A method for rapidly transfecting citrus huanglongbing bacteria into non-toxic citrus psyllids

By employing standardized adhesive plating, insect fixation in different stages, and microinjection techniques, combined with constant temperature and humidity culture, the problem of inconsistent transfection of Huanglongbing pathogen in citrus was solved, achieving safe and efficient pathogen transfection and sample homogeneity.

CN122427809APending Publication Date: 2026-07-21NANCHANG UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
NANCHANG UNIV
Filing Date
2026-04-19
Publication Date
2026-07-21

AI Technical Summary

Technical Problem

Existing technologies cannot efficiently and safely transfer citrus Huanglongbing pathogens to non-toxic citrus psyllids, and there are risks of disease spread and inconsistencies in samples.

Method used

A stable and controllable transfection method was established by using adhesive plate disinfection and viscosity reduction treatment, insect fixation pretreatment, quantitative microinjection, and constant temperature and humidity culture. The bacterial titer, injection volume, and culture conditions were clearly defined to avoid mechanical damage and the spread of pathogens.

Benefits of technology

This method enables rapid and controllable pathogen transfection, yields uniformly sized infected insects, reduces the risk of disease spread, and improves the uniformity of experimental samples and the success rate of transfection.

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Abstract

The application discloses a method for quickly transfecting citrus Huanglongbing bacteria into non-toxic citrus psylla, relates to the technical field of entomology and microbiology, and specifically comprises the following steps: preparing a sticking plate for fixing the psylla, and completing disinfection and viscosity reduction treatment; selecting non-toxic citrus psylla, fixing the psylla with the abdomen upward after completing pretreatment, taking citrus Huanglongbing bacteria liquid with a set titer, quantitatively injecting the bacteria liquid into the fixed psylla through a microscopic device, transferring the injected psylla to host plant tissues, and placing and culturing the psylla in a constant-temperature and constant-humidity environment to complete the bacteria transfection treatment; the method is characterized in that the standardization of the sticking plate preparation and the microinjection process of the separated psylla, the standardization of the disinfection, viscosity reduction and injection operation, the avoidance of the damage of the psylla and the guarantee of the survival rate, the accurate regulation and control of the parameters of the bacteria liquid and the culture conditions, the avoidance of the blockage of the needle, the efficient preparation of the homogenized bacteria-carrying psylla without the need of the diseased plants, the reduction of the disease spreading risk and the improvement of the experimental reproducibility.
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