CN106912349A - A kind of method that utilization fungi promotes galuteolin accumulation in In Flower Buds of Lonicera Japonica Thunb - Google Patents
A kind of method that utilization fungi promotes galuteolin accumulation in In Flower Buds of Lonicera Japonica Thunb Download PDFInfo
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- cuttage
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- honeysuckle
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- 238000000034 method Methods 0.000 title claims abstract description 47
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- PEFNSGRTCBGNAN-QNDFHXLGSA-N luteolin 7-O-beta-D-glucoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC(O)=C2C(=O)C=C(C=3C=C(O)C(O)=CC=3)OC2=C1 PEFNSGRTCBGNAN-QNDFHXLGSA-N 0.000 title claims abstract description 22
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G17/00—Cultivation of hops, vines, fruit trees, or like trees
- A01G17/005—Cultivation methods
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- Life Sciences & Earth Sciences (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Cultivation Of Plants (AREA)
Abstract
The invention discloses a kind of method that utilization fungi promotes galuteolin accumulation in In Flower Buds of Lonicera Japonica Thunb, the method cultivates honeysuckle using cuttage seedling raising method, and carries out fungi inoculation to honeysuckle branch during cuttage, to improve the content of galuteolin in In Flower Buds of Lonicera Japonica Thunb；The fungi is the lobate layer bacterium of currant and aspergillus oryzae.The present invention is processed honeysuckle plant with fungi, makes the growth with fungi in honeysuckle growth course, and fungi is grown in the stem foot position of honeysuckle, and the growth of fungi can promote the accumulation of secondary metabolite in In Flower Buds of Lonicera Japonica Thunb, the quality of raising honeysuckle.The inventive method raw material sources are extensive, easy to operate, it is easy to implement, significant to improving quality of Flos Lonicerae in production.
Description
Technical field
The present invention relates to a kind of method for promoting secondary metabolite accumulation in In Flower Buds of Lonicera Japonica Thunb, and in particular to one kind is using true
The method that bacterium promotes galuteolin accumulation in In Flower Buds of Lonicera Japonica Thunb.
Background technology
With being filled with for " back to nature " trend, traditional natural plant is increasingly favored by people.Honeysuckle source
In caprifoliaceae plant honeysuckleLonicera japonicaThunb. the flower that dry flower or band is just opened, it is cold in nature, it is sweet, have
The effect of clearing heat and detoxicating, wind-heat dissipating, be tcm clinical practice common medicine, has used honeysuckle in about 1/3 Chinese medicinal formulae according to statistics,
And the extensive use in the Chinese patent drugs such as swap buffers, antitoxic bolus of honeysuckle flower and forsythia, oral liquid of honeysuckle flower and scutellaria, it is especially great in SARS and bird flu etc.
Great function is played in Prevention and cure of epidemic situation, has been described as " penicillin in Chinese medicine ".Demand of the current domestic and international market to honeysuckle
Amount increasingly increases, the also more and more higher of the requirement to quality of medicinal material.Secondary metabolites in medicinal plant body is typically Chinese medicine
Play the important substance basis of clinical efficacy, 2015 editions《Chinese Pharmacopoeia》Galuteolin content in middle regulation honeysuckle body is by conduct
The accumulation for weighing galuteolin in one of important indicator of traditional Chinese medicine honeysuckle quality, therefore promotion honeysuckle body of adopting an effective measure is right
It is more and more important in the quality for improving traditional Chinese medicine honeysuckle.
The lobate layer bacterium of currantPhylloporia ribis(Schumach.:Fr.) Ryvarden is derived from honeysuckle
A kind of fungi on plant, the bacterium main product parasitizes honeysuckle plant dry or exposed root, its fructification quilt always in Pingyi, shandong Province
Referred to as " honeysuckle flower moth ".The fungi is a kind of medicinal fungi, is had a long history in locality, is loaded within 2002《In Shandong Province
Medicinal material standard》, it contains the active ingredients such as triterpenes, sterols, polysaccharide and styryl pyranone, with heat-clearing solution
The effect such as poison, detumescence relieving sore-throat, hypoglycemic and antitumor.At present, the research to the lobate layer bacterium of currant is less, is mainly made
Into food or medicinal raw material, otherwise purposes has no report.
The content of the invention
It is an object of the invention to provide a kind of utilization fungi promote In Flower Buds of Lonicera Japonica Thunb in galuteolin accumulation method, the method be
The lobate layer bacterium of currant and aspergillus oryzae are inoculated with honeysuckle plant, to improve secondary metabolite --- galuteolin in In Flower Buds of Lonicera Japonica Thunb
Content, the method to improve traditional Chinese medicine honeysuckle quality it is significant.
