JP3484166B2 - Early detection of woody plant seeds - Google Patents

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an early test method for woody plant seeds, an early quality proof method for plant seeds using the early test method, and a germination rate obtained by these methods. Design method for setting the seeding rate of woody plant seeds in Japan. This method is used for normal germination tests such as seeds with slow embryo growth such as woody plants, seeds with a slow germination rate because they have physiological dormancy, and seeds with a slow germination rate because they are solid seeds. Apply to difficult seeds. In particular, a method for hardwood seeds is provided.

[0002]

2. Description of the Related Art Seeds have various structures depending on the species, but in the present specification, seeds are basically described as organs composed of the following three tissues. 1) Embryo: Radicle, Hypocot
y1), Cotyledon (Cotyledon), Plumule (apical meristem), Epicotyl (Epicoty1)
Consists of (Stem). 2) Endosperm: Depending on the species, endosperm (Endospem)
rm) rather than ectosperm (Perisperm). 3) Seed-coat: Each seed is composed of at least two layers, but in the present specification, inner seed-coat (including endosperm developed) (Inner seed-coat or Endopleura) and outer seed-coat (( Depending on the seed, pericarp or pseudoseed (Ar
il))) (Outer seed-coat or Testa). Depending on the species, the seed coat is more mucous (polysaccharide)
It may also be wrapped in lipids (wax).

[0003] In the case of a tree planting (seeding) technique for treating seeds of woody plants, in measuring the germination rate of seeds of woody plants, the seeds are usually arranged on a petri dish lined with one or two moistened filter papers, and the seeds are regularly prepared. The method of counting the number of germinated individuals to obtain the germination rate is generally used. In this case, the test temperature is a constant temperature of 23 to 25 ° C. or a temperature change treatment depending on the species, and the test period is 14
~ 28 days. In addition, for seeds that do not germinate even after the test period, the cause of non-germination such as whether the seeds are cut open to be solid seeds or non-solid seeds may be investigated.

As a seed germination test method used in slope greening work and the like, the method described in "Road Earth Works-Slope Stabilization Guideline, 1999" issued by the Japan Road Association is widely used. ing. This germination test method is performed by laying two pieces of filter paper on a petri dish, arranging seeds on it, and immersing them in water, at a constant temperature of 20 to 25 ° C and at a varying temperature of 18 to 28 ° C. In the survey, the seeds that had normally sprouted are counted and removed every day, and the germination rate is calculated by cutting off the seeds in 14 days for exotic herbs and 28 days for other plants.

In addition to this germination test method, as a method for measuring the germination rate and germination power of seeds, "International Rules for Seed Testing 1985 (19) issued by the International Seed Testing Association (ISTA).
86, 1989). It describes the following germination test and germination test by embryo removal as a method for measuring the germination rate or germination ability of seeds, which is translated from the Ministry of Agriculture, Forestry and Fisheries Seedling Management Center as "International Seed Inspection Regulations". Has been issued. 1) Germination test Seeds are placed on a germination bed using paper or sand, left for 5 to 70 days under a temperature condition suitable for germination of the seeds, and then the germination rate is measured by the percentage of normal germination. 2) Germination test by embryo extraction (extracted embryo method) In order to obtain the germination force, after soaking the seeds in water for a certain period of time,
The inner seed coat is peeled off, the embryo is completely removed, and light is irradiated for at least 8 hours per day under constant temperature conditions of 20 to 25 ° C., and cultured for 5 to 14 days. As for the evaluation method, the percentage of population of embryos with germinability showing signs of growth, showing chlorophyll development, remaining firm and fresh, or coniferous embryos showing cotyledon curvature is expressed in percentage. It is expressed and evaluated by the percentage of individuals who have a viable embryo.

[0006] As a method for breaking down physiological dormancy, in the above-mentioned "International Seed Inspection Regulations", dry storage treatment (seed having a short dormancy) and pre-cooling treatment (for tree seeds, at a temperature of 1-5 ° C for 2 weeks- 12 months storage), preheat treatment (30-35 ℃)
Not over 7 days and overheated under open aeration for 7 days, light irradiation (light of 750 to 1,250 lux to test area), addition of potassium nitrate (0.2% potassium nitrate solution instead of water germination bed) ), Addition of gibberellin (GA) (adding 0.02-0.1% GA solution to the germination bed instead of water), encapsulation with polyethylene bag (at the end of the germination test, a high proportion of fresh non-germinated seeds are It is disclosed that, if found, the germination test is carried out by performing dormancy breakage by one of the methods of re-testing after encapsulation treatment, and the percentage of the number of germinated individuals is evaluated as the germination rate.

As a method for coping with the germination delay due to the hard seed, the "International Seed Inspection Regulation" states that the water content of 24 to
Soaking for 48 hours, in Acacia, soaking in boiling water three times the volume of seeds until it cools down, mechanical peeling, treatment with concentrated sulfuric acid, treatment with 1N nitric acid for 24 hours, etc. A method of performing a germination test after performing is disclosed. In addition, as another method, “Tree growth and environment, Kenichi Hatano and Yoshihiko Sasaki, 1993”, it is disclosed that germination is easily led by increasing the oxygen partial pressure.

Regarding the treatment method before embryo removal in the isolated embryo method, 5% of seeds or fruits are described in "International Seed Inspection Regulations" and "General Review of Agriculture and Forest Seed Science, Shunichiro Nakamura, 1985".
It is disclosed that it should be rinsed well with water after crushing in a sodium hypochlorite solution for 15 minutes.

[0009]

As described above, although there are many known and well-known facts about individual treatment methods and the like as a seed germination test method, the germination rate of woody plant seeds having high dormancy can be efficiently determined. A general-purpose method required in a short period of time has not been presented yet. In actual practice, it is extremely difficult to easily combine these known facts to derive an appropriate germination test method for woody plant seeds.

The germination test according to the above-mentioned "Road earthwork-Slope stabilization / slope stabilization guideline, 1999" and "International seed inspection regulations" has been frequently used since the germination rate was high, such as grasses (foreign grass). Since the herbaceous seeds for vegetation that have been developed have excellent storage stability of seeds, it is not necessary to check the germination rate of seeds before construction, and even if a germination test is performed, results can be obtained in a short period of about one week. There was no problem because he was able to guide. However, many of the seeds of woody plants, which have been increasingly used in greening work in recent years, have various dormancy properties, so the germination rate is very slow, and some of the native species do not germinate. Slower or until germination 1
There are quite a few that take ~ 2 years. Therefore, in the conventional germination test method, a large amount of mold is generated in a germination test container such as a petri dish during the measurement period, and even the healthy seeds having germination ability are rotted due to the mold generated, It was difficult to measure the exact germination rate.

In the germination test described in "International Seed Inspection Regulations", a germination test period of 1 to 2 months is required for seeds having a slow germination rate. In addition, for individuals with strong dormancy such as woody plant seeds, treatments such as long-term low temperature wet layer treatment (pre-cooling treatment) or combination of low temperature wet layer treatment and high temperature wet layer treatment are required for dormancy awakening treatment. And In addition, woody plant seeds have a low germination rate and low storability, resulting in rapid deterioration of seeds.
It was difficult to accurately measure the germination rate because the deterioration of seeds progressed after the dormant wakefulness treatment required for a long time before the germination rate was measured and before the germination test was completed.

On the other hand, in the germination test by embryo removal described in the "International Seed Inspection Regulations", it is defined as a method for judging the germination ability of a slow germination seed or a dormant seed which is difficult for a normal germination test. There is. However, this extracted embryo method requires time and effort for seed treatment, and most woody plant seeds require a measurement period of one week or longer. This method cannot be applied from the practical aspect of construction work. In particular, when it is necessary to quickly check the quality of seeds to be used and determine the seeding amount before construction due to restrictions such as the construction period, it was not applicable due to the period.

In the isolated embryo method, the seed coat is removed and the embryo is placed in a completely exposed state on seeds having any physiological and structural characteristics. However, it may be difficult to remove the seed coat or structurally difficult to remove the seed coat, as in the case of extremely small seeds. Furthermore, such a treatment is a very detailed work in practice, and it takes a considerable amount of time and labor to perform the test by treating a seed with a number of specimens corresponding to the germination test.

In addition, in the isolated embryo method, when removing outer seed coat and inner seed coat as a pretreatment for inspection, seeds having outer seed coat are immersed in water overnight to 48 hours, and then the outer seed coat is removed. It is necessary to further expose the embryo by immersing it in water overnight to 72 hours to remove the inner seed coat. Further, for seeds having only inner seed coat, it is necessary to remove the inner seed coat to expose the embryo after soaking in water for 24 to 96 hours.
Further, since a culture period after exposing the embryo is further required for 5 to 14 days, a long period of 6 to 18 days is required as a test period. It is not noted that these methods promote seed deterioration.

[0015] In the "International Seed Inspection Regulations", special instructions are written in the germination test by embryo removal, and as the handling of woody plant seeds, maple genus (excluding ash maple, Japanese maple), Rowan, euonymus, ash, apple, pear, monarch pine,
Balkan Goyou, Strobe pine, European pine, Shishima pine, Bosnian white pine, Jeffrey pine,
Embryo extraction methods are shown for Korean pine, pine pine, genus Prunus, and linden. However, these methods alone cannot cope with many kinds of woody plants currently used in greening work and the like.

[0016] As described above, the germination test method indicated by the "International Seed Inspection Regulations" for some woody seeds is not only inappropriate, but is also favored in the present greening work and the like. It does not correspond to many woody seeds, and it is a practical method when it is necessary to test the germination rate of seeds actually used before construction as early as the seeding work mainly using woody plants. I can't say.

In the recent greening work, the conventional greening mainly using greening plants represented by exotic herbs has been changed to greening using native trees growing around the construction site. Naturally collected seeds used for such construction (in some cases, such as planned collection or collection by local residents) have a germination rate that depends on the collection site, collection year, collection time, storage method after collection, etc. It fluctuates greatly. Therefore, from the practical aspect of design and construction, it is necessary to measure the germination rate of the seeds actually used before the construction to design or correct the seeding amount. In carrying out such greening work, the existing germination test methods are insufficient in terms of time and accuracy, and it is urgently necessary to establish a germination rate verification system that can be used to quickly test germination ability and utilize it for design. Therefore, it is socially required to establish it early.

Woody plant seeds not only have various structures different from herbaceous plant seeds, but also possess various dormancy patterns. Further, since the deterioration rate of seeds in the storage process is high and it is difficult to determine the vitality of the seeds in a short period of time, civil engineering construction (vegetation construction) faces great construction difficulties.

In view of the above situation, the present invention is directed to a woody plant seed, a seed pretreatment method, a seed placement method, and a seed treatment method for testing the germination rate in a short period of about 7 days or less. It is an object to provide a germination rate test method including the method. In particular, it is an object of the present invention to provide a versatile method applicable to any of various woody plant seeds currently used in revegetation and the like. Furthermore, by realizing such a germination rate test method, it is an object to meet the demands of the times when environmental problems are widely taken up, solve the above-mentioned problems, and provide a basis for rational greening technology.

