CN106613557A - Microbial control method for plant southern blight - Google Patents

Abstract

The invention relates to the technical field of functional microorganism application, and specifically provides a method for preventing and controlling microorganisms of plant canker sores. The method is to realize the effective prevention and treatment of the canker sore by applying the Penicillium bailei liquid in the soil and/or on the root of the plant and/or on the whole plant. The Penicillium bilaji VLD‑11 ( Penicillium bilaji VLD‑11) was deposited in the Chinese Center for Type Culture Collection of Wuhan University in Wuhan, China on November 23, 2015, and the preservation number is CCTCC NO: M2015690.

Description

A kind of microbial control method of plant canker sore

technical field

The invention belongs to the technical field of microorganism application, and in particular relates to a method for preventing and controlling microorganisms of plant canker sores.

Background technique

Scleritium rolfsii, also known as sclerotinia root rot and sclerotia seedling blight, is a common plant disease in tropical and subtropical regions. Since 1891, Florida State of the United States first reported tomato white spot disease (southern wilt), research reports on various plant spot spot diseases have always been a hot issue in plant disease research.

The fungus can damage many kinds of plants, but the symptoms of the susceptible plants are basically similar. The disease mainly occurs in the rhizomes of seedlings near the ground. When it first occurs, the cortex of the diseased part turns brown, and gradually develops to the surrounding areas. White filamentous mycelium is produced on the lesion, and the mycelium mostly expands radially and spreads to the nearby soil surface. After the disease of the plant, the base of the stem and the cortex of the root rot, the transportation of water and nutrients is blocked, the leaves turn yellow and wither, and the whole plant dies. The dead rhizomes only have lignofibrous tissue, which looks like "messy hemp" and can be easily pulled out from the soil. When the soil is relatively dry, the diseased part is gray-white and dry, and sometimes a white mold layer can be seen; when it is wet, the diseased part is covered with white mycelium, and even the surrounding ground is covered with a layer of mycelium, forming a rapeseed shape Sclerotia.

The asexual stage of the pathogen of white silkworm is Sclerotium Tode ex Fr. of Mycelia Sterilia (Sclerotium Tode ex Fr.), which is a very saprophytic soil. Inhabitant bacteria. The sexual stage of the pathogen belongs to Basidiomycotina, Corticium rolfsii (Sacc.) Curzi.]. The disease rarely occurs under natural conditions, and only in a humid and hot environment, basidiocarps and basidiospores are occasionally seen on the edge of the lesion.

For soil-borne diseases, crop rotation is an important control measure. Reasonable crop rotation, prohibiting continuous cropping or crop rotation with other susceptible hosts, crop rotation with gramineous crops, or paddy and dry crop rotation is very effective in preventing the disease. The study found that the rotation of peanuts and Paspalum americana has a good control effect on white silkworm. In the long-term production practice, the drug farmers also realized that crop rotation with corn, sweet potato, rape and other crops for more than 3 years can effectively prevent and control white silkworm.

In terms of chemical control, Liu Huiqing et al. found through indoor drug efficacy testing that tuzet, kefut, formaldehyde, and mancozeb had a strong inhibitory effect on the germination of sclerotia, and carbendazim, thiophanate-methyl, and chlorothalonil Also has certain effect. The results of further tests on the medicaments showed that 50% carbendazim, 75% chlorothalonil, and 36% formaldehyde had obvious bacteriostatic zones (inhibition of hyphae growth).

In terms of biological control, many reports suggest that Trichoderma has a good effect on the bacterial infection. He Kejia used Trichoderma viride to control white silk disease of pepper, with a control effect of 69%~72%; against konjac white silk disease, the control effect was 65.1%~69.6%. Another useful biocontrol fungus is Viridans viridans, which have been found to be effective against tomato, sweet pepper, potato and pepper blight. Pseudomonas fluorescens can inhibit the occurrence of soybean blight and cotton and potato wilt.

Silkworm is a pervasive and devastating disease with a wide range of hosts, and serious damage can lead to severe crop yield reduction. Biocontrol is a promising field due to the growing concern about environmental issues and the demand for green food. Therefore, there is an urgent need to screen out highly efficient, environmentally friendly and safe biocontrol bacteria to control the incidence and harm of canker sores, so as to achieve good economic and social benefits.

