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Pulsatilla tongkangensis somatic embryogenesis and plant regeneration method

A technology of embryogenesis and pulsatilla, which is applied in the biological field, can solve the problems of difficult transplanting, low survival rate, and large market demand, and achieve the effects of fast reproduction, stable organ differentiation, and consistent genetic traits

Inactive Publication Date: 2015-04-29
LINYI UNIVERSITY
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, Dongjiang Pulsatilla is still in a wild state, the amount of artificial seeding and cultivation is very small, and it is difficult to transplant, and the survival rate is low; in addition, Dongjiang Pulsatilla is a rare ornamental plant with a large market demand

Method used

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  • Pulsatilla tongkangensis somatic embryogenesis and plant regeneration method
  • Pulsatilla tongkangensis somatic embryogenesis and plant regeneration method

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Embodiment 1

[0017] 1. Material preparation

[0018] The material is wild Pulsatilla pulsatillae, and the explants are young leaves.

[0019] 2. Surface disinfection of plant material

[0020] Rinse the leaves of Pulsatilla pulsatillae collected in the wild in tap water for 2 to 4 hours, then carefully wash with washing powder to clean the dust or microorganisms on the leaves, then rinse with sterile water, and then move to the ultra-clean workbench for the next step operate. Soak in 1:3 84 disinfectant solution (containing 1.13-1.53% sodium hypochlorite) added with 0.1% Tween-20 for 15 minutes, then wash with sterile distilled water for 3 to 5 times, and dry the surface moisture with sterilized filter paper. Until there are no water droplets on the surface of the explants, set aside.

[0021] 3. Embryogenic callus and somatic embryo induction

[0022] The sterilized leaves were cut into blocks with an area of ​​0.5 cm × 0.5 cm, cut evenly with a scalpel blade and scored 3 times withou...

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Abstract

The invention provides a pulsatilla tongkangensis somatic embryogenesis and plant regeneration method. The method comprises the following steps: using leaves of pulsatilla tongkangensis as explants, using heteroauxin and zeatin to process the explants to induce somatic embryogenesis, firstly forming embryonic calli on an induction medium by the explants, further differentiating the embryonic calli to form somatic embryos with a spherical structure, a heart-shaped structure and a torpedo-shaped structure, transforming the somatic embryos after bud maturity into mature seedlings with root stocks and leaves. With adoption of the plant tissue culture method, the reproduction speed is fast, the inheritable character is identical, and meanwhile according to the method, the plant regeneration is more stable than organ differentiation; the method provides an important technical support for large-scale vegetative propagation of pulsatilla tongkangensis, and is an effective approach for germplasm resource preservation and protection.

Description

technical field [0001] The invention relates to a somatic embryogenesis and plant regeneration method of Dongjiang Pulsatilla pulsatillae, belonging to the field of biotechnology. Background technique [0002] Pulsatilla tongkangensis (Pulsatilla tongkangensis Y.N.Lee&T.C.Lee) is a perennial herb that grows in the crevices of some cliffs in Yeongwol County and Jeongseon County, Gangwon-do, South Korea. It belongs to Ranunculaceae and is a new species of Pulsatilla. Compared with Korean Pulsatilla, Dongjiang Pulsatilla has larger leaves, shorter petioles and flower stalks, and the flower stalks grow upright without bending, and bloom upwards. Most of Dongjiang Pulsatilla flowers are mainly dark purple, but there are also light purple, lotus root, light pink, purple red and beige, so they have great ornamental value. Because the growth limit of Dongjiang Pulsatilla is limited to the limestone zone in the alpine area, the requirements for the growth environment are relatively ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 廉玉姬林光哲
Owner LINYI UNIVERSITY
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