JPH0257130A - Production of adventitious embryo and/or adventitious bud of jasmine plant - Google Patents

Abstract

PURPOSE:To accomplish a stationary and stabilized production of seedlings by using a culture medium containing cytokinin and auxin for the culture of jasmine plant pieces to efficiently produce in a short time the adventitious embryos and/or adventitious buds of jasmine plant through tissue culture. CONSTITUTION:A culture medium is incorporated with each 0.1-5ppm of auxin (e.g., indole acetate) and cytokinin (e.g., kinetin). Thence, the sterilized tissue of a jasmine plant is put to culture in said medium pref. at 20-30 deg.C, and the resultant callus is transferred to a second medium under the same conditions as those for the first medium to repeat its culture.

Description

DETAILED DESCRIPTION OF THE INVENTION FIELD OF INDUSTRIAL APPLICATION The present invention relates to an efficient method for propagating jasmine (Jasullinum) humus, which is used as a raw material for fragrances and as an ornamental tree, by tissue culture.

K Conventional technology Σ Jasulinum humic plant
nd) and others Oubai (Jasun+inuII)
Approximately 200 species of plants such as Nudiflorum liu (formerly known as nudiflorum liu) are distributed in the tropics to subtropics, and most of them are woody and bear white, yellow, red, or pink flowers.

Among these, several types of white flowers contain aromatic essential oils mainly composed of jasmon, so they have been prized as raw materials for fragrances and as ornamental trees since ancient times, and are cultivated for commercial purposes. However, with the improvement of cultural life in recent years, the demand for these seeds has increased rapidly, and a stable supply of seeds and seedlings is strongly desired. However, conventional vegetative propagation methods such as division and cuttings have extremely low propagation rates and efficiency. In addition, there are many other problems, such as not only a large number of parent plants are required, but also a lot of manpower, and the production cost of seedlings is high. On the other hand, in an effort to establish a method for propagating woody plants, we have recently begun growing poplars and elms.

Research using tissue culture methods has been conducted on forest trees such as eucalyptus and orange, [l1lIIllCulture J 13(9)
, 292-299 (1987), etc., but no research has been published regarding plants of the genus Jasmine.

K Problems to be Solved by the Invention The purpose of the present invention is to provide a method for propagating jasmine humic plants to efficiently produce seedlings of plants of the genus jasmine in a short period of time, and more specifically, to provide a method for propagating plants of the genus jasmine by tissue culture. The present invention provides a method for efficiently producing somatic embryos and/or somatic buds.

Means for Solving Problem K] As a result of research on a method for efficiently producing large quantities of somatic embryos and/or adventitious buds of plants of the genus Jasmine in a short period of time, the present inventors have found that tissue pieces of plants of the genus Jasmine are treated with auxin. They have perfected a production method in which callus is induced by culturing in the dark and forming somatic embryos and/or adventitious buds in the light after subculture. .

Specific examples of plants of the genus Jasmine of the present invention include Osmanthus (Olea Seae) H.
Jasun+inuIllsambac 5oland
), Jasuminum offi
cinale l-, var, grandifloru
m Baey), others, mountain jasmine (Ja
Sull in LIInhemsleyi yall
lallOtO), rose-colored jasmine (Jasu)
m+numbeesianum Forest&Di
els), Jasmine of the Golden Coast (Jasunlin)
ulIldichton+um Vahl), Jasuminunlfloridum
Bunge), Anless Jasminu
m fluminense Vellozo).

Jasuminum gracilli
nlun+ Hook, F, ), etc. In order to propagate jasmine humic plants by the lltllli@ cultivation method, it can be carried out by first inducing callus and then forming somatic embryos and/or adventitious buds in the callus. In order to induce callus from tissue pieces of plants of the genus Jasmine, the stem, eastern part, stalk,
Tissues such as the stem apex, corolla, roots, etc. are cut into pieces of about 1 to 6 III/Ill squares, and the tissue pieces are then processed into the commonly used,
For example, after sterilization using an ethanol aqueous solution, a chloric acid aqueous solution, sterilized water, etc., the plate is placed in a medium containing effective amounts of cytokinin and auxin as active ingredients, and cultured at a temperature of 15 to 35°C in the dark. Preferably 20
This can be done by culturing at ~30°C. The callus thus obtained is transferred to a medium under the same conditions and cultured repeatedly to perform subculturing and obtain stable callus with a fast growth rate.

The callus was cut into pieces of about 5 mg, placed in a medium containing effective amounts of cy-1-kainin and au-4-sin as active ingredients, and white light with a brightness of about 1,400 lux was applied to the medium at a temperature of 15 to 35. A large number of somatic embryos and/or adventitious buds are formed on the surface of the callus by culturing with illumination for 8 to 24 hours per day at 1°C, preferably 20 to 30°C.

The medium used in the present invention is based on various known inorganic synthetic media, to which sugars, plant growth regulators, vitamins, amino acids, natural substances, etc. are added, and a solidifying agent such as agar or gellan gum is added. It can be used as a solid medium or a liquid medium without adding a solidifying agent.

A typical synthetic medium is H8 medium (Hurashi
geand Skoog's medium), white (14h
ite), Linsmeyer and Skoog (Li
nsmaier and skoog) medium, Niche (
Usual basic media for plant tissue culture, such as Nitsch's medium, as well as those with modified compositions of these basic media, and the like.

The inorganic synthetic medium is known as Jil (organic ingredient, teriki compound).

