CN107182780A - The method for culturing seedlings of P. kingianum - Google Patents
The method for culturing seedlings of P. kingianum Download PDFInfo
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- CN107182780A CN107182780A CN201710290369.4A CN201710290369A CN107182780A CN 107182780 A CN107182780 A CN 107182780A CN 201710290369 A CN201710290369 A CN 201710290369A CN 107182780 A CN107182780 A CN 107182780A
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- 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
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- 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
Abstract
The method for culturing seedlings of P. kingianum, step includes：P. kingianum explant prepares, and eliminates browning culture, and callus induction, adventitious bud occur and Multiplying culture, promote the light culture of root growth, test tube seedling rejuvenation culture, culture of rootage, hardening culture；Beneficial effect is：Yunnan gold explant carries out the switching culture every 7 days in culture medium A, removes the browning effect occurred during tissue culture notable；Callus induction, clump bud are carried out simultaneously to occur and Multiplying culture, improve reproductive efficiency, breeding coefficient can bring up to more than 8.67, and the tissue culture time can foreshorten to 45 d；Light culture stage, the development degree of P. kingianum rhizome is greatly improved, and survival rate is up to more than 95%；The present invention is stage by stage using four kinds of culture medium switching cultures, overcome the problem of easily growing fungi and accumulation virus when P. kingianum carries out tissue culture, turn out the seedling adaptability come and resistance is extremely strong, reduce tissue culture seedling pollution rate, improve the stability of Different Varieties.
Description
Technical field
The invention belongs to Chinese medicine tissue culture technology field, it is related to a kind of method for culturing seedlings of P. kingianum.
Background technology
P. kingianum（Polygonatum kingianum Coll.er Hemsl.）For Liliaceae（Liliaceae）Polygonatum
（Polygonatum）Herbaceos perennial, mainly originates in the ground such as the river ﹑ Guizhou of Yun Nan ﹑ tetra- and Vietnam, the rare distribution of Burma；It is raw
In on underbrush or dark and damp careless slope, rock.P. kingianum is traditional Chinese medicine, and its medicinal part is root-like stock, its mild-natured, taste
Sweet, for nourishing top grade, primary efficacy is nourishing Yin and moistening dryness, tonifying kidney and benefiting sperm, for weakness of the spleen and the stomach, fatigue and asthenia, dry deficiency of food, the deficiency syndrome of the lung
Cough caused by dryness, asthenia of essence and blood, Heat Diabetes.Modern pharmacology research shows that also there is P. kingianum immune stimulating, enhancing to be immunized, delay to decline
Always, antiviral the effects such as, there are bright prospects in terms of research new drug and exploitation health products.Due to P. kingianum have it is important medicinal
And edibility, the huge market demand, wild P. kingianum often excavate by being ransacked of property, and resource is on the verge of exhaustion, the exploitation to P. kingianum
Extremely disadvantageous influence is brought using with sustainable development.
The breeding of P. kingianum has two kinds of generative propagation and vegetative propagation.In generative propagation, P. kingianum seed has variation
Greatly, the features such as diversity height, strong adaptability, but its development is slower, and aerial part 1 year of needs, growth are grown from being seeded into
And harvest cycle is long.The vegetative propagation of P. kingianum is bred by rhizome, and this mode is more universal in various regions use, but
Fungi and accumulation virus are easily grown in the rhizome for being long-term vegetative propagation P. kingianum, when carrying out tissue-cultured seedling, outside P. kingianum
The easy browning of implant, tissue-cultured seedling is easily contaminated, and this can cause P. kingianum reproduction speed slow, and planting percent is low, what Different Varieties were degenerated
Problem, it is impossible to provide a large amount of high quality seedlings, constrains the artificial large area plantation of P. kingianum.
At present, it is domestic to have many reports on some kind Study on tissue culture of Polygonatum, but do not refer to that explant is easy
The problem of browning and pollution, and its breeding coefficient is relatively low, per a generation, breeding is needed 60-90 days, growing-seedling period length, test tube seedling transition are tamed and dociled
Survival rate is low during change so that the industrialization production of P. kingianum seedling is difficult to realize with current method for tissue culture.