The lobate layer bacterium of currantPhylloporia ribis(Schumach.:Fr.) Ryvarden is to be grown on honeysuckle
A kind of original wild fungi on plant, finds by research, the currant leaf grown on original wild state lower honeysuckle plant
Accumulation of the shape layer bacterium on secondary metabolites in In Flower Buds of Lonicera Japonica Thunb does not make a significant impact, and this may be with honeysuckle and the lobate layer of currant
It is relevant with the balance of ecology that bacterium long-term co-existence maintains relative material.Inventor by research, by the lobate layer bacterium of currant with
Aspergillus oryzae is cooperatively seeded on honeysuckle plant, and both bacterium co-inoculations can promote secondary metabolite in In Flower Buds of Lonicera Japonica Thunb
Accumulation, so that the content of secondary metabolites is greatly improved in In Flower Buds of Lonicera Japonica Thunb.
Concrete technical scheme of the present invention is as follows：
A kind of method that utilization fungi promotes galuteolin accumulation in In Flower Buds of Lonicera Japonica Thunb, the method is：Cultivated using cuttage seedling raising method
Honeysuckle, and fungi inoculation is carried out to honeysuckle branch during cuttage, to improve the content of galuteolin in In Flower Buds of Lonicera Japonica Thunb；Institute
Fungi is stated for the lobate layer bacterium of currant and aspergillus oryzae.
The present invention to honeysuckle branch during cuttage by way of carrying out fungi inoculation come in improving In Flower Buds of Lonicera Japonica Thunb times
The accumulation of raw metabolite galuteolin, so as to improve its content.The method of cuttage and seedling culture of the present invention is comprised the following steps：
（1）First 2 days of cuttage are poured with the carbendazim aqueous solution that mass concentration is 1/3000 to matrix and slotting machine, until base
Matter and slotting machine are wet through；
（2）Then matrix and slotting machine are exposed to the sun 2 days under the sun, matrix is put in slotting machine；
（3）Matrix and slotting machine are poured with fungi bacterium solution within 2 hours before cuttage, until matrix and slotting machine are all irrigated, it is described
Fungi bacterium solution is the mixed liquor of the lobate layer bacterium bacterium solution of currant and aspergillus oryzae bacterium solution；
（4）By honeysuckle cuttage branch sterilization, it is placed in fungi bacterium solution and soaks 10-12h, root-inducing powder is dipped after taking-up, then
Cuttage is carried out, is poured after cuttage once permeable；
（5）After cuttage, cuttage and seedling culture management is carried out to honeysuckle cuttage branch, until cuttage seeding grows to meet transplanting or go out garden wanting
Ask, then transplanted and seedling management satisfactory cuttage seeding（That is field management）, plant budding after 15-20 days
Pluck bud.
In the inventive method, honeysuckle plant is inoculated with using fungi bacterium solution, the fungi bacterium solution is that currant is lobate
The mixed liquor of layer bacterium bacterium solution and aspergillus oryzae bacterium solution.Wherein, the lobate layer bacterium bacterium solution of the currant can be with wild state lower honeysuckle
The lobate layer bacterium Phylloporia ribis (Schumach. of currant grown on plant:Fr.) Ryvarden fructifications are
Spawn incubation is obtained, it is also possible to take preserving number as the lobate layer bacterium Phylloporia ribis of the currant of CGMCC NO 1195
(Schumach.:Fr.) Ryvarden is obtained for Spawn incubation；The aspergillus oryzae bacterium solution can be obtained by Spawn incubation of aspergillus oryzae
Arrive, the aspergillus oryzae can be bought from market.
Further, the preparation method of the lobate layer bacterium bacterium solution of currant used by the present invention is：
A. by the lobate layer bacterium of currantPhylloporia ribis(Schumach.:Fr.) Ryvarden is inoculated into inclined-plane culture
On base, cultivated 5-7 days at 29 DEG C；The slant medium is made up of the component of following weight percentage：Potato 20%, agar
1.5%th, glucose 2%, potassium dihydrogen phosphate 0.2%, magnesium sulfate 0.05%, vitamin 1 × 10-3%, water surplus；
B. the lobate layer bacterium hypha body of preferable currant that will be grown after inclined-plane culture is inoculated into fluid nutrient medium, 29 DEG C,
Cultivated 5-6 days under conditions of 150rpm, obtain the lobate layer bacterium bacterium solution of currant, per inoculation 5-8 of every bottle of 1000ml fluid nutrient mediums
Hypha body；The fluid nutrient medium is made up of the component of following weight percentage：Potato 30%, glucose 2%, agar 1.4%,
Brewer's wort（10Bx）10%th, more than sucrose 1.5%, peptone 0.2%, wheat bran 4%, potassium dihydrogen phosphate 0.05%, magnesium sulfate 0.025%, water
Amount.
