CN101965848B - Biogenic pesticide for efficiently preventing and controlling main diseases of Camellia oleifera and application thereof - Google Patents

Abstract

The invention discloses a biogenic pesticide for efficiently preventing and controlling main diseases of camellia oleifera (anthracnose, soft rot and leaf blight of the camellia oleifera), namely a pesticidal aqueous solution agent compounded by an clove extract, a skullcap extract, an orange peel extract, tea saponin and bacterial filtrate after bacillus subtilis Y13 is fermented, also discloses a preparation method and application thereof, and belongs to the technical field of plant protection. The biogenic compounded medicine can effectively inhibit the development of the anthracnose, soft rot and leaf blight of the camellia oleifera, improve the resistance of the camellia oleifera to the anthracnose, soft rot and leaf blight and efficiently prevent and control the main diseases of the camellia oleifera, has high environmental safety and has good development and application prospect.

Description

A kind of biological source pesticide and its application of high-efficiency prevention and treatment of major diseases of camellia oleifera

technical field

The invention belongs to the field of plant protection, and in particular relates to a biological source pesticide for preventing and treating anthracnose, soft rot and leaf blight of camellia oleifera and its application.

Background technique

Camellia oleifera (Camellia oleifera) is a unique edible oil tree species in my country and one of the four largest woody oil tree species in the world. It has a history of utilization and cultivation for more than 2,000 years. It has wide adaptability and strong stress resistance, and is suitable for planting in vast red soil hilly areas. It mainly grows in the south, mainly in Hunan, Jiangxi, and Guangxi. Among them, Hunan Province is the largest camellia oleifera producing area and the largest camellia oil consumption area in China. Camellia oleifera is full of treasures and has high economic value. The unsaturated fatty acid content of camellia oil is as high as 90%, which is much higher than that of rapeseed oil, peanut oil and soybean oil. It is twice as high as olive oil in vitamin E content, and contains specific physiologically active substances such as camellia glucoside, which has extremely high nutritional value. According to statistics, China now produces 130 to 150 million kilograms of camellia oil per year, which is the main source of edible oil for a population of more than 200 million. However, there are many kinds of diseases of camellia oleifera, and the degree of harm is becoming more and more serious. The annual economic losses caused by diseases are difficult to estimate, and the economic losses caused by camellia anthracnose, soft rot and leaf blight are particularly serious.

Camellia anthracnose, soft rot and leaf blight are caused by the infection of Colletotrichum gloeosporioides, Agaricodochium camellia Liu, Wei et Fan and Pestalotiopsis microspora respectively The three devastating diseases are the main diseases in Camellia oleifera producing areas in my country, and they occur widely. When the diseases are severe, they will cause serious fruit drop, bud drop, leaf fall, branch withering, and even the whole plant decline. The disease fruit drop rate is usually about 20%, and more than 40% in severe cases. At present, the prevention and treatment of anthracnose and soft rot of Camellia oleifera is mainly controlled by planting resistant varieties and chemical control, but excessive use of chemical fungicides will endanger human health, destroy the ecological environment, and easily cause pathogenic bacteria to develop drug resistance, affecting tea. Oil quality is not conducive to the sustainable development of the entire Camellia oleifera industry. The search for safe, low-toxicity, and high-efficiency biogenic pesticides has become a common concern of the society in the prevention and control of Camellia oleifera pests and diseases. Due to the characteristics of high efficiency, low toxicity and low residue of biological source pesticides, searching for bactericidal and antibacterial active substances from plants, insects and microorganisms has become one of the hot spots in the development and development of new pollution-free biological source fungicides. Its main advantages are: 1) There are many types and quantities of organisms, especially plants and microorganisms, and they have good environmental compatibility. Biogenic pesticides have a smooth degradation pathway in nature and have little environmental pollution; 2) Various biological activities. The extracts and secretions of plants themselves, microorganisms and their metabolites not only have bactericidal activity, but also have the functions of killing insects and regulating plant growth, and their modes of action are diverse; 3) Safe for higher animals and natural enemies of pests; 4) Harmful organisms Not easy to develop drug resistance. Biological pesticides often contain several active ingredients, and the mechanism of action is different from that of general chemical pesticides, so it is not easy to make harmful organisms resistant to pesticides; 5) It is safe for agricultural and forestry crops, and some secretions, microorganisms and metabolites can induce agricultural and forestry crops to produce Anti-disease effect;