The content of the invention
There is the breeding such as slow reproduction speed, easy browning, cultivation cycle length, rhizome for the breeding of P. kingianum in the prior art
Fungi and accumulation virus are easily grown in body, the problem of Different Varieties are degenerated is caused, the present invention provides a kind of nursery of P. kingianum
Method, concrete scheme comprises the following steps：
（1）P. kingianum explant prepares：In annual spring, eugonic P. kingianum rhizome is dug out from soil, washed away with hairbrush
Soil, then cuts band young shoot rhizome with scalpel, soaks 10-15 min for 20% detergent solution with mass ratio, puts stream
30-60 min are rinsed under water；It is put into after flushing in the beaker on superclean bench, it is 20% vitamin C that mass ratio is added in beaker
Soak 5-10 min；Then soaked with volume ratio for 75% ethanol after 30 s, be transferred in the mercury chloride that mass ratio is 0.1% and sterilize
5-8 min, aseptic water washing 8-10 times is not less than 3 min every time, obtains P. kingianum explant, standby；
（2）Eliminate browning culture：Ready Yunnan gold explant is inoculated in culture medium A, is 1500-2000 in illuminance
Lx, light application time is 8 h/d, and temperature is is cultivated under conditions of 23 ± 1 DEG C, every culture medium A more renewed for seven days continues
Culture, the replacing number of times of culture medium A is no less than twice；
Culture medium A is that 5000-10000 mg polyethylene is added in every liter of distilled water than lattice alkanoic acid ketone, 1.0-3.0 g work
Property charcoal, 1.0-10 mg sodium chloride, 15000 mg sucrose and 5300-5500 mg agar powder are formulated, culture medium A
PH value is between 5.4-5.8；
（3）Callus induction, adventitious bud occur and Multiplying culture：The P. kingianum explant after browning will be eliminated and access culture medium
In B, illuminance be 1500-2000 lx, light application time be 12 h/d, temperature be 23 ± 1 DEG C under conditions of synchronously carry out
Callus induction, adventitious bud occur and Multiplying culture；The bud for cultivating P. kingianum explant after 15 d starts sprouting and grows blade,
And there is callus in base portion, the more rate that obtains is 91.23%；Callus surface starts indefinite clump bud occur after 30 d, indefinite clump
The differentiation rate that adventitious bud clump is differentiated in bud can reach 100%；45 d metaplexus inductivities are up to 5.37；Culture medium B is by every liter
1.5-2.0 mg zeatin, 0.05-0.1 mg methyl α-naphthyl acetate, 30000 mg sucrose, 5300-5500 are added in distilled water
Mg agar is formulated, and culture medium B pH value is between 5.4-5.8；
（4）Promote the light culture of root growth：Adventitious bud clump induce, healthy and strong is divided into 2-3 strains one clump, is forwarded to new
In fresh culture medium B, light culture, callus proliferation, bud are carried out under conditions of 18 ± 1 DEG C in no light, temperature control
While a length of clump bud of growing thickly, base portion rhizome increases, and the flourishing shape albefaction test tube seedling of growing thickly of rhizome can be obtained after 30 d；
（5）Test tube seedling rejuvenation culture：By the flourishing albefaction test tube seedling band rhizome of rhizome in culture medium B be cut into 0.5 cm ×
0.5 cm sizes are transferred in culture medium C；It is 1500-2000 lx in illuminance, light application time is 12 h/d, and temperature is 23 ± 1
Cultivated under conditions of DEG C, small rhizome is mushroomed out after 30 d of culture, while clump bud amount reproduction；It can be grown after 45 d
Healthy and strong test tube seedling, its breeding coefficient is up to 8.67；
Culture medium C is the zeatin that 1.5-2.0 mg are added in every liter of distilled water, 0.05-0.1 mg methyl α-naphthyl acetate, 5-10 mg
Paclobutrazol, 30000 mg sucrose, 5300-5500 mg agar is formulated, the pH value of culture medium C 5.4-5.8 it
Between；
（6）Culture of rootage：The test tube seedling of a height of 3-4 cm length is inoculated in culture medium D, is 1500-2000 lx in illuminance,
Light application time is 12 h/d, and temperature obtains taking root for growing way stalwartness to be cultivated under conditions of 23 ± 1 DEG C after culture 45d
Seedling；
Culture medium D is the methyl α-naphthyl acetate that 0.5-1.5 mg are added in every liter of distilled water, 15000 mg sucrose, 5300-5500 mg
Agar be formulated, culture medium D pH value is between 5.4-5.8；
（7）The high rooted seedlings of 6-8 cm are placed in after the d of room temperature lower refining seedling 3, rooted seedling is taken out from culture medium, are cleaned up residual
The culture medium stayed in rooted seedling, is put into mass concentration 0.1-0.2% carbendazim solution（Mass ratio）Middle sterilization 2-3 min, it is false
Put in sand bed, keep air humidity more than 90%, sheltered from heat or light with 70% sunshade net, survival rate more than 95% after 30 d；Later every
10 d can transplant crop field after spraying the MS nutrient solutions of 1 20 times of dilution, 30 d.
