CN107873516A - The breeding method of yncaria stem with hooks - Google Patents
The breeding method of yncaria stem with hooks Download PDFInfo
- Publication number
- CN107873516A CN107873516A CN201711122593.9A CN201711122593A CN107873516A CN 107873516 A CN107873516 A CN 107873516A CN 201711122593 A CN201711122593 A CN 201711122593A CN 107873516 A CN107873516 A CN 107873516A
- Authority
- CN
- China
- Prior art keywords
- hooks
- seedling
- yncaria stem
- culture
- water
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a kind of breeding method of yncaria stem with hooks, comprise the following steps：Step 1: yncaria stem with hooks seed is cultivated into 20~25d in seed culture medium, aseptic seedlings plumular axis is obtained；Step 2: the plumular axis explant without cotyledonary node that length is 0.4~0.8cm is cut from aseptic seedlings plumular axis, lie in the first callus tissue culture base and carry out light culture, after 20~25d, it is transferred in the second callus tissue culture base and carries out light culture, obtain fresh callus；Differentiation culture is carried out Step 3: fresh callus is transferred in differential medium, obtains clump bud；Strong seedling culture is carried out Step 4: clump bud is transferred in strong seedling culture base, when individual plant seedling length is 3~4cm, is cut off in basal part of stem, is transferred in root media and carries out culture of rootage, grow 1.5~2.0cm root, obtain yncaria stem with hooks seedling.Yncaria stem with hooks that the present invention obtains is pure natural, yield is high, pharmaceutical ingredient content is high.
Description
Technical field
The present invention relates to field of plant tissue culture technique, espespecially a kind of breeding method of yncaria stem with hooks.
Background technology
Yncaria stem with hooks is one of conventional Chinese medicine, first recorded in beam for Tao Hongjing《Mingyi Bielu》, medicinal history is long.Yncaria stem with hooks
Nature and flavor are sweet, cool, have the effect of The flat liver of heat-clearing, dispelling wind and relieving convulsion, and its main chemical compositions is rhynchophyllin and isorhynchophylline.Clinically
It is widely used in the diseases such as headache, hypertension, twitch, neurasthenia.Yncaria stem with hooks is yncaria stem with hooks piece, ZHENGTIAN WAN, Baicalin, arteries and veins
The primary raw material medicine of multiple famous herbal species such as Junan and the common medicine of Chinese medicine preparation.It is bad work and rest custom cause hypertension,
High fat of blood and hyperglycemia population quantity are continuously increased, and people increasingly increase the demand of natural plant.According to incompletely statistics, only
I.e. up to more than 10000 tons of the annual demand of yncaria stem with hooks, wherein exporting to the country such as Japan, South Korea up to more than 4000 tons.Yncaria stem with hooks grows
When have a higher requirement for the water content and ambient humidity of growing location, water content is superfluous or ambient humidity too conference causes it
Root or stem addle, or even dead.And the mode coverage of rotary spray is small, be not suitable for the shrub watering to yncaria stem with hooks class；
Drop tank then needs to lay complicated multiple-way duct on the ground, takes the growing location of yncaria stem with hooks, and is difficult to maintain yncaria stem with hooks growth needs
Humidity；Using traditional watering mode poured into from ground with water pipe, water consumption is big, irrigation amount is uneven, time-consuming, is easily caused
Part rhynchoin water suction is excessive and the root that arrives is addled, and such a situation would generally apply the medicament of preventing and treating root rot, obtained yncaria stem with hooks
Not pure natural, green yncaria stem with hooks, being used as medicine for yncaria stem with hooks increases risk.In fact, existing pass through plant division, cuttage, seed
It is relatively low etc. mode and with reference to traditional watering mode cultivate yield and the medicinal ingredient of the yncaria stem with hooks of acquisition, can not meet to be incremented by
Demand.
Therefore it provides a kind of breeding method of yncaria stem with hooks pure natural, yield is high, pharmaceutical ingredient content is high is very necessary.
The content of the invention
It is an object of the invention to solve at least the above, and provide the advantages of at least will be described later.
It is a still further object of the present invention to provide a kind of breeding method of yncaria stem with hooks, it passes through animal nutrition and chemistry side
Method, multiploid induction is carried out to yncaria stem with hooks, establish yncaria stem with hooks polyploiding system, inductivity is high, and combines distinctive watering mode
The yncaria stem with hooks plant of cultivation, the increase of its pharmaceutical ingredient content, yield are more.
In order to realize according to object of the present invention and further advantage, there is provided a kind of breeding method of yncaria stem with hooks, including
Following steps：
Step 1: 20~25d of culture will be transferred in seed culture medium after yncaria stem with hooks seed successively soaking, sterilization, obtain
Aseptic seedlings plumular axis；
Step 2: the plumular axis explant without cotyledonary node that length is 0.4~0.8cm is cut from aseptic seedlings plumular axis,
Plumular axis explant is lain in the first callus tissue culture base and carries out light culture, after 20~25d, the both ends of plumular axis explant are swollen
It is big to form plumular axis callus, plumular axis callus is then transferred to progress light culture 7 in the second callus tissue culture base
~20d, obtain fresh callus；
Differentiation 20~25d of culture is carried out Step 3: fresh callus is transferred in differential medium, obtains clump bud；
Strong seedling culture is carried out Step 4: clump bud is transferred in strong seedling culture base, when individual plant seedling length is 3~4cm,
Cut off in basal part of stem, be transferred in root media and carry out culture of rootage, after 20~25d, grow 1.5~2.0cm root, obtain
To yncaria stem with hooks seedling.