Contents of the invention

The purpose of the present invention is to aim at the gap in the prior art, to provide a kind of biological source pesticide which can efficiently prevent and treat major diseases of camellia oleifera including anthracnose, soft rot, leaf blight, etc., which is of great significance to the pollution-free prevention and treatment of camellia oleifera.

The purpose of the present invention is achieved in the following manner:

A biogenic pesticide for effectively preventing and treating major diseases of Camellia oleifera is prepared by mixing plant extract compound liquid A with bacterial filtrate B fermented by Bacillus subtilis Y13 at a volume ratio of 1-10:1-10; The plant extract compound solution A is a water solvent including the following components in mass content: clove extract 5%-10%, scutellaria baicalensis extract 5%-10%, orange peel extract 5%-15%, tea saponin 5 -10%; the Bacillus subtilis Y13 preservation number CCTCC NO.M208264 (this bacterial strain has applied for a patent for invention, and is a kind of endophytic bacterium isolated from the healthy leaves of Camellia oleifera by the Forest Protection Laboratory of Central South University of Forestry and Technology) .

The extraction process of the clove extract, scutellaria baicalensis and orange peel extracts in the plant source compound solution A is as follows: take 60 meshes of dried cloves, scutellaria baicalensis and orange peel powder at a temperature of 70°C to 90°C, and use cloves, Scutellaria baicalensis and orange peel powder weight 10-30 times the volume of sterile water, water bath extraction at least twice, each water bath extraction time is 3-5 hours, after each extraction, filter, finally combine the filtrate, concentrate under reduced pressure to the water content About 4-6% paste, to obtain clove extract, scutellaria baicalensis extract and orange peel extract respectively.

The purity of the tea saponin is 70%-95%.

The bacterial filtrate B after the fermentation of Bacillus subtilis Y13 is to cultivate Y13 on the NA medium in a constant temperature incubator at 28°C for 2 days, then use the inoculation loop to pipette a loop into 150ml of NB In the Erlenmeyer flask of the culture solution, shake culture on a shaker at 28° C. and 150 rpm for 2-4 days; the culture solution is filtered with a 0.22 μm bacterial filter to obtain a bacterial filtrate after fermentation.

In the preparation method of the biological source pesticide, the required tea saponin is firstly weighed and dissolved in sterile water at a temperature of 35-60° C., and then the clove extract, the scutellaria baicalensis extract and the orange peel extract are weighed respectively by weight, Prepare plant extract compound solution A; then mix the prepared plant extract compound solution with Bacillus subtilis Y13 fermented bacterial filtrate B at a volume ratio of 1-10:1-10.

The biological source pesticide is applied to the prevention and treatment of Camellia oleifera diseases such as anthracnose, soft rot and leaf blight in forests. The present invention has following beneficial effect:

1) The required plants in the biogenic pesticide of the present invention are widely distributed in China and have many resources. At the same time, the present invention also mixes the fermented bacterial filtrate of Camellia oleifera endophytic bacteria Bacillus subtilis Y13 with the preservation number CCTCC NO.M208264 and the plant extract, which can further promote the absorption of camellia oleifera to the biological source pesticide of the present invention, and does not pollute the environment. .

2) The biological source pesticide of the present invention can not only efficiently prevent and treat camellia oleifera diseases in the forest, but also promote the growth of camellia oleifera, induce the camellia oleifera to produce resistance to anthracnose, soft rot, and leaf blight, so that the dosage of the medicament is reduced and the drug effect is improved. improve.