[0020]

The present invention provides constant test conditions (e.g., room temperature (easy to work and economical temperature control) regardless of the structural and diapause characteristics of tree seeds (particularly hardwood seeds). It will be possible to test the average germination power or germination rate of these seed populations in a short period of about one week in the light) (around 23 ° C.). The assay method according to the present invention has versatility applicable as a systematic method according to the seed structure and dormancy of all broad-leaved tree species.
It was possible to test the germination rate and germination power around a week. The specific configuration of the present invention is as follows.

(1) In the first aspect of the early assay method for woody plant seeds of the present invention, in the early assay method for woody plant seeds, the seeds are selected according to the structural characteristics of the woody plant seeds .
At least fully immature embryo type seed, immature embryo type seed, embryo mature type
Seed, classified into any of the solid seeds, the pretreatment method of the seed, the bed placement method and the treatment method are appropriately selected according to the classification of the seed and the physiological characteristics of the seed, and the selected pretreatment. The seeds are processed by the method and the bed placement method, and the processed seeds are placed on the germination test bed under continuous light irradiation conditions and constant temperature conditions by the selected treatment method, and the seeds that are placed are germinated. The proportion of individuals with capacity and / or the germination ability is measured.

(2) In the second aspect of the early assay method for woody plant seeds of the present invention, in the early assay method for woody plant seeds, the seeds are selected according to the structural characteristics of the woody plant seeds .
At least fully immature embryo type seed, immature embryo type seed, embryo mature type
Seed, classified into any of the solid seeds, the pretreatment method of the seed, the bed placement method and the treatment method are appropriately selected according to the classification of the seed and the physiological characteristics of the seed, and the selected pretreatment. The seeds are processed by the method and the bed placement method, and the processed seeds are placed on the germination test bed under continuous light irradiation conditions and constant temperature conditions by the selected treatment method, and the seeds that are placed are germinated. measuring the rate and / or germination ability possessed individuals in advance derive a correlation between the actual germination rate of the the rate or germination of the measured germination held individual seeds, the correlation By applying, the actual germination rate of the seed is estimated from the ratio of the individuals having the germination ability or the germination ability.

[0023]

(3) In the aspect of ( 1) or (2) above, the method of placing seeds classified as the immature seeds of the complete embryo is at least as described in (1-A) to (1-F) below. It is selected from either method. (1-A): A method in which the endosperm tissue is cut so as to contain the embryo and the seed slice is placed after removing the inner seed coat. (1-B): A method in which a seed slice obtained by cutting the endosperm tissue so as to contain the embryo is placed with only the inner seed coat attached. (1-C): A method in which a seed slice obtained by cutting the endosperm tissue so as to contain the embryo is placed with the outer seed coat attached. (1-D): After cutting the endosperm tissue by any of the methods (1-A) to (1-C) above, a seed section obtained by cutting the side surface of the endosperm tissue so as to further contain an embryo is placed. Method. (1-E): A method of removing the embryo from the seed and placing it on the bed. (1-F): When it is difficult to remove the outer seed coat or inner seed coat and cut the seed, after soaking the seed in strong acid or strong alkali,
Alternatively, a method in which the outer seed coat or the inner seed coat is physically damaged and subsequently placed on the floor.

(4) In the aspect of (1) or (2) above, a method of placing a seed classified as the embryo immature seed is
At least one of the methods described in (2-A) to (2-I) below is selected. (2-A): A method in which the endosperm tissue or the endosperm tissue and the cotyledon tissue are cut so as to include the embryo or the radicle root tissue of the embryo after removing the inner seed coat, and a seed section is placed. (2-B): A method in which the endosperm tissue or the endosperm tissue and the cotyledon tissue are cut so as to include the embryo or the radicle root tissue of the embryo with only the inner seed coat attached thereto. (2-C): A method in which an endosperm tissue or endosperm tissue and cotyledon tissue are cut so as to contain an embryo or an embryo radicle root tissue with an outer seed coat attached thereto. (2-D): after cutting the endosperm tissue or endosperm tissue and cotyledon tissue by any of the methods described in (2-A) to (2-C) above, the embryo or radicle root tissue of the embryo should be further included. A method of implanting a seed section obtained by cutting the side surface of the endosperm tissue or the endosperm tissue and the cotyledon tissue. (2-E): A method of removing the embryo from the seed and placing it on the bed. (2-F): A method of removing the inner seed coat and then placing the seeds without cutting them. (2-G): A method in which seeds are placed as they are without cutting, with only the inner seed coat attached. (2-H): A method in which seeds are placed as they are without cutting the seeds with the outer seed coat attached. (2-I): When it is difficult to remove the outer seed coat or inner seed coat and cut the seed, after soaking the seed in a strong acid or a strong alkali,
Alternatively, a method in which the outer seed coat or the inner seed coat is physically damaged and subsequently placed on the floor.

(5) In the embodiment of (1) or (2) above, a method of placing a seed classified as the embryo mature seed is
At least one of the methods described in (3-A) to (3-I) below is selected. (3-A): A method in which seed seeds obtained by cutting the cotyledon tissue so as to contain the radicle tissue of the embryo are placed after removing the inner seed coat. (3-B): A method in which a seed section obtained by cutting the cotyledon tissue so as to contain the radicle tissue of the embryo is placed with only the inner seed coat attached. (3-C): A method in which a seed section obtained by cutting the cotyledon tissue so as to contain the embryo radicle tissue is placed with the outer seed coat attached. (3-D): Seed section obtained by cutting the cotyledon tissue by any of the methods (3-A) to (3-C) above, and further cutting the side surface of the cotyledon tissue so as to include the radicle tissue of the embryo How to place the floor. (3-E): A method of removing the inner seed coat and then directly placing the seed without cutting. (3-F): A method in which seeds are placed as they are without cutting, with only the inner seed coat attached. (3-G): A method in which seeds are placed as they are without cutting the seeds with the outer seed coat attached. (3-H): In the case of an individual rooted immediately after seeds set and dropped or an individual rooted easily during storage, an already rooted root does not root from a root. Individuals are excised from the tip of radicle tissue and then placed on a germination bed containing auxin. (3-I): When it is difficult to remove the outer seed coat or inner seed coat and cut the seed, after immersing the seed in a strong acid or a strong alkali,
Alternatively, a method in which the outer seed coat or the inner seed coat is physically damaged and subsequently placed on the floor.

(6) In the aspect of (1) or (2) above, at least one of the following (4-A) to (4-D) is a method for placing a seed classified as the hard seed type seed. Is selected from among the methods. (4-A): If the seed is a hard seed type but it is not difficult to remove outer seed coat or inner seed coat or cut off the seed, and if it can be structurally classified as an immature seed of complete embryo, then it is described in (3) above. The method of implanting by the method of any one of (1-A) to (1-E) in the immature seeds of the complete embryo. (4-B): If the seed is a hard seed type but it is not difficult to remove the outer seed coat or inner seed coat or cut the seed, and the seed can be structurally classified as an immature seed, the above-mentioned (4) A method of implanting an embryo immature seed by any of the methods (2-A) to (2-G). (4-C): The embryo described in (5) above , which is a solid seed but is not difficult to remove outer seed coat or inner seed coat or cut off the seed and can be structurally classified as an embryo mature seed. A method of placing a mature seed by any one of the methods (3-A) to (3-F). (4-D): When it is difficult to remove the outer seed coat or inner seed coat and cut the seed, after soaking the seed in any one of strong acid, strong alkali, and boiling water, or physically removing the outer seed coat or inner seed coat How to put the floor after scratching.

(7) In any one of the above (1) to (6) , the processed seeds are washed with water prior to placing them.

(8) In any one of the above (1) to (7) , the treatment method of the seed may be physiological
Sleepy seeds, seeds with slow embryo growth, or ectodermal coats
And seeds that have difficulty removing inner seed coat or cutting seeds
And, after adjusting the solution or mixed solution of cytokinin or gibberellin concentration and mixing ratio suitable for the seeds, which of claims 1-7, characterized in that the plated from in addition to the germination test bed An early method for assaying seeds of woody plants according to any one.

(9) In any one of the above (1) to (7) , the method for treating the seed may be an embryo mature type.
Seeds or solid seeds that share this pattern and
Removal of sleepy seeds or outer and inner seed coats or
For a seed that is difficult to cut, a solution of thiourea or potassium nitrate or a mixed solution thereof is adjusted to a concentration and a mixing ratio suitable for the seed, and this is added to the germination test bed and then placed.

(10) In any one of the above (1) to (7) , the method for treating the seed may be physiological.
Dormant seeds, seeds that need to promote embryonic growth,
Or difficult to remove outer seed coat and inner seed coat or cut seed
The germination test bed is placed under high oxygen partial pressure for fresh seeds .

(11) In any one of the above aspects (1) to (7) , the method for treating the seed may be physiological.
Dormant seeds, seeds that need to promote embryonic growth,
Or difficult to remove outer seed coat and inner seed coat or cut seed
The seed germination test bed is placed under ethylene gas, a mixed gas in which carbon dioxide is added to ethylene gas, or a mixed gas in which carbon dioxide and oxygen are added to ethylene gas.

(12) In any one of the above (1) to (11) , the pretreatment method for the seed may be a fungus.
Sterilization of the seeds is performed on the seeds that are easily invaded or attached .

(13) In any one of the above aspects (1) to (11) , a water absorption method is used as a method for pretreating the seeds.
For seeds that facilitate seed coat removal or seed cutting by
And soak the seeds overnight in water.

(14) In any one of the above aspects (1) to (11) , a surface treatment may be performed as the seed pretreatment method.
The oil and fat component on the surface of the outer seed coat or inner seed coat of the seed is removed from the seed to which the oil and fat component is attached .

(15) In any one of the above (1) to (14) , when measuring the proportion of individuals having germination ability with respect to the seeds placed, the measurement of any one of the following a) to d) The method is selected. a): A method of measuring the proportion of individuals with elongated radicle tissue or individuals with radicle protruding from seed tissue. b): A method of measuring the proportion of individuals whose radicle tissue is curved in the direction of gravity. c): A method of measuring the proportion of individuals in which the cotyledonary tissue is swelled while being firm, or the proportion of individuals in which the cotyledonary tissue has chlorophyll formed and turned green. d): A method of measuring the proportion of individuals with adventitious roots formed.

(16) In the aspect of any one of (1) to (14) above, in measuring the germination ability of the individual having germination ability with respect to the seeds placed, any one of the following i) to v): Is selected. i): A measuring method of measuring the length of radicles that have grown within a certain period or converting the length of the radicles per day. ii): A measuring method of measuring the weight of the embryo increased within a certain period or converting the weight of the embryo per day. iii): A method for measuring the proportion of individuals with germination ability measured by the method according to any one of claims 16 to 16 within a certain period, which is expressed as a percentage. iv): A measuring method of measuring the length of adventitious roots that have grown within a certain period or converting the length of adventitious roots per day. v): A measuring method for measuring the degree of chlorophyll formation within a certain period or converting the degree of chlorophyll formation per day.

(17) In the method of certifying the quality of seeds of woody plants in the present invention, the quality of seeds is certified by presenting at least one of the following a) or a). A): Proportion of individuals having germination ability and / or germination ability obtained by the early assay method for woody plant seeds according to any one of (1) to ( 16) above. B): The actual germination rate estimated from the proportion of individuals having germination ability or germination ability obtained by the early assay method for woody plant seeds according to any one of (1) to ( 16) above.

