CN1016839B

CN1016839B - Method for culturing bupleurum plants and apparatus thereof

Info

Publication number: CN1016839B
Application number: CN88108714A
Authority: CN
Inventors: 渡边齐; 武田继之助; 大盐春治
Original assignee: Takeda Chemical Industries Ltd
Current assignee: Takeda Pharmaceutical Co Ltd
Priority date: 1987-12-18
Filing date: 1988-12-17
Publication date: 1992-06-03
Also published as: JPH01160440A; KR890009245A; CN1033547A

Abstract

To culture the subject plant containing a useful component such as anti-inflammatory agent, promoting quick growth of the plant in a short time, enabling the year-round culture and accumulating a large quantity of saikosaponins in the plant body, by subjecting a plant of genus Bupleurum to nutriculture.The objective plant can be cultured by the nutriculture of a plant of genus Bupleurum. The nutriculture is preferably rock wool culture or hydroponic culture. Preferably, the culture is carried out by using a culture tank lined with a black resin film.

Description

Method for culturing bupleurum plants and apparatus thereof

The present invention relates to cultivate the new method of radix bupleuri plant, the complete stool of this plant, particularly its leaf can be used as the raw material of the saikoside that the tool physiologically active is provided; Also relate to and contain this plant as the antiinflammatory agent of active principle and cultivate the equipment that this plant species is suitable for.

Film edge radix bupleuri (Bupleurum falcatum L.) or the root of drying that belongs to some close plant of (Umbelliferae) together are called as " Bupleuri Radix ", and in many preparatons, used as antipyretic, for example " Shosaikoto ", " Dais ikoto ", " Saikokeishito " or the like, and wild and tame plant all has been used as raw material.Have found that, contain certain oligosaccharides glycosides in the radix bupleuri plant, be referred to as saikoside (abbreviating " S " hereinafter sometimes as), its key component is S.a, c and d.

In recent years, people have noticed that saikoside and derivative thereof are the antiinflammatory agents for the treatment of illnesss such as rheumatism, have now made their physiologically active clear, and are for example active to arthritic auxiliary inhibition.So, use the radix bupleuri plant as the raw material of supply saikoside among constantly increasing (Japanese Patent Publication No.62-186463).

Aspect in addition, though saikoside does not exist only in the radix bupleuri plant roots, also be present in its acrial part, but the radix bupleuri plant of being cultivated has up to now just utilized their root, still found no acrial part that the people utilizes them as the raw material of producing antiinflammatory agent or saikoside.

But for for the radix bupleuri plant of big Tanaka's cultivation, in the relative dry weight of the each several part in its steepest vegetative period, for example leaf accounts for 46%, stem accounts for 40%, and root only accounts for 14%.Aspect in addition, the saikoside content of each several part is: root is about 1.0%, and leaf portion is about 0.6%, stem about 0.05%.Therefore, the saponin content ratio of each several part is a leaf: stem: root=27: 2: 14, so contained saikoside is maximum in the leaf.In addition, wherein the content ratio of S.a, c and d is also different at each several part.For example at root, the amount of S.a and S.d much at one, and the amount of S.c is S.a half.In leaf, the content of S.d is maximum, and the content of S.a and S.c is respectively its about 20-40%.Therefore, the leaf of having found the radix bupleuri plant is suitable for as S.d supply of raw material source most.

But as above-mentioned, the radix bupleuri plant of being cultivated so far all is to be purpose to gather in the crops its root, so the conventional cultivation method all is to focus on to allow root grow up.Therefore, when when gathering in the crops its leaf and be purpose, culture technique that need be different with the cultivation method of routine.

In general, when in the greenhouse that is controling environment during soil cultivation radix bupleuri plant, root not only, acrial part is also grown very fast, therefore can boost productivity greatly, but with when the plant of big Tanaka's cultivation is compared, the former saikoside content is much lower.Because this kind trend is the part especially severe on the ground, so this provides the favorable method of radix bupleuri blade hardly.

The present inventor is that deep research has been carried out on the basis with the above-mentioned fact, and its objective is provides a kind of improved cultivation method, makes it to be suitable for producing the leaf of the radix bupleuri plant that has the high-load saikoside in a large number.Result of study is found, adopts nutrition cultivation (soilless culture) to suit.

So one of purpose of the present invention provides a kind of method that is suitable for producing the radix bupleuri plant leaf blade.

Two of purpose of the present invention provides a kind of antiinflammatory agent, contains in this medicine by method gained plant of the present invention as active principle.

