CN112080436B

CN112080436B - Large fungus strain with high cadmium resistance and application thereof

Info

Publication number: CN112080436B
Application number: CN202011052123.1A
Authority: CN
Inventors: 李小方; 陈苗苗; 郑鑫
Original assignee: Institute of Genetics and Developmental Biology of CAS
Current assignee: Institute of Genetics and Developmental Biology of CAS
Priority date: 2020-09-29
Filing date: 2020-09-29
Publication date: 2022-01-04
Anticipated expiration: 2040-09-29
Also published as: CN112080436A

Abstract

The invention provides a large fungus strain with high cadmium resistance and application thereof, wherein the strain is named as Pleurotus ostreatus (Pleurotus ostreatus) pleur-jn 21-2F1, is preserved in China general microbiological culture Collection center on 08-11 months in 2020, and has a preservation number of CGMCC No. 20239. The invention also discloses application of the strain in repairing cadmium-containing polluted soil or water. The large-scale fungus strain pleur-jn 21-2F1 provided by the invention is used for repairing cadmium-containing soil or water, the operation process is simple, the cost is low, the strain has a good cadmium enrichment effect, and the soil or water repairing efficiency can be improved.

Description

Large fungus strain with high cadmium resistance and application thereof

Technical Field

The invention belongs to the technical field of heavy metal pollution remediation, and particularly relates to a high-cadmium-resistance large-scale fungus strain and application thereof.

Background

With the rapid development of modern industrialization, soil and water in China are polluted by heavy metals, wherein cadmium is one of the most important elements in heavy metal pollution. Cadmium is a highly toxic pollutant, and in recent years, many cadmium pollution events have occurred in China, and the health of human beings is seriously influenced.

In recent years, the biological technology is more and more concerned about repairing cadmium-containing soil or water, and the method is safer, more environment-friendly and cheaper than chemical and physical repairing methods. At present, the biological repair of cadmium-containing soil or water mainly depends on plant varieties, the cadmium content in the soil is reduced by screening and culturing the plant varieties with high cadmium resistance and absorbing cadmium element in the growth process of the plant varieties, and thus the repair of the cadmium-containing soil or water is realized. Meanwhile, the plant varieties with high-efficiency heavy metal enrichment characteristic and high growth speed are few, the screening and culturing process generally consumes a long time, and the cost is high. In the existing research, the macrofungi are used for repairing cadmium-containing soil or water body, so that the repairing time is greatly shortened, but the repairing efficiency needs to be further improved due to the limitation of the tolerance and enrichment capacity of the strains. Therefore, a novel macro fungus with strong resistance and strong adsorption capacity is urgently needed to be used for repairing cadmium-containing soil or water so as to reduce the cadmium content in the soil or water.

Disclosure of Invention

Aiming at the problems in the prior art, the invention provides the high-cadmium-resistance large fungus strain and the application thereof.

In order to achieve the purpose, the technical scheme adopted by the invention for solving the technical problems is as follows:

a large-scale fungus strain with high cadmium resistance is obtained by a Cd concentration gradient directed mutagenesis method, is named as Pleurotus ostreatus (Pleurotus ostreatus) pleur-jn 21-2F1, is preserved in China general microbiological culture Collection center (CGMCC) at 11/08 in 2020, and has a preservation number of CGMCC No. 20239.

The invention also provides application of the large fungus strain pleur-jn 21-2F1 in repairing cadmium-containing polluted soil or water.

When the strain is used for cadmium-containing soil, a culture medium (a mushroom stick) with hyphae full of large-scale fungus strains pleur-jn 21-2F1 can be buried in the soil for fruiting management, heavy metals are enriched into mushroom bodies and the culture medium (the mushroom stick) along with the growth of thalli, and finally the heavy metals are removed by harvesting the mushroom stick and the thalli, so that the effect of repairing can be achieved; when the mycelium is used for water body restoration, the mycelium obtained by culture can be directly added into the water body, and the restoration effect can be achieved. The method comprises the following specific steps:

when the cadmium-containing soil is repaired, a large-scale fungus strain pleur-jn 21-2F1 is inoculated in a solid PDA culture medium for activation, activated mycelium is inoculated in a culture medium for culture to obtain a fungus stick, the fungus stick is subjected to primordium induction, and after the primordium appears, the fungus stick is moved into the cadmium-containing soil for fruiting management.