(18) In the method for designing the seeding amount of woody plant seeds of the present invention, the unit seeding amount of woody plant seeds in the seeding work is calculated by using at least one of the following a) to c) To do. A): Proportion of individuals with germination ability obtained by the early assay method for woody plant seeds according to any of (1) to ( 16) above. B): The actual germination rate estimated from the proportion of individuals having germination ability or germination ability obtained by the early assay method for woody plant seeds according to any one of (1) to ( 16) above. C): Germination rate presented by the quality certification method described in ( 17) above.

[0040]

The present invention has a certain test condition (lightness around room temperature 23 ° C, which is easy to work and economically manages temperature), regardless of the structure and dormancy (physiological properties) of woody plant seeds. The average germination force or germination rate of those seed populations can be determined in a short period (about 1 week). In the conventional method, the germination rate and germination power of all woody plant seeds are set to 1
There is no versatility that can be confirmed around a week.

In the conventional seed germination test method, seed germination progresses smoothly in accordance with the diurnal change in air temperature in the natural world. Therefore, in the germination test, the temperature changing conditions are set according to each species. Is recommended. However, the temperature fluctuation range and temperature range are various, and in order to respond to them, it is necessary to introduce each type of automatic temperature management type facility, and the investment amount for that is huge and it is easy to spread Not really. The present invention can minimize the energy cost and keeps a constant temperature around room temperature (around 23 ° C (21 to 25
℃)) can be carried out under other conditions.

Among seeds, there are anaerobic seeds that prefer darkness for germination (eg, Akigumi etc.), but the present invention is advantageous in that it can be tested in a bright place where it is easy to work without exception.

The dormancy of tree seeds is extremely complicated in combination with its structure, and the rate of post-ripening depending on the species is significantly different depending on the temperature. Therefore, the period until germination can be confirmed in nature is several. It's been two years since the day. However, with this method, without exception, the quality of seeds is evaluated around one week,
It is possible to confirm whether or not it can be used for greening work, and whether it is necessary to change the design.

Seeds that start to germinate immediately after collection (eg:
Quercus serrata, Quercus serrata, etc.) have already emerged from radicles (substantially the first step of germination) at the time of delivery from seed collectors, and seeds that have died are used for greening work, etc. Judgment of whether or not it can be used is a very important matter in terms of greening (vegetation restoration) and construction cost. In this method, when the hypocotyl tissue still retains the secondary meristem vitality, the characteristic that the lateral root / branch can be differentiated from the edge of the cut surface as an adventitious root,
Auxin, which is commonly used as an adventitious root differentiation inducer in cuttings (preferably artificial auxin (NAA))
Etc.) to open up new possibilities for testing woody plant seeds.

Finally, in order to minimize the cost required to submit the test results, it is assumed that the test method applied to each seed should be as simple as possible and that the cheapest drug should be used. The system was planned.

[0046]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention is a development of the above-mentioned isolated embryo method as a basic principle. The isolated embryo method is
Pretreatment method including cutting of seed tissue including radicle tissue, removal of outer seed coat and inner seed coat, removal of embryo from seed, etc. in order to supply water and oxygen necessary for seed germination It is intended to measure the germination rate and the germination ability in a short period of time, including the bed placement method and the treatment method, but there are various problems as described above in general application to woody plant seeds.

In the germination test, the inventors of the present invention first perform classification based on the physiological and structural characteristics of the target woody plant seeds, and in advance the seeds to be individually selected according to the classified structural characteristics. A treatment method, a bed placement method, and a treatment method were found. Then, applying each method selected for the test wood plant seeds, under continuous light irradiation conditions,
For example, by placing on a germination test bed under a constant temperature condition of 23 to 25 ° C., the proportion of individuals having germination ability (hereinafter, referred to as “ratio of germination ability-bearing individuals”) and / or germination power within 7 days It can be measured in a short period of time,
It was found that the germination rate can be estimated from these values.

Specifically, for example, the germination rate is defined by the proportion of individuals having germination ability, or a relational expression (corresponding to the proportion or germination power statistically obtained in advance and the actual germination rate ( Methods such as estimating the germination rate from the calibration curve) are adopted. Hereinafter, embodiments of the early test method for woody plant seeds, the early quality certification method, and the seeding amount design method according to the present invention will be described.

(1) Early Assay Method for Woody Plant Seeds FIG. 1 is a flow chart schematically showing the procedure of the early assay method for woody plant seeds according to the present invention. The main step of the early test method is the classification step 1 based on the structural characteristics of seeds.
1. Step 12 of selecting and performing a pretreatment method performed before bed placement if necessary, Steps 13 and 15 of selecting and implementing an appropriate bed placement method according to classification, and processing performed during bed placement if necessary The method includes steps 14 and 15 of selecting and implementing a method, a step 16 of measuring the proportion of individuals having germination ability, and a step 17 of measuring germinability.

(1-1) Classification Step by Structural Characteristics of Seed In step 11 of FIG. 1, it is judged by the structural identification of the target woody plant seed which classification the seed belongs to. And perform classification. As mentioned above, the treatment of seeds described in the “International Seed Inspection Regulations” involves practically difficulty in the work itself of removing outer seed coats and inner seed coats, because some seeds have small structural characteristics and small seeds. There is. Further, some seeds can be measured for germination ability in a short period of time by giving a plant hormone, a germination-promoting substance, or the like, cutting seed tissue, and the like, even without performing such a treatment involving difficulty. As a result of intensive studies on these points, the inventors classified at least the following four types of seeds according to the structural characteristics of the seeds, and selected a seed placement method suitable for each classified type. By doing so, it was found that the ratio of individuals having germination ability and / or germination ability can be measured more reasonably and rapidly. Classification 1: Complete embryo immature type seed classification 2: Embryo immature type seed classification 3: Embryonic mature type seed classification 4: Hard seed type seeds The structural characteristics of the 4 classified seeds will be described below.

(1-1-1) Classification 1: Complete embryo immature type seeds In the seeds classified into this type, the embryos inside the seeds (cotyledon tissue and radicle tissue) are complete when the seeds are fruiting and mature. It is a seed of a structure that is in an immature state, basically has a completely undeveloped minimal embryo at the end of the endosperm, and these are covered with outer seed coat and inner seed coat. During germination, immature embryos grow inside seeds depending on the nutrient supplementation from endosperm,
The developed embryo shows a germination pattern in which radicles and cotyledons are extruded by exerting pressure on the outer and inner seed coats.

(1-1-2) Classification 2: immature embryo type seeds In the seeds classified into this type, the embryos (cotyledon tissue and radicle tissue) inside the seeds are in an immature state at the stage when the seeds have set and matured. However, compared with the immature seeds of the full embryo, the embryo is well developed. This embryo is a seed having a structure that is located in the endosperm and is covered with outer seed coat and inner seed coat. This seed embryo is
The embryo is more developed than the immature seed embryo, and the cotyledon tissue and radicle tissue are enlarged in the endosperm. In the process of germination,
Underdeveloped embryos grow inside seeds depending on nutrient supplementation from endosperm, and the developed embryos show a germination pattern of protruding radicles and cotyledons by exerting pressure on the outer and inner seed coats.

(1-1-3) Classification 3: Embryo Maturation Type Seeds The seeds classified into this type are mature embryos (radicle root tissue + cotyledon tissue) without endosperm at the stage when the seeds have set and matured. ) Are the only seeds that have a structure in which these are covered by the outer and inner seed coats. These seeds elongate radicles and hypocotyls during germination, develop cotyledon tissues in the aboveground, and further extend epicotyls to develop the first true leaf or keep the cotyledon tissue underground. Shows a germination pattern in which the epicotyl is extended and the first true leaf is expanded in the aboveground part.

(1-1-4) Classification 4: Hard-seed type seeds Seeds classified into this type have the above-mentioned classifications 1 to 3 inside the seeds.
The seeds have the same structure as the seeds classified into the type described in 3, but the outer seed coat and inner seed coat are firm, and the seed tissues have a structure that prevents the supply of oxygen, water, and the like.

(1-2) Selection and Implementation Steps of Pretreatment Method In step 12 of FIG. 1, based on the seed classification performed in step 11, woody plant seeds requiring pretreatment before placement are selected. The appropriate pretreatment method is selected and implemented. Not performed on woody plant seeds that do not require pretreatment.

(1-2-1) Sterilization treatment and water washing For any type of seeds, mold spores and bacteria cannot be removed by ordinary water washing, and against tree species which may be adversely affected during the test. In this case, as a pretreatment, sterilization treatment with sodium hypochlorite, etc. and subsequent water washing are performed. In the germination test, mold development is a major obstacle to accurate measurement of germination rate and germination force. In order to prevent mold, for example, the seeds are sterilized with 2% sodium hypochlorite for 15 minutes, and then the seedlings are placed on a germination test bed by performing an appropriate treatment method described below. Can handle. As a structural characteristic of the seed, this method has a large germination hole like a rabbit seed, and a fungus easily penetrates into the seed, a seed with a flesh fiber firmly attached to the outer seed coat such as bayberry seed, and Tobera. It is desirable to apply it to seeds, etc., which secrete an adhesive substance from the inner seed coat and in which fungi easily attach to the seed surface.

(1-2-2) Immersion in water Seeds having a hard outer seed coat but a soft seed tissue, such as porcupine seeds, nullde seeds, etc., actually have a hard outer seed coat. The rate of destroying the seed tissue also increases. Therefore, it is possible to soften the outer seed coat by soaking the seeds in water overnight so that the seed tissue is not destroyed during cutting. This method, like maple seeds, becomes soft when the outer seed coat absorbs water when removing the outer seed coat, making it easier to remove the outer seed coat or cutting the seed by absorbing water like the Tobera seed. It may be applied to the case where it becomes easy. This method can be applied not only to the outer seed coat but also to the case where it is difficult to remove and cut the inner seed coat.

(1-2-3) Bleaching or Washing In any type of seeds, in order to promote water absorption and oxygen respiration by the seeds only when the mucus liquid substance or wax-like substance adheres to the seed coat surface, As a pretreatment, bleaching with sodium hypochlorite or washing with neutral detergent is performed. For example, there are those in which oil and fat components are attached to the surface of seeds such as red wrinkle seeds. In such seeds, the oil and fat impede the water absorption of the seeds, so that the germination rate becomes slow. For these seeds, the germination rate can be increased by, for example, thoroughly washing the oil and fat components on the surface of the outer seed coat or the inner seed coat of the seed with an appropriate amount of a neutral detergent or the like and then placing the seeds on the seeds.

(1-3) Step of selecting a bed placement method In step 13 of FIG. 1, a seed bed placement method is selected based on the seed classification performed in step 11.
There are several possible placement methods for each seed type and the appropriate placement method is selected. The bed placement method includes physically processing the seed, such as cutting or cutting the seed.

