CN106577301A - Method of industrially cultivating protocorm of dendrobium officinale without hormones - Google Patents
Method of industrially cultivating protocorm of dendrobium officinale without hormones Download PDFInfo
- Publication number
- CN106577301A CN106577301A CN201710056436.6A CN201710056436A CN106577301A CN 106577301 A CN106577301 A CN 106577301A CN 201710056436 A CN201710056436 A CN 201710056436A CN 106577301 A CN106577301 A CN 106577301A
- Authority
- CN
- China
- Prior art keywords
- protocorm
- culture
- medium
- hormone
- herba dendrobii
- 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.)
- Granted
Links
- 229940088597 hormone Drugs 0.000 title claims abstract description 41
- 239000005556 hormone Substances 0.000 title claims abstract description 41
- 238000000034 method Methods 0.000 title claims abstract description 16
- 241001076416 Dendrobium tosaense Species 0.000 title abstract description 6
- 230000006698 induction Effects 0.000 claims abstract description 6
- 239000001963 growth medium Substances 0.000 claims description 52
- 239000002609 medium Substances 0.000 claims description 52
- 239000012530 fluid Substances 0.000 claims description 37
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 26
- 229930006000 Sucrose Natural products 0.000 claims description 26
- 239000005720 sucrose Substances 0.000 claims description 26
- 239000001888 Peptone Substances 0.000 claims description 20
- 108010080698 Peptones Proteins 0.000 claims description 20
- 238000005286 illumination Methods 0.000 claims description 20
- 239000002054 inoculum Substances 0.000 claims description 20
- 235000019319 peptone Nutrition 0.000 claims description 20
- 239000007787 solid Substances 0.000 claims description 17
- 241000026010 Dendrobium candidum Species 0.000 claims description 15
- 229920001525 carrageenan Polymers 0.000 claims description 15
- 235000010418 carrageenan Nutrition 0.000 claims description 15
- 229920001817 Agar Polymers 0.000 claims description 14
- 239000008272 agar Substances 0.000 claims description 14
- 241000196324 Embryophyta Species 0.000 claims description 6
- 239000002994 raw material Substances 0.000 claims description 6
- 238000005273 aeration Methods 0.000 claims description 5
- 235000013399 edible fruits Nutrition 0.000 claims description 5
- 230000035800 maturation Effects 0.000 claims description 5
- 230000010355 oscillation Effects 0.000 claims description 5
- 230000008117 seed development Effects 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 2
- 239000010977 jade Substances 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 abstract description 6
- 230000035755 proliferation Effects 0.000 abstract 2
- 238000009776 industrial production Methods 0.000 abstract 1
- 238000007689 inspection Methods 0.000 abstract 1
- 238000009331 sowing Methods 0.000 abstract 1
- 240000000111 Saccharum officinarum Species 0.000 description 4
- 235000007201 Saccharum officinarum Nutrition 0.000 description 4
- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical class CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 description 4
- 229940066779 peptones Drugs 0.000 description 4
- AIMMVWOEOZMVMS-UHFFFAOYSA-N cyclopropanecarboxamide Chemical compound NC(=O)C1CC1 AIMMVWOEOZMVMS-UHFFFAOYSA-N 0.000 description 3
- 244000046052 Phaseolus vulgaris Species 0.000 description 2
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 230000035784 germination Effects 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 241001523681 Dendrobium Species 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 206010020649 Hyperkeratosis Diseases 0.000 description 1
- 241000234295 Musa Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 206010030899 opisthotonus Diseases 0.000 description 1
- 210000004681 ovum Anatomy 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
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
- A01H4/008—Methods for regeneration to complete plants
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Mushroom Cultivation (AREA)
Abstract
The invention relates to a method of industrially cultivating protocorm of dendrobium officinale without hormones and belongs to the technical field of hormone-free cultivation. The hormone-free industrially cultivated dendrobium officinale is obtained by steps of sowing of dendrobium officinale, induction of protocorm, proliferation of protocorm and output of protocorm. The proliferation multiplying power of the protocorm is greatly improved and the stem yield thereof is improved, so that industrial production of the protocorm of dendrobium officinale has actual application value. As a hormone-free production mode is adopted, the obtained product meets related inspection requirements and can satisfy the demand in the market.
Description
Technical field
The present invention relates to a kind of method for cultivating Herba Dendrobii protocorm without hormone industrialization, belongs to without hormone culture technique
Field.
Background technology
Herba Dendrobii（Scientific name：Dendrobium officinaleKimura et Migo）:Stem is upright, cylinder, long
9-35 centimetre, thick 2-4 millimeters, branch, does not have more piece；Ye Erlie, papery, long round shape lanceolar, edge and middle rib are often with pale purple
Color.Raceme often sends from the old stem top of the leaf that fallen, and has 2-3 flower；Petal piece dry film matter, shallow white is avette, long 5-7
Millimeter, sepal and petal yellow green, nearly similar, long round shape lanceolar, lip white, base portion has the calluss of 1 green or yellow
Body, ovum shape lanceolar is more slightly shorter than sepal, middle part opisthotonos.Stamen post yellow green, is about 3 millimeters, and tip both sides respectively have 1 purple point；Medicine
Cap white, long ovate triangle is about 2.3 millimeters, and the nearly sharp point in top and 2 are split.The month at florescence 3-6.
Herba Dendrobii protocorm has been proved to containing the effective nutritional labeling similar with the fresh stem of Herba Dendrobii, Ke Yizuo
Raw material is extracted in enrichment for vegetable polysaccharidess.But for a long time Herba Dendrobii protocorm all simply does in research institutions' laboratory
Research is used, or plant factor as seedlings of Dendrobium officinale breeding plumule, no matter all remote from production scale or yield rate
It is far from reaching the requirement of industrialized production.
During protocorm dry product is made in fresh protocorm drying, there is one " rate must be done " can be used as protocorm stem
The reference of quality.Must do that rate is higher, yield rate is higher, and relative quality is also better.Common laboratory uses pure solid
Culture medium prepares protocorm, its must do rate only have in 3 ~ 5%, and preparation process be easy to differentiation germination, affect again subculture increase
Grow.
If preparing protocorm using neat liquid culture medium, its must do rate can than slightly higher the 2 ~ 3% of the preparation of solid culture,
But in subculture for several times, the growth rate of protocorm can be reduced by generation, protocorm granule it is bigger than normal, middle easy in inactivation is obtained
Dry rate also progressively declines.
The content of the invention
It is an object of the invention to overcome above-mentioned weak point, there is provided a kind of to cultivate Herba Dendrobii protocorm without hormone industrialization
The method of stem, what the technique substantially increased the propagation multiplying power of protocorm and improve it must do rate, make Herba Dendrobii protocorm
The industrialized production of stem has the value of practical application.
