Method for directly obtaining plants through eggplant anther culture and medium

An eggplant flower and solid culture medium technology, which is applied in the field of plant tissue culture, can solve the problems of not fully satisfying haploid plants, low callus differentiation frequency, low embryoid induction frequency, etc., and achieves overcoming genotype obstacles, Avoid the effect of ploidy confounding and few steps

Active Publication Date: 2017-03-22
北京市海淀区植物组织培养技术实验室 +1
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Problems solved by technology

[0005] At present, eggplant anther culture mostly undergoes two stages of dedifferentiation to form callus and callus differentiation to embryoid bodies or adventitious buds. It needs to be cultured on two different media. The differentiation frequency of tissues is low; free microspore culture can directly obtain embryoid bodies, but the method is cumbersome, and the induction frequency of embryoid bodies is low, which cannot fully meet the production requirements for haploid plants

Method used

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  • Method for directly obtaining plants through eggplant anther culture and medium
  • Method for directly obtaining plants through eggplant anther culture and medium

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Example 1: Anther culture of purple sepal purple eggplant:

[0026] During May-June 2015, the flower buds were retrieved from the greenhouse, and the pollen was stained with DAPI (4,6-diamidino-2-phenylindole) to confirm that the developmental stage was the mononuclear marginal stage. The flower buds were sterilized with 75% alcohol for 1 minute, 5% sodium hypochlorite for 5 minutes, rinsed with sterile water for 3 times, and blotted dry with sterile filter paper. Carefully take out the anthers with small tweezers, place them in the solid medium for anther culture, culture in the dark at 37°C for 3 days, then transfer to 25°C under the light to continue the culture; about 30 days, the white embryoid body can be seen from the cracked anther wall sprouted;

[0027] When the embryoid body differentiates into two small cotyledons, it is transferred to the solid medium for rooting of healthy seedlings to continue to grow, to carry out strong seedlings and rooting. A total...

Embodiment 2

[0031] According to the implementation process of Example 1, 13 kinds of eggplants of different genotypes were tested respectively during 2013-2016, wherein 4 eggplant hybrids (A, C, G, J) were repeated experiments, the results are shown in Table 2 As shown, the hybrid A in Table 2 was repeated in 2014 and 2015; the hybrid C was repeated in 2015 and 2016; the hybrid G ​​was repeated in 2014 and 2015; the hybrid J performed repeated experiments in 2015 and 2016:

[0032] Table 2 The results of embryoid body induction of eggplant anther culture with different genotypes

[0033]

[0034]

[0035] It can be known from Table 2 that the frequency of embryoid body induction is generally 20%-40% using the method of the present invention.

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Abstract

The invention provides a method for directly obtaining plant through eggplant anther cultivation as well as a medium thereof, which belong to the technical field of plant tissue culture technique. The method comprises the following steps: 1) selecting fresh buds; 2) disinfecting the surface of bud and acquiring the aseptic anther; 3) inoculating the aseptic anther in the late uninucleate stage in an anther culture solid medium, culturing the aseptic anther for 2-8 days at dark environment at the temperature of 33-37 DEG C, and performing illumination culture for 30-40 days at the temperature of 25-28 DEG C; and 4) transferring the cotyledonous embryoid to a healthy seedling solid medium for continuously culturing the materials, and growing to obtain the plant. The method has the following advantages that the method is simple and easy to carry out, the in-vitro culture of the anther can directly induce the embryoid to obtain the haploid plant, a condition that the callus formed by anther culture induction is transferred to a differentiation medium, and then the plant regeneration is induced can be avoided; and the chromosome ploidy mixing phenomenon due to participation of diploid somatic cells such as the anther wall in callus can be avoided.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method and a culture medium for directly obtaining plants by culturing eggplant anthers. Background technique [0002] Anther culture is an important way to obtain haploid plants. Homozygous diploids can be obtained quickly through this technology, which has the advantages of shortening the breeding cycle, improving selection efficiency, and overcoming the sterility of distant hybrids. It can also be used as a molecular marker and Good material for genetic mapping research. [0003] There are three main ways to obtain eggplant haploid plants by anther culture or free microspore culture: 1. First dedifferentiate to form callus, then induce callus to differentiate to form embryoid, and finally develop from embryoid to complete plant; 2. Adventitious buds are directly differentiated from the induced callus, and the adventitious buds develop into complete ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 邓晓梅张秦惠志明张树根张军民李春玲
Owner 北京市海淀区植物组织培养技术实验室
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