JPH0322928A - Formation and proliferation of tuberous tissue of plant of genus asparagus and proliferation of seedling - Google Patents

Abstract

PURPOSE:To form a tuberous tissue of a plant of genus Asparagus capable of proliferating large amount of plant or seedling in high efficiency by culturing the tissue or callus of the plant. CONSTITUTION:The tissue (preferably crown) or callus of a plant of genus Asparagus (preferably edible asparagus) is cultured to form a tuberous tissue. Preferably, the tissue is sliced and the obtained slice is cultured to thicken the tuberous tissue. Furthermore, similar proliferation process comprising (tuberous tissue slicing and culture thickening of tuberous tissue) is repeated, as necessary.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to a method for forming and propagating tuber-like tissues of plants of the genus Asparagus, and a method for propagating plants and seedlings. [Prior Art] Plants of the genus Asparagus have traditionally been propagated by seeding or division. Plants of the genus Asparagus are dioecious, and male plants are said to be better in terms of quality and cultivation management. Furthermore, since it is a crop with a large amount of genetic variation, propagation by sowing leads to the contamination of female plants and causes differences in yield and quality between plants, which poses a significant problem in cultivation. In addition, propagation by division requires many hands and a long period of time, is extremely inefficient, and has a high risk of transmitting viral diseases. In order to improve the above-mentioned problems, mass propagation of superior strain clones using U-woven culture has recently attracted attention. Usually, for mass propagation of asparagus, Mi strands are cultured, a large number of shoots are propagated, and then each shoot is transplanted to a rooting medium, and seedlings are formed through differentiation of adventitious roots. but,
The problem is that the rooting rate is generally low, so the propagation efficiency is low. In addition, attempts have been made to culture tissue pieces and form calluses, and then use somatic embryos produced from the calluses to achieve mass proliferation (for example, in 1981
Abstracts of research presentations at the Autumn Conference of the Horticultural Society p210-211,
November 23-25, 1986, Ryukyu University, 1988
Annual Horticultural Society Research Presentation Abstracts p254-255, 1988
．． 10.7-9, at Kyushu University). Although growing somatic embryos solves the problem of low rooting rate, the problem is that most isolated somatic embryos develop callus or malformation during culture. [Problems to be Solved by the Invention] The present inventors recognized that the conventional tissue culture method for Asparagus plants has the above-mentioned problems, and developed a tissue culture method using a novel method different from the conventional method. We investigated a method for efficiently propagating seeds and seedlings of plants of the genus Asparagus.

[Means to solve the problem]

As a result, the present inventors have found that when propagating Asparagus plants through tissue culture, the problems of the above-mentioned conventional techniques can be satisfactorily solved by propagating the plants via tuber-like tissues. Based on this new knowledge, further research has led to the completion of the present invention. Therefore, according to the method of the present invention, (1) a method for forming a tuber-like tissue of a plant of the genus Asparagus, which comprises culturing tissue or callus of a plant of the genus Asparagus to form a tuber-like tissue; The method for forming a tuber-like tissue of a plant of the genus Asparagus described in ■, the method described in ■ or ■, characterized in that the plant tissue is a tuber-like tissue obtained by culturing the tissue or callus of a plant of the genus Asparagus. The sections obtained by cutting the formed tuber-like tissue are further cultured to enlarge the tuber-like tissue,
Furthermore, if necessary, similar tuber-like tissue cutting → section culture →
A method for propagating tuber-like tissues of plants of the genus Asparagus, characterized by repeating the multiplication process of tuber-like tissue hypertrophy; A method for propagating a plant of the Asparagus genus, ■ a step of culturing tuber-like tissue and growing a plant;
The method for propagating a plant of the Asparagus genus according to (1), which is a step of culturing a tuber-like tissue to regenerate the tuber-like tissue, and culturing the tuber-like tissue to grow a plant; Provided is a method for propagating seeds and seedlings of plants of the genus Asparagus, which comprises acclimatizing the plants obtained by method (1) or (2) to produce seedlings. The invention will be explained in detail below.

In the present invention, any plant belonging to the genus Asparagus can be used. Specifically, the plant is Asparagus officinalis, which is an edible asparagus.
L. var. altjlis L. It is preferable to use this in the present invention. In the present invention, first, tissue or callus of a plant of the genus Asparagus is cultured to form a tuber-like tissue. Examples of tissues used for tissue culture in this case include tissues such as embryos, shoot tips, stems, pseudoleaves, and rhizomes. Regarding callus, in the present invention, callus obtained by culturing the tissue by a known culture method (for example, Horticultural Society Journal, Vol. 40, No. 4, pp. 11-17, 1971) can be used.

Here, the tuber-like tissue refers to a part of the underground stem (crown) that has enlarged and lengthened and accumulated storage substances.

The tissue for forming this tuber-like tissue is preferably a crown obtained by culturing the above-mentioned tissue or callus.

