JP3591250B2 - Tide-resistant plant and method for producing the same - Google Patents

Description

[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a plant having tide resistance and a method for producing the same. Since plants having tide resistance have high resistance to sea breeze, they enable greening of beach areas, landfills, and the like, which were difficult to green.
[0002]
[Prior art]
Beach areas are often exposed to sea breeze, and in the case of strong winds, a large amount of seawater blown by the wind adheres to plants, and plants with low tide resistance often die. When greening a beach area, a tide net is set up, and tidal resistant native species are selected and planted.
[0003]
On the other hand, methods for improving the salt tolerance of plants have also been developed. JP-A-8-193018 discloses a technique for alleviating salt stress in plants by applying a supernatant (brown juice) discharged when extracting green leaf protein from alfalfa to soil where the plants are grown. ing.
[0004]
[Problems to be solved by the invention]
The method of installing a tide net is not a fundamental solution because the appearance is bad and the wind containing seawater may leak in strong winds. Furthermore, in the method of planting a native plant having strong tide resistance, a plant that can be used for planting is limited, so that a free planting plan cannot be made.
Further, the method described in Japanese Patent Application Laid-Open No. 8-193018 requires a facility for applying brown juice on a regular basis, and there is a concern about an increase in maintenance costs.
[0005]
The present invention has been made under such a technical background, and an object of the present invention is to provide a tide-resistant plant and a method for efficiently producing a tide-resistant plant.
[0006]
[Means for Solving the Problems]
The present inventors have conducted intensive studies to solve the above-mentioned problems, and as a result, found that plants having high tide resistance occur at a high frequency by performing polyploid expression treatment of plants, and completed the present invention from this finding. did.
[0007]
That is, the present invention is a plant having tide resistance, which contains a tissue having a chromosome doubled as compared with a conventional variety in a plant body.
Further, the method for producing a plant, wherein the plant is subjected to polyploid expression treatment, a plant having a doubled chromosome is selected from the treated plants, and then a plant having tide resistance is selected from the selected plants. It is.
[0008]
BEST MODE FOR CARRYING OUT THE INVENTION
(1) The plant of the present invention has the following characteristics.
It is similar in appearance to the plant before improvement, but can be distinguished from the fact that it contains tissues with doubled chromosome numbers. This can be determined from the size of guard cells without calculating the number of chromosomes. It can also be distinguished from the fact that leaf damage (salt stress) caused by salt adhering to the aerial part is reduced as compared to the plant before improvement.
In addition, the above-mentioned tissue whose chromosome has been doubled includes not only a plant in which all the tissues of the plant have doubled, but also a plant in which only a part of the tissues has doubled.
[0009]
(2) Method for producing the plant of the present invention The plant of the present invention is obtained by subjecting a plant to a polyploid expression treatment, selecting a plant containing cells with a doubled chromosome number from the treated plants, and then selecting a plant having tide resistance Can be created by selecting
[0010]
Examples of plants that carry out polyploid expression processing include the genus Echinida, the genus Cotoneaster, the genus Mice, the genus Acacia, the genus Quercus, the genus Zeus, the camphor, the gardenia, the azalea, the camellia, the tovera, and the eucalyptus. Can be.
[0011]
Among them, as the genus Enishida, plants such as Enishida, Houbenienisida, and Cutienienida are preferable, and as the Cotoneaster genus, plants such as Salix toxin, Benishitan, and Himesharinto are preferable. Plants such as hornworm and rodent are preferable, and as the genus Acacia, plants such as Husa acacia and Ginkgo acacia are preferable.As the genus Quercus, plants such as ubamegashi, arakashi and shirakashi are preferable. Plants are preferable, as the Camphor tree, camphor trees, plants such as Yabnikkei are preferable, as the gardenia, plants such as gardenia, gardenia, etc. are preferable, and as the azalea genus, plants such as Hirado azalea, Kirishima azalea are preferable, and as the Camellia genus Is the Japanese camellia, Saza Preferably plants such as mosquitoes, the Pittosporum, preferably plants such as Pittosporum tobira, as the eucalyptus, eucalyptus, although plants are preferred, such as delicious NARIS eucalyptus, is not limited to a particular plant.
[0012]
The polyploid expression treatment includes a method using a polyploid inducing agent, a method using radiation such as gamma rays, X-rays, and ultraviolet rays, and a method using these in combination. From the aspect of sex, it is desirable to use only a polyploid inducer.
[0013]
