KR100978888B1 - Germinating Method for Camptotheca acuminate - Google Patents

Abstract

The present invention relates to a method of germinating a rare tree seed, and more specifically, to sprout a rare tree seed through one or more treatments selected from the group consisting of treatment of a growth hormone at an appropriate concentration, maintaining an appropriate temperature during germination, and controlling appropriate storage conditions. The present invention relates to a seedling germination method, wherein the seedling germination is significantly increased.

Description

Germinating Method for Camptotheca acuminate}

The present invention relates to a method of germinating a rare tree seed, and more specifically, to sprout a rare tree seed through one or more treatments selected from the group consisting of treatment of a growth hormone at an appropriate concentration, maintaining an appropriate temperature during germination, and controlling appropriate storage conditions. The present invention relates to a seedling germination method, wherein the seedling germination is significantly increased.

A rare tree is a dicotyledonous plant, a subtropical deciduous broad-leaved tree belonging to the genus Nyssaceae and native to China. Branches of rare trees do not exist in the lower part, but usually hang at the apex. They bloom in June and bear yellow-brown fruits in November. Since ancient times, eating fruits and leaves can be used to cure bad diseases and recover quickly, and eating fruits to remove toxic and anti-cancer effects. In Korea, it is called pleasing tree tree (樹). It has been used for a purpose. In recent years, it has been found that a large amount of camptothecin, an anticancer agent, is contained in leaves, stems, branches, bark, and the like, and its pharmacological utility is increasing.

However, the rare wood is originated in China, and in Korea, where soil and climate are different, the germination rate is low and the pharmacological use of the rare wood is restricted.

In order to solve the low germination rate problem of the rare trees as described above, the present inventors have completed the present invention by developing a germination method that can significantly increase the germination rate of rare trees as a result of repeated studies under various treatments and conditions.

Accordingly, an object of the present invention is to provide a rare tree seed germination method characterized by remarkably enhancing the germination rate by treating the rare tree seed with 0.1 to 10% by weight of plant growth hormone.

Still another object of the present invention is to provide a rare tree seed germination method, which is characterized in that the germination rate is significantly enhanced by sowing and germinating the seedling tree seed by adjusting the seeding temperature to 20 ° C. to 28 ° C.

Still another object of the present invention is to provide a rare tree seed germination method characterized by remarkably enhancing the germination rate by storing the rare tree seeds at -7 ° C to 10 ° C and then seeding and germinating them.

Rare trees are mainly grown in temperate climates, which are sown in Korea in November to December, and are usually sown in March of the following year. As mentioned above, rare trees are very useful in the pharmaceutical field because they are an important source of camptothecin, which is very useful as an anticancer agent. However, rare trees are plants that grow in subtropical climate. In Korea, which has a temperate climate, the germination rate is very low because not only the climate is not right, but also the fall comes early and there is not enough fruit of seeds. This low germination rate is a limiting factor in the production of pharmacologically active substances such as camptothecin from the rare trees.

Thus, the present inventors conducted a study to find out the treatment conditions for increasing the germination rate of the rare tree, and completed the present invention by finding the germination conditions of the rare tree seed germination can obtain at least two times the germination rate than the existing germination rate.

The present invention relates to a rare tree seed germination method characterized by significantly increasing the germination rate of rare tree seeds, including the step of germinating by treating the plant growth hormone to an appropriate concentration. In this regard, the present inventors treated seedlings with concentrations of gibberellin (GA ₃ ) and auxin, known as plant growth hormone, and naphthaleneacetic acid (NAA), which increased the concentration of both rare hormones, compared to no treatment. Seed germination rate was shown, especially in case of GA ₃ it was found that the seed germination rate is significantly superior to NAA.

Therefore, the plant growth hormone that can be used for the seedling germination method according to the present invention is a gibberellin hormone (eg, GA ₁ , GA ₂ , GA ₃ , GA ₄ , GA ₅ , GA ₇ , GA ₈ , GA ₁₂ , etc.) And auxin hormones (eg, indole acetic acid (IAA), indole butyric acid (IBA), naphthalene acetic acid (NAA), etc.), may be one or more selected from the group consisting of, preferably a gibberellin-based hormone, more preferably May be GA ₃ . In addition, the preferred plant growth hormone treatment concentration is 0.1 to 10% by weight, more preferably 0.5 to 7% by weight, more preferably 1 to 5% by weight, still more preferably 3 to 5% by weight. The concentration range of the plant growth hormone as described above is obtained experimentally, it is very important to treat in the concentration range because its action effect is very sensitive depending on the concentration. In addition, the treatment time of the plant growth hormone and the above can be appropriately adjusted according to the treatment concentration, for example, the higher the treatment concentration in the range of 30 seconds to 30 minutes, the shorter the treatment time, the lower the treatment concentration, the longer the treatment time can do. Treatment of the plant growth hormone as described above may be by any method commonly used, for example, can be carried out by a method such as dipping, spraying, coating, sealing, volatilization.

