A method for immobilizing pine xylophilus worms for microinjection

Abstract

The invention discloses a worm body fixation method for microinjection of pine xylophilus, belongs to the field of biotechnology, and specifically relates to the following steps: using SDS-DTT solution to soften the cuticle of the nematode; using pronase E solution to digest the cuticle of the nematode; Prepare a fixed pad with 3% agarose as the main raw material; pick the worm body from the water to the fixed pad with an eyelash needle; quickly press down the movable worm body with an eyelash needle under a dissecting microscope; when the water film of the whole body of the worm body is absorbed by the fixed pad At the same time, cover with injection oil. The invention successfully immobilizes the pine wood nematode, the fixation rate is more than 80%, and the survival rate of the pine wood nematode after injection is more than 80%.

Description

Technical field [0001] The present invention relates to a fixing method for microscopic pine lines, and specific to molecular biology. Background technique [0002] Pine Materials Bursaphelenchus Xylophilus is a parasitic nematode living in the pine. After the pine tree is infected by pine, it can be died in two months. Pine wood nematode adult insects is about 1 mm long, and the tail of female insect is close to the tapered, the end circle; the male tail is like a bird feet, bent to the abdomen. Pine material nematodes are achieved with the spread of pine trees by means of media insects such as Moqua, and triggers the prevalence of pine wood nematode disease. Pine Tree needles infected by the pinemar nematode, yellowish brown or red brown, and the whole plant died. [0003] As an important pathogen nematode, the basic biology research of pine-like nematodes is the frontier and hotspots of current forest pathology research. However, due to the lack of effective worm fixation tech...

AI Technical Summary

Problems solved by technology

Compared with Caenorhabditis elegans, the body wall of pine xylophilus is hard and thick, the composition of the cuticle on the body surface is complex and diverse, the protein content is high, and the adhesion to 2% agarose fixation pad is poor

In addition, compared with Caenorhabditis elegans, pine xylophilus is more mobile and can easily break free from the restraint of the fixed mat

Therefore, the immobilization of pine xylophilus cannot be effectively achieved by adopting the above-mentioned microinjection immobilization technology of C. elegans, and the immobilization rate is ≤10%.

Due to the poor immobilization effect, the pine wood nematode is still active during the injection process, and cannot be injected to the correct position accurately, causing greater mechanical damage to the worm body, and the survival rate of the pine wood nematode after injection is ≤20%.

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