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Inoculation method of Chinese wheat mosaic virus (CWMV)

A technology of wheat mosaic virus and wheat, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of low friction inoculation efficiency, slow research progress, cumbersome steps, etc., to improve inoculation rate, simplify inoculation Poison steps, simple operation effect

Active Publication Date: 2020-09-29
NINGBO UNIV
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  • Abstract
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AI Technical Summary

Problems solved by technology

At present, wheat is inoculated with CWMV virus by frictional inoculation. Due to the low efficiency of frictional inoculation, cumbersome steps and high cost, the research on the interaction between wheat and CWMV has slowed down.

Method used

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  • Inoculation method of Chinese wheat mosaic virus (CWMV)
  • Inoculation method of Chinese wheat mosaic virus (CWMV)
  • Inoculation method of Chinese wheat mosaic virus (CWMV)

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preparation example Construction

[0020] In the present invention, the CWMV RNA1, CWMV RNA2 and P19 silencing suppressors in the CWMV infiltration fluid are preferably present in the form of transformed Agrobacterium culture fluid. In the present invention, the nucleotide sequences of the CWMV RNA1, CWMV RNA2 and P19 are respectively shown in SEQ ID No.1-3. In the present invention, the preparation method of the transformed Agrobacterium culture solution comprises the following steps: respectively transforming CWMV RNA1, CWMV RNA2 and P19 silencing suppressor into GV3101 Agrobacterium by electroporation transformation, and using 50 μg / ml card Napenicillin + 50 μg / ml gentamycin + 50 μg / ml rifampicin three kinds of antibiotics YEP culture medium were shaken at 28 ℃ for 14 ~ 16 hours to obtain CWMV RNA1-transformed Agrobacterium culture fluid, CWMV RNA2 transformed Agrobacterium culture fluid and P19 silencing suppressor-transformed Agrobacterium culture fluid. In the present invention, the CWMV RNA1-transformed...

Embodiment 1

[0026] 1) Soak wheat seeds in tap water at 25°C for 12 hours overnight;

[0027] 2) Place the seeds with white buds on absorbent paper for moisturizing and culturing for 3 days, and obtain wheat seedlings with a total length of stem and leaf of 4-8 cm;

[0028]3) Transform CWMV RNA1, CWMV RNA2, and P19 silencing suppressors into GV3101 Agrobacterium by electric shock transformation method, and use 50 μg / ml kanamycin + 50 μg / ml gentamicin + 50 μg / ml rifampin The YEP medium of the three antibiotics was shaken at 28°C for 15 hours to obtain the CWMV RNA1-transformed Agrobacterium culture fluid, the CWMV RNA2-transformed Agrobacterium culture fluid and the P19 silencing suppressor-transformed Agrobacterium culture fluid.

[0029] 4) MES buffer (10mM MES; 10mM MgCL 2 ; 0.2mM AS) to dilute the collected thalline after centrifugation to prepare CWMV RNA1:CWMV RNA2:P19 OD 600 = 1:1:1 CWMV infiltration solution; OD of CWMV RNA1-transformed Agrobacterium cells, CWMV RNA2-transformed A...

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Abstract

The invention provides an inoculation method of Chinese wheat mosaic virus (CWMV), and belongs to the technical field of plant virus inoculation. The inoculation method comprises the following steps:placing wheat seedlings with stem and leaf total length of 4-8 cm in a CWMV infiltration liquid, performing vacuum pumping treatment for 25-35 min under standard atmospheric pressure, and then transplanting the wheat seedlings into soil for culture, wherein the CWMV infiltration liquid comprises CWMV RNA1, CWMV RNA2 and P19 silencing suppressors. By the inoculation method provided by the invention, the inoculation rate of the Chinese wheat mosaic virus (CWMV) on wheat can be significantly improved, the inoculation steps are simple, and rapid batch inoculation can be realized.

Description

technical field [0001] The invention belongs to the technical field of plant virus inoculation, and in particular relates to an inoculation method of Chinese wheat mosaic virus CWMV. Background technique [0002] Chinese wheat mosaic virus (CWMV) is a soil-borne virus transmitted by Polymyxa graminearum identified on winter wheat in Shandong Province in the 1990s (Chen J. Annals of Applied Biology, 1993, 123 (1):55-61) In recent years, the disease area infected by CWMV has been continuously expanding, resulting in a 10%-30% or even 70% reduction in wheat yield. In the winter of 2018, due to abundant rainfall and long duration of low temperature, the incidence of wheat mosaic virus disease in China was particularly serious. So far, no effective and targeted prevention and control measures for soil-borne virus diseases have been developed. The only effective control method in production is to breed disease-resistant varieties, but resistant germplasm resources are relatively ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01G7/00
CPCA01G7/00Y02P60/14
Inventor 钟凯丽程晔张帆刘婷婷刘芃羊健陈剑平
Owner NINGBO UNIV
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