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Plant immunity activator protein PsPII1 and applications thereof

An immune activation and plant technology, applied in the fields of application, plant peptides, plant growth regulators, etc., can solve the problems of easy loss of crop resistance, lack of high-efficiency and low-toxic fungicides, and easy resistance of bacteria

Active Publication Date: 2020-05-19
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Phytophthora phytophthora is similar in shape to fungi, but it belongs to the Oomycota of the kingdom Antleria in classification, and many fungicides are ineffective against Phytophthora (Tyler, 2007)
Moreover, at present, there are problems in the prevention and control of

Method used

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  • Plant immunity activator protein PsPII1 and applications thereof
  • Plant immunity activator protein PsPII1 and applications thereof
  • Plant immunity activator protein PsPII1 and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Determination and screening of candidate proteins in Phytophthora soybean culture fluid

[0028] Phytophthora sojae Kaufmann et Gerdemann used in this example was purchased from ATCC (American Type Culture Collection, www.atcc.org), and the strain number is

[0029] Inoculate the Phytophthora sojae purchased above on a V8 medium plate, culture at 25°C for 7 days, pick the edge of the colony and inoculate it in 200mL PDB liquid medium (the container is a 500mL Erlenmeyer flask), shake at 25°C and 180r / min. Bed culture was carried out for 5 days to obtain a culture solution of Phytophthora sojae.

[0030] The preparation method of the above-mentioned V8 medium plate: add 1g CaCO 3 Add it to 100mL V8 juice, mix and stir on a magnetic stirrer for 10min, 3000rpm, centrifuge at room temperature for 5-7min, take the supernatant and deionized water and dilute it at a ratio of 1:9 to make 10% V8 culture solution, add Agar powder 15g, boiled for 10 minutes to dissolve, subpac...

Embodiment 2

[0038] Cloning of the gene encoding plant immune activation protein PsPII1

[0039] Step 1): total RNA extraction

[0040] The hyphae of Phytophthora soybean in liquid culture was used as the material, and the total RNA was extracted using the RNA extraction kit of Omega Company, and the specific operation was carried out according to the instructions; the content and quality of the obtained total RNA were detected by a spectrometer.

[0041] Step 2): Reverse transcription to generate first strand

[0042]Take 0.7 μg of the total RNA obtained in the above step 1) as a template, and use Takara’s PrimeScript Reverse Transcriptase Kit for cDNA synthesis (see the kit manual for specific operations, and set the volume to 20 μL for reaction). The obtained reverse transcription product cDNA was used as a template for subsequent gene cloning PCR.

[0043] Step 3): RT-PCR amplification of the full length of the PsPII1 coding gene

[0044] The inventors designed the following PCR amp...

Embodiment 3

[0053] Transient expression of gene encoding PsPII1 in tobacco leaves

[0054] The pGR107::PsPII1-3HA plasmid obtained in Example 3 (proven correct) was transformed into Agrobacterium GV3101 by electric shock, and coated with LB (containing kanamycin 50 μg / mL, rifampicin 50 μg / mL) plate, 28 After culturing at °C for 48 hours, positive clones were picked and verified by PCR to obtain the correct clone (ie, a single colony of Agrobacterium GV3101 transfected with the pGR107::PsPII1-3HA plasmid), hereinafter referred to as the clone of Example 3.

[0055] In addition, a control clone, a single colony of Agrobacterium GV3101 transfected with the pGR107::GFP-3HA plasmid, was obtained.

[0056] The implementation 3 clones and the control clone were respectively scaled up and cultured. Specific operations: respectively inoculated into 2 mL of LB liquid medium (containing kanamycin 50 μg / mL, rifampicin 50 μg / mL) on a constant temperature shaker at 28°C, 200rpm Culture overnight to OD...

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Abstract

The invention relates to plant immunity activator protein PsPII1 and applications thereof, and belongs to the technical field of biological control of crops. The novel plant immunity activator protein, named as PsPII1, is discovered in the culture solutions of phytophthora sojae; and the plant immunity activator protein PsPII1 has strong activity, can induce the resistance and defense response ofplants, can be applied to the disease control of crops, and can be used for the disease control of soybeans, tomatoes, strawberries and tobacco, especially for the prevention of soybean root rot and tobacco mosaic diseases.

Description

technical field [0001] The invention relates to a plant immune activation protein PsPII1 and its application, belonging to the technical field of crop biological control. Background technique [0002] Phytophthora is an important threat to agricultural production. More than 180 species of Phytophthora have been discovered and identified, which can harm almost all dicotyledonous plants, and the diseases caused are called "plant diseases". For example, Phytophthora sojae, also known as P. sojae) can cause soybean root rot, which has a short incubation period and rapid onset, and is a devastating disease in soybean production. According to statistics, Phytophthora causes economic losses of more than 16 billion yuan in my country's potatoes, soybeans, vegetables and other crops every year. [0003] Although Phytophthora phytophthora is similar to fungi in morphology, it belongs to Oomycota of the kingdom Pleurophyte in classification, and many fungicides are ineffective agains...

Claims

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Application Information

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IPC IPC(8): C07K14/415A01N37/46A01N43/36A01P1/00A01P3/00
CPCC07K14/415A01N37/46A01N43/36
Inventor 杨波王源超王雨音郑文跃柳泽汉
Owner NANJING AGRICULTURAL UNIVERSITY
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