When the water body is restored, the large-scale fungus strain pleur-jn 21-2F1 is inoculated in a solid PDA culture medium for activation, the activated fungus block is inoculated in a liquid PDA culture medium again to obtain a mycelium pellet, and the mycelium pellet is directly added into the cadmium-containing water body to play a role in restoration.

The high-cadmium-resistance large fungus strain and the application thereof provided by the invention have the following beneficial effects:

the large-scale fungus strain pleur-jn 21-2F1 provided by the invention is used for repairing cadmium-containing soil or water, the operation process is simple, the cost is low, the strain has a good cadmium enrichment effect, and the soil or water repairing efficiency can be improved.

Drawings

FIG. 1 is a concentric disc Cd gradient medium.

FIG. 2 is a graph of comparative culture results of resistance mutation primary selection strains and original strains under Cd stress.

FIG. 3 is a diagram showing the result of ISSR analysis of a mutant strain.

FIG. 4 is a graph showing the growth of the original strain and the high Cd-resistant mutant strain 2F 1.

FIG. 5 shows the growth of the original strain and the highly Cd-resistant mutant strain 2F1 at different Cd concentrations.

Detailed Description

Example 1

First, test method

1. Test strains

The original strain used in the invention is a high-cadmium resistant oyster mushroom strain obtained by early screening in a laboratory, and is registered and preserved in China general microbiological culture Collection center (CGMCC) at 7 and 6 days in 2018, with the preservation number of CGMCC NO. 16084.

2. Strain activation

Transferring the test strain to solid PDA culture medium, activating and culturing at 28 deg.C, and continuously transferring for 3 times.

Wherein 1L of solid PDA culture medium contains potato soaking powder 3g, glucose 20g, and agar powder 15g, and the pH is adjusted to 6.0 + -0.2.

3. Preparation of heavy metal mother liquor

Accurately weigh 10.16g CdCl₂·2H₂Dissolving O in 10mL deionized water (reagent is analytically pure), filtering with 0.22 μm filter membrane (to remove microorganisms in water) to obtain heavy metal mother liquor with concentration of 500g/L, and storing at 4 deg.C for use.

4. Directed mutagenesis and primary screening of Cd-resistant mutant strains

The directional mutagenesis adopts a concentric disc Cd concentration gradient culture medium (see figure 1), and the preparation method comprises the following steps:

(1) preparing a sterilized petri dish (diameter 19 cm); preparing sterilizable circular molds with different sizes for the isolation between the molds and different culture mediums when manufacturing concentric circular culture mediums, and sterilizing at high temperature and high pressure.

(2) Preparing a PDA culture medium, carrying out autoclaving at 121 ℃ for 20min, adding heavy metal mother liquor with different volumes when the culture medium is cooled to about 65 ℃, and preparing the culture medium with Cd concentrations of 0, 20, 80, 160 and 240 mg/kg.

(3) Preparation of concentric disc culture medium: pouring a thin layer of Cd-free solid culture medium at the bottom of a sterilized culture dish, inserting sterilized ring molds with different diameters into the thin layer culture medium with the center of the culture dish as the center of a circle after the culture medium is solidified, and pouring culture media with Cd concentrations of 0, 20, 80, 160 and 240mg/kg from the center circle to the outer ring in sequence to prepare the concentric circle Cd gradient mutagenesis culture medium. The thin-layer solid culture medium at the bottom can prevent the pollution among the culture media caused by the leakage of the unclosed bottom when different culture media are poured into the culture dish. Meanwhile, in order to avoid the situation that the bottom sealing culture gene is dissolved due to long-time large-area heating, a mode of pouring another culture medium after one culture medium is solidified is adopted. And taking out the mold ring after the culture medium is completely solidified.

(4) Activated strain CGMCC NO.16084 (earlier stage experiment proves that the strain can not grow under 160mg/kg Cd) is inoculated to the center position of concentric circle culture medium by 0.5cm inoculation block, and is cultured in dark at 28 ℃ to extend hyphae to the edge. Mutagenesis was terminated when hyphae extended to the most marginal maximum Cd concentration.

(5) When hypha or colony is amplified to a culture medium containing a mutagen with the maximum tolerance concentration higher than that of an original strain, a point is projected along the center of a circular ring to the outer edge for radiation scribing, a bacterium block with the diameter of 0.5cm is inoculated to a PDA culture medium with Cd 160mg/kg along each concentration line, a resistance-enhanced strain is preliminarily screened, the preliminarily selected mutant strain is inoculated to the PDA culture medium with the Cd concentration of 120mg/kg, the original strain is simultaneously inoculated to the same culture dish, and the strain is subjected to dark culture at 28 ℃ for 20 days. A positively mutated colony with increased resistance is initially judged if it is able to grow and is significantly better than the original strain.