Preferably, the seed culture medium is the MS culture mediums without hormone；
The first callus tissue culture base includes MS, 1.0~1.5mg/L TDZ, 0.03~0.05mg/L NAA；
The second callus tissue culture base include MS, 1.0~1.5mg/L TDZ, 0.03~0.05mg/L NAA,
80~200mg/L colchicine；
The differential medium includes MS, 0.1~0.4% activated carbon；
The strong seedling culture base includes MS, 1.0~1.5mg/L 6-BA, 0.1~0.2mg/L NAA；
The root media includes 1/2MS, 0.5~0.8mg/L IBA, 0.2~0.5mg/L NAA, 0.2% work
Property charcoal.
Preferably, the specific method soaked in step 1 is：The yncaria stem with hooks seed chosen is mounted in beanbag, is placed in
Rotating speed is to soak 4h on 200rpm shaking table；The specific method of sterilization is：Soaked yncaria stem with hooks seed is placed in superclean bench
On, with 10% 8~10min of hypochlorite disinfectant, then with sterile water wash 4~6 times.
Preferably, also 6-BA, the 0.1mg/L including 2.0mg/L of differential medium described in step 4 NAA.
Preferably, the cultivation temperature of each step is 20~25 DEG C.
Preferably, in addition to：Step 5: will be cultivated in yncaria stem with hooks seedling replanting to seedling shed, it is specially：
A, the soil in seedling shed is toasted, then soil deeptillage until soil it is fine and soft, without grume, be sprinkled into animal manure
Just, and humus soil is added, regulation pH is 6.5~8.0, and then laying depth is 5cm sand soil, and is 2m's according to width
Specification sets two row's furrow, and a width of 20cm gutter is provided between two row's furrow；
B, it is 23 DEG C to control seedling temperature of shed, and the land area that 20 × 20cm is taken according to every plant of yncaria stem with hooks seedling is moved
Plant, after the completion of transplanting, sprinkling root water, the root water includes vitamin C, the rape of 4~6 parts by weight of 7~10 parts by weight
Plain lactone, the heteroauxin of 3~6 parts by weight, the gibberellin of 2~7 parts by weight, the sodium selenite of 5~9 parts by weight, 4~7 weight
The sodium molybdate, 3~5 parts of compound amino acids, 0.4~0.9 part of garlic selenium polysaccharide, the egg vinegar liquid of 6~11 parts by weight, 100 weights of part
The water of amount part mixes；
C, afterwards, every watering in 4 days once, every ventilation in 2 days once, control in seedling shed relative humidity for 80%~
85%.
Preferably, watering system, the watering system are used in step b and step c, when sprinkling root water and watering
Be arranged in the seedling shed, the watering system include two water-supply-pipes, N number of slideway and 2N pour water assembly, described two
The horizontal top in the seedling shed of water-supply-pipe horizontal parallel, and the width of the furrow is each perpendicular to, on every water-supply-pipe
Equidistantly be provided with N number of apopore along pipe range direction, N number of equidistant horizontal parallel of slideway be erected at two water-supply-pipes it
Between, each slideway is vertical with two water-supply-pipes, and two ends of a slideway correspond to two water-supply-pipes each
An apopore, set on a slideway two to pour water assembly, one for pouring water assembly and including being slidably connected with slideway
Sliding block, be fixedly connected with a slide block an electric expansion bar and a watering tube, one end of watering tube and slideway one of them
Apopore sealing connection, the other end corresponding to end are to be connected with the free end of electric expansion bar with strip spray end
Shower nozzle；
During watering, two sliding blocks in a slideway drive respective watering tube to slide into the two of wherein row's furrow respectively
Side, electric expansion bar elongation make shower nozzle close to yncaria stem with hooks plant, and the distance on shower nozzle and ground in seedling shed is 15~20cm, any two
Spacing between adjacent slideway is equal to the length at the strip spray end of the shower nozzle.
The present invention can at least reach following beneficial effect：
1. the present invention obtains yncaria stem with hooks seedling, equipment, medicine and the material that Induction Process uses using the method for multiploid induction
Material is simpler；Because aseptically carrying out, do not influenceed by season and plant growing cycle, it is consistent.Induced efficiency is high, choosing
It is big to select leeway.Because colchicine is induced callus, have while induce tetraploid or the possibility of octoploid, seed selection
Leeway is larger.Shorten the cycle of breeding of new variety.Compared with field routinely using the induction such as infusion process, semar technique, cotton globule leaching method
The time of more than 3 years is saved with the method for seed selection, to meet that the medication demand of people lays the foundation.
2. the yncaria stem with hooks seedling obtained by multiploid induction is moved into seedling shed and cultivated by the present invention, yncaria stem with hooks children is coordinated in seedling shed
The furrow that seedling is formed set up watering system, and the watering tube in watering system can be moved come casting positioning water level by sliding block in slideway
The distance on shower nozzle and ground is adjusted when putting, and watering by electric expansion bar, the water that shower nozzle sprays accurately is injected yncaria stem with hooks
Growing location, and the length at spray end of shower nozzle and the spacing of slideway cooperate, it is ensured that and irrigation amount is uniform, it is ensured that yncaria stem with hooks grows
Water and humidity, root rot illness rate reduces, and the yncaria stem with hooks yield that finally obtains is high, pharmaceutical ingredient content is high.It is in addition, of the invention
The technical scheme of proposition has the characteristics of simple and easy to do, operability and favorable reproducibility.