Further, the preparation method of the aspergillus oryzae bacterium solution used by the present invention is：
（1）Aspergillus oryzae is inoculated on slant medium, it is incubated 3-5 days at 25-30 DEG C；The slant medium is by following
The component composition of weight percentage：Potato 20%, glucose 2%, agar 1.6%, peptone 0.2%, potassium dihydrogen phosphate 0.2%, seven
Water magnesium sulfate 0.1%；
（2）Strain obtained by inclined-plane culture is inoculated into fluid nutrient medium according to the inoculum concentration of 5-10wt%, 25-30 DEG C,
Cultivated 3-5 days on the shaking table of 50-150r/min, obtain aspergillus oryzae bacterium solution；The fluid nutrient medium is by following weight percentage
Component is constituted：Rice bran 0.8%, sucrose 0.5%, glucose 1.5%, dusty yeast 0.6%, frerrous chloride 0.015%, magnesium sulfate 0.10%,
Dipotassium hydrogen phosphate 0.15%, water surplus.
In the above-mentioned fungi bacterium solution of the present invention, the volume ratio of the lobate layer bacterium bacterium solution of currant and aspergillus oryzae bacterium solution is 2-8:1, it is excellent
Elect 5 as:1.The lobate layer bacterium bacterium solution of currant and aspergillus oryzae bacterium solution are the bacterium solutions for obtaining according to the method described above, will both bacterium
Liquid mixes according to the volume ratio of regulation, you can obtain fungi bacterium solution.
In above-mentioned cottage method, step（1）In matrix used be river sand, vermiculite, fertile soil in mass ratio 2: 1: 1 mixing institute
.
In above-mentioned cottage method, the preparation method of honeysuckle cuttage branch is as follows：The selection cloudy morning 9:00 or so collection skewer
Transplant bar, the biennial 4mm of clip thick robust growth, the spray of no disease and pests harm or stiff wood are cut into the cutting of 30cm long or so, often
There is 6-8 section in individual cutting, blade of each section position each cuts 1/2 leaf, cuttage fringe bar inferior end notch wants smooth for oblique
Mouthful, 50 are bundled into 1 bundle, are squirted branch with sprayer, after after branch all moistening, are placed into foam heat-insulating chest, on
Face one layer of wet towel moisturizing of lid again, closes the lid, and waits cuttage.
In above-mentioned cottage method, step（4）Middle honeysuckle cuttage branch soaks 10- with the potassium permanganate solution of 0.06wt%
12 h, carry out sterilization.Honeysuckle cuttage branch is soaked in fungi bacterium solution after sterilization, so that as far as possible many fungies connect
Plant in cuttage branch, cuttage is carried out by root-inducing powder is dipped after immersion.The root-inducing powder is promotion branch commonly used in the art
The inorganic agent taken root, can voluntarily configure, it is also possible to commercially.
In above-mentioned cottage method, during cuttage, the angle end of cuttage branch is inserted perpendicularly into seedling bed, matrix is injected in lower end
8-12cm, cutting density is 0.5 × 0.25cm.Compacting is tracked tramping after cuttage, 1 water is irrigated.Watered in time afterwards, applied fertilizer
Deng Routine Management, 1 water was typically poured every 5 days, can take root within 7-15 days and rudiment start growth, after surviving by transplanted,
Conventional field management.
In the inventive method, after cuttage branch meets transplanting condition, the soil is porous, fertile, well-drained chiltern for selection
Transplanted in the place that loam and irrigation are convenient, have water source.More than soil 30cm is ploughed deeply after selection of land, is broken up the clods, leveling rake is thin,
Use sufficient base manure.Then the high ridge or high ridge for making 1.3m wide are transplanted.Planting-site can be utilized by barren hill, rand, ditch, the preceding room in room
The fragmentary plot such as afterwards, before typically being sprouted in early spring or rest period autumn and winter is transplanted.During transplanting, pressed on whole good planting-site
Line-spacing 150cm, spacing in the rows 120cm dig cave, and each 30-40cm of depth wide is general per hm23750~4200 caves of plant.Often spreading manuer in holes, it is miscellaneous to bury
Fertile 5kg is mixed thoroughly with subsoil, and 1 plant of strong sprout is then planted per cave, fills out fine earth compression, having peace of mind, irrigates root water, is then watered every 30 days
Once, and conventional field management is carried out, is plucked by 15-20 days after buddingging.Pluck preferably in the daily morning, do not sandwiched during harvesting
Branches and leaves impurity, preferably handles with care, and it is to be detected that the fresh flower after adopting should in time place 40-50 DEG C of low temperature drying.
