A method for constructing fluorescently labeled rhizobia

Abstract

The invention discloses a method for using sterilized alfalfa leaves instead of sterile filter membranes as a three-parent hybrid carrier to improve the efficiency of fluorescent plasmid transfer conjugation and transfer of rhizobia fluorescently labeled strains, which can be used to efficiently construct rhizobia CFP cyan fluorescent protein genes To mark the bacterial strain, the method is as follows: take the young leaves of alfalfa, wash them with distilled water, place the leaves evenly in a single layer in a petri dish, and sterilize them; take out the leaves with sterile tweezers, and stick them on the surface of TY medium , and cover the Petri dish. In the process of three-parent hybridization, instead of the microporous membrane, the mixed growth culture of donor bacteria, auxiliary bacteria and recipient bacteria is carried out. Compared with the microporous filter membrane as the hybrid carrier, using alfalfa leaves as the hybrid carrier can increase the conjugative transfer rate of fluorescent plasmids by 8.4-11 times, and the difference reaches a significant level; because the raw materials used are renewable and easily obtained alfalfa plants The leaves, raw materials are cheap and easy to obtain, and the operation is simple and convenient, which is suitable for the construction of rhizobia fluorescently labeled strains.

Description

Technical field [0001] The present invention involves the field of crop vaccination, and specifically involves the construction method of a fluorescent alignment. Background technique [0002] Rhizoma vaccination agents are more commonly used in agricultural production. The yield of beancotic crops is also significantly improved due to the application of rhizoplasma inoculating agent, but the effectiveness, competitiveness and safety of the root tumor vaccination agent after the soil is applied to the soil.It also needs to be resolved.The development of root tumor labeling technology and its application in scientific research and actual production have gradually clarified the nitrogen fixation mechanism of rhinoma and its infection and colonization process in the root system of the bean family.One of the effective methods of competition and interaction with other microorganisms in the body.The current labeling effect is the best, and the most simple and easiest way to determine i...

AI Technical Summary

Problems solved by technology

However, due to the artificially prepared single-layer filter membrane structure, the nutrients in the medium can only be absorbed by the cells through the micropores, and the proliferation rate and conjugation transfer rate of the bacteria are limited.