Further advantage, target and the feature of the present invention embodies part by following explanation, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Brief description of the drawings
Fig. 1 is the structural representation of watering system of the present invention.
Embodiment
With reference to embodiment, the present invention is described in further detail, to make those skilled in the art with reference to specification
Word can be implemented according to this.
It should be appreciated that such as " having ", "comprising" and " comprising " term used herein do not allot one or more
The presence or addition of individual other elements or its combination.It should be noted that experimental method described in following embodiments, such as without spy
Different explanation, is conventional method, the reagent and material, unless otherwise specified, commercially obtains；The present invention's
In description, term " transverse direction ", " longitudinal direction ", " axial direction ", " radial direction ", " on ", " under ", "front", "rear", "left", "right", " vertical ",
The orientation or position relationship of the instruction such as " level ", " top ", " bottom ", " interior ", " outer " are to be closed based on orientation shown in the drawings or position
System, it is for only for ease of and describes of the invention and simplify description, is not that the device of instruction or hint meaning or element must have
Specific orientation, with specific azimuth configuration and operation, therefore be not considered as limiting the invention.
<Embodiment 1>
Step 1: ripe, the full yncaria stem with hooks seed of selection is soaking successively, is transferred in seed culture medium and cultivates after sterilization
20~25d, obtains aseptic seedlings plumular axis, and the seed culture medium is the MS culture mediums without hormone；
Step 2: the plumular axis explant without cotyledonary node that length is 0.4cm is cut from aseptic seedlings plumular axis, by plumular axis
Explant, which is lain in the first callus tissue culture base, carries out light culture, and after 20d, the both ends of plumular axis explant, which are expanded, forms embryo
Axle callus, plumular axis callus is then transferred to progress light culture 20d in the second callus tissue culture base, obtained fresh
Callus, the first callus tissue culture base include MS, 1.0mg/L TDZ, 0.03mg/L NAA, second callus
Tissue culture medium (TCM) includes MS, 1.0mg/L TDZ, 0.03mg/L NAA, 80mg/L colchicine；
Differentiation culture 20d is carried out Step 3: fresh callus is transferred in differential medium, obtains clump bud, it is described
Differential medium includes MS, 0.1% activated carbon；
Strong seedling culture is carried out Step 4: clump bud is transferred in strong seedling culture base, when individual plant seedling length is 3~4cm,
Cut off in basal part of stem, be transferred in root media and carry out culture of rootage, after 20d, grow 1.5~2.0cm root, obtain hook
Rattan seedling, the strong seedling culture base include MS, 1.0mg/L 6-BA, 0.1mg/L NAA, and the root media includes 1/
2MS, 0.5mg/L IBA, 0.5mg/L NAA, 0.2% activated carbon.
The specific method soaked in step 1 is：The yncaria stem with hooks seed chosen is mounted in beanbag, being placed in rotating speed is
4h is soaked on 200rpm shaking table；The specific method of sterilization is：Soaked yncaria stem with hooks seed is placed on superclean bench, used
10% 8~10min of hypochlorite disinfectant, then with sterile water wash 4~6 times.
The cultivation temperature of each step is 20~25 DEG C.
<Embodiment 2>
Step 1: ripe, the full yncaria stem with hooks seed of selection is soaking successively, is transferred in the first culture medium and cultivates after sterilization
20~25d, obtains aseptic seedlings plumular axis, and the seed culture medium is the MS culture mediums without hormone；
Step 2: the plumular axis explant without cotyledonary node that length is 0.6cm is cut from aseptic seedlings plumular axis, by plumular axis
Explant, which is lain in the first callus tissue culture base, carries out light culture, and after 23d, the both ends of plumular axis explant, which are expanded, forms embryo
Axle callus, plumular axis callus is then transferred to progress light culture 15d in the second callus tissue culture base, obtained fresh
Callus, the first callus tissue culture base include MS, 1.2mg/L TDZ, 0.04mg/L NAA, second callus
Tissue culture medium (TCM) includes MS, 1.2mg/L TDZ, 0.04mg/L NAA, 150mg/L colchicine；
Differentiation culture 23d is carried out Step 3: fresh callus is transferred in differential medium, obtains clump bud, it is described
Differential medium include MS, 0.3% activated carbon, 2.0mg/L 6-BA, 0.1mg/L NAA.
Strong seedling culture is carried out Step 4: clump bud is transferred in strong seedling culture base, when individual plant seedling length is 3~4cm,
Cut off in basal part of stem, be transferred in root media and carry out culture of rootage, after 23d, grow 1.5~2.0cm root, obtain hook
Rattan seedling, the strong seedling culture base include MS, 1.3mg/L 6-BA, 0.2mg/L NAA, and the root media includes 1/
2MS, 0.6mg/L IBA, 0.4mg/L NAA, 0.2% activated carbon.
The specific method soaked in step 1 is：The yncaria stem with hooks seed chosen is mounted in beanbag, being placed in rotating speed is
4h is soaked on 200rpm shaking table；The specific method of sterilization is：Soaked yncaria stem with hooks seed is placed on superclean bench, used
10% 8~10min of hypochlorite disinfectant, then with sterile water wash 4~6 times.
The cultivation temperature of each step is 23 DEG C.