(1-3-1) Implanting Method for Completely Immature Type Seeds Germination of seeds classified as immature type complete embryos occurs even if germination conditions such as temperature and moisture are uniform. It takes more time to germinate than seeds of other structures only for the time to develop. Therefore, the following bed placement methods (1-A) to (1-F) are applied to this type of seed. (1-A): After removing even the inner seed coat, a seed section obtained by cutting the endosperm tissue so as to contain the embryo is placed. (1-B): A seed section obtained by cutting the endosperm tissue so as to contain the embryo is placed with only the inner seed coat attached. (1-C): A seed section obtained by cutting the endosperm tissue so as to contain the embryo is placed with the outer seed coat attached. (1-D): After cutting the endosperm tissue by any one of the methods (1-A) to (1-C) above, the side surface of the endosperm tissue is cut so as to further contain the embryo, and the seed slice is placed. (1-E): The embryo is taken out from the seed and placed on the bed. (1-F): When it is difficult to remove the outer seed coat or inner seed coat and cut the seed, after soaking the seed in strong acid or strong alkali,
Alternatively, the outer seed coat or the inner seed coat is physically damaged and then placed on the floor.

Description of implantation methods (1-A) to (1-C) By implanting seed slices obtained by cutting the endosperm tissue so that the embryos are always included, the supply of oxygen, water, etc. is increased and the growth of embryos is increased. Can be urged. Also, the reasons for cutting this seed are:
By cutting the endosperm tissue and making the seed slice smaller,
By that amount, the embryo developed in the endosperm tissue can be pushed out of the seed section earlier, and the degree of development of the embryo can be confirmed in a short period of time. From this fact, if the size of the seed fragment is different, the degree of development of the embryo cannot be accurately confirmed. Therefore, the size of the seed fragment should be made uniform as much as possible. In these bed placement methods, it is basically desirable to remove outer seed coat and inner seed coat as much as possible in order to enhance the supply of oxygen, water and the like. However, if it is difficult to remove the outer seed coat and the inner seed coat depending on the size and shape of the seed, it is not necessary to dare to do so. Examples of tree species using these bed placement methods include, for example, Aoki (combination with bed placement method (1-D)), bed rattle, etc. for bed placement method (1-A), and boxwood, tobera, nanten for bed placement method (1-B). Etc., such as the flooring method (1-C), there is a sea buckthorn.

Description of bed placement method (1-D) When the seed is large, after applying the seed cutting method described in the above bed placement methods (1-A) to (1-C), oxygen, water, etc. In order to further increase the supply of seeds and to allow the plant hormones and germination promoters added to the germination bed to permeate the embryo early, the side of the endosperm tissue is cut to include the embryo, and the seed section containing the embryo is blocked. By placing it in the shape of
It is possible to increase the germination rate. Tree species that use this flooring method include, for example, Aoki (combined with the flooring method (1-A)).

Description of Implantation Method (1-E) When the growth of embryos is extremely slow, it is possible to take out only the embryos from the seeds and immobilize them.

Description of bed placement method (1-F) In this type of seed, when the seed size is small and removal of outer seed coat and inner seed coat or cutting of seed tissue is difficult, for example, strong acid If the seed is soaked with sodium hypochlorite, etc. if it is a strong alkali such as sulfuric acid, the seed coat can be thinned, or the seed surface can be physically scratched using a rice mill or juicer mixer to cut the seed. Leave it as it is without doing it. As a result, the germination rate can be increased by promoting the absorption of water, plant hormones and oxygen.

(1-3-2) Implanting method for immature embryo seeds Germination of seeds classified as immature embryo seeds is carried out even if germination conditions such as temperature and water are the same as in the case of immature embryo seeds. The time until the embryo develops inside the seed requires more time to germinate than other types of seed.This time is longer for smaller embryos and larger for embryos. It will be a short period. Therefore, this type of seed has the following (2-A)
Apply the floor placement method as in (2-I). (2-A): After removing the inner seed coat, a seed section obtained by cutting the endosperm tissue or the endosperm tissue and the cotyledon tissue so as to include the embryo or the radicle root tissue of the embryo is placed. (2-B): A seed section obtained by cutting the endosperm tissue or endosperm tissue and cotyledon tissue so as to include the embryo or the radicle of the embryo is placed with only the inner seed coat attached. (2-C): With the outer seed coat attached, a seed section obtained by cutting the endosperm tissue or the endosperm tissue and the cotyledon tissue so as to include the embryo or the radicle root tissue of the embryo is placed. (2-D): The endosperm tissue or endosperm tissue and cotyledon tissue are cleaved by any of the methods described in (2-A) to (2-C) above, and the endosperm tissue or endosperm is further formed so as to further contain an embryo. A seed section obtained by cutting the side surface of the tissue and the cotyledon tissue is placed. (2-E): Remove the embryo from the seed and place it in the bed. (2-F): After removing the inner seed coat, the seeds are directly placed on the bed without cutting. (2-G): Place the seed without cutting the seed without cutting the inner seed coat. (2-H): The seeds are left on the bed without cutting, with the outer seed coat attached. (2-I): When it is difficult to remove the outer seed coat or inner seed coat and cut the seed, after soaking the seed in a strong acid or a strong alkali,
Alternatively, the outer seed coat or the inner seed coat is physically damaged and then placed on the floor.

-Explanation of implantation methods (2-A) to (2-C) Oxygen or water is obtained by implanting seed endosperm tissue or seed slices obtained by cutting endosperm tissue and cotyledon tissue so as to contain embryonic radicle tissue. It is possible to promote the growth of the embryo by increasing the supply of such as. The reason for the cutting of this seed tissue is that cutting the endosperm tissue to reduce the size of the seed slice causes the embryo developed in the endosperm tissue to protrude to the outside of the seed slice by that much, which leads to the development of the embryo. Sometimes, it is possible to confirm the degree of in a short period of time. From this fact, if the size of the seed fragment is different, the degree of development of the embryo cannot be accurately confirmed. Therefore, the size of the seed fragment should be made uniform as much as possible. In these bed placing methods, it is basically desirable to remove outer seed coat and inner seed coat as much as possible in order to enhance the supply of oxygen, water and the like. However, if it is difficult to remove the outer seed coat and the inner seed coat depending on the size and shape of the seed, it is not necessary to dare to do so. As a tree species using these bed placement methods, for example, bed placement methods (2-B) include privet, echinacea, natsuhaze, dogwood, masaki, pokeweed, camellia, Himeshara, Hordeum, etc., such as bed placement methods (2-C), There are yellowfin, trevana, hashidoi, murasakihasidoi, and the like.

Description of bed placement method (2-D) When the seed is large, after applying the seed cutting method described in the bed placement methods (2-A) to (2-C) described above, oxygen, water, etc. Endosperm tissue or endosperm tissue and cotyledon tissue so as to include embryos or radicles of embryos so that plant hormones or germination promoters added to germination beds can spread early into the radicle tissue. The germination rate can be increased by cutting the side surface of the seed and placing the seed pieces containing the embryo or embryo radicle tissue in a block shape.

-Explanation of the implantation method (2-E) When the growth of the embryo is extremely slow, it is possible to take out only the embryo from the seed and place it. Examples of tree species that use this bed placement method include mouse peach.

Description of bed placement methods (2-F) to (2-H) Cutting of seeds when the development of the embryo is advanced or when the growth of the embryo is fast, or when plant hormones or germination promoting substances are added. If you germinate in a short period of time without performing
When it is difficult to cut the seeds because the seeds are small, the seeds are not cut and placed as they are. In these bed placing methods, it is basically desirable to remove outer seed coat and inner seed coat as much as possible in order to enhance the supply of oxygen, water and the like. However, if it is difficult to remove the outer seed coat and the inner seed coat depending on the size and shape of the seed, it is not necessary to dare to do so. Examples of tree species that use these bed placement methods include wolfberry, gardenia, sakaki and the like as bed placement methods (2-G).

Description of bed placement method (2-I) In this type of seed, when the seed size is small and it is difficult to remove the outer seed coat and inner seed coat or cut the seed tissue, for example, strong acid is concentrated. If the seed is soaked with sodium hypochlorite, etc. if it is a strong alkali such as sulfuric acid, the seed coat can be thinned, or the seed surface can be physically scratched using a rice mill or juicer mixer to cut the seed. Leave it as it is without doing it. As a result, the germination rate can be increased by promoting the absorption of water, plant hormones and oxygen. For example, the floor placement method (2-I) includes "Igiri".

(1-3-3) Implantation Method for Embryo Matured Seeds The seeds classified as embryo matured seeds are subjected to the following implantation methods (3-A) to (3-I). (3-A): After removing even the inner seed coat, a seed section obtained by cutting the cotyledon tissue so as to contain the embryo radicle tissue is placed. (3-B): A seed section obtained by cutting the cotyledon tissue so as to include the embryo radicle tissue is placed with only the inner seed coat attached. (3-C): With the outer seed coat attached, a seed section obtained by cutting the endosperm tissue and cotyledon tissue so as to include the radicle tissue of the embryo is placed. (3-D): After cutting the cotyledon tissue by any of the methods (3-A) to (3-C), the side surface of the cotyledon tissue was further cut so as to include the tip of the radicle root tissue of the embryo Place seed slices. (3-E): After removing the inner seed coat, the seeds are placed as they are without cutting. (3-F): With the inner seed coat only, the seeds are placed as they are without cutting. (3-G): With the outer seed coat attached, the seeds are placed as they are without cutting. (3-H): In the case of an individual rooted immediately after the seeds set and dropped, or an individual rooted easily in storage, an already rooted root does not root the radicle. After excising the tip of the radicle tissue, the individual is placed on the germination bed containing auxin. (3-I): When it is difficult to remove the outer seed coat or inner seed coat and cut the seed, the seed is soaked in a strong acid or a strong alkali, or after the outer seed coat or the inner seed coat is physically damaged, it is post-bed. Method.

-Explanation of implantation methods (3-A) to (3-C) By implanting seed slices obtained by cutting the cotyledon tissue so as to include the radicle tissue of the embryo, the supply of oxygen, water, etc. is increased to promote the growth of the embryo. It is possible to promote the growth of root tissue. In these bed placing methods, it is basically desirable to remove outer seed coat and inner seed coat as much as possible in order to enhance the supply of oxygen, water and the like. However, if it is difficult to remove the outer seed coat and the inner seed coat depending on the size and shape of the seed, it is not necessary to dare to do so. Examples of tree species that use these bed placement methods include, for example, the bed placement method (3-A), such as Sharimbai, Camellia japonica, Sudazii, and the bed placement method (3-A), and those used in combination with the bed placement method (3-D). Is Arakashi, Ubamegashi, Chestnut, White Oak,
Flooring method (3-B), such as tea tree, Akigumi, Oyamazakura, Uwamizuzakura, Sasanqua, Rowan, Fuyo, Moccok, Yamazakura, bayberry, etc.
B) is used in combination with the bed placement method (3-D), and the bed placement method (3-C) includes halibut, beech, and Neubara.

Description of bed placement method (3-D) When the seeds are large, after the seed cutting method described in the above bed placement methods (3-A) to (3-C) is taken, oxygen and water are added. In order to further increase the supply of plant germs, etc., the side of the cotyledonary tissue is cut to include the embryonic radicle tissue, so that the plant hormones and germination promoters added to the germination bed may permeate into the radicle tissue early, The germination rate can be increased by placing the seed slice containing the root tissue in a block shape. Examples of tree species that use this flooring method include, for example, those that use the flooring method (3-A) in combination, such as Arakashi, Ubamegashi, Chestnut, Shirokashi, Camellia, etc.
The ones that use the above-mentioned method include matebashii, and the ones that use the bed placement method (3-H) include Abemaki and Kunugi.