Three of purpose of the present invention provides a kind of plant cultivation equipment of the present invention of implementing that is applicable to.

By the following description and accompanying drawing, those skilled in the art can understand above-mentioned purpose of the present invention and other purposes and superiority thereof.

Shown in Figure 1 is mineral wool cultivating device schematic diagram, is the system that nutrient solution is added drop-wise to mineral wool cultivation bed shown in it.

Shown in Figure 2 is the water planting schematic diagram, wherein shows the system of batch (-type) supply nutrient solution.

Fig. 3 and 4 is respectively the side generalized section of each embodiment of present device.

So one aspect of the present invention is a kind of method of cultivating the radix bupleuri plant, it is characterized in that this radix bupleuri plant is to adopt nutrition cultivation gained.

Another aspect of the present invention is a kind of anti-inflammatory agent, and the leaf that comprises a Plants in this medicine is as active principle, and this plant is to be got by the nutrition cultivation by a kind of radix bupleuri plant, and contains the saponin of the amount of increasing.

A further aspect of the invention is exactly that a kind of plant nutrient cultivation uses equipment, this equipment to be applicable to this kind nutrition planting type, is specially adapted to water culture, and the inner surface that it is characterized in that the cultivation bed of this equipment is black. As an a kind of example of plant nutrient cultivating device, comprising having:

A cultivation bed, the inner surface of this groove is lined with one deck black resin film, also is fixed with a drainpipe in its bottom, in order to drain nutrient solution;

A cultivation case is positioned at the top of described cultivating groove, just be stacked in it above, can pack into one in the described cultivation case and cultivate bed, the cultivation bed is provided with many openings, allows root pass therethrough;

A nutrient solution basin is arranged at the bottom of described cultivating groove, wherein also serves as a contrast with black resin film, and it is connected with the drainpipe of described cultivating groove;

Nutrient solution is delivered to the used equipment of described cultivating groove from the described nutrient tank follow procedure stipulated time.

In recent years, horticulture person extensively adopts the nutrition cultivation as a kind of means that prevent the destruction of continuous farming, disease or insect pest.This kind cultivation method is characterised in that by manual control environment and nutritional condition.Known that above-mentioned feature can prevent above-mentioned shortcoming, and can promote the growth of plant greatly.But nutrition cultivation up to now mainly is the occasion that need to be used to the plant shoot branch in practicality, becomes bed course shape rootlet because the method trends towards tachyauxesis, and is not suitable for the growth root.Also be out of use in the main medicinal plant that uses root of cultivation, even people think that so far the nutrition cultivation is not suitable for the cultivation medicinal plant.In addition, as mentioned above, though the radix bupleuri plant promotes the growth of its acrial part under the controlled environment condition when earth culture, compare with institute growing plants in natural environment as mentioned above, saikoside content wherein is much lower.Yet, be surprised to find that in the present invention, can gather in the crops the radix bupleuri plant leaf blade that saponin content increases by the nutrition cultivation.In addition, radix bupleuri is perennial plant in Japan.Generally speaking, seed germinateed in spring, and acrial part is in death in autumn (October is to December), but root lives and passes the winter.So to spring next year, this plant grows sprouting again again.But by the present invention, it is possible that this plant species is carried out annual cultivation.In addition, can also gather in the crops simple and efficiently and contain the higher radix bupleuri plant of saikoside amount in the leaf.Therefore can expect to boost productivity greatly, and this is epoch-making aspect medicinal plant cultivation.

So, by cultivation method of the present invention, except that being applicable to Bupleurum falcatum L., be applicable to that also other contain the radix bupleuri plant of saikoside, for example Bupleurum falcatum var.kiusianum, Bupleurum Chinese (B.Chinese), radix bupleuri scorzoneraefolii (B.scorzonerae folium), B.dahurium, B.euphorbioides, B.jeholense, B.marginatum or the like.

About nutrition cultivation system aspect, the known system that solid is arranged, for example sand culture, gravel culture, charcoal cultivation, polyurethane cultivation and mineral wool cultivation also have non-solid system, for example water planting, steam cultivation and spray culture.In the present invention, can use any of above-mentioned these systems.But used system should be able to fine draining when cultivation radix bupleuri plant, preferably adopts mineral wool cultivation or water planting, because draining easily.

