CN101069487A

CN101069487A - Labbiteye blueberry tissue culturation method

Info

Publication number: CN101069487A
Application number: CNA2007100245028A
Authority: CN
Inventors: 张长青; 朱土农
Original assignee: Jinling Institute of Technology
Current assignee: Jinling Institute of Technology
Priority date: 2007-06-20
Filing date: 2007-06-20
Publication date: 2007-11-14

Abstract

The present invention belongs to the field of plant tissue culture technology, in the concrete, it relates to a high-effective breeding method of rabbiteye blueberry. Said method includes four stages of induction, proliferation, rooting and selecting seedling.

Description

Labbiteye blueberry tissue culturation method

One technical field

The invention belongs to field of plant tissue culture technique, be specifically related to the method for plant tissue culture of lagophthalmos blueberry.

Two research backgrounds

The lagophthalmos blueberry is the treasure in the fruit tree, and unique flavor can be eaten raw, also can process various lagophthalmos blueberry series foods, can also be as the raw material that extracts anthocyanidin.Anthocyanidin in its fruit is a kind of water-soluble biological flavones, has pre-myopia prevention, strengthens cardiac function, anticancer, anti-cardiovascular disease effect.International food and agricultural organization has classified it as one of human five big health foods.The tradition cuttage breeding method exists the difficulty of taking root, scarce limit that reproduction coefficient is low.At present, both at home and abroad about the research of lagophthalmos blueberry tissue culture system not also, efficient breeding of method also lacks very much.

Three summary of the invention

The problem that the present invention need solve provides a kind of efficient method for tissue culture of lagophthalmos blueberry.Technical scheme is:

1. induce

Lagophthalmos blueberry stem section is seeded on the inducing culture.This medium is the zeatin that has added 10 mg/litre in the conventional medium of WPM.Cultivate after 1 month, treat that culture grows young sprout after, promptly begin the next stage.

2. breed

The young sprout of gained in the 1st step cut transfer to proliferated culture medium.This medium is the zeatin that has added 5 mg/litre in the conventional medium of WPM.Cultivate after 1 month, use again with quadrat method subculture 3 times.Entered for the 3rd step then.

3. take root

With the seedling of the 2nd step gained, transfer in the conventional root media.After one month, root system appears in the seedling base portion.After 15 days, enter next step.

4. hardening

The seedling of taking root that the 3rd step was obtained is planted in the hardening matrix.This matrix constituent is the peat composed of rotten mosses and quartz sand, and the peat composed of rotten mosses and quartz sand ratio are 1-3: 1.Can obtain a large amount of finished product seedlings after requiring to manage according to general hardening.

The present invention has adopted three kinds of special matrix, i.e. inducing culture, proliferated culture medium and hardening matrix.The zeatin concentration that inducing culture adds is 10 mg/litre; The zeatin concentration that proliferated culture medium adds is 5 mg/litre; Hardening matrix is 1-3: 1 the peat composed of rotten mosses and quartz sand combination.

Adopt this method, just can see at induction period that bud sprouted in general 3 days, 70% bud sprouting was promptly arranged in 7 days.After 20 days, new-tip length can reach 0.8-1.2 centimetre, blade quantity 6-8 sheet, and size is close to 1/3 (Fig. 1) of the normal leaf in field.Simultaneously, tangible white powder can occur on stem section and blade, the leaf look yellowish green, fresh and tender, and expand stem section bottom.In the multiplicative stage, can see that regrowth began growth and differentiating phenomenon in 4 days, and differentiation come out newly slightly also obviously sturdy, the rate of increase can reach (Fig. 2) more than 12 times.In the hardening stage, the amount of growth that survival rate reached as high as after 99%, month reaches more than 1.2 centimetres, newly-increased blade 2-4 sheet (Fig. 3).

The invention has the beneficial effects as follows: 1. significantly improved the germination rate of explant and greater than 1 centimetre branch number; 2. the young sprout rate of increase has been reached more than 12 times; 3. the survival rate of seedling has reached 99%.