It is using the beneficial effect of the technical program：
The 1st, Yunnan gold explant is inoculated in the continuous switching culture carried out in culture medium A every 7 days, Yunnan can be preferably removed yellow
The browning occurred during smart Explants In Tissue Culture.
2nd, in culture medium B, occur and Multiplying culture while carrying out callus induction, clump bud, simplify culture program,
Reproductive efficiency is improved, breeding coefficient can bring up to more than 8.67, and the tissue culture time can foreshorten to 45 d.Cultivated in culture medium B
There is callus in the base portion of P. kingianum explant after 15 d, and the more rate that obtains can reach 91.23%；Callus surface is opened after 30 d
The differentiation rate for beginning to occur differentiating in indefinite clump bud, indefinite clump bud adventitious bud clump can reach 100%；45 d metaplexus inductivities can
Up to 5.37.
Adventitious bud clump refer to callus surface it is differentiated go out bud point；Indefinite clump bud then refer to clump bud grow to naked eyes can
See the young shape of young plant；There is the callus of adventitious bud clump in clump bud differentiation rate=the callus number of the indefinite clump bud of appearance/
Number.
3rd, the light culture stage in seedling raising process, the development degree of P. kingianum rhizome is greatly improved, it is ensured that transplanting into
Motility rate, survival rate is up to more than 95%.
4th, culture processes of the present invention are relatively simple, stage by stage using four kinds of culture medium switching cultures, overcome P. kingianum and enter
The problem of fungi and accumulation virus are easily grown during row tissue culture, turns out the seedling adaptability come and resistance is extremely strong, drop
Low tissue culture seedling pollution rate, improves the stability of Different Varieties.
5th, whole year production can be achieved well in culturing room with tissue culture technique, has both saved land resource, improved again
Economic benefit, overcomes the difficult point that traditional modes of reproduction can not be produced in the anniversary.
Embodiment
Embodiment 1：A kind of method for culturing seedlings of P. kingianum, comprises the following steps：
（1）P. kingianum explant prepares：In annual spring, eugonic P. kingianum rhizome is dug out from soil, washed away with hairbrush
Soil, then cuts band young shoot rhizome with scalpel, soaks 10 min for 20% detergent solution with mass ratio, puts flowing water
30 min of lower flushing；It is put into after flushing in the beaker on superclean bench, it is 20% vitamin C immersion 5 that mass ratio is added in beaker
min；Then soaked with volume ratio for 75% ethanol after 30 s, be transferred in the mercury chloride that mass ratio is 0.1% and sterilize 5 min, nothing
Bacterium water is rinsed 8 times, is every time 4 min, is obtained P. kingianum explant, standby；
（2）Eliminate browning culture：Ready Yunnan gold explant is inoculated in culture medium A, is 1500 lx in illuminance,
Light application time is 8 h/d, and temperature is is cultivated under conditions of 22 DEG C, every culture medium A more renewed for seven days continues to cultivate, training
Base A replacing number of times is supported no less than twice；
Culture medium A is the polyethylene that 5000 mg are added in every liter of distilled water than lattice alkanoic acid ketone, 1.0 g activated carbon, 1.0 mg
Sodium chloride, 15000 mg sucrose and 5300 mg agar powder is formulated, and the pH value of culture medium A is 5.4；
（3）Callus induction, adventitious bud occur and Multiplying culture：The P. kingianum explant after browning will be eliminated and access culture medium
It is 1500 lx in illuminance, light application time is 12 h/d, and temperature lures synchronously to carry out callus under conditions of 22 DEG C in B
Lead, adventitious bud occurs and Multiplying culture；The bud for cultivating P. kingianum explant after 15 d starts sprouting and grows blade, and goes out in base portion
Existing callus, the more rate that obtains is 91.23%；Callus surface starts indefinite clump bud occur after 30 d, is differentiated in indefinite clump bud
The differentiation rate of adventitious bud clump can reach 100%；45 d metaplexus inductivities are up to 5.37；Culture medium B is added in every liter of distilled water
Enter 1.5 mg zeatin, 0.05 mg methyl α-naphthyl acetate, 30000 mg sucrose, 5300 mg agar is formulated, culture medium