Description of bed placement methods (3-E) to (3-G): Does not have physiological diapause, and germinates in a short period of time without cutting seeds, and phytohormones and germination-promoting substances By adding, etc., seeds that germinate in a short period of time without cutting the seeds, or have difficulty in cutting the seeds because the seeds are small, or because cotyledon tissue constitutes the seed tissue intricately, In the case of seeds whose seed tissue becomes fine fragments when cutting is performed, the seeds are not cut and placed as they are. Here, the seed in which the cotyledon tissue is intricately forming a seed tissue is a seed in which the cotyledon tissue is spirally formed inside the outer seed coat (the same applies hereinafter). In these bed placing methods, it is basically desirable to remove outer seed coat and inner seed coat as much as possible in order to enhance the supply of oxygen, water and the like. However, if it is difficult to remove the outer seed coat and the inner seed coat depending on the size and shape of the seed, it is not necessary to dare to do so. As tree species using these bed placement methods, for example, in the bed placement method (3-E)
Flooring method (3-F) such as tea tree, yam camellia, etc. can be used as a maple (3F)
-G) includes Achillet, wig, zelkova, crape myrtle, birch, birch, yashabushi, yamahanoki and others. In addition, as for the Sharimbai, which is the seed to which the above-mentioned bed placement method is applied, tea tree, Camellia japonica, normally,
It is advisable to carry out the test using the bed placement method (3-A) that results in a faster germination rate.

Regarding the bed placement method (3-H), in the case of individuals such as oaks that root immediately after seed set and fall, or individuals that easily root during storage, they have already rooted. Individuals are treated by cutting radicles from their roots and individuals not rooting by cutting off the tip of radicle tissue and placing it on a germination bed containing NAA and the like. Examples of tree species using this bed placement method include Abemaki and Kunugi (bed placement method (3-
D) together), Quercus, Quercus etc.

Bedbed method (3-I) With this type of seed, when the seed size is small and it is difficult to remove the outer seed coat and inner seed coat and to cut the seed tissue, for example, strong acid is concentrated sulfuric acid. If the alkali is strong, the seed coat can be thinned by immersing the seed in sodium hypochlorite, etc., and the seed can be cut by physically scratching the seed surface using a rice polishing machine, juicer mixer, etc. Instead, leave the floor as it is. As a result, the germination rate can be increased by promoting the absorption of water, plant hormones and oxygen.

(1-3-4) Implanting method of hard-seed seeds To promote germination of seeds classified as hard-seed seeds, solid outer seed coat and inner seed coat are removed, partially removed, damaged, and thinned. The first condition is to promote the supply of oxygen, water, etc. Therefore, for this type of seed, the following bed placement methods (4-A) to (4-D) are applied. (4-A): If the seeds are hard seeds but the removal of outer or inner seed coats or cutting of the seeds is not difficult, and the seeds can be structurally classified as immature seeds of complete embryos, the above-mentioned full cup immature seeds Place the bed by any one of (1-A) to (1-E). (4-B): It is a hard seed type but it is not difficult to remove the outer seed coat or inner seed coat or cut the seed, and if the seed can be structurally classified as an immature seed, in the aforementioned embryo immature seed ( 2
-Place one of the methods from (A) to (2-G). (4-C): It is a hard seed type but it is not difficult to remove outer seed coat or inner seed coat or cut the seed, and if it can be structurally classified as an embryo mature seed, in the above-mentioned embryo mature seed ( 3
-Place one of the methods from (A) to (3-F). (4-D): If it is difficult to remove the outer seed coat or inner seed coat and cut the seed, after soaking the seed in strong acid, strong alkali, or boiling water, or by physically removing the outer seed coat or inner seed coat Place it on the floor after injuring it.

-Explanation of bed placement methods (4-A) to (4-C) In consideration of the size and workability of seeds, bed placement methods (1-A) to (1-E in the case of fully embryonic immature seeds ), A bed placement method (2-A) to (2-G) for immature embryo seeds, and a bed placement method (3 for embryo mature seeds).
-A) to (3-F) can be dealt with by applying a suitable bed placement method by any of the methods described in each. The tree species using these bed placement methods include, for example, bed maturation seed embryo placement methods (3-A) such as Nankin goby, and embryo maturation type seed placement methods (3-B) weasel, komatsuna, sardines, There are alder trees, yamahagi, etc., and the embryo-mature seed seed placement methods (3-C) include Nurude, Yamaurushi, and Yamahase, and the embryo-mature seed seed placement methods (3-F) include Akamega wrinkles.

Regarding the bed placement method (4-D), since the seeds are small, the complete embryo immature type seed, embryo immature type seed, and embryo mature type described in the bed placement methods (4-A) to (4-C) described above. If the operation of each seed bed method is not possible, or if it is difficult to remove the outer and inner seed coats or cut the seeds, for example, concentrated sulfuric acid in the case of strong acid, hypochlorous acid in the case of strong alkali By dipping the outer or inner seed coat by dipping it in sodium acid or the like, softening the outer or inner seed coat by dipping it in hot water, or by physically injuring part of the outer or inner seed coat and placing it on the bed. Can handle. Tree species using this method include, for example, enoki.

(1-4) Selection Step of Treatment Method In step 14 of FIG. 1, a seed treatment method is selected. The method for treating woody plant seeds in the present invention,
It refers to a method of treatment for imparting predetermined environmental conditions to woody plant seeds during the time period of planting woody plant seeds. Even if placed by any of the above placement methods depending on the structural characteristics of each seed, the physiological characteristics of the seed such as the physiological dormancy of the seed, the growth rate of the embryo, or the outer seed coat and the inner seed coat. Some seeds may not germinate within 7 days due to structural properties that are extremely difficult to remove and cut. Accurate values cannot be measured for seeds of tree species that do not germinate within 7 days. However, even with such seeds, it is possible to accelerate the germination rate by performing the germination-promoting treatment in parallel during the placement and to measure the accurate value within 7 days.

(1-4-1) Treatment with Growth-Promoting Solution When the seed has physiological dormancy, when the growth of the embryo is slow, removal of outer and inner seed coats or cutting of seeds is possible due to the structural characteristics of the seed. If it is difficult, cytokinin (preferably benzyladenine etc. (BA)), gibberellin (G
A), a solution of thiourea, potassium nitrate, or a mixed solution thereof is adjusted to a concentration and a mixing ratio suitable for the seeds, and then this is added to the germination test bed and then placed, thereby shortening the inspection period. be able to. Specifically, it is advisable to impregnate the respective solutions into a germination bed such as a filter paper.

BA, which is a representative of cytokinins, and GA, which is used for inducing activation of hydrolase group, are plant hormones, which improve the reducing power (NADPH) productivity and thereby induce the cotyledon-developing power. can get. BA and GA promote the growth of radicles, hypocotyls, and cotyledons, and thus can be applied to all seeds. The concentrations of BA and GA vary depending on the intensity of physiological dormancy, the growth rate of embryos, the size of seeds, and the like. Basically, the higher the physiological dormancy, the slower the growth of embryos, and the larger the seed size, the higher the concentration. Also, if the dormancy is stronger, B
It is effective to use a mixed solution of A and GA.

In the case of embryo matured seeds or solid seeds sharing this type, if the seeds have physiological diapause, or if the seeds have structural characteristics, the outer and inner seed coats can be removed or the seeds can be cut. If it is difficult, after adjusting the concentration of a solution of thiourea and potassium nitrate, which are germination promoting substances, to a concentration suitable for the seeds, adding this to the germination test bed and then placing it on the bed can shorten the inspection period. It will be possible. Specifically, it is advisable to impregnate the respective solutions into a germination bed such as a filter paper.

Thiourea as a diapause breaker promotes only the growth of radicles and hypocotyls, and has a small effect of promoting the growth of cotyledons. It does not promote the growth after germination, but it depends on the species. Has a dormancy breaking effect. Therefore, like seeds of maples, the growth of cotyledons is almost complete, and the seeds are germinated only by the growth of radicles and hypocotyls. The concentration of thiourea varies depending on the strength of dormancy, seed size, etc. Basically, when the dormancy is strong and the seed size is larger, higher concentration is required. BA and G
Although A has the same effect, these substances are expensive in price. On the other hand, since thiourea is inexpensive, it is possible to perform the test at low cost by using it for seeds that can use thiourea due to its physiological characteristics.

Potassium nitrate as the electron acceptor and amino acid raw material promotes germination by promoting respiration of seeds and supplementing biosynthesis of amino acids and proteins. Small seeds such as Betula birch seeds contain amino acids,
Since the storage amount of protein is small, it becomes possible to measure the germination force within a short period of 7 days by giving potassium nitrate. This method is effective in adding potassium nitrate alone, and further adding it to seeds that do not germinate in a short period of time with only plant hormones or thiourea. The concentration of potassium nitrate is the strength of each dormancy,
It depends on the size of seeds. Basically, when the dormancy is strong and the seed size is larger, higher concentration is required.

(1-4-2) Treatment by High Oxygen Partial Pressure When the seed has physiological dormancy, when it is necessary to promote the growth of the embryo, the removal of outer seed coat and inner seed coat depending on the structural characteristics of the seed, If it is difficult to cut the seeds, it can be handled by placing the germination test bed under high oxygen partial pressure as another method. Since the seeds breathe when they germinate to produce the energy required for germination, it is possible to enhance the respiratory activity of the seeds and increase the germination rate by placing the seeds under conditions of high oxygen concentration. become.

Specifically, the petri dish on which the seeds have been placed is placed in a container, such as a desiccator, which can be closed and decompressed, and the gas in the container is replaced with oxygen to be placed on the bed. Further, in the case of seeds having a strong dormancy or the growth of embryos is very slow, the germination rate can be increased by combining with the above method (1-4-1).

(1-4-3) Treatment with Growth Promoting Gas When the seed has physiological dormancy, it is necessary to promote the growth of the embryo. If it is difficult to cut, the germination test bed is ethylene gas, a mixed gas of carbon dioxide added to ethylene gas, or a mixed gas of carbon dioxide and oxygen added to ethylene gas, as another method. The germination period can be shortened by placing it below.
The addition of ethylene gas enhances the respiratory activity of seeds and promotes germination. By adding carbon dioxide to it, the effect of ethylene gas is increased by supplementing the necessary carbon skeleton, and by adding oxygen, the respiratory activity effect of ethylene gas is strengthened and it is possible to further accelerate the germination rate. Become.

Specifically, the petri dish on which the seeds have been placed is placed in a container that can be sealed, such as a desiccator, and ethylene gas of a specified concentration is added to the container and placed. The concentration of ethylene gas varies depending on the strength of seed dormancy, the growth rate of embryos, the size of seeds, and the like.
Further, germination rate can be increased by adding together with ethylene gas a carbon dioxide that provides a carbon skeleton necessary for seed growth or a mixed gas in which oxygen necessary for respiration is further mixed. It should be noted that, in the case where seeds with strong dormancy or the growth of embryos are very slow, etc., the germination rate can be increased by combining with the method of (1-4-1) above.

(1-5) Steps for carrying out the bed placement method and treatment method In step 15 of FIG.
The bed placement method and treatment method are carried out according to the bed placement method and treatment method selected in 4, and the individual to be placed in bed is thoroughly washed with water before placement. Since the work of step 15 involves the work of removing the pericarp and cutting the seeds, it is possible to suppress the generation of mold and the like by performing a cleansing before the bed placement. In particular, it is important to remove the substances secreted from the seeds by cutting with washing in order to suppress the generation of mold. In carrying out the method of placing and treating, the target woody plant seeds are placed in a Petri dish of a diameter suitable for the size of each seed, and a moistened filter paper is applied to the germination floor to give a temperature of 23 ° C. The standard method is to place it under the floor.
It should be noted that a container other than a petri dish or a germination bed made of another material may be used. Although the temperature conditions are not limited, the assay method of the present invention is characterized in that the assay can be completed at a constant temperature near room temperature.

(1-6) Measuring Step of Ratio of Germinating Ability Retaining Individuals In step 16 of FIG. 1, the ratio of seeds retaining a germinative ability of the seeds subjected to the above-mentioned bed placement method and treatment method is measured. As a specific measuring method, any of the following a) to d) is selected and applied according to the tree species as a criterion for judging that the seed has germination ability. Then, the "numerical value" of the ratio obtained as a result of the measurement is defined as the "proportion of individuals having germination ability".

(1-6-1) Measuring Method a) The proportion of individuals with elongated radicle tissue or individuals with radicle protruding from seed tissue is measured.

(1-6-2) Measuring method b) The proportion of individuals whose radicle tissue is bent in the direction of gravity is measured.

(1-6-3) Measuring Method c) The proportion of individuals in which the cotyledonary tissue is swelled while being firm, or the proportion of individuals in which chlorophyll is formed in the cotyledonary tissue and turned green is measured.

(1-6-4) Measuring method d) The percentage of individuals with adventitious roots formed is measured.

(1-7) Step of measuring germination ability In step 17 of FIG. 1, the germination ability of seeds subjected to the above-mentioned bed placing method and treatment method is measured. As a specific measuring method, any one of the following i) to v) is selected and applied . Then, the “numerical value” obtained as a result of the measurement is defined as “germination power”.

(1-7-1) Measuring method i) Length of radicles that have grown or breached seed tissue from within the seed tissue for a certain period (preferably within 7 days) or this value is converted per day The extended length is referred to as "numerical value".

(1-7-2) Measuring method ii) The weight of the embryo before being placed is measured, and the weight of the embryo is measured again within a certain period (preferably within 7 days) to calculate the calculated embryo. The weight increase rate of or the weight increase rate obtained by converting this value per day is referred to as a "numerical value".

(1-7-3) Measurement method iii) Growth within a certain period (preferably within 7 days) by any of the above-mentioned measurement methods a) to d) of the proportion of individuals having germination ability. The percentage of individuals showing growth was defined as the germination force, and the percentage of individuals showing growth was taken as the "numerical value".

(1-7-4) Measuring method iv) The length of adventitious roots grown within a certain period (preferably within 7 days) or the length of the grown root obtained by converting this value per day is referred to as "numerical value". To do.

(1-7-5) Measuring Method v) The degree of chlorophyll formation was measured within a certain period (preferably within 7 days) or the value obtained by converting the degree of chlorophyll formation per day was calculated as " Numerical value ". As a method for measuring the degree of chlorophyll formation, a known method such as a chlorophyll meter (colorimeter) can be used.

(2) Method for Proving Early Quality of Woody Plant Seeds FIG. 2 is a flow chart schematically showing the procedure of the method for proving early quality of woody plant seeds according to the present invention. The main steps of the quality certification method include two steps, that is, a calibration curve generation step 20 and a germination rate estimation and quality certification step 30.

(2-1) Step for Creating Calibration Curve Step 20 in FIG. 2 is a step for creating a calibration curve.
This step 20 is a step of preparing in advance a calibration curve necessary for carrying out the next germination rate estimation and quality certification step 30. The calibration curve is a graph showing the correlation between (the numerical value of) the germination force obtained by performing the early test method for woody plant seeds and the actual germination rate obtained after actually sowing the seeds. is there. Also, a graph showing the correlation between the measurement result (numerical value) of the ratio of individuals having germination ability and the actual germination rate is included. In step 21 of FIG. 2, the above-described early test method is performed to obtain the ratio of the individuals having germination ability or germination ability as a numerical value. In parallel with this, in step 22, the actual germination rate when the seed is actually sown is measured. Then, in step 23, the correlation derived between the germination force and the actual germination rate is represented in a graph, and a calibration curve is created. FIG. 3 is a graph showing an example of the calibration curve created in step 20. This calibration curve shows the correlation between the germination force and the actual germination rate. In addition, it is a preferable example to represent the correlation by a graph as shown in the figure, but it may show a correlation derived between the ratio of germination ability-bearing individuals or germination power and the actual germination rate. For example, the expression format is arbitrary.

(2-2) Germination Rate Estimation and Quality Certification Step After obtaining the calibration curve in advance in step 20, step 30
Perform germination rate estimation and quality verification steps of. In step 31, the above-described early test method is performed on the woody plant seed that is the target of the quality certification, and the ratio of individuals having germination ability and / or germination ability are obtained as numerical values. Next step 32
In, the numerical value of the germination rate obtained from the calibration curve is estimated as the actual germination rate by applying the calibration curve to the ratio of germination ability-bearing individuals or the germination power obtained (hereinafter, referred to as “estimated germination rate”). ). This method can be easily applied to trees with relatively low physiological dormancy. Then, in step 33, quality certification is performed by the estimated germination rate obtained in step 32, or by the estimated germination rate obtained in step 32 and the proportion and / or germination ability of the individuals having germination ability obtained in step 31. “Providing quality certification” specifically means, for example, expressing the obtained numerical data in the form of a quality certificate, etc., and presenting supplementary explanations such as measurement methods and situation photographs according to the situation. (Hereinafter the same). As a result, it is possible to carry out the quality certification of the woody plant seed in a short period (preferably within 7 days).

(3) Method for designing seeding amount of woody plant seed FIG. 4 is a flow chart schematically showing the procedure of the method for designing seeding amount of woody plant seed according to the present invention. The main steps of the sowing rate design method are a step of measuring the ratio of germination ability-bearing individuals and / or germination ability, a step of estimating germination rate by applying a calibration curve, and a step of germination rate by one or more of the steps of quality verification. And a step of determining the seeding amount using the obtained germination rate.

(3-1) Measurement of Proportion of Germinating Ability Retaining Individuals and Germination Ability In step 41, the above-mentioned early test method is applied to the seeds of the woody plant to be sown, and the germinating ability retaining individuals are And / or germination power as a numerical value.

(3-2) Germination Rate Estimation Step In step 42, the calibration curve obtained in advance as in step 20 of FIG. 2 is used as the ratio and / or germination ability of the individuals having germination ability obtained in step 41. Apply to obtain the estimated germination rate by estimating the actual germination rate.

(3-3) Quality certification step In step 43, the estimated germination rate obtained in step 42, or the estimated germination rate obtained in step 42 and the proportion of individuals having germination ability obtained in step 41 and / Alternatively, the quality is verified by germination.

(3-4) Step of determining seeding amount In step 44, the seeding amount is determined by calculating the unit seeding amount of the seeds of the woody plant when carrying out the seeding work.
The seeding amount is determined by using the ratio of individuals having germination ability obtained in step 41, using the estimated germination rate obtained in step 42, or the germination rate included in the quality certification obtained in step 43. May be used. In any case, the early test method of the present invention can reliably obtain reliable numerical data within a fixed period within 7 days. As a result, it is possible to greatly promote the application of native tree seeds, which had been difficult to use until now, to revegetation, and to enable efficient vegetation restoration using biodiversity using native species. become. In addition to seeds used for revegetation, in the field of seedling production, etc., efficient seedling production can be performed by using this method to test the germination rate.

(4) Example of application of the present invention In the early assay method for woody plant seeds according to the present invention, the treatment method and the like are appropriately selected according to the characteristics of the seeds, and the germination rate is measured within a short period of 7 days or less. This is a method of estimating. In order to effectively utilize this method, it is preferable to effectively combine the above-mentioned pretreatment method, bed placement method, and treatment method. An application example combining specific methods,
It is shown below.

(4-1) Example of application to immature seeds of complete embryo The immature seeds of complete embryo generally undergo a long-term after-ripening process before the germination ability can be expressed. However, some seeds that are actually naturally collected do not always have the same post-ripening stage because of different collection times. But,
As a general rule, seeds of this type are considered not to undergo post-ripening, and the following methods (4-1-1) to (4-1-3) may be applied.

(4-1-1) For example, when the size is about 5 mm or more, the embryo is removed from the endosperm to take out germination ability (plant hormones GA, BA, GA + BA).
And / or oxygen substitution). After that, if necessary, the degree of growth of radicles or cotyledons that have grown around one week is measured using a micrometer or the like, and pigment formation is investigated to determine the proportion (germination rate) of individuals with germination ability and seeds. It is recommended to test the germination power of.

(4-1-2) For example, in the case of a small seed having a diameter of about 4 mm or less, the seed is cut so as to include an embryo,
Care should be taken to locate the embryo within the endosperm of the seed section. It is advisable to place the plant for about 1 week under the above-mentioned plant hormone and / or oxygen substitution, measure the degree of growth of the radicle or cotyledon that has grown, if necessary, using a micrometer or the like, and investigate the degree of pigment formation.

(4-1-3) For example, for seeds to which the embryo extraction method cannot be applied due to its smaller size, the seeds are put in a rice mill or the like and gently stirred to scratch or scratch the surface of the seed coat. In order to promote the invasion of the above plant hormones and oxygen, reduce the physical pressure of embryo development by the seed coat, and allow germination in about one week, and show the signs of germination with a magnifying glass if necessary. Alternatively, it may be detected under a stereoscopic microscope.

(4-2) Example of application to immature embryo seeds In immature embryo seeds, post-ripening of the embryo has progressed to some extent at the time of seed completion, and the period until the embryo matures in the seed is complete. Relatively short compared to immature seeds of the cup. This includes those that have a low degree of dormancy and are already in a dormant awake state at the time of collection. Therefore, a micrometer is not required to test the germination ability of this type of seed, and the following methods (4-2-1) to (4-2-7) may be applied.

(4-2-1) For example, when post-ripening is almost completed and only the embryo can be removed, the degree of radicle elongation on the water-donated filter paper in the atmosphere (per unit time) It is recommended to test the germination force based on the amount of elongation.

(4-2-2) For example, in the case where only the embryo cannot be extracted even after the post-ripening is almost completed and the size of the seed is about 3 mm or more, the portion containing the embryo from the seed In order to isolate only the seeds, the cutting of the seeds or the cutting of the side of the seed section is combined with the cutting of "embryo tissue + endosperm" to facilitate the oxygen respiration of these seed sections under the water-donating atmosphere. By so doing, the growth of the embryo is promoted, and the germination ability may be assayed based on the degree of growth of the protruding radicle or cotyledon or the degree of pigment formation in the cotyledon.