In another aspect, the present invention provides a method for germinating rare tree seed, characterized in that by increasing the seed seeding temperature to increase the germination rate of the rare tree seed. The present inventors measured the germination rate by low temperature seeding (18 ° C.), medium temperature seeding (22 ° C.) and high temperature seeding (26 ° C.) under different seeding conditions. Increasing findings revealed the optimum seeding temperatures for high germination rates.

The seeding temperature in the rare-tree seed germination method which concerns on this invention is 20-28 degreeC, It is good that it is 22-26 degreeC more preferably. When the seeding temperature is within the above range, after sufficient swelling of the seeds occurs, the seeds may germinate to obtain a high germination rate.

In another aspect, the present invention provides a method for germinating rare tree seed, characterized in that to increase the germination rate of the rare tree seed by controlling the storage temperature of the rare tree seed. The inventors have found that germination rates are higher when stored rare seedlings at lower temperatures than when stored at room temperature. The storage temperature of the rare tree seed according to the present invention is suitable for the soil temperature below the ground from November to April of Korea, and may be about -7 ℃ to 10 ℃, more preferably about 0 to 4 ℃. . If the storage temperature is lower than the above range, the seed may freeze and the germination rate may be lowered. If the storage temperature is higher than the above range, the germination may occur in an incomplete environment during the storage period, so the storage temperature of the seed may be the same as the above range. .

In a more preferred embodiment, the seed storage environment should be similar to the soil environment of the open field in order to maintain and enhance the vitality of the seed by providing an environment in which the seed breathes sufficiently and in contact with adequate moisture. For example, in order to provide adequate breathing of the seed and to provide adequate moisture, it is recommended to store it with wet sand with a water content of 10 to 30% by weight, and the wet sand where the rare tree seeds are stored is about the weight of the rare tree seed weight. It is preferably used in an amount of 2 to 5 times, preferably 2.5 to 4 times.

In addition, the storage period is from the harvesting time of the rare tree seed to the sowing time, and usually the harvesting time is from November to December, and the sowing time is about March to April the next year. Although it may be appropriately adjusted according to the specific seeding time and sowing time, preferably not more than 180 days, 60 to 180 days, preferably 90 to 150 days. The rare tree seed may be stored immediately after harvesting, but it is more advantageous to improve shelf life after being soaked and dried for a suitable period at a suitable temperature after harvesting. It is preferable that the shade temperature is 5 to 15 ° C, and the shade period is 3 to 10 days. In addition, during storage, the temperature conditions should be kept as low as possible, it is better not to leave more than 48 hours continuously below -4 ℃.

Such temperature conditions and the water content in the sand are artificially adjusted and maintained, or 60 to 180 days during sowing after seeding, where the rare tree seeds and sand are mixed at the above mixing ratio, that is, from November to April. It may also be naturally met by buried and stored in open soil soil about 5-20 cm deep from the ground for a period of time. In particular, in the case of storage by the open-air store, it provides the highest germination rate because the natural supply of sufficient air and adequate moisture, and other environmental conditions can be naturally matched.

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Seed germination method according to the present invention can further improve the seed germination rate by seeding the seedling tree after removing the outer part of the seedling except for the pear-shaped belly portion. The rare tree seed is a slender oval and the abdomen is located in the middle line of the long diameter of the ellipse. The rare tree seed used in the present invention may be one having a length of about 1 to 3 mm including the middle line in the long radial direction in which the ship is located, and the remaining part being removed in a direction parallel to the line. When the seed portion is removed as described above, the lead germination of the skin of the bark is removed, so that the supply of moisture and oxygen is smooth, so that the seed germination rate may be increased.

In another aspect, the present invention provides a rare tree cultivation method comprising the seed germination step as described above. In the above cultivation method, the step except for the seed germination step is by a method commonly known in the art.