B pH value is 5.4；
（4）Promote the light culture of root growth：Adventitious bud clump induce, healthy and strong is divided into 2 plants one clump, is forwarded to fresh
Culture medium B in, carry out light culture under conditions of 17 DEG C in no light, temperature control, callus proliferation, bud are grown thickly length
While for clump bud, base portion rhizome increases, and the flourishing shape albefaction test tube seedling of growing thickly of rhizome can be obtained after 30 d；
（5）Test tube seedling rejuvenation culture：By the flourishing albefaction test tube seedling band rhizome of rhizome in culture medium B be cut into 0.5 cm ×
0.5 cm sizes are transferred in culture medium C；It is 1500 lx in illuminance, light application time is 12 h/d, and temperature is 22 DEG C of condition
Lower to be cultivated, small rhizome is mushroomed out after 30 d of culture, while clump bud amount reproduction；The examination of robust growth can be obtained after 45 d
Guan Miao, its breeding coefficient is up to 8.67；
Culture medium C is the zeatin that 1.5 mg are added in every liter of distilled water, 0.05 mg methyl α-naphthyl acetate, 5 mg paclobutrazol,
30000 mg sucrose, 5300 mg agar is formulated, and the pH value of culture medium C is 5.4；
（6）Culture of rootage：The test tube seedling of a height of 3 cm length is inoculated in culture medium D, is 1500 lx in illuminance, during illumination
Between be 12 h/d, temperature be 22 DEG C under conditions of cultivated, culture 45 d after obtain growing way stalwartness rooted seedling；
Culture medium D is the methyl α-naphthyl acetate that 0.5 mg is added in every liter of distilled water, and 15000 mg sucrose, 5300 mg agar is matched somebody with somebody
System is formed, and culture medium D pH value is 5.4；
（7）The high rooted seedlings of 6 cm are placed in after the d of room temperature lower refining seedling 3, rooted seedling is taken out from culture medium, residual is cleaned up
Culture medium in rooted seedling, is put into the carbendazim solution of mass concentration 0.1%（Mass ratio）2 min of middle sterilization, vacation is put in sand bed
On, air humidity more than 90% is kept, is sheltered from heat or light with 70% sunshade net, survival rate more than 95% after 30 d；Sprayed later every 10 d
Crop field can be transplanted after the MS nutrient solutions of 1 20 times of dilution, 30 d.
Embodiment 2：A kind of method for culturing seedlings of P. kingianum, comprises the following steps：
（1）P. kingianum explant prepares：In annual spring, eugonic P. kingianum rhizome is dug out from soil, washed away with hairbrush
Soil, then cuts band young shoot rhizome with scalpel, soaks 15 min for 20% detergent solution with mass ratio, puts flowing water
60 min of lower flushing；It is put into after flushing in the beaker on superclean bench, mass ratio is added in beaker and is soaked for 20% vitamin C
10 min；Then soaked with volume ratio for 75% ethanol after 30 s, be transferred in the mercury chloride that mass ratio is 0.1% and sterilize 8 min,
Aseptic water washing 10 times, is every time 5 min, obtains P. kingianum explant, standby；
（2）Eliminate browning culture：Ready Yunnan gold explant is inoculated in culture medium A, is 2000 lx in illuminance,
Light application time is 8 h/d, and temperature is is cultivated under conditions of 24 DEG C, every culture medium A more renewed for seven days continues to cultivate, training
Base A replacing number of times is supported no less than twice；
Culture medium A is the polyethylene that 10000 mg are added in every liter of distilled water than lattice alkanoic acid ketone, 3.0 g activated carbon, 10 mg
Sodium chloride, 15000 mg sucrose and 5500 mg agar powder is formulated, and the pH value of culture medium A is 5.8；
（3）Callus induction, adventitious bud occur and Multiplying culture：The P. kingianum explant after browning will be eliminated and access culture medium
It is 2000 lx in illuminance, light application time is 12 h/d, and temperature lures synchronously to carry out callus under conditions of 24 DEG C in B
Lead, adventitious bud occurs and Multiplying culture；The bud for cultivating P. kingianum explant after 15 d starts sprouting and grows blade, and goes out in base portion
Existing callus, the more rate that obtains is 91.23%；Callus surface starts indefinite clump bud occur after 30 d, is differentiated in indefinite clump bud
