JPH0956286A - Mass propagation of genus gmelina tree using tissue culture - Google Patents

Abstract

PROBLEM TO BE SOLVED: To propagate a tree belonging to genus Gmelina useful for a building material, etc., in a large amount for a short period of time by culturing a shoot apex of trees belonging to genus Gmelina, inducing and propagating a multibuds body having many (in)definite buds and regenerating a plant body under a specific condition. SOLUTION: A shoot apex of a tree belonging to genus Gmelina such as Gmelina arborea Roxb is cultured, a multibuds body having many (in)definite buds is induced and propagated, and a large amount of shoot is produced from the multibuds body, then the shoot is transplanted into a rooting medium such as a B5 medium containing 0.002mg/L of indolebutyric acid (IBA) with non- adding of a growth-regulating substance of plants, or together with 0.01-0.3mg/V of benzine aluminopurine (BAP), thus rooted and a plant body is regenerated, then a tree belonging to genus Gmelina is propagated in a large amount. Preferably, a B5 medium containing 0.01-0.3mg/L BAP or 0.01-0.3mg/L BAP with 0.002mg/L IBA, or its modified medium is used as a medium producing the shoot.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a tree belonging to the genus Gmelina, particularly Gmelina belonging to the genus Gmelina.
Arborea R _OXB .

[0002]

2. Description of the Related Art Gmelina trees are major afforested trees in tropical regions of the world (India, Southeast Asia),
The materials are building materials, engraving materials, pulp, fiberboard,
The bark, roots and fruits of plywood are fast-growing trees used for medicinal purposes. In recent years, due to the development of tissue culture technology, preservation of excellent varieties using tissue culture has been performed on many trees for the purpose of preservation of genetic resources, and even for Gmelina trees, tissue culture is performed for preservation of excellent varieties. The use of technology is expected. In addition, generally, when trying to propagate useful old-aged trees, existing methods such as cuttings cannot be used,
Propagation of elite trees is considered difficult. In recent years, several research groups have conducted research on tissue culture technology in Gmelina trees for the purpose of solving these problems, but only two successful cases have been obtained to date (JC Yang. , G.Y.Tsay, J.D.Ch.
ung, Z. Z. Chen (1992) Microp
ropagation of Gmelina arbo
rea R. , Proceedings of the
SABRAO international symp
osium 29: 213-218, and Crisald.
o, E. N. (1980) Tissue culture
e of fast-growing trees, S
ylvatrop 5 (2); 123-137). However, these methods use seedlings germinated under aseptic conditions as materials, and achieve regeneration and mass growth of seedlings from materials collected from adult trees containing a large amount of phenolic substances. I can't. Therefore, development of an efficient culture method from mature trees is desired.

[0003]

The object of the present invention is to provide Gm
Trees of the genus erina, in particular Gmelina arbore
a _Providing a mass propagation method that enables mass production of cloned seedlings of R _OXB .

[0004]

The present inventor has found that Gmeli
As a result of diligent research for the purpose of developing a mass-propagation method capable of mass-production using saplings of Na genus trees or adult trees, as a result, the shoot apex of Gmelina genus tree was cultivated, The present invention succeeded in inducing a multiblast having an adventitious bud and efficiently propagating the multiblast. In particular, by adding benzylaminopurine (BAP) or indoleacetic acid (IBA) and BAP to the medium used for inducing multiblasts and the medium used for proliferating multiblasts, it is possible to efficiently produce multiple buds. It was found that the body can be induced and proliferated, and shoot elongation can be efficiently promoted from the proliferated polyblasts. The present inventors have found that further growth and rooting of shoots can be achieved by culturing the proliferated shoots in a medium containing no plant growth regulator, or a medium containing BAP or BAP and IBA, thus completing the present invention.
That is, the present invention cultivates the shoot apex of a tree of the genus Gmelina to induce a multibud body having a large number of regular buds and / or adventitious buds,
This is a method for mass-growing Gmelina genus trees, which comprises proliferating and producing a large amount of shoots from the multibuds, and then transplanting the shoots into a rooting medium for rooting to regenerate the plant.

