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Virus stabbing inoculation method for cucurbitaceae crops

A technology of cucurbit crops and virus inoculation, applied in the biological field, can solve the problems of excessive damage to cells and plants, time-consuming and labor-intensive problems, and the inability to fix the amount of diseased sap.

Inactive Publication Date: 2016-02-24
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this method is that it is time-consuming and labor-intensive, and each inoculated plant must be sprayed, rubbed and cleaned back and forth; it has certain technical difficulties, and it will cause great damage to the plants. Excessive friction may cause cells or even plants to be damaged and die; There is a large error in the amount of seeds, and the amount of disease juice dipped in each inoculation cannot be fixed; the whole cycle is long, about 60-80 days from sowing to observation

Method used

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  • Virus stabbing inoculation method for cucurbitaceae crops
  • Virus stabbing inoculation method for cucurbitaceae crops
  • Virus stabbing inoculation method for cucurbitaceae crops

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Comparison of Morbidity Rate of Several Different Vaccination Methods

[0016] 1.1 Test materials and methods

[0017] The gourd seeds were sown in 32-hole hole trays and placed in the greenhouse until they germinated naturally and grew until two true leaves unfolded. Pick 1-2 diseased leaves infected with CGMMV, add a little quartz sand and an appropriate amount of phosphate buffer in a mortar, grind thoroughly, and prepare diseased juice for later use. The constructed invasive clones were activated and grown to OD600 of about 0.6 at 28°C at 200 rpm to collect the bacteria by centrifugation and resuspended with 1 / 2 MS, adding acetosyringone at a final concentration of 100 μmol / L for later use.

[0018] Friction inoculation: Sprinkle a little quartz sand on the leaves of the plant first, dip the diseased juice with your fingers and apply it back and forth gently.

[0019] Infiltration method: lightly puncture the back of the leaf with a syringe needle a few times, the...

Embodiment 2

[0026] Identification of Cucumber's Resistance to CGMMV by Acupuncture Inoculation

[0027] 1.1 Test materials and methods

[0028] Cucumber seeds (Table 1) of 15 different varieties were sown in 32-hole plug trays and placed in the greenhouse. After they germinated naturally and grew until the first true leaf unfolded, they were used for virus inoculation. Pick 1-2 diseased leaves infected with CGMMV, add a little quartz sand and an appropriate amount of phosphate buffer in a mortar, grind them thoroughly, and prepare diseased juice. Dip the diseased juice with an acupuncture needle and puncture the hypocotyl of the plant N times (N refers to 1-10 times), and dip the diseased juice once for each plant inoculated. The inoculated plants were placed in the greenhouse for normal management, and after 20 days of growth, the leaves of each plant were collected to detect the virus infection by ELISA.

[0029] 1.2 Test results

[0030] Through post-inoculation testing, it was foun...

Embodiment 3

[0035] Identification of Rootstock Pumpkin's Resistance to CGMMV by Acupuncture Inoculation

[0036] 1.1 Test materials and methods

[0037] Rootstock pumpkin seeds (Table 2) of 15 different varieties were sown in 32-hole plug trays and placed in a greenhouse. After they germinated naturally and grew until the first true leaf unfolded, they were used for virus inoculation. Pick 1-2 diseased leaves infected with CGMMV, add a little quartz sand and an appropriate amount of phosphate buffer in a mortar, grind them thoroughly, and prepare diseased juice. Dip the diseased juice with a crochet needle and prick N times at the hypocotyl of the plant (N refers to 1-10 times), and dip the diseased juice once for each inoculated plant. The inoculated plants were placed in the greenhouse for normal management, and after 20 days of growth, the leaves of each plant were collected to detect the virus infection by ELISA. Plants inoculated by rubbing at the same seedling age were used as the...

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Abstract

The invention provides an easy, convenient and efficient inoculation method for viruses and virus derivatives of cucurbitaceae crops, and belongs to the technical field of biology. The method is suitable for all viruses, capable of being subjected to mechanical inoculation, of cucurbitaceae crops and virus carriers obtained through infectious cloning and modifying of corresponding viruses, and whole operation is completed only in an aperture disc; the aim that plants are effectively infected by the viruses is achieved through stabbing inoculation. Compared with a traditional mechanical inoculation method, the method has the advantages that operation is easy and convenient, efficiency is high, the application range is wide, the inoculation amount is fixed, and an operating period is short; the method is a rapid, easy, convenient and efficient plant virus inoculation method and can be used for large-scale virus biological work, variety resistance screening and using, disease resisting variety breeding and producing of target bioactive substances with the viruses as carriers.

Description

technical field [0001] The invention relates to the application of a virus inoculation method in virus biology work, screening or breeding of disease-resistant varieties and production of biologically active substances with viruses as carriers, belonging to the field of biotechnology. Background technique [0002] In agricultural production, almost all kinds of food and vegetable crops have viral diseases. Among the vegetable crops, Solanaceae, Cucurbitaceae, Leguminosae, Cruciferae, etc. are more severely affected, and there are many types of viruses infected. Among them, viral diseases of cucurbit crops have occurred in almost all planting areas in the world. Among them, pumpkins are the most serious, followed by melons, cucumbers, watermelons, gourds and other crops are also infected to varying degrees. At present, there are as many as 48 kinds of viruses that harm Cucurbitaceae crops have been reported. The most common and serious ones are: Cucumber Mosaic Virus (CMV), ...

Claims

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Application Information

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IPC IPC(8): C12N15/82
CPCC12N15/8205C12N15/8207
Inventor 杨柳程兆榜任春梅陆芳缪倩
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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