CN101574056A

CN101574056A - Actinidia arguta tissue culturing method

Info

Publication number: CN101574056A
Application number: CNA2008100644662A
Authority: CN
Inventors: 田新华; 邢亚娟; 兰世波; 李海霞; 周志军
Original assignee: FORESTRY RESEARCH INSTITUTE OF HEILONGJIANG PROVINCE
Current assignee: FORESTRY RESEARCH INSTITUTE OF HEILONGJIANG PROVINCE
Priority date: 2008-05-08
Filing date: 2008-05-08
Publication date: 2009-11-11

Abstract

The invention belongs to the field of plant tissue culturing technologies and forest cultivation and relates to a high-efficiency actinidia arguta propagation method. The method comprises four stages of explant sterilization, inoculation, multiplication and rootage. The method solves the important technical problems of microbody quick propagation and root induction of the actinidia arguta, achieves the aims of efficiently inducing explants to separate and propagate and guaranteeing that seedlings survive with high quality, preserves all good characters of a matrix, and ensures stable hereditary character. The method can form large qualities of good test-tube seedlings within a short period and is suitable for large-scale and industrial production.

Description

Actinidia arguta tissue culturing method

Technical field

The present invention relates to the asexual reproduction method of a kind of berry of the Northeast, i.e. the tissue culture method of tara vine.The invention belongs to plant tissue culture technique and forest and cultivate the field.

Background technology

The tara vine fruit is a kind of rare, natural, green edible berry, because of its potential nutritive value and commercial value, being rare, pure natural, the green edible berry in a kind of world, is one of third generation fruit that has DEVELOPMENT PROSPECT.Yet owing to there is not the harvesting of plan predation formula, cause the tara vine resource to be seriously damaged, natural repair ability reduces, and reserves and Ke Cai amount fall sharply, and imbalance between supply and demand increasingly sharpens.

The seedling growing process of China tara vine mainly is vegetative propagation technology such as (being plant division, press strip, cuttage, green wood cutting), and it is few to utilize tissue culture technology to breed the data of tara vine.Consult a large amount of related datas and find to have only Liu Ruilin once to deliver the bulletin of tissue culture aspect in calendar year 2001.But tara vine tissue culturing system does not set up as yet, and popularizing as yet of improved seeds do not finished, and the nursery stock disparities between supply and demand are increasingly sharpened, and has had a strong impact on the development of tara vine deep processing industry.My institute had carried out systematically research to tara vine tissue culturing system in 2007.

Summary of the invention

The present invention adopts tissue culture technique to tara vine, with the spray of sprouting after the resting shoot water planting is explant, through explant sterilization, the tender shoots inoculation, the screening of differentiation and proliferation medium, steps such as root media screening, the nutritional need of test-tube plantlet in the research incubation, hormone is to the regulation and control of seedling growth differentiation, solve the microbody fast breeding of tara vine, the important technological problems of root induction, set up perfect tara vine tissue culturing system, reached and efficiently induced explant differentiation and proliferation and high-quality to guarantee the purpose that seedling survives, and whole merits of parent have been preserved, stabilization characteristics of genetics.This method can form a large amount of good test-tube plantlets in a short time, carries out scale, batch production production.

Description of drawings

10 days growing way situation of Fig. 1 tara vine water planting tender shoots inoculation, mounted blade, the leaf look dark green

Fig. 2 a section processes shoot proliferation is cultivated the test-tube plantlet of differentiation in 25 days and is counted 25 days test-tube plantlet heights of seedling of b different disposal shoot proliferation cultivation

Four kinds of Fig. 3 a tara vines take root for handling and cultivated 30 days, have only to handle 1 and do not see and take root that all the other 3 processing are all taken root, and 4 the situation of taking root wherein handled is best, and rooting rate reaches 87.0%.

Embodiment

The spray of sprouting after water planting with the tara vine resting shoot is an explant, specifically is made up of following four steps:

(1) explant sterilization at first adopts conventional method that the tender shoots of water planting extraction is cleaned with washing powder solution, with 0.1% mercuric chloride sterilization 5 minutes, soaks 40 minutes with sterile water then;

(2) be seeded under the aseptic condition tender shoots is seeded on the initial culture base, this medium is to have added the 6-benzyladenine (6-BA) of 0.5mg/L, the gibberellin (GA of 1mg/L in the conventional medium of MS ₃), PH 5.8, cultivate basic seedling.Cultivation temperature is 25 ± 2 ℃, periodicity of illumination 12h/d, luminous intensity is 2000lux, wait to cultivate 20 days after, the about 3～4cm of height of seedling promptly begins the next stage;

(3) shoot proliferation is transferred to basic seedling and is carried out enrichment culture on the shoot proliferation medium.Screen with MS+6-BA (0.5mg/L, 1mg/L)+ZT (0.5mg/L, 1mg/L) or 8 kinds of medium of IAA (0.5mg/L, 1mg/L), cultivation temperature is 25 ± 2 ℃, periodicity of illumination 12h/d, and luminous intensity is 2000lux, cultivates 30 days.Optimum multiplication medium is to have added the 6-benzyladenine (6-BA) of 0.5mg/L, the zeatin (ZT) of 1mg/L in the conventional medium of MS as a result; Cultivated 25 days, the propagation multiple reaches 4.0;

(4) take root length in the clip enrichment culture is that the healthy seedling of 1.5～2.0cm is inoculated on four kinds of root medias, adopts the single factor experiment design to carry out the culture of rootage screening.Cultivation temperature is 25 ± 2 ℃, periodicity of illumination 12h/d, and luminous intensity is 2000lux.The result has added the medium rooting efficiency of indolebutyric acid (IBA) of 0.2mg/L in the MS medium best, reaches 87.0% cultivating 30 days rooting rates.

Claims

The method for tissue culture of tara vine (Actinidia arguta) is characterized in that being made up of following four steps:

1. explant sterilization adopts conventional method that the tender shoots of water planting extraction is cleaned with washing powder solution, sterilizes 5 minutes with 0.1% mercuric chloride in super quiet workbench then, soaks 40 minutes with sterile water;
2. be seeded under the aseptic condition tender shoots is seeded on the initial culture base, this medium is to have added the 6-benzylaminopurine (6-BA) of 0.5mg/L, the gibberellin (GA of 1mg/L in the conventional medium of MS ₃), PH 5.8, cultivation temperature is 25 ± 2 ℃, periodicity of illumination 12h/d, luminous intensity is 2000lux, wait to cultivate 20 days after, the about 3～4cm of height of seedling promptly begins the next stage;
3. shoot proliferation is transferred to the nursery stock of gained on the proliferated culture medium and is bred, this medium is to have added the 6-benzyladenine (6-BA) of 0.5mg/L, the zeatin (ZT) of 1mg/L in the conventional medium of MS, PH 5.8, cultivation temperature is 25 ± 2 ℃, periodicity of illumination 12h/d, luminous intensity is 2000lux, cultivates 25 days, and the propagation multiple reaches 4.0;
4. the length in the clip enrichment culture of taking root is that the healthy seedling of 1.5～2.0cm is inoculated into root media, this medium is in the MS medium, added the indolebutyric acid (IBA) of 0.2mg/L, PH 5.8, cultivation temperature is 25 ± 2 ℃, periodicity of illumination 12h/d, luminous intensity is 2000lux, rooting rate reached 87.0% in 30 days.