In general, nutrition cultivation of the present invention is carried out in the controlled environment condition, for example, the greenhouse that can adopt materials such as glass, polyacrylate or polyvinyl chloride to build, and can pass through temperature in the thermal source control greenhouses such as electricity, combustion gas, kerosene, planting seed in suitable cultivation bed, is supplied with a kind of nutrient solution culture and brought out rice shoot, gather in the crops these plants then.

Seed used among the present invention is not particularly limited, and any available radix bupleuri plant seed all can use.And the cultivation bed of sowing usefulness also is not particularly limited, be generally easy to operate for the purpose of, preferably use the mineral wool piece or the mineral wool bed course of appropriate size.Mineral wool is a kind of of inorfil, is to be subjected to about 1600 ℃ of fibers that high temperature is made by basalt, slag etc. for raw material.The composition of the mineral wool of using on the agricultural comprises fiber (3-4%) and porous solids, it is moldedly become bulk or bed course shape, and make it to have water imbibition and be adjusted to certain pH value, and this material has commercially available product, can be applied to the present invention.More specifically say, found to use separately brightness edge rock to be applicable to cultivation radix bupleuri plant, so as preferred material of the present invention as the made mineral wool piece of raw material or the chemistry and the physical property of bed course (for example, producing) by Dutch Gloudan Co..

In the present invention, preferred condition is that seed is maintained at about 18-22 ℃, is generally 20 ℃, under daylight (about 5-20 hour) or in the dark, till germinateing fully, only supplies with water during this section.Generally speaking, irrigate 1-5 time, be generally once a day, in case seed becomes dry with enough water.After this, seed sprouting in about 30 days.

Then, supply with nutrient solution to the cultivation bed and carry out the nutrition cultivation.About used nutrient solution, can use the nitrogenous source solution that contains the nitrate form, for example use potassium nitrate, nitrate of lime or the like, and/or use the salt of ammonia, for example ammonium phosphate, diammonium phosphate or the like; Phosphorus source: for example ammonium phosphate, diammonium phosphate, phosphoric acid or the like; Potassium source: potassium nitrate for example; Calcium source: for example phosphoric acid-calcium, calcium chloride or the like; Magnesium source: magnesium sulfate for example; Also has micro-source: iron, for example chelated iron (ethylenediamine tetra-acetic acid iron); Boric acid; Manganese chloride; Zinc chloride; Copper sulphate; Ammonium molybdate or the like.Usually, the working concentration of these compositions, nitrogen be 2.5-20 molar equivalent (me)/liter, best 9.3me/ liter (being equivalent to 130ppm); Phosphorus is the 1-6me/ liter, preferably 2.6me/ liter (being equivalent to 60ppm); Potassium is the 1-10me/ liter, preferably 3.8me/ liter (being equivalent to 180ppm); Calcium is the 1-10me/ liter, preferably 4.1me/ liter (being equivalent to 115ppm); Magnesium is the 0.5-5me/ liter, preferably 1.8me/ liter (being equivalent to 37ppm).This kind nutrient solution has commercially available product, Otsuka House No.1 and No.2 that for example Japanese Otsuka Chemical Co. sells.This solution can use a half strength of its standard volume in use.

It should be noted that especially that when with some organic acid when for example malonic acid, acetate or the like or their alkali metal salt (for example sodium salt etc.) or ammonium salt joined in the described nutrient solution (the about 50-500 mg/litre of addition), the saponin content in the plant increased.So it is favourable adding some organic acid in this nutrient solution.

In the present invention, during the nutrition cultivation, optimum condition is to be maintained at about 20-35 ℃ of temperature, is about 25 ℃ usually, and lasts about 14-20 hour under daylight.During this period, the time interval of taking follow procedure with suitable flow this nutrient solution is dripped or batch (-type) adds, and for example adds fashionablely at batch (-type), can add once in per 10 minutes to per 4 hours, and normally each hour adding once.Preferably utilize the electrical conductivity (EC) of nutrient solution to control its concentration, making its seedling stage after germination is 0.3-0.8 milli Siemens/cm, be generally 0.5 milli Siemens/cm, the vegetative stage thereafter is a 0.5-4.0 milli Siemens/cm, is generally 1.0 milli Siemens/cm.After this can impel its ramp, concurrent light soy sauce tongue.Usually, when summer culture, can gather in the crops the high leaf of saponin content in after planting three months, when cultivating in the winter time, can gather in the crops in after planting four months, cultivation 3-5 time continuously within the same set of equipment 1 year.In addition, the equipment of application multiple field also can further improve yield per unit area.

Described in detail by hereinafter reaching the superior embodiment of accompanying drawing nutrition cultivation system of the present invention.