The invention solves lagophthalmos blueberry tradition cottage method take root difficulty, a difficult problem that reproduction coefficient is low, a kind of method of suitability for industrialized production high-quality finished product seedling is provided for the development and use of lagophthalmos blueberry.

Lagophthalmos blueberry of the present invention can be developed into lagophthalmos blueberry juice, lagophthalmos blueberry fruit wine, lagophthalmos blueberry jam, lagophthalmos blueberry mixed health food, and prepares medicine antitumor, the treatment angiocardiopathy.

Four description of drawings

The lagophthalmos blueberry stem section of growing on Fig. 1 inducing culture

The lagophthalmos blueberry young sprout of growing on Fig. 2 proliferated culture medium

Lagophthalmos blueberry in Fig. 3 hardening matrix survives seedling

Five embodiments

1. draw materials and sterilize: not lignified young sprout is gathered in the field, behind the flushing with clean water 2h, and alcohol disinfecting 30 seconds, aseptic water washing 3 times.Sterilized 10 minutes aseptic water washing 3 times more again with 0.1% mercuric chloride.Cut off the blade size 2/3 after, branch is cut into the sections of 1-2 bud.

2. inducing culture

On superclean bench, the stem section that shears is seeded on the inducing culture.This medium consists of WPM+10 mg/litre zeatin.

3. enrichment culture

Can obtain length after 30 days at a large amount of young sprouts more than 1 centimetre.With the young sprout that grows move be connected on begin on the proliferated culture medium propagation.The zeatin that consists of the WPM+5 mg/litre of proliferated culture medium.Subculture 3 times.

4. take root

The stem section that previous step obtains is taken out, be inoculated into and carry out culture of rootage on the root media.Used prescription is the heteroauxin of 1/2 WPM+0.1 mg/litre.

5. hardening

Opened bottle cap in advance in 3 days.During transplanting, carefully take out seedling, wash clean with tweezers.Seedling that selection is taken root and the ratio of being transplanted to the peat composed of rotten mosses and quartz sand are 1-3: in 1 the matrix, spend 15 days in the group training is indoor in the plastic greenhouse of complete closed.Keep humidity 100% in the canopy therebetween.Move into the field afterwards, and in the plastic greenhouse of sealing, spend 5 days under maintenance humidity 80%, light transmittance 70% situation, begin to leak informaton.After 15 days, throw off the canopy film.

Claims

1, Labbiteye blueberry tissue culturation method is characterized in that being made up of following three steps:

(1) induces

Lagophthalmos blueberry stem section is seeded on the inducing culture, and this medium is the zeatin that has added 10 mg/litre in the conventional medium of WPM, cultivates after 1 month, treat that culture grows young sprout after, promptly begin the next one stage;

(2) propagation

The young sprout of gained in the 1st step cut to transfer on the proliferated culture medium breed, this medium is the zeatin that has added 5 mg/litre in the conventional medium of WPM, cultivates after 1 month, again young sprout is continued to breed, and obtains to enter for the 3rd step behind a large amount of young sprouts;

(3) take root

With the young sprout of the 2nd step gained, transfer in the conventional root media, after one month, root system appears in the young sprout base portion, after 15 days, carries out hardening.

2, the hardening off method of the seedling of taking root of turning out according to claim 1 cultural method is characterized in that Routine Management is carried out in the transplantation of seedlings of taking root that claim 1 is turned out in hardening matrix, and the constituent of this hardening matrix is the peat composed of rotten mosses and quartz sand, and ratio is 1-3: 1.

3, the lagophthalmos blueberry of cultivating according to claim 1 cultural method is preparing lagophthalmos blueberry juice, lagophthalmos blueberry fruit wine, uses in lagophthalmos blueberry jam and the lagophthalmos blueberry mixed health product.

4, the lagophthalmos blueberry of cultivating according to claim 1 cultural method is preparing the application for the treatment of in tumour, the cardiovascular disease medicine.