The present invention is processed honeysuckle plant with fungi, makes the growth with fungi in honeysuckle growth course, fungi life
At the stem foot position of honeysuckle, the growth of fungi can promote secondary metabolite in In Flower Buds of Lonicera Japonica Thunb to length --- the product of galuteolin
It is tired, improve the quality of honeysuckle.The inventive method raw material sources are extensive, easy to operate, it is easy to implement, to improving gold and silver in production
Flower quality is significant.
Specific embodiment
The present invention is further described below by specific embodiment, the description below merely to explain the present invention,
Its content is not limited.
Embodiment 1
Honeysuckle is cultivated using following methods, takes bud：
1st, the preparation of honeysuckle cuttage branch
Early April, the selection cloudy morning 9:00 or so collection cuttage branch, chooses the thick robust growth of biennial 4mm, without disease pest
Evil without, without drying shrinkage, without broken skin damage healthy and strong spray or stiff wood, be cut into the cutting of a length of 30cm or so, have 6-8 in each cutting
Individual section, the blade of each section position each cuts 1/2 leaf, and it is angle that cuttage fringe bar inferior end notch wants smooth.By the branch of clip
Every 50 are bundled into 1 bundle, are squirted branch with sprayer, after after branch all moistening, are placed into foam heat-insulating chest, on
Face one layer of wet towel moisturizing of lid again, closes the lid, and waits cuttage.
, fungi bacterium solution preparation
2.1 prepare the lobate layer bacterium bacterium solution of currant
A. the preparation of culture medium
The preparation of slant medium：Potato is commercially bought, crust of pruning is cut into 1cm square fritters, and potato block is added
In water, 30min is boiled after 121 DEG C of sterilizings（Note the control of firepower, can suitably moisturizing）, filtrate is taken with filtered through gauze.In filtrate
Middle addition agar, glucose, potassium dihydrogen phosphate, magnesium sulfate, vitamin, are stirred continuously with glass bar, and heating for dissolving supplies water
Point, adjustment pH value to 6 loads test tube, is cooled down after sterilizing, bevel culture medium；Inclined-plane culture based formulas are（wt%）：Potato
20%th, agar 1.5%, glucose 2%, potassium dihydrogen phosphate 0.2%, magnesium sulfate 0.05%, vitamin 1 × 10-3%, water surplus.
The preparation of fluid nutrient medium：Potato is commercially bought, crust of pruning is cut into 1cm square fritters, by potato block
It is added to the water, 30min is boiled after 121 DEG C of sterilizings（Note the control of firepower, can suitably moisturizing）, filtrate is taken with filtered through gauze.
Glucose, agar, brewer's wort are added in filtrate（10Bx）, sucrose, peptone, wheat bran, potassium dihydrogen phosphate, magnesium sulfate, supply water
Point, adjustment pH value to 6, sterilizing obtains fluid nutrient medium；Liquid Culture based formulas are（wt%）：Potato 30%, glucose 2%, agar
1.4%, brewer's wort（10Bx）10%, sucrose 1.5%, peptone 0.2%, wheat bran 4%, potassium dihydrogen phosphate 0.05%, magnesium sulfate
0.025%, water surplus.
B. the preparation of the lobate layer bacterium bacterium solution of currant
Inoculation：Inoculation is operated on clean work station and carries out.When lobate with the currant grown on wild state plant of lower honeysuckle
Layer bacteriumPhylloporia ribis(Schumach.:Fr. when) Ryvarden fructifications are strain, uviol lamp is first used before inoculation
Sterilize 40min to fructification, and the portion of tissue that fructification edge is growing is inoculated in containing inclined-plane culture under aseptic conditions
In the test tube of base, cultivated 5-7 days at 29 DEG C.When the lobate layer bacterium of currant with preserving number as CGMCC NO 1195Phylloporia ribis(Schumach.:Fr. when) Ryvarden is strain, directly the strain is inoculated in containing inclined-plane training
Support in the test tube of base, cultivated 5-7 days at 29 DEG C.
Liquid Culture：The lobate layer bacterium hypha body of the preferable currant of solid culture growing state enters in taking above-mentioned slant tube
Row inoculation, is inoculated into fluid nutrient medium, notes rejecting the slant medium that agar is contained in bottom, and the diameter of hypha body is about
1-2mm, is as far as possible consistent the size of taken hypha body, and 8 hypha bodies are inoculated with per 1000ml fluid nutrient mediums, will after inoculation
Triangular flask is placed in 29 DEG C, is cultivated 6 days under conditions of rotating speed 150rpm, obtains the lobate layer bacterium bacterium solution of currant.