Contents of the invention

In order to solve the problems in the prior art, the present invention provides a microbial control method for canker sores. The method is to inhibit the reproduction of Sclerotinia sclerotiorum by applying Penicillium bilaji , so as to achieve the effect of preventing and treating the white silk disease.

One aspect of the present invention provides a method for preventing and controlling plant blight, which is realized by applying Penicillium bailei fungus liquid in soil and/or on plant roots and/or on whole plants.

The control method of described plant canker sore, comprising:

(1) Before planting or planting, spray Penicillium bayley liquid evenly on the soil at an amount of 40-60mL/m ² , and effectively mix the soil with a thickness of 10-20cm on the surface;

(2) After the plants are planted, apply the penicillium bacterium liquid to the roots of the plants at an amount of 100-500mL/plant, and then apply the same amount again after 15-30 days, and use it 3-5 times in a row;

(3) At the early stage of the onset of white silkworm, spray Penicillium bailei liquid evenly on the whole plant and soil at the dosage of 50-100mL/m ² , and then spray again at the same dosage after 5-10 days; Second-rate.

Further, the Penicillium bilaji is preferably Penicillium bilaji VLD-11 ( Penicillium bilaji VLD-11), which was screened from the soil of the Pengjia Phosphate Mine Area in Laizhou City, Shandong Province, and was preserved in Wuhan, Wuhan, China on November 23, 2015. The Chinese Type Culture Collection Center of the University, the collection number is CCTCC NO: M2015690.

The spore content in the Penicillium bayley liquid is not less than 10 ⁶ CFU/mL.

The plant is preferably any one of carrots, peanuts and jasmine.

The preparation method of the Penicillium bailei bacteria liquid is specifically: first inoculate the above-mentioned Penicillium bailei VLD-11 on the PDA solid medium, cultivate at 30°C for 3-4 days until the spores are mature, and use an appropriate amount of spores After washing with sterile normal saline, transfer to potato liquid culture medium, mix thoroughly and evenly, and cultivate at 30°C for 4-6 days to obtain Penicillium bailei liquid.

The method provided by the invention is to realize the prevention and treatment of plant canker sores by applying the penicillium bayley liquid. The inhibition rates of the Penicillium bayley VLD-11 on Sclerotinia delphinium and Sclerotinia sclerotium reach 85.9% and 82.6% respectively, and the antibacterial effect is remarkable. The Penicillium balaiella VLD-11 can effectively prevent and control plant canker sores. In the prevention and treatment of carrot canker sores, by spraying the 100-fold dilution of Penicillium barley on the whole plant and the soil, the incidence of carrot canker can be reduced to less than 0.5%, and the control efficiency exceeds 88%. In the prevention and control of peanut canker sore, the soil should be treated with Penicillium bailei first, and then the whole plant and soil should be sprayed reasonably at the early stage of canker sore, which can effectively reduce the disease of peanut canker sore Index, the average control efficiency of 50 times of dilutions to peanut canker sore is up to 83.6%; In the prevention and treatment of jasmine canker sore, by directly pouring described Penicillium bailei VLD-11 at the root of jasmine seedling, can effectively reduce The incidence rate of jasmine canker sore in the later stage, the control efficiency of 100 times dilution to canker sore is higher than 85%, the effect is remarkable.

detailed description

The Penicillium bilaji VLD-11 ( Penicillium bilaji VLD-11) selected in the embodiment of the present invention was screened from the soil of the Pengjia Phosphate Mine Area in Laizhou City, Shandong Province, and was preserved in the China National Institute of Wuhan University, Wuhan, China on November 23, 2015. Typical Culture Collection Center, the deposit number is CCTCC NO: M2015690.

Any commercially available product can be used as the reagent used in the present invention, and the method of the present invention will be described in detail below in conjunction with specific examples.

Embodiment 1 Bayley Penicillium antibacterial experiment

1. Preparation of Penicillium bayley VLD-11 bacterial liquid:

First, inoculate the above-mentioned Penicillium bailei VLD-11 on the PDA solid medium, culture at 30°C for 3-4 days until the spores mature, wash the grown spores with an appropriate amount of sterile saline, and transfer them to the potato liquid medium , mixed well and cultured at 30°C for 4 days, the spore content in the fermentation broth was about 10 ⁹ CFU/mL.

