CN102283127A - Culture medium synchronously used for induction and multiplication of Cymbidium hybridum stem tip Caespitose shoots and use method thereof - Google Patents

Abstract

The invention discloses a culture medium synchronously used for induction and multiplication of Cymbidium hybridum stem tip Caespitose shoots and a use method thereof. The culture medium comprises the following components by weight with a pH value of 5.8: content of major elements in an MS basic culture medium being halved, namely 1 L; 2 mg/L of 6- benzyl-purine; 0.3 mg/L of naphthyl acetic acid, 20 mg/L of sucrose and 8 g/L of agaragar. The use method of the culture medium comprises the following steps of; (1) shearing Cymbidium hybridum stem tip under an anatomical lens, inoculating on the induced and multiplied culture medium, and inducing Caespitose shoots; (2) inoculating the induced Caespitose shoots on the induced and multiplied culture medium to continuously multiply and propagate; (3) cutting the gemmae of the induced Caespitose shoots, and inoculating on the rooting culture medium for rooting; and (4) hardening off the rooted plantlets and transplanting. The culture medium used for inducing Cymbidium hybridum Caespitose shoots has high efficiency and low mutation rate, and is favorable for keeping good quality of the seedlings. With the adoption of the method to culture the Cymbidium hybridum, no protocorm stage is required from inducing the Caespitose shoots to rooting culture, and only two culture media are used for realizing culturing of normal four stages.

Description

A kind of medium and using method thereof that is used for hybrid cymbidium stem apex inducing clumping bud and propagation simultaneously

Technical field

The present invention relates to a kind of plant culture, belong to the hybrid cymbidium tissue culture technique, particularly a kind of medium and using method thereof that is used for hybrid cymbidium stem apex inducing clumping bud and propagation simultaneously.

Background technology

Hybrid cymbidium ( Cymbidium glandiflorium), be grow nonparasitically upon another plant kind and crossbreed thereof of a part of Da Hua in the orchid family Cymbidium (Cymbdiium).Its pattern is gorgeous, attitude is graceful, the florescence is long, is one of indoor pot flowers popular on present China flowers market.Hybrid cymbidium is the same with most of orchids, and the embryonic development of seed is incomplete, does not almost have endosperm, and the natural germination rate of seed is extremely low, procreation difficulty under nature, and traditionally by division propagation, the plant division ability is weak, the rate of increase is extremely low, reproduction speed is slow.Because long-term vegetative propagation, viruliferous plant is increasing, and quality descends, and causes the present commercially available seedling of China mainly to rely on external import.Adopting tissue culture technology to produce high-quality test-tube plantlet is an effective way that satisfies seedling market.

In the fast numerous prior art of hybrid cymbidium, to the cultured in vitro of hybrid cymbidium mainly by inducing shoot apical meristem to form protocorm or callus breaks up the acquisition test-tube plantlet again, though the method is induced the efficient height, lopsided seedling rate height is unfavorable for industrialization large-scale production.Publication number is a CN1586166(hybrid cymbidium high quality seedling method for quickly breeding), the cultural method of a CN101455178(new varieties hybrid cymbidium) and the CN1247022(quickly in-vitro breeding method of macranthus Hui orchid) three kinds of patents of invention all are to adopt the protocorm method to carry out the hybrid cymbidium tissue rapid propagation.Though (bud of growing thickly---hybrid cymbidium is the new way [J] of breeding fast for Liu Lifeng etc.; Sichuan Agricultural University's journal, 2004(2)) method of report is induced by a shoot apical meristem and is formed the bud of growing thickly, but bud of most stem apex differentiation induces efficient low, and stem apex rate of increase after cultivating 80 days just no longer changes.Adopting hybrid cymbidium stem apex or seed is in the conventional breeding of explant, is inducing, subculture, is growing seedlings and the four-stage of taking root uses different medium, and production procedure is many, the production cost height.

Summary of the invention

Purpose of the present invention, be at the deficiencies in the prior art, a kind of medium that can improve the inducing clumping bud rate and can be used to breed simultaneously is provided, can realize the conventional culture effect that needs four kinds of medium four-stages by this medium and root media, the seedling quality better of producing, simplify production routine, raise the efficiency, saved cost.

Such realization of the present invention, described a kind of medium that is used for hybrid cymbidium stem apex inducing clumping bud and propagation simultaneously is characterized in that: the amounts of components of this medium is,

Macroelement content reduces by half in the minimal medium, 1L;

The 6-benzyl purine, 2mg/L;

Methyl, 0.3mg/L;

Sucrose, 20mg/L;

Agar, 8g/L;

Ph5.8。

A kind of using method that is used for the medium of hybrid cymbidium stem apex inducing clumping bud and propagation simultaneously of the present invention is characterized in that:

1) under anatomical lens, strip the hybrid cymbidium stem apex, comprise shoot apical meristem, also comprise the leaf primordium part, can be greatly to having a slice lobus cardiacus, be seeded in above-mentioned induce with proliferated culture medium on, induce the bud of growing thickly;

2) bud of growing thickly that will induce be seeded in above-mentioned induce with proliferated culture medium on continue propagation and expand numerous;

3) the bud budlet that grows thickly that will induce is downcut, and is seeded on the root media (1/2MS+NAA 0.3mg/L) and takes root;

4) will take root and transplant after the seedling hardening.

Tissue cultivating seedling of the present invention is induced and condition of culture comprises temperature, illumination, and above-mentioned steps 1 to 4 cultivation temperature is 24 ± 2 ℃, illumination 1400lx; The used bud budlet that grows thickly of step 3, the above bud of its growth 2cm promptly can be used for culture of rootage and induces.

In the said components, macroelement content reduces by half in the minimal medium, provide plant tissue growth to grow required inorganic nutritive element and part organic nutrition element, 6-benzyl purine and methyl are plant growth regulator, have and promote cultured tissue to form the effect of the bud of growing thickly, sucrose provides tissue growth used carbon nutrition, and the main effect of agar is to solidify medium, plays a supportive role.The said components stirring and dissolving is even, and pH is adjusted into about 5.8 and gets final product.

The invention has the beneficial effects as follows that it is numerous soon that employing the present invention carries out hybrid cymbidium, lower to the size requirements that strips the hybrid cymbidium stem apex, what strip is not shoot apical meristem, comprises that also leaf primordium partly, can reduce operation easier greatly to having a slice lobus cardiacus; Adopt basal culture medium to grow thickly inducing of bud and breeding of hybrid cymbidium stem apex, can be when will carrying out root induction greater than the seedling about 2cm, other less buds are continued on this medium further enrichment culture; Use hybrid cymbidium stem apex of basal culture medium can induce the bud of growing thickly more than 20 at most at 80 days, induce the efficient height, but also can on this medium, continue propagation; This test utilizes the bud approach of growing thickly, cultivate culture of rootage from startup, do not need through the protocorm stage, only used two kinds of medium just to realize the cultivation of four-stage, simplify production routine, saved cost, improved efficient, reduced because the aberration rate that the process protocorm induces differentiation to cause ensures the seedling quality.

Embodiment

A kind of hybrid cymbidium stem apex inducing clumping bud and proliferated culture medium and using method thereof of being used for of the present invention, this medium is that 6-benzyl purine 6-BA, methyl NAA, sucrose and agar are dissolved in and will form (claiming 1/2MS) in the medium that macroelement content reduces by half among the minimal medium MS, and the consumption of its component is:

1/2MS (MS macroelement content reduces by half) 1L,

6－BA，2mg/L；

NAA，0.3mg/L；

Sucrose 20mg/L

Agar 8g/L

pH5.8

In the mentioned component, 1/2MS is that macroelement content reduces by half among the minimal medium MS, provide plant tissue growth to grow required inorganic nutritive element and part organic nutrition element, 6-BA and NAA are plant growth regulator, have and promote cultured tissue to form the effect of the bud of growing thickly, sucrose provides tissue growth used carbon nutrition, and the main effect of agar is to solidify medium, plays a supportive role.The said components stirring and dissolving is even, and pH is adjusted into about 5.8 and gets final product.

The present invention is used for the breeding of hybrid cymbidium, it adopts following steps: step 1: (the stem apex size is not a meristematic tissue to strip the hybrid cymbidium stem apex under anatomical lens, also comprise the leaf primordium part, can be greatly to having a slice lobus cardiacus), be seeded in of the present invention induce with proliferated culture medium on, induce the bud of growing thickly; Step 2: the bud of growing thickly that will induce is seeded in of the present invention inducing and continues to expand numerous with proliferated culture medium; Step 3: the bud budlet that grows thickly that will induce is downcut, and is seeded on the root media (1/2MS+NAA 0.3mg/L) and takes root; Step 4: transplant after the seedling hardening of will taking root.Tissue cultivating seedling is induced and condition of culture comprises temperature, illumination, and step 1 to four, cultivation temperature condition are 24 ± 2 ℃, illumination 1400lx.The used above bud of bud budlet its growth 2cm of growing thickly of step 3 promptly can be used for culture of rootage and induces.So only with two kinds of medium (1, induce and proliferated culture medium; 2, root media), just realize the cultivation of four-stage, simplified production routine, saved cost, improved efficient,

Can make the hybrid cymbidium stem apex induce the bud of growing thickly with the present invention, induce efficient to be not less than 90%, one stem apex and can induce more than 20 bud at most in 80 days, significantly improve reproduction coefficient, be easy to batch production and produce the hybrid cymbidium tissue cultivating seedling in cultivation.

Claims (2)

1. medium that is used for hybrid cymbidium stem apex inducing clumping bud and propagation simultaneously is characterized in that: the component consumption of this medium is,

Macroelement content reduces by half in the minimal medium, 1L;

The 6-benzyl purine, 2mg/L;

Methyl, 0.3mg/L;

Sucrose, 20mg/L;

Agar, 8g/L;

Ph5.8。

2. using method that is used for the medium of hybrid cymbidium stem apex inducing clumping bud and propagation simultaneously is characterized in that:

1) under anatomical lens, strip the hybrid cymbidium stem apex, comprise shoot apical meristem, also comprise the leaf primordium part, can be greatly to having a slice lobus cardiacus, be seeded in above-mentioned induce with proliferated culture medium on, induce the bud of growing thickly;

2) bud of growing thickly that will induce be seeded in above-mentioned induce with proliferated culture medium on continue propagation and expand numerous;

3) the bud budlet that grows thickly that will induce is downcut, and is seeded on the root media and takes root;

4) will take root and transplant after the seedling hardening.