Preparation method and application of catharanthus roseus high-susceptibility liberobacter asiaticum model

A technology for citrus huanglongbing and periwinkle, applied in the biological field, can solve the problems of slow grafting, cumbersome, unfavorable drug development and the like

Active Publication Date: 2021-10-01
GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Citrus huanglongbing is a common bacterial disease of citrus, which has the characteristics that pathogenic bacteria cannot be cultured in vitro, have a long incubation period in citrus plants, and show symptoms slowly. Therefore, citrus plants themselves are not suitable for research on Huanglongbing
It is found in the prior art that periwinkle is the natural host of citrus huanglongbing bacteria, and the pathogenic bacteria can reproduce rapidly in it. At present, there are also reports about using periwinkle to prepare citrus huanglongbing model plants. For example, the application number is: 201310691324.X , the name of the invention is "Method for Screening Anti-phloem Bacteria Drugs Using Periwinkle Flowers": This method is also a method for quickly screening drugs using periwinkle flowers as a test material, but the method is to connect the haustoria of dodder to the vascular bundle of the host plant , the grafting speed is slow and the success rate of inoculation is not high; through a large number of literature reviews, it is found that in the prior art, most of them are inoculated by using periwinkle branches, and the infection rate can be guaranteed through branch inoculation, but most of them need to be bundled. To complete the entire grafting process, the grafting efficiency is slow and cumbersome
[0003] In the process of drug screening for citrus Huanglongbing, a large number of model plants are needed. If the grafting method is too cumbersome and complicated, it is not conducive to the development of highly susceptible citrus Huanglongbing model plants. Establishment is not conducive to large-scale intensive, clustered and large-scale screening, and the progress of drug screening is slow, which is not conducive to the development of drugs. Therefore, it is necessary to improve the inoculation method and invent an efficient and rapid The inoculation method can not only improve the grafting efficiency of highly susceptible plants, but also further improve the screening efficiency of drugs against citrus greening disease. Conduct tests to find out the best way to use ampicillin as the positive control drug, the time of drug use, and the concentration of drug use, so as to quickly screen and obtain small molecule drugs with significant disease resistance, and provide a theoretical basis for the prevention and treatment of citrus Huanglongbing. Exploring new ways to prevent and treat with small molecule drugs

Method used

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  • Preparation method and application of catharanthus roseus high-susceptibility liberobacter asiaticum model
  • Preparation method and application of catharanthus roseus high-susceptibility liberobacter asiaticum model
  • Preparation method and application of catharanthus roseus high-susceptibility liberobacter asiaticum model

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] This embodiment is a method for preparing a high-sensitivity Huanglongbing periwinkle model, the method comprising the following steps:

[0050] (1) Cultivate periwinkle plants: select periwinkle seeds to sterilize, accelerate germination, and germinate and cultivate. During cultivation, select healthy periwinkle plants with good growth and strong branches as stock, remove the terminal buds of stock, and open on stock;

[0051] (2) select the blade or terminal bud of the periwinkle diseased plant that has been infected with Huanglongbing and show yellowing and mottled disease as the scion;

[0052] (3) After inserting the scion leaves into the rootstock incision and aligning them tightly, select a grafting clip suitable for the size and thickness of the branches to clamp from the side where the scion is inserted, so as to avoid crushing the plant or extruding the scion due to too small a grafting clip, or The grafting clamp is too large so that the contact between the s...

Embodiment 2

[0065] This embodiment is a further study carried out at Example 1, mainly for the impact of different grafting materials and grafting methods on the periwinkle infection rate:

[0066] The specific scheme is as follows: in preferred embodiment 1, the periwinkle plants with a disease-susceptibility level III are used as materials for grafting, and the buds and leaves of the periwinkle plants with a disease-susceptible level III are selected as scions, and grafted by membrane grafting and clips way to accomplish the grafting:

[0067] ① Film grafting methods such as figure 2 Shown: Use a scalpel to cross-cut the terminal bud above the first pair of complete leaves on the top of a healthy periwinkle branch and remove it. On the cut surface, cut a joint of about one centimeter from top to bottom at the position of the phloem. The petiole of the scion is cut into a wedge shape at one centimeter and inserted into the cut of the rootstock in time to align and stick tightly, and th...

Embodiment 3

[0081] Through the research of embodiment 1 and embodiment 2, the applicant finds: adopting susceptible leaf periwinkle to be scion grafting periwinkle plant can effectively improve the establishment efficiency of periwinkle high sensitivity Huanglongbing model plant, therefore, the applicant in embodiment 1 On the basis of this study, optimization experiments were carried out on the establishment of periwinkle high-sensitivity Huanglongbing model plants, the details are as follows:

[0082] When the healthy periwinkle plant used as a rootstock grows to 5-9 pairs of true leaves, the top is pinched, and then the leaves of the periwinkle plant with a disease-susceptible level III are used as the scion. When the leaves are taken, the yellowing of the leaves is calculated by the degree of yellowing Area ratio, the ratio of leaf yellowing area is divided into four grades:

[0083] Level 1: 1-25% yellowing area ratio; (such as Figure 4 )

[0084] Level 2: 26-50% yellowing area ra...

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Abstract

The invention relates to the technical field of biology, in particular to a preparation method and application of a catharanthus roseus high-susceptibility liberobacter asiaticum model. Liberobacter asiaticum pathogenic bacteria are inoculated into healthy catharanthus roseus plants in a leaf grafting mode, when the method is used, the catharanthus roseus plants can be rapidly infected with the liberobacter asiaticum by controlling the susceptible level of inoculated leaves and the growth state of stocks and through later-stage moisturizing management, and a reliable and effective method is provided for obtaining the catharanthus roseus high-susceptibility liberobacter asiaticum model in batches. Through comparison, compared with a traditional film grafting method, the efficiency of the method is improved by three times, and the plant susceptibility rate is equivalent to that of an existing grafting method; meanwhile, ampicillin is used as a positive control on the basis of the model, and the model can be used for screening the medicine for inhibiting the liberobacter asiaticum; and meanwhile, the optimal application condition of the ampicillin is found out.

Description

【Technical field】 [0001] The invention relates to the field of biotechnology, in particular to a preparation method and application of a periwinkle highly susceptible citrus huanglongbing model. 【Background technique】 [0002] Citrus huanglongbing is a common bacterial disease of citrus. It has the characteristics that the pathogenic bacteria cannot be cultured in vitro, has a long incubation period in citrus plants, and slow symptoms. Therefore, citrus plants themselves are not suitable for research on huanglongbing. It is found in the prior art that periwinkle is the natural host of citrus huanglongbing bacteria, and the pathogenic bacteria can reproduce rapidly in it. At present, there are also reports about using periwinkle to prepare citrus huanglongbing model plants. For example, the application number is: 201310691324.X , the name of the invention is "Method for Screening Anti-phloem Bacteria Drugs Using Periwinkle Flowers": This method is also a method for quickly sc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01G7/06A01G2/30A01G13/00A01G22/60
CPCA01G7/06A01G2/30A01G13/00A01G22/60Y02A40/25
Inventor 张金莲温云英宋娟陈廷速刘金华钟泽橙李栋张瑞峰张师音尚鹏祥李冬萍汪茜程通胡柳罗文新葛胜祥夏宁邵
Owner GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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