CN103828719A

CN103828719A - Method for tissue culture of liquidambar styraciflua

Info

Publication number: CN103828719A
Application number: CN201410090748.5A
Authority: CN
Inventors: 李在根
Original assignee: HUNAN TIANFU FORESTRY SCIENCE & TECHNOLOGY Co Ltd
Current assignee: HUNAN TIANFU FORESTRY SCIENCE & TECHNOLOGY Co Ltd
Priority date: 2014-03-13
Filing date: 2014-03-13
Publication date: 2014-06-04

Abstract

The invention relates to a tissue culture method for plants, and in particular relates to a method for tissue culture of liquidambar styraciflua. The method comprises the following steps: selecting effective stems with axillary buds of liquidambar styraciflua as an explant, performing pretreatment such as sterilization on the effective stems firstly, subsequently inoculating the explant onto an axillary bud development starting culture medium for culture on an ultra-clean working table so as to obtain sterile seedlings; inoculating the sterile seedlings into a bud proliferation and elongation culture medium for culture, selecting growing points of the proliferated buds, sequentially transferring into a callus induction culture medium, a callus proliferation culture medium and a root induction culture medium for culture according to culture progress so as to obtain non-poisonous plants, and transplanting and planting the non-poisonous plants. According to the method, the unique explant and the formulae of the culture mediums are adopted, the success rate and the plant quality in tissue culture of liquidambar styraciflua are greatly improved, and strong guarantee for industrial development of liquidambar styraciflua is provided, and the method is rapid in culture and economical.

Description

A kind of method for tissue culture of American red-maple

Technical field

The present invention relates to the method for tissue culture of plant, relate in particular to the method for tissue culture of viewing and admiring forest, specifically a kind of method for tissue culture of American red-maple.

Background technology

American red-maple (Acerrubrum) has another name called red maple, the red maple in North America, and Aceraceae maple belongs to.12-18 meters of the height of trees, its tree performance grace, with luxuriant foliage and spreading branches in leafy profusion, blade profile is slim and graceful, and leaf look beautiful, and the earliest, mass-tone is scarlet in variable color in autumn, and parachrome is orange, and look gorgeous as flower, is mainly used in that wooden land is viewed and admired and courtyard greening, is also precious family potted plant seeds.At present, American red-maple generally adopts seminal propagation, propagation by grafiting and three kinds of modess of reproduction of cottage propagation, but these three kinds of mode reproduction speeds are slower, tissue culture quick breeding is a kind of effectively modes of reproduction, it has not only improved the proliferative speed of red maple, also provide strong guarantee to the quality of nursery stock, for afforestation provides sufficient seedling simultaneously.

Near the virus concentration of White nineteen forty-three is just found growing tips of the plant is very low, what have is even virus-free, if utilize method for tissue culture, getting a certain size shoot tip meristem cultivates, the plant of regeneration is not likely carried any virus, thereby can obtain virus-free seedling, then breeds with detoxic seedling, just can or seldom not there is not virus infections in the crop of plantation, can increase substantially ornamental value and the use value of crop.

Summary of the invention

For above-mentioned technical problem, the invention provides a kind of method for tissue culture of American red-maple, realize the artificial fast, economical of American red-maple and breed, improve the ornamental value of American red-maple, advance the further industrialization of American red-maple.

The present invention solves the problems of the technologies described above adopted technical scheme: a kind of method for tissue culture of American red-maple, comprises the following steps:

A: select the productive tiller section of American red-maple band axillalry bud as explant, first to the pretreatment that carries out disinfection of productive tiller section;

B: then on superclean bench, explant is inoculated in axillary bud development startup medium and is cultivated, obtain aseptic seedling;

C: above-mentioned aseptic seedling is inoculated in bud propagation and elongation medium and is cultivated;

D: select the growing point of propagation bud to transfer to successively in callus inducing medium, callus proliferated culture medium, root induction medium and cultivate according to cultivation progress, obtain nontoxic plant;

E: by nontoxic plantlet of transplant plantation.

As preferably, transplanting before plantation, nontoxic plant is carried out to hardening exercise.

As preferably, explant is inoculated into axillary bud development and starts in medium and carry out dark place reason 2d, and then proceed under 1600-2000Lux light intensity and cultivate, illumination 12-14h/d, temperature 22-26 ℃, cultivates 20 days, acquisition aseptic seedling.

As preferably, in step (c), condition of culture is: temperature 24-26 ℃, and humidity 60%-70%, illumination is 12-14h/d, under the fluorescent lamp that light intensity is 1600-2000Lux, cultivates.

As preferably, described hardening is taken exercise to adopt and is closed the mode of carrying out uncork hardening 3d after bottle hardening 6d and carry out.

As preferably, described American red-maple axillary bud development starts medium and is combined as MS+6-BA (0.6mg/L)+IAA (0.4mg/L).

As preferably, described bud propagation and elongation medium are combined as 1/2MS+6-BA (1.0mg/L)+IAA (0.2mg/L)+GA3 (1.0mg/L)

As preferably, described callus inducing medium is combined as WAP, 6-BA (0.6mg/L), IAA (0.3mg/L), callus proliferated culture medium is combined as WAP, 6-BA (0.6mg/L), IAA (1.0mg/L), and root induction culture medium prescription is 1/2MS, 6-BA (0.5mg/L), NAA (2.0mg/L), active carbon (1.0mg/L).

As can be known from the above technical solutions, the present invention adopts unique explant and culture medium prescription, has increased substantially success rate and plant quality that American red-maple tissue is cultivated, for the industrialized development of American red-maple provides strong guarantee, and cultivates quick, economical.

Embodiment

Introduce in detail cultural method of the present invention below, first select the productive tiller section of American red-maple band axillalry bud to cultivate as explant, first to productive tiller section, the pretreatment that carries out disinfection is inoculated into best axillary bud development explant and starts in medium on superclean bench, dark place reason 2d, then proceed under 1600-2000Lux light intensity and cultivate, illumination 12-14h/d, temperature 22-26 ℃, cultivate 20 days, obtain aseptic seedling, then aseptic seedling is inoculated in bud propagation and elongation medium, condition: temperature 24-26 ℃, humidity 60%-70%, illumination is 12-14h/d, light intensity is to cultivate under the fluorescent lamp of 1600-2000Lux, finally select the growing point of propagation bud to transfer to successively callus inducing medium according to cultivation progress, callus proliferated culture medium, in root induction medium, cultivate, the nontoxic plant of acquisition after cultivation completes, and then by nontoxic transplantation of seedlings plantation, in order to improve transplanting survival rate, employing is closed after bottle hardening 6d the mode of uncork hardening 3d and is transplanted front hardening and take exercise.

In implementation process, American red-maple axillary bud development starts cultivation optimal medium and is combined as MS+6-BA (0.6mg/L)+IAA (0.4mg/L), bud propagation and elongation medium are combined as 1/2MS+6-BA (1.0mg/L)+IAA (0.2mg/L)+GA3 (1.0mg/L), described callus induction optimal medium is combined as WAP+6-BA (0.6mg/L)+IAA (0.3mg/L), callus propagation is cultivated optimal medium and is combined as WAP+6-BA (0.6mg/L)+IAA (1.0mg/L), root induction optimal medium formula is 1/2MS+6-BA (0.5mg/L)+NAA (2.0mg/L)+active carbon (1.0mg/L).

Embodiment

The effective stem apex 1.0-1.5cm that gets American red-maple band axillalry bud cultivates explant as tissue, explant is put into the 20-30S that sterilizes in 70% alcohol on superclean bench, then proceed to the 5-10min that sterilizes in 2% liquor natrii hypochloritis, use again 5 left and right of aseptic water washing, handle and rear explant is inoculated into and in MS+6-BA (0.6mg/L)+IAA (0.4mg/L) medium, cultivates 15d left and right, obtain aseptic seedling, again aseptic seedling is inoculated in 1/2MS+6-BA (1.0mg/L)+IAA (0.2mg/L)+GA3 (1.0mg/L) medium after 20 days, obtain the bud of a large amount of propagation, finally on superclean bench, the growing point of these buds of picking is inoculated into WAP+6-BA (0.6mg/L)+IAA (0.3mg/L) medium culture about 10 days successively, WAP+6-BA (0.6mg/L)+IAA (1.0mg/L) medium culture is cultivated about 30 days and in 1/2MS+6-BA (0.5mg/L)+NAA (2.0mg/L)+active carbon (1.0mg/L) medium in surrounding left and right and is cultivated, thereby obtain tissue and cultivate nontoxic plant, if group training seedling is directly transplanted, survival rate is not high, in order to improve survival rate, first will transplant front hardening to group training seedling and take exercise, and general employing is closed after bottle hardening 6d+ uncork hardening 3d, then is transplanted in booth and goes.

Above-mentioned embodiment is used for illustrative purposes only, and be not limitation of the present invention, the those of ordinary skill in relevant technologies field, without departing from the spirit and scope of the present invention, can also make various variations and modification, therefore all technical schemes that are equal to also should belong to category of the present invention.

Claims

1. a method for tissue culture for American red-maple, comprises the following steps:

E: by nontoxic plantlet of transplant plantation.

2. method according to claim 1, is characterized in that: transplanting before plantation, nontoxic plant is carried out to hardening exercise.

3. method according to claim 1, is characterized in that: explant is inoculated in axillary bud development startup medium and carries out dark place reason 2d, then proceeds under 1600-2000Lux light intensity and cultivates, illumination 12-14h/d, temperature 22-26 ℃, cultivates 20 days, obtains aseptic seedling.

4. method according to claim 1, is characterized in that: in step (c), condition of culture is: temperature 24-26 ℃, and humidity 60%-70%, illumination is 12-14h/d, under the fluorescent lamp that light intensity is 1600-2000Lux, cultivates.

5. method according to claim 2, is characterized in that: described hardening is taken exercise to adopt and closed the mode of carrying out uncork hardening 3d after bottle hardening 6d and carry out.

6. method according to claim 1, is characterized in that: described American red-maple axillary bud development starts medium and is combined as MS+6-BA (0.6mg/L)+IAA (0.4mg/L).

7. method according to claim 1, is characterized in that: described bud propagation and elongation medium are combined as 1/2MS+6-BA (1.0mg/L)+IAA (0.2mg/L)+GA3 (1.0mg/L).

8. method according to claim 1, it is characterized in that: described callus inducing medium is combined as WAP, 6-BA (0.6mg/L), IAA (0.3mg/L), callus proliferated culture medium is combined as WAP, 6-BA (0.6mg/L), IAA (1.0mg/L), and root induction culture medium prescription is 1/2MS, 6-BA (0.5mg/L), NAA (2.0mg/L), active carbon (1.0mg/L).