The invention discloses a method for separating and identifying
pathogenic bacteria of insects, relating to the field of research for bacterial diseases of insects. The method comprises the followingsteps: S1, disinfecting the body surfaces of the insects carrying
pathogenic bacteria, extracting
hemolymph, carrying out diluting till the suitable concentration is achieved,
coating an LB plate withthe
diluent, carrying out culturing, picking out single colonies, thus obtaining
pathogenic bacteria, carrying out pure culturing, transferring to liquid LB culture medium, and observing the morphological features of isolated strains under an
oil immersion lens; S2, treating the
thallus obtained by culturing in S1 by using
lysozyme and SDS in sequence, and after the
thallus is crushed, purifyingthe total
DNA of the isolated strains through
phenol extraction; and S3, amplifying the 16S rDNA with overall length of the isolated strains, after sequencing, carrying out clustering analysis, and determining the species of the isolated strain in combination with the morphological features of an isolated strain. The method for separating and identifying pathogenic
bacteria of insects is universalfor
gram-positive
bacteria and
gram-negative
bacteria, and is suitable for separating and identifying pathogenic bacteria of the insects including bees, cotton bollworms, inchworms,
Bombyx mori and the like, and the screened 16S rDNA amplification and sequencing primers are good in universality and high in stability.