<Embodiment 3>
Step 1: ripe, the full yncaria stem with hooks seed of selection is soaking successively, is transferred in the first culture medium and cultivates after sterilization
20~25d, obtains aseptic seedlings plumular axis, and the seed culture medium is the MS culture mediums without hormone；
Step 2: the plumular axis explant without cotyledonary node that length is 0.8cm is cut from aseptic seedlings plumular axis, by plumular axis
Explant, which is lain in the first callus tissue culture base, carries out light culture, and after 25d, the both ends of plumular axis explant, which are expanded, forms embryo
Axle callus, plumular axis callus is then transferred to progress light culture 7d in the second callus tissue culture base, obtained fresh
Callus, the first callus tissue culture base include MS, 1.5mg/L TDZ, 0.05mg/L NAA, second callus
Tissue culture medium (TCM) includes MS, 1.5mg/L TDZ, 0.05mg/L NAA, 200mg/L colchicine；
Differentiation culture 25d is carried out Step 3: fresh callus is transferred in differential medium, obtains clump bud, it is described
Differential medium includes MS, 0.4% activated carbon；
Strong seedling culture is carried out Step 4: clump bud is transferred in strong seedling culture base, when individual plant seedling length is 3~4cm,
Cut off in basal part of stem, be transferred in root media and carry out culture of rootage, after 25d, grow 1.5~2.0cm root, obtain hook
Rattan seedling, the strong seedling culture base include MS, 1.5mg/L 6-BA, 0.2mg/L NAA, and the root media includes 1/
2MS, 0.8mg/L IBA, 0.5mg/L NAA, 0.2% activated carbon.
The specific method soaked in step 1 is：The yncaria stem with hooks seed chosen is mounted in beanbag, being placed in rotating speed is
4h is soaked on 200rpm shaking table；The specific method of sterilization is：Soaked yncaria stem with hooks seed is placed on superclean bench, used
10% 8~10min of hypochlorite disinfectant, then with sterile water wash 4~6 times.
The cultivation temperature of each step is 25 DEG C.
The yncaria stem with hooks seedling that embodiment 1-3 is cultivated is continued in root media to cultivate 7d, then respectively from embodiment 1-3
In randomly select individual plant yncaria stem with hooks seedling, clip tender leaf prepares 10 samples respectively under aseptic conditions, and sets control group, control
The yncaria stem with hooks seedling that group is cultivated for infusion process, infusion process specific method are：First by yncaria stem with hooks seed presoaking and germinating, treat that its radicle shows money or valuables one carries unintentionally
When, yncaria stem with hooks seed is immersed in 24h in 0.5% colchicine solution, elution colchicine is taken out, then sows obtained yncaria stem with hooks
Seedling, clip seedling tender leaf also prepare 10 samples.Then, polyploid detection is carried out using flow cytometer, according to above-mentioned behaviour
Tested three times altogether, three times test result calculations average value, the results are shown in Table 1.
Wherein,
The major pharmaceutical agent and sample treatment of flow cytomery：
(1) medicament
WPB dissociation solutions：0.2mol/L Tris-HC1,4mmol/L MgC12·6H2O, 2mmol/L EDTANa2·
2H2O, 86mmol/L NaCl, 10mmol/L sodium pyrosulfite, 1%PVP-10,1% (7.5,4 DEG C of Triton X-100, pH
Lower preservation.
Fluorescence probe：Propidium iodide (PI), 1mg/ml solution is configured to, is preserved at 4 DEG C.
(2) sample treatment
The fresh leaflet tablet of yncaria stem with hooks seedling is taken respectively, is removed master pulse, is cut 2cm2, it is put into the culture dish of precooling, adds
The WPB dissociation solution 2mL of precooling, disposably quickly shredded with sharp blade, then, WPB dissociation solutions are drawn, with 400 mesh filter membranes
Filtering, collection filtrate, which is placed in 4 DEG C of refrigerators, is incubated 5min, then controls rotating speed to centrifuge 5min for 1000r/min at 4 DEG C.Discard
Supernatant, the WPB dissociation solutions of 200 μ L precoolings are added, while add the PI dyestuffs of 300 μ L precoolings, 4 DEG C of refrigerator is placed in after mixing
In, lucifuge, dyeing 30min is stood, is detected finally to loading pipe.
In addition, by the Duncan Multiple range test methods of SPSS statistics softwares, to embodiment 1-3 inductivity respectively and
The inductivity of control group carries out significant difference inspection, and as a result as shown in table 1, different capitalizations (A, B) represent aobvious 0.01
Work property level error heteropole is notable.
The multiploid induction rate testing result of table 1
| Multiploid induction rate (%) | 0.01 level of signifiance is examined | |
| Embodiment 1 | 48.26 | A |
| Embodiment 2 | 49.96 | A |
| Embodiment 3 | 46.26 | A |
| Control group | 8.23 | B |
By SPSS statistics softwares to 10 samples of every processing, and carry out 3 times and test the multiploid induction respectively obtained
The standard error of rate, the standard error of embodiment 1 is 1.92, and its multiploid induction rate is (48 ± 1.92) %, embodiment 2
Standard error is 1.58, and its multiploid induction rate is (49.96 ± 1.58) %, and the standard error of embodiment 3 is 1.27, its more times
Body inductivity is (46.26 ± 1.27) %, and the standard error of control group is 2.02, its multiploid induction rate for (8.23 ±
2.02) %, the inductivity of the polyploid of embodiment 1 are 5.86 times of control group, and the inductivity of the polyploid of embodiment 2 is control
6.07 times of group, the inductivity of the polyploid of embodiment 3 is 5.62 times of control group.And through significance test, three embodiments
It is extremely notable with the difference of control group.Therefore, the yncaria stem with hooks seedling induced efficiency obtained by the method for the multiploid induction of the present invention
Height, choice are big.
