Methods for callus suspension culture and protoplast separation of camellia oleifera

Abstract

The invention discloses methods for callus suspension culture and protoplast separation of camellia oleifera and belongs to the field of plant tissue culture. According to the method for callus suspension culture of the camellia oleifera, a massive number of stem segments are taken as explants, compact callus obtained through induction is subjected to subculture on a culture medium containing NAA,2,4-D and 6-BA, and loose granular callus easily establishing a suspension cell system is obtained. By means of the method, the problems that induced callus of the stem segments of the camellia oleifera in the prior art is relatively compact and is not suitable for suspension cell culture are solved. According to the method for protoplast separation of the camellia oleifera, callus protoplast ofthe camellia oleifera is suspended in a sucrose solution with higher density, and impurities such as chips with larger density sink and the protoplast of the camellia oleifera floats through centrifugation, so that separation and purification of protoplast are realized. By means of the method, the problem that the callus protoplast of the camellia oleifera cannot be collected and purified throughcentrifugation is solved, the purification efficiency of the method is high, and the obtained protoplast is clear, visible and high in purity.

Description

technical field [0001] The invention belongs to the field of plant tissue culture, in particular to a method for suspension culture of camellia oleifera callus and separation of protoplasts. Background technique [0002] Camellia oleifera is an important woody oil tree species in my country. It has a cultivation history of more than 1,000 years. It is mainly distributed in the Yangtze River Basin and the areas south of it. It plays an important role in maintaining national grain and oil security, developing rural economy and improving the ecological environment. Camellia oleifera has good economic value. The camellia oil extracted from its seeds has high nutritional value and is rich in unsaturated fatty acids and vitamins. The tea meal left after the oil is extracted from the seeds contains tea saponins, flavonoids, Tannins and other bioactive ingredients have anti-inflammatory, anti-oxidant, antibacterial and other functions. [0003] However, the current development of C...

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Problems solved by technology

[0003] However, the current development of Camellia oleifera industry in my country is facing many problems: (1) Variety of good and bad varieties of Camellia oleifera, resulting in low yield; (2) Lack of varieties resistant to major diseases of Camellia oleifera such as anthracnose; (3) Extensive cultivation and management

And the study of Camellia oleifera callus suspension culture and protoplast isolation has not been reported yet.

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