ℃, a large amount of spores are sprayed in the fungus tube holes below the pileus, the bottom of the pileus is yellow, and survival is evaluated; the pore of the fungus tube under the pileus is not sprayed with spores, and the bottom of the pileus is evaluated to be dead in gray and black.
Calculating the yield of the spore powder: the lucid ganoderma sporocarp grows mature, white edges disappear, spore powder is cultivated by bundling a bag and a sleeve, the sleeve is harvested for 90 days, the yield of the spore powder is calculated according to the ratio of the dry weight of the spore powder to a cut-log culture medium, and the wet weight of the spore powder is dried by 85 percent.
Test results
The disease resistance of the strains is shown in table 3.
TABLE 3
The test shows that: the survival rate of the A301 is 55 percent higher than that of the contrast ganoderma B201 in the bud period through two-year cultivation experiments in the disease area. The cap period is 73.2 percent higher, and the sporulation period is 82.6 percent higher. A301 has good disease resistance.
The spore powder yields are shown in table 4.
TABLE 4
The test shows that: the yield of spore powder of the A301 strain through cut-log cultivation for two years is 33.2 percent higher than that of the control strain B201, and the disease resistance is good.
And (3) application effect inspection:
the disease-resistant strain A301 and the control strain B201 are used for cultivating cut-log 16000KG respectively, and the spore powder yield in two years is checked to calculate the cultivation benefit. The results are shown in Table 5
TABLE 5
The application effect shows that: 16000KG cut wood cultivation, disease-resistant strain A301 harvest spore powder 690KG two years, higher than the contrast strain 141.3%, the sales income 69000 yuan, higher than the contrast strain 164.37%, the income increase 20666 yuan, higher than the contrast strain 153.57%, has good economic and social benefits, the method has good practicability.
In summary, the following steps: the embodiment of the invention provides a breeding method of disease-resistant ganoderma lucidum spore powder strains, and the strain spore powder obtained by the breeding method is high in yield and has good spore powder breeding capability; the strain bred by the method has high survival rate in the ganoderma germination stage, pileus stage and spore production stage, and has good disease resistance.
It will be evident to those skilled in the art that the embodiments of the present invention are not limited to the details of the foregoing illustrative embodiments, and that the embodiments of the present invention are capable of being embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the embodiments being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (7)
1. A breeding method of disease-resistant ganoderma lucidum spore powder strains is characterized by comprising the following steps:
selecting a strain: artificially domesticating wild ganoderma lucidum, and then selecting a disease-resistant strain according to the growth speed of hypha, the colony morphology, the high-temperature resistance and the inoculation survival rate;
research on the disease resistance of the disease-resistant strains: in a ganoderma disease area, using a cut-log culture medium to research the disease resistance of strains in three stages of a ganoderma germination period, a pileus period and a spore production period of ganoderma, and breeding disease-resistant strains;
research on application effects: and (4) carrying out a productivity test on the bred disease-resistant strains, and checking the practicability of the disease-resistant effect of the strains.
2. The method for breeding disease-resistant ganoderma lucidum spore powder strains according to claim 1, wherein the operation steps of artificially domesticating wild ganoderma lucidum comprise: collecting wild Ganoderma, tissue separating, and culturing strain.
3. The method for breeding disease-resistant ganoderma lucidum spore powder strains according to claim 2, wherein the conditions for collecting the wild ganoderma lucidum are as follows: the average daily temperature of the collected land is more than or equal to 10 ℃, the effective annual accumulated temperature is more than or equal to 5700 ℃, the annual rainfall capacity is more than 1650mm, the forest coverage rate is more than 70 percent in subtropical mountain areas, and the red ganoderma grows on rotten wood stumps; ganoderma pileus is 10-15cm, white growth circle is 3cm, thickness of the pileus is more than 1cm, bottom is beige, and developed strong Ganoderma is obtained.
4. The method for breeding disease-resistant ganoderma lucidum spore powder strains according to claim 3, wherein the tissue isolation: cutting the base of Ganoderma stipe, spraying essence, soaking, sterilizing cotton, cleaning, peeling with surgical knife, cutting into soybean granules, sterilizing with flame, and transferring to pDA tube for culture.
5. The method for breeding disease-resistant ganoderma lucidum spore powder strains according to claim 4, wherein the cultured strains are: six strains A301-A306 are obtained by tissue isolation; transferring slant mycelia, culturing in 27 deg.C culture room for 8 days, and testing mycelia shape.
6. The method for breeding disease-resistant ganoderma lucidum spore powder strains according to claim 5, wherein the top of the ganoderma lucidum sprout period is white and bright, the top of the ganoderma lucidum sprout period grows strongly, survival is assessed, the top is gray and black, and ganoderma lucidum sprout atrophy is assessed to die;
in the pileus period, the white growth ring covered with the cap is bright, the pileus grows strongly, survival is evaluated, the growing ring covered with the cap is gray and black, the pileus shrinks, and death is evaluated;
in the spore production period, when the temperature in the greenhouse is 25-29 ℃, a large amount of spores are sprayed to the fungus tube holes below the pileus, the bottom of the pileus is yellow, the survival is evaluated, the fungus tube holes below the pileus are not sprayed with spores, and the bottom of the pileus is grey black, and the death is evaluated.
7. The method for breeding disease-resistant ganoderma lucidum spore powder strains according to claim 6, wherein the test site for studying the disease resistance of the disease-resistant strains is that ganoderma lucidum is cultivated in a ganoderma lucidum disease area for more than three years, organic substances and soil in the environment exist, a large amount of penicillium, streptomyces viridis, trichoderma and other germs exist in the air, and a log culture medium is used.
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