JPH04335839A - Method for promoting morphogenesis or differentiation of plant - Google Patents

Abstract

PURPOSE:To provide a method for improving the morphogenesis or differentiation frequency in the formation or differentiation of new organ or embryoid body from tissue or cell of a plant. CONSTITUTION:The morphogenesis or differentiation of a plant is promoted by adding an alginic acid oligosaccharide or its heat-treated product into a medium in the culture of the tissue or cell of a plant.

Description

[0001] The present invention relates to a method for promoting morphogenesis or differentiation of plants, and more specifically, the present invention relates to a method for promoting morphogenesis or differentiation of plants, and more specifically, the present invention relates to a method for promoting morphogenesis or differentiation of plants, and more specifically, for producing new plant organs or embryoid bodies from plant tissues or cells. When selecting and maintaining plants that possess specific traits that cause them to form, and when producing them as seedlings, it is possible to enhance the formation of plant organs or embryoid bodies by adding specific components to the culture medium of the plant's tissues or cells. The purpose is to promote the efficient production of seeds and seedlings useful in agricultural production. BACKGROUND OF THE INVENTION [0002] Improvement of plant seeds and seedlings is an important issue in agriculture, and has been energetically tackled since ancient times. In recent years, advances in genetic engineering, cell fusion technology, tissue culture technology,
Advanced technologies such as advances in robotics have entered the stage of practical application, and the production of useful plant seeds and seedlings for agriculture and horticulture has begun using so-called bionursery systems that combine such advanced technologies. The most important step in such seedling production methods is to culture target plant tissues and cells to form or differentiate new organs and the like. Traditionally,
In addition to medium composition, temperature, pH, etc., various conditions such as application concentration and addition timing of plant hormones such as auxin and kinetin have been investigated. However, as a result of these studies, even under the set conditions, the rate of organ formation and the frequency of differentiation are not sufficient for practical purposes, and the development of means to promote the rate of formation and differentiation of plant organs, etc. was long awaited. [Means for Solving the Problems] An object of the present invention is to discover a substance that promotes the formation rate and differentiation of plant organs, and to apply this substance to the production of plant seeds and seedlings to improve production efficiency. It is to aim for it. As a result of extensive research to search for such substances, the inventors discovered that alginic acid oligosaccharides are effective, and have arrived at the present invention. That is, the present invention provides a method for promoting plant morphogenesis or differentiation, which comprises adding alginate oligosaccharide or a heat-treated product thereof to a medium when culturing plant tissues or cells. [0005] The alginate oligosaccharide used in the present invention can be obtained by decomposing alginic acid, sodium alginate, alginate-containing algae such as kelp, or microbial polysaccharide with an enzyme such as alginate lyase, or hydrolyzing with an acid such as hydrochloric acid. This is an oligosaccharide composition obtained by The main components of the oligosaccharide are guluronic acid, mannuronic acid, and 4,5-deoxyuronic acid. An oligosaccharide composition is a composition having a degree of polymerization of 2 to 20 and consisting of an oligosaccharide containing guluronic acid and/or mannuronic acid, or these and 4,5-deoxyuronic acid, or a composition in which these are heated. Processed products, specifically oligosaccharides composed only of guluronic acid, oligosaccharides composed only of mannuronic acid, oligosaccharides composed of guluronic acid and 4,5-deoxyuronic acid, and guluronic acid. An oligosaccharide composed of mannuronic acid and mannuronic acid, mannuronic acid and 4,5-
Oligosaccharide composed of deoxyuronic acid, guluronic acid,
Oligosaccharides composed of mannuronic acid and 4,5-deoxyuronic acid, or those obtained by heat-treating these at pH 1-8 and temperature 90-130°C for 15-180 minutes. Can be done. Next, the plants to which the present invention is applied are not particularly limited, and include vegetables such as potatoes, tomatoes, cabbage, carrots, and lettuce, grains such as rice, wheat, and corn, and flowers such as lilies. It can be widely applied to fruit trees such as mandarin oranges and apples. Oligosaccharide alginate or its heat-treated product is usually added to the medium used for culturing plant tissues or cells.