<Embodiment 4>
Step 1: ripe, the full yncaria stem with hooks seed of selection is soaking successively, is transferred in the first culture medium and cultivates after sterilization
20~25d, obtains aseptic seedlings plumular axis, and the seed culture medium is the MS culture mediums without hormone；
Step 2: the plumular axis explant without cotyledonary node that length is 0.6cm is cut from aseptic seedlings plumular axis, by plumular axis
Explant, which is lain in the first callus tissue culture base, carries out light culture, and after 23d, the both ends of plumular axis explant, which are expanded, forms embryo
Axle callus, plumular axis callus is then transferred to progress light culture 15d in the second callus tissue culture base, obtained fresh
Callus, the first callus tissue culture base include MS, 1.2mg/L TDZ, 0.04mg/L NAA, second callus
Tissue culture medium (TCM) includes MS, 1.2mg/L TDZ, 0.04mg/L NAA, 150mg/L colchicine；
Differentiation culture 23d is carried out Step 3: fresh callus is transferred in differential medium, obtains clump bud, it is described
Differential medium include MS, 0.3% activated carbon, 2.0mg/L 6-BA, 0.1mg/L NAA.
Strong seedling culture is carried out Step 4: clump bud is transferred in strong seedling culture base, when individual plant seedling length is 3~4cm,
Cut off in basal part of stem, be transferred in root media and carry out culture of rootage, after 23d, grow 1.5~2.0cm root, obtain hook
Rattan seedling, the strong seedling culture base include MS, 1.3mg/L 6-BA, 0.2mg/L NAA, and the root media includes 1/
2MS, 0.6mg/L IBA, 0.4mg/L NAA, 0.2% activated carbon.
Step 5: taking the blade of the yncaria stem with hooks seedling of step 4 acquisition, pass through flow cytometer and carry out polyploid detection, screening
And reservation is the yncaria stem with hooks seedling of polyploid, i.e. yncaria stem with hooks polyploid seedling, utilizes the strong seedling culture base and root media of step 4
Yncaria stem with hooks polyploid seedling expand numerous.Expand numerous quantity it is as needed depending on.
Step 6: by expand it is numerous after yncaria stem with hooks polyploid seedling replanting to seedling shed in cultivate, be specially：
A, the soil in seedling shed is toasted, then soil deeptillage until soil it is fine and soft, without grume, be sprinkled into animal manure
Just, and humus soil is added, regulation pH is 6.5~8.0, and then laying depth is 5cm sand soil, and is 2m's according to width
Specification sets two row's furrow, and a width of 20cm gutter is provided between two row's furrow；
B, it is 23 DEG C to control seedling temperature of shed, and the land area that 20 × 20cm is taken according to every plant of yncaria stem with hooks polyploid seedling is entered
Row transplanting, after the completion of transplanting, sprinkling root water, the vitamin C of the root water including 7~10 parts by weight, 4~6 parts by weight
Brassinosteroid, the heteroauxin of 3~6 parts by weight, the gibberellin of 2~7 parts by weight, the sodium selenite of 5~9 parts by weight, 4~7
The sodium molybdate of parts by weight, 3~5 parts of compound amino acids, 0.4~0.9 part of garlic selenium polysaccharide, 6~11 parts by weight egg vinegar liquid,
The water of 100 parts by weight mixes；
C, afterwards, every watering in 4 days once, every ventilation in 2 days once, control in seedling shed relative humidity for 80%~
85%.
Watering system is used in step b and step c, when sprinkling root water and watering, as shown in figure 1, described pour water system
System is arranged in the seedling shed, and the watering system includes two water-supply-pipes 11, N number of slideway 12 and 2N and pours water assembly, institute
Two horizontal tops in the seedling shed of the horizontal parallel of water-supply-pipe 11 are stated, and are each perpendicular to the width of the furrow 10, often
N number of apopore 111 is equidistantly provided with along pipe range direction on root water-supply-pipe 11, N number of 12 equidistant horizontal parallel of slideway is set up
Between two water-supply-pipes 11, each slideway 12 is vertical with two water-supply-pipes 11, two ends of a slideway 12
Portion corresponds to described two water-supply-pipes, 11 respective apopores 111, and setting two on a slideway 12 pours water assembly, described
Pour a sliding block 131 that water assembly includes being slidably connected with slideway, be fixedly connected with sliding block 131 electric expansion bar 132 with
And a watering tube 133, the sealing of apopore 111 connection corresponding with one of end of slideway 12 of one end of watering tube 133,
The other end is the shower nozzle 134 that there is strip to spray end being connected with the free end of electric expansion bar 132；
During watering, two sliding blocks 131 in a slideway 12 drive respective watering tube 133 to slide into a wherein row respectively
The both sides of furrow 10, the elongation of electric expansion bar 132 make the shower nozzle 134 be close to yncaria stem with hooks plant, shower nozzle 134 and the distance on ground in seedling shed
15~20cm, the spacing between the adjacent slideway 12 of any two are equal to the length at the strip spray end of the shower nozzle 134.It is actual
In use, the enough sliding block 131 of the length of watering tube 133 drives it to be moved in slideway 12, to prevent watering tube due to gravity
Be hung out in the air, can be arranged multiple loose collars on slideway, watering tube 134 pass through after loose collar respectively with apopore 111 and
Shower nozzle 134 connects.
