CN102499044A - Cultivation method capable of increasing biomass of selfheal clusters and yield of active ingredients - Google Patents
Cultivation method capable of increasing biomass of selfheal clusters and yield of active ingredients Download PDFInfo
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Abstract
The invention provides a cultivation method capable of increasing biomass of selfheal clusters and yield of active ingredients. A proper amount of potassium in a water culture nutrient solution is added under the water culture condition suitable for the growth of selfheal, so the biomass of single selfheal cluster is increased obviously and the yield of the main active ingredients (such as rosmarinic acid, ursolic acid, oleanolic acid, total flavones and water extract) is increased obviously. The water culture method is simple, convenient and scientific in operation, small in area occupation, low in cost and wide in application range. The cultivated selfheal clusters have good commodity characteristics, stable yield, high yield of the active ingredients and can provide high-quality raw medicinal materials for production in medicine and hygiene industry and processing and extracting industry.
Description
One, technical field:
This research relates to a kind of cultivation method that improves selfheal fruit ear biomass and active component output, belongs to traditional Chinese medicine culture technique field.
Two, background technology:
Selfheal Prunella vulgaris L. is used as medicine with dry fruit ear for the Labiatae selfheal belongs to herbaceos perennial, is China's conventional Chinese medicine material.The selfheal flavor is hot, bitter, and is cold.Return liver, gallbladder channel.Have and clear liver and improve vision, the effect of dispersing swelling and dissipating binds; Selfheal is usually used in treating thyromegaly clinically, scrofula, the proliferation of mammary gland, pulmonary tuberculosis, acute icteric catarrhal jaundice, diseases such as hypertension; Modern chemistry research shows that selfheal contains multiple triterpenes, flavonoids, phenolic acid class, polysaccharose, sterols, Coumarins, organic acid, plain type of phenylpropyl alcohol and volatile oil composition.Confirm that according to another pharmacological research selfheal water extraction and flavonoids component (Luteolin and Quercetin) all have antitumor activity; Ursolic acid and oleanolic acid have antibiotic, anti-oxidant, antitumor and hypoglycemic biologically active; Rosmarinic acid can anti-oxidant, anti-inflammatory; In addition, Rosmarinic acid is also by " Chinese pharmacopoeia (2010 editions) is as selfheal quality of medicinal material control index.
Selfheal is as the large medicinal material of China, and market demand is bigger; The domestic and international market demand is about 2000~2500 tons, and wherein 15% for export.In recent years, the As market amount is constantly soaring, and the selfheal wild resource suffers excessively to excavate, and adds environmental disruption, and its stock number is sharply exhausted, can not satisfy medicine and related industry demand; Therefore, at the beginning of the nineties in last century, various places begin the domestication and introduce a fine variety selfheal, guarantee the regular supply of selfheal medicinal material with this; But; Also there are some outstanding shortcomings in the artificial planting selfheal; Long like tradition crop cycle; Plant strain growth is irregular, sowing quantity is big, damage by disease and insect is serious, manual work is wasteful, and residue of pesticide are more, the medicinal material yield and quality forms and is subject to factor affecting such as weather, season and soil environment, can not satisfy the selfheal large-scale planting and produce needs.And soilless culture is than having soil cultivation to have more to many advantage; Compare with earth culture; Water planting has plant convenience for washing, plant article lover, it is easy to change crops, can be in same place annually cultivating, cultivation cycle short, save fertilizer, can effectively avoid characteristics such as continuous cropping obstacle, cropping index height; Therefore, the development of the area of medicinal plant soilless culture in recent years presents fast rise trend.
Current; Many links such as the artificial earth culture plantation in China selfheal main producing region needs to put in order ground, sowing, thinning, weeding, gather; Growth cycle needs about 9 months; Promptly sow mid-October then to mid-June next year and gather, because factor affecting such as long-term continuous cropping and soil-borne diseases, serious damage by disease and insect often appears in selfheal; In addition, external environment is unstable, also makes selfheal when gathering, and its fruit ear maturity is inconsistent, directly influences the selfheal medical material quanlity.The selfheal fruit ear that earth culture is produced, commodity property is relatively poor, and quality is more bad, and output is not high.
