CN106538384B - A kind of seedling method for test tube propagation of width leaf thickness lip orchid - Google Patents
A kind of seedling method for test tube propagation of width leaf thickness lip orchid Download PDFInfo
- Publication number
- CN106538384B CN106538384B CN201610935238.2A CN201610935238A CN106538384B CN 106538384 B CN106538384 B CN 106538384B CN 201610935238 A CN201610935238 A CN 201610935238A CN 106538384 B CN106538384 B CN 106538384B
- Authority
- CN
- China
- Prior art keywords
- seedling
- test tube
- leaf thickness
- thickness lip
- culture
- 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.)
- Expired - Fee Related
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 seedling method for test tube propagation of wide leaf thickness lip orchid, include the following steps：Step 1：The wide leaf thickness lip orchid of acquisition is developed to ripe and uncracked fruit as explant；Step 2：Powdered embryo in explant is subjected to axenic germination and obtains protocorm, the constituent content of culture medium used by the axenic germination is：It is 5.4~5.8 that 0.5~2.0g/L, which spends No. 1 precious, 0.2~1.0mg/L NAA, 15~30g/L sucrose, 3.5~6.0g/L agar, 50~100ml/L coconut milk, pH,；Step 3：Protocorm Multiplication is obtained into protocorms；Step 4：Protocorms differentiation culture is obtained into seedling；Step 5：Seedling culture is obtained into test tube seedling；Step 6：By test tube transplantation of seedlings up to seedling.The object of the present invention is to provide a kind of seedling method for test tube propagation of wide leaf thickness lip orchid, the advantage of this method is that, very high germination rate can be kept high.
Description
Technical field
The present invention relates to the method for Plant Tissue Breeding in agricultural biotechnologies more particularly to a kind of kinds of wide leaf thickness lip orchid
Seedling method for test tube propagation.
Background technology
Wide leaf thickness lip blue (Epigeneium amplum (Lindl.) Summerh.) belongs to orchid family thickness lip epidendrum, is
The maximum kind of individual plants in Chinese producing region in belonging to, sepal and petal apex are suddenly tapering, the nearly diamond shape of sliver in lip, thereon
Face is thick lip orchid species more easy to identify without gusset.Wide leaf thickness lip orchid is distributed mainly on the ground such as Yunnan Province of China, Tibet and Guangxi,
Be born in the hayashishita or small stream side rock of 1000~1900 meters of height above sea level and mountainous region woods on trunk.In ethnic groups such as Yunnan, Tibet
Area, wide leaf thickness lip orchid are commonly used for treating the diseases such as bronchitis, encephalitis, pulmonary tuberculosis, hepatitis, dysentery, chronic osteomyelitis.By
In the extensive pharmacological action of wide leaf thickness lip orchid, people excessively excavate wide leaf thickness lip orchid, in addition its wild environment is by tight
It destroys again, its wild resource is caused to fall sharply, distribution is drastically reduced, therefore there is an urgent need to carry out artificial breeding to wide leaf thickness lip orchid
It educates.
Currently, wide leaf thickness lip orchid species seedling is mainly bred by plant division mode, but there are reproductive efficiencies low, of high cost, period
Long problem cannot be satisfied the demand of wide leaf thickness lip orchid scale Commercial Growers seedling.In addition, wide leaf thickness lip orchid species is due to embryo
Ateliosis, under natural conditions extremely difficult sprouting.
Invention content
The object of the present invention is to provide a kind of seedling method for test tube propagation of wide leaf thickness lip orchid, the advantage of this method is that,
It can keep very high germination rate high.
The technical scheme is that：A kind of seedling method for test tube propagation of width leaf thickness lip orchid, includes the following steps：
Step 1：The wide leaf thickness lip orchid of acquisition is developed to ripe and uncracked fruit as explant；
Step 2：Powdered embryo in explant is subjected to axenic germination and obtains protocorm, the axenic germination is used
The constituent content of culture medium be：0.5~2.0g/L spend precious No. 1,0.2~1.0mg/L NAA, 15~30g/L sucrose, 3.5~
6.0g/L agar, 50~100ml/L coconut milk, pH are 5.4~5.8；
Step 3：Protocorm Multiplication is obtained into protocorms；
Step 4：Protocorms differentiation culture is obtained into seedling；
Step 5：Seedling culture is obtained into test tube seedling；
Step 6：By test tube transplantation of seedlings up to seedling.
In the seedling method for test tube propagation of above-mentioned wide leaf thickness lip orchid, in the step 2, the powder in explant
The acquisition methods of shape embryo are：Explant is impregnated 1~5 minