CN103202318B - Accumulation promoter of atractylis lancea naphtha and preparation method thereof - Google Patents
Accumulation promoter of atractylis lancea naphtha and preparation method thereof Download PDFInfo
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- CN103202318B CN103202318B CN201310119807.2A CN201310119807A CN103202318B CN 103202318 B CN103202318 B CN 103202318B CN 201310119807 A CN201310119807 A CN 201310119807A CN 103202318 B CN103202318 B CN 103202318B
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- elicitor
- absidia
- atractylis lancea
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Abstract
The invention relates to a problem of production of naphtha of a medicinal plant. An accumulation promoter with a real ingredient of atractylis lancea is prepared by a fungal elicitor and an additive, so that the accumulation promoter is more conveniently acted on the atractylis lancea in other areas, so as to improve the yield of atractylis lancea naphtha in other areas, so that the atractylis lancea has the same degree of naphtha accumulation effect as the atractylis lancea produced in Maoshan, Jiangsu Province; and the atractylis lancea has the real characteristics of the atractylis lancea produced in Jiangsu even if being in difference places.
Description
Technical field
The invention belongs to agriculture using microbe technical field, specifically by thick elicitor mould for Absidia and constituent of chitosan and a certain amount of salicylic acid and CaCl
2make cocktail spray and commercialization, be applied to market.
Background technology
1. Atractylis lancea
(1) call: Mao Shu, RHIZOMA ATRACTYLODIS IANCEAE, fornicate fasicle.
(2) originate: be the rhizome of feverfew Atractylis lancea Atractylodes lancea (Thunb.) DC..
(3) characteristic: property acrid, bitter, warm, return spleen, stomach, Liver Channel.The nice and cool comparatively arid climate of happiness, is afraid of hot and humid.
(4) chemical composition: containing volatile oil, main containing Atisine chloride Atractydin (atractylodin), β-cineol (β-eudesmol), hinesol (hinesol), hydroxyl atractylone (hydroxy-atractylon) etc. in oil.
(5) function cures mainly: have drying damp and strengthening spleen, expelling wind and clearing away cold, function of improving eyesight.Cure mainly abdominal fullness and distention, have loose bowels, oedema, arthralgia due to wind-dampness, anemofrigid cold, incoordination between the spleen and the stomach, have loose bowels, syndrome of dampness-heat, arthritis with fixed pain caused by dampness and day disease, heat temperature, dysentery, time row diseases caused by external factors, the inferior disease of yellowish-white band.Fire excess from yin deficiency, haematemesis, bleeding from five sense organs or subcutaneous tissue, deficiency of vital energy hidrosis person avoid use.
(6) market status: Atractylis lancea is mainly distributed in the provinces such as Jiangsu, Hubei and Henan, and Region in Maoshan, Jiangsu Province is the producing region of Atractylis lancea genunie medicinal materials.In recent years, along with the demand of international market to rhizoma atractylodis increases day by day, being ransacked of property of Atractylis lancea is excavated, and in addition to the artificial destruction of Atractylis lancea ecotope, causes the exhaustion of Atractylis lancea resource, makes it endangered.Still there is a lot of problem in Atractylis lancea development of resources: the germination rate of Atractylis lancea seed is low, and can not provide stable provenance for implant mass, artificial cultivation is difficult, Atractylis lancea poor growth, and Atractylis lancea resistance is poor, and happiness is shady and cool, thermo-labile, intolerant to waterlogging.
2. fungal elicitor
From fungal cell, it is the material that a class can cause plant cell dynamic accumulation secondary metabolites activity.As fungal spore, mycelium, homogenate, fungal cell wall composition, fungal cultures filtrate etc.Fungal elicitor is the structure of cell surface of fungi or secretion mainly, and in chemical constitution, it mainly comprises: polysaccharide, polypeptide, glycoprotein and unsaturated fatty acid.