The differentiation rate of adventitious bud clump can reach 100%；45 d metaplexus inductivities are up to 5.37；Culture medium B is added in every liter of distilled water
Enter 2.0 mg zeatin, 0.1 mg methyl α-naphthyl acetate, 30000 mg sucrose, 5500 mg agar is formulated, culture medium B
PH value 5.8；
（4）Promote the light culture of root growth：Adventitious bud clump induce, healthy and strong is divided into 3 plants one clump, is forwarded to fresh
Culture medium B in, carry out light culture under conditions of 19 DEG C in no light, temperature control, callus proliferation, bud are grown thickly length
While for clump bud, base portion rhizome increases, and the flourishing shape albefaction test tube seedling of growing thickly of rhizome can be obtained after 30 d；
（5）Test tube seedling rejuvenation culture：By the flourishing albefaction test tube seedling band rhizome of rhizome in culture medium B be cut into 0.5 cm ×
0.5 cm sizes are transferred in culture medium C；It is 2000 lx in illuminance, light application time is 12 h/d, and temperature is 24 DEG C of condition
Lower to be cultivated, small rhizome is mushroomed out after 30 d of culture, while clump bud amount reproduction；The examination of robust growth can be obtained after 45 d
Guan Miao, its breeding coefficient is up to 8.67；
Culture medium C is the zeatin that 2.0 mg are added in every liter of distilled water, 0.1 mg methyl α-naphthyl acetate, 10 mg paclobutrazol,
30000 mg sucrose, 5500 mg agar is formulated, and the pH value of culture medium C is 5.8；
（6）Culture of rootage：The test tube seedling of a height of 4 cm length is inoculated in culture medium D, is 2000 lx in illuminance, during illumination
Between be 12 h/d, temperature be 24 DEG C under conditions of cultivated, culture 45 d after obtain growing way stalwartness rooted seedling；
Culture medium D is the methyl α-naphthyl acetate that 1.5 mg are added in every liter of distilled water, and 15000 mg sucrose, 5500 mg agar is matched somebody with somebody
System is formed, and culture medium D pH value is 5.8；
（7）The high rooted seedlings of 8 cm are placed in after the d of room temperature lower refining seedling 3, rooted seedling is taken out from culture medium, residual is cleaned up
Culture medium in rooted seedling, is put into the carbendazim solution of mass concentration 0.2%（Mass ratio）3 min of middle sterilization, vacation is put in sand bed
On, air humidity more than 90% is kept, is sheltered from heat or light with 70% sunshade net, survival rate more than 95% after 30 d；Sprayed later every 10 d
Crop field can be transplanted after the MS nutrient solutions of 1 20 times of dilution, 30 d.
Claims (1)
1. the method for culturing seedlings of P. kingianum, it is characterised in that concrete scheme comprises the following steps：
（1）P. kingianum explant prepares：In annual spring, eugonic P. kingianum rhizome is dug out from soil, washed away with hairbrush
Soil, then cuts band young shoot rhizome with scalpel, soaks 10-15 min for 20% detergent solution with mass ratio, puts stream
30-60 min are rinsed under water；It is put into after flushing in the beaker on superclean bench, it is 20% vitamin C that mass ratio is added in beaker
Soak 5-10 min；Then soaked with volume ratio for 75% ethanol after 30 s, be transferred in the mercury chloride that mass ratio is 0.1% and sterilize
5-8 min, aseptic water washing 8-10 times is not less than 3 min every time, obtains P. kingianum explant, standby；
（2）Eliminate browning culture：Ready Yunnan gold explant is inoculated in culture medium A, is 1500-2000 in illuminance
Lx, light application time is 8 h/d, and temperature is is cultivated under conditions of 23 ± 1 DEG C, every culture medium A more renewed for seven days continues
Culture, the replacing number of times of culture medium A is no less than twice；
Culture medium A is that 5000-10000 mg polyethylene is added in every liter of distilled water than lattice alkanoic acid ketone, 1.0-3.0 g work
Property charcoal, 1.0-10 mg sodium chloride, 15000 mg sucrose and 5300-5500 mg agar powder are formulated, culture medium A
PH value is between 5.4-5.8；
（3）Callus induction, adventitious bud occur and Multiplying culture：The P. kingianum explant after browning will be eliminated and access culture medium
In B, illuminance be 1500-2000 lx, light application time be 12 h/d, temperature be 23 ± 1 DEG C under conditions of synchronously carry out
Callus induction, adventitious bud occur and Multiplying culture；The bud for cultivating P. kingianum explant after 15 d starts sprouting and grows blade,
