KR100444330B1 - A culture method of Centella asiatica plants - Google Patents

Abstract

The present invention provides a method of aseptically cultivating seedling plant seedlings including roots, stems and leaves in a cabin, transferring the cultured seedlings to purified soil, and purifying the seedlings. Transplanting to land with higher than 13 ℃ and average temperature higher than 13 ℃, and 6 months of cultivated cultivated on the land, the average temperature is greater than or equal to 22.3 ℃, the minimum average temperature is greater than or equal to 17.6 ℃ To provide a method of cultivating the plant stem plants comprising the step of harvesting the leaves and stems in the state, the present invention is a cultivation and cultivation of seedlings optimized for mass production of plant stem plants, shading in seedlings, transplanting between seedlings in soil transplantation By investigating the distance and timing of transplanting seedlings, it is possible to provide a way to grow cultivated plants in houses or fields. Was, very useful inventions of the pool, even if they are particularly useful bottle material revenue growth of up to farmers By harvesting conditions and the Centella asiatica plant in the optimal time to accumulate as well as to contribute to import substitution.

Description

A culture method of Centella asiatica plants

The present invention relates to a cultivation method of cultivated plants, and more particularly, to cultivate cultivated plants by cultivating them in the soil, and harvesting leaves and stems at the time when the useful substances in the cultivated plants are maximized to produce high quality therapeutic raw materials. It relates to a cultivation method of Centella plants for.

Centella asiatica (L.) Urban is a perennial crawling herb that belongs to the erectus family and grows in Southeast Asia with a subtropical climate, especially in Malaysia, India and Madekaska.

The main components of the centella are very useful pharmacological ingredients such as asiaticoside, madecassoside, asiatic acid, madecaic acid and trace alkaloids. It is excellent in the treatment of skin diseases and wounds, and is effective in anti-dementia, asthma, headache, gonorrhea, syphilis, skin disease, leprosy and gastrointestinal diseases.

However, because the grass grows mainly in high temperature and high humidity, it is extremely limited in Korea except islands of Jeju Island and southern provinces, which belong to the warm zone, and breeding is carried out through seed breeding and nutrition breeding through laparotomy. In the native area, seed germination rate is very low, less than 18%, and is usually reproduced only by individual growth through laparotomy. Moreover, because the plant grass is less competitive with other plants and cannot expand its habitat, wild plant grasses cannot supply enough raw materials, and until now, there are almost no research on the plant grass and the development of cultivation technology. All of them rely solely on income.

Therefore, the present invention is to produce a seedlings, including the roots, stems and leaves of the plant artificially aseptically in large quantities in the cabin by using biotechnological technology of the centrifugal plant which is difficult to extract in domestic native areas and dependent on foreign imports. After transplanting seedlings to the purified soil and purifying them, transplant the purified seedlings on land and cultivate them, but harvest them at the time of the maximum accumulation of useful substances in the cultivated plants to produce high quality therapeutic raw materials. To provide a cultivation method.

In order to achieve the above object, the present invention has to produce aseptically a large amount of seedlings, including roots, stems and leaves of the centella, in-flight, and for this purpose, the whole plant is grown aseptically in a liquid medium. Cultured in a sterile B5 medium to which Ross was added, the cultured medium of the centella was used a glass or plastic air-injectable bioreactor or culture bottle that can transmit light.

At this time, when the air flow rate was 0.5 L / min, the temperature was 25 ℃, 16 hours photoperiod and cultured for 8 weeks, the 3 L air-injected bioreactor was able to produce 100 to 120 seedlings. In Esau, 130 to 150 seedlings were produced under the same conditions. Therefore, the growth rate was also faster as the culture vessel is larger.

Next, seedlings of the sterile plants produced by aseptic culture are planted in purified soil mixed with vermiculite and sand in a 1: 1 weight ratio, and the external solar radiation is controlled to 55% or more using a black shading net at a brightness within 15,000 lux. The seedlings were purified and then transplanted to the land to be grown. At this time, it is preferable to set the distance between seedlings to 30-35 cm in consideration of the leaf length and leaf diameter of the stem plants when transplanting the purified seedlings.

At this time, in the transplantation of purified seedlings to the land, the experiments were carried out in consideration of the survival rate of the centella as a result that the preferred temperature is higher than the lowest average temperature of more than 8 ℃ and the average temperature is higher than 13 ℃.

In addition, as a result of investigating the maximum accumulation of useful pharmacological substances in the cultivated plants, the cultivated plants grown on the transplanted lands showed that the average temperature for harvesting after growing for 6 months is the average temperature greater than or equal to 22.3 ℃ and the minimum average temperature. Most preferred was greater than or equal to 17.6 ° C.

The present invention will be described in more detail with reference to the following Examples. At this time, the present invention is not limited in any way by these examples.

Example 1

10 cultivated plant plants, aseptically cultured in a liquid medium, were incubated in sterile B5 medium containing 3% squarose, and the containers containing the medium were light-transparent 3L and 5L air bubbles. type bioreactor was used. At this time, the air outflow rate was 0.5 L / min, temperature was incubated for 8 weeks in 25 ℃, 16 hours photoperiod.

The 3L air-injected bioreactor was able to produce 100-120 seedlings, and the 5L air-injected bioreactor produced 130-150 seedlings under the same conditions.