According to the technical scheme that the present invention is provided, a kind of method for cultivating Herba Dendrobii protocorm without hormone industrialization, step
It is rapid as follows：
（1）Herba Dendrobii is sowed：Using Herba Dendrobii maturation Fruit pod as the induction raw material of protocorm, Seeds of Dendrobium Candidum is broadcast
Plant on without hormone culture-medium G0, seed tiling culture base plane；Condition of culture be 20 ~ 28 DEG C of temperature, illumination 2000 ~
3000Lx, cultivates 12 ~ 15 days, the Seeds of Dendrobium Candidum after being expanded；
（2）Protocorm is induced：By step（1）Seeds of Dendrobium Candidum after gained expands is inoculated in fluid medium G1, is inoculated with
Amount is 8 ~ 10 grams of every 200mL；It is placed on shaking table and uniformly vibrates, frequency of oscillation is 80 ~ 110 revs/min, keeping temperature 20 ~ 28
DEG C, 2000 ~ 3000Lx of illumination, once, after subculture 2 ~ 3 times, seed development is protocorm to 15 days subcultures of culture；
（3）Protocorm Multiplication：
A, by step（2）Protocorm in gained fluid medium G1 is transferred in solid medium G2 to be cultivated, and inoculum concentration is every
8 ~ 10 grams of 200mL；Condition is 20 ~ 28 DEG C of temperature, and 2000 ~ 3000Lx of illumination is cultivated 30 ~ 45 days；
B, by step a gained solid medium G2 in protocorm be transferred in fluid medium G3 cultivate, inoculum concentration be 30 ~
50 grams/600mL bottles；Condition is that temperature is kept for 20 ~ 28 DEG C, and 2000 ~ 3000Lx of illumination, shaking speed is 80 ~ 100 revs/min；30
In ~ 45 days follow-up generations, are once；
（4）Protocorm is produced：By step（3）Gained fluid medium G3 in protocorm be seeded to produce before without hormone culture
In base G4, inoculum concentration is 8 ~ 10 grams of every 200mL；During the air chamber bucket that volume is 18L is forwarded to after 30 days, bucket is trained built with liquid
Foster base G5,14L, inoculum concentration is 1200 ~ 1500 grams/barrel, continues to be filled with pure air into bucket during culture, aeration quantity is 0.2 ~
0.25 cube m/h；To full bottle after cultivating 30 days, can directly obtain without hormone Herba Dendrobii protocorm product.
Step（1）Described is 1/2MS culture medium+90 ~ 110g/L Rhizoma Solani tuber osi+20g/L sugarcanes without hormone culture-medium G0 concrete components
Sugar+5 ~ 6g/L carrageenans/agar, pH6.0.
Step（2）The fluid medium G1 be 1/2MS culture medium+0.2 ~ 0.5mg/L 6-BA+10g/L Fructus Musaes+20 ~
25g/L sucrose, pH6.2；
Step（3）The solid medium G2 is：1/2MS culture medium+0.2 ~ 0.5mg/L 6-BA+0.20 ~ 0.3g/L peptones+
100g/L Rhizoma Solani tuber osi+20 ~ 25g/L sucrose+5 ~ 6g/L carrageenans/agar, pH6.2；
Fluid medium G3 formula is 1/2MS culture medium+0.20 ~ 0.3g/L peptone+100g/L Rhizoma Solani tuber osi+20 ~ 25g/L sucrose,
pH6.2。
Step（4）It is described without hormone culture-medium G4 be 1/2MS culture medium+0.20 ~ 0.3g/L peptone+100g/L Rhizoma Solani tuber osis+
20 ~ 25g/L sucrose+5 ~ 6g/L carrageenans/agar, pH6.2；
The fluid medium G5 formula is 1/2MS culture medium+0.25 ~ 0.3g/L peptone+100g/L Rhizoma Solani tuber osi+25g/L sucrose,
pH6.2。
Beneficial effects of the present invention：What the present invention substantially increased the propagation multiplying power of protocorm and improve it must do rate,
Making the industrialized production of Herba Dendrobii protocorm has the value of practical application.Because the present invention adopts the producer without hormone
Formula, therefore the product for obtaining meets related check requirement, can reach listing requirement.