(4-2-3) For example, if the seed size is the same as in the above (4-2
-2) If post-ripening is not completed and dormancy remains, phytohormones (GA alone or a combination of GA and BA) under air in addition to (4-2-2) above It is advisable to administer the drug and conduct a similar study.

(4-2-4) For example, if the size of the seed is the same as in the above (4-2-
2) When the post-ripening is not completed and dormancy remains, and the type of seed that cannot induce embryo development to be seen in about 1 week in (4-2-3) above is It is advisable to carry out oxygen substitution of the atmosphere under the donation.

(4-2-5) For example, if the size of the seed is the same as in the above (4-2-
2) When post-ripening is not completed and dormancy remains, the development of embryos to be seen in about 1 week also in (4-2-3) and (4-2-4) above. See (4
-2-3) and (4-2-4) above may be combined, and the seed section may be placed together with the plant hormone under oxygen substitution in the atmosphere.

(4-2-6) For example, in the case where the size of the seed is 2 mm or less, which is so small that it is difficult to cut, the seed that has been matured and does not germinate in about 1 week under a water-supplied atmosphere In the case of the seed, without cutting the seed, the (4-2-3),
It is advisable to apply either (4-2-4) or (4-2-5) above to test the germination rate or germination power.

(4-2-7) For example, in the case of seeds of the type in which the sign of germination cannot be detected even after about 1 week even by the above (4-2-6), the seeds are put in a rice polishing machine or the like and stirred. The seed surface is scratched to reduce the physical pressure exerted by the seed coat on the growth of the embryo, accelerate the germination rate by promoting the absorption of plant hormones and oxygen, and the germination rate and germination ability may be tested.

(4-3) Example of Application to Embryo Matured Seed In the embryo matured seed, the embryo has completely developed morphologically at the time of seed completion, and the endosperm tissue in the seed disappears. However, even the post-ripening of the embryo itself is not completely completed. The test method for these seeds should be divided into the following three groups (4-3-1) to (4-3-3) based on the growth pattern.

(4-3-1) Non-diapause (Example of application to a seed group when the post-ripening is incomplete requires some time before germination) Example 1): It is advisable to place seeds of the type in which radicles are projected within about 1 week under normal water-supplying atmospheric conditions, and then place the seeds as they are to test the germination rate and germination power. Example 2): For example, even non-diapause seeds are seeds of the type that do not project radicles in about 1 week under a normal water-fed atmosphere, and large seeds include seed coat removal and seed slices. It is advisable to test the germination rate and germination force by carrying out the procedure. Example 3): For example, in the case of a type of seed that is difficult to remove the seed coat and cut off the seed with small seeds, phytohormones (GA,
BA, GA + BA, ethylene gas, or mixed gas) is added, and in the case of seeds of a type that still cannot attract germination within about one week, it is placed in combination with potassium nitrate solution and the germination rate and germination ability are tested. Good. In addition, oxygen substitution may be used together if necessary.

(4-3-2) Example of application to primary dormant seed group Example 1): For example, primary dormant awakening of broad-leaved tree seeds generally takes two to three months in the natural environment under humid low temperature conditions. And proceed. Therefore, some processing is indispensable for the germination test of seeds in this group, but in the case of large cleavable seeds, a seed slice containing an embryo of some form is used, and plant hormones (GA, BA, GA + BA , Ethylene gas, or mixed gas) is added, and oxygen substitution may be used together if necessary.

Example 2): For example, in a seed that can be cut in a medium size, but cannot remove the combined portion of “radicle + hypocotyl + cotyledon” from the structure of the seed, only the outer seed coat is removed,
Add plant hormones (either GA, BA, GA + BA, ethylene gas, or mixed gas) or place them with an inexpensive dormant breaker thiourea solution to determine the degree of increase in fresh weight and the ratio of radicle protrusion and cotyledon expansion. It is recommended to test the germination rate and germination power. In addition, oxygen substitution may be used together if necessary.

Example 3): For example, in the case of small seeds which cannot be cut into seed pieces or exfoliated the seed coat, it is basically necessary to add a plant hormone or, if necessary, to use oxygen substitution in combination. In addition to adjusting the concentration of plant hormones to an optimum level so that some germination phenomena can be observed before and after, combining plant hormones with potassium nitrate solution as necessary, enhance oxygen absorption efficiency with them,
In order to reduce the pressure exerted by the seed coat on the growth of embryos, the seed coat may be put in a rice polishing machine and agitated so that the seed coat surface is scratched only.

(4-3-3) Example of application to primary dormant seed group containing only buds Example 1): For example, only buds are dormant, but as soon as they fall to the ground, the radicles are extended to absorb water. However, even if the seed has the property of overwintering, in the case of a seed with a slow radicle protrusion rate, a seed slice / block containing a radicle part is prepared by cutting the seed or further cutting the side of the seed slice. It is recommended to test germination force from the rate of radicle elongation under water-supplied atmosphere. If it is necessary to actually observe the germination of the young buds, it is advisable to add the above-mentioned GA or further replace oxygen in the atmosphere.

Example 2): For example, when the radicles of seeds have already been elongated or have died after being elongated, not only seed seeds are blocked but also radicles of Removed from the roots, non-elongating individuals remove the tip of radicle tissue and place with auxin solution,
It is advisable to test the germination rate and germination power from the number of adventitious roots that differentiate from the cutting surface and their total length.

(4-4) Example of Application to Hard Seed Seed The hard seed type seed has the embryo completely developed and post-ripening is also completed.
It is typical that the seed coat is not able to germinate due to its firmness, but in any of the above tree seeds, the hardness and thickness of the seed coat causes the seed coat to exert a certain mechanical pressure on the germination and at the same time, there is a difference in degree. In fact, the fact is that it interferes with the absorption of water and oxygen. In some cases, a germination inhibitor may be included to prevent the progress of germination, but here, the adaptation to typical hard seed type seeds is limited to the following (4-4-1) to (4-4 -3) Arrange as follows.

(4-4-1) For example, seed coat strength variations are usually present in the seed population, and in order to avoid such variations, seed sections containing embryos or embryo radicle tissues are prepared. Then
It is recommended to test the germination ability from the degree of germination (radicle elongation rate and pigment formation in cotyledon tissue) in a water-supplied atmosphere.

(4-4-2) For example, in the case of seeds of the type in which the germination phenomenon cannot be detected after about one week by the method of (4-4-1), the above-mentioned seed slices are used to remove plant hormones (GA, BA,
It is advisable to test the germination rate and germination power by observing the degree of germination by adding GA + BA) or placing it under high oxygen partial pressure.

(4-4-3) For example, because the seeds are small,
If removal of outer and inner seed coats or cutting of seeds is difficult,
Dilute the seed or inner seed coat by immersing the seed in concentrated sulfuric acid or the like in the case of strong acid, or sodium hypochlorite in the case of strong alkali, or by physically damaging a part of the outer or inner seed coat. You should put it on the floor.

[0134]

The effects of the present invention are, firstly, to realize an early test method of seed germination rate applicable to all kinds of woody plant seeds (especially broad-leaved tree seeds). Secondly, based on this, it becomes possible to make a quality proof method of the used seed population or the traded seed population. That is, the germination rate of the seed population can be estimated directly or indirectly by this assay method, and seed quality assurance and handling guidelines can be provided. Thirdly, based on the estimated seed germination rate, it is possible to present the most rational construction plan that responds to the construction orderer by setting the appropriate seeding and mixing amount of woody seeds in the greening work by the sowing method. become. In addition, since it is the recognition of seed quality after being familiar with the various structural characteristics unique to each woody plant seed and the physiological characteristics regarding diapause / deterioration, the quality assurance obtained here is used as the basis. The design / construction plan to be disclosed will have higher certainty.

As a concrete effect, according to the present invention, since the proportion of individuals having the germination ability of the target plant can be measured within a short period of 7 days, individuals having an accurate germination ability without causing seed deterioration. It is possible to efficiently and quickly measure the ratio. In addition, depending on the characteristics of the seed, it is possible to cut the seed tissue with the outer seed coat and inner seed coat attached, or to place the seed seed with the outer seed coat and inner seed coat attached, so the majority of the seeds can be removed by conventional extraction. Unlike the embryo method, it is not necessary to completely expose the embryo to all kinds of seeds, and workability can be greatly facilitated.
Further, the removal of outer seed coat and inner seed coat, there is also a type of seeds that require immersion treatment before cutting the seeds, according to the present invention, the immersion treatment period for any type of seed is overnight. Because it is good, the test period will be greatly shortened.

In the greening work, because the nature of handling living things called seeds limits the construction period, it is required to measure the germination rate promptly before construction. This method is an extremely effective method in such a case, and as a practical problem, it can greatly contribute to society as an alternative to the conventional germination test method, which has been difficult to apply to tree plant seeds. In particular, the method of the present invention can assay the germination rate within 7 days at the longest in all cases. Therefore, it became possible to test the germination rate within a short period of 7 days even for seeds having strong dormancy, which had been difficult to apply the germination test. This is an extremely effective method when it is necessary to confirm the quality of seeds in advance as soon as possible, such as greening work that uses seeds naturally collected by local residents. In addition to seeds used for greening work, in the fields of seedling production, etc., efficient seedling production can be performed by using this method to test the germination rate.

By certifying the quality of the seed using this germination test, it becomes possible to certify the quality of the seed with higher accuracy and earlier than ever before. In addition, by setting the seeding rate of woody plant seeds in the greening work using this test result (quality proof), it is easy to incorporate local tree seeds into the design, and as a result, it is possible to ensure that seeds are not wasted. High greening work can be realized.

[Brief description of drawings]

FIG. 1 is a flow chart schematically showing the procedure of an early assay method for woody plant seeds according to the present invention.

FIG. 2 is a flow chart schematically showing a procedure of a method for early certification of woody plant seeds according to the present invention.

FIG. 3 is a graph showing an example of a calibration curve showing a correlation between a germination force value and a germination rate created by a calibration curve creating step of the method for certifying early quality of woody plant seeds according to the present invention.

FIG. 4 is a flow chart schematically showing a procedure of a method for designing seeding amount of woody plant seeds according to the present invention.