Examples of polyploid inducers include colchicine, laughing gas, colcemide, acenaphthene, vinblastine, bodophilotoxin, coumarin, atropine, veratrine, nicotine, sanguinarine, benzol derivative, diphenyl derivative, phenanthrene derivative, naphthalene derivative, diphenylamine, triphenylamine. Bromaniline, paradichlorobenzol, methylnaphthoquinone, methylnaphthohydroquinone, salicylic acid and similar substances, hexachlorohexane, pyruvine and its hydrochloride, alkyl-alkali metal carbamates, phenylurethane, cacodylate, convalarin, convalatoxin, con Examples include, but are not limited to, balamarin, heteroauxin, germisan, chloroform, amiproposmethyl, oryzalin, and the like. , May be any as long as the agent has the effect of doubling the chromosomes.
[0014]
As a specific treatment method, for example, a method of immersing cultured cells such as seeds, rhizomes, or callus in a medium containing a polyploid inducer for a certain period of time can be exemplified, but any method capable of doubling chromosomes can be used. However, it is not limited to these. The most preferable treatment method is a method in which seeds are immersed in a medium containing a polyploid inducer for a certain period of time.
[0015]
When colchicine is used as the polyploid-inducing agent, the concentration of colchicine is preferably 0.001-2%, more preferably 0.05-0.1%. In addition, the immersion time is preferably set to 1 to 10 days, more preferably 4 to 5 days.
[0016]
After the polyploid expression treatment, plants containing cells whose chromosome number has been doubled are selected from the plants. Examples of the selection method include a method of collecting cells from a plant and measuring the amount of DNA, and a method of measuring the number of chromosomes at the root tip.
In general, it is known that a doubled plant has a larger cell volume than a plant before the polyploid expression treatment, so that leaves and petals become thicker and the guard cells increase in size. The presence or absence of doubling may be determined by measuring the size of doubling to select a doubled plant.
[0017]
Further, from the plants selected as described above, plants having tide resistance are selected. The presence or absence of tide resistance is determined by spraying a fixed concentration of sodium chloride solution onto the aerial part of the plant and then examining the degree of leaf damage (salt stress).
In the case where the degree of the damage (salt stress) is determined by quantifying the ratio of the leaves that have changed color due to the damage to the entire leaf of the plant, the degree of the damage is smaller than that of the plant before the treatment. Less than half of the plants are judged to be tidal resistant plants.
By the method as described above, the tide-resistant plant of the present invention can be produced.
[0018]
(3) Method of Using the Plant of the Present Invention The plant of the present invention has tide resistance, so that it can be used as a greening tree in a beach area.
[0019]
Embodiment 1
Hereinafter, the present invention will be described in more detail with reference to examples.
The seeds of Cysticus scoparius were absorbed for 5 days on a filter paper impregnated with a 0%, 0.05% or 0.1% aqueous solution of colchicine, washed, and sown on vermiculite culture soil. About 100 seeds were used for each condition. An individual in which the size of guard cells was significantly increased among the individuals that germinated three months later was determined to be an individual containing cells whose chromosome number was doubled.
With 0.05% colchicine treatment, 6/28 germinated individuals were obtained, and with 0.1% colchicine treatment, 2/29 individual cells containing doubled cells were obtained.
[0020]
Individuals not treated with colchicine (diploid: 1a, 1b, 1c) and individuals containing cells with doubled chromosome numbers (line: 1A, 1B) are propagated by cuttings, and dividing cells at the root tip of each line Was collected, and the number of chromosomes was calculated. In addition, the observation of the chromosome was performed by the orcein acetate-crush method.
[0021]
The number of chromosomes in the line not treated with colchicine was 2n = 48 in the diploid form. On the other hand, among the lines containing cells whose chromosome number was doubled, line 1A had 2n = 48 and line 1B had 2n = 96. From these results, line 1A determined that the L1 layer in which the epidermis differentiated was a tetraploid, the L3 layer in which the lateral roots differentiated was a diploid peripheral chimera, and line 1B was an autotetraploid.
[0022]
The tide resistance was measured using three diploid lines and two lines containing doubled cells. The measurement of tide resistance was determined by spraying a 600 mM aqueous sodium chloride solution over the entire above-ground part and counting the number of injured leaves one month later. In each line, the percentage of damaged leaves was 27.2% for 1a, 36.8% for 1b, 33.8% for 1c, 18.8% for 1A, and 11.8% for 1B. (FIG. 1).
[0023]
Both line 1A and line 1B were less injured by sodium chloride than the diploid line, and particularly in line 1B, the damage was about 1/3 of the average value of the diploid line.
From the above results, it was found that by producing a plant containing the doubled cells, a tide-resistant enishida can be efficiently produced.
[0024]
Embodiment 2
Cotoneaster salicifolius seeds were allowed to absorb water for 5 days on a filter paper impregnated with a 0% or 0.1% aqueous solution of colchicine, washed and then sown on vermiculite culture soil. About 100 seeds were used for each condition. Three months later, an individual in which the size of guard cells was significantly increased among individuals that germinated was determined to be an individual containing cells whose chromosome number was doubled. Two individuals out of ten individuals germinated by the 0.1% colchicine treatment were obtained containing individuals that had doubled cells.