Example 2

In order to determine the shading level in purifying the seedling seedlings produced by Example 1, seedlings were planted in a purified soil mixed with vermiculite and sand in a 1: 1 volume ratio, and the seedlings were purified with different shading ratios. At this time, the shading ratio was shaded by black shading nets of 0%, 35%, 55%, and 75%, respectively, and the survival rate, plant height, and the presence of new breeding grounds were investigated under low light conditions for 21 days. The results of the experiments are shown in Table 1 below.

As shown in Table 1, in the shading zone, the survival rate is less than 50% and the external solar radiation exceeding 22,000 lux greatly decreases the survival rate, but when the external solar radiation adjusts the shading ratio to 55% or more within 15,000 lux, It forms a new breeding ground and shows good survival rate and plant growth. Therefore, more than 55% of the shading ratio, which can maintain the brightness within 15,000 lux during seedlings, was favorable for the growth of the plant.

Example 3

Purified seedlings are divided into fields and plastic houses from March 21 to April 20 in order to determine an appropriate transplantation time for transplanting the seedlings purified by the second embodiment to the land (field) to be grown. Three weeks after transplantation, the transplantation was performed. The optimal time for transplantation was counted by counting the number of new laparoscopic cells, and the results of the seedling survival rates at the time and place of transplantation are shown in Table 2 below. Indicated.

As shown in Table 2, house cultivation showed a survival rate of 98% or more regardless of the transplant time, and is not limited by the transplant time. In land (field) cultivation, the seedlings transplanted between March 21 and March 31 were used. The survival rate of seedlings transplanted between April 1st-April 10th and April 11th-April 20th was over 98%, respectively. Most transplanted seedlings can survive in April. have. Therefore, most of the seedlings will be viable at the lowest average temperature above 8 ° C and above average 13 ° C (lowest average temperature> 8 ° C, average temperature> 13 ° C). Individuals transplanted between -31 March were most suitable for transplantation between April 1 and April 10 due to slow growth.

Example 4

In order to investigate the implantation distance for increasing the growth rate, the present invention has a spacing of at least 15-20 cm (1 x 1 m2), 30-35 cm (1.5 x 1.5 m2), and 40-45 cm (2 x 2 m2). Ten seedlings were transplanted to each experimental packaging and the growth rate was investigated after 10 weeks in the shading net facility, and the experimental results of the growth characteristics of the seedlings by planting distance were shown in Table 3.

As shown in Table 3, when the interval between the seedling and the description is transplanted in the range of 15 to 20 cm, the occupancy range per seedling exceeds the transplant interval, and as a result, the length of the leaf disease is shown. In the case of transplanting the spacing between the seedlings and the description in the range of 40 ~ 45cm, the occupancy range of the seedlings is narrower than the transplantation interval, and the length of the leaf disease and the leaf diameter are smaller. Therefore, the optimum spacing range of seedling transplantation is almost the same as the occupancy range and the spacing interval of seedlings, and the most desirable raw weight is 30 ~ 35cm.

Example 5

In order to efficiently produce useful materials of Madecassoside and Asiaticoside in the centella, we investigated the change of contents at each time after transplantation. After transplanting to land on March 1, 2001, the leaves and stems of the centennial plants were collected by mid-month from April to October. For sample preparation for HPLC analysis, 100 mg of crushed dry powder was added to 5 mL of 70% ethanol and sonicated for 20 minutes, and the extract was filtered through Whatman (No. 1, 70 mm Φ) filter paper and then rotated. Concentrated on evaporator. The concentrate was redissolved in 1 mL methanol (methanol), 10 μL of which was used as an HPLC sample. The experimental results for the change of useful substance content (mg / g dry weight) in the centella after transplantation are shown in Table 4 below.

As shown in Table 4, the useful substances in the bottle pool are increasing until June, and decreases sharply in July and starts to increase again, indicating the maximum content in the middle of September. In addition, the content decreases again from October, when the average temperature is greater than or equal to 22.3 ° C and the minimum average temperature is greater than or equal to 17.6 ° C (average temperature ≥ 22.3 ° C, minimum average temperature ≥ 17.6 ° C). It is believed that harvesting leaves and stems of the centellae in mid-September with more than 200 hours of sunshine hours per month.

As such, the present invention is a method for mass cultivation of cultivated plants in house or field by investigating shading during cultivation and purifying seedlings optimized for mass production of cultivated plants, transplanting distance between seedlings when transplanting soil, and transplanting time of seedlings. In particular, it is a very useful invention that can contribute to the replacement of income as well as to increase the profits of farms by harvesting the plant plants at the maximum accumulation condition and the optimum time of the useful material in the plant pool.

Claims (4)

Aseptically cultivating the seedling plant seedlings including roots, stems and leaves in the cabin, transferring the cultured seedlings to purified soil, and purifying the purified seedlings with a minimum average temperature higher than 8 ° C. Transplanting the land with the average temperature higher than 13 ℃, and the leafy plants transplanted on the land for 6 months, with the average temperature greater than or equal to 22.3 ℃ and the minimum average temperature greater than or equal to 17.6 ℃ Method of cultivating the centellar plant comprising the step of harvesting the stem. The method of cultivating cultivated plants of claim 1, wherein the cultivation of the cultured seedlings is controlled to 55% or more. The method of claim 1, wherein the purified soil is a cultivation method of centella plant, characterized in that made by mixing the vermiculite and sand in a 1: 1 volume ratio. The method of cultivating a cultivated plant of claim 1, wherein the seedlings and the descriptions are adjusted to 30 to 35 cm in transplanting the seedlings to the land.