2.2 prepare aspergillus oryzae bacterium solution
A. the preparation of culture medium
The preparation of slant medium：Potato is commercially bought, crust of pruning is cut into 1cm square fritters, and potato block is added
In water, 30min is boiled after 121 DEG C of sterilizings（Note the control of firepower, can suitably moisturizing）, filtrate is taken with filtered through gauze.In filtrate
Middle addition glucose, agar, peptone, potassium dihydrogen phosphate, epsom salt, are stirred continuously with glass bar, and heating for dissolving is supplied
Moisture, adjustment pH value to 6 loads test tube, is cooled down after sterilizing, bevel culture medium；Inclined-plane culture based formulas are（wt%）：Soil
Beans 20%, glucose 2%, agar 1.6%, peptone 0.2%, potassium dihydrogen phosphate 0.2%, epsom salt 0.1%.
The preparation of fluid nutrient medium：Liquid Culture based formulas are（wt%）：Rice bran 0.8%, sucrose 0.5%, glucose 1.5%,
Dusty yeast 0.6%, frerrous chloride 0.015%, magnesium sulfate 0.10%, dipotassium hydrogen phosphate 0.15%, water surplus.Each component is mixed, 121
DEG C sterilizing after boil 30min（Note the control of firepower, can suitably moisturizing）Adjustment pH to 6, sterilizing obtains fluid nutrient medium.
B. the preparation of aspergillus oryzae bacterium solution
Inoculation：Inoculation is operated on clean work station and carries out.With aspergillus oryzae（It is limited purchased from Jining of Shandong Province city gold beneficial bacteria biotechnology
Company）It is strain, directly the strain is inoculated in the test tube containing slant medium, is cultivated 3-5 days at 30 DEG C.
Liquid Culture：Strain obtained by inclined-plane culture is inoculated into fluid nutrient medium according to the inoculum concentration of 8wt%, 30
DEG C, cultivate 5 days on the shaking table of 50-150r/min, obtain aspergillus oryzae bacterium solution.
2.3 prepare fungi bacterium solution
The lobate layer bacterium bacterium solution of currant obtained above and aspergillus oryzae bacterium solution are taken, by them according to 5:1 volume ratio is well mixed,
Obtain final product fungi bacterium solution.
, honeysuckle cuttage and seedling culture bed preparation
Honeysuckle cuttage educates slotting machine using 2m long, and the bottom of 1m wide, 0.25m high has the plastic containers of leaking hole, built-in culture
Matrix is river sand, vermiculite, fertile soil, and mass ratio is to mix at 2: 1: 1.First 2 days of cuttage are 1/3000 with mass concentration
The carbendazim aqueous solution is poured to matrix and slotting machine, until matrix and slotting machine are wet through.Then by matrix and slotting machine too
It is exposed to the sun under sun 2 days, matrix is put in slotting machine.Poured with fungi bacterium solution within 2 hours before cuttage, until matrix and slotting machine whole
Irrigate.
, honeysuckle branch cuttage
Honeysuckle cuttage branch is put into the potassium permanganate solution of 0.06wt%, 10-12 h are soaked, sterilization is carried out.Then
It is placed in fungi bacterium solution（The lobate layer bacterium bacterium solution of currant and aspergillus oryzae bacterium solution volume ratio are 5:1）Middle immersion 12h, dips life after taking-up
Root powder, speed is stained with 20S, then carries out cuttage.During cuttage, the angle end of cuttage branch is inserted perpendicularly into seedling bed, lower end is injected
Matrix 8-12cm, cutting density is generally 0.5 × 0.25cm.Compacting is tracked tramping after cuttage, 1 water is irrigated.Carry out in time afterwards
The Routine Management such as watered, apply fertilizer, and 1 water was typically poured every 5 days, can take root within 7-15 days and rudiment starts growth, after surviving
Transplanted.
, transplant process
After cuttage branch meets transplanting condition, selectively the soil is porous, fertile, well-drained sand loam and irrigation side for selection
Just transplanted in the place for, having water source.More than soil 30cm is ploughed deeply after selection of land, is broken up the clods, leveling rake is thin, uses sufficient base manure.So
The high ridge or high ridge for making 1.3m wide afterwards are transplanted.Planting-site can be utilized by barren hill, rand, ditch, the fragmentary plot of Around the house,
It is general sprouted in early spring before or rest period autumn and winter transplanted.During transplanting, line-spacing 150cm, strain are pressed on whole good planting-site
Cave is dug away from 120cm, each 30-40cm of depth wide is general per hm23750~4200 caves of plant.Often spread manuer in holes and mixed with subsoil into farmyard manure 5kg
It is even, 1 plant of strong sprout is then planted per cave, fine earth compression, having peace of mind are filled out, root water is irrigated, then watered once every 30 days, and carry out often
Rule field management.Band plant is long to pluck to after buddingging by 15-20 days.Pluck preferably in the daily morning, branch is not sandwiched during harvesting
Leaf impurity, preferably handles with care, and it is to be detected that the fresh flower after adopting should in time place 40-50 DEG C of low temperature drying.
