Pepper cucumber mosaic virus seedling disease phase inoculation method and application thereof

A technology for cucumber mosaic virus and pepper, applied in the directions of application, horticultural methods, botanical equipment and methods, etc., can solve the problems of little use, difficult operation, large workload, etc., and achieves easy observation, reduced operation steps, and reduced Missing effect

Active Publication Date: 2018-05-18
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

The virus juice root soaking method refers to washing the soil at the root of the seedlings and immersing them in the inoculation s

Method used

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  • Pepper cucumber mosaic virus seedling disease phase inoculation method and application thereof
  • Pepper cucumber mosaic virus seedling disease phase inoculation method and application thereof
  • Pepper cucumber mosaic virus seedling disease phase inoculation method and application thereof

Examples

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Embodiment 1

[0027] (1) Take the purified strain of cucumber mosaic virus disease, propagate it on Nicotiana tabaccum CVSSamun NN, inoculate it by manual injection, and harvest the diseased leaves 12-15 days after inoculation. Add 5ml of 0.02Mol L with a Ph value of 7 per 1g of diseased leaves -1 Phosphate buffer homogenate in ice bath, filter with double gauze or 3500 rpm -1 Centrifuge for 15 minutes, and the filtrate or supernatant is the inoculum;

[0028] (2) Pepper seeds are rinsed 3-4 times with sterile water after being sterilized in 5% sodium hypochlorite for 20 minutes;

[0029] (3) Pepper seeds germinate in a petri dish under dark conditions at 25°C after disinfection, and sow them in an 8×8 nutrient pot for growth. Can be inoculated when fully expanded.

[0030] (4) Use a disposable syringe (1 mL) to draw 1 mL of the bacterial solution, remove the needle, and slowly inject the bacterial solution into the back of the pepper cotyledon, so that the two cotyledons are filled with...

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Abstract

The invention relates to a pepper cucumber mosaic virus seedling disease phase inoculation method and application thereof. The method is used for rapidly identifying and screening genetic resources with cucumber mosaic virus disease resistance in pepper resources of China. Samsun NN is manually inoculated with a purified cucumber mosaic virus source in an injected mode, diseased leaves are harvested 12-15 days after inoculation, phosphate buffer is added into the diseased leaves for ice-bath homogenization, filtering with double-layer gauze or centrifugalizing is carried out, and filtrate or supernatant is an inoculation solution. Inoculation can be carried out from the pepper cotyledon phase to the five-six-leaf phase, wherein the one-two-leaf phase is an optimal inoculation seedling age,1 mL of inoculation solution is sucked by a disposable syringe (1mL), a syringe needle is used for pricking two needle holes in each cotyledon, the syringe needle is removed, the inoculation solutionis slowly injected into the needle holes in the back face of each cotyledon, and the two cotyledons are full of the virus inoculation solution. Disease condition investigation is carried out 14 daysafter inoculation. The method has the advantages that the virus inoculation amount is easy to control, the inoculation solution is directly injected into mesophyll cells during inoculation, observation is easy, and missing inoculation is reduced.

Description

technical field [0001] The invention belongs to the field of plant protection, and relates to a seedling inoculation method for pepper cucumber mosaic virus disease and application thereof. Background technique [0002] The temperate zone is the main distribution area of ​​cucumber mosaic virus (CMV), and the tropical zone shows a trend of increasing year by year. So far, the host range of CMV exceeds 1200 species of plants in 100 families, and it is particularly harmful to a variety of vegetable crops and ornamental plants. CMV is mainly transmitted by a variety of aphids in a non-sustained manner, and some plants can be seed-borne. Due to the wide range of hosts, aphid transmission and low frequency of crop variety replacement, CMV can survive all year round, making it difficult to control. The detection rate of CMV in the main pepper producing areas in my country is very high, and the detection rates in Beijing, Shaanxi, Huguang, and Jiangsu and Zhejiang regions are all...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/18A01G7/06C12R1/94
CPCA01G7/06C12Q1/02C12Q1/18
Inventor 郭广君王述彬刘金兵潘宝贵刁卫平戈伟高长洲
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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