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Method for screening citrus HLB (Huanglongbing) control agents through grafting

A technology of citrus huanglongbing and huanglongbing bacteria, which is applied in grafting, botany equipment and methods, applications, etc., can solve the problems of the influence of screening results, lack of evaluation system, complicated screening process, etc., and achieve reliable results, accurate evaluation system, and technical mature effect

Inactive Publication Date: 2016-08-10
POMOLOGY RES INST FUJIAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these systems are not carried out on citrus plants, but use intermediate hosts for screening of control agents, which may have a certain impact on the screening results
All in all, the current process of screening pesticides for controlling Huanglongbing on citrus is complicated, and there is a lack of a reliable and effective evaluation system

Method used

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  • Method for screening citrus HLB (Huanglongbing) control agents through grafting
  • Method for screening citrus HLB (Huanglongbing) control agents through grafting
  • Method for screening citrus HLB (Huanglongbing) control agents through grafting

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Embodiment 1: Grafting of diseased scion after medicament treatment

[0027] ① Coordinating and arranging the test, the grafting behavior is generally selected when the ambient temperature is 20 ℃;

[0028] ②Choose 2-year-old Fructus aurantii seedlings, the main branch diameter is about 15mm, cut off with a pruning knife at about 10 cm from the soil, level the section, and keep a 5° slope;

[0029] ③Take out the scion, choose a plump bud, cut it into a slope 2cm below the back of the bud, the slope and the longitudinal axis form an angle of about 30°, cut off the cortex at the tip, and separate the cortex from the xylem until the bud is cut off;

[0030] ④ On the cross-section of the rootstock, on the premise of not harming the xylem, use a grafting knife to cut vertically on one side of the rootstock from top to bottom;

[0031] ⑤Insert the cut scion into the rootstock with the oblique side facing down, without leaving a wounded surface, and tie the rootstock and the ...

Embodiment 2

[0033] Example 2: Detection of Citrus Huanglongbing Healthy Plants and Infected Scions

[0034] ①Extraction of total DNA from citrus leaves (CTAB method)

[0035] Cut 0.5 g of the leaf midrib, place it in a mortar, and grind it into a fine powder with liquid nitrogen; transfer the fine powder to a 2 mL centrifuge tube, add 900 μL 2% CTAB, 10 μL β-mercaptoethanol, 90 μL 10% SDS, 2 μL proteinase K; 65°C, water bath for 1 h, during which time gently shake once every 10 min; cool to room temperature, 12000 rpm centrifugation for 10 min; take the supernatant, add an equal volume of atmosphere / chloroform / isoamyl alcohol (25: 24:1), gently inverted, let stand for 5 minutes, and centrifuge at 12,000 rpm for 10 minutes; take the supernatant, add an equal volume of chloroform / isoamyl alcohol (24:1), gently turn it upside down, let stand for 5 minutes, and centrifuge at 12,000 rpm for 10 minutes; Take the supernatant, add an equal volume of isopropanol (or 2.5 times the volume of absolu...

Embodiment 3

[0038] Embodiment 3: the growth situation of scion and the survival rate of grafting after different medicament treatment

[0039] Such as figure 1 As shown, the scion began to germinate about 15 days after grafting, and germinated completely after 30 days. At this time, the grafting wrapping film should be lifted to guide the buds out of the bag; 45 days after grafting, the buds have grown into branches with a length of about 4 cm. At this time, all the grafting wrapping films were carefully removed; 60 days after grafting, the lower leaves of the branches began to mature, and after another month, samples could be taken after the leaves were fully mature to detect the content of pathogens. From figure 2 and image 3 From the results, it can be clearly seen that when the scion was treated with sodium dichloroisocyanurate (NaDCC) and tetracycline hydrochloride (TET), the overall graft survival rate was relatively high, but the graft survival rate declined with the increase o...

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Abstract

The invention belongs to the technical field of control of fruit tree diseases and particularly discloses a method for screening citrus HLB (Huanglongbing) control agents through grafting. According to the method for screening the citrus HLB (Huanglongbing) control agents, a citrus scion infected with the disease is taken as an object, the content change of Candidatus liberibacter asiaticus in the scion infected with the disease and treated with the agents is detected quantitatively through a real-time PCR (polymerase chain reaction), and the control effects of the treating agents are evaluated on the basis. The citrus scion infected with the disease is taken as a study object directly, the agent efficacy evaluation more conforms to practical situations, and influence caused by an interhost such as Diaphorina citri Kuwayama or catharanthus roseus used in screening of Candidatus liberibacter asiaticus control agents is eliminated. The problems that a screening process of the Candidatus liberibacter asiaticus control agents is complicated, a reliable and effective evaluation system is absent and the like at present are solved, and reliable technical and theoretical bases are provided for control of citrus HLB.

Description

technical field [0001] The invention relates to a method for screening agents for prevention and control of fruit tree diseases, in particular to a method for screening agents for prevention and control of citrus huanglongbing by grafting, and belongs to the technical field of disease prevention and control for fruit trees. Background technique [0002] Citrus Huanglongbing (HLB) is the most destructive disease in citrus production, and it is also a key quarantine target in major citrus producing areas. The pathogen of citrus huanglongbing in my country belongs to the Asian species (Candidatus Liberibacterasiaticus, Ca. L. asiaticus), and it is the most widely damaged huanglongbing fungus. In HLB research, the isolation and cultivation of pathogenic bacteria is a key link and also a bottleneck of current research work. Although Davis et al. (2008) and Sechler et al. (2009) obtained pathogenic bacteria through co-cultivation and pure culture respectively, none of the obtaine...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01G1/06A01G13/00
CPCA01G2/30A01G13/00
Inventor 王贤达范国成胡菡青林雄杰
Owner POMOLOGY RES INST FUJIAN ACAD OF AGRI SCI
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