Sugars, vitamins, and amino acids are blended, and natural substances such as casein hydrolyzate, coconut milk, yeast extract, malt extract, and polypeptone are blended.

Auxin and cytokinin are common plant growth regulators, and indole'@
acids, α-naphthaleneacetic acid, indolebutyric acid, 2,4-dichlorophenogynoacetic acid and derivatives thereof,
These play the role of increasing the permeability of cell membranes, promoting cell division, suppressing the growth of lateral buds, and promoting the formation of adventitious roots.
Its action is stimulatory at low concentrations and inhibitory at high concentrations. Cytokinins include kinetin, zeatin, benzyladenine, and their H4 forms, and these play roles such as promoting the growth of buds and leaf growth, and preventing aging.

Somatic embryos and/or callus derived from tissues of plants belonging to the genus Jasmine
Alternatively, to induce adventitious buds, auxin and cytokinin may be added individually, but in this case, induction takes a considerable number of days and the number of induced somatic embryos and/or adventitious buds is small. , resulting in poor efficiency and undesirable results. In order to efficiently form somatic embryos and/or adventitious buds in callus derived from tissues of jasmine humic plants, it is preferable to use a mixture of auxin and cytokinin within a range of 0.1 to 5 ppm.

The present invention will be described in detail below with reference to specific examples, but the present invention is not limited to this scope.

Example 1゜The stems of Matsurica were washed with water, and 3.5% of the surfactant Tween 80 (trade name: Tween 80, manufactured by Tokyo Kasei Kogyo @) was added.
After sterilizing for 10 minutes with sterilized water containing 0.5 d per 100 units of aqueous sodium chlorate solution, they were washed three times with sterile water. This stem was cut into 5 mm pieces and treated with pl+ containing 5 ppIll of α-naphthalene acetic acid and 1 ppm of benzyladenine. 5.7. The cells were placed on MS medium containing 0.7% agar and cultured at 25°C in the light. After 2 to 3 weeks, enlargement of the stem was observed, and after 5 weeks, a callus with a diameter of about 1 cm was obtained. The callus was transplanted into the above-mentioned medium having the same composition as the medium used to obtain the callus, and this culture was repeated five times to obtain a stable callus mass with a fast growth rate. The callus thus obtained was crushed into small pieces of about 5 mg, placed in a medium with the same composition as above, and cultured for 40 days at 25°C under white light illumination of 1,400 lux and repeated irradiation for 16 hours per day. As a result, formation of 7 to 8 adventitious buds was observed on the callus surface.

The adventitious buds were cut out and grown in a medium with the same composition as above for an additional 20 days.
When culture was continued under the same conditions for one day, the plant again grew into a plant with organs.

Example 2 The callus mass derived from the stem obtained in Example 1 was cut into small pieces of about 5 my and prepared with agar-containing white (pH 5, 7, 0, 7%) containing 5 ppm of indole acetic acid and 1 ppn of benzyladenine. 14hite) and cultured for 40 days at 25°C under white light illuminance of 1,400 lux for 16 hours a day.
Adventitious buds were formed. The callus that had formed the indeterminate odor was further cultured for two more times under the same conditions using a medium with the same composition as described above.
When the culture was continued for 0 days, the adventitious buds further elongated and grew into plants with regrowth organs.

Example 3゜The callus strain derived from the stem obtained in Example 1 was crushed into small pieces of 2 to 5 IRg lumps, and an H8 liquid containing 5 ppIll of indole acetic acid and 1 ppn of benzyladenine with a pH of 5.7 without the addition of a solidifying agent was prepared. The cells were transferred to a medium and cultured for 3'ig in the dark at 25°C while rotating the medium at a speed of 1100 rp for proliferation.

The calli were transferred to MS solid medium containing 0.7% agar at PL+5.7 containing 5 ppm indoleacetic acid and 1 ppm benzyladenine, and irradiated with white light at 1,400 lux for 16 hours a day at 25°C. The cells were cultured for 40 days.

Many adventitious buds were observed on the callus surface. When the adventitious buds were further cultured on the same medium under the same conditions for 20 days,
The adventitious buds further elongated and grew into plants with organs.

Effects of the invention Σ According to the present invention, somatic embryos and/or adventitious buds of plants of the genus Jasmine can be efficiently produced in a short time using callus of plants of the genus Jasmine that have been mass-cultured in an artificial environment as raw materials. Therefore, it is possible to constantly and stably stock a large amount of seeds and seedlings of plants belonging to the genus Di17, which is very useful industrially.

Patent Applicant: Fujibo Co., Ltd. Agent, Patent Attorney: Katsu Ohno, Patent Attorney: Rei Ohno, Representation of Child Case Showa Proceedings Supplemented Patent Application (Sponsorship) No. 206181 (1) Specification, Page 3, Line 19 Eye [(Jasun+inum
dichtomumVahl)J to ``(JaSuln
inLIII dichotoIIllull Vah
l) Correct to J.

(2) "Jasmine" on page 6, line 15 of the specification cylinder is corrected to "1 jasmine".

3. Person who makes corrections Relationship with the incident full name How to create adventitious buds

Claims (1)

[Claims] 1. A plant piece of the genus Jasmine is placed on a medium containing cytokinin and auxin, a callus is induced in the dark, and after subculturing, somatic embryos and/or adventitious buds are formed in the light. Somatic embryos of plants of the genus Jasmine and/
Or a method for producing adventitious buds.