KR20180042623A - Method of seminal propagation for Maesa japonica Thunb. Moritzi and Zoll - Google Patents

Abstract

The present invention relates to a mass-production method of Maesa japonica (Thunb.) Moritzi & Zoll. by establishing an optimal seed storing method and germination condition when performing seedling propagation using Maesa japonica (Thunb.) Moritzi & Zoll. seeds. In particular, it has been confirmed that germination velocity and germination rate of the Maesa japonica (Thunb.) Moritzi & Zoll. seeds can be increased by pre-processing of sealing the seeds which are dehydrated to have a moisture content of 5-10 % and by dipping and pre-treating the seeds, which are stored at a low temperature facility in a temperature condition of -20 to 4°C and relative humidity of 40%, in a mixture solution of gibberellins and potassium nitrate. Thus, the seed storage and propagation method of the present invention can be useful in preserving genetic diversity of Maesa japonica (Thunb.) Moritzi & Zoll. and securing mass-propagation.

Description

{Method of seminal propagation for Maesa japonica Thunb. Moritzi and Zoll}

The present invention relates to a method for mass propagation through seed germination of a tree, and more particularly, to a method for mass production of a tree of a tree that has established an optimum pretreatment method and a sowing temperature and a light condition before sowing of a tree.

Ville tree [Maesa japonica (Thunb.) Moritzi & Zoll.] Is an evergreen vine bush native to island size is 1 to 1.5 meters, or less shrubs, Jeju southwest altitude 120-160 m the vicinity of the closing when tree woods and forests It is distributed independently or as a cluster on the edge (Moon et al., 2006). It is classified as a rare plant with a distribution area or a small population (Moon et al., 2006).

It is necessary to develop a proper breeding method because the bamboo tree grows only in the southwestern part of Mt. Halla, and the range of the bamboo tree is narrow and the number of individuals due to the development of the area is decreased and the population is decreased due to the unauthorized harvest due to scarcity. In addition, the bille tree is composed of saponin (maesaquinone, embelin, rapanone, acetylmaesaquinone, 2-hydroxy-5-methoxy-3-pentadecenyl, tridecenyl- and tridecyl-) benzoquinone and saponins (maejaposides, maesabalides, maesasaponin and 13,28- (Ogawa and Natori, 1968; Koike et al., 1999; Foubert et al., 2009), and antioxidant and antidiabetic effects (Kim, 2014) It is necessary to establish a stable supply system by developing mass proliferation method.

However, there have been no previous studies on the cultivation and propagation of bilem trees in Korea, and there have been no studies on the propagation and cultivation of bilem trees, which are native to China in other countries, except that the seed germination proceeds slowly in the soil.

It is difficult to preserve genetic diversity because it is difficult to apply to all plants, it is costly, and only the characteristics of the sample are inherited. . Therefore, seed breeding methods are needed to economically proliferate while preserving the genetic diversity of plants.

Seed germination propagation can be applied to plants that are difficult to cut and graft, and it can preserve the genetic diversity of the plant, and at the same time it can produce large numbers of seedlings in large quantities.

However, seed germination largely depends on genetic and environmental factors. Especially, it has dormancy and it is not germinated even in suitable germination environment. Seed dormancy is caused by various causes such as imperviousness of perilla or seed coat, presence of germination inhibitor in seed coat or inhibition of growth of abdomen, and external environmental factors. Depending on the storage environment of plant species and seeds, Since the methods are different, it is necessary to develop a dormant breakout method for each plant species. In particular, it is difficult to develop a dormancy breaking method because the seeds of wild plants often enter a complex dormancy due to physiological and physical causes. If a proper dormancy breaking method is not developed, the germination rate is low, the germination rate is slow, It can not be used as an economical method for seed germination.

Gibberellin is a growth regulator that is most commonly used for seed dormancy, and it does not require a separate facility and has an advantage of improving germination efficiency through short time treatment. Gibberellin is known to be effective against the physiological dormancy of seeds, and is known to promote seed germination through the activation of hydrolytic enzymes in seeds, which promotes hydrolysis of starch to glucose. However, since gibberellin does not promote the germination of all seeds, it also inhibits germination depending on the treatment concentration and treatment method, so it is necessary to develop treatment concentration and treatment conditions suitable for plant species.

Potassium nitrate is one of the representative inorganic salts promoting seed germination and has a characteristic of facilitating moisture absorption of seeds by inducing a change in water potential. In addition, it has been reported that the oxygen requirement of the citric acid circuit is reduced in the respiration process of the seed to induce germination by promoting respiration and induce biosynthesis of auxin in the seed to promote germination. The International Seed Assay (ISTA) is usually used for seed germination by diluting to a concentration of 1 to 2 g / L, but it is known that the effects on seed usage and seed germination are different depending on plant species.