．． It is used by adding it at a concentration of 0.01 to 5000 ppm, but in another embodiment, the plant tissue or cells are immersed in a solution containing this substance, or the solution is directly applied to the plant tissue or cells. It is also possible to use [0008] Except for using alginic acid oligosaccharide or a heat-treated product thereof, plant tissues or cells may be cultured under known conditions to achieve morphogenesis or differentiation of the desired plant. [Example] Next, the present invention will be explained with reference to Examples, but the present invention is not limited thereto. Example 1 Production of alginate oligosaccharide After dissolving 10 parts of water per 1 part of sodium alginate,
The pH was adjusted to 7.0, and 3000 units of alginate lyase was added per 1 g of sodium alginate to perform an enzyme reaction. In addition, the enzymatic activity of alginate lyase is at pH 7.0.
, When the enzyme was applied to a 0.2% sodium alginate solution at 30°C, the absorbance at 230 nm decreased to 0.01 in 30 minutes.
The enzyme power to be increased was defined as 1 unit. The enzyme reaction was carried out at 30°C for 24 hours, and after the reaction, the pH was further adjusted to 3.0.
Heat treatment was performed at 00°C for 60 minutes. Through the above operations, alginic acid oligosaccharides containing 70% or more of oligosaccharides having a degree of polymerization of 2 to 20 based on the solid content could be obtained. Example 2 Using potato shoots grown by potato microtuber shoot apical culture,
The effect of alginate oligosaccharides on the number of microtubers formed was investigated. [0011] Shoots are mixed with 3% sucrose and 0.8%
After growing in Murashige-Skoog medium (hereinafter referred to as MS medium) containing agar, 5% sucrose, 5 ppm benzyladenine, 0.8% agar and various concentrations of alginate oligosaccharide prepared in Example 1 were grown. Microtubers were induced with MS medium containing Culture 1
Table 1 shows the number of microtubers with a diameter of 3 mm or more formed and the fresh weight after a month. Note that () in the table is an index relative to the control. 0012
Table 1 Concentration (ppm) Number of tubers formed (pieces/lot) Total fresh weight (g)
0 10 (100)
3.1 (100
) 6
11 (110)
3.3 (106) 60
19 (190)
5.2 (168)
600 18 (180)
4.3 (139)
Example 3 Cultivation of lily scales Bulblets derived from lily scales were cultured.
The effect of alginate oligosaccharide on shoot differentiation was investigated. [0014] Bulblet was mixed with 3% sucrose and 0
．． The cells were induced in MS medium containing 8% agar and cut into 5 mm square sections. This section was cultured in MS medium containing 3% sucrose, 0.1% kinetin, 0.1 ppm NAA, 0.8% agar, and various concentrations of the alginate oligosaccharide prepared in Example 1. Table 2 shows the number of stem and leaf differentiation after two months of culture. Note that ( ) in the table is an index relative to the control. [0015]
Table 2 Concentration (ppm) Number of stem/leaf differentiation (plants/lot)
0
2 (100)
0.025
12 (600)
2.5
24 (1200)
250 29
(1450) Example 4 Stem and leaf differentiation of tobacco callus The influence of alginate oligosaccharide on the differentiation of tobacco callus into leaf and stalk was investigated. Tobacco (Nicotiana)
tabacum) in a growth medium (1.0 ppm).
MS containing NAA, 0.1 ppm BA and 0.8% agar
The callus was cultured in a medium (medium), and the calli on the 20th day of subculture were used for differentiation into stems and leaves. [0017] The callus was cut into approximately 1 cm square sections, and a differentiation medium (0.1 ppmNAA, 1.0 ppm BA, 0.8%
The cells were cultured for about one month on MS medium containing agar and alginate oligosaccharide prepared in Example 1 at various concentrations, and the number of stem and leaf differentiation was compared. The results are shown in Table 3, and the values are expressed as the number of stem and leaf differentiation per callus using 10 pieces of callus from each section. Note that ( ) in the table is an index relative to the control. [0018]
Table 3 Concentration (ppm
) Number of stem and leaf differentiation (pcs/lot)
0
1.6 (100)
0.1
3.1 (194)
1.0 3.9
(244)
10 4.7 (29
4) 100
4.5 (281)
1000
4.4 (275)
5000
4.1 (256) [Effects of the Invention] When culturing plant tissues or cells to form or differentiate new organs, etc., by adding alginate oligosaccharide or a heat-treated product thereof to the medium. , it is possible to promote the morphogenesis and differentiation of plants and improve the production efficiency of artificial seeds and seedlings of useful agricultural and horticultural crops.

Claims (2)

[Claims] 1. A method for promoting plant morphogenesis or differentiation, which comprises adding alginate oligosaccharide or a heat-treated product thereof to a medium when culturing plant tissues or cells. Claim 2: The alginic acid oligosaccharide has a polymerization degree of 2 to 2.
2. The method according to claim 1, wherein the oligosaccharide is an oligosaccharide whose constituent components are guluronic acid and/or mannuronic acid, or these and 4,5-deoxyuronic acid.