The specific method soaked in step 1 is：The yncaria stem with hooks seed chosen is mounted in beanbag, being placed in rotating speed is
4h is soaked on 200rpm shaking table；The specific method of sterilization is：Soaked yncaria stem with hooks seed is placed on superclean bench, used
10% 8~10min of hypochlorite disinfectant, then with sterile water wash 4~6 times.
The cultivation temperature of each step is 23 DEG C.
<Comparative example>
The yncaria stem with hooks seedling planted using conventional method, follow-up cultivation is carried out in greenhouse, is directly filled using traditional water pipe
Enter watering, every watering in 4 days once, every ventilation in 2 days once, it is 80%~85% to control relative humidity in greenhouse.
Comparative example is identical with the cultivated area and planting density of the yncaria stem with hooks in embodiment 4.After yncaria stem with hooks harvest, statistics is respective
Hook branch yield, and each ten plants of the yncaria stem with hooks of harvest is randomly choosed from embodiment 4 and comparative example respectively, dries rear grinds, respectively
2.0g is taken, by the content of high effective liquid chromatography for measuring medicinal ingredient (rhynchophyllin and isorhynchophylline), as a result such as table 2 below institute
Show.
The yncaria stem with hooks yield of table 2 and its pharmaceutical ingredient content
| Per mu yield (kg) | Yncaria stem with hooks alkali content (%) | Isorhynchophylline content (%) | The root rot incidence of disease (%) | |
| Embodiment 4 | 484.6 | 0.032 | 0.0065 | 7.3 |
| Comparative example | 317.0 | 0.015 | 0.0037 | 18.6 |
From table 2 it can be seen that forming yncaria stem with hooks seedling by multiploid induction in the embodiment of the present invention 4, the later stage is in seedling shed
By bedding cultivation, and distinctive watering system is used, coordinate rational watering draft type, the yncaria stem with hooks plant yield finally given carries
High 34.7%52.87%, the content of rhynchophyllin and isorhynchophylline is improved in yncaria stem with hooks, under the incidence of disease of root rot is obvious
Drop.
Although embodiment of the present invention is disclosed as above, it is not restricted in specification and embodiment listed
With it can be applied to various suitable the field of the invention completely, can be easily for those skilled in the art
Other modification is realized, therefore under the universal limited without departing substantially from claim and equivalency range, it is of the invention and unlimited
In specific details.
Claims (7)
1. a kind of breeding method of yncaria stem with hooks, it is characterised in that comprise the following steps：
Step 1: 20~25d of culture will be transferred in seed culture medium after yncaria stem with hooks seed successively soaking, sterilization, obtain sterile
Seedling plumular axis；
Step 2: the plumular axis explant without cotyledonary node that length is 0.4~0.8cm is cut from aseptic seedlings plumular axis, by embryo
Axle explant, which is lain in the first callus tissue culture base, carries out light culture, and after 20~25d, the both ends of plumular axis explant are expanded i.e.
Formed plumular axis callus, then by plumular axis callus be transferred in the second callus tissue culture base carry out light culture 7~
20d, obtain fresh callus；
Differentiation 20~25d of culture is carried out Step 3: fresh callus is transferred in differential medium, obtains clump bud；
Strong seedling culture is carried out Step 4: clump bud is transferred in strong seedling culture base, when individual plant seedling length is 3~4cm, in stem
Base portion is cut off, and is transferred in root media and is carried out culture of rootage, after 20~25d, grows 1.5~2.0cm root, obtain hook
Rattan seedling.
2. the breeding method of yncaria stem with hooks as claimed in claim 1, it is characterised in that
The seed culture medium is the MS culture mediums without hormone；
The first callus tissue culture base includes MS, 1.0~1.5mg/L TDZ, 0.03~0.05mg/L NAA；
The second callus tissue culture base include MS, 1.0~1.5mg/L TDZ, 0.03~0.05mg/L NAA, 80~
200mg/L colchicine；
The differential medium includes MS, 0.1~0.4% activated carbon；
The strong seedling culture base includes MS, 1.0~1.5mg/L 6-BA, 0.1~0.2mg/L NAA；
The root media includes 1/2MS, 0.5~0.8mg/L IBA, 0.2~0.5mg/L NAA, 0.2% activity
Charcoal.
3. the breeding method of yncaria stem with hooks as claimed in claim 2, it is characterised in that the specific method soaked in step 1 is：Will
The yncaria stem with hooks seed chosen is mounted in beanbag, is placed on the shaking table that rotating speed is 200rpm and is soaked 4h；The specific method of sterilization is：
Soaked yncaria stem with hooks seed is placed on superclean bench, with 10% 8~10min of hypochlorite disinfectant, then uses sterilized water
Cleaning 4~6 times.
4. the breeding method of yncaria stem with hooks as claimed in claim 3, it is characterised in that differential medium also includes described in step 4
2.0mg/L 6-BA, 0.1mg/L NAA.