The inventors of the present invention studied germination by immersing seeds in a solution diluted with a growth regulator or inorganic salts while studying a method for mass proliferation of bilet trees. When the growth regulators and inorganic salts were treated simultaneously, the seed germination rate And the germination rate can be increased, thereby completing the present invention.

It is an object of the present invention to provide a method for mass production of a boule tree using the seeds of Maesa japonica (Thunb. Moritzi & Zoll.) To establish optimal germination rate and germination rate during breeding .

In order to achieve the above object,

1) pre-treating seeds of dried Maesa japonica (Thunb. Moritzi & Zoll.) Seeds with inorganic salts, gibberellin or a mixture thereof;

2) seeding the pretreated bilayers seed of step 1);

3) germinating the seeded villa seed of step 2); And

4) growing a seedling from the germinated seed of step 3);

To provide a viable propagation method for mass propagation of the tree.

In addition, we provide Maesa japonica (Thunb.) Moritzi & Zoll.] Plants produced by the above method.

In the present invention, seeds of Maesa japonica (Thunb.) Moritzi & Zoll.] Are immersed in an aqueous solution in which gibberellin and potassium nitrate are dissolved and then pretreated. After washing gibberellin and potassium nitrate, seeds are sown, Since the seedlings can be used for mass production of the seedlings and can be transplanted into the soil after mass propagation, the seedlings produced by the above method can produce many seedlings of uniform quality at the same time There is an advantage that it can be utilized industrially. Also, it can be used continuously as a silent breeding material through germination after seed germination in an aseptic condition, so that continuous production is possible.

Fig. 1 is a graph showing the germination rate of seeds of Bilaek seeds at different temperatures.
Fig. 2 is a graph showing the germination ratio according to the treatment of the concentration of gibberellin in the seed of Bilye seed.
Fig. 3 is a graph showing the germination ratio according to the treatment of the potassium nitrate concentration of the bille tree seed.
Fig. 4 is a graph showing a synergistic effect of germination ratio when treating gibberellin and potassium nitrate mixture of a bichon seed.
Fig. 5 is a photograph showing the growth state of Bilye seedlings by temperature.
Fig. 6 is a photograph showing the growth state according to the luminosity of the bille tree seedlings.
Fig. 7 is a photograph showing the growth state of the bille seedlings grown by the method of the present invention after 50 days from sowing.
Fig. 8 is a photograph showing a large-scale cultivation of a billy tree seedlings.

Hereinafter, the present invention will be described in detail.

The present invention

1) pre-treating seeds of dried Maesa japonica (Thunb. Moritzi & Zoll.) Seeds with inorganic salts, gibberellin or a mixture thereof;

2) seeding the pretreated bilayers seed of step 1);

3) germinating the seeded villa seed of step 2); And

4) growing the seedling from the germinated seed of step 3).

In addition, the present invention provides a tree plant produced by the method.

The drying of the step 1) is preferably carried out at a temperature of 10 to 15 ° C and a relative humidity of 5 to 10%, and it is preferable that the dried bamboo tree has a moisture content of 5 to 10% Do not.

According to the present invention, it is possible to produce viable seeds for a long time by storing low-temperature seeds of bille tree seeds for a long time, and seeds obtained after low-temperature storage or seeds immediately after harvesting can be seeded after pre-treatment. More specifically, the seeds of the bille tree were selected, and then dried in a ventilated bag at a relative humidity of 10% and a temperature of 15 ° C. until the seed moisture content was less than 5 to 10% After storing in a storage room at -18 to 4 ° C and 40% relative humidity, seeds are taken out and seeded after pre-treatment (wetting, precooling, growth regulators, inorganic salts treatment) and seeding.

The inorganic salt of step 1) is preferably potassium nitrate (KNO ₃ ), but is not limited thereto. The potassium nitrate is preferably 50 to 500 mg / L, but is not limited thereto.

If the drying temperature is lower than 10 ° C, there is a disadvantage that the seed drying speed is slow and the drying time is prolonged. Also, when the drying temperature is higher than 20 ° C, it is difficult to maintain proper moisture because the seed drying speed is fast. When the drying temperature is above 28 ° C for 30 days or more, the seed germination rate is lowered.

When the relative humidity is more than 20%, the drying speed is slowed and the efficiency is decreased. When the relative humidity is more than 60%, harmful microorganisms such as fungi and bacteria on the seeds are proliferated, . In addition, when the relative humidity is lower than 5%, the seed surface is dried quickly, resulting in deterioration of vitality or death of the seed.