Fig. 1 is the schematic diagram of mineral wool cultivation, wherein is provided with the feed system that drips nutrient solution.In system shown in Figure 1, be that (this mineral wool piece is of a size of: 3.5-10.0 centimetre cube the upper face of the planting seed of radix bupleuri plant in mineral wool piece (1); Highly be 4.0-10.0 centimetre; Commonly used is 10.0 * 10.0 * 8.0 centimetres), and these pieces are placed on mineral wool batten (2) go up and (be of a size of: width 15.0-60.0 centimetre; Length 90.0-120.0 centimetre; 0.8-7.5 centimetre of height; Commonly used is 30.0 * 90.0 * 7.5 centimetres).Going up the water of irrigating capacity to piece (1) became dry before germinateing to prevent seed.50-1000 milliliter (usual amounts is 100 milliliters) nutrient solution is delivered to these mineral wool pieces (1), solution is that (this tank volume is the 50-2000 liter among the nutrient tank (4) of storing in independent setting, commonly used is 300 liters), utilize pump (6) to enter by feed tube (7), wherein there is controller (5) to come the feeding time, concentration of control pump and gives liquid measure, the nutrient solution flow is the 5-50 ml/min, 10 ml/min normally, and by time interval feeding given in advance.Too much nutrient solution flows out from the bottom of mineral wool batten (2).With natural conditions oxygen supply gas in the cultivation bed, therefore do not need to consider to use the artificial ventilation by the hole in the mineral wool.

Shown in Figure 2 is the schematic diagram of a cover hydroponic system, and this system has the equipment that batch (-type) is supplied with nutrient solution.In system shown in Figure 2, the sowing of radix bupleuri plant seed upward (is of a size of: width 15.0-60.0 centimetre at a thin mineral wool bed course (1), length 50.0-120.0 centimetre, 0.8-5.0 centimetre of height, Commonly Used Size is 28.0 * 58.0 * 1.8 centimetres), and this bed course placed on the cultivating groove (3) (be of a size of: width 30.0-150.0 centimetre, length 50.0-300.0 centimetre, degree of depth 3.0-30.0 centimetre, Commonly Used Size is 60.0 * 180.0 * 5.0 centimetres).Commercially available have a kind of plastic box of cultivating rice seedlings to be applicable to very much this bed course of support.Go up the water of irrigating capacity at mineral wool bed course (1), in case seed became dry before germinateing.Nutrient tank (4) (volume 50-2000 liter from independent setting, 300 liters of volumes commonly used) utilize pump (6) that nutrient solution is delivered to cultivating groove (3) by feeding pipe (7), on pump, connect a controller (5), in order to control feeding time, concentration and to give liquid measure, the nutrient solution flow is the 20-100 ml/min, usual amounts is 70 ml/min, and by time interval feeding given in advance (per hour being generally once).This plant just grows by being dipped in the rootlet in the liquid, and rootlet passes mineral wool bed course (1) contact nutrient solution.Make nutrient solution in cultivating groove (3), retain certain hour by means of valve-off (8), flow into nutrient tank (4) by the outage (9) that is arranged at cultivating groove (3) bottom then.Be full of a period of time before the nutrient solution once more at cultivating groove (3), these rootlets are exposed in the air and obtain oxygen supply.

Use this system, promoted the growth of leaf greatly.For example, when cultivation in mineral wool in the time of 3-4 month, never the leaf harvest yield that obtains of bolting plant is equally matched with the leaf harvest yield of cultivating about 9 months plant in back big Tanaka.In addition, promptly begin bolting by plant that the present invention cultivated in stage early, so be expected to increase harvest yield.For example, when adopting the mineral wool cultivation, the ratio of bolting plant reaches about 30% in four months, and when conventional field production, the bolting plant ratio of same time is no more than 20%.In addition, when adopting the mineral wool cultivation, the saikoside content in the leaf significantly increases.

After cultivation is finished, can gather in by known method, for example, the root that will pass the cultivation bed earlier cuts off, and then acrial part is pulled out from the cultivation bed.

The plant of having gathered in is pressed root, stem and leaf separately, carry out drying then, for example 40-80 ℃ of drying, be preferably 60 ℃, be 24-72 hour drying time, and commonly used is 48 hours, the for example available electric dry oven of drying source is used as them antiinflammatory agent then or sees the raw material of saikoside as.