3) The biological source pesticide of the present invention has the advantages of simple manufacturing process, little environmental pollution, no pollution and no residual toxicity, and effectively avoids the harm of chemical pesticides.

4) The biogenic pesticide of the present invention, on the one hand, through the compounding of plant extracts and the mixing with the bacterial filtrate after fermentation of Y13, greatly reduces the possibility of Camellia oleifera pathogenic bacteria producing resistance to the biogenic pesticide of the present invention , on the other hand, the economic cost of single drug forest control is greatly reduced by mixing, and the control effect is good.

Detailed ways

The present invention will be further described in detail below in conjunction with the examples, without limiting the present invention.

Example 1

Screening of plant extracts for effective control of major diseases of camellia oleifera

Using mycelia growth rate method: respectively draw 1ml of the plant extract solution with a concentration of 20mg/ml described in Table 1, and pour it into a sterilized Petri dish containing 9ml of PDA, and use 1ml of sterile water for the control, and wait for the culture medium After solidification, use a puncher with a diameter of 0.8 cm to make a fungus dish, use an inoculation needle to inoculate the fungus dish to the center of the culture medium, with the mycelium facing down, one piece for each dish, repeat 3 times for each treatment, and culture in a 28 incubator , and the colony diameter was measured by the cross method after 4 days of culture. Inhibition rates were calculated as the average of three replicates. The results are shown in Table 1

Table 1 Inhibitory effect of different plant extracts on major diseases of camellia oleifera

Example 2

Effects of Biogenic Pesticides with Different Volume Ratio on the Growth of Camellia oleifera

Select 2-year-old Camellia oleifera seedlings with the same growth, and measure the height of each seedling, spray different proportions of biological source pesticides on the seedlings, and use sterile water as a control, spray 20 seedlings for each treatment, and spray again after 7 days. Measure the height of the seedlings one month later. The influence of biogenic pesticides on the growth of camellia oleifera was measured, and the results are shown in Table 2. Biogenic pesticides with different volume ratios can all promote the growth of camellia oleifera to a certain extent.

Table 2 Effects of different volume ratios of biogenic pesticides on the growth of Camellia oleifera

Note: A ₁ is a compound solution of 5% clove extract + 5% scutellaria baicalensis extract + 5% orange peel extract + 5% tea saponin plant extract.

A ₂ is a compound solution of 10% clove extract + 10% scutellaria baicalensis extract + 10% orange peel extract + 10% tea saponin plant extract.

B is the bacterial filtrate after fermentation of Bacillus subtilis Y13 (B is after culturing the Y13 strain on the NA medium for 2 days, pipette a ring with the inoculation loop and put it into a conical flask containing 150ml NB culture solution, and put it in a flask at 28°C , 150rpm shaking on a shaker for 2-4 days; the culture solution is filtered with a 0.22 μm bacterial filter to obtain a bacterial filtrate after fermentation).

C ₁ is a compound solution of 20% clove extract + 10% tea saponin plant extract

C ₂ is a compound solution of 20% Scutellaria baicalensis extract + 10% tea saponin plant extract

The same letters in the table below represent the same meaning.

Example 3

Induced and resistive effects of biogenic pesticides with different volume ratios on camellia oleifera

After mixing the biogenic pesticides according to the volume ratio in Table 3, spray the liquid medicine on the 2-year-old Camellia oleifera seedlings until water drips from the leaves, and then spray the liquid medicine again after 2 days. The yellow-green young leaves that had been sprayed with the liquid medicine were collected and inoculated, and distilled water was used as a control. Each treatment had 15 replicates. Observation results. It can be seen from Table 3 that the mixed biogenic pesticides with different volume ratios can induce the resistance of camellia oleifera to the main diseases of camellia oleifera.

Lesion reduction rate = lesion area of control treatment - lesion area of induction treatment / lesion area of control treatment × 100%

Table 3 Induction and resistance effects of different proportions of biogenic pesticides on camellia oleifera

Example 4

Forest control efficacy test of biogenic pesticides with different volume ratios on major diseases of camellia oleifera

1. Test plants and control objects

Test plants: 2-year-old Camellia oleifera seedlings

Control objects: Camellia anthracnose, soft rot, leaf blight

2. Drugs to be tested

Different proportions of biogenic pesticide, carbendazim (Changsha Pesticide Co., Ltd.)