When the moisture content of the seed is 10% or more, the seed may freeze and degrade or die during storage at a low temperature of 2 캜 or lower. If the moisture content of the seed is less than 5%, the seed germination is restored. Or there is a case in which the vitality of the seed is lowered or the seed is killed.

The gibberellin in step 1) is preferably in a concentration of 200 to 2000 mg / L, but is not limited thereto.

The pretreatment of step 1) may be carried out by various methods. Generally, the seeds are immersed in sterilized water, and the seeds are immersed in the sterilized water at a temperature of 2 to 6 ° C. for 12 to 48 hours, or a growth regulator such as gibberellin And a solution diluted with inorganic salts such as potassium nitrate.

It is preferable that the seeding of the step 2) is performed by seeding 15 to 20 seeds per seedling area (m ² ), and the germination and growth is preferably performed at a temperature of 23 to 35 ° C, but is not limited thereto. Germinated seedlings have the best growth at 25-35 ° C. Below 8 ° C, the leaves are yellowed or browned due to cold weather, and are exposed to low temperatures below 5 ° C.

The growth of step 4) is preferably performed under light-shielding conditions of 30 to 70% of non-sound. The germinated seedlings are excellent in growth in the half-tone with 30 to 70% non-shading condition of non-sounding. In the unshielded vinyl greenhouses or nogi, the ends of the leaves are burned, and the leaves are crumpled and smoothed.

In a specific example of the present invention, the low-temperature-stored villa seeds were dipped in sterilized water to which gibberellin and a potassium nitrate mixture had been added, and then moistened at 4 ° C for 24 hours and then germinated at 30 ° C. All germinated, and it was confirmed that the mixed treatment of dibelredin and potassium nitrate could increase seed germination rate and germination rate.

As a result of germination experiments, the best temperature for seed emergence from seeds was found, and the seeds of Bilye seed showed the highest germination rate at 30 ℃, germination at 65% at 30 days after sowing and low germination rate at low temperature (15 ℃) The final germination rate was also around 60%. On the other hand, germination was not carried out at a high temperature of 35 ° C (see FIG. 1). The inventors of the present invention found that the germination rate of the bille tree seed was higher than that of the seedlings treated with potassium nitrate and germinated with sterilized water when treated with potassium nitrate, And the germination rate was highest (see Fig. 2). As a result of treating gibberellin with the seeds of Bilye seeds, the germination rate of the seeds of Bilye seed increased with increasing gibberellin treatment concentration. Especially, when treated with 2000 mg / L, all seed germinated, (See Fig. 3). The inventors of the present invention confirmed that the mixing treatment of inorganic seeds with gibberellin and the gibberellin treatment resulted in a germination rate higher than the predicted value in all treatments of potassium nitrate and gibberellin, Reference). In particular, when 1000 mg / L of gibberellin and 200 mg / L of potassium nitrate were mixed, the synergistic effect was greatest and the germination speed was fast, which proved to be the most efficient seed germination method (see FIG. 4).

The inventors of the present invention conducted a method in which the seeds of biloba seedlings were put into a mixed solution of 200 mg / L of potassium nitrate and 10000 mg / L of gibberellin in the same manner as described above, and then washed with water after wet treatment at 4 ° C for 24 hours to remove the mixed solution, . The germination was started within 10 days of ginseng seeds germinated in a greenhouse of 23-35 ℃ at a temperature of 23-35 ℃ and non-sounding, and germinated more than 95% in 20 days and less than 4cm, 5 or more leaves, 5 cm or more of root length, and could be utilized as seedlings (see FIGS. 5 to 7). The germination seedlings pretreated by the above method had good roots growth and rooting in the soil for horticultural development, and the growth and the differentiation of the root were good, and the seedlings were grown in normal seedlings. When cultivated under the above temperature and light- After that, it was grown at a height of about 20 cm and could be sold as a landscape material. When cultivated for over one year, it could grow to over 1 m and sold as a shrub (see FIG. 8).

Therefore, the seedling of a certain stage can be rapidly produced and mass-propagated through the method of the present invention, and it can be effectively used as an effective seedling propagation method using the seed of the tree of bilemen.

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples.

It is to be understood, however, that the following examples and experimental examples are illustrative of the present invention and are not intended to limit the scope of the present invention.