5. Molecular identification of cadmium-resistant positive mutant strain

ISSR (Simple Repeat Sequence Repeat) molecules identify mutants that are used to determine structural variations in a gene. The specific operation is as follows.

Continuously transferring all positive mutants twice on a PDA plate with Cd of 160mg/kg, after the characters are stable, extracting hyphal DNA (plant genome DNA extraction kit DP305, TIANGEN) of each positive strain and an original strain (CGMCC NO.16084), adopting different ISSR primer sequences, taking the DNA of the original strain and the mutant strain as templates, and carrying out PCR amplification to obtain the mutant strain with a gene structure having variation. The ISSR primer sequences are shown in the following table.

TABLE 1 Strain ISSR primer sequences

6. Evaluation of high cadmium resistance Positive mutant strains

6.1 determination of growth Curve of Cd-resistant mutant

After the determined high Cd resistance mutant strain is activated and cultured in a common PDA plate, a 0.5cm inoculation block is inoculated in the center of a common PDA culture medium plate (inner diameter is 8.5cm), the original strain is used as a control, each strain is arranged in three times, and the strain is cultured in a dark place at 28 ℃. Hypha growth was observed daily and colony diameters were recorded until the plates were covered with hyphae. And drawing a strain growth curve and determining the growth speed.

6.2 maximum tolerated concentration evaluation of Cd

The maximum tolerated concentration refers to the critical concentration of the mycelium which is tolerated by certain heavy metals, i.e.at the maximum tolerated concentration there is growth of the mycelium, slightly above which growth is completely inhibited.

Preparing a PDA culture medium, carrying out autoclaving at 121 ℃ for 20min, adding heavy metal mother liquor with different volumes when the culture medium is cooled to about 65 ℃, and preparing 20mL PDA solid plates with Cd concentrations of 0, 10, 160, 240, 280, 320, 360, 400, 440 and 480 mg/kg.

The activated original strain (CGMCC NO.16084) and the positive mutant strain are inoculated into a culture dish (inner diameter 8.5cm) containing Cd concentration by taking 0.5cm inoculation blocks at the hypha growth tip respectively, and each strain is provided with 3 times of repetition at each concentration. The cells were incubated at 28 ℃ for 24 days in the dark and the colony diameter was recorded for each strain at each concentration.

Second, test results

1. Preliminary determination of Cd-resistant mutants

6 mutants with enhanced resistance are obtained in total by primary resistance screening in a PDA culture medium with Cd concentration of 160mg/kg, namely 2F1-3b-2, 2F1-3b-9, 2F1-3d-1, 2F1-3d-2, 2F1-3d-3 and 2F1-3 d-7. 6 resistant mutant strains are respectively cultured for 20 days in a PDA culture medium with the Cd content of 120mg/kg with the original strain in a confronting manner (see figure 2, the left part of each culture dish is a mutant strain, the right part of each culture dish is an original strain, the three culture media at the upper part are respectively 2F1-3b-2, 2F1-3b-9 and 2F1-3d-1 from left to right, and the three culture media at the lower part are respectively 2F1-3d-2, 2F1-3d-3 and 2F1-3d-7 from left to right), the colony diameter of the mutant strain is obviously larger than that of the original strain, which indicates that the Cd resistance of the mutant strain is obviously enhanced.

2. Obtaining of high Cd resistant mutant strain 2F1

Through ISSR molecular identification, the amplification results of ISSR primers P7 and P24 (see the amplification results of primers P7 and P24 in figures 3 and A, B respectively) show that electrophoresis bands of 6 mutant strains are obviously deleted compared with the original strain, which indicates that the 6 strains have gene structure variation and the Cd resistance of the strain is enhanced due to the gene structure variation. However, there was no significant difference in electropherograms between the 6 mutant strains, that is, the mutation sites of the 6 mutants may be less different or the same. Therefore, we considered the 6 mutants as one strain, the strain is Pleurotus ostreatus (Pleurotus ostreatus) pleur-jn 21-2F1 (hereinafter referred to as 2F1), and the Pleurotus ostreatus (Pleurotus ostreatus) Pleurotus ostreatus is preserved in China general microbiological culture Collection center (CGMCC No. 20239) at 11/08 in 2020.