According to the present invention, by inducing and proliferating a multibud body from the shoot apex of a tree of the genus Gmelina and grown on the shoot apex, and then extending shoots from the resulting multibud body and rooting the shoot. Therefore, a large amount of seedlings can be efficiently produced. More specifically, the genus Gmelina targeted by the present invention is Gmelina arbor.
ea R _OXB . As the culture material used in the present invention, shoot apex collected from a sapling or an adult tree of a tree of the genus Gmelina is used. The collected shoot apex is surface sterilized with ethanol and sodium hypochlorite, hydrogen peroxide water, or mercury chloride (elevation water) according to a usual method, washed with sterilized water, and then cultured in a medium. As a basal medium used for culture, a medium containing an inorganic component and a carbon source as essential components and other plant growth regulators, vitamins and amino acids is used. As the inorganic component, an inorganic compound containing an element such as nitrogen, phosphorus, potassium, sodium, calcium, sulfur, iron, manganese, zinc, iodine, boron, molybdenum, chlorine or cobalt is used. Carbohydrates such as sucrose or glucose are used as carbon sources. Auxin and cytokinin are used as plant growth regulators. Examples of auxins include 3-indole butyric acid (IBA) and naphthalene acetic acid (NAA), and examples of cytokinins include benzylaminopurine (BAP), kinetin, zeatin, and 4-phenylurea (4PU). . Examples of vitamins include thiamine, pyridoxine, nicotinic acid and the like. As an amino acid,
Examples thereof include glycine, glutamic acid, lysine and the like.

The medium used for actual culture is a medium used for plant tissue culture, such as MS medium (Murashige, T. (1962), Physi).
ol. Plant. 15: 473-497), B5 medium (Gamborg, OL (1968), Exp. C).
ell. Res. 50: 151-158), WP medium (Lloyd, G. (1981), Int. Plant.
Prop. Soc. 30: 421-427), BTM
Medium (Challupa, V. (1984) Biolog
ia Plnt. (Praha) 26: 374-37
7) etc. are mentioned. Particularly, B5 medium and its modified medium are preferable. In order to promote the induction of fixed buds, adventitious buds and their growth, and shoot elongation from multiple shoots, BAP was added to 0.
01-0.3 mg / l, or BAP 0.01-0.
It is preferable to use a B5 medium containing 3 mg / l and about 0.002 mg / l of IBA or a modified medium thereof.

Then, the elongated shoots are divided or transplanted into a rooting medium without division, whereby the shoots can be further elongated and rooted. The rooting medium contains the above-mentioned inorganic components, carbon source, vitamins, amino acids, etc., and contains no plant growth regulator, or BAP 0.01-0.3 mg / l and IBA 0.002 m.
A medium containing about g / l is preferable.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION Gmel by the tissue culture of the present invention
As one of the preferred embodiments of the method for mass-growing the ina genus tree, first, Gmelina arborea R is used.
_The shoot apex is collected from the adult _OXB tree, and IBA is 0.002 mg.
/ L and BAP were cultured in a modified B5 medium containing 0.01 to 0.3 mg / l to induce and proliferate multispores having a large number of regular buds and / or adventitious buds, and further shoot a large amount of shoots from the multispores. To produce. Then, the obtained shoot is
The plant can be efficiently regenerated by transplanting into a rooting medium which is a modified B5 medium containing 0.01 to 0.3 mg / l and IBA 0.02 mg / l and being rooted.

[0009]

INDUSTRIAL APPLICABILITY According to the present invention, Gmel can be obtained from materials collected from saplings or adult trees of the genus Gmelina.
Aseptic seedlings, seedlings and cuttings of ina trees can be mass-produced in vitro in a short period of time, and Gmelin
The possibility of mass production of seedlings of a-genus tree increases.

[0010]

The present invention will be described in more detail with reference to the following examples. Gmelina arborea R _OXB. Seedlings (height _0.2-0.5 m) and mature trees (6.0-10.0) _.
Material was taken from m). (1) Sterilization of material The top buds were collected from the seedlings and mature trees of Gmelina arborea R _OXB.
% Surface sterilization was carried out in sodium hypochlorite for 6 minutes, washed several times with sterilized water, and then air-dried on sterilized filter paper. After air-drying, the outer skin of the apical bud was removed under an optical microscope, the shoot apex was extracted, and cultured on an agar medium. As a result, seedlings, the survival rate of the material collected from the adult was slightly better than the material collected from the seedling, according to the method of the present invention, both seedlings, or the material collected from the adult Could be used. Table 1
The data showing the comparison of the survival rate of the shoot apices collected from the seedlings and the adult trees is shown in FIG.

