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Rapid inoculation and identification method for citrus bacterial canker disease

A technology of citrus canker and its identification method, which is applied in the field of rapid inoculation and identification of citrus canker, can solve the problems of insufficient leaf nutrient and water supply, complex management of water, fertilizer and pests, and large space for cultivation environment, and achieve rapid and efficient screening, The identification results are reliable and the effect of reducing the influence of environmental factors

Active Publication Date: 2020-07-31
HUNAN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Both in vitro and in vivo identification methods have different defects. For example, in vitro identification means that the leaves are separated from the mother tree, the supply of nutrients and water to the leaves is insufficient, the disease is unstable, and the leaves are easy to dry and die; although the in vivo identification is relatively stable, the operation Inconvenient, the required cultivation environment takes up a lot of space, the demand for inoculated seedlings is also large, and the management of water, fertilizer, diseases and insect pests is more complicated

Method used

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  • Rapid inoculation and identification method for citrus bacterial canker disease
  • Rapid inoculation and identification method for citrus bacterial canker disease
  • Rapid inoculation and identification method for citrus bacterial canker disease

Examples

Experimental program
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Embodiment 1

[0027] The operation steps of this embodiment are as follows:

[0028] (1) Sample collection: Pick 3 tender leaves that have reached the maximum leaf area but not completely turned green from the shoots of the plant, put them in a ziplock bag and mark the samples; each identification uses the disease-resistant variety Citron C -05 and the susceptible variety Bingtang Orange as control;

[0029] (2) Cultivation of pathogenic bacteria: pick a single colony of ulcer disease from the preserved solid medium plate, add 20 mL liquid LB medium, shake culture in a constant temperature shaker at 28°C, 200 r / min for 20 h, use sterile water Dilute the pathogen suspension to 10 3 , 10 4 , 10 5 , 10 6 cfu / ml;

[0030] (3) Identification of ulcer inoculation of the sample: After the sample is taken back to the laboratory, wipe the front and back of the leaf with alcohol cotton, turn the leaf over until the back of the leaf is facing upwards. After the alcohol evaporates, use a 1 mL syringe needle ...

Embodiment 2

[0035] (1) Sample collection: Pick 3 tender leaves that have reached the maximum leaf area but not completely turned green from the plant shoots, put them in a ziplock bag and mark the samples; each identification uses the disease-resistant variety Citron C -05 and the susceptible variety Bingtang Orange as control;

[0036] (2) Cultivation of pathogenic bacteria: pick a single colony of ulcer disease from the preserved solid medium plate, add 20 mL of liquid LB medium, shake culture in a constant temperature shaker at 28°C, 200 r / min for 20 h, use sterile water Dilute the pathogen suspension to 10 3 , 10 4 , 10 5 , 10 6 cfu / ml;

[0037] (3) Identification of ulcer inoculation of the sample: After the sample is taken back to the laboratory, wipe the front and back of the leaf with alcohol cotton, turn the leaf over until the back of the leaf is facing upwards. Puncture a small hole (do not pierce the leaf), and then inject the resuspended bacteria liquid into the mesophyll tissue ...

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Abstract

The invention provides a rapid inoculation and identification method for a citrus bacterial canker disease. The method comprises the following steps of (1) sample leaf collection; (2) pathogen cultureof a citrus bacterial canker disease; (3) identification of the canker disease inoculated into sample leaves; (4) rooting culture of the sample leaves; and (5) culture management of the sample leaves. Aiming at the disadvantages and shortcomings of existing resistance identification of the citrus bacterial canker disease, culture and identification modes are innovated through in vitro culture ofthe leaves inoculated with the canker disease for rooting, supplementing nutrients and water and increasing growth and observation periods. The rapid inoculation and identification method is simple inoperation, efficient and rapid in inoculation and identification, reliable in structure and more intuitive.

Description

Technical field [0001] The invention relates to a method for detecting citrus canker, in particular to a method for rapid inoculation and identification of citrus canker. Background technique [0002] Citrus is the largest fruit in the world. Citrus canker is a worldwide quarantine disease that seriously hinders the development of the citrus industry, and it is difficult to prevent and control. [0003] Citrus canker is caused by Xanthomonas citri subsp. citri. Its growth temperature is 5~36℃. The most suitable temperature for canker bacteria growth is 25~30℃, and the lethal temperature is 55~65. ℃. Citrus bacterial canker disease (Citrus bacterial canker disease) has disadvantages that are difficult to prevent and cannot be eradicated. The disease has strong transmission ability. Once it occurs in the new area, it requires a lot of manpower and material resources to prevent and control it, causing great economic losses. Breeding disease-resistant varieties is the only effective ...

Claims

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Application Information

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IPC IPC(8): A01H4/00C12N1/20C12Q1/18C12R1/64
CPCA01H4/008C12N1/20C12Q1/18
Inventor 马先锋朱志媚邓子牛戴素明盛玲
Owner HUNAN AGRICULTURAL UNIV
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