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Biological agent for promoting plant growth and controlling diseases and pests

A technology for biological preparations and strains, applied in the directions of plant growth regulators, plant growth regulators, and chemicals for biological control, which can solve difficult large-scale applications, reduce biological activity and stability, reduce microbial protein and Peptide yield and other issues

Active Publication Date: 2017-11-17
湖北微生元生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are non-chemical methods for cell lysis, such as high pressure or ultrasonic treatment, which require high equipment requirements, high energy consumption and are difficult to apply on a large scale
As an alternative, most manufacturers use the method of contacting the cell suspension with lysozyme, followed by digestion at 40-42°C, centrifugation to remove cell debris and denatured proteins, and protein purification processes. Reduces the yield of microbial proteins and peptides and reduces their biological activity and stability

Method used

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  • Biological agent for promoting plant growth and controlling diseases and pests
  • Biological agent for promoting plant growth and controlling diseases and pests
  • Biological agent for promoting plant growth and controlling diseases and pests

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] Embodiment 1, the production of Harpin biological preparation stoste

[0092]Take one strain from the working seed bank, put it into a 200ml Erlenmeyer flask filled with 50ml LB medium (containing 34μg / ml kanamycin) after melting, culture at 37°C, 100-200rpm / min, for 6-8h 1ml was added to 20 Erlenmeyer flasks with 2000ml capacity containing 500ml LB medium (containing 34μg / ml kanamycin), cultivated overnight at 37°C, 100-200rpm / min, to obtain amplified 10 liters of bacteria solution.

[0093] 200 liters of basal medium preparation: 1000 grams of peptone, 500 grams of yeast extract, 820 grams of anhydrous sodium phosphate, 1080 grams of sodium dihydrogen phosphate, 260 grams of ammonium sulfate, 40 grams of ammonium chloride, 600 grams of glycerin, 200 liters of purified water , 200ml silicone oil (Lanxiang Chemical), sterilized at 121°C for 20 minutes.

[0094] Magnesium sulfate solution preparation: 110 grams were dissolved in 1.8 liters of purified water, and steril...

Embodiment 2

[0107] Purification of embodiment 2 Harpin protein and assay method of stoste function

[0108] Detection of biological activity of Harpin biologics,

[0109] a. Purification of Harpin protein

[0110] Take 10ml of the bacteria solution after induced expression, centrifuge at 12000RPM for 3min, discard the supernatant, weigh the precipitate, resuspend the bacteria in 20mM TrisHCl (pH8.0) according to the ratio of 5mg / 100ul, and ultrasonically break; The supernatant; use 5 column volumes of 20mM TrisHCl (pH8.0) to equilibrate the nickel column, then use the supernatant after the previous step of centrifugation to pass through the nickel column, and then use the washing buffer (20mM TrisHCl, pH8.0, 20mM imidazole) Wash 5 column volumes, wash 5 column volumes with elution buffer (20mM TrisHCl, pH8.0, 250mM imidazole), the results are as follows figure 1 As shown, the Harpin protein with a purity of more than 95% was obtained, and after quantification, it was freeze-dried and s...

Embodiment 3

[0129] Embodiment 3, Harpin biological agent foliar application effect

[0130] Take fruit trees, tobacco, tea and other crops, spray the Harpin biological agent stock solution prepared in Example 1 at the seedling stage and rapid growth stage of the crops, and spray the leaves of the crops 3-4 times in total, and spray at a ratio of 1:500 at the seedling stage 1:1000 ratio in other periods;

[0131] Tobacco spray effects such as figure 2 As shown, compared with the control group (only containing the same concentration of urea, without harpin protein), the leaves are obviously thicker and larger, the plants are also taller, and the statistical results show that the dry matter increases by more than 20%.

[0132] The effect of pumpkin spraying is as follows image 3 As shown, it can be seen that the germination rate increased by about 60% after spraying compared to the control group that failed to spray.

[0133] The spraying effect of netted melon is as follows: Figure 4...

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Abstract

The invention relates to a method for preparing a biological agent containing allergenic protein (Harpin). The method is characterized by comprising the following steps: (1) recovery and amplification of strains: taking the working seed bank strains transformed with the allergenic protein (Harpin), and performing thawing and multiplication culture; (2) fermentation in a fermentation tank: 1) performing strain domestication, 2) performing thallus amplification and 3) performing induction expression; (3) treatment of a fermentation broth: slowly adding urea to the fermentation broth until the final urea concentration is 8 mol / L and urea is completely dissolved, continuing stirring for 15-45 minutes, and cooling the liquid composition to 2-8 DEG C.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a biological preparation for promoting plant growth and preventing and controlling diseases and insect pests. Background technique [0002] Chemical fertilizers and pesticides are the main products to increase crop yields and prevent pests and diseases, but at the same time, they have caused problems in the environment, energy and cost, and have become a major issue that plagues the economic and social development of countries around the world and seriously restricts the sustainability of agriculture. develop. The use of biological products (including biological fertilizers and biological pesticides) prepared from proteins from microorganisms or fungi is the main measure for high-quality, efficient and pollution-free production of contemporary crops. In recent years, a lot of scientific research has been carried out in this area at home and abroad. The early microbial pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/02C12N15/70A01N63/02A01N47/44A01P21/00A01P3/00A01P7/04C05G3/00C12R1/19
CPCA01N47/44A01N63/10C05F11/08C05G3/00C05G5/20C07K14/195C07K14/21C07K14/27C12N15/70A01N2300/00
Inventor 刘欣胡业勤刘军鹏刘胜朱德武
Owner 湖北微生元生物科技有限公司
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