New method for obtaining ornamental arrowroot tissue culture sterile system

Abstract

Disclosed is a new method for obtaining ornamental arrowroot tissue culture germfree systems, which is an improved method for obtaining the germfree systems through the secondary transition planting of tissue culture female parent plant and the comprehensive sterilization of explants, by adding the sanitizers in the culture medium for co-cultivation; the plant tissue culture is made under sterile conditions; the process comprises the following steps: the female parent plant goes through secondary transition planting, namely, the female parent plant is cultivated in perlite or ceramic particles for 2-3 weeks and then removed into water for water culture to induce new leaves with less endophytes, and then the new leaves are taken as explants for tissue culture; the material goes through preliminary disinfection and low-temperature pretreatment; two disinfectant fungicides of different dosage are added into the induced culture medium to prepare the induced culture medium which can restrain bacteria; and the explants are treated through surface disinfection with an integrated approach. The method is unique, simple, practical and highly valued, which can obtain a large amount of aseptic systems in a short time, so as to save time, labor and expenses. The method enables the yield rate of the aseptic explants to reach 65-70%.

Description

technical field [0001] The invention belongs to the plant tissue culture technology in the field of biotechnology; in particular, it relates to a new method for obtaining an aseptic system of ornamental arrowroot tissue culture. Background technique: [0002] The factory production of plant tissue culture seedlings is developing rapidly in our country, and tissue culture technology is becoming more and more perfect. In market competition, how to reduce costs is the primary issue for improving economic benefits. The increase in the pollution rate in tissue culture will inevitably lead to the cost of tissue culture seedlings. Therefore, reducing the pollution rate in tissue culture is a technical link that cannot be ignored in factory production. [0003] There are many reasons for the pollution in the process of plant tissue culture, such as: incomplete disinfection of explants or pollution caused by endogenous bacteria; operational pollution and environmental pollution; poll...

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Problems solved by technology

[0004] During our experimental research, we found that it is very difficult to sterilize the explants (terminal buds or lateral buds) of Arrowroot plants during tissue culture. The key technical problem that solves; To make ornamental arrowroot tissue culture rapid propagation move toward industrialization from the laboratory, form commercialization, let more plant tissue culture seedlings go to the market, therefore must research a kind of effective, practical, fast external Implant disinfection method

[0005] Arrowroot buds are all located in the cultivation medium, and the underground part has relatively developed aerenchyma; in a large number of experiments we have conducted, through various methods and even some of them are particularly harsh disinfection conditions, but because there may be bacteria in the arrowroot tissue. Parasitic or symbiotic bacteria, most arrowroot tissue materials cannot be completely sterilized, resulting in the acquisition rate of sterile explants of arrowroot tissue materials being often lower than 5%

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