5. the breeding method of yncaria stem with hooks as claimed in claim 1, it is characterised in that the cultivation temperature of each step is 20~25
℃。
6. the breeding method of yncaria stem with hooks as claimed in claim 1, it is characterised in that also include：
Step 5: taking the blade for the yncaria stem with hooks seedling that step 4 obtains, polyploid detection is carried out by flow cytometer, screens and protects
Stay be polyploid yncaria stem with hooks seedling, i.e. yncaria stem with hooks polyploid seedling, using the strong seedling culture base and root media of step 4 to hook
Rattan polyploid seedling expand numerous；
Step 6: by expand it is numerous after yncaria stem with hooks polyploid seedling replanting to seedling shed in cultivate, be specially：
A, the soil in seedling shed is toasted, then soil deeptillage until soil it is fine and soft, without grume, be sprinkled into excrement of animals, and
Humus soil is added, regulation pH is 6.5~8.0, and then laying depth is 5cm sand soil, and according to the specification that width is 2m
Two row's furrow are set, a width of 20cm gutter is provided between two row's furrow；
B, it is 23 DEG C to control seedling temperature of shed, and the land area that 20 × 20cm is taken according to every plant of yncaria stem with hooks seedling is transplanted, and is moved
After the completion of cultivation, sprinkling root water, in the vitamin C of the root water including 7~10 parts by weight, the rape element of 4~6 parts by weight
Ester, the heteroauxin of 3~6 parts by weight, the gibberellin of 2~7 parts by weight, the sodium selenite of 5~9 parts by weight, 4~7 parts by weight
Sodium molybdate, 3~5 parts of compound amino acids, 0.4~0.9 part of garlic selenium polysaccharide, the egg vinegar liquid of 6~11 parts by weight, 100 parts by weight
Water mix；
C, afterwards, every watering in 4 days once, every ventilation in 2 days once, it is 80%~85% to control relative humidity in seedling shed.
7. the breeding method of yncaria stem with hooks as claimed in claim 6, it is characterised in that in step b and step c, sprinkling root water and
Watering system is used during watering, the watering system is arranged in the seedling shed, the watering system include two water-supply-pipes,
N number of slideway and 2N pour water assembly, the horizontal top in the seedling shed of two water-supply-pipe horizontal parallels, and vertical
In being equidistantly provided with N number of apopore, N number of equidistant water of slideway along pipe range direction on the width of furrow, every water-supply-pipe
Average row is erected between two water-supply-pipes, and each slideway is vertical with two water-supply-pipes, two of a slideway
End corresponds to the respective apopore of two water-supply-pipes, and setting two on a slideway pours water assembly, the watering
Component include be slidably connected with slideway a sliding block, be fixedly connected with a slide block an electric expansion bar and a watering tube,
Corresponding with one of end of the slideway apopore sealing connection of one end of watering tube, the other end be with electric expansion bar from
By the shower nozzle with strip spray end of end connection；
During watering, two sliding blocks in a slideway drive respective watering tube to slide into the both sides of wherein row's furrow respectively, electricity
Dynamic expansion link elongation makes shower nozzle, and close to yncaria stem with hooks plant, shower nozzle and the distance on ground in seedling shed are 15~20cm, and any two is adjacent
Spacing between slideway is equal to the length at the strip spray end of the shower nozzle.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711122593.9A CN107873516A (en) | 2017-11-14 | 2017-11-14 | The breeding method of yncaria stem with hooks |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711122593.9A CN107873516A (en) | 2017-11-14 | 2017-11-14 | The breeding method of yncaria stem with hooks |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107873516A true CN107873516A (en) | 2018-04-06 |
Family
ID=61776637
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711122593.9A Pending CN107873516A (en) | 2017-11-14 | 2017-11-14 | The breeding method of yncaria stem with hooks |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107873516A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109588246A (en) * | 2018-12-29 | 2019-04-09 | 丁年生 | A kind of method for culturing seedlings of horseshoe |
| CN110100582A (en) * | 2018-11-13 | 2019-08-09 | 广西壮族自治区林业科学研究院 | A kind of breeding method shining rattan container seedling |
| CN111165358A (en) * | 2020-03-13 | 2020-05-19 | 临沧惠康中药材资源开发有限公司 | Method for inducing tetraploid plant by using adventitious bud of uncaria |
| CN112005884A (en) * | 2020-09-14 | 2020-12-01 | 广西壮族自治区药用植物园 | Method for improving alkaloid content of subprostrate sophora tissue culture seedling by utilizing selenium |
| CN112154914A (en) * | 2020-10-09 | 2021-01-01 | 广西壮族自治区药用植物园 | Method for improving alkaloid content of subprostrate sophora tissue culture seedling by using calcium ions |
| CN116941528A (en) * | 2023-07-12 | 2023-10-27 | 广西壮族自治区药用植物园 | Method for in-vitro preservation of uncaria |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2011083235A (en) * | 2009-10-16 | 2011-04-28 | Kinjirushi Kk | Method for forming multiple shoot from culture seedling and method for producing virus-free seed yam |
