Tea tree virus-free tissue culture method

Abstract

The invention discloses a tea tree virus-free tissue culture method which comprises the following steps: (1) material selection: selecting the current-year branches which are robust without decay or diseases in spring; (2) surface treatment: cleaning and disinfecting the bud explant of the tea tree; (3) disinfecting; (4) cutting; (5) primary culture: inoculating to a culture medium I and culturing; (6) multiplication subculture: inoculating an aseptic seedling to a culture medium II, and performing multiplication culture; (7) rooting culture; (8) domestication and seedling hardening. The method disclosed by the invention has the following beneficial effects: (1) through the tea tree tissue culture using the method, the culture medium components controlling differentiation, subculture growth, rooting and the like in tissue culture are determined; the research determines the best optimization selection for the type, concentration and culture conditions of the growth regulation substances, breaks through the technical barrier during the tea tree tissue culture, solves the technical problems of difficult in-vitro regeneration of the tea tree and low genetic transformation efficiency, and realizes a perfect culture effect.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a tea tree detoxification tissue culture method. Background technique [0002] Tea tree is a perennial woody plant, which has the characteristics of long growth cycle, low self-compatibility, short flowering period, difficult to separate linkage traits, and low beneficial mutation rate, which restricts traditional breeding methods. Therefore, genetic transformation is used to develop tea tree varieties. Improvement is increasingly favored by researchers, and tissue culture can provide an ideal recipient for a large number of gene transfers, and has become a common material for studying transgenic plants. However, there are very few successful cases of tea tree genetic transformation research at present, the main reason being that tea tree regeneration in vitro is difficult and the transformation efficiency is extremely low. [0003] The growth rate of te...

AI Technical Summary

Problems solved by technology

Browning, yellowing, glass seedlings, deformed embryos, pollution and many other problems often occur in the process of plant tissue culture, which affect the experimental results and even lead to the failure of the experiment. loss, urgent need to address

[0004] Chinese patent CN 2016101044044 discloses a method for tissue culture of tea tree young leaves. The method is to inoculate the sterilized tea tree young leaf explants on agar-free liquid medium. The 0.5-1cm place is not cut off, and then the explants are inoculated in the liquid medium for dark culture for 3 days and then transferred to the solid medium for dark culture for 11-13 days. The browning rate of the explants processed by this method is 32%, and the germination rate is as high as 68%; the present invention uses the method of combining liquid-solid co-cultivation and dark culture to significantly reduce the browning rate of explants in the tea tree tissue culture process, and significantly improve the germination rate and efficiency, but it cannot completely solve the yellowing , deformity and other issues