Embodiment 2
Method according to embodiment 1 is cultivated to honeysuckle, gathers bud, unlike：The lobate layer bacterium bacterium solution of currant and rice
The volume ratio of aspergillus bacterium solution is 2:1.
Embodiment 3
Method according to embodiment 1 is cultivated to honeysuckle, gathers bud, unlike：The lobate layer bacterium bacterium solution of currant and rice
The volume ratio of aspergillus bacterium solution is 8:1.
Comparative example 1
Method according to embodiment 1 is cultivated to honeysuckle, gathers bud, unlike：Fungi bacterium solution is single tea
Fischer cotyledon shape layer bacterium bacterium solution.
Comparative example 2
Method according to embodiment 1 is cultivated to honeysuckle, gathers bud, unlike：Fungi bacterium solution is single rice
Aspergillus bacterium solution.
Comparative example 3
Method according to embodiment 1 is cultivated to honeysuckle, gathers bud, unlike：Fungi bacterium solution is that volume ratio is
5:The 1 lobate layer bacterium bacterium solution of currant and the mixed liquor of bacillus megaterium bacterium solution.
The preparation method of bacillus megaterium bacterium solution is：
According to the inoculum concentration of 1wt% by bacillus megaterium（Purchased from Yangzhou Haicheng Bioisystech Co., Ltd）It is inoculated into liquid
In culture medium, adjustment temperature is 35-38 DEG C, and aseptic Boiler pressure control ferments 2 days in 90-100m3/h.Liquid Culture based formulas are：
Corn flour 1.2%, dusty yeast 0.3%, beancake powder 0.4%, sodium chloride 0.05%, magnesium sulfate 0.06%, dipotassium hydrogen phosphate 0.2%, carbonic acid
Calcium 0.1%, water surplus, 121 DEG C of sterilizings, pH7.0-7.2.
Comparative example 4
Cuttage and seedling culture method according to embodiment 1 cultivates honeysuckle plant, gathers bud, unlike：Matrix, slotting machine and honeysuckle skewer
Bar of transplanting does not use fungi bacterium solution to be processed.
In order to verify influence of the cultural method of the present invention to bud its secondary metabolites content, cultivated in Pingyi, shandong Province
Experiment.Experimental technique is：Selection is suitable to transplant ground, polylith cell is classified as, per plot area 66.7m2, using random area
Group arrangement, every piece of cell cultivates honeysuckle, every kind of cultural method according to a kind of method in embodiment 1-3 and comparative example 1-4 respectively
It is repeated 4 times.Except the treatment of fungi bacterium solution is different outer, other seedling managements are kept unanimously, such as fertilising, dispenser, water, remove
Grass.
In experimentation, the galuteolin in bud is examined after the upgrowth situation, the collection bud that record honeysuckle plant
Survey, evaluate influence of the various methods to galuteolin content.
Galuteolin detection method of content is as follows：
1st, the preparation of reference substance solution：Take galuteolin reference substance appropriate, it is accurately weighed, in putting brown measuring bottle, plus 70% ethanol
Solution of every 1ml containing 40 μ g is made, is obtained final product (less than 10 DEG C preservations).
2nd, the preparation of sample solution：Bud sample grinds after 40-50 DEG C of low temperature drying is placed in after adopting, and takes required determination sample
Fine powder (cross No. 4 sieve) about 2g, accurately weighed, in putting conical flask with cover, precision adds the ml of 70% ethanol 50, weighed weight,
It is ultrasonically treated（Power 250W, frequency 35kHz）1 h, is let cool, then weighed weight, and the weight of less loss is supplied with 70% ethanol, is shaken up,
Filtration.Precision measures the ml of filtrate 10, and to doing, residue is dissolved recycling design with 70% ethanol, is transferred in 5mL volumetric flasks, plus 70%
Ethanol is obtained final product to scale.Use organic film（0.2 μm of aperture）It is stand-by in filtering to liquid phase bottle.
3rd, content detection：Detected using liquid chromatography, chromatographic column is C18（4.60 mm × 250 mm, 1.7 μ
m）, it is Mobile phase B with 0.5% glacial acetic acid solution with acetonitrile as mobile phase A, the regulation according to the form below carries out gradient elution；Detection
Wavelength is 350nm.Number of theoretical plate is calculated by galuteolin peak and should be not less than 20000.Detection wavelength is 327nm, and sample size is
20uL, column temperature is 40 DEG C.
Specifically detection method is：Reference substance solution is made into different concentration, obtained with concentration by liquid chromatographic detection and
Peak area is the standard curve of transverse and longitudinal coordinate.Each sample solution is carried out into liquid chromatographic detection using same method, according to peak
Galuteolin content in areal calculation each sample.