Antiinflammatory agent of the present invention contains the radix bupleuri plant of as above gained as a kind of active principle.The cured leaf original can be broken into powder and promptly be used as this kind antiinflammatory agent, perhaps with after the solvent extraction with the extract form as this kind antiinflammatory agent, solvent for use can be water, alcohol, for example methyl alcohol, ethanol or the like or their mixture, extraction temperature is 10-100 ℃.They can be prepared becomes pulvis, granule, fine grained agent or the like for oral, or make ointment, opoultice or the like for parenteral administration, or make medicinal tea, fry in shallow oil decoction or the like, wherein can use pharmaceutically acceptable excipient, as talcum powder, alumina silicate or the like, and by producing upward known method use.

This kind active principle of the present invention is to be used as antiinflammatory agent, is 2-400 milligram/day (S.d, oral), perhaps parenteral administration 1-200 milligram to the required dosage of being grown up.Saikoside itself is so far extensively as the component of Bupleuri Radix, and fully determined their hypotoxicity and safety in practicality.Therefore, antiinflammatory agent of the present invention has shown excellent anti-inflammatory activity, and is applicable to the treatment rheumatism.

As previously mentioned, the present invention also provides the equipment that equipment of the present invention, particularly water planting are used of implementing that is applicable to.In its preferred embodiment shown in Fig. 3 and Fig. 4.

Equipment shown in Figure 3 comprises cultivating groove (3) (635 * 290 * 120 millimeters of the inside dimensions of being made by polystyrene foam plastics, 23 millimeters of thickness), inner face lining is with black and white bilayer film (Japan, Mikado Kako Co., Ltd. the polyolefin of Zhi Zaoing), black is spread (not shown) outwardly, also comprise cultivation case (10) (rice seedlings incubator, 580 * 280 * 20 millimeters of inside dimensions) be stacked and placed on this cultivating groove top, nutrition liquid bath (4) (660 * 100 * 200 millimeters of the inside dimensions that also comprise polystyrene foam plastics system, 33 millimeters of thickness), place under the cultivating groove (3), and with the cultivation case the same liner of spreading.In this planting type, meeting is grown the water silk floss and is produced the problem that hinders normal running in nutrient solution.But, have been found that equipment surface becomes black, can prevent the influence of sealing silk floss, for example available black cloth, blacking, black paper, black glass, black plastic film or the like.Therefore, in equipment of the present invention, not only store the cultivating groove of nutrient solution, also will make black (for example using black plastic film) where necessary together with the inner surface of nutrient solution storage tank.Where necessary, the also available black material of other parts hides (for example using black plastic film or the like).

As shown in Figure 3, this cultivation bed (mineral wool bed course, 500 * 280 * 290 millimeters) is to be contained among the cultivation case (10), and plant is growth in this cultivation bed just, opening is arranged at the bottom of bed, makes the root of plant can pass the cultivation bed and stretches in the nutrition liquid layer of cultivating groove (3).The position of these openings, size and number can suitably be arranged by number and the shape of plant.When the cultivation case is when being made by polystyrene foam plastics, opening these mouths in the above is easily.

Drainpipe (9) (13 millimeters of external diameters are made by polyvinyl chloride) is fitted in the bottom of cultivating groove (3).By regulating the height of drainpipe, can make the nutrient solution in the cultivating groove (3) remain on desired depth, if an opening (not shown) is set again, emit with regard to being easy in the cultivating groove (3) too much nutrient solution in its bottom.The bottom of drainpipe (9) feeds nutrition liquid bath (4).

By pipeline (11) pump (6) is connected to nutrition liquid bath (4), by feed tube (7) nutrient input is delivered to cultivating groove (3) by pump (6).Come control pump (6) by controller (5) (a for example available timer).The logistics of forming nutrient solution by controller (5), pump (6), feed tube (7) and pipeline (11).

Equipment shown in Fig. 4 is the modification of equipment shown in Figure 3, and it is substantially the same with Fig. 3, and difference does not just have pipeline (11) and the pump (6) among Fig. 3, but replaces with the pump of formula under water in the nutrition liquid bath.

When application equipment of the present invention is cultivated, nutrient solution in the nutrition liquid bath (4) is sent into cultivating groove (3) by pump (6) and feed tube (7) at interval with preset time, for example, from per 2 hours of o'clock at 8 o'clock to 18 liquor charging 10 minutes, from per 4 hours of o'clock at 18 o'clock to 8 liquor charging 10 minutes, make the root of stretching from cultivation case (10) bottom be immersed in the nutrient solution.