2. Inhibitory effect of Penicillium barley VLD-11 on Sclerotinia 1. Inoculate Penicillium barley VLD-11 on a PDA medium plate and culture at 30°C for 7 days;

2. Take the PDA medium plate that has completed step 1, and use a puncher to get a bacterium cake (biological control bacterium cake) with a diameter of 5mm at the front of the colony; Inoculate to PDA medium plate, and cultivate at 30°C for 7 days; 4. Take the PDA medium plate that has completed step 3, and use a puncher to get a bacterium cake (pathogenic bacterium cake) with a diameter of 5 mm at the front of the colony; 5. Take a new one The PDA medium plate of the PDA culture medium plate, and the medium with a diameter of 5 mm was punched out with a puncher as a control. 6. Group treatment (1) Test group: Take a new PDA medium plate, place 1 biocontrol cake and 1 pathogen cake on it, the linear distance between the centers of the two cakes is 3cm, and culture at 30°C; (2) Control group: Take a new PDA medium plate, place a control object and a pathogenic bacteria cake on it, the linear distance between the centers of the two is 3cm, and culture at 30°C. 7. Measure the colony diameter of pathogenic bacteria after 3 days of culture, and calculate the inhibition rate.

Inhibition rate = (colony diameter of pathogenic bacteria in control group - colony diameter of pathogenic bacteria in test group) ÷ colony diameter of pathogenic bacteria in control group × 100%.

The result shows: the inhibitory rate of Penicillium bailei VLD-11 reaches 85.9% and 82.6% (average value repeated three times) to Sclerotinia delphinium and neat Sclerotinia respectively, illustrates that the Penicillium bailei VLD-11 is to The growth and reproduction of Sclerotinia sclerotiorum has a strong inhibitory effect, and it is expected to be used in the prevention and treatment of canker sores.

Example 2 The control effect of Penicillium bailei on carrot mildew

1. Experimental site: Carrot disease area in Pingdu Nancun Town, Qingdao

2. Experimental method

Select three treatment groups, each treatment group has 3 ridges of carrots, and each ridge has about 100 carrots, and a protection row is set up between the treatment groups. Dilute the Penicillium bailei VLD-11 fermentation broth by adding water to 100 times, 200 times and 400 times respectively. In the initial stage of the onset of carrots, use an electrostatic sprayer to fully spray Penicillium bailei liquid on the whole plant and soil at a dosage of 50-100mL/m ² , and then spray with the same dosage once every 5 days, for a total of 3 times of control. The control group was sprayed with the same amount of water. 6 days after the last treatment, check the treatment of carrot blight.

3. Analysis of results

The formula for calculating the control efficiency = [1 - (the diseased plant rate of the treatment / the diseased plant rate of the control)] × 100%

The results are shown in Table 1:

Table 1 Control effect of Penicillium balaiella VLD-11 on carrot mildew

As can be seen from the data in Table 1, at the initial stage of the onset of white silkworm of carrot, by spraying Penicillium bailei VLD-11 to the whole plant and soil, the incidence rate of white silkworm can be significantly suppressed, and 100 times of fermentation broth dilution can The incidence rate of carrot mildew is reduced to less than 0.5%, and the control efficiency exceeds 88%. The effect is remarkable.

Example 3 Control effect of Penicillium bailei VLD-11 on peanut scurvy

1. Experimental location: Peanut planting disease area in Jiaonan City, Qingdao

2. Experimental method:

Dilute Penicillium bailei VLD-11 fermented liquid with water to 50 times, 100 times, 200 times respectively, at the peanut sowing stage, spray Penicillium bailei liquid evenly on the soil at the amount of ^40-60mL /m2, and Effectively mix the 10-20cm thick soil on the surface, and then sow; in the early stage of peanut white silk disease, use an electrostatic sprayer to fully spray the Penicillium ^bailei liquid on the whole plant and the soil at the beginning of the onset of white silk disease. Spray, and then spray once at the same amount every 5 days, for a total of 3 times. 6 days after the last control, check the treatment of peanut canker sores. The control group used clean water.