Tree  Selection, drying and cold storage of fruit seeds

The fruits of Maesa japonica (Thunb.) Moritzi & Zoll.) Were collected from late September to mid October, washed with flowing water to remove the rind and pulp, and the seeds were placed in a ventilated mesh bag at 15 ° C, And dried at a moisture content of about 8% in a drying chamber having a relative humidity of 10%. The dried seeds were sealed in a glass bottle and stored in a storage room at -18 ° C and a relative humidity of 40%.

< Experimental Example  1> Tree  Seed Germination temperature  Confirm

The present inventors carried out the following experiment to determine the most suitable temperature for sprouting from seeds through germination experiments.

Specifically, the seeds of the bilayers prepared in Example 1 were taken out, 70% ethanol was added thereto, shaken for 30 seconds, firstly sterilized, then 70% ethanol was removed, 2% sodium hypochlorite solution was added and sterilized for 15 minutes . After sterilization, the sodium hypochlorite was removed by rinsing with sterilized water 5 times. The sterilized seeds were placed in sterilized water for 24 hours at 4 ° C in a humid condition and then seeded on a 1% agar medium. The seeds were sown at intervals of 0.8 × 0.8 cm in a 90 × 40 mm petri dish and germinated at 15, 20, 25, 30, and 35 ° C for 16/8 hours (light / dark)

As a result, as shown in Fig. 1, the seed germination rate was the fastest at 30 ° C, germinated 65% after 30 days of sowing, and the germination rate was extremely low thereafter. At low temperature (15 ℃), germination rate was low and the final germination rate was around 60%. On the other hand, germination did not occur at a high temperature of 35 ° C (FIG. 1). Therefore, it was confirmed that the seeds of Bilye seed were most suitable to germinate at 30 ℃.

< Experimental Example  2> Determination of germination rate by inorganic salt treatment

The present inventors conducted the following experiments to examine the germination rate of inorganic seeds treated with inorganic salts.

Specifically, the seeds of the bamboo seedlings were disinfected by the method of Experimental Example 1, and then potassium nitrate was added to the sterilized water at a dilution of 0, 50, 100, 200 and 500 mg / L, , And then germinated on a 1% agar medium at 30 ° C for 16/8 hours (day / person).

As a result, as shown in FIG. 2, the germination rate of the seedlings was found to be higher than that of the seedlings treated with potassium nitrate and germinated with sterilized water. Especially, the germination rate was highest when treated with 200 mg / L (Fig. 2).

< Experimental Example  3> Gibberellin  Identification of germination rate by treatment

The present inventors conducted the following experiments to treat gibberellin in the seeds of Bilye seeds and determine the germination rate thereof.

Specifically, after seeding the seeds of the pine trees with the method of Experimental Example 1, gibberellin (GA ₃ ) was added to the sterilized water in a diluted solution at concentrations of 0, 200, 500, 1000, 1500 and 2000 mg / And then germinated on a 1% agar medium at 30 ° C for 16/8 hours (day / month).

As a result, as shown in FIG. 3, the germination rate of the seeds of bileta seeds increased as the concentration of gibberellin treatment increased. Especially, when seeds were treated at a concentration of 2000 mg / L, all the seed germinated and the germination rate was the highest 3).

< Experimental Example  4> inorganic salts and Gibberellin  Identification of germination rate by mixing treatment

The present inventors conducted the following experiments in order to determine the germination ratio of the seeds of the bilemen seeds by mixing inorganic salts and gibberellin.

Specifically, the seeds of Bilila seed were sterilized by the method of Experimental Example 1, and then potassium nitrate (200 mg / L) and gibberellin were added to the sterilized water at the concentrations of 1000, 1500 and 2000 mg / L, respectively After wet treatment at 4 ° C for 24 hours, seeds were sown on a 1% agar medium and germinated at 30 ° C for 16/8 hours (day / person).

In order to investigate the effect of mixing of inorganic salts and gibberellin on seed germination, we estimated the germination rate using the following Colby formula and compared the actual germination rate to the synergistic effect by mixing treatment.

[Colby formula]

Germination rate predicted value (%) = (A + B) - (A * B / 100)

A: Germination rate of gibberellin single treatment

B: Germination rate of single treatment of potassium nitrate

As a result, as shown in Table 1 and Fig. 4, it was confirmed that the mixing treatment of potassium nitrate and gibberellin had a remarkably synergistic effect due to the higher germination rate than the predicted value at all treatments (Table 1). In particular, when 1000 mg / L of gibberellin and 200 mg / L of potassium nitrate were mixed, the synergistic effect was largest and the germination speed was fast, which proved to be the most efficient seed germination method (FIG. 4).

