Camellia sinensis flower protoplast, preparation method therefor and application thereof

Abstract

The invention discloses a camellia sinensis flower protoplast, a preparation method therefor and an application thereof. Camellia sinensis flowers are put into a digestive enzyme solution and are enzymatically hydrolyzed and digested to obtain the camellia sinensis flower protoplast. By adjusting a formula system of the digestive enzyme solution, the camellia sinensis flowers can be degraded by 100% and a large amount of unbroken camellia sinensis flower protoplasts can be obtained. Meanwhile, the expression rate obtained after the camellia sinensis flower protoplast is converted by an exogenous gene can reach more than 30% and is consistent with the average expression level in arabidopsis thaliana as a model plant. More direct evidences are provided for camellia sinensis gene function verification and even subcellular localization as well as metabolite detection in the future, and the verification is more targeted in comparison with that in the model plants such as the arabidopsis thaliana, tobaccos and the like; and, compared with a transient expression experiment of agrobacterium injected tobaccos or tissue culture tea seedlings for more than one week, devices and materials required in the method are simple, the method is quick and effective, and an experimental result can be seen in two days.

Description

technical field [0001] The invention belongs to the technical field of cell biology, and in particular relates to a tea tree flower protoplast and its preparation method and application. Background technique [0002] Tea is the main drink in China and even in the world. The reason why it has become the main drink selected by the Chinese for thousands of years and one of the seven things in life is that tea contains a variety of functional ingredients, which can promote human metabolism, vitality, Improving immunity, promoting body fluid and activating blood circulation, concentrating and meditating, modern science has discovered that it has many effects such as anti-cancer, anti-inflammation, lowering blood sugar, and lowering blood lipids. Tea has so many functional substances, but the current research on them is still shallow, and the metabolism and regulation mechanism of these substances are still unclear. This is because these substances are unique to tea, and there is ...

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Problems solved by technology

Tea has so many functional substances, but the current research on them is still shallow, and the metabolism and regulation mechanism of these substances are still unclear. This is because these substances are unique to tea, and there is no model plant for reference. On the other hand, because of the specificity of this species, there is no good expression system to verify the function of a certain protein gene, and many mechanisms cannot be obtained because there are no related metabolic pathways in Arabidopsis or tobacco. verification; and tea is a woody plant, and the development of its transgenic technology and tissue culture technology is quite difficult. Therefore, the development of tea tree protoplast isolation and expression technology is an important breakthrough direction

At present, there are protoplast isolation techniques for some woody plant leaves, such as apples, oranges, etc. However, due to the high content of cellulose and polyphenols in the leaves of tea trees, it is difficult to obtain a large number of protoplasts in good condition for later transformation and expression experiments

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