And there is callus in base portion；Callus surface starts indefinite clump bud occur after 30 d；45 d metaplexus inductivities are reachable
5.37；
Culture medium B is the zeatin that 1.5-2.0 mg are added in every liter of distilled water, 0.05-0.1 mg methyl α-naphthyl acetate, 30000
Mg sucrose, 5300-5500 mg agar is formulated, and culture medium B pH value is between 5.4-5.8；
（4）Promote the light culture of root growth：Adventitious bud clump induce, healthy and strong is divided into 2-3 strains one clump, is forwarded to new
In fresh culture medium B, light culture, callus proliferation, bud are carried out under conditions of 18 ± 1 DEG C in no light, temperature control
While a length of clump bud of growing thickly, base portion rhizome increases, and the flourishing shape albefaction test tube seedling of growing thickly of rhizome can be obtained after 30 d；
（5）Test tube seedling rejuvenation culture：By the flourishing albefaction test tube seedling band rhizome of rhizome in culture medium B be cut into 0.5 cm ×
0.5 cm sizes are transferred in culture medium C；It is 1500-2000 lx in illuminance, light application time is 12 h/d, and temperature is 23 ± 1
Cultivated under conditions of DEG C, small rhizome is mushroomed out after 30 d of culture, while clump bud amount reproduction；It can be grown after 45 d
Healthy and strong test tube seedling；
Culture medium C is the zeatin that 1.5-2.0 mg are added in every liter of distilled water, 0.05-0.1 mg methyl α-naphthyl acetate, 5-10 mg
Paclobutrazol, 30000 mg sucrose, 5300-5500 mg agar is formulated, the pH value of culture medium C 5.4-5.8 it
Between；
（6）Culture of rootage：The test tube seedling of a height of 3-4 cm length is inoculated in culture medium D, is 1500-2000 lx in illuminance,
Light application time is 12 h/d, and temperature obtains taking root for growing way stalwartness to be cultivated under conditions of 23 ± 1 DEG C after culture 45d
Seedling；
Culture medium D is the methyl α-naphthyl acetate that 0.5-1.5 mg are added in every liter of distilled water, 15000 mg sucrose, 5300-5500 mg
Agar be formulated, culture medium D pH value is between 5.4-5.8；
（7）The high rooted seedlings of 6-8 cm are placed in after the d of room temperature lower refining seedling 3, rooted seedling is taken out from culture medium, are cleaned up residual
The culture medium stayed in rooted seedling, is put into mass concentration 0.1-0.2% carbendazim solution（Mass ratio）Middle sterilization 2-3 min, it is false
Put in sand bed, keep air humidity more than 90%, sheltered from heat or light with 70% sunshade net, survival rate more than 95% after 30 d；Later every
10 d can transplant crop field after spraying the MS nutrient solutions of 1 20 times of dilution, 30 d.
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| CN114041410A (en) * | 2021-11-10 | 2022-02-15 | 山东省农业科学院 | Rapid seedling raising method for polygonatum kingianum seeds |
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| CN110463609B (en) * | 2019-09-11 | 2021-05-07 | 云南中医药大学 | Method for improving artificial rapid propagation efficiency by overcoming tip necrosis of Sesamum indicum |
| CN110463609A (en) * | 2019-09-11 | 2019-11-19 | 云南中医药大学 | The method that bigflower centranthera root overcomes burned tip necrosis to improve artificial micropropagation efficiency |
| CN112042521A (en) * | 2020-08-10 | 2020-12-08 | 阳山县三连阳生态农林开发有限公司 | Transplanting method for test-tube plantlet of rhizome plant |
| CN111955346A (en) * | 2020-09-03 | 2020-11-20 | 云南华农农业有限公司 | Novel method for inhibiting browning and improving artificial rapid propagation efficiency of Monte raspberries |
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| CN114041410A (en) * | 2021-11-10 | 2022-02-15 | 山东省农业科学院 | Rapid seedling raising method for polygonatum kingianum seeds |
| CN114041410B (en) * | 2021-11-10 | 2022-06-24 | 山东省农业科学院 | Rapid seedling raising method for polygonatum kingianum seeds |
| CN113994862A (en) * | 2021-11-19 | 2022-02-01 | 吉首大学 | Flowering phase regulation and control method for dendrobium moniliforme |
| CN116508652A (en) * | 2023-04-26 | 2023-08-01 | 陇东学院 | Clone establishment and micro-rhizome rapid propagation method for polygonatum sibiricum |
| CN116508652B (en) * | 2023-04-26 | 2024-03-08 | 陇东学院 | Clone establishment and micro-rhizome rapid propagation method for polygonatum sibiricum |
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