In this case, the crown as used in the present invention corresponds to the underground stem of the whole plant, and is a storage tissue that is morphologically equivalent to a shortened stem, and has a lump-like or crown-like shape obtained by tissue culturing Mi weave or callus. It refers to a mass of cells that contain buds or bud primordia that form the basis of buds. The medium used for tuber-like tissue formation of the present invention contains inorganic substances and carbon sources as essential components, to which plant hormones and vitamins are added, and amino acids are further added as necessary. It is. Inorganic components of the medium include inorganic salts containing elements such as nitrogen, phosphorus, potassium, sodium, calcium, magnesium, sulfur, iron, manganese, zinc, boron, molybdenum, chlorine, iodine, cobalt, etc. Specifically, potassium nitrate, sodium nitrate, ammonium nitrate, potassium chloride, calcium chloride, potassium monohydrogen phosphate, sodium dihydrogen phosphate, magnesium sulfate, magnesium chloride, sodium sulfate, ferrous sulfate, ferric sulfate, sulfuric acid. Examples include compounds such as manganese, copper sulfate, sodium molybdate, molybdenum trioxide, potassium iodide, zinc sulfate, boric acid, and cobalt chloride.

Examples of carbon sources for the medium include carbohydrates such as sucrose and glucose, derivatives thereof, organic acids such as fatty acids, and primary alcohols such as ethanol.

Examples of the plant hormones in the medium include naphthalene acetic acid (NAA), indole acetic acid (rAA), P-chlorophenoxyacetic acid, 2,4-dichlorophenoxyacetic acid (2
．． 4-D), indolebutyric acid (IBA), and auxins such as derivatives thereof, and penzyladenine (BA)
Examples include cytokinins such as , kinetin, and zeatin.

The vitamins in the medium include biotin, thiamine (vitamin 81), pyridoxin (vitamin B.), pyridoxal, pyridoxamine, calcium bantothenate,
ascorbic acid (vitamin C), inositol, nicotinic acid, nicotinamide and riboflavin (vitamin B
6) etc. can be exemplified. The main amino acids of the medium are:
For example, glycine, alanine, glutamine, cysteine,
Examples include phenylalanine and lysine. The medium of the present invention usually has a mineral content of about 0.1
μH to about 100mM, the plant hormones to about 0
．． 01 ■/l to about 150 ■/l and the above amino acids from O to about 1000+ag/j1! It is preferable to include it in use. Specifically, the medium used in the U-woven culture according to the present invention includes a conventionally known culture medium used in plant tissue culture, such as Murashige Skoog ('62
) [Murashige &Skoogl's medium,
Linsmeyer Skoog (RM-1965) [Li
ns@aier&Skoog] medium, white (
'63) [White] medium, Gamborg [Gam
borg]'s B-5 medium, Mitsui's Mon 9 medium, 27-chi.-
An example is a medium prepared by adding the above-mentioned carbon sources and plant hormones to Nitch & Nttchl, etc., and further adding the above-mentioned vitamins and amino acids as necessary. In terms of inventions, Niche Ecchi, Linsmeyer Skoog, and Murashige
A medium prepared using Skoog's medium is preferred. Regarding the composition of the above-mentioned conventionally known culture medium, see, for example, "New Plant Tissue Culture," by Takeuchi, Nakajima, and Furuya, p.386-p.3.
91 Asakura Shoten, 1979. The medium that can be used in the present invention is a liquid medium or a solid medium containing usually 0.1 to 2% of a gelling agent such as agar or Gelrite. In the present invention, tissue or callus of an Asparagus plant is tissue cultured using the above-mentioned medium to form a tuber-like tissue,
The tuber-like tissue is tissue cultured to grow a plant body, or the tuber-like tissue is further grown by tissue culture of the cut section obtained by cutting the tuber-like tissue, and if necessary, a similar method is grown. By repeating the multiplication process of cutting tuber-like tissue → section tissue culture → expanding tuber-like tissue, the grown tuber-like tissue is tissue cultured and a plant is grown. For the growth of tuber-like tissue, a medium similar to that used in the above-mentioned tuber-like tissue formation is used.

In the above-mentioned plant growth process, a crown can be regenerated from a tuber-like tissue, and the crown can be tissue cultured to regenerate a plant.

The plant growth process described above includes the rooting process. Examples of media used to grow plants include:
Murashige Skoog ('62) [Murashige
&Skoogl's medium, Linsmeyer-Skoog (
RM-1965) [Linsmaier & Sko
ogl medium, white ('63) [Wh i
te] medium, Gamborg [Gasaborg]
B-5 medium of Mitsui, M-9 medium of Mitsui, Nitch & Nitch medium, etc., the carbon source and plant hormones described above are added, and if necessary, the vitamins and amino acids described above are added. An example of this is a medium prepared by adding .

Furthermore, in the present invention, seedlings of plants of the genus Asparagus can be obtained by acclimating the plants.

Next, each method of the present invention and the tuber-like tissues, plants, etc. obtained by each method will be explained based on the drawings.

FIG. 1 is a diagram showing processes such as tuber-like tissue formation, proliferation, and plant growth of plants of the genus Asparagus according to the method of the present invention. The upper part of the figure shows the route by which a crown is formed by culturing the sections obtained by cutting the shoot stem, and the lower left part of the figure shows the path through which the tuber-like tissue is grown from the crown. The growth cycle of formation and multiplication is shown, and the lower right side of the figure shows a plant that has been rooted and regenerated from the tuber tissue grown in this way.