At present; The selfheal research work is mainly concentrated on aspects such as germ plasm resource genetic diversity, the textual criticism of this agrostology, culture technique, chemical composition, pharmacologically active, clinical practice both at home and abroad, and the research of how to use water culture raising selfheal fruit ear biomass and active component content thereof is rarely had report.Good, best in quality, the stable yield of selfheal medicinal material exterior of commodity that adopts water culture to produce; Do not receive factor affecting such as season, weather and soil, can realize selfheal stable yields, high-quality, High-efficient Production.
Three, summary of the invention:
1, technical problem
The object of the invention provides a kind of cultivation method that improves selfheal fruit ear biomass and active component output thereof, so that selfheal well-grown under the water planting condition, individual plant fruit ear biomass increases, active component output also improves.
2, technical scheme
The present invention provides a kind of ciltivating process that improves selfheal fruit ear biomass and active component output thereof, and it may further comprise the steps:
(1) collect seed: collect selfheal Prunella vulgaris Linn seed, select seed even, full, no damage by disease and insect, dehydration makes water content be lower than 12%, and sealing is preserved;
(2) presoaking and germinating: in insulating box, carry out, seed is put into magnetized drinking water soak 2~3h, 4~5 times repeatedly, total seed soaking time 12h; The seed soaking condition: temperature is 20 ± 2 ℃, and illumination is 2000~2200lux, handles making the selfheal percentage of seedgermination be higher than 95% through vernalization;
(3) artificial breeding: when 70% plant grows 2 true leaves, be positioned over and adopt cave dish seeding and seedling raising in the husky bed in greenhouse October then, matrix is river sand: vermiculite: peat=1: 1: 1, normal water and fertilizer management;
(4) transplant planting: late Febuary next year is chosen the seedling that grows 6 true leaves to early March, with distilled water root is cleaned, and is put in the water planting case that fills nutrient solution; Medium evenly passes through gas 1h with air pump every day continuously; On the plastic plate of medium plastic containers, be installed with 15 apertures of diameter 3cm, every strain plant is fixed in the aperture, and per 2 young plant water planting spacings are 10cm, and each is handled plastic containers and is positioned under the shade at random;
(5) configuration potassium cellulose content is 6.0mmolL
-1The Hogland culture fluid, the potassium element disposes with potassium chloride (KCl); Every 7d changes culture fluid, and regularly adds distilled water and keep culture fluid concentration and volume; With HCl and NaOH regulator solution pH 6~6.5;
(6) booth water planting case is cultivated: it is 40%~45% black sunshade net that 5m place, water planting plant top covers one deck light transmittance; Strain was cultivated in booth water planting case mid-March next year to mid-June, and take fruit ear during the fruit ear brownish red mid-June; Naturally dry, put aeration-drying place storage.
Particularly, step also can be:
(1) collect seed: will collect selfheal P.vulgaris L. seed, and select seed even, full, no damage by disease and insect, dehydration makes water content be lower than 12%, and sealing is preserved.
(2) presoaking and germinating: the selfheal presprouting of seeds is carried out in insulating box, and seed is put into magnetized drinking water soak 2~3h, 4~5 times repeatedly, total seed soaking time 12h; The seed soaking condition: 20 ± 2 ℃ of temperature, illumination are 2000~2200lux.Through the vernalization processing selfheal percentage of seedgermination is reached more than 95%.
(3) artificial breeding: treat that 70% plant grows 2 true leaves, begin to be positioned over seeding and seedling raising in the husky bed in greenhouse, grow seedlings and select the cave dish for use, matrix is river sand: vermiculite: peat=1: 1: 1, normal water and fertilizer management.
(4) transplant planting: chose growth basically identical seedling (6 true leaves) on March 20th, 2010, root is cleaned, be put in the water planting case that fills nutrient solution with distilled water; Medium evenly passes through gas 1h with air pump every day continuously, is installed with 15 apertures of diameter 3cm on the plastic plate of medium plastic containers.Every strain plant is fixed in the aperture with the middle rubber stopper that has aperture of riving, and per two seedling water planting spacings are 10cm.