| CN102301952A (en) * | 2011-07-19 | 2012-01-04 | 北京林业大学 | Method for breeding chamomile |
| CN104957043A (en) * | 2015-07-30 | 2015-10-07 | 广西壮族自治区药用植物园 | Rapid propagation method of Uncaria sessilifructus Roxb. |
| CN105993951A (en) * | 2016-05-27 | 2016-10-12 | 陈思 | Tissue-culture rapid-breeding method for stewartia sinensis |
| CN106258976A (en) * | 2016-08-24 | 2017-01-04 | 华中农业大学 | A kind of tissue culturing fast seedling-cultivating method of mustard type rape |
| CN106900492A (en) * | 2017-04-24 | 2017-06-30 | 成都飞航沛腾科技有限公司 | A kind of agricultural irrigation device |
-
2017
- 2017-11-14 CN CN201711122593.9A patent/CN107873516A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2011083235A (en) * | 2009-10-16 | 2011-04-28 | Kinjirushi Kk | Method for forming multiple shoot from culture seedling and method for producing virus-free seed yam |
| CN102301952A (en) * | 2011-07-19 | 2012-01-04 | 北京林业大学 | Method for breeding chamomile |
| CN104957043A (en) * | 2015-07-30 | 2015-10-07 | 广西壮族自治区药用植物园 | Rapid propagation method of Uncaria sessilifructus Roxb. |
| CN105993951A (en) * | 2016-05-27 | 2016-10-12 | 陈思 | Tissue-culture rapid-breeding method for stewartia sinensis |
| CN106258976A (en) * | 2016-08-24 | 2017-01-04 | 华中农业大学 | A kind of tissue culturing fast seedling-cultivating method of mustard type rape |
| CN106900492A (en) * | 2017-04-24 | 2017-06-30 | 成都飞航沛腾科技有限公司 | A kind of agricultural irrigation device |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110100582A (en) * | 2018-11-13 | 2019-08-09 | 广西壮族自治区林业科学研究院 | A kind of breeding method shining rattan container seedling |
| CN110100582B (en) * | 2018-11-13 | 2021-12-14 | 广西壮族自治区林业科学研究院 | Cultivation method of gloryvine container seedlings |
| CN109588246A (en) * | 2018-12-29 | 2019-04-09 | 丁年生 | A kind of method for culturing seedlings of horseshoe |
| CN111165358A (en) * | 2020-03-13 | 2020-05-19 | 临沧惠康中药材资源开发有限公司 | Method for inducing tetraploid plant by using adventitious bud of uncaria |
| CN112005884A (en) * | 2020-09-14 | 2020-12-01 | 广西壮族自治区药用植物园 | Method for improving alkaloid content of subprostrate sophora tissue culture seedling by utilizing selenium |
| CN112005884B (en) * | 2020-09-14 | 2022-02-22 | 广西壮族自治区药用植物园 | Method for improving alkaloid content of subprostrate sophora tissue culture seedling by utilizing selenium |
| CN112154914A (en) * | 2020-10-09 | 2021-01-01 | 广西壮族自治区药用植物园 | Method for improving alkaloid content of subprostrate sophora tissue culture seedling by using calcium ions |
| CN116941528A (en) * | 2023-07-12 | 2023-10-27 | 广西壮族自治区药用植物园 | Method for in-vitro preservation of uncaria |
| CN116941528B (en) * | 2023-07-12 | 2024-04-09 | 广西壮族自治区药用植物园 | Method for in-vitro preservation of uncaria |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107873516A (en) | The breeding method of yncaria stem with hooks | |
| CN105340747B (en) | A kind of Radix Glycyrrhizae asexual rapid propagation method | |
| CN105613199B (en) | A kind of method for culturing seedlings of tea tree | |
| CN103190347B (en) | Teapot dates tissue culturing method | |
| CN107173222A (en) | A kind of wild bletilla striata quick breeding method for tissue culture | |
| CN105309317B (en) | The tissue culture propagation of Flemingia macrophylla | |
| CN105103715A (en) | Bud forcing treatment method for watermelon seeds with high germination rate | |
| CN104542307B (en) | Culturing method of momordica cochinchinensis | |
| CN105918121A (en) | Method for rapidly breeding seedlings by using Giantleaf Ardisia leaves | |
| CN105918133A (en) | Tissue culture and rapid propagation method of euryodendron excelsum | |
| CN103229642A (en) | Breeding method for improved variety of Lonicera japonica Thunb No.1 | |
| CN105993952A (en) | Rapid breeding method of Euryodendron excelsum cultivation seedlings | |
| CN106342614A (en) | Production method of selenium-enriched Mayang longevity fragrant rice | |
| CN106305427A (en) | Tissue culture and rapid propagation method for Acer ginnala | |
| CN103688856A (en) | Rapid propagation method of Blumea riparia (Bl.) DC medicinal material | |
| CN108901858A (en) | A kind of phoenix head ginger selenium-rich original silkworm egg quick-breeding method | |
| CN106165648B (en) | A kind of cercis tissue culture culture medium and cultural method | |
| CN105432466B (en) | Method with plant regeneration occurs for a kind of pittosporum tobira somatic embryo | |
| KR100476422B1 (en) | Seedling production of Polygonatum odoratum and its rhizome processing development for large-scale cultivation | |
| CN111602486A (en) | Seedling transplanting method for canna edulis ker seedling seeds | |
| CN104585040A (en) | Method for quickly propagating germchit of giantleaf ardisia rhizome through bud plumular axis | |
| CN108739403A (en) | A kind of tissue culture and rapid propagation method of rose wood | |
| CN107743868A (en) | A kind of method for efficiently breeding roxburgh anoectochilus terminal bud using nature optical culture forming seedling through one step culture | |
| CN104054578B (en) | The tissue culture and rapid propagation method of Dian clove | |
| CN106688556A (en) | High-yield cultivation method of aralia elata seem in barren land |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |