阅读说明：本技术 一种植物保护剂的制备及其使用方法 (Preparation and application method of plant protective agent ) 是由 周廷霖 于 2019-09-18 设计创作，主要内容包括：本发明涉及一种植物保护剂的制备及其使用方法,属于植物保护试剂技术领域。该植物保护剂的制备以利迪链霉菌、蔗糖、酵母提取物、硫酸镁、磷酸二氢钾和氯化锰为主要原料制备而成。本发明中的利迪链霉菌植物保护剂,是以利迪链霉菌株发酵液,确认每毫升发酵液营养细胞浓度至10^7 cfu/ml以上后,接种至以煮熟之大米为主要载体上,经过48小时恒温培养,再经低温40度干燥水分至10%以下制成。该植物保护剂的制备方法工艺设计科学合理,制备成本低,工艺简单易控制,适合推广使用。(The invention relates to a preparation method and a use method of a plant protective agent, belonging to the technical field of plant protective agents. The plant protectant is prepared by taking streptomyces lydicus, cane sugar, yeast extract, magnesium sulfate, monopotassium phosphate and manganese chloride as main raw materials. The streptomyces lydicus plant protective agent is prepared by using streptomyces lydicus strain fermentation liquor, determining that the nutrient cell concentration of each milliliter of the fermentation liquor is more than 10^7 cfu/ml, then inoculating the fermentation liquor to a boiled rice serving as a main carrier, carrying out constant temperature culture for 48 hours, and drying at the low temperature of 40 ℃ until the water content is less than 10%. The preparation method of the plant protectant has the advantages of scientific and reasonable process design, low preparation cost, simple and easily controlled process and suitability for popularization and use.)

1. A preparation method of a plant protective agent is characterized by comprising the following steps:

(1) firstly, culturing streptomyces lydicus in a liquid shake flask, taking 10 percent (weight percentage) of cane sugar, 5 percent (weight percentage) of yeast extract, 0.15 percent (weight percentage) of magnesium sulfate, 0.3 percent (weight percentage) of monopotassium phosphate and 0.02 percent (weight percentage) of manganese chloride as liquid culture media, and carrying out shake flask culture for 7 days at the constant temperature of 30 ℃ and the rotating speed of 150 revolutions per minute so that the concentration of the vegetative cells of the streptomyces lydicus per milliliter reaches more than 10^7 cfu/ml;

(2) before solid fermentation, the materials and the pot are sterilized to ensure the aseptic state, then the rice is washed clean and steamed for more than 1 hour to be cooked and soft under the aseptic state, and then the rice is placed in the pot and cooled to 20 ℃;

(3) after the steamed rice is cooled, grinding the rice to 200 meshes, transferring the rice to a solid stainless steel fermentation tank, and inoculating the streptomyces dikaensis solution prepared in the step (1) with the weight of 10% of the weight of the rice;

(4) introducing sterile air into a stainless steel fermentation tank, wherein the air flow is 20-30 liters per minute, the stirring speed is 120 revolutions per minute, the temperature is controlled at 30 ℃, the pH is controlled between 6.8-7.4, and after 7 days of stirring fermentation, the activity of the bacteria count, the spore formation and the pH value parameter are periodically sampled and detected during the culture and growth period;

(5) and after fermentation, cooling and drying at minus 40 ℃ for 2-4 days until the water content is below 10%, removing the water content, maintaining the high survival rate of the thalli, and packaging into a finished product after drying and cooling, thereby preparing the plant protective agent.

2. The method of using a plant protectant of claim 1, comprising: when the plant protection agent is used, the plant protection agent and water are uniformly mixed according to the weight ratio of 1:500, and the plant protection agent is applied in a root irrigation mode, a hole application mode or a mode of mixing with a culture substrate before and after plant planting, during plant growth or at the early stage of disease occurrence, so that the disease threat of bacterial wilt can be reduced for plants.

Technical Field

The invention relates to a preparation method and a use method of a plant protective agent, belonging to the technical field of plant protective agents.

Background

Due to the increasing severity of soil-borne diseases in recent years, in solanaceae such as tomato, especially, the bacterial wilt is the most serious, which is caused by pathogenic bacteria, Pseudomonas solanacearum, a soil-borne bacterial disease, invades from root or stem base wounds, spreads in vascular bundle tissues in plants, causes vascular bundle blockage, nutrient failure to be delivered, cell poisoning and plant withering and death. Because pathogenic bacteria can be hidden in soil for years, even if the pesticide is used excessively, the pathogenic bacteria cannot be effectively killed, and the excessive use of the pesticide can cause harm to the environment and the self health, so that the natural microorganisms are utilized to inhibit the growth of the pathogenic bacteria in the aspect of bacteria inhibition, and the prevention is superior to the moxibustion mode, so that the harm to crops is avoided, and the fundamental solution is achieved.