[0025]
Individuals not treated with colchicine (diploids: 2a, 2b, 2c) and individuals containing cells with doubled chromosomes (lineages: 2A, 2B) are propagated by cuttings, and dividing cells at the root tip of each lineage Was collected, and the number of chromosomes was calculated in the same manner as in Example 1.
[0026]
The number of chromosomes of the line not subjected to the colchicine treatment was 2p = 34 (diploid type). On the other hand, among the lines containing cells whose chromosome number was doubled, line 2A was 2n = 34 and line 2B was 2n = 68.
[0027]
From these results, line 2A determined that the L1 layer in which the epidermis differentiated was a tetraploid, the L3 layer in which the lateral roots differentiated was a diploid peripheral chimera, and that line 2B was an autotetraploid.
Tide resistance was measured using three diploid lines and two homotetraploid lines. The measurement of tide resistance was determined by spraying a 600 mM aqueous sodium chloride solution over the entire above-ground part and counting the number of injured leaves one month later. In each line, the percentage of damaged leaves was 36.1% for 2a, 47.8% for 2b, 48.1% for 2c, 21.1% for 2A, and 24.9% for 2B. (FIG. 2).
[0028]
In both the lines 2A and 2B, the damage caused by sodium chloride was smaller than that in the diploid line. In the line 2A, the damage was less than half of the average value of the diploid line.
From the above results, it is possible to efficiently produce tide-resistant cotoneaster by producing a plant containing the doubled cells.
[0029]
Embodiment 3
The seed of Ligustrum lucidum was absorbed on a filter paper impregnated with an aqueous solution of 0% or 0.1% colchicine for 4 or 5 days, washed, and then sown on a vermiculite culture soil. About 100 seeds were used for each condition. Three months later, an individual in which the size of guard cells was significantly increased among individuals that germinated was determined to be an individual containing cells whose chromosome number was doubled.
[0030]
With 0.1% for 4 days of colchicine treatment, 5 out of 22 germinated individuals and 5 days of colchicine treatment obtained 4 x 12 individuals containing cells that had doubled. Individuals not treated with colchicine (diploids: 3a, 3b, 3c) and individuals containing cells whose chromosome number has doubled (strains: 3A, 3B, 3C, 3D) are grown by cuttings, and the roots of each strain are grown. The dividing cells at the end were collected, and the number of chromosomes was calculated in the same manner as in Example 1.
[0031]
The number of chromosomes in the line not treated with colchicine was 2p = 46 in the diploid form. On the other hand, among the lines containing cells whose chromosome number was doubled, lines 3A and 3B had 2n = 46, and lines 3C and 3D had 2n = 92. From these results, in line 3A and line 3B, the L1 layer in which the epidermis differentiates is a tetraploid, and the L3 layer in which the lateral root differentiates is a diploid peripheral chimera, and lines 3C and 3D are allotetraploid. Was determined.
[0032]
Tide resistance was measured using three diploid lines and four lines containing doubled cells. The measurement of tide resistance was determined by spraying a 600 mM aqueous sodium chloride solution over the entire above-ground part and counting the number of injured leaves one month later. In each line, the percentage of affected leaves was 29.6% for 3a, 31.0% for 3b, 56.0% for 3c, 41.7% for 3A, 11.4% for 3B, and 3C for 3B. Was 8.3% and 3D was 50.6% (FIG. 3).
[0033]
The degree of damage of the line containing the doubled cells varied greatly between the lines, and both lines 3B and 3C were less injured by sodium chloride than the diploid line, and １ ／ of the average value of the diploid line. It was a degree of injury.
By producing a plant containing the doubled cells by the above-mentioned method, it is possible to efficiently produce a tide-resistant porphyra hornworm.
[0034]
【The invention's effect】
The present invention provides a novel plant and a method for producing the same. Since the plant of the present invention has tide resistance, it can be used for planting in a beach area. Further, the method for producing a tide-resistant plant of the present invention can be efficiently produced with less labor required for production.
[Brief description of the drawings]
BRIEF DESCRIPTION OF DRAWINGS FIG. 1 is a graph showing the tide resistance of a diploid strain and a tetraploid strain.
FIG. 2 is a graph showing the tide resistance of a diploid cotoneaster and a tetraploid cotoneaster.
FIG. 3 is a graph showing the respective tide resistances of a diploid strain of the mouse and a tetraploid strain of the mouse.

Claims (2)

Treating the plant with polyploid expression,
From the plants after the treatment, select a plant containing a tissue whose chromosome has doubled,
From the plant containing the selected chromosome doubled tissue, to select a plant having tide resistance,
A method for producing a tide-resistant plant, comprising: The method for producing a tide-resistant plant according to claim 1, wherein a plant having an increased guard cell size is selected as a plant containing the tissue whose chromosome has been doubled.

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