3. Result analysis:

Disease index = [∑ (disease level value × diseased tree at this level) / (total number of diseased plants × highest disease level value)] × 100%

Control efficiency = [1-(treated disease index/control disease index)] × 100%

The results are shown in Table 2:

Table 2 Control effect of Penicillium baleyi VLD-11 on peanut mildew

As can be seen from the data in Table 2, when peanuts are sown, the Penicillium bailei VLD-11 provided by the present invention is first utilized to treat the soil, and at the initial stage of the disease, the whole plant and the soil are then rationally sprayed. It can effectively reduce the disease index of peanut canker sores. The average control efficiency of Penicillium bailei VLD-11 fermentation broth was 50 times higher than that of peanut canker sores, which was remarkable.

Example 4 The control effect of Penicillium bailei VLD-11 on jasmine mildew

1. Experimental location:

Jasmine Garden, Kutao Flower Market, Laoshan District, Qingdao City.

2. Experimental settings

Three treatment groups were set up, and the fermentation liquid of Penicillium barley VLD-11 was diluted with water by 100 times, 200 times and 400 times respectively.

At the beginning of April 2016, after transplanting jasmine seedlings, the dilution of Penicillium barley VLD-11 was directly applied to the roots of the flower seedlings at an amount of 300 mL/plant, and then applied again 20 days later, for a total of 4 times. Each treatment group was replicated 3 times, with 50 jasmine plants in each replicate. The control group was given water.

3. Statistics on the control effect of jasmine silkworm

At the beginning of November 2016, the incidence of jasmine in each treatment group was investigated, and the control effect of jasmine silkworm was calculated.

Disease grading standards: Grade 0 - no diseased leaves and branches; Grade 1 - the proportion of infected branches and leaves is less than 25%; Grade 3 - the proportion of infected branches and leaves is ≥ 25%, but ≤ 50%; Grade 5 - the proportion of infected branches and leaves is less than 25% >50%, but the plant is not dead; grade 7 - the whole plant is dead.

Disease index = [∑ (disease level value × diseased tree at this level) / (total number of diseased plants × highest disease level value)] × 100%;

Control efficiency = [1-(treated disease index/control disease index)] × 100%;

The results are shown in Table 3:

Table 3 Control effect of Penicillium balaiella VLD-11 on jasmine mildew

As can be seen from the results in Table 3, the Penicillium bailei VLD-11 provided by the present invention can be effectively reduced by directly irrigating the roots of jasmine seedlings, which can effectively reduce the incidence of jasmine mildew in the later stage. The control efficiency of the diluent against the canker sore is higher than 85%, and the effect is remarkable.

In addition to carrots, peanuts and jasmine, the Penicillium bailei VLD-11 provided by the invention can also have significant control effects on plants such as peppers, scallions, walnuts, masson pine, apples, citrus, etc., and the control efficiency reaches More than 80%, broad application prospects.

Claims (6)

1. a control method of plant canker sore, is characterized in that, described method realizes by in soil and/or at plant root and/or at plant whole plant application penicillium balayae. 2. The control method according to claim 1, characterized in that, said method comprises: (1) Before planting or planting, spray Penicillium bayley liquid evenly on the soil at an amount of 40-60mL/m ² , and effectively mix the soil with a thickness of 10-20cm on the surface; (2) Irrigate the Penicillium barley fungus liquid on the root of the plant according to the dosage of 100-500mL/plant, and then apply it again with the same dosage after 15-30 days, and use it 3-5 times in a row; (3) Spray Penicillium barley liquid evenly on the whole plant and soil at the dosage of 50-100mL/m ² , and then spray again at the same dosage after 5-10 days; use 3-5 times in succession. 3. The control method according to claim 1 or 2, characterized in that, the preservation number of said Penicillium bailei is CCTCCNO: M2015690. 4. The control method according to claim 1 or 2, characterized in that the spore content in the Penicillium bayley liquid is not less than 10 ⁶ CFU/mL. 5. The control method according to claim 1 or 2, wherein the plant is any one of carrots, peanuts and jasmine. 6. The control method according to claim 1 or 2, characterized in that, the preparation method of the Penicillium bailei liquid is specifically: first inoculate the Penicillium bailei on the PDA solid medium, and cultivate it at 30°C After 3-4 days until the spores mature, wash the grown spores with an appropriate amount of sterile saline, transfer them into potato liquid medium, mix them well, and culture them at 30°C for 4-6 days to obtain Penicillium bailei liquid.