Front page continuation (72) Inventor Toshiyuki Akimoto 10 Goi Kaigan, Ichihara-shi, Chiba Japan Joint Fertilizer Co., Ltd., Chiba factory (72) Inventor Yohji Esashi 10-24 Otemachi, Aoba-ku, Sendai-shi, Miyagi (56) Reference Reference BABELEY GS, Journal of Japan Forest Society, Japan, 1986, Vol. 68, No. 5,197-199 (58) Fields investigated (Int.Cl. ⁷ , DB name) A01C 1/00-1/08 A01G 7/00 JISST file (JOIS)

Claims (18)

(57) [Claims] 1. In an early method for assaying woody plant seeds, the seeds of the woody plant seeds are reduced depending on the structural characteristics of the woody plant seeds.
At least completely immature embryo seed, immature embryo seed, embryo mature seed
Offspring, classified into any of the solid seed type, the pretreatment method of the seed, the bed placement method and the treatment method are appropriately selected according to the classification of the seed and the physiological characteristics of the seed, and the selected pretreatment Process the seeds by the method and the bed placement method, the processed seeds are placed on the germination test bed under continuous light irradiation conditions and constant temperature conditions by the selected treatment method, and the seeds that have been placed are germinated. An early method for assaying seeds of woody plants, which comprises measuring the proportion of individuals having ability and / or germination ability. 2. A premature assay woody plant seeds, the seeds in accordance with the structural properties of the woody plant seed, small
At least completely immature embryo seed, immature embryo seed, embryo mature seed
Offspring, classified into any of the solid seed type, the pretreatment method of the seed, the bed placement method and the treatment method are appropriately selected according to the classification of the seed and the physiological characteristics of the seed, and the selected pretreatment Process the seeds by the method and the bed placement method, the processed seeds are placed on the germination test bed under continuous light irradiation conditions and constant temperature conditions by the selected treatment method, and the seeds that have been placed are germinated. The proportion and / or germination capacity of capacity-bearing individuals is measured, and the correlation between the measured proportion or germination capacity of germ-bearing capacity individuals and the actual germination rate of the seeds is derived in advance .
Can, early assays woody plant seed, characterized in that to estimate the actual germination rate of the seed from the ratio or germination of the germination held individual by applying the correlation. 3. The method of placing a seed classified as the immature seed of the complete embryo is selected from at least one of the following methods (1-A) to (1-F): Claim 1
Or an early method for assaying seeds of woody plants according to item 2 . (1-A): A method in which the endosperm tissue is cut so as to contain the embryo and the seed slice is placed after removing the inner seed coat. (1-B): A method in which a seed slice obtained by cutting the endosperm tissue so as to contain the embryo is placed with only the inner seed coat attached. (1-C): A method in which a seed slice obtained by cutting the endosperm tissue so as to contain the embryo is placed with the outer seed coat attached. (1-D): After cutting the endosperm tissue by any of the methods (1-A) to (1-C) above, a seed section obtained by cutting the side surface of the endosperm tissue so as to further contain an embryo is placed. Method. (1-E): A method of removing the embryo from the seed and placing it on the bed. (1-F): When it is difficult to remove the outer seed coat or inner seed coat and cut the seed, after soaking the seed in strong acid or strong alkali,
Alternatively, a method in which the outer seed coat or the inner seed coat is physically damaged and subsequently placed on the floor. 4. The method of placing a seed classified as the immature embryo seed is selected from at least any of the following methods (2-A) to (2-I). Item 1 or
2. An early test method for woody plant seeds described in 2 . (2-A): A method in which the endosperm tissue or the endosperm tissue and the cotyledon tissue are cut so as to include the embryo or the radicle root tissue of the embryo after removing the inner seed coat, and a seed section is placed. (2-B): A method in which the endosperm tissue or the endosperm tissue and the cotyledon tissue are cut so as to include the embryo or the radicle root tissue of the embryo with only the inner seed coat attached thereto. (2-C): A method in which an endosperm tissue or endosperm tissue and cotyledon tissue are cut so as to contain an embryo or an embryo radicle root tissue with an outer seed coat attached thereto. (2-D): after cutting the endosperm tissue or endosperm tissue and cotyledon tissue by any of the methods described in (2-A) to (2-C) above, the embryo or radicle root tissue of the embryo should be further included. A method of implanting a seed section obtained by cutting the side surface of the endosperm tissue or the endosperm tissue and the cotyledon tissue. (2-E): A method of removing the embryo from the seed and placing it on the bed. (2-F): A method of removing the inner seed coat and then placing the seeds without cutting them. (2-G): A method in which seeds are placed as they are without cutting, with only the inner seed coat attached. (2-H): A method in which seeds are placed as they are without cutting the seeds with the outer seed coat attached. (2-I): When it is difficult to remove the outer seed coat or inner seed coat and cut the seed, after soaking the seed in a strong acid or a strong alkali,
Alternatively, a method in which the outer seed coat or the inner seed coat is physically damaged and subsequently placed on the floor. 5. The method of placing a seed classified as the embryo mature type seed is selected from at least one of the following methods (3-A) to (3-I). Item 1 or
2. An early test method for woody plant seeds described in 2 . (3-A): A method in which seed seeds obtained by cutting the cotyledon tissue so as to contain the radicle tissue of the embryo are placed after removing the inner seed coat. (3-B): A method in which a seed section obtained by cutting the cotyledon tissue so as to contain the radicle tissue of the embryo is placed with only the inner seed coat attached. (3-C): A method in which a seed section obtained by cutting the cotyledon tissue so as to contain the embryo radicle tissue is placed with the outer seed coat attached. (3-D): Seed section obtained by cutting the cotyledon tissue by any of the methods (3-A) to (3-C) above, and further cutting the side surface of the cotyledon tissue so as to include the radicle tissue of the embryo How to place the floor. (3-E): A method of removing the inner seed coat and then directly placing the seed without cutting. (3-F): A method in which seeds are placed as they are without cutting, with only the inner seed coat attached. (3-G): A method in which seeds are placed as they are without cutting the seeds with the outer seed coat attached. (3-H): In the case of an individual rooted immediately after seeds set and dropped or an individual rooted easily during storage, an already rooted root does not root from a root. Individuals are excised from the tip of radicle tissue and then placed on a germination bed containing auxin. (3-I): When it is difficult to remove the outer seed coat or inner seed coat and cut the seed, after immersing the seed in a strong acid or a strong alkali,
Alternatively, a method in which the outer seed coat or the inner seed coat is physically damaged and subsequently placed on the floor. 6. The method of placing seeds classified as the hard seed type seeds is selected from at least one of the following methods (4-A) to (4-D). Item 1 or 2
An early test method for woody plant seeds as described in 1. (4-A): The seed according to claim 3, which is a solid seed but is not difficult to remove the outer seed coat or inner seed coat or cut off the seed, and can be structurally classified as an immature seed of complete embryo. A method of implanting the immature embryo of a complete embryo by any one of (1-A) to (1-E). (4-B): a KataMinoru type seed but rather difficult to cut the removal or seed of the outer seed coat or inner seed coat, if structurally classified into embryonic immature seeds, the embryo of claim 4 A method of placing an immature seed by any one of the methods (2-A) to (2-G). (4-C): The embryo maturation according to claim 5, when the seed is a hard seed type but removal of the outer seed coat or inner seed coat or cutting of the seed is not difficult and the seed can be structurally classified as an embryo mature seed. A method of placing the seeds by any of the methods (3-A) to (3-F) in the type seeds. (4-D): When it is difficult to remove the outer seed coat or inner seed coat and cut the seed, after soaking the seed in any one of strong acid, strong alkali, and boiling water, or physically removing the outer seed coat or inner seed coat How to put the floor after scratching. For 7. processed seeds, early assays woody plant seed according to any one of claims 1 to 6, characterized in that the processed seeds to water wash prior to plated . 8. The method for treating the seed is physiological
Dormant seeds, seeds with slow embryo growth, or ectodermal
And seeds for which it is difficult to remove inner seed coat or cut seeds.
And, after adjusting the solution or mixed solution of cytokinin or gibberellin concentration and mixing ratio suitable for the seeds, which of claims 1-7, characterized in that the plated from in addition to the germination test bed An early method for assaying seeds of woody plants according to any one. 9. The method of treating the seed includes embryo maturation
Mold seeds or solid seeds that share this mold and are physiological
Removal of dormant seeds or outer and inner seed coats
For seeds that are difficult to cut , adjust the concentration and mixing ratio of a solution of thiourea or potassium nitrate or a mixed solution thereof suitable for the seed, and then add this to the germination test bed and then place the seed. Claims 1 to 7 characterized
An early test method for woody plant seeds according to any one of 1. 10. The method for treating the seed comprises physiological treatment
Dormant seeds, seeds that need to promote embryonic growth
Removal of offspring, or outer and inner seed coats or cutting of seeds
To hard seed, early assays woody plant seed according to any one of claims 1 to 7, characterized in that placing the germination test bed with high oxygen partial pressure. 11. The method for treating the seed comprises physiological treatment.
Dormant seeds, seeds that need to promote embryonic growth
Removal of offspring, or outer and inner seed coats or cutting of seeds
For difficult seeds, the germination test bed is placed under ethylene gas, a mixed gas of carbon dioxide added to ethylene gas, or a mixed gas of carbon dioxide and oxygen added to ethylene gas. 8. An early assay method for woody plant seeds according to any of 7 . 12. A fungus as a pretreatment method for the seed
The method for early detection of seeds of woody plants according to any one of claims 1 to 12 , characterized in that the seeds that are easily invaded or adhered are sterilized. 13. A method of pretreating the seeds, which comprises absorbing water.
For seeds that facilitate seed coat removal or seed cutting by
And, early assays woody plant seed according to any one of claims 1 to 11, characterized in that treating immersion water overnight the seeds. 14. A surface as a pretreatment method for the seed
The oil and fat component on the surface of the outer seed coat or the inner seed coat of the seed is removed from the seeds to which the oil and fat component is attached to the seed of the woody plant seed according to any one of claims 1 to 11. Assay method. 15. The measuring method according to any one of a) to d) below is selected when measuring the proportion of individuals having germination ability with respect to the seeds placed on the beds.
14. An early assay method for woody plant seeds according to any of 14 . a): A method of measuring the proportion of individuals with elongated radicle tissue or individuals with radicle protruding from seed tissue. b): A method of measuring the proportion of individuals whose radicle tissue is curved in the direction of gravity. c): A method of measuring the proportion of individuals in which the cotyledonary tissue is swelled while being firm, or the proportion of individuals in which the cotyledonary tissue has chlorophyll formed and turned green. d): A method of measuring the proportion of individuals with adventitious roots formed. 16. The following i) to v) when measuring the germination ability of the individual having the germination ability with respect to the seeds placed on the floor:
Early assay woody plant seed according to claim 1-14 in which any of the measurement methods are being selected. i): A measuring method of measuring the length of radicles that have grown within a certain period or converting the length of the radicles per day. ii): A measuring method of measuring the weight of the embryo increased within a certain period or converting the weight of the embryo per day. iii): A method for measuring the proportion of individuals with germination ability measured by the method according to any one of claims 16 to 16 within a certain period, which is expressed as a percentage. iv): A measuring method of measuring the length of adventitious roots that have grown within a certain period or converting the length of adventitious roots per day. v): A measuring method for measuring the degree of chlorophyll formation within a certain period or converting the degree of chlorophyll formation per day. 17. An early quality certifying method for woody plant seeds, which comprises certifying the quality of seeds by presenting at least one of the following a) or a). A): Proportion of individuals having germination ability and / or germination ability obtained by the early assay method of the woody plant seed according to any one of claims 1 to 16 . B): The actual germination rate estimated from the proportion of individuals having germination ability or germination ability obtained by the early assay method for the woody plant seed according to any one of claims 1 to 16 . 18. A method for designing the seeding amount of woody plant seeds, which comprises calculating a unit seeding amount of woody plant seeds in a sowing process by using at least one of the following a) to c). A): Proportion of individuals having germination ability obtained by the early assay method for woody plant seeds according to any one of claims 1 to 16 . B): The actual germination rate estimated from the proportion of individuals having germination ability or germination ability obtained by the early assay method for the woody plant seed according to any one of claims 1 to 16 . C): The germination rate presented by the quality certification method according to claim 17 .