3. Growth rate of high Cd resistant mutant strain 2F1 is reduced

The growth rate of the obtained high Cd resistance strain 2F1 is tested, and a growth curve of the strain is constructed. The results showed that the 2F1 mutant strain grew significantly slower than the original strain (see FIG. 4).

4. The resistance of the mutant strain 2F1 Cd is obviously enhanced

The experimental results show (see FIG. 5) that the original strain stopped growing at a Cd concentration of 160mg/kg (inoculum size of 0.5cm, i.e.colony diameter of 0.5cm, indicating no growth of the pellet), whereas the mutant strain 2F1 stopped growing at a Cd concentration of 480mg/kg, which increased the Cd resistance by a factor of 3 compared to the original strain. The mutant strain 2F1 had a maximum Cd tolerance concentration of 440 mg/kg.

5. The resistance of the mutant strain 2F1 Cd is higher than the average level of other large fungi

The Cd resistance capability (maximum tolerant concentration 440mg/kg) of the mutant strain 2F1 is far higher than the Cd resistance level of other large fungi, and is only lower than a strain Pleurotus Sp (maximum tolerant concentration of Cd 700mg/kg) separated beside a municipal refuse pile, possibly as a result of long-term evolution of the strain in a polluted environment.

TABLE 2 Macro-fungal mycelium Cd tolerance

Test example 1 remediation of cadmium soil

First, test method

1. Soil treatment

1.1 soil preparation

Air drying the soil, grinding, sieving with a 2mm sieve, and removing impurities from the soil.

1.2 soil pH determination

5g of dry soil is taken in a 50mL centrifuge tube, 25mL of ultrapure water is added, the mixture is inverted from top to bottom for several times and then is kept stand for 1h, and when the solution is clear, a pH meter is used for measuring the pH value in a supernatant.

1.3 maximum Water holding Capacity determination

50g of the prepared soil were taken in a funnel with filter paper, a hose was added under the funnel, and the hose was held by a clip. 50mL of water was poured into the soil, and after the water was equilibrated in the soil for 1 hour, the clamp was opened to allow the water to naturally flow out, and collected. And finishing water collection when the water does not drip out. Maximum water holding capacity of soil (initial water addition volume-final water collection volume)/initial soil mass.

1.4 soil Cd treatment

Accurately weighing a certain amount of soil, adding a Cd solution, and fully mixing to ensure that the Cd content of the soil is 0, 60, 160, 240, 320, 360, 400, 420 and 440mg/kg respectively. And (4) after the soil is balanced for 30 days, measuring the pH of the soil with each Cd concentration for later use.

2. Strain preparation

2.1 preparation of PDA Medium (1L)

Taking 3g of potato extract powder, 20g of glucose and 15g of agar powder, adding 900mL of distilled water, mixing uniformly, adjusting the pH to 6.0 +/-0.2, adding distilled water to a constant volume of 1000mL, and sterilizing at 121 ℃ for 20min under high pressure.

2.2 Strain activation

The strain 2F1 which is preserved at low temperature is inoculated to the center of a PDA culture medium and is cultured in the dark at 28 ℃.

2.3 obtaining of mycelium

Inoculating 0.5cm diameter inoculating block in PDA culture medium, and culturing at 28 deg.C in dark for 7 days to obtain mycelium material.

3. Preparation of fungus stick

3.1 formula and preparation of culture medium

Uniformly mixing 78% of cottonseed hull, 20% of bran, 1% of gypsum and 1% of cane sugar to obtain a culture medium, adding water which is 1.6 times of the culture medium in weight, fully and uniformly mixing, and naturally standing for 30min to enable water to fully permeate into the culture medium. Drying a small amount of mixed matrix, measuring Cd content, packaging the rest of the mixed matrix into 130 × 240 × 0.045mm polypropylene bags, compacting, punching the center of the matrix, fastening the bag mouth, wetting the matrix in each bag to a weight of 350 + -5 g, and autoclaving at 121 deg.C for 100 min.

3.2 inoculation and culture of the fungus sticks

And (3) after the substrate is cooled, inoculating the cultured inoculation block containing the mutant strain 2F1 mycelium into the substrate, tying the fungus bag, and culturing at the constant temperature of 28 ℃ in a dark place for 28 days to obtain the culture fungus stick.