After 5 days of sowing After 8 days of sowing 10 days after sowing
single Mixed treatment single Mixed treatment single Mixed treatment Germination rate % ) Estimated value (%) Germination rate % ) Germination rate (%) Estimated value (%) Germination rate (%) Germination rate (%) Estimated value (%) Germination rate (%) GA ₃ 1000 mg / L 24 24 56 52 60 86 72 82 100 KNO ₃ 200 mg / L 0 16 34 GA ₃ 1500 mg / L 40 40 60 72 77 85 88 92 100 KNO ₃ 200 mg / L 0 16 34 GA ₃ 2000 mg / L 45 45 62 74 78 88 90 93 100 KNO ₃ 200 mg / L 0 16 34

Tree  Seed Seedlings  Growing and growing

The inventors of the present invention found that the seeds of Bilye tree seeds were put into a mixed solution of potassium nitrate 200 mg / L and gibberellin 10000 mg / L by the method of Experimental Example 4 described above and then subjected to wet treatment at 4 ° C for 24 hours, The solution was removed and then sown.

Specifically, the seeding method was carried out in the same manner as in Example 4, except that the seeds of the vilweed seedlings pretreated with the method of Experimental Example 4 were applied to horticultural soil (pH 5.5 to 7.0, EC 1.2 dS / m or less, ammonia nitrogen 400 mg / 25 to 72 plug trays or 2 to 5 pots in 5 to 8 cm pots filled with 15 to 20 spores per 0.1 m ^{2 of} seedling surface area. Respectively.

As a result, as shown in Fig. 5, when the temperature of the greenhouse was 25 DEG C or less, the germination period was long, the germination rate was low, and the germinated seedlings grew slowly. In addition, the germination rate was suppressed even at 35 ℃ or higher, the leaf size was small, the growth was delayed and the root rot was characterized. The germinated seedlings showed excellent growth at 25 to 35 ° C., and under 8 ° C., the leaves were yellowed or browned due to cold weather (FIG. 5). It was necessary to warm up in the winter, and when it was exposed to low temperature of 5 ℃ or lower for a long time, it was sacrificed.

As shown in FIG. 6, the seedlings showed excellent growth in the transplanted plants under the condition of 30 to 70% non-shading condition with no sound. In the greenhouse or the non-shaded greenhouse, the ends of the leaves were burned and the leaves were wrinkled (Fig. 6).

Therefore, the seeds of the bille tree seedlings germinated in the greenhouse of 23 to 35 ° C and 30 to 75% of the non-soundness by seeding on the horticultural soil after the pretreatment by the method of <Experimental Example 4> started germination within 10 days, %, More than 5cm in length, more than 5cm in length, more than 4cm in height, more than 5cm in length, within 50 days (Fig. 7).

The germination seedlings pretreated by the above method had good roots growth and rooting in the soil for horticulture, and the growth and leaf differentiation of the shoots were good and the seedlings grew as normal seedlings. When cultivated under the above temperature condition and light- Later, it was grown at a height of about 20 cm and could be sold as a landscape material. When cultivated for more than one year, it could grow to more than 1 m and sold as a shrub (FIG. 8).

Claims (11)

1) pre-treating seeds of dried Maesa japonica (Thunb. Moritzi & Zoll.) Seeds with inorganic salts, gibberellin or a mixture thereof;
2) seeding the pretreated bilayers seed of step 1);
3) germinating the seeded villa seed of step 2); And
4) growing a seedling from the germinated seed of step 3);
A method for propagation of the present invention for the mass propagation of the tree.
The method according to claim 1, wherein the drying of step 1) is carried out at a temperature of 10 to 15 캜 and a relative humidity of 5 to 10%.
The method according to claim 1, wherein the dried blandwood of step 1) has a moisture content of 5 to 10%.
The method according to claim 1, wherein the inorganic salts of step 1) are potassium nitrate (KNO ₃ ).
5. The method according to claim 4, wherein the potassium nitrate has a concentration of 50 to 500 mg / L.
[3] The method according to claim 1, wherein the gibberellin in step 1) has a concentration of 200 to 2000 mg / L.
The method according to claim 1, wherein the pretreatment of step 1) is carried out at a temperature of 2 to 6 캜 and under a dark condition for 12 to 48 hours.
[3] The method according to claim 1, wherein the seeding in step 2) is carried out by seeding 150 to 200 seeds per mound area (m &lt; ² &gt;).
The method according to claim 1, wherein the germination and growth are carried out at a temperature of 23 to 35 占 폚.
[3] The method according to claim 1, wherein the growth of step 4) is performed under a light-shielding condition of 30 to 70% of non-sound.
Maesa japonica (Thunb.) Moritzi & Zoll.) Plants produced by the method of claim 1.

Images