Camellia sinensis flower protoplast, preparation method therefor and application thereof

Abstract

The invention discloses a tea tree flower protoplast, its preparation method and application. The tea tree flower is placed in a digestive enzyme solution to obtain the protoplast of the tea tree flower after enzymolysis and digestion. The invention can completely degrade 100% of the tea tree flowers by adjusting the formula system of the digestive enzyme liquid, and obtain a large amount of undamaged protoplasts of the tea tree flowers. At the same time, the expression rate obtained after the exogenous gene is transformed into protoplasts of Camellia sinensis can reach more than 30%, which is consistent with the average expression level in the model plant Arabidopsis thaliana. The present invention provides more direct evidence for future tea tree gene function verification, subcellular localization and metabolite detection, which is more targeted than verification in model plants such as Arabidopsis and tobacco; Compared with the transient expression experiment of injecting tobacco or tissue cultured tea seedlings, the equipment and materials required by this method are simple, fast and effective, and the experimental results 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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