CN102599061A - Method for rapid propagating tissue of cornus alba - Google Patents

Abstract

The invention relates to a method for rapid propagating tissue of cornus alba, which is realized by the steps of explant selecting, cleaning, primary culturing, sub-culturing and rooting culturing, wherein a special culture medium is selected for the primary culturing, the sub-culturing and the rooting culturing, so that the purposes of high the propagation coefficient and good the propagation effect are achieved.

Description

A kind of tatarian dogwood tissue culture and rapid propagation method

Technical field

The present invention relates to the gardening field, be specifically related to a kind of tissue culture and rapid propagation method of tatarian dogwood.

Background technology

Red Ramu (Cornus? Alba) is cornaceae dogwood shrub, up to 3 m; bark purple; young branches pale white pubescent, bald net after being waxy white, red and white old branches, scattered white lenticels and slightly protruding circular annular leaf scars.The florescence 6-7 month; The fruit phase 8-10 month.Tatarian dogwood is the dried seeds of sight of using in the gardens, and autumn leaves is scarlet, and fruitlet is pure white, and fallen leaves backs limb is red gorgeous like coral, is rare sight stem plant, in the gardens on many group planting lawn or alternate plantation with aiphyllium, and effect that must the green and the end setting off each other.The tatarian dogwood seed oil content is about 30%, can supply industrial.In recent years, the demand for tatarian dogwood in the park and garden construction increases gradually, but its breeding practice mainly concentrates in sowing, cuttage, the propagation by layering, and the too much research about tissue-culturing rapid propagation is not arranged, and has influenced it and has applied speed.

Summary of the invention

The technical problem that the present invention mainly solves provides a kind of method for quickly breeding of tatarian dogwood, to remedy in the gardens demand endlessly for the tatarian dogwood resource.

For solving the problems of the technologies described above, the objective of the invention is to realize like this:

A kind of tatarian dogwood tissue culture and rapid propagation method is characterized in that, may further comprise the steps:

Step (1) explant selection: in the first tenday period of a month in May, the clip tatarian dogwood has just been extracted tender tip stem section soon out from the maternal plant;

Step (2) explant is handled: the alcoholic solution with 75% soaks 60s, and aseptic water washing 3 times drains, and puts into 0.1% HgCl again ₂Sterilization is 5 minutes in the solution, and aseptic water washing 3 times cuts stem apex again, and is subsequent use;

Step (3) initial culture: with the stem apex that step (2) cuts, be seeded on the initial culture base, carry out initial culture, the initial culture based component is MS+GA ₃0.4mg/l+BA2mg/l+IAA 0.6mg/l+ 2% sucrose; Initial culture intensity of illumination 800Lux, 22 ± 2 ℃ of cultivation temperature, incubation time 30 ± 2 days makes the stem apex base portion form the bud of growing thickly on every side gradually, when the bud clump grows to 3cm, is cut into single bud, and is subsequent use;

Step (4) subculture: the single bud immigration that step (3) is downcut is equipped with in the airtight container of subculture medium, carries out successive transfer culture; The successive transfer culture based component is MS+ GA ₃0.6mg/l+BA 0.5mg/l+IAA 0.15mg/l+2% sucrose; 23 ± 2 ℃ of cultivation temperature, light application time 16h/d, medium pH value 5.8～6.0 was cultivated 35 days, formed the long budling with 4-5 stipes of 6cm, was cut into the segment of band stipes, and each budling is divided into 2 sections; Per 35 days subcultures like this once expand numerous test-tube plantlet;

Step (5) culture of rootage: step 4 is expanded numerous test-tube plantlet take out, move into and be equipped with in the closed container of root media, carry out culture of rootage, the culture of rootage based component is 1/2MS+ GA ₃0.5mg/l+IBA 0.5mg/l+BA 0.01mg/l+10% sucrose; Test-tube plantlet intensity of illumination in root media is 3000Lux, and 25 ± 2 ℃ of cultivation temperature are cultured to 20 days, and when test-tube plantlet root in medium grew to the new root of 0.5cm, bottle outlet was transplanted.

The invention has the beneficial effects as follows: the tissue-culturing rapid propagation seedling-cultivating method of tatarian dogwood of the present invention reaches about 4 tatarian dogwood test-tube plantlet monthly increment coefficient, can reach the test-tube plantlet about 2～3 strains from explant propagation in 1 year.So both protect maternal plant, can satisfy the large-scale production demand of tatarian dogwood again.

Embodiment

Set forth in detail in the face of preferred embodiment of the present invention down, thereby protection scope of the present invention is made more explicit defining so that advantage of the present invention and characteristic can be easier to it will be appreciated by those skilled in the art that.