Experimentation finds, when carrying out honeysuckle cultivation using the method for embodiment 1-3 and comparative example 2,3, the life of honeysuckle plant
Speed long is fast compared with comparative example 1 and 4, and plant buddings also earlier than comparative example 1 and 4, plant height mean height about 1.6cm or so.And
The honeysuckle plant incidence of insect disease of embodiment 1-3 and comparative example 2 and 3 is few.Bud to gathering carries out galuteolin detection, ties
It is really as shown in table 1 below, as can be seen from the table, in the bud that the method for embodiment 1-3 is obtained galuteolin content apparently higher than
The method of each comparative example, and the lobate layer bacterium bacterium solution of alone currant or aspergillus oryzae bacterium solution or other bacterium of use and currant leaf
The effect that shape layer bacterium bacterium solution is used cooperatively is not obvious, and accumulation of the fungi bacterium solution of the present invention to In Flower Buds of Lonicera Japonica Thunb secondary metabolite has
Good facilitation.
Claims (8)
1. a kind of method that utilization fungi promotes galuteolin accumulation in In Flower Buds of Lonicera Japonica Thunb, honeysuckle is cultivated using cuttage seedling raising method, its
It is characterised by：Fungi inoculation is carried out to honeysuckle branch during cuttage, to improve the content of galuteolin in In Flower Buds of Lonicera Japonica Thunb；Institute
Fungi is stated for the lobate layer bacterium of currant and aspergillus oryzae.
2. method according to claim 1, it is characterized in that：Comprise the following steps：
（1）Matrix and slotting machine are poured with the carbendazim aqueous solution that mass concentration is 1/3000 within 2 days before cuttage, until matrix
Wet through with slotting machine；
（2）Then matrix and slotting machine are exposed to the sun 2 days under the sun, matrix is put in slotting machine；
（3）Matrix and slotting machine are poured with fungi bacterium solution within 2 hours before cuttage, until matrix and slotting machine are all irrigated, it is described
Fungi bacterium solution is the mixed liquor of the lobate layer bacterium bacterium solution of currant and aspergillus oryzae bacterium solution；
（4）By honeysuckle cuttage branch sterilization, it is placed in fungi bacterium solution and soaks 10-12h, root-inducing powder is dipped after taking-up, then
Cuttage is carried out, is poured after cuttage once permeable；
（5）After cuttage, cuttage and seedling culture management is carried out to honeysuckle cuttage branch, until cuttage seeding grows to meet transplanting or go out garden wanting
Ask, then transplanted and seedling management satisfactory cuttage seeding, plant plucks bud after buddingging by 15-20 days.
3. method according to claim 2, it is characterized in that：Fungi bacterium solution is the lobate layer bacterium bacterium solution of currant and aspergillus oryzae
The mixed liquor of liquid；
The preparation method of the lobate layer bacterium bacterium solution of currant is：
A. by the lobate layer bacterium of currantPhylloporia ribis(Schumach.:Fr.) Ryvarden is inoculated into inclined-plane culture
On base, cultivated 5-7 days at 29 DEG C；The slant medium is made up of the component of following weight percentage：Potato 20%, agar
1.5%th, glucose 2%, potassium dihydrogen phosphate 0.2%, magnesium sulfate 0.05%, vitamin 1 × 10-3%, water surplus；
B. the lobate layer bacterium hypha body of preferable currant that will be grown after inclined-plane culture is inoculated into fluid nutrient medium, 29 DEG C,
Cultivated 5-6 days under conditions of 150rpm, obtain the lobate layer bacterium bacterium solution of currant, 7-8 mycelia is inoculated with per 1000ml fluid nutrient mediums
Group；The fluid nutrient medium is made up of the component of following weight percentage：Potato 30%, glucose 2%, agar 1.4%, malt
Juice（10Bx）10%th, sucrose 1.5%, peptone 0.2%, wheat bran 4%, potassium dihydrogen phosphate 0.05%, magnesium sulfate 0.025%, water surplus；
The preparation method of aspergillus oryzae bacterium solution is：
（1）Aspergillus oryzae is inoculated on slant medium, it is incubated 2-3 days at 25-30 DEG C；The slant medium is by following
The component composition of weight percentage：Potato 20%, glucose 2%, agar 1.6%, peptone 0.2%, potassium dihydrogen phosphate 0.2%, seven
Water magnesium sulfate 0.1%；
（2）Strain obtained by inclined-plane culture is inoculated into fluid nutrient medium according to the inoculum concentration of 5-10wt%, 25-30 DEG C,
Cultivated 6-7 days on the shaking table of 150-200r/min, obtain aspergillus oryzae bacterium solution；The fluid nutrient medium is by following weight percentage
Component is constituted：Rice bran 0.8%, sucrose 0.5%, glucose 1.5%, dusty yeast 0.6%, frerrous chloride 0.015%, magnesium sulfate 0.10%,
Dipotassium hydrogen phosphate 0.15%, water surplus.