[0011]

[Table 1] Survival rate of collected material. ━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━━ Material Survival rate (%) Mortality rate (%) Contamination rate (%) ) ────────────────────────────────── Stem apex (from mature tree) 70 10 20 Stem apex (from seedling) ) 86 10 4 ──────────────────────────────────

(2) Induction, proliferation and shoot elongation of multiblasts As a medium, modified B5 medium (B5 medium in which all medium components are halved), sucrose 20 g / l and gellite 3.4 g are used.
/ L and 0.002m of IBA as a plant growth regulator
g / l, 0.01 to 0.3 mg / l of BAP, p
After adjusting H to 5.7, it was sterilized before use. Culture temperature is 26
± 2 ℃, 16 hours a day fluorescent lamp (3,000 ~
(5,000 lux) illumination. Multiblasts were induced after 2-3 months of culture. In particular, it was effective to add 0.002 mg / l of IBA and 0.03 mg / l of BAP, and no significant difference was observed depending on the material (Fig. 1,
3). (3) Rooting The large number of shoots obtained in (2) were cut into individual shoots and transplanted into a rooting medium. As the medium, a modified B5 medium containing no plant growth regulator, or BAP0.01-
A modified B5 medium (same as the above (2)) containing 0.3 mg / l and 0.002 mg / l of IBA was adjusted to pH 5.7 and sterilized before use. Culture temperature is 26 ± 2 ° C., 1
16 hours per day fluorescent lamp (3,000-5,000 lu
x) Illumination. As a result, after 1-2 months of culture, 3
Rooting was observed in 3.3% of individuals, and 3-10 cm at the same time
Shoots can be extended and 2 to 8 new leaves can be developed. Especially, a medium without addition of a plant growth regulator, or BAP 0.01-0.03 mg / lmg / l and IB
It is effective to add A 0.002 mg / l,
No significant difference was observed depending on the material (Figs. 2 and 4).

[Brief description of drawings]

FIG. 1 is a graph showing the influence of IBA and BAP concentrations on the induction of multiblasts when seedling-derived material was used.

[Fig. 2] I for rooting when materials derived from seedlings were used
It is a graph which shows the influence of BA and BAP.

FIG. 3 is a graph showing the influence of IBA and BAP concentrations on the induction of multiblasts when a material derived from mature wood is used.

[FIG. 4] I for rooting when a material derived from an adult tree is used
It is a graph which shows the influence of BA and BAP.

Claims (5)

[Claims] 1. Incubating the shoot apex of a tree of the genus Gmelina to induce a multibud body having many fixed buds and / or adventitious buds,
A method for mass-growing Gmelina genus trees, which comprises proliferating and producing a large amount of shoots from a multibud, and then transplanting the shoots into a rooting medium to cause rooting to regenerate a plant. 2. Incubating the shoot apex of a tree of the genus Gmelina to induce a multibud body having a large number of fixed buds and / or adventitious buds,
Benzylaminopurine (BAP) was added as a medium to grow and further produce a large amount of shoots from multiblasts.
01-0.3 mg / l, or BAP 0.01-0.
Indole butyric acid (IBA) 0.0 with 3 mg / l
The mass proliferation method according to claim 1, wherein B5 medium containing 02 mg / l or a modified medium thereof is used. 3. As a rooting medium, a plant growth regulator-free B5 medium containing 0.002 mg / l of IBA together with 0.01 to 0.3 mg / l of BAP, or a modified medium thereof is used. Alternatively, the mass proliferation method described in 2. 4. The mass propagation method according to claim 1, wherein a shoot apex from a sapling or an adult tree of Gmelina genus tree is used. 5. The tree of the genus Gmelina is Gmelina.
Arborea R _OXB. 5. The large-scale proliferation method according to any one of claims 1 to 4.