Specific embodiment
Embodiment 1
A kind of method for cultivating Herba Dendrobii protocorm without hormone industrialization, step is as follows：
（1）Herba Dendrobii is sowed：Using Herba Dendrobii maturation Fruit pod as the induction raw material of protocorm, Seeds of Dendrobium Candidum is broadcast
Plant on without hormone culture-medium G0, seed tiling culture base plane, culture medium is about 200mL, height 2cm, the very thin paving of seed
One layer；Condition of culture is 20 DEG C of temperature, and illumination 3000Lx is cultivated 12 days, the Seeds of Dendrobium Candidum after being expanded；
（2）Protocorm is induced：By step（1）Seeds of Dendrobium Candidum after gained expands is inoculated in fluid medium G1, is inoculated with
Amount is 8 grams of every 200mL；It is placed on shaking table and uniformly vibrates, frequency of oscillation is 80 revs/min, 20 DEG C of keeping temperature, illumination
2000Lx, once, after subculture 2 times, seed development is protocorm to 15 days subcultures of culture；
（3）Protocorm Multiplication：
A, by step（2）Protocorm in gained fluid medium G1 is transferred in solid medium G2 to be cultivated, and inoculum concentration is every
8 grams of 200mL；Condition is 20 DEG C of temperature, and illumination 2000Lx is cultivated 30 days；
B, by step a gained solid medium G2 in protocorm be transferred in fluid medium G3 cultivate, inoculum concentration is 30
Gram/600mL bottles；Condition is that temperature is kept for 20 DEG C, and illumination 2000Lx, shaking speed is 80 revs/min；In 30 days follow-up generations, are once；
（4）Protocorm is produced：By step（3）Gained fluid medium G3 in protocorm be seeded to produce before without hormone culture
In base G4, inoculum concentration is 8 grams of every 200mL；During the air chamber bucket that volume is 18L is forwarded to after 30 days, bucket is built with fluid medium
G5,14L, inoculum concentration is 1200 grams/barrel, continues to be filled with pure air into bucket during culture, aeration quantity is 0.2 cubic metre/it is little
When；To full bottle after cultivating 30 days, can directly obtain without hormone Herba Dendrobii protocorm product.
Step（1）It is described without hormone culture-medium G0 concrete components be 1/2MS culture medium+90g/L Rhizoma Solani tuber osi+20g/L sucrose+
5g/L carrageenans/agar, pH6.0.
Step（2）The fluid medium G1 is 1/2MS culture medium+0.2mg/L 6-BA+10g/L Fructus Musae+20g/L sugarcanes
Sugar, pH6.2；
Step（3）The solid medium G2 is：1/2MS culture medium+0.2mg/L 6-BA+0.20 ~ 0.3g/L peptones+
100g/L Rhizoma Solani tuber osi+20g/L sucrose+5g/L carrageenans/agar, pH6.2；
Fluid medium G3 formula be 1/2MS culture medium+0.20g/L peptone+100g/L Rhizoma Solani tuber osi+20g/L sucrose, pH6.2.
Step（4）Described is 1/2MS culture medium+0.20g/L peptone+100g/L Rhizoma Solani tuber osi+20g/L without hormone culture-medium G4
Sucrose+5g/L carrageenans/agar, pH6.2；
The fluid medium G5 formula is 1/2MS culture medium+0.25g/L peptone+100g/L Rhizoma Solani tuber osi+25g/L sucrose,
pH6.2。
Embodiment 2
A kind of method for cultivating Herba Dendrobii protocorm without hormone industrialization, step is as follows：
（1）Herba Dendrobii is sowed：Using Herba Dendrobii maturation Fruit pod as the induction raw material of protocorm, Seeds of Dendrobium Candidum is broadcast
Plant on without hormone culture-medium G0, seed tiling culture base plane, culture medium is about 200mL, height 2cm, the very thin paving of seed
One layer；Condition of culture is 28 DEG C of temperature, and illumination 3000Lx is cultivated 15 days, the Seeds of Dendrobium Candidum after being expanded；
（2）Protocorm is induced：By step（1）Seeds of Dendrobium Candidum after gained expands is inoculated in fluid medium G1, is inoculated with
Amount is 10 grams of every 200mL；It is placed on shaking table and uniformly vibrates, frequency of oscillation is 110 revs/min, 28 DEG C of keeping temperature, illumination
3000Lx, once, after subculture 3 times, seed development is protocorm to 15 days subcultures of culture；
（3）Protocorm Multiplication：
A, by step（2）Protocorm in gained fluid medium G1 is transferred in solid medium G2 to be cultivated, and inoculum concentration is every
10 grams of 200mL；Condition is 28 DEG C of temperature, and illumination 3000Lx is cultivated 45 days；
B, by step a gained solid medium G2 in protocorm be transferred in fluid medium G3 cultivate, inoculum concentration is 50
Gram/600mL bottles；Condition is that temperature is kept for 28 DEG C, and illumination 3000Lx, shaking speed is 100 revs/min；45 days follow-up generations one
It is secondary；
（4）Protocorm is produced：By step（3）Gained fluid medium G3 in protocorm be seeded to produce before without hormone culture
In base G4, inoculum concentration is 10 grams of every 200mL；During the air chamber bucket that volume is 18L is forwarded to after 30 days, bucket is built with liquid culture
Base G5,14L, inoculum concentration is 1500 grams/barrel, continues to be filled with pure air into bucket during culture, aeration quantity is 0.25 cubic metre/
Hour；To full bottle after cultivating 30 days, can directly obtain without hormone Herba Dendrobii protocorm product.