Fungal elicitor effectively can improve percentage of seedgermination, accelerates medicinal material growth, improves resistance, promotes that active principle accumulation maintains medical material quanlity.Plant through fungal elicitor effect often has the advantages such as growth quick, degeneration-resistant border, disease resistance, anti-animal injury, have more some fungal elicitor of competition for existence power to there being certain capacity of decomposition from poisonous substance matter than the plant without effect, therefore utilize it can solve or alleviate Auto toxicity.
Still not thorough to the understanding of Fungal Elicitor mechanism of action at present, main points of view is identified by the acceptor on plant cell membrane as a kind of outer signals by elicitor, and combine with it, thus to cause on cell membrane and series reaction in cell, the enzymatic synthesis relevant with protective plant protecting agent or activity are changed, thus cause gene expression in plants to change, finally cause synthesis and the accumulation of protective plant protecting agent.Elicitor causes on film as outer signals function cells plasma membrane and changes, to causing gene promoter in born of the same parents, the process of enzyme activity change is a cascade process, elicitor signal is passed to after in cell, is regulated the synthesis of secondary metabolite by the synthesis controlling key enzyme in secondary metabolism approach.
3. shitosan
Shitosan, poly-β mono-Isosorbide-5-Nitrae aminoglucose is a kind of cell wall polysaccharides.There is unique physico chemical property and biological function, demonstrate wide application prospect at medicine, food, functional material, cosmetics, agricultural, environmental protection, household chemicals and biological technical field, can be used to produce Biodegradable films, fiber stickiness controlling agent, binder, paper strengthening agent and the flocculant of disposing of sewage, intercalating agent etc.Shitosan can be processed through extraction by crustacean (as crab, shrimp) shell, has multiple shitosan production marketing in the market.
4. additive
Salicylic acid (Salicy lic aci d, SA) is a kind of simple aldehydes matter be extensively present in higher plant, and it synthesizes in plant corpus, and content is very low, participates in multiple physiological and biochemical procedure in regulating plant body.And increasing research shows, SA is also the degeneration-resistant signaling molecule that Plant defense responses and inducing plant react abiotic stress.Its typical physiological action is (PR s) gene expression of pathogenesis-related proteins in inducing plant body and produces system resume (SAR).In recent years, the report that SA studies in Genes For Plant Tolerance adverse circumstance is also more, as improved the salt-resistance of plant, cold resistance, heat resistance and reducing the injury etc. of heavy metal cell membrane.
Ca
2+the structure of plant cell and physiological function are played an important role, the stability of cell wall, cell membrane and embrane-associated protein can be maintained, the adjustment process participating in born of the same parents' homeostasis (homeostasis) and grow, be called as total attemperator of plant metabolism, its importance is mainly reflected in Ca
2+combine with the CAM (calmodulin) as intracellular messenger, many physiological metabolism processes of regulating plant body, especially under environment-stress, Ca
2+participate in the impression of stress signal, transmission, response and expression with calmodulin (CAM), improve the resistance of plant.
5. idiomaticity feature
Atractylis lancea suitable growth in Hills, with the hillside fields of half cloudy half light and deep, the well-drained sandy soil of soil layer best.Tame suitable area is carried out in Jiangsu and hilliness area, surrounding area thereof.Our this research emphasis is the rhizoma atractylodis being acted on the growth of non-Jiangsu by the endophyte exciton of commercialization, and make its volatile oil accumulation effect reach best, in volatile oil, atractylone, Atisine chloride Atractydin significantly improve,
-eudesmol content significantly reduces.
Summary of the invention
The object of the invention is to prepare a kind of can improve the accumulation of Atractylis lancea idiomaticity volatile oil component promoter, thus the Atractylis lancea of area, Shi Fei Jiangsu Province plantation has its idiomaticity composition equally, the accumulation of the Atractylis lancea volatile oil in area, Jiangsu Province is improved further, in volatile oil, atractylone, Atisine chloride Atractydin significantly improve
-eudesmol content significantly reduces.And then improve economic benefit and the market value of Atractylis lancea.
