Belonging to the technical field of
plant tissue culture, the invention provides a method for introducing
nitrogen-fixing
bacteria into sugarcane
tissue culture seedlings. The method consists of: under aseptic operation conditions,
cutting the stem tips of sugarcane, and conducting
callus induction,
seedling differentiation culture, rooting induction as well as strong
seedling culture in order; making use of a labeled strain of
Klebsiella to prepare a bacterial suspension, and carrying out bacterium
inoculation treatment on sugarcane
tissue culture seedlings; determining the
colonization dynamics of the labeled strain of
Klebsiella in the sugarcane tissue culture seedlings; and detecting
colonization of the labeled strain of
Klebsiella in sugarcane tissues by a
fluorescence microscope. The method for introducing
nitrogen-fixing
bacteria into the sugarcane tissue culture seedlings adopts the
fluorescence microscope to observe infection and
colonization of the Klebsiella labeled strain, investigates the
population dynamics of the Klebsiella labeled strain by an
antibiotic resistance labeling method and a plate isolating counting method, and provides references for popularization and application of health seedlings with
nitrogen-fixing
bacteria and for the interaction study of nitrogen-fixing bacteria and sugarcane, thus having high promotion and the application value.