Step（1）It is described without hormone culture-medium G0 concrete components be 1/2MS culture medium+110g/L Rhizoma Solani tuber osi+20g/L sucrose+
6g/L carrageenans/agar, pH6.0.
Step（2）The fluid medium G1 is 1/2MS culture medium+0.5mg/L 6-BA+10g/L Fructus Musae+25g/L sugarcanes
Sugar, pH6.2；
Step（3）The solid medium G2 is：1/2MS culture medium+0.5mg/L 6-BA+0.3g/L peptones+100g/L are native
Bean+25g/L sucrose+6g/L carrageenans/agar, pH6.2；
Fluid medium G3 formula be 1/2MS culture medium+0.3g/L peptone+100g/L Rhizoma Solani tuber osi+25g/L sucrose, pH6.2.
Step（4）Described is 1/2MS culture medium+0.3g/L peptone+100g/L Rhizoma Solani tuber osi+25g/L without hormone culture-medium G4
Sucrose+6g/L carrageenans/agar, pH6.2；
The fluid medium G5 formula is 1/2MS culture medium+0.3g/L peptone+100g/L Rhizoma Solani tuber osi+25g/L sucrose,
pH6.2。
Embodiment 3
A kind of method for cultivating Herba Dendrobii protocorm without hormone industrialization, step is as follows：
（1）Herba Dendrobii is sowed：Using Herba Dendrobii maturation Fruit pod as the induction raw material of protocorm, Seeds of Dendrobium Candidum is broadcast
Plant on without hormone culture-medium G0, seed tiling culture base plane, culture medium is about 200mL, height 2cm, the very thin paving of seed
One layer；Condition of culture is 24 DEG C of temperature, and illumination 2500Lx is cultivated 14 days, the Seeds of Dendrobium Candidum after being expanded；
（2）Protocorm is induced：By step（1）Seeds of Dendrobium Candidum after gained expands is inoculated in fluid medium G1, is inoculated with
Amount is 9 grams of every 200mL；It is placed on shaking table and uniformly vibrates, frequency of oscillation is 100 revs/min, 25 DEG C of keeping temperature, illumination
2500Lx, once, after subculture 2 times, seed development is protocorm to 15 days subcultures of culture；
（3）Protocorm Multiplication：
A, by step（2）Protocorm in gained fluid medium G1 is transferred in solid medium G2 to be cultivated, and inoculum concentration is every
9 grams of 200mL；Condition is 24 DEG C of temperature, and illumination 2500Lx is cultivated 40 days；
B, by step a gained solid medium G2 in protocorm be transferred in fluid medium G3 cultivate, inoculum concentration is 40
Gram/600mL bottles；Condition is that temperature is kept for 25 DEG C, and illumination 2500Lx, shaking speed is 90 revs/min；In 35 days follow-up generations, are once；
（4）Protocorm is produced：By step（3）Gained fluid medium G3 in protocorm be seeded to produce before without hormone culture
In base G4, inoculum concentration is every 200mL9 gram；During the air chamber bucket that volume is 18L is forwarded to after 30 days, bucket is built with fluid medium
G5,14L, inoculum concentration is 1350 grams/barrel, continues to be filled with pure air into bucket during culture, aeration quantity is 0.22 cubic metre/it is little
When；To full bottle after cultivating 30 days, can directly obtain without hormone Herba Dendrobii protocorm product.
