Method for generating Japan lawn grass plant by mature embryo calls induction and culture medium

A basic medium and the technology of zoysia grass, which are applied in the field of plant tissue culture and plant regeneration, can solve the problems of difficult long-term subculture and differentiation ability of callus, and achieve long-term subculture preservation and maintain differentiation ability, The effect of good differentiation ability

Inactive Publication Date: 2006-03-22
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The callus produced by these methods is difficult to subculture for a long time and maintain differentiation ability, so it is difficult to be a recipient of genetic transformation

Method used

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  • Method for generating Japan lawn grass plant by mature embryo calls induction and culture medium
  • Method for generating Japan lawn grass plant by mature embryo calls induction and culture medium
  • Method for generating Japan lawn grass plant by mature embryo calls induction and culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: the establishment of Japanese zoysia regeneration system

[0036] 1. The source and treatment of test materials:

[0037]Mature Japanese Zoysia (Zoysia japonica Steud) seeds of the test material of the present invention were purchased from Beijing China Seed Industry Group. In the present invention, induction, subculture, and differentiation of callus of Zoysia japonica are not limited by species.

[0038] Grind Zoysia japonica seeds with distilled water in a mortar for about 15 minutes, rinse under tap water to remove the seed coat, and then disinfect with 76% (V / V) alcohol for 30 seconds on an ultra-clean workbench (or sterile inoculation box) , and then sterilized with 0.1% (W / V) mercuric chloride solution for 20-25 minutes, rinsed with sterile water 5 times, and washed for 3-5 minutes each time to obtain sterilized seeds for processing and set aside.

[0039] 2. Culture medium design:

[0040] Table 1 lists the composition and consumption amount of ...

Embodiment 2

[0049] Example 2: Effects of different concentrations of KT on long-term subculture preservation of callus

[0050] (1) Material:

[0051] A callus in a differentiated state formed after 2-3 generations of callus proliferation and culture, and the callus is subjected to long-term subculture.

[0052] (2) Preparation of callus subculture medium:

[0053] The preparation of callus subculture medium is shown in Table 1. In order to explore the effects of different concentrations of KT on the continuously subcultured Zoysia japonica, in this example the applicant specially added the culture medium shown in Table 2 for comparative experiments.

[0054] 3. Training method:

[0055] The induced compact callus formed a callus in a differentiated state after 2-3 generations of subculture (see Figure 4 shown). Select the yellow-white, granular, and well-dispersed callus around the callus in the differentiated state, or the granular, dry, and hard callus formed around the bud struc...

Embodiment 3

[0058] Embodiment 3: The differentiation rate of the Zoysia japonica callus of continuous subculture on different differentiation medium

[0059] The preparation of medium for callus differentiation is shown in Table 1.

[0060] The long-term preserved callus of the present invention often coexists in multiple states (see Figure 5 ), the black arrow points to the callus selected during preservation, the blue arrow points to the callus with only proliferative ability but no differentiation ability, and the red arrow points to a callus with a higher degree of organization , its production is a sign that the subcultured callus of the present invention has preservation value.

[0061] In this example, applicants used the callus that had been subcultured for 6 generations for the experiment of differentiating shoots and plants. On the basis of the general differentiation medium shown in Table 1, the applicant specially designed the differentiation medium shown in Table 3 to dete...

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Abstract

The present invention relates to plant tissue and cell culture technology. Mature seed of Zoysia japonica Steud as lawn grass is used as explant to induce callus and the callus is proliferated and induced to differentiate to form complete small plant. The present invention includes the following steps: A. culturing mature seed of Zoysia japonica Steud in culture medium containing callus inducing agent and inducing callus; B. inoculating callus in excellent state to proliferation culture medium for proliferation or secondary culture; and C. inoculating proliferated or secondary cultured callus in excellent state to differentiating culture medium to induce complete small plant. Compared with available technology, the present invention has obviously raised Zoysia japonica Steud explant callus inducing rate and plant regenerating efficiency.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture and plant regeneration. Specifically, the invention relates to a method for plant regeneration through callus induction using mature seeds of Zoysia japonica as explants and a culture medium for the method . Background technique [0002] Japanese Zoysia (Zoysia japonica Steud.) is a warm-season grass species that is widely used and has strong adaptability among lawn plants. It has a wide distribution and strong resistance, and plays an important role in modern urban greening, sports field construction and soil and water conservation, but the short green period has become a bottleneck restricting its large-scale application. Using modern biotechnology, such as genetic transformation, protoplast The genetic improvement of Zoysia japonica through fusion, induction of somatic mutants, etc. provides a broad space for the innovation of Zoysia japonica germpl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 包满珠樊晓莉张俊卫刘国锋高丽萍胡惠蓉王文恩
Owner HUAZHONG AGRI UNIV
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