The embodiment of the invention comprises:

A kind of tatarian dogwood tissue culture and rapid propagation method is characterized in that, may further comprise the steps:

Step (1) explant selection: the first tenday period of a month in May, in fine day from maternal plant on the clip tatarian dogwood just extracted tender tip stem section soon out;

Step (2) explant is handled: the stem section of choosing is washed 5min earlier in 1% washing powder water, flowing water flushing 15min under water tap again is with the gauze moisture that exhausts; Soak 60s as for the alcoholic solution with 75% on the sterile working platform; Aseptic water washing 3 times drains, and puts into 0.1% HgCl again ₂Sterilization is 5 minutes in the solution, aseptic water washing 3 times, and each 2min, gauze blots surface moisture, cuts stem apex again, and is subsequent use;

Step (3) initial culture: with the stem apex that step (2) cuts, be seeded on the initial culture base, carry out initial culture, the initial culture based component is MS+GA ₃0.4mg/l+BA2mg/l+IAA 0.6mg/l+ 2% sucrose; Initial culture intensity of illumination 800Lux, 22 ± 2 ℃ of cultivation temperature, incubation time 30 ± 2 days makes the stem apex base portion form the bud of growing thickly on every side gradually, when the bud clump grows to 3cm, is cut into single bud, and is subsequent use;

Step (4) subculture: the single bud immigration that step (3) is downcut is equipped with in the airtight container of subculture medium, carries out successive transfer culture; The successive transfer culture based component is MS+ GA ₃0.6mg/l+BA 0.5mg/l+IAA 0.15mg/l+2% sucrose; 23 ± 2 ℃ of cultivation temperature, light application time 16h/d, medium pH value 5.8～6.0 was cultivated 35 days, formed the long budling with 4-5 stipes of 6cm, was cut into the segment of band stipes, and each budling is divided into 2 sections; Per 35 days subcultures like this once expand numerous test-tube plantlet later on;

Step (5) culture of rootage: the test tube seedling and propagating of treating step (4) takes out expanding numerous test-tube plantlet after some, moves into and is equipped with in the closed container of root media, carries out culture of rootage, and the culture of rootage based component is 1/2MS+ GA ₃0.5mg/l+IBA 0.5mg/l+BA 0.01mg/l+10% sucrose; Test-tube plantlet intensity of illumination in root media is 3000Lux, and 25 ± 2 ℃ of cultivation temperature are cultured to 20 days, and when test-tube plantlet root in medium grew to the new root of 0.5cm, bottle outlet was transplanted.The transplanting and management technology is identical with conventional woody tissue cultivating seedling.

The above is merely embodiments of the invention; Be not so limit claim of the present invention; Every equivalent structure or equivalent flow process conversion that utilizes description of the present invention to do; Or directly or indirectly be used in other relevant technical fields, all in like manner be included in the scope of patent protection of the present invention.

Claims (1)

1. a tatarian dogwood tissue culture and rapid propagation method is characterized in that, may further comprise the steps:

Step (1) explant selection: in the first tenday period of a month in May, the clip tatarian dogwood has just been extracted tender tip stem section soon out from the maternal plant;

Step (2) explant is handled: the alcoholic solution with 75% soaks 60s, and aseptic water washing 3 times drains, and puts into 0.1% HgCl again ₂Sterilization is 5 minutes in the solution, and aseptic water washing 3 times cuts stem apex again, and is subsequent use;

Step (3) initial culture: with the stem apex that step (2) cuts, be seeded on the initial culture base, carry out initial culture, the initial culture based component is MS+GA ₃0.4mg/l+BA2mg/l+IAA 0.6mg/l+ 2% sucrose; Initial culture intensity of illumination 800Lux, 22 ± 2 ℃ of cultivation temperature, incubation time 30 ± 2 days makes the stem apex base portion form the bud of growing thickly on every side gradually, when the bud clump grows to 3cm, is cut into single bud, and is subsequent use;

Step (4) subculture: the single bud immigration that step (3) is downcut is equipped with in the airtight container of subculture medium, carries out successive transfer culture; The successive transfer culture based component is MS+ GA ₃0.6mg/l+BA 0.5mg/l+IAA 0.15mg/l+2% sucrose; 23 ± 2 ℃ of cultivation temperature, light application time 16h/d, medium pH value 5.8～6.0 was cultivated 35 days, formed the long budling with 4-5 stipes of 6cm, was cut into the segment of band stipes, and each budling is divided into 2 sections; Per 35 days subcultures like this once expand numerous test-tube plantlet;

Step (5) culture of rootage: step 4 is expanded numerous test-tube plantlet take out, move into and be equipped with in the closed container of root media, carry out culture of rootage, the culture of rootage based component is 1/2MS+ GA ₃0.5mg/l+IBA 0.5mg/l+BA 0.01mg/l+10% sucrose; Test-tube plantlet intensity of illumination in root media is 3000Lux, and 25 ± 2 ℃ of cultivation temperature are cultured to 20 days, and when test-tube plantlet root in medium grew to the new root of 0.5cm, bottle outlet was transplanted.