Streptomyces is a genus of actinomycetes and is widely distributed in soil, and currently, more than 1000 antibiotics are known to be extracted from streptomyces such as streptomycin and tetracycline, and can be used for producing various substances with bioactive molecules in soil, so that the streptomyces not only can inhibit pathogenic bacteria, but also can protect the root system health of plants and promote the growth of the plants, and plays an important role in soil.

Tomato bacterial wilt is caused by pathogenic bacteria, Pseudomonas solanacearum, is one of the common vascular bundle systemic diseases on tomatoes in the south, can damage various crops such as tomatoes, eggplants, peppers, potatoes, gingers and the like, and is suitable for the attack in a high-temperature and high-humidity soil meta-acid environment. At the early stage of bacterial wilt, the leaves of the bacterial wilt often droop at high temperature at noon, the pseudomonas solanacearum recovers to a normal sleeping lunch phenomenon, and after several days of repetition, the lower leaves of the diseased plant droop, the leaves gradually wither, and then the whole plant withers. The withered plant still presents green color, so the plant is called as bacterial wilt. Fresh disease stems are transversely cut, squeezed or inserted into clear water by hands, milky white bacteria liquid overflows, which is the main characteristic of the disease that the bacterial wilt belongs to soil-borne diseases, can be propagated from roots to roots and also can be attached to shoes, agricultural machinery and equipment of soil and the flowing of irrigation water and also can carry pathogenic bacteria to propagate, the pathogenic bacteria can survive in the soil for a long time for years, the bacterial wilt can be invaded and infected in the soil from natural openings of the roots or wounds caused by transplantation, nematode, insects or other physical injuries, and the like, and if the cutter carries the bacterial wilt and then prunes healthy plants, the wound infection at the pruning position can be used, so huge harm is brought to tomato planting, and more than 3-5 losses are caused to tomato farmers, even the tomato planting is completely harvested.

There is no suitable solution for tomato bacterial wilt, and it is generally recommended to use 81.3% wettable powder of pesticide, Copper oxychloride (Copper oxychloride), which is ineffective, and has respiratory toxicity and toxicity to aquatic animals. Moreover, the application of pesticide can not effectively kill the ralstonia solanacearum in the soil, so the method is a method for treating the symptoms and the root causes.

The invention relates to a streptomyces lydicus with an inhibiting effect screened by using natural nontoxic microorganism bacteria and utilizing a new natural nontoxic disease control mode of a bacterium-making mode, which is directed at pseudomonas solanacearum seriously harming tomatoes. The results of the laboratory antibacterial antagonism test, the small-amount inoculation test and the actual field operation test show that the streptomyces lydicus can effectively reduce the harm of the tomato bacterial wilt. In addition, the streptomyces lydicus has inhibitory effects on various other pathogenic bacteria, such as Rhizoctonia solani (Rhizoctonia solani), Fusarium oxysporum (Fusarium oxysporum), Colletotrichum gloeosporioides (Colletotrichum gloeosporioide), and tomato early blight (Alternaria solani) …, and is a strain with great biological control potential.

Disclosure of Invention

In order to overcome the defects in the background art, the technical scheme adopted by the invention for solving the technical problems is as follows: a preparation method of a plant protective agent is characterized by comprising the following steps:

(1) firstly, culturing streptomyces lydicus in a liquid shake flask, taking 10 percent (weight percentage) of cane sugar, 5 percent (weight percentage) of yeast extract, 0.15 percent (weight percentage) of magnesium sulfate, 0.3 percent (weight percentage) of monopotassium phosphate and 0.02 percent (weight percentage) of manganese chloride as liquid culture media, and carrying out shake flask culture for 7 days at the constant temperature of 30 ℃ and the rotating speed of 150 revolutions per minute so that the concentration of the vegetative cells of the streptomyces lydicus per milliliter reaches more than 10^7 cfu/ml;

(2) before solid fermentation, the materials and the pot are sterilized to ensure the aseptic state, then the rice is washed clean and steamed for more than 1 hour to be cooked and soft under the aseptic state, and then the rice is placed in the pot and cooled to 20 ℃;