4. Potted plant

4.1 transferring the fungus sticks full of fungus into a mushroom house, wherein the temperature of the mushroom house is 18 ℃, the humidity is 80-85%, the mushroom house is adapted for 2 days, and the fungus sticks are opened at the tying position for primordium induction. The temperature of the primordium induction is 18 ℃, the humidity is 80-85%, the primordium is ventilated for 1 hour in the morning and at night every day, the temperature is 5-8 ℃, and the humidity is about 50%. And (5) allowing the plant to appear with primordium, and performing potting treatment.

4.2 potting treatment

Selecting fungus sticks with the same primordium size, cutting off 2/3 fungus bags, transferring into a flowerpot with the inner diameter of 210 x 160mm, placing in the center of the flowerpot, filling the rest part of the flowerpot with Cd to treat well-balanced soil, and adopting a semi-underground cultivation mode for the fungus sticks, wherein the part without the fungus bags is exposed outside. 1080g of balanced dry soil is added into each pot, and water is added to reach 65 percent of the maximum water holding capacity. Inserting a soil moisture collector into the soil 1cm away from the fungus rods, wherein 1 soil moisture collector is arranged in each pot, and taking 5mL of soil aqueous solution every 7 days from the first day when the fungus rods enter the soil to the third crop of sporocarp harvest, wherein the concentration of each Cd is set to be 3 times.

4.3 potted plant culture

And (4) after the fungus sticks are buried, performing fruiting management and keeping the fruiting management. Keeping the temperature of the mushroom growing room at 18 ℃ and the humidity of 85-95%, and ventilating every morning and evening for 0.5 h. And (4) after harvesting the fruiting bodies of each tide, numbering the fruiting bodies, placing the numbering on the fruiting bodies in an oven, drying, weighing, and simultaneously performing primordium induction and fruiting management on the next tide of mushrooms. After harvesting the three-tide sporocarp, taking out the fungus stick from the soil and drying the fungus stick for measuring the content of Cd; and collecting and drying soil in each basin, and measuring pH.

5. Analysis of Cd content

And taking 0.3g of each dried fruiting body for processing each harvested stubble, the culture medium before inoculation and the culture medium after harvesting, transferring the fruiting bodies into a digestion tube, adding 8mL of 65% nitric acid, soaking overnight, placing the digestion tube on an electric heating plate for digestion, adding a funnel for digestion for 24 hours, then removing acid at high temperature, adding 5mL of 65% nitric acid, and continuously heating until the mixture is evaporated to dryness. Each sample was digested in 3 replicates with 5mL of ultrapure water. The analysis of the Cd content was measured with a Zeenit 700P atomic absorption spectrophotometer flame method.

6. Data analysis

Data analysis was performed using SPSS 16.0. Data comparison was performed using one-way anova with a test standard P < 0.05.

Second, test results

1. Maximum water holding capacity of soil and growth of fruiting body

The maximum water holding capacity of the soil was determined to be 47%.

After the fungus sticks enter the fruiting chamber, 8 days are needed for each tide of fruiting body primordium induction, and 8 days are needed for fruiting body growth. The first tide of fruit body is moved into the soil after the primordium is shown, and the third tide of fruit body is harvested with the fungus stick in the soil for 40 days.

2. The test soil was alkaline soil with a pH of 8.6. The strain 2F1 can reduce the pH value of soil by about 0.5 by culturing fruiting bodies. The pH value of the soil before and after Cd treatment is not obviously different, but the pH value of the soil after fruiting is obviously reduced, so that the growth process of the strain can reduce the pH value of the soil, increase the effective state Cd in the soil, and is beneficial to the absorption of the strain on the Cd in the soil, thereby playing a role in restoring alkaline soil.

3. The enrichment capacity of the fruiting bodies is increased along with the increase of the harvest tide number and is increased along with the increase of the concentration of Cd in the soil, and specific results are shown in a table 3.

TABLE 3 amount of cadmium enrichment of different hygromycin mushrooms at different soil cadmium treatment concentrations

Cd concentration (mg/kg)	First tide mushroom (mg)	Second tide mushroom (mg)	Third tide mushroom (mg)
				10	0.004	0.008	0.010
160	0.007	0.018	0.012
				240	0.009	0.021	0.013
280	0.012	0.028	0.014
				320	0.013	0.032	0.014
360	0.015	0.038	0.016
				400	0.016	0.040	0.017
420	0.017	0.042	0.016
				440	0.020	0.045	0.017
480	0.019	0.040	0.015

The cadmium-enriched amount of each tide of mushroom is the amount of cadmium accumulated in the fruiting body produced by a single mushroom stick in each tide of mushroom.