The present invention is by thick elicitor mould for Absidia and constituent of chitosan and a certain amount of salicylic acid and CaCl
2make cocktail spray, act on root or the blade of Atractylis lancea.The method of operating that the present invention relates to is as follows:
A kind of Atractylis lancea volatile oil accumulation promoter of the present invention, its formula is: have the mould thick elicitor 15mg of Absidia, shitosan 15mg, salicylic acid (SA) 100mg and CaCl in every premium on currency
25g; The mould thick elicitor of described Absidia is obtained by following method: get appropriate mycelium, mycelium is after suction filtration, 3 times are repeatedly rinsed with distilled water, carry out ultrasonic disruption 30min under adding the physiological saline room temperature of 10 times of volumes, then the centrifugal 10min of 3000r/min, collect supernatant; Be elicitor crude extract after 121 DEG C of high-temperature sterilization 20min, this crude extract of freeze-drying obtains thick elicitor, and described Absidia is mould is the mould CGMCC3.3538 of Absidia.
The preparation method of above-mentioned Atractylis lancea volatile oil accumulation promoter, comprises the following steps:
(1) thick elicitor is extracted
Get the mould CGMCC3.3538 mycelium of Absidia, mycelium, after suction filtration, rinses 3 times repeatedly with distilled water, carries out ultrasonic disruption 30min under adding the physiological saline room temperature of 10 times of volumes, then the centrifugal 10min of 3000r/min, collects supernatant; Be elicitor crude extract after 121 DEG C of high-temperature sterilization 20min, this crude extract of freeze-drying obtains thick elicitor;
(2) composite
By the thick elicitor of upper step gained and shitosan in mass ratio 1:1 mix, then mixture is pressed 30mg/L and mixes with water, then add 100mg/L salicylic acid and 5g/L CaCl
2make mixing spray agent.
Embodiment
Term used in the present invention, unless otherwise specified, generally has the implication that those of ordinary skill in the art understand usually.
Below in conjunction with specific embodiment, and in further detail the present invention is described according to data.These should be understood
Embodiment just in order to demonstrate the invention, but not limits the scope of the invention in any form.
Below in an example, the various process do not described in detail and method are conventional methods as known in the art.The source of agents useful for same, trade name and be necessary to list its constituent person, all indicate when occurring first, identical reagent used if no special instructions thereafter, and all with first this content indicated is identical.
Absidia described in the present invention mould (Absidia coerulea) is from Chinese General Microbiological Culture preservation administrative center
(CGMCC) obtain, ask for numbering 3.3538.
Embodiment 1:
One, the preparation process of product
Step one: cultivate Absidia mould
(1) cultural method on inclined-plane:
Appropriate absidia orchidi kind is inoculated in potato culture (murphy juice 20g, glucose 2g, agar 2g, water 100ml, pH nature), is placed in the insulating box of 30 degrees Celsius, cultivates 3 days.
(2) cultural method of liquid seeds: get liquid seed culture medium (the bottled 250ml water of 500ml triangle, yeast extract 1g, peptone 1g that appropriate (1) is inoculated in, glucose 2g, pH nature), be placed on constant-temperature table, rotating speed 180r/min, in 30 degree of cultivations 4 days.
(3) liquid fermentation culturing method: after 10% inoculum concentration inoculation, 30 degree of shaking speed 200r/min, cultivate 4 days.Step 2: extract thick elicitor
Get appropriate mycelium, mycelium, after suction filtration, rinses several times repeatedly with distilled water, carries out ultrasonic disruption 30min under adding the physiological saline room temperature of 10 times of volumes, then the centrifugal 10min of 3000r/min, collects supernatant.Elicitor crude extract is after 121 DEG C of high-temperature sterilization 20min.The thick elicitor extract of freeze-drying.
Step 3: thick elicitor dry product mixes in 1:1 ratio with commercially available shitosan.
Step 4: make mixing spray agent
The mixture of gained in step 3 is pressed 30mg/L mix with water, then add 100mg/L SA and 5g/L CaCl
2, make mixing spray agent, be loaded in sprayer container, every mu of amount of pressing 5L applies.
Two, spray method
1. the concentration of spray agent: spray agent presses 1:50 volume and water dilutes.