Equipment of the present invention can be with the material manufacturing that is easy to get, and is easy to assembling and carries out miniaturization.So this equipment is specially adapted to cultivate the radix bupleuri plant.But the purposes of this kind equipment is not limited to this, can also be used to cultivating other each plant species.

The following further sets forth in detail of example cultivation method of the present invention and antiinflammatory agent, but these examples are not limiting the scope of the invention.

In following example, the analytical method of saikoside is as follows:

The plant of gathering in the crops is divided into root, stem and leaf part, and respectively 60 ℃ of dryings 48 hours.Grind the powder that becomes 100 sieve meshes then respectively, as test specimen.

(HPLC) measures its saponin with high performance liquid chromatography.Concrete grammar is as follows: accurately take by weighing sample (0.2 gram), to wherein adding 9 milliliter of 80% methyl alcohol and 0.2 milliliter of ammoniacal liquor.Then the gained mixture was handled 1 hour with ultrasonic.Adding 80% methyl alcohol in gained suspension makes its cumulative volume reach 10 milliliters (exact values).Clear liquid takes out two parts of samples of 0.5 milliliter (exact value) from it, respectively to wherein adding entry (5 milliliters).They are sent in two small chromatographic columns (U.S., J.T.Baker Corp. makes) respectively and purify, after by 5 milliliter of 30% methyl alcohol, again with 10 ml waters by this post, one of them post is with methyl alcohol (4 milliliters) wash-outs (A).

Water in another post is with 40% FeCl ₃The aqueous solution replaces, and stops 20 minutes, and the epoxide ring of S.a, c and d is opened, and obtains S.b ₁H and b ₂In this post, feed 10 ml waters then, in order to wash-out FeCl ₃, use the further wash-out saponin of 4 ml methanol (B) then.Add methyl alcohol respectively to eluate (A) with (B): ammoniacal liquor (97: 3) mixed liquor is adjusted to 5 milliliters (exact values) with its volume.Its 10-20 microlitre is carried out HPLC as sample by following condition to be analyzed.If need, carry out earlier solution being filtered through millipore filter before HPLC analyzes.

The efficient liquid phase chromatographic analysis condition

Detector: the bavin outside line monitor of variable wavelength (Japan, Shimadzu Seisaku-sho makes, the C-R2AX type)

Fixing phase: 6 * 200 millimeters of Elmer C-18(Φ)

Flowing phase: methyl alcohol: water (76: 24), 1 ml/min.

Detect wavelength: 251 millimicrons

Utilize ready-made in advance calibration curve, according to by each peak area of solution A gained: S.b ₁(HPLC retention time: 21.5 minutes), S.h(retention time: 7.8 minutes) and S.b ₂(retention time: 15.1 minutes) can determine their content separately.Same the method determines each content of solution B.Poor according to each content among solution A and the B can calculate the content of saikoside a, c and d.

Example 1

Adopt nutrient solution to drip the mineral wool cultivation of system

Film edge radix bupleuri (Bupleurum falcatum L.) planting seed upward and with this piece is placed glass box at mineral wool piece (10 * 10 * 5 centimetres), and the temperature inside the box is about 20 ℃.In order to prevent that seed from becoming dry, irrigate nozzle trickle 1 minute by one every day.After 37 days, the plant of these leaves that begin to grow transferred to together with the mineral wool piece the cotton lath of a lump ore (90 * 30 * 7.5 centimetres) is gone up and the greenhouse of the polyacrylate system of placing, indoor temperature remains at 25-35 ℃ of scope.The method of preparation nutrient solution is with Otsuka House No.1 and No.2(Japan, Otsuka Chemical Co.Ltd produces) make the 75ppm and the 50ppm aqueous solution respectively, then two kinds of preparing solutions are made its electrical conductivity (EC) reach 1.25 milli Siemens/cm (pH=5.5-6.5).Select every day from the morning 5 to per 1 hour of 6 pm and send into once this nutrient solution automatically, last 10 minutes.Lifting rate is adjusted to 10 ml/min.

After planting 3 months, leaf taken down and in 60 ℃ of dryings 48 hours.Pulverize with grinder then, become 100 sieve mesh powder.Measure its saponin content then.In the result of gained shown in the table 1.