(5) nutrient solution concentration preparation: on Hogland culture fluid (Hogland and Arnon, 1950) basis, establish the plain 6.0mmolL of potassium
-1, used phosphorus element is potassium chloride (KCl) (analyzing pure>99%).Salt fractional precipitation and nutrition are deficient to be lacked in order to avoid, and every 7d changes culture fluid, and regularly adds distilled water to keep culture fluid concentration and volume.All medium pH values adopt Sartorius pH measuring instrument to measure utilization HCl and NaOH regulator solution pH value to 6~6.5.
(6) booth water planting case is cultivated: water planting plant top 5m place covering one deck light transmittance is 40%~45% black sunshade net, makes it to grow in appropriateness and shelters from heat or light in the environment.Plant is from water planting at ambient temperature on 28 days~June 20 March in 2010, and in results complete stool on June 20th, 2010, takes fruit ear, and routine is dried.
The present invention adopts water culture to produce selfheal; Under the water planting condition of suitable selfheal growth; Add the potassium in the suitable quantity of water cultivation nutrient fluid; Its individual plant fruit ear biomass significantly increases, and main active output (like Rosmarinic acid, ursolic acid, oleanolic acid, general flavone, water extraction) is significantly increased.In the large tracts of land cultivating process, can enhance productivity in a short time through the plain regulation measure of an amount of potassium, make that the selfheal growth is vigorous, the fruit ear commodity property is good, stable yield, active component output is high; Can be medical and health and processing and extract the raw medicinal material that the production of industry provides high-quality.
Embodiment
1. test method
(1) collect seed: will collect selfheal P.vulgaris L. seed, and select seed even, full, no damage by disease and insect, dehydration makes water content be lower than 12%, and sealing is preserved;
(2) presoaking and germinating: the selfheal presprouting of seeds is carried out in insulating box, and seed is put into magnetized drinking water soak 2~3h, 4~5 times repeatedly, total seed soaking time 12h; The seed soaking condition: 20 ± 2 ℃ of temperature, illumination are 2000~2200lux.Through the vernalization processing selfheal percentage of seedgermination is reached more than 95%.
(3) artificial breeding: treat that 70% plant grows 2 true leaves, begin to be positioned over seeding and seedling raising in the husky bed in greenhouse, grow seedlings and select the cave dish for use, matrix is river sand: vermiculite: peat=1: 1: 1, normal water and fertilizer management.
(4) transplant planting: treat that the selfheal seedling grows to the 2nd year, in late Febuary, 2010 to early March, choose growth basically identical seedling (6 true leaves), with distilled water root is cleaned, be put in the water planting case that fills nutrient solution; Medium evenly passes through gas 1h with air pump every day continuously, is installed with 15 cavities of diameter 3cm on the plastic plate of medium plastic containers.Every strain plant is fixed in the aperture with the middle rubber stopper that has aperture of riving, and per two seedling water planting spacings are 10cm.The plastic containers of each processing are positioned under the shade at random, and each handles 5 repetitions, and each repeats 15 seedlings.
(5) the nutrient solution gradient is provided with: on Hogland6 culture fluid (Hogland and Arnon, 1950) basis, establish the plain gradient of 4 phosphorus (unit: mmolL respectively
-1), be followed successively by: 0 (potassium deficiency K0), 1.0 (low potassium K1), 6.0 (middle potassium K2), 40 (high potassium K3).Used phosphorus element is potassium chloride (KCl) (analyzing pure>99%).Salt fractional precipitation and nutrition are deficient to be lacked in order to avoid, and every 7d changes culture fluid, and regularly adds distilled water to keep culture fluid concentration and volume.All medium pH values adopt Sartorius pH measuring instrument to measure utilization HCl and NaOH regulator solution pH value to 6~6.5.
(6) booth water planting case is cultivated: water planting plant top 5m place covering one deck light transmittance is 40%~45% black sunshade net, makes it to grow in appropriateness and shelters from heat or light in the environment.Plant is from water planting 84d at ambient temperature on 28 days~June 20 March in 2010, and in results plant on June 20th, 2010.All fresh plant after routine is dried, are taken fruit ear, weigh, carry out chemical composition behind the fine grinding powdered and detect.
