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A method for in vitro culture of Zhejiang Red Camellia anthers to obtain regenerated plants

A technology for in vitro culture and regeneration of plants, applied in the field of plant cell engineering, can solve the problems of low induction rate, differentiation rate and rooting rate of regeneration culture, low regeneration efficiency, and long cycle

Active Publication Date: 2021-05-14
RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because camellia sponsonii is a perennial woody plant with highly heterozygous genes and rich genetic diversity, the traditional hybrid breeding method has a long cycle, and the induction rate, differentiation rate and rooting rate of its regeneration culture are relatively low, and the regeneration efficiency is low. homozygous plants

Method used

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  • A method for in vitro culture of Zhejiang Red Camellia anthers to obtain regenerated plants
  • A method for in vitro culture of Zhejiang Red Camellia anthers to obtain regenerated plants
  • A method for in vitro culture of Zhejiang Red Camellia anthers to obtain regenerated plants

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] The present embodiment provides a method for obtaining regenerated plants by in vitro culture of Zhejiang red camellia anther, comprising the following steps:

[0046] (1) Selection of flower buds

[0047] Choose a sunny morning in October, pick the buds of Zhejiang red camellia, put them in a ziplock bag, put them in an ice box and bring them back to the laboratory in time. The collected flower buds were divided into 5 groups according to the size of the transverse diameter, the buds were peeled off, the color of the pollen grains in different groups was observed, and then the anthers were stained with acetic magenta, and the developmental stages of the microspores were observed under a microscope. According to the observation results under the microscope, the size and color of the flower buds, the corresponding flower buds with microspores in the mononuclear border stage were selected for subsequent culture.

[0048] (2) Low temperature pretreatment of flower buds

...

Embodiment 2

[0065] This embodiment provides a method for obtaining regenerated plants by in vitro culture of Zhejiang red camellia anther. The steps are the same as those in Embodiment 1, except that the conditions of individual steps are different, and the medium used is different. This embodiment The specific reference to the different conditions and culture medium used are as follows, and the experimental steps and conditions that are not clearly stated are referred to in Example 1:

[0066] (2) Low temperature pretreatment of flower buds

[0067] Low temperature treatment for 5 days.

[0068] (3) Sterilization of explants

[0069] Soak flower buds with dish soap for 15min (intermittent stirring), use 0.2% (m / m) carbendazim (50% active ingredient) and 1% (m / m) polyantimycin (1% active ingredient) Soak in 75% (v / v) ethanol for 50s, 0.1% (m / m) HgCl 2 Surface disinfection for 18 minutes.

[0070] (4) Callus induction culture

[0071] The callus induction medium was: MS basic medium (...

Embodiment 3

[0077] This embodiment provides a method for obtaining regenerated plants by in vitro culture of Zhejiang red camellia anther. The steps are the same as those in Embodiment 1, except that the conditions of individual steps are different, and the medium used is different. This embodiment The specific reference to the different conditions and culture medium used are as follows, and the experimental steps and conditions that are not clearly stated are referred to in Example 1:

[0078] (2) Low temperature pretreatment of flower buds

[0079] Low temperature treatment for 7 days.

[0080] (3) Sterilization of explants

[0081] Soak flower buds with dish soap for 20min (intermittent stirring), use 0.2% (m / m) carbendazim (50% active ingredient) and 1% (m / m) polyantimycin (1% active ingredient) Soak for 15 minutes (intermittently stir with a brush during the soaking process), and continue to rinse with tap water for 4 hours; the cleaned flower buds are disinfected with 75% (v / v) et...

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Abstract

The application belongs to the field of plant cell engineering, and in particular relates to a method for obtaining regenerated plants through in vitro culture of anthers of Camellia japonica. The method for obtaining regenerated plants by in vitro culture of red camellia anthers comprises: low-temperature pretreatment of red camellia anthers, callus induction culture, differentiation culture and rooting culture; wherein, the callus induction culture adopts dark Culture; the callus induction medium adopted comprises: hormone combination, anti-browning agent and the sucrose of 30-80g / L, and described hormone combination is 2,4-D, KT and TDZ combination, or 2,4- D. Combination of 6‑BA and TDZ. The method can significantly improve the callus induction rate, differentiation rate and rooting rate of the red camellia anther regeneration culture.

Description

technical field [0001] The application belongs to the field of plant cell engineering, and in particular relates to a method for obtaining regenerated plants by in vitro culturing of red camellia anthers. Background technique [0002] Black camellia mainly comes from the species and varieties of the SectCamellia group (SectCamellia) of the family Theaceae (Theaceae), with red, gorgeous flowers, and has a very high ornamental value. It can also be used as a high-quality woody edible oil. [0003] Zhejiang red camellia (Camellia chekiangoleosa Hu), also known as Zhejiang safflower oil tea, is a unique species of Camellia spp. in my country, and is an excellent red camellia species. Zhejiang red camellia is cold-resistant and blooms in early spring. The flowers are huge, bright and dense. The fruit resembles an apple, with bright red or yellow-green color. Zhejiang red camellia has high oil value. The oil content of the seeds of Zhejiang red camellia is as high as about 60%,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00A01G31/00A01G24/15A01G24/28
CPCA01G31/00A01H4/00A01H4/001A01H4/008A01G24/15A01G24/28
Inventor 叶思诚姚小华卓仁英王开良马康荣
Owner RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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