4. according to the method in claim 2 or 3, it is characterized in that：The lobate layer bacterium bacterium solution of currant is with wild state
The lobate layer bacterium of currant grown on honeysuckle plantPhylloporia ribis(Schumach.:Fr.) Ryvarden are real
Body is strain, or is the lobate layer bacterium of the currant of CGMCC NO 1195 with preserving numberPhylloporia ribis
(Schumach.:Fr.) Ryvarden is strain；The aspergillus oryzae bacterium solution is with aspergillus oryzae as strain.
5. the method according to claim 2,3 or 4, it is characterized in that：In fungi bacterium solution, the lobate layer bacterium bacterium solution of currant and rice
The volume ratio of aspergillus bacterium solution is 2-8:1, preferably 5:1.
6. method according to claim 2, it is characterized in that：Step（1）In, the matrix is river sand, vermiculite, fertile soil are pressed
The mixing gained of mass ratio 2: 1: 1；Step（4）In, honeysuckle cuttage branch soaks 10-12 with the potassium permanganate solution of 0.06wt%
H, carries out sterilization.
7. method according to claim 2, it is characterized in that：The preparation method of honeysuckle cuttage branch is：The selection cloudy morning
9:00 or so collection cuttage branch, the biennial 4mm of clip thick robust growth, the spray of no disease and pests harm or stiff wood are cut into length
The cutting of 30cm or so, 6-8 section in each cutting, the blade of each section position each cuts 1/2 leaf, and cuttage fringe bar lower end is cut
It is angle that mouth is smooth, and 50 are bundled into 1 bundle, are squirted branch with sprayer, after after branch all moistening, is placed into foam heat-insulating
In chest, one layer of wet towel moisturizing of lid again, closes the lid above, waits cuttage.
8. the method according to claim 2 or 7, it is characterized in that：During cuttage, the angle end of cuttage branch is inserted perpendicularly into and is educated
In seedbed, matrix 8-12cm is injected in lower end, and cutting density is 0.5cm × 0.25cm.
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|---|---|---|---|---|
| CN108207476A (en) * | 2018-03-02 | 2018-06-29 | 芜湖欧标农业发展有限公司 | A kind of cuttage implantation methods of carpinus turczaninowii |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103749126A (en) * | 2014-01-09 | 2014-04-30 | 浙江省林业科学研究院 | Sustainable three-nerved spicebush cultivation method |
| JP5552649B2 (en) * | 2009-03-16 | 2014-07-16 | 独立行政法人農業生物資源研究所 | Method for preventing or suppressing microbial infection of plant and microbial infection resistant plant |
| CN105660098A (en) * | 2016-01-08 | 2016-06-15 | 华南师范大学 | Method for reducing honeysuckle cadmium accumulation and increasing yield of honeysuckle by means of glomus intraradices |
| CN106358821A (en) * | 2016-08-26 | 2017-02-01 | 滁州菲扬农业科技有限公司 | Cuttage treatment method in cuttage seedling culturing process of honeysuckles |
-
2017
- 2017-03-01 CN CN201710116171.4A patent/CN106912349B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP5552649B2 (en) * | 2009-03-16 | 2014-07-16 | 独立行政法人農業生物資源研究所 | Method for preventing or suppressing microbial infection of plant and microbial infection resistant plant |
| CN103749126A (en) * | 2014-01-09 | 2014-04-30 | 浙江省林业科学研究院 | Sustainable three-nerved spicebush cultivation method |
| CN105660098A (en) * | 2016-01-08 | 2016-06-15 | 华南师范大学 | Method for reducing honeysuckle cadmium accumulation and increasing yield of honeysuckle by means of glomus intraradices |
| CN106358821A (en) * | 2016-08-26 | 2017-02-01 | 滁州菲扬农业科技有限公司 | Cuttage treatment method in cuttage seedling culturing process of honeysuckles |
Non-Patent Citations (2)
| Title |
|---|
| 刘锦春 马晔 陶建平 高凯敏 梁千慧: "《丛枝菌根真菌对西南岩溶地区干旱及干湿交替下金银花根系生长的影响》", 《北京林业大学学报》 * |
| 程玉鹏，李天聪，林进华，陈琦，马爱萍，李弘琨: "《内生真菌与药用植物宿主间的交流》", 《中医药信息》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108207476A (en) * | 2018-03-02 | 2018-06-29 | 芜湖欧标农业发展有限公司 | A kind of cuttage implantation methods of carpinus turczaninowii |
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