Method of quantitatively evaluating cucumber green mottle mosaic virus resisting ability of Lagenaria leucantha

A quantitative evaluation, gourd technology, applied in the fields of botanical equipment and methods, measuring devices, instruments, etc., can solve the problems of hidden disease manifestations, complex CGMMV symptoms of gourd, inability to classify and describe, etc., to achieve system disease resistance, Accurate identification results and the effect of avoiding grading uncertainty

Inactive Publication Date: 2017-06-13
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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AI Technical Summary

Problems solved by technology

At present, gourd CGMMV resistance identification mainly uses the method of rubbing inoculation with virus-carrying fresh leaf juice. Although the operation is simple, it is difficult to ensure the singleness of the virus.
In addition, the symptoms of gourd CGMMV are complicated, and it is almost impossible to describe the traditional classification according to the plant performance. In addition, after CGMMV infection, sometimes hidden symptoms appear, making the accurate evaluation of the symptoms of CGMMV become the main method for identifying the resistance of gourd germplasm. Limiting factor
However, there is currently a lack of an objective and quantitative method to accurately evaluate the resistance of different gourd germplasm resources to CGMMV

Method used

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  • Method of quantitatively evaluating cucumber green mottle mosaic virus resisting ability of Lagenaria leucantha

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Analysis of the inoculation effect of different concentrations of invasive cloning bacteria:

[0022] (1) Plant material: Hangzhou long melon, CGMMV in the field.

[0023] (2) Inoculation identification: The seeds of Hangzhou long melon are sown in a nutrient bowl and grown in a glass greenhouse. The light time is 16 hours, and the temperature is controlled at 25-30°C during the day and 20-25°C at night. Inoculation working solution is OD 600 =1 Bacterial liquid diluted 10, 100, 1000, 10000 times, namely OD 600 =0.1, 0.01, 0.001, 0.0001 four concentrations. After the seedlings grow to the one-leaf-one-heart stage, they are inoculated by the cotyledon pressure contact method. Each individual plant produces 2 inoculation points, and 4 plants are inoculated at each concentration. At the same time, 4 plants are selected for inoculation buffer (Agrobacterium carrying empty vector) as a control (ck). Sampling was taken 15 days and 25 days after inoculation. The selection criteri...

Embodiment 2

[0028] Identification of germplasm resources of gourd gourd resistant to CGMMV

[0029] (1) Plant material: 173 germplasm resources of gourd gourd.

[0030] (2) Inoculation identification: All materials are uniformly sown in a nutrient bowl, with 6-10 plants for each material, arranged in 2 rows, and grown in a glass greenhouse. Use OD 600 =0.01 concentration of the bacterial liquid, after the cotyledons of each material are flattened to the one-leaf one-heart stage inoculation, press the back of the cotyledons with a 1ml syringe until the bacterial liquid penetrates into the leaves evenly. After inoculation, control the temperature at 25-30℃ during the day and 20- 25°C. All materials were sampled for the first time 15 days after inoculation. The selection criterion was the first flat leaf under the apical bud. Each material and each row of plants were sampled and mixed together to form 2 samples, which were grinded by liquid nitrogen and weighed. Add 15ml of the extraction solut...

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Abstract

The invention relates to the technical field of crop disease resistance and breeding, in particular to an inoculation identification and quantitative detection method of cucumber green mottle mosaic viruses of Lagenaria leucantha. A method of quantitatively evaluating the cucumber green mottle mosaic virus resisting ability of Lagenaria leucantha comprises the following steps: firstly, utilizing bacterial liquid carrying an infectious cloning vector to inoculate Lagenaria leucantha seedlings by a cotyledon pressing contact method, then detecting CGMMV absorption values in a Lagenaria leucantha plant within two time quantum after inoculation by utilizing an ELISA technology, and comprehensively evaluating the ability of resisting CGMMV of different Lagenaria leucantha germ plasm according to the content variation condition of the CGMMV. Therefore, the method replaces phenotypic identification based on disease symptoms and is simple to operate and liable to apply.

Description

Technical field [0001] The invention relates to the technical field of crop disease-resistant breeding, in particular to a method for inoculation identification and quantitative detection of gourd cucumber green mottle mosaic virus. Background technique [0002] Gourd[ Lagenaria siceraria (Molina) Standl.] Also known as gourd, long gourd, gourd, night flowering, gourd, etc., belong to the Cucurbitaceae ( Cucurbitaceae )Calabash ( Lagenaria ), an annual climbing herb. Gourd is an important melon vegetable crop in southern China in summer, and it is also the first choice for grafting watermelon and cucumber. Cucumber green mottle mosaic virus (CGMMV) is a newly introduced pest in my country in recent years. Due to the rapid spread of the virus and serious damage, it poses a great threat to gourd production. CGMMV belongs to the Tobacco Mosaic Virus (Tymoviridae) family ( Tobamovirus ), which mainly infects cucumbers, watermelons, melons, gourds, pumpkins and other cucurbitace...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/00G01N33/569
CPCA01H1/00G01N33/56983
Inventor 吴新义徐沛吴晓花郑红英燕飞汪宝根鲁忠富李国景
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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