As can be seen from Table 3, the cadmium enrichment capacity of the strain 2F1 is increased along with the increase of the harvest tide number, but the enrichment capacity of the third tide of mushrooms is inferior to that of the second tide of mushrooms, and a large amount of nutrients in the culture materials are consumed during fruiting of the first second tide of mushrooms, so that the overall metabolic capacity of the strain is influenced, and the cadmium enrichment capacity of the strain is further influenced. The strain 2F1 has strong enrichment capacity on cadmium, so the enrichment capacity of the fruiting body on cadmium is increased along with the increase of the concentration of Cd in soil.

4. The enrichment capacity of the bacteria sticks is increased along with the increase of the content of Cd in the soil, the concentration of cadmium in the soil is 160mg/kg, and the concentration of enriched Cd in a single bacteria stick is 12mg/kg within 40 days; when the concentration of cadmium in the soil is 420mg/kg, the concentration of enriched Cd in a single bacterium rod is 30mg/kg within 40 days; when the concentration of cadmium in the soil is 440mg/kg, the concentration of enriched Cd in a single bacterium rod is 34mg/kg within 40 days; when the concentration of cadmium in the soil is 480mg/kg, the concentration of enriched Cd in a single bacterium rod is 32mg/kg within 40 days.

5. The concentration of Cd in the soil aqueous solution shows a trend of rising firstly and then falling along with the increase of the culture days, and reaches the maximum in 32 days, which shows that Cd in the soil aqueous solution is in a dissociation state, which shows that the growth activity of the fruiting body can dissociate the Cd in the chelation state in the soil to enrich Cd in the fungus stick and the fruiting body, and the concentration of Cd in the soil aqueous solution is reduced due to the factors of enrichment, absorption and the like of the fungus stick and the fruiting body after the Cd in the soil aqueous solution is dissociated to a certain degree.

6. The strain 2F1 of the invention can still keep a good growth state in the soil with cadmium content of 0, 60, 160, 240, 320, 360, 400, 420, 440 and 480mg/kg respectively, and does not influence the growth and development of fruit bodies.

Test example 2 adsorption of cadmium in liquid by Strain 2F1

First, test method

1. Inoculating the activated strain 2F1 into a liquid PDA culture medium, adding cadmium with different concentrations of 0, 30, 60, 90, 120, 140 and 160mg/L, placing the culture medium at 28 ℃ for dark culture for 7 days, setting 3 parallel concentrations, taking supernatant, measuring the residual cadmium concentration, and calculating the removal rate.

Second, test results

When the cadmium concentration is in a certain range, the removal rate of the strain 2F1 to cadmium is increased along with the increase of the cadmium concentration, which indicates that the strain 2F1 has good adsorption to cadmium, but when the cadmium concentration is 140mg/L, the removal rate of the strain 2F1 to cadmium is the highest, which indicates that the tolerance concentration of the strain 2F1 to cadmium in liquid is 140mg/L, and the specific results are shown in Table 4.

TABLE 4 removal rate of cadmium by strain 2F1 under liquid culture conditions

Concentration of

30mg/L

60mg/L

90mg/L

120mg/L

140mg/L

160mg/L

Removal rate

3.5％

15.9％

20.1％

27.8％

52.1％

——

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Claims

1. A large fungus strain with high cadmium resistance is named Pleurotus ostreatus (Pleurotus ostreatus) pleur-jn 21-2F1, is preserved in China general microbiological culture Collection center (CGMCC) at 11/08/2020 with the preservation number of CGMCC No. 20239.

2. The use of the high cadmium resistant macrofungal strain of claim 1, pleur-jn 21-2F1, for remediation of soil or water contaminated with cadmium.

3. The application of claim 2, wherein when repairing cadmium contaminated soil, the soil repairing effect can be achieved by burying a culture medium of a large-scale fungus strain pleur-jn 21-2F1 full of hyphae into the soil, enriching heavy metals into mushroom bodies and the culture medium along with growth of the thalli, and finally removing the heavy metals by harvesting the culture medium and the thalli.

4. The use of claim 2, wherein when repairing cadmium-contaminated water, the repair is effected by adding hyphae of the macrofungal strain pleur-jn 21-2F1 to the water.