3. Test method

Camellia oleifera is a natural disease. After mixing biogenic pesticides according to different volume ratios, 2-year-old seedlings with the same growth were selected in early April and sprayed first, and sprayed again after 5 days. At the same time, sterile water and standard pesticides were used as controls. 15 seedlings constitute one treatment, and each treatment has 3 replicates. After 30 days, the condition investigation was carried out. According to the grading standard of camellia oleifera anthracnose, the disease index was counted, as shown in Table 4, and the preventive effect was calculated. The standards and formulas are as follows:

Disease index = ∑ (number of plants at the disease level × representative value of the disease level) / (number of investigated plants × representative value of the highest disease level)

Control effect (%)=(average disease index of the control group-average disease index of the application group)/average disease index of the pair group×100

Table 4 Criteria for grading diseases of camellia oleifera

4. Experimental results

Table 5 The results of the control efficacy test of biogenic pesticides in forests with different volume ratios

It can be seen from Table 5 that within the volume ratio range of 1-10:1-10, the biological source pesticides of the present invention can effectively prevent and treat diseases of camellia oleifera.

Claims (5)

1. A biologically derived pesticide for efficiently preventing and treating major diseases of camellia oleifera, characterized in that it is made of plant extract compound solution A and Bacillus subtilis Y13 fermented bacterial filtrate B in different volume ratios of 1-10:1-10 Formed by mixing; the plant extract compound solution A is a water solvent comprising the following components by mass content: clove extract 5%-10%, scutellaria baicalensis extract 5%-10%, orange peel extract 5% -15%, tea saponin is 5%-10%; said Bacillus subtilis Y13 preservation number CCTCCNO.M208264; The extraction process of the clove extract, scutellaria baicalensis and orange peel extracts in the plant source compound solution A is as follows: take 60 meshes of dried cloves, scutellaria baicalensis, and orange peel powders at a temperature of 70°C to 90°C, and use cloves, Scutellaria baicalensis and orange peel powder weight 10-30 times the volume of sterile water, water bath extraction at least twice, each water bath extraction time is 3-5 hours, after each extraction, filter, finally combine the filtrate, concentrate under reduced pressure to the water content It is about 4-6% paste, and clove extract, scutellaria baicalensis extract and orange peel extract are obtained respectively. 2. The biogenic pesticide for efficiently preventing and treating major diseases of Camellia oleifera according to claim 1, wherein the purity of the tea saponin is 70%-95%. 3. the biological source pesticide of the efficient prevention and treatment of Camellia oleifera main disease according to claim 1, it is characterized in that, the bacterial filtrate B after the fermentation of described Bacillus subtilis Y13 will be on the Y13 bacterial strain of NA substratum at 28 After cultivating in a constant temperature bacterial incubator at ℃ for 2 days, pipette a ring with an inoculation loop and put it into a conical flask containing 150ml of NB culture solution, and shake it on a shaking table at 28℃ and 150rpm for 2-4 days; the culture solution Filtrate with a 0.22 μm bacterial filter to obtain the post-fermentation bacterial filtrate. 4. according to the preparation method of claim 1 or 2 or 3 described biogenic pesticides, it is characterized in that, first take required tea saponin and be dissolved in the aseptic water of 35-60 ℃ of temperature, then weigh respectively by weight Take clove extract, scutellaria baicalensis extract, and orange peel extract to make plant extract compound solution A; then prepare plant extract compound solution A and Bacillus subtilis Y13 fermented bacterial filter solution B, by volume Ratio 1-10: 1-10 after mixing. 5. The biological source pesticide described in claim 1 or 2 or 3 is used for preventing and treating anthracnose of Camellia oleifera, soft rot and leaf blight of Camellia oleifera.