Although rare trees produce pharmaceutical useful substances such as camptothecin and are very important pharmaceuticals, the seed germination rate is very low in Korea, but it has been a major limitation in the production of camptothecin using the same, but the rare tree seed according to the present invention. This germination method can solve this problem by remarkably increasing the germination rate of rare tree seeds.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

[ Example ]

Example  One: Rare tree  Germination rate of seeds

Seed germination rates were compared between seeding (straight) and seeding in spring (2006.3.20, or April 10, 2006) in the following year (spring onion) in order to find the appropriate seeding conditions for rare tree seeds.

Seeds were harvested on December 10, 2005 from three weeks of rare trees of about 30 years of age, native to China, in Cheolipo Arboretum for use in this example. The seeds used in the following spring were sown on the sowing date after air storage at room temperature (5-15 ° C.). As a seeding container, a ground skin port was used, and the number of seeding grains per port was 2 to 3 grains. The survey period was from December 10, 2005 to September 30, 2006.

To check the germination rate of seeds, seeding trays containing 30 pieces per block are repeated three to three times per pot from immediately after harvesting (Dec. 10, 2005) to the following spring (2006.3.20, or April 10, 2006). The germination rate of each seeding day was investigated by sowing 90 ports per treatment.

Test scale: 3 treatments × 3 repetitions × 30 per iteration = 270 pots

The germination rate showing the number of seeding and germination in the experiment and the ratio (germination rate, unit:%) of germination to the number of seeding are shown in Table 1 and FIG. 1 below.

TABLE 1

sowing
Time Repeat group I Repeat group II Repeat group III Total germination rate sowing germination Germination rate sowing germination Germination rate sowing germination Germination rate sowing germination Germination rate 12.10 30 11 37 30 10 33 30 9 30 90 30 33 3.20 30 7 23 30 8 27 30 9 30 90 24 27 4.10 30 8 27 30 7 23 30 8 30 90 23 26 Average 30 8.7 29 30 8.3 27.6 30 8.7 30 90 25.7 28.6

As can be seen from the above results, the germination rate was higher than that of the spring sowing in the spring of the following year, but since it has to grow through winter after germination, the growth conditions are not appropriate and do not grow normally. For this reason, direct cultivation is not appropriate due to the climate of Korea. In the case of spring, the low germination rate is presumed to be due to loss of nutrients in the body due to seed respiration during storage period of the seed. Therefore, we reaffirmed the need to develop new seed germination methods to reduce or compensate for these losses during the seed period.

Example  2: Seed germination according to plant growth hormone treatment

2006.12.10 Gibberellin GA3 and NAA were treated for 3% and 5% of each treated seedlings, and 100 grains (20 grains / sowing dish, 5 sowing plates) for each treatment. The germination rate was examined for 50 days from 2007.3.10 to April 30, 2007. The gibberellin GA3 and NAA treatment was performed by dissolving gibberellin GA ₃ and NA in sterile water to the concentration, soaked after 5-30 minutes so that the seeds are sufficiently immersed, washed with sterile water and put the seeds in airy well.

A seeding dish (Pat Dishes) was used as a seeding container, and a cotton ball soaked in distilled water was placed thereon and irradiated with water every day. The investigation period was 50 days from 2007.3.10 to April 30, 2007, and the test scale was 100 tablets / treatment × 2 hormone species treatment × 3 hormone concentration treatment × 3 repetition = 1,800 tablets.

The germination rate (%) expressed as the seeding number, germination number and the ratio of germination number to the seeding number in the experiment is shown in Table 2 below.

TABLE 2

hormone Repeat group I Repeat group II Repeat group III Total germination rate Kinds density sowing germination Germination rate (%) sowing germination Germination rate (%) sowing germination Germination rate (%) sowing germination Germination rate (%)
GA ₃ 0% 100 6 6 100 5 5 100 7 7 300 18 6 3% 100 45 45 100 44 44 100 42 42 300 131 43 5% 100 44 44 100 43 43 100 39 39 300 112 37
NAA 0% 100 6 6 100 7 7 100 6 6 300 19 6 3% 100 12 12 100 15 15 100 13 13 300 40 13 5% 100 10 10 100 11 11 100 12 12 300 33 11

As shown in Table 2, the germination rate was increased when the plant growth hormone treatment than the non-treatment, in particular, the GA3 treatment significantly improved the degree of germination.

Example  3: germination rate by sowing temperature

2006.12.10 After storing cultivated rare tree seeds in the open-air store, repeat them three times (20 grains / sowing plate, 5 sowing plates) at each treatment temperature (18 ℃, 22 ℃, and 26 ℃) by each sowing temperature. The germination rate was examined for 50 days from 2007.3.10 to April 30, 2007.