2. the consumption of spray agent: about 4 kilograms, every mu of ground.
3. spray time: when the morning 5 ~ 7 or afternoon 6 ~ 8 time (after rain, spray effect can be better).
Three, spray test effect
Above-mentioned mixing spray agent was sprayed at Atractylis lancea blade face after 14 days, and detect idiomaticity composition transfer, result is as table 1 and table 2:
The different Atractylis lancea of table 1. sprays the volatile oil compositions situation of change (unit μ g/g) after promoter
Volatile oil compositions after the different Atractylis lancea of table 2. sprays promoter accounts for the volatile oil total amount percentage table of comparisons (%)
Find out from above-mentioned two tables, after applying promoter, Atractylis lancea atractylone and the Atisine chloride Atractydin content in source, the Maoshan Mountain are higher, sagittol content significantly reduces, and the Atractylis lancea in source, Shaanxi, atractylone and Atisine chloride Atractydin are close to Rhizoma Atractylodis untreated fish group level, and sagittol content is also reduced to the untreated level of Maoshan Mountain Atractylis lancea, and explanation can reach authentic medicinal herbs standard.
Claims (2)
1. an Atractylis lancea volatile oil accumulation promoter, is characterized in that the formula of this promoter is: have the mould thick elicitor 15mg of Absidia, shitosan 15mg, salicylic acid 100mg and CaCl in every premium on currency
25g;
The mould thick elicitor of described Absidia is obtained by following method:
Step one: cultivate Absidia mould
(1) cultural method on inclined-plane:
Appropriate absidia orchidi kind is inoculated in potato culture, is placed in the insulating box of 30 DEG C, cultivate 3 days;
(2) cultural method of liquid seeds: get appropriate (1) and be inoculated in liquid seed culture medium, be placed on constant-temperature table, rotating speed 180r/min, cultivate 4 days in 30 DEG C; Described liquid seed culture medium is: 250ml water, yeast extract 1g, peptone 1g, glucose 2g, pH nature;
(3) liquid fermentation culturing method: after 10% inoculum concentration inoculation, 30 DEG C of shaking speed 200r/min, cultivate 4 days; Step 2: get appropriate mycelium, mycelium, after suction filtration, rinses 3 times repeatedly with distilled water, carries out ultrasonic disruption 30min under adding the physiological saline room temperature of 10 times of volumes, then the centrifugal 10min of 3000r/min, collects supernatant; Be elicitor crude extract after 121 DEG C of high-temperature sterilization 20min, this crude extract of freeze-drying obtains thick elicitor, and described Absidia is mould is the mould CGMCC3.3538 of Absidia.
2. prepare the method for Atractylis lancea volatile oil accumulation promoter as claimed in claim 1, it is characterized in that comprising the following steps:
(1) thick elicitor is extracted
Get the mould CGMCC3.3538 mycelium of Absidia, mycelium, after suction filtration, rinses 3 times repeatedly with distilled water, carries out ultrasonic disruption 30min under adding the physiological saline room temperature of 10 times of volumes, then the centrifugal 10min of 3000r/min, collects supernatant; Be elicitor crude extract after 121 DEG C of high-temperature sterilization 20min, this crude extract of freeze-drying obtains thick elicitor;
(2) composite
By the thick elicitor of upper step gained and shitosan in mass ratio 1:1 mix, then mixture is pressed 30mg/L and mixes with water, then add 100mg/L salicylic acid and 5g/L CaCl
2make mixing spray agent, spray by the amount of 5L/ every mu.
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| CN201310119807.2A CN103202318B (en) | 2013-04-08 | 2013-04-08 | Accumulation promoter of atractylis lancea naphtha and preparation method thereof |
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| CN115820430B (en) * | 2022-09-26 | 2024-04-26 | 浙江中医药大学 | Endophytic fungus of bighead atractylodes rhizome and application of endophytic fungus in bighead atractylodes rhizome suspension cell culture |
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| JP3386489B2 (en) * | 1992-05-29 | 2003-03-17 | 大日本製薬株式会社 | Phytoalexin inducer |
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