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Method for acquiring regeneration plant by Camellia chekiangoleosa Hu anther culture

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, difficulty in obtaining homozygous plants, low regeneration efficiency, etc., so as to improve the callus differentiation rate. , the effect of shortening the breeding cycle and speeding up the breeding process

Active Publication Date: 2019-12-24
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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  • Method for acquiring regeneration plant by Camellia chekiangoleosa Hu anther culture
  • Method for acquiring regeneration plant by Camellia chekiangoleosa Hu anther culture
  • Method for acquiring regeneration plant by Camellia chekiangoleosa Hu anther culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] This embodiment provides a method for in vitro culture of Zhejiang Red Camellia anthers to obtain regenerated plants, including the following steps:

[0046] (1) Selection of flower buds

[0047] On 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 recovered flower buds were divided into 5 groups according to the size of the transverse diameter, the buds were peeled off, and the colors of pollen grains of different groups were observed, and then the anthers were stained with acetic acid 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, select the corresponding flower buds whose microspores are in the uninucleate marginal stage for subsequent culture.

[0048] (2) Bud low temperature pretreatment

[0049] Put the f...

Embodiment 2

[0065] This example provides a method for in vitro culture of Zhejiang Red Camellia anthers to obtain regenerated plants. The steps are the same as Example 1, the difference is that the conditions of individual steps are different, and the culture medium used is different. This example The different conditions and culture medium adopted are specifically referred to as follows, and the experimental steps and conditions not clearly stated can be found in Example 1:

[0066] (2) Bud low temperature pretreatment

[0067] Low temperature treatment for 5 days.

[0068] (3) Disinfection of explants

[0069] Soak the flower buds with detergent for 15 minutes (intermittent agitation), using 0.2% (m / m) carbendazim (50% active ingredient) and 1% (m / m) polyoxin (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 is: MS minimal medium (powder), 1mg / L ...

Embodiment 3

[0077] This example provides a method for in vitro culture of Zhejiang Red Camellia anthers to obtain regenerated plants. The steps are the same as Example 1, the difference is that the conditions of individual steps are different, and the culture medium used is different. This example The different conditions and culture medium adopted are specifically referred to as follows, and the experimental steps and conditions not clearly stated can be found in Example 1:

[0078] (2) Bud low temperature pretreatment

[0079] Low temperature treatment for 7 days.

[0080] (3) Disinfection of explants

[0081] Soak the flower buds with detergent for 20 minutes (intermittent agitation), using 0.2% (m / m) carbendazim (50% active ingredient) and 1% (m / m) polyoxin (1% active ingredient) Soak for 15 minutes (stir with a brush intermittently during the soaking process), continue to rinse with tap water for 4 hours; disinfect the cleaned flower buds with 75% (v / v) ethanol for 60 seconds, wash...

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Abstract

The invention belongs to the field of plant cell engineering, and particularly relates to a method for acquiring a regeneration plant by Camellia chekiangoleosa Hu anther culture. The method includesthe steps: low-temperature pretreatment of Camellia chekiangoleosa Hu anthers; callus tissue induction culture; differentiation culture; rooting culture. Callus tissue induction culture is implementedby the aid of a dark culture mode, a callus tissue induction culture medium used by callus tissue induction culture comprises a hormone combination, anti-browning agents and 30-80g / L sucrose, and thehormone combination includes a combination of 2, 4-D, KT and TDZ or a combination of 2, 4-D, 6-BA and TDZ. According to the method, the callus induction rate, the differentiation rate and the rootingrate of regeneration culture of the Camellia chekiangoleosa Hu anthers can be remarkably increased.

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 through in vitro culture of anthers of Camellia japonica. Background technique [0002] Red camellia mainly comes from the species and varieties of Sect Camellia in the family Theaceae (Theaceae) Camellia. The flowers are red, gorgeous, and have high ornamental value. At the same time, some species have a high rate of seed setting and oily kernels. The rate is high, and it can also be used as a high-quality woody edible oil. [0003] Zhejiang red camellia (Camellia chekiangoleosa Hu), also known as Zhejiang safflower camellia, 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 large, bright and dense. The fruit resembles an apple, and its color is bright red or yellow-green. When it matures, the...

Claims

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Application Information

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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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