Step（1）It is described without hormone culture-medium G0 concrete components be 1/2MS culture medium+100g/L Rhizoma Solani tuber osi+20g/L sucrose+
6g/L carrageenans/agar, pH6.0.
Step（2）The fluid medium G1 is 1/2MS culture medium+0.4mg/L 6-BA+10g/L Fructus Musae+24g/L sugarcanes
Sugar, pH6.2；
Step（3）The solid medium G2 is：1/2MS culture medium+0.4mg/L 6-BA+0.25g/L peptones+100g/L are native
Bean+23g/L sucrose+5 ~ 6g/L carrageenans/agar, pH6.2；
Fluid medium G3 formula be 1/2MS culture medium+0.25g/L peptone+100g/L Rhizoma Solani tuber osi+23g/L sucrose, pH6.2.
Step（4）It is described without hormone culture-medium G4 be 1/2MS culture medium+0.20 ~ 0.3g/L peptone+100g/L Rhizoma Solani tuber osis+
23g/L sucrose+6g/L carrageenans/agar, pH6.2；
The fluid medium G5 formula is 1/2MS culture medium+0.28g/L peptone+100g/L Rhizoma Solani tuber osi+25g/L sucrose,
pH6.2。
Four steps by more than, Herba Dendrobii protocorm gross weight can reach 2560 times of initial inducing amount, and obtaining dry rate can be with
Reach 10% or so.Its polyoses content and non-oxidizability can reach the level of the fresh stem of high-quality Herba Dendrobii, even better than fresh stem.
The alternate mode of mushy stage culture medium that the present invention is used, can simultaneously solve asking in the above-mentioned two mode of production
Topic, can keep the vigor of protocorm so as to be truly realized hundred generation subcultures and keep stable biological nature, will not occur in advance
Differentiation germination phenomenon.Simultaneously as the nutrient substance of abundance is with the addition of in culture medium of the present invention, so when 30 ~ 36 days,
Protocorm can reach the summit that must do rate, and close 13%.
Claims (5)
1. a kind of method for cultivating Herba Dendrobii protocorm without hormone industrialization, is characterized in that step is as follows：
（1）Herba Dendrobii is sowed：Using Herba Dendrobii maturation Fruit pod as the induction raw material of protocorm, Seeds of Dendrobium Candidum is broadcast
Plant on without hormone culture-medium G0, seed tiling culture base plane；Condition of culture be 20 ~ 28 DEG C of temperature, illumination 2000 ~
3000Lx, cultivates 12 ~ 15 days, the Seeds of Dendrobium Candidum after being expanded；
（2）Protocorm is induced：By step（1）Seeds of Dendrobium Candidum after gained expands is inoculated in fluid medium G1, is inoculated with
Amount is 8 ~ 10 grams of every 200mL；It is placed on shaking table and uniformly vibrates, frequency of oscillation is 80 ~ 110 revs/min, keeping temperature 20 ~ 28
DEG C, 2000 ~ 3000Lx of illumination, once, after subculture 2 ~ 3 times, seed development is protocorm to 15 days subcultures of culture；
（3）Protocorm Multiplication：
A, by step（2）Protocorm in gained fluid medium G1 is transferred in solid medium G2 to be cultivated, and inoculum concentration is every
8 ~ 10 grams of 200mL；Condition is 20 ~ 28 DEG C of temperature, and 2000 ~ 3000Lx of illumination is cultivated 30 ~ 45 days；
B, by step a gained solid medium G2 in protocorm be transferred in fluid medium G3 cultivate, inoculum concentration be 30 ~
50 grams/600mL bottles；Condition is that temperature is kept for 20 ~ 28 DEG C, and 2000 ~ 3000Lx of illumination, shaking speed is 80 ~ 100 revs/min；30
In ~ 45 days follow-up generations, are once；
（4）Protocorm is produced：By step（3）Gained fluid medium G3 in protocorm be seeded to produce before without hormone culture
In base G4, inoculum concentration is 8 ~ 10 grams of every 200mL；During the air chamber bucket that volume is 18L is forwarded to after 30 days, bucket is trained built with liquid
Foster base G5,14L, inoculum concentration is 1200 ~ 1500 grams/barrel, continues to be filled with pure air into bucket during culture, aeration quantity is 0.2 ~
0.25 cube m/h；To full bottle after cultivating 30 days, can directly obtain without hormone Herba Dendrobii protocorm product.
2. the method for cultivating Herba Dendrobii protocorm without hormone industrialization as claimed in claim 1, is characterized in that：Step（1）Institute
It is 1/2MS culture medium+90 ~ 110g/L Rhizoma Solani tuber osi+20g/L sucrose+5 ~ 6g/L carrageenans/fine jade to state without hormone culture-medium G0 concrete components
Fat, pH6.0.
3. the method for cultivating Herba Dendrobii protocorm without hormone industrialization as claimed in claim 1, is characterized in that：Step（2）Institute
Fluid medium G1 is stated for 1/2MS culture medium+0.2 ~ 0.5mg/L 6-BA+10g/L Fructus Musae+20 ~ 25g/L sucrose, pH6.2.
4. the method for cultivating Herba Dendrobii protocorm without hormone industrialization as claimed in claim 1, is characterized in that：Step（3）Institute
Stating solid medium G2 is：1/2MS culture medium+0.2 ~ 0.5mg/L 6-BA+0.20 ~ 0.3g/L peptone+100g/L Rhizoma Solani tuber osis+20
~ 25g/L sucrose+5 ~ 6g/L carrageenans/agar, pH6.2；
Fluid medium G3 formula is 1/2MS culture medium+0.20 ~ 0.3g/L peptone+100g/L Rhizoma Solani tuber osi+20 ~ 25g/L sucrose,
pH6.2。
5. the method for cultivating Herba Dendrobii protocorm without hormone industrialization as claimed in claim 1, is characterized in that：Step（4）Institute
It is 1/2MS culture medium+0.20 ~ 0.3g/L peptone+100g/L Rhizoma Solani tuber osi+20 ~ 25g/L+5 ~ 6g/ of sucrose to state without hormone culture-medium G4
L carrageenans/agar, pH6.2；
The fluid medium G5 formula is 1/2MS culture medium+0.25 ~ 0.3g/L peptone+100g/L Rhizoma Solani tuber osi+25g/L sucrose,
pH6.2。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710056436.6A CN106577301B (en) | 2017-01-25 | 2017-01-25 | A kind of method that dendrobium candidum protocorm is cultivated in no hormone industrialization |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710056436.6A CN106577301B (en) | 2017-01-25 | 2017-01-25 | A kind of method that dendrobium candidum protocorm is cultivated in no hormone industrialization |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106577301A true CN106577301A (en) | 2017-04-26 |
| CN106577301B CN106577301B (en) | 2018-06-01 |
Family
ID=58585238
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710056436.6A Expired - Fee Related CN106577301B (en) | 2017-01-25 | 2017-01-25 | A kind of method that dendrobium candidum protocorm is cultivated in no hormone industrialization |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106577301B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114376133A (en) * | 2022-01-18 | 2022-04-22 | 常州九葆生物科技有限公司 | Dendrobium officinale primary pulp beverage prepared based on protocorm and preparation method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101768566A (en) * | 2010-01-25 | 2010-07-07 | 大连普瑞康生物技术有限公司 | Method for culturing cendrobium officinale protocorm by intermittent fermentation |
| CN104067940A (en) * | 2014-06-30 | 2014-10-01 | 广州花都先锋园艺有限公司 | Hormone-free tissue culture method for dendrobium officinale |
| CN104705188A (en) * | 2015-04-03 | 2015-06-17 | 南京工业大学 | method for culturing dendrobium officinale protocorm |
| KR20150103879A (en) * | 2014-03-04 | 2015-09-14 | 고려대학교 산학협력단 | Producing method of orchid seedlings |
-
2017
- 2017-01-25 CN CN201710056436.6A patent/CN106577301B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101768566A (en) * | 2010-01-25 | 2010-07-07 | 大连普瑞康生物技术有限公司 | Method for culturing cendrobium officinale protocorm by intermittent fermentation |
| KR20150103879A (en) * | 2014-03-04 | 2015-09-14 | 고려대학교 산학협력단 | Producing method of orchid seedlings |
| CN104067940A (en) * | 2014-06-30 | 2014-10-01 | 广州花都先锋园艺有限公司 | Hormone-free tissue culture method for dendrobium officinale |
| CN104705188A (en) * | 2015-04-03 | 2015-06-17 | 南京工业大学 | method for culturing dendrobium officinale protocorm |
Non-Patent Citations (1)
| Title |
|---|
| 余乐等: "铁皮石斛离体快繁技术", 《江苏农业科学》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114376133A (en) * | 2022-01-18 | 2022-04-22 | 常州九葆生物科技有限公司 | Dendrobium officinale primary pulp beverage prepared based on protocorm and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106577301B (en) | 2018-06-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1954668B (en) | Scale tissue culture quickly reproducing method of saffron crocus seedball (sprout) | |
| CN103210842B (en) | Dendrobium candidum test-tube flowering and seed setting method | |
| CN108401794A (en) | A kind of armillaria mellea accreting with Rhizoma Gastrodiae liquid spawn production method and cultigen special culture media | |
| CN109337895A (en) | A kind of production method of good quality and high output selenium-enriched hericium erinaceus strain | |
| CN105441340A (en) | Gibberellin GA4+7High-yield strain and application thereof | |
| CN103650912B (en) | A kind of multi-spore inbred breeding method for Flammulina velutipes | |
| CN105567572A (en) | Preparation technology of agaricus bisporus strain | |
| CN107950288B (en) | Straw mushroom cultivation process | |
| CN105519345B (en) | The gastrodia elata sexual propagation method that cotton seed hulls is that major ingredient makes Germination Strain production kind | |
| CN107333651A (en) | A kind of method for improving Dendrobidium huoshanness protocorm alkaloid | |
| CN108541513A (en) | A kind of winter sweet-smelling grass liquid spawn quick-breeding method | |
| CN106577301B (en) | A kind of method that dendrobium candidum protocorm is cultivated in no hormone industrialization | |
| CN105838621B (en) | A kind of culture solution and breeding method of grifola frondosus liquid spawn | |
| CN107873392A (en) | A kind of potted plant three-dimensional Industry Cultivation integration system of elegant precious mushroom and application | |
| CN101984787B (en) | Method for strengthening tissue culture seedlings of rhododendron lapponicum | |
| CN107018905A (en) | A kind of blue test tube seedling preserving seed method of bottle | |
| CN116472916A (en) | Method for obtaining golden fungus fruiting body by mixed strain | |
| KR20130098946A (en) | A method for cultivating gastrodia elata seeds | |
| CN103503771A (en) | Tissue culture and rapid propagation method for Australian hardenbergia violacea seedlings | |
| CN104145714A (en) | Effective breeding method for king trumpet mushroom production | |
| CN108029553A (en) | Biological engineering's method of rice dry measure used in former times | |
| CN111264303B (en) | Lyophyllum fumosoroseum cultivar and preparation method thereof | |
| CN101401515B (en) | Cymbidium hybridum tissue culture seedling contamination cultivation method | |
| CN107494269A (en) | A kind of elimination little Hua all ages in blue tissue culture procedures endophyte method | |
| CN104137776B (en) | Blueberry tissue cultured seedling liquid rooting method and special culturing container |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180601 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |