CN104885932A

CN104885932A - Tissue culture and rapid propagation method for rhododendron moulmainense

Info

Publication number: CN104885932A
Application number: CN201410795987.0A
Authority: CN
Inventors: 庄雪影; 孙朝辉; 赵富群; 洪文君
Original assignee: South China Agricultural University
Current assignee: South China Agricultural University
Priority date: 2014-12-11
Filing date: 2014-12-11
Publication date: 2015-09-09

Abstract

The invention provides a tissue culture and rapid propagation method which is most suitable for rhododendron moulmainense. Complete plants are obtained through the steps of obtaining bacteria-free seedlings, obtaining explants, inducing and differentiating adventitious buds, performing propagation culture and performing rooting culture sequentially; and by hardening seedlings and transplanting the seedlings, the survival rate of nursery stocks is increased. The tissue culture and rapid propagation method disclosed by the invention has the beneficial effects that (1) a tissue culture and rapid propagation system for the rhododendron moulmainense is established, and a tissue culture technique gap of the rhododendron moulmainense in the prior art is filled; (2) the optimum propagation culture medium Read+1.0mg/L ZT+0.02mg/L IBA+2.5%-3.5% cane sugar is screened, and the rooting culture medium is 1/2 WPM+1.5mg/L IBA; and (3) a suitable seedling hardening and transplanting method of the rhododendron moulmainense is researched.

Description

The tissue culture and rapid propagation method of a kind of velvet apple cuckoo

Technical field

The present invention relates to a kind of flowers propagation technique field, be specifically related to tissue cultures, the method for quickly breeding of a kind of velvet apple cuckoo.

Background technology

Velvet apple cuckoo Rhododendron moulmainense is the evergreen dungarunga of Ericaceae, natural distribution in China, India, Indonesia, Malaysia and Burma, in the forest being mainly distributed in height above sea level 400-1500m on the south the Yangtze river basin in China and shrubbery.Velvet apple cuckoo flower is gorgeous, young leaves is red tender, is the fine tree species building flowering shrubs and Hua Qun view.But its seedling poor growth under normal conditions, seedling variation is large.Have been reported though numerous research is expanded in cuttage, it expands numerous amount by maternal plant material and the impact of mating season, and is difficult to the requirement reaching scale Fast-propagation.Tissue cultures raising technology has that reproduction coefficient is high, reproduction speed soon, not by seasonal effect be easy to the advantages such as industrialization, in order to protect, develop and useining velvet apple azalea resource better, the tissue culture technique of research velvet apple cuckoo is significant.

Therefore, the application, for the key link of velvet apple cuckoo group culturation rapid propagating technology, has carried out the key technology screening studies such as minimal medium, sucrose, growth hormone, the basic element of cell division, hardening, has established a set of velvet apple cuckoo Plant Tissue Breeding system.

Summary of the invention

The object of the invention is to: provide a kind of cultivation cycle short, by the tissue culture and rapid propagation method of seasonal effect, velvet apple cuckoo that output is higher, to overcome the deficiencies in the prior art, promote the large-scale application of velvet apple cuckoo.

Technical scheme of the present invention is: the tissue culture and rapid propagation method of a kind of velvet apple cuckoo, is characterized in that: comprise the steps:

A. the acquisition of aseptic seedling: choose full seed and wrap in gauze, 2h is rinsed under being first placed in running water, with 75% alcohol-pickled 2min, aseptic water washing 5 times, add tween 2 sterilization 90s with 0.1% mercuric chloride again, aseptic water washing 5 times, blots the residual moisture of the surface of the seed with sterilizing filter paper, be inoculated in and only with the addition of 0.6% agar, 25% sucrose, pH is in the minimal medium of 5.8.

B. Multiplying culture: explant Multiple Buds is inoculated in cultivate in the Read medium proliferated culture medium that is minimal medium, every 60d subculture once.

C. culture of rootage: the healthy and strong seedling segment obtained by Multiplying culture, is inoculated in cultivate root induction in the 1/2WPM medium root media that is minimal medium.

Preferably, described proliferated culture medium is Read+1.0mg/L ZT+0.02mg/L IBA+2.5%-3.5% sucrose, and its proliferation times reaches 4.5.

Preferably, described root media is 1/2WPM+1.5mg/L IBA, and its rooting rate can reach 80.17%; Adding active carbon 0.1% can hestening rooting number and the long-living length of root.。

In addition, also comprise hardening and transplant step, concrete grammar is: after culture of rootage 15d, after cultivating 3-5d, is unscrewed by blake bottle lid, transplants seedlings after 7d under blake bottle being moved to room temperature environment, natural lighting environment; Plant root medium is washed away under flowing water, clean plant root is soaked in 2-2.5h in certain density medicament, with peat soil and perlite volume ratio 3: 1 for transplanting medium, transplant in the cave dish in 32 caves, in front 10d after transplanting, hide film moisturizing, control relative moisture 75%-85%, after 10d, remove film gradually.Described medicament is: 5-ALA solution, and its concentration is 5ppm.

After removing film 15d, pouring concentration is the solution of the 5-ALA of 50ppm, after this, repeats pouring once, transplant seedlings until change basin next time every 15d.

Accompanying drawing explanation

Fig. 1 is Read medium treatment group.

Fig. 2 is hardening processed group.

Fig. 3 is not for add archusia processed group.

Fig. 4 is for adding archusia processed group.

Fig. 5 is processed group of taking root.

Fig. 6 is charcoal treatment group.

Embodiment

Below in conjunction with accompanying drawing, the invention will be further described in fact.

A. the acquisition of aseptic seedling: choose full seed and wrap in gauze, 2h is rinsed under being first placed in running water, with 75% alcohol-pickled 2min, aseptic water washing 5 times, add tween 2 sterilization 90s with 0.1% mercuric chloride again, aseptic water washing 5 times, blots the residual moisture of the surface of the seed with sterilizing filter paper, seed after sterilizing is inoculated in and adds 1/4MS, 0.6% agar, 25% sucrose, in the minimal medium of pH 5.8.

B. Multiplying culture: be inoculated in by explant Multiple Buds in the proliferated culture medium of Read+1.0mg/L ZT+0.02mg/L IBA+2.5%-3.5% sucrose, once, proliferation times reaches 4.5 to every 60d subculture.

C. culture of rootage: the healthy and strong seedling segment obtained by Multiplying culture, be inoculated in the root media into 1/2WPM+1.5mg/L IBA, its rooting rate can reach 80.17%, and adding active carbon 0.1% can hestening rooting number and the long-living length of root.

D. hardening and transplanting: after culture of rootage 15d, after cultivating 3-5d, unscrews blake bottle lid, unscrews, transplant seedlings after 7d under blake bottle being moved to room temperature environment, natural lighting environment; Plant root medium is washed away under flowing water, clean plant root is soaked in 2.5h in the 5-ALA solution that concentration is 5ppm, with peat soil and perlite volume ratio 3: 1 for transplanting medium, transplant in the cave dish in 32 caves, in front 10d after transplanting, hide film moisturizing, control relative moisture 80%, after 10d, remove film gradually.After removing film 15d, pouring concentration is the solution of the 5-ALA of 50ppm, after this, repeats pouring once, transplant seedlings until change basin next time every 15d.

This tissue culture technology system provides a set of applicable velvet apple cuckoo Fast-propagation, solves the problems such as velvet apple cuckoo seedling growth is slow, the breeding cycle is long, and growth of seedling state is poor, large-scale culture can go out healthy and strong nursery stock.

Up to now, to the research of the tissue culture technique of velvet apple cuckoo, there is not been reported, the reported first of the present invention technical system of applicable velvet apple cuckoo tissue-culturing quick-propagation.

Finally to should be noted that; above embodiment is only in order to illustrate technical scheme of the present invention but not limiting the scope of the invention; although be explained in detail the present invention with reference to preferred embodiment; those of ordinary skill in the art is to be understood that; can modify to technical scheme of the present invention or equivalent replacement, and not depart from essence and the scope of technical solution of the present invention.

Claims

1. a tissue culture and rapid propagation method for velvet apple cuckoo, is characterized in that: comprise the steps:

A. the acquisition of aseptic seedling: choose full seed and wrap in gauze, 2h is rinsed under being first placed in running water, with 75% alcohol-pickled 2min, aseptic water washing 5 times, add tween 2 sterilization 90s with 0.1% mercuric chloride again, aseptic water washing 5 times, blots the residual moisture of the surface of the seed with sterilizing filter paper, be inoculated in and only with the addition of 0.6% agar, 25% sucrose, pH is in the minimal medium of 5.8;

B. Multiplying culture: explant Multiple Buds is inoculated in cultivate in the Read medium proliferated culture medium that is minimal medium, every 60d subculture once;

2. the tissue culture and rapid propagation method of a kind of velvet apple cuckoo according to claim 1, is characterized in that: described proliferated culture medium is Read+1.0mg/LZT+0.02mg/L IBA+2.5%-3.5% sucrose, and its proliferation times reaches 4.5.

3. the tissue culture and rapid propagation method of a kind of velvet apple cuckoo according to claim 1, is characterized in that: described root media is 1/2WPM+IBA 1.5mg/L, and its rooting rate can reach 80.17%; Adding active carbon 0.1% can hestening rooting number and the long-living length of root.

4. the tissue culture and rapid propagation method of a kind of velvet apple cuckoo according to claim 1, it is characterized in that: also comprise hardening and transplant step, concrete grammar is: after culture of rootage 60d, after cultivating 3-5d under blake bottle being moved to room temperature environment, natural lighting environment, blake bottle lid is unscrewed, transplants seedlings after 7d; Wash away plant root medium under water, clean plant root is soaked in 2-2.5h in certain density medicament, with peat soil and perlite volume ratio 3: 1 for transplanting medium, transplant in the cave dish in 32 caves, in front 10d after transplanting, hide film moisturizing, control relative moisture 75%-85%, after 10d, remove film gradually.

5. the tissue culture and rapid propagation method of a kind of velvet apple cuckoo according to claim 4, is characterized in that: described medicament is: 5-ALA solution, and its concentration is 5ppm.

6. the tissue culture and rapid propagation method of a kind of velvet apple cuckoo according to claim 4, is characterized in that: after removing film 15d, and pouring concentration is the solution of the 5-ALA of 50ppm, after this, repeats pouring once, transplant seedlings until change basin next time every 15d.