Group is also listed in the analysis result that the plant of being cultivated with big Tanaka in the glass greenhouse soil obtains in the table 1 in contrast.Concrete condition is in glass greenhouse, broadcasts into seed when using the soil (1/2000 acre) that is contained in the Wagner basin to cultivate with above-mentioned mineral wool.After the sowing, per two days trickles are (200 milliliters) once.After the germination, when leaf began to grow, spray Hyponex(Hyponex Co. produced powder) aqueous solution of 1000 times of dilutions, jede Woche drenches 2 times, and every basin is with 500 milliliters.During the cultivation, temperature remains on 15-35 ℃.After planting 3 months, receive its leaf, measure its saponin content by the same way as of above-mentioned mineral wool (being called RW hereinafter) cultivation gained leaf.

About the plant of field production, (spring), (Fukuchiyama-shi Kyoto-fu) sowed for Japan, Osada, and results obtain this plant after 5 months the Tanaka of 1289-1 black volcanic ash clay loam in above-mentioned mineral wool cultivation.About the fertilising situation, per 10 acres of Tanakas, use dried poultry manure (300 kilograms), calcium hydroxide (100 kilograms) and a kind of mixed fertilizer Rinkaan15 number (Japan, Nissan Chemical Industry Co. produces, wherein N: P: the K composition ratio is 15: 15: 15) (20 kilograms) as base manure.At after planting 4 months, use the same mixed fertilizer as topdressing.Seeding quantity is 2 liters/10 acres, adopts drilling method, 15 centimetres of line-spacings, 80 centimetres of spacing in the rows.Sow seeds and blinding after, 500 times of water diluents (100 liters/10 acres) of spray Trefanoside emulsion (Ltd. produces for Japan, Takeda Chemical Industries Co.).By measure its saponin content with previous mode.

Shown in the table 1 about the data of leaf.The result is, adopts the mineral wool cultivation and adopt soil cultivation in the polyacrylate greenhouse in glass greenhouse, compares with the crop of field production, and the above two grow very rapid in a short-term.Its prominent features is that the leaf of bolting plant is not many than the plant leaf recuperation of field production.In addition, about the harvest yield of leaf, the leaf amount of field production accounts for about 46% of plant gross weight, and adopt the mineral wool cultivation reach 68%.About the dried harvest yield of product, low slightly during than field production when cultivating with mineral wool.

In the product of glass greenhouse earth culture, saponin content is more much lower than the field production, but the content that obtains in the product of mineral wool cultivation is much higher.Especially S.d content is more much higher than the content of S.a and S.c.As a result, great changes have taken place because of used cultivation method for the saponin total content of each strain plant, as shown in table 1, and the content of the product of mineral wool cultivation is equally matched with the root of field crop gained biennial plant.

Table 1

Saponin content in the growth of film edge radix bupleuri (Bupleurum falcatum L.) and the blade thereof

Cultivation land for growing field crops, polyacrylate garden glass greenhouse, place

Cultivation medium RW soil soil

Bolting rate (%) 4.0 3.8 14.1

Leaf number 5.3 5.1 6.4

Heavy (gram) 2.2 of bright leaf ± 0.5 2.1 ± 1.1 2.3 ± 2.1

Heavy (gram) 0.55 0.54 0.61 of cured leaf

Do utilization percent (%) 25.1 25.8 26.6

Saponin content

(milligram/strain plant)

S.a 1.41 0.36 0.49

S.c 1.72 0.56 0.85

S.d 5.79 1.57 1.77

Example 2

The mineral wool cultivation of Bupleurum falcatum var.kiusianum

According to the same way as of cultivation film edge radix bupleuri (Bupleurum falcatum L.) described in the example 1, in the polyacrylate greenhouse, carry out mineral wool cultivation, carry out earth culture big Tanaka, difference is that the seeding quantity big Tanaka is 2.6 liters/acre.Data at the leaf that produces shown in the table 2.

In the mineral wool cultivation, similar with the situation of film edge radix bupleuri (Bupleurum falcatum L.), to compare with field production, plant growing is healthy and strong, and growth is more quick at short notice.Can see that by table 2 compared with field production by the leaf of mineral wool cultivation gained, saponin content is much higher.But, because the saponin content of the acrial part of Bupleurum falcatum var.kiusianum is original just very low, so the saponin content of its every strain plant will be lower than every strain content of film edge radix bupleuri (Bupleurum falcatum L.).

Table 2

Saponin content in the growth of Bupleurum falcatum var.kiusianum and the leaf thereof.

