Methods for callus suspension culture and protoplast separation of camellia oleifera

A suspension culture, protoplast technology, applied in biochemical equipment and methods, tissue culture, cell culture medium, etc., can solve the problems of extensive cultivation management, uneven quality of camellia oleifera varieties, lack of resistant varieties, etc., and achieve easy suspension cell lines. , complete shape, high yield effect

Active Publication Date: 2020-09-25
CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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  • Abstract
  • Description
  • Claims
  • Application Information

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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.

Method used

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  • Methods for callus suspension culture and protoplast separation of camellia oleifera
  • Methods for callus suspension culture and protoplast separation of camellia oleifera
  • Methods for callus suspension culture and protoplast separation of camellia oleifera

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Embodiment 1

[0033] (1) Collect the half-lignified 5-7cm long stems of ‘Huashuo’ Camellia oleifera in the sunny morning between March and May, remove the leaves, rinse with tap water for 20 minutes, and drain. In the ultra-clean workbench, wash twice with sterile water, then disinfect with 75% (v / v) alcohol for 30 seconds, and then use 0.1% (m / v) HgCl 2 The solution was sterilized for 15 minutes, and finally washed with sterile water for 4 to 5 times, the water on the surface of the stem section was blotted with filter paper, cut into 2-3 cm long single-bud stem sections with a sterile blade, and inoculated into the induction medium for induction. After cultivating for 5 days at a temperature of 28°C and in the dark, transfer to light conditions (light intensity is 2000lx, light time 16h) for cultivation, and after 30 to 40 days, compact calluses are induced at the base of the stem segment (see figure 1 -B). Described induction medium is the MS medium that contains 30g / L sucrose, 7g / L aga...

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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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04C12N5/02
CPCC12N5/04C12N5/0025
Inventor 肖诗鑫叶天文李素芳徐新
Owner CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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