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Protein nano complex promoting plant growth and preparation method and application thereof

A technology that promotes plant growth and nanocomposites. It is applied in the field of protein nanocomposite preparation. It can solve the problems of short service life and serious drug resistance of chemical fungicides, and achieve the goal of improving bioavailability, promoting absorption and utilization, and increasing production. Effect

Active Publication Date: 2013-06-19
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the use cycle of chemical fungicides is getting shorter and shorter, leading to more and more resistance to them
In addition, chemical fungicides are powerless against many diseases

Method used

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  • Protein nano complex promoting plant growth and preparation method and application thereof
  • Protein nano complex promoting plant growth and preparation method and application thereof
  • Protein nano complex promoting plant growth and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Embodiment 1: Preparation of Harpin protein

[0053] 1. Purification of Harpin protein

[0054] The engineered strain E.coli BL21 (China Center for Type Culture Collection CCTCC M202026) containing the harpin gene was used to induce the expression of genetically engineered recombinant Harpin protein (rHrpZ protein, rHrpZ). Pick the strains preserved with glycerol, inoculate them in LB liquid medium containing 200 μg / mL ampicillin (1 L of LB liquid medium: 10 g of peptone, 5 g of yeast powder, 10 g of NaCl, dissolve in 800 ml of double distilled water, adjust the pH with NaOH to 7.0, add water to make the volume to 1000ml), 37°C, shake culture at 200r / min for 8-12h. The next day, inoculate 4% of fresh 2YT liquid medium containing 200 μg / mL ampicillin (peptone 16g, yeast powder 10g, NaCl 5g, dissolve in 800ml double distilled water, adjust pH to 7.0 with NaOH, add water to 1000ml ), at 37°C and 200r / min, shake the culture until the OD600 value is about 0.6-0.8, add the ...

Embodiment 2

[0068] Embodiment 2: the preparation of Harpin protein nanocomplex

[0069] Weigh 250mg of PLGA (poly d, l-lactide-co-glycolide, lactide:glycolide=75:25, molecular weight 40,000, product of Shandong Yanling Chemical Co., Ltd.), dissolve it in 5mL of dichloromethane, and make a 50mg / mL solution . Slowly add 0.5mL 20mg / mL rHrpZ into 5mL 50mg / mL PLGA solution dropwise (control the drop within 0.5 minutes), and at the same time emulsify with an ultrasonic breaker (50W treatment for 1 minute) to form a water-in-oil (W / O) emulsion, The water-in-oil emulsion was immediately poured into 50 mL of 2% polyvinyl alcohol (poly vinyl alcohol, PVA, molecular weight 70,000) aqueous solution. Ultrasonic emulsification (100W treatment for 2 minutes) forms a W / O / W emulsion. Pour the W / O / W emulsion into a distillation flask, distill under reduced pressure at 40°C for 2 hours to remove dichloromethane, solidify PLGA to form nanoparticles, and form rHrpZ-PLGA nanocomposite (rHrpZ-PLGA NP).

[00...

Embodiment 3

[0071] Example 3: Determination of Harpin Protein Nanocomplex Particle Diameter

[0072] The prepared rHrpZ-PLGA nanoparticles were resuspended in 10ml of water, 3ml was added to a quartz cuvette, and the sample temperature was set at 20°C. The particle diameter of the synthesized rHrpZ-PLGA nanocomposite was measured with a dynamic light scattering particle diameter detector. The test results show that 72.5% of the prepared nanoparticles have a diameter smaller than 225nm, and the average particle diameter is 190.5nm. The diameter distribution of nanoparticles is shown in the attached image 3 shown.

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Abstract

The invention discloses a protein nano complex promoting plant growth and a preparation method and application thereof. A medical nano controlled-release system is adopted to couple a plant immunity activator Harpin protein with bioactivity with a carrier to prepare the nano complex. The Harpin protein with the bioactivity is obtained through separation and purification. Nanoparticles are prepared by PLGA, PVA and ultrasonic technology, and coat the Harpin protein to form the PLGA-protein nano complex carrying the Harpin protein. The protein nano complex prepared by the method comprises the following advantages that: (1) the bioavailability of the Harpin protein can be improved after the Harpin protein is coupled with the ultramicron carrier; (2) the Harpin protein is appropriately coated, and the protein can be protected; (3) the protein nano complex can promote a plant to absorb and utilize a protein medicament, the effect is strengthened or prolonged and obvious biological effects are generated; (4) the protein nano complex has simple and safe use; and (5) the protein nano complex has disease-resistant broad spectrum and does not pollute environment. The protein nano complex can be used for preventing and controlling plant diseases, promoting the growth of plants and improving crop yield.

Description

technical field [0001] The invention relates to the fields of medicine and plant disease prevention and control, and more specifically, the invention relates to a preparation method and application of a protein nanocomposite for promoting plant growth. A drug nanometer controlled release system is used to couple the biologically active plant immune activator Harpin protein with a carrier to prepare nanoparticles. The preparation can protect the Harpin protein, promote the absorption and utilization of the medicine by plants, enhance or prolong the efficacy of the medicine, increase the bioavailability of the medicine, and produce obvious biological effects. This is a high-efficiency, broad-spectrum and safe new biopesticide for plant disease control. Background technique [0002] Plant diseases and insect pests have long been the main agricultural disasters that threaten my country's agricultural production and national economic development. The annual loss of crops due to...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N47/44A01N25/04A01P1/00A01P3/00
Inventor 徐进平孟小林王健梁焯
Owner WUHAN UNIV
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