Cultivation land for growing field crops, polyacrylate greenhouse, place

Cultivation medium RW soil

Bolting rate (%) 3.1 7.2

Leaf number 4.9 6.1

Heavy (gram) 1.7 of bright leaf ± 0.5 1.8 ± 1.4

Heavy (gram) 0.39 0.44 of cured leaf

Dry labor rate (%) 23.6 24.9

Saponin content (milligram/strain plant)

S.a 0.15 0.13

S.c 0.12 0.09

S.d 0.70 0.48

Example 3

Applied nutrition liquid is the water planting of watering system intermittently

Placing the seed of broadcasting on the mineral wool bed course of rice seedlings incubator (58 * 58 * 2 centimetres) with film edge radix bupleuri (Bupleurum falcatum L.), place glass greenhouse then, indoor temperature is about 20 ℃.After 33 days, the plant that begins frondesce transferred to together with incubator (polystyrene foam plastics system, 50 * 112 * 5 centimetres) indoor temperature remains on 25-35 ℃ in the cultivating groove in the polyacrylate greenhouse, continue cultivating and growing.About nutrient solution, use the aqueous solution of 1000 times of Hyponex dilutions, and then regulate its concentration, make that the electrical conductivity (EC) in the seedling stage after just having germinateed is 0.5 milli Siemens/cm.Thereafter 3 months is 1.25 milli Siemens/cm.At 8 in every day from the morning is to 6 pm, makes nutrient solution circulation 10 minutes in per 2 hours; 8 points from 6 pm to the morning, circulation in per 4 hours 10 minutes.

Checked plant growing situation to the in every month till five months (cultivation phase: 4 months), the result of gained shown in the table 3.Bolting rate in 4 months is 27.1%, and similar to above-mentioned example 1, and this value is more much higher than the field production.Aerial growth situation of plant and mineral wool grower also just demonstrate the fast of plant growth fraction field production at short notice much at one.Do the low 3-4% of productivity ratio land for growing field crops gained person.

About the saponin content in the leaf, the situation of its result and the cultivation of above-mentioned mineral wool much at one.That is the content in the biennial plant root of S.d content in the leaf and field production is equally matched.And S.a and S.c content are than the height of field production.

Table 3

Saponin content in hydroponics growing situation and the leaf thereof

2 months 3 months 4 months cultivation phase

Bolting rate (%) 2.8 11.3 27.1

Leaf number 4.9 8.7 11.2

Heavy (gram) 1.9 of bright leaf ± 0.4 5.4 ± 2.5 7.6 ± 3.4

Heavy (gram) 0.49 1.25 1.79 of cured leaf

Dry labor rate (%) 24.8 23.2 23.5

Saponin content

(milligram/strain plant)

S.a 0.218 0.223 0.256

S.c 0.330 0.295 0.312

S.d 0.837 0.945 1.043

Example 4

Example 1 is cultivated cured leaf (100 gram) corase grind of gained film edge radix bupleuri (Bupleurum falcatum L.) in the polyacrylate greenhouse broken and add 500 ml waters.This mixture is decocted, reduce to 250 milliliters until cumulative volume.Filter gained mixed liquor, the decoction that obtains frying in shallow oil with cloth.

As previously discussed, cultivate the radix bupleuri plant with the controlled nutrient solution liquid-supplying system of use under the controlled temperature condition, just might promote ramp at short notice, the product amount that obtains is also more much higher.Meanwhile, can also accomplish annual this plant species of cultivation.In addition, the applied nutrition cultivation just might be gathered a large amount of saikosides in plant.Therefore, the productivity ratio and the profitability of whole strain plant and leaf can improve greatly.Promptly can obtain having the very blade of the radix bupleuri plant of effective thus, and obtain a large amount of saikosides sustainably, the purposes that these raw materials are used to develop medicine is extremely wide.

Claims

1, a kind of method of cultivating the radix bupleuri plant is characterized in that adopting that to have inner surface be that the cultivating device of the cultivating groove of black carries out mineral wool nutrition cultivation with the radix bupleuri plant.

2, implement the described equipment that the radix bupleuri plant is carried out mineral wool nutrition cultivation of claim 1, comprising:

(a) cultivating groove is lined with one deck black plastic film in wherein, and is fixed with one from wherein discharging the drainpipe of nutrient solution in its bottom;

(b) cultivation case, this case places the top of described cultivating groove and thereon stacked, the described cultivation case mineral wool cultivation bed of packing into, and have the opening that the root that makes plant can pass in this bottom;

(c) nutrition liquid bath, this groove place described cultivating groove below, in be lined with black plastic film, and be connected with drainpipe on the described cultivating groove;

(d) nutrient solution follow procedure official hour is delivered to the equipment of the required usefulness of described cultivating groove from described nutrition liquid bath.