(7) chemical composition detects: Rosmarinic acid, water extraction content are with reference to " Chinese pharmacopoeia (2010 editions an one) regulation item is measured down; General flavone content detects and adopts AAS, slightly changes, (document sees reference: Liao Li etc., selfheal total flavone extracting method and distribution characteristics research [J] between population thereof. CHINA JOURNAL OF CHINESE MATERIA MEDICA, 2008,33 (6): 651-653.); Ursolic acid, oleanolic acid content adopt the HPLC method, slightly changes, (document sees reference: Fang Luo etc. and high performance liquid chromatography is measured the content of 4 kinds of active components in the selfheal medicinal material simultaneously. CHINA JOURNAL OF CHINESE MATERIA MEDICA, 2010,35 (5): 616-619.).
(8) data analysis: test data is earlier with Microsoft Excel 2003 preliminary treatments, and (Oneway ANOVA LSD) analyzes to adopt SPSS 13.0 software kits then; (the g strain of active component output
-1)=active component content (%) * individual plant fruit ear biomass (g).
2. result and analysis
By shown in the table 1, under the plain water planting conditions of different potassium, it is heavily all the highest with the complete stool dry weight to handle selfheal fruit ear dry weight, stem with middle potassium, reaches 2.962gplant respectively
-1, 1.260gplant
-1, 6.127gplant
-1, and be significantly higher than other 3 groups processing; Low potassium handles the selfheal leaf dry weight and root dry weight is then maximum, reaches 1.721gplant
-1, 0.578gplant
-1, and be significantly higher than other 3 groups processing.It is thus clear that middle potassium is handled can significantly improve selfheal individual plant fruit ear dry weight.
The plain water planting condition of the different potassium of table 1 is to influence (n=10) gplant of each organ of selfheal and complete stool biomass
-1
Annotate: significance of difference branch α=0.05 level of extracting, the different expression of same row letter longitudinal differences are remarkable, down with.
Can know that by table 2 under the plain water planting conditions of different potassium, the selfheal spike length, fringe is wide and individual plant fruit ear number is handled the highlyest with middle potassium, is respectively 3.740cm, 0.994cm, 18.100, and be significantly higher than other 3 groups processing.This shows that middle potassium is handled cocoa and significantly improved fruit ear length and width, and significantly increases individual plant fruit ear number.
The plain water planting condition of the different potassium of table 2 is to the influence (n=10) of selfheal fruit ear commodity property and individual plant fruit ear number
Can be got by table 3, under the plain water planting condition of different potassium, Rosmarinic acid, ursolic acid, oleanolic acid, general flavone, water extraction output are all the highest with middle potassium processing in the selfheal fruit ear, are respectively 1.137gplant
-1, 0.308gplant
-1, 0.053gplant
-1, 15.053gplant
-1, 58.180gplant
-1, and be significantly higher than other 3 groups processing.This shows that middle potassium is handled can significantly improve main active output in the selfheal.
The plain water planting condition of the different potassium of table 3 is to influence (n=10) gplant of active component output in the selfheal fruit ear
-1
Note: the plain horizontal nutrient solution compound method of different potassium:
Each group is handled all and is added each reagent as basic nutrient solution by the listed concentration of table 4.Each group reaches concentration requirement in the table by the KCl of table 5 adding different quality respectively then.Using NaOH or HCl to regulate pH is 6.0~6.5.
Table 4: different each constituent concentration of potassium level experimental basis nutrient solution
Annotate: above each reagent is mixed with 50~500 times of highly concentrated solutions with deionized water respectively,
Directly accurately pipette dilution during each preparing nutrient solution and obtain concentration in the table.
Table 5: different potassium level test K concentration tables (KCl MW 74.5)
Annotate: after every group of test KCl demand calculated by above concentration, accurately take by weighing,
Add in the deionized water and be made into the above demand experimental concentration of respectively organizing.
Quality is every liter of nutrient solution reagent demand in the table, needs according to actual dose volume conversion.
Accompanying drawing 1 explanation: the plain water planting condition of different potassium is to the influence of selfheal growth.
Claims (2)
1. cultivation method that improves selfheal fruit ear biomass and active component output, it may further comprise the steps:
(1) collect seed: collect selfheal Prunella vulgaris Linn seed, select seed even, full, no damage by disease and insect, dehydration makes water content be lower than 12%, and sealing is preserved;
(2) presoaking and germinating: in insulating box, carry out, seed is put into magnetized drinking water soak 2~3h, 4~5 times repeatedly, total seed soaking time 12h; The seed soaking condition: temperature is 20 ± 2 ℃, and illumination is 2000~2200lux, handles making the selfheal percentage of seedgermination be higher than 95% through vernalization;
(3) artificial breeding: when 70% plant grows 2 true leaves, be positioned over and adopt cave dish seeding and seedling raising in the husky bed in greenhouse October then, matrix is river sand: vermiculite: peat=1: 1: 1, normal water and fertilizer management;
(4) transplant planting: late Febuary next year is chosen the seedling that grows 6 true leaves to early March, with distilled water root is cleaned, and is put in the water planting case that fills nutrient solution; Medium evenly passes through gas 1h with air pump every day continuously; On the plastic plate of medium plastic containers, be installed with 15 apertures of diameter 3cm, every strain plant is fixed in the aperture, and per 2 young plant water planting spacings are 10cm, and each is handled plastic containers and is positioned under the shade at random;
(5) nutrient solution concentration is set: configuration potassium cellulose content is 6.0mmolL
-1The Hogland culture fluid, the potassium element disposes with potassium chloride (KCl); Every 7d changes culture fluid, and regularly adds distilled water and keep culture fluid concentration and volume; With HCl and NaOH regulator solution pH 6~6.5;
(6) booth water planting case is cultivated: it is 40%~45% black sunshade net that 5m place, water planting plant top covers one deck light transmittance; Plant is cultivated in booth water planting case mid-March next year to mid-June, and take fruit ear during the fruit ear brownish red mid-June; Naturally dry, put aeration-drying place storage.
2. according to right 1 said a kind of ciltivating process that improves selfheal fruit ear biomass and active component output, it is characterized in that: adopt this ciltivating process, selfheal individual plant fruit ear biomass is 2.962gplant
-1Rosmarinic acid, ursolic acid, oleanolic acid, general flavone, water extraction output are respectively 1.137gplant in the fruit ear
-1, 0.308gplant
-1, 0.053gplant
-1, 15.053gplant
-1, 58.180gplant
-1
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CN105493867A (en) * | 2015-12-25 | 2016-04-20 | 南京农业大学 | Technology for continuously culturing chrysanthemum morifolium ramat and prunella vulgaris in Subei coastal areas |
CN105981556A (en) * | 2016-01-25 | 2016-10-05 | 安徽亿源药业股份有限公司 | Cultivation method of prunella vulgaris |
CN107787774A (en) * | 2017-11-03 | 2018-03-13 | 南京农业大学 | A kind of uv b radiation technology for improving selfheal fruit ear biomass and its medical material quanlity |
CN117088727A (en) * | 2023-10-18 | 2023-11-21 | 云南省农业科学院药用植物研究所 | Nutrient solution and method for improving domestication survival rate and growth rate of tsaoko tissue culture seedlings |
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Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105493867A (en) * | 2015-12-25 | 2016-04-20 | 南京农业大学 | Technology for continuously culturing chrysanthemum morifolium ramat and prunella vulgaris in Subei coastal areas |
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CN107787774A (en) * | 2017-11-03 | 2018-03-13 | 南京农业大学 | A kind of uv b radiation technology for improving selfheal fruit ear biomass and its medical material quanlity |
CN117088727A (en) * | 2023-10-18 | 2023-11-21 | 云南省农业科学院药用植物研究所 | Nutrient solution and method for improving domestication survival rate and growth rate of tsaoko tissue culture seedlings |
CN117088727B (en) * | 2023-10-18 | 2023-12-15 | 云南省农业科学院药用植物研究所 | Nutrient solution and method for improving domestication survival rate and growth rate of tsaoko tissue culture seedlings |
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