A seeding dish (Pat Dishes) was used as a seeding container, and a cotton ball soaked in distilled water was placed thereon and irradiated with water every day. The investigation period was 50 days from 2007.3.10 to April 30, 2007. The test scale was 100 grains / treatment × 3 temperature treatment × 3 repetitions = 900 grains.

The germination rate (%) expressed as the seeding number, germination number and the ratio of germination number to the seeding number in the experiment is shown in Table 3 below.

 [Table 3]

sowing
Temperature Repeat group I Repeat group II Repeat group III Total germination rate sowing germination Germination rate (%) sowing germination Germination rate (%) sowing germination Germination rate sowing germination Germination rate (%) 18 ℃
(Low temperature sowing) 100 5 5 100 3 3 100 4 4 300 12 4 22 ℃
(Medium temperature sowing) 100 35 35 100 38 38 100 42 42 300 115 38 26 ℃
(High temperature sowing) 100 36 36 100 37 37 100 39 39 300 112 37

As shown in Table 3, the germination rate was significantly increased in the case of medium or high temperature sowing than in the case of low temperature sowing. At low temperature of 18 ℃ or less, the germination rate was 4% and the water swelling was sufficient, but most germination did not occur. Germination began after swelling of water for about 1 month at the sowing temperature of 22 ℃ and 26 ℃ and germination rate was increased to 38% and 37%. Therefore, it was proved that the seeding temperature of a rare tree seed should be 20 degreeC or more, More preferably, it is good to set it to 22 degreeC or more.

Example  4: germination rate by storage condition

2005.12.10 The seeded rare tree seeds were dried in the shade for 5 days under the temperature condition of 5-15 ℃, stored in the following three ways, and then sown in 2006.3.20 (Storage period: December 10, 2005 ~ 2006.3) .20), germination rates 60 days after planting date were examined.

1) Air storage: Put the rare tree seeds dried in the shade in a plastic bag and store at room temperature (5 to 15 ℃),

2) Low temperature storage: Mixed wet sand and rare tree seeds with a moisture content of 70% in a weight ratio of 3: 1 and stored in a low temperature refrigerator at 0-4 ° C.,

3) open-air storage: store sand and seeds 3: 1 in the open-air (use the inclined soil net, easy to drain and manage, put the onion net again and dig a pit next to the greenhouse in a plastic planter to prevent damage to rodents) Covered and buried with some convex soil.)

As a seeding container, a grounding paper pot was used, and the number of seeding grains was 2 to 3 grains per pot. The test scale was 3 storage treatments x 3 repetitions x 30 per repetition = 270 ports.

 The germination rate (%) expressed as the seeding number, germination number and the ratio of germination number to the seeding number in the experiment is shown in Table 4 and FIG. 2 below.

 [Table 4]

How to save Repeat group I Repeat group II Repeat group III Total germination rate sowing germination Germination rate sowing germination Germination rate sowing germination Germination rate sowing germination Germination rate Storage 30 9 30 30 8 27 30 6 20 90 23 26 Open air store 30 16 53 30 13 43 30 12 40 90 41 46 Cold storage 30 13 43 30 11 37 30 11 37 90 35 39

As shown in Table 4 and Figure 2, the germination rate of the rare tree seeds by storage method showed an average germination rate of 37.0%, 26% in air storage, 46% in open-air storage treatment, 39% in cold storage, Compared to the low temperature storage and the open-air store, the germination rate was improved. The reason for this is that the wet sand mixed at 3: 1 maintains sufficient moisture and provides oxygen for respiration, thus facilitating the stratification in favor of germination by maintaining seed vitality during storage.

1 is a graph showing the germination rate of each seeding period of the rare tree seeds.

Figure 2 is a graph showing the germination rate by storage method of rare tree seeds.

Claims (11)

delete delete delete delete delete delete In the germination method of a rare tree seed, A rare tree seed is mixed with sand having a water content of 10 to 30% by weight and 2 to 5 times the weight of the rare tree seed, and 5 to 5 on the ground of open ground soil for a period of 60 to 180 days from November to April. Store at 20 cm depth and store Sowing is carried out at a temperature of 20 to 28 ℃, Germination method of rare tree seeds. 8. The method of claim 7, wherein the storage temperature of the rare tree seed is between 0 ° C and 4 ° C. delete 8. The method of claim 7, wherein said rare tree seed is removed in a direction parallel to said line, leaving a portion of 1 to 3 mm wide, including a central longitudinal line in which the vessel is located. The method of claim 7, 8, or 10, wherein the seeding temperature of the rare tree seeds is between 22 ° C. and 26 ° C. 12.

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