Method for introducing nitrogen-fixing bacteria into sugarcane tissue culture seedlings

Abstract

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.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for introducing nitrogen-fixing bacteria into sugarcane tissue culture seedlings. Background technique [0002] Healthy sugarcane seedlings have the advantages of fast growth, strong tillering ability, high stem formation rate, high yield, and less seed usage. It is one of the important measures to ensure the sustainable development of the sugarcane industry. The country also attaches great importance to the promotion of healthy sugarcane seedlings. In 2011, the Ministry of Agriculture launched the demonstration and promotion of sugarcane healthy seedlings in 4 provinces (regions) of Guangxi, Yunnan, Guangdong and Hainan. In the world's major sugarcane producing countries, such as Brazil, Cuba, the United States, Australia and other countries, 90% of the sugarcane areas have realized the healthy planting of sugarcane production seeds, so that th...

AI Technical Summary

Problems solved by technology

[0006] The present invention provides a method for introducing nitrogen-fixing bacteria into sugarcane tissue culture seedlings, aiming to solve the problems of introducing solid nitrogen into sugarcane tissue culture seedlings in the prior art, which have complicated introduction process, high cost and insignificant effect

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