〔Example〕

Hereinafter, the method of the present invention will be specifically illustrated by examples.

Example 1 The crown of the rooting seedling of the asparagus variety "Hokkai 100" maintained in a sterile system was cut into pieces containing one bud each. The medium contains inorganic elements
S medium, organic elements based on etch/niche, NH4
NO3 was increased to 1/2, CaC1. was doubled and 10-'M glutamine was added to the basal medium (hereinafter referred to as AS
P medium) and shi! F sugar 60g/1, Gelrite 2.5g/l, pH 5. 7, and add lBA as auxin to 3 XIO-'M and BA as cytokinin to 10-'M, 3XIO-'M, 1
0-'M, 10-'M, uncimidol as anti-gibberellin.
'? I was added. After dispensing 3 Qae each into commercially available mayonnaise bottles (volume approximately 220 ml'), place 5 of the above asparagus crown slices and fill the mayonnaise bottles with 100ml.
When the cells were cultured for 8 weeks at 25° C. and 30,001 ux, the results shown in Table 1 were obtained.

Example 2 The tuber-like tissue of the asparagus cultivar "Hokkai 100" grown by the method of Example 1 was cut into pieces containing 2 to 3 buds. The medium is based on ASP medium,
Shi = s t46og7 x, gelrai 1.7g/i! ，
The pH was adjusted to 5.7, and 3 XIO-'M of ancymidol was added as an anti-gibberellin. After dispensing 30 pieces of each into commercially available mayonnaise bottles (capacity: approximately 220 d), 5 pieces of each of the above-mentioned asparagus tuber-like tissue sections were placed and cultured in 10 mayonnaise bottles at 25°C under 30,001 ux for 8 weeks. As a result, the results shown in Table 2 were obtained.

Example 3 The tuber-like tissue of the asparagus cultivar "Hokkai 100" maintained by the method of Example 2 was cut into pieces containing 2 to 3 buds and used as a material. The medium is basically ASP medium, and SigI! 10, 30. 60g/l, pt{5
．． 8, and after adding 10 mg of Anshidole as an anti-gibberellin, this was poured into a commercially available mayonnaise bottle (capacity: approximately 2).
20mg, one fluorocarbon membrane filter diameter Low on the lid) about 0.9g of polyester wool
After dispensing 40 Id each, five of the above asparagus tuber-like &ll tissue sections were placed on a bed and cultured in 10 mayonnaise bottles at 25°C and 30,001 ux for 8 days, and the results shown in Table 3 were obtained. Table 1 Table 3 BA concentration 10-'M 3X10-'10-'M Tuberous tissue formation rate 26% 50% 45% Sitose concentration 10g/1. 30g/ 1 60g/ 1 Rooting rate 10% 35% 44% Table 2 Multiplication rate 4.5 times [Effects of the invention] According to the method of the present invention, tuber-like tissues are formed from Asparagus plants by culturing. Furthermore, this can be efficiently propagated in large quantities, and furthermore, plants and seedlings can be efficiently propagated in large quantities via the obtained tuber-like tissues.

[Brief explanation of the drawing]

FIG. 1 is a diagram showing processes such as tuber-like tissue formation, proliferation, and plant growth of plants of the genus Asparagus according to the method of the present invention. FIG. 2 is a photograph showing the tuber-like tissue of asparagus formed by the method of the present invention. Furthermore, Figure 2 is a photograph showing the morphology of living things. Figure 1

Claims (1)

[Scope of Claims] 1. A method for forming a tuber-like tissue of a plant of the genus Asparagus, which comprises culturing tissue or callus of a plant of the genus Asparagus to form a tuber-like tissue. 2. The method for forming a tuber-like tissue of a plant of the genus Asparagus according to claim 1, wherein the tissue of the plant of the genus Asparagus is a crown obtained by culturing tissue or callus of a plant of the genus Asparagus. 3. The sections obtained by cutting the tuber-like tissue formed by the method according to claim 1 or claim 2 are further cultured to enlarge the tuber-like tissue, and if necessary, the same tuber-like tissue is cut → A method for growing tuber-like tissue of plants of the genus Asparagus, which is characterized by repeating the growth process of section culture → tuber-like tissue hypertrophy. 4. A method for propagating a plant of the genus Asparagus, which comprises growing a plant by culturing the tuberous tissue obtained by the method according to any one of claims 1 to 3. 5. The step of culturing the tuber-like tissue and growing the plant body,
5. The method for propagating a plant of the genus Asparagus according to claim 4, comprising the steps of culturing a tuber-like tissue to regenerate a crown, and culturing the crown to grow a plant. 6. The method for propagating a plant of the genus Asparagus according to claim 4 or 5, wherein the growing step of the plant includes a rooting step. 7. A method for propagating seeds and seedlings of plants of the genus Asparagus, which comprises acclimating the plants obtained by the method according to claims 4 to 6 to produce seeds.