(3) after the steamed rice is cooled, grinding the rice to 200 meshes, transferring the rice to a solid stainless steel fermentation tank, and inoculating the streptomyces dikaensis solution prepared in the step (1) with the weight of 10% of the weight of the rice;

(4) introducing sterile air into a stainless steel fermentation tank, wherein the air flow is 20-30 liters per minute, the stirring speed is 120 revolutions per minute, the temperature is controlled at 30 ℃, the pH is controlled between 6.8-7.4, and after 7 days of stirring fermentation, the activity of the bacteria count, the spore formation and the pH value parameter are periodically sampled and detected during the culture and growth period;

(5) and after fermentation, cooling and drying at minus 40 ℃ for 2-4 days until the water content is below 10%, removing the water content, maintaining the high survival rate of the thalli, and packaging into a finished product after drying and cooling, thereby preparing the plant protective agent.

The application method of the plant protective agent is characterized in that the plant protective agent and water are uniformly mixed according to the weight ratio of 1:500 when the plant protective agent is used, and the plant protective agent is applied in a root irrigation mode, a hole application mode or a cultivation substrate mixing mode before and after plant planting, during plant growth or at the early stage of disease occurrence, so that the disease threat of bacterial wilt of plants can be reduced.

The streptomyces lydicus plant protective agent is prepared by using streptomyces lydicus strain fermentation liquor, determining that the nutrient cell concentration of each milliliter of the fermentation liquor is more than 10^7 cfu/ml, then inoculating the fermentation liquor to a boiled rice serving as a main carrier, carrying out constant temperature culture for 48 hours, and drying at the low temperature of 40 ℃ until the water content is less than 10%. The preparation method of the plant protectant has the advantages of scientific and reasonable process design, low preparation cost, simple and easily controlled process and suitability for popularization and use.

Detailed Description

The preparation method of the plant protective agent is characterized by comprising the following steps:

(1) firstly, culturing streptomyces lydicus in a liquid shake flask, taking 10 percent (weight percentage) of cane sugar, 5 percent (weight percentage) of yeast extract, 0.15 percent (weight percentage) of magnesium sulfate, 0.3 percent (weight percentage) of monopotassium phosphate and 0.02 percent (weight percentage) of manganese chloride as liquid culture media, and carrying out shake flask culture for 7 days at the constant temperature of 30 ℃ and the rotating speed of 150 revolutions per minute so that the concentration of the vegetative cells of the streptomyces lydicus per milliliter reaches more than 10^7 cfu/ml;

(2) before solid fermentation, the materials and the pot are sterilized to ensure the aseptic state, then the rice is washed clean and steamed for more than 1 hour to be cooked and soft under the aseptic state, and then the rice is placed in the pot and cooled to 20 ℃;

(3) after the steamed rice is cooled, grinding the rice to 200 meshes, transferring the rice to a solid stainless steel fermentation tank, and inoculating the streptomyces dikaensis solution prepared in the step (1) with the weight of 10% of the weight of the rice;

(4) introducing sterile air into a stainless steel fermentation tank, wherein the air flow is 20-30 liters per minute, the stirring speed is 120 revolutions per minute, the temperature is controlled at 30 ℃, the pH is controlled between 6.8-7.4, and after 7 days of stirring fermentation, the activity of the bacteria count, the spore formation and the pH value parameter are periodically sampled and detected during the culture and growth period;

(5) and after fermentation, cooling and drying at minus 40 ℃ for 2-4 days until the water content is below 10%, removing the water content, maintaining the high survival rate of the thalli, and packaging into a finished product after drying and cooling, thereby preparing the plant protective agent.

When the plant protective agent is used, the plant protective agent and water are uniformly mixed according to the weight ratio of 1:500, and the plant protective agent is applied in a root irrigation mode, a hole application mode or a mode of mixing with a culture substrate before and after plant planting, during plant growth or at the initial stage of disease occurrence, so that the disease threat of bacterial wilt of plants can be reduced.

The streptomyces lydicus plant protective agent is prepared by using streptomyces lydicus strain fermentation liquor, determining that the nutrient cell concentration of each milliliter of the fermentation liquor is more than 10^7 cfu/ml, then inoculating the fermentation liquor to a boiled rice serving as a main carrier, carrying out constant temperature culture for 48 hours, and drying at the low temperature of 40 ℃ until the water content is less than 10%. The preparation method of the plant protectant has the advantages of scientific and reasonable process design, low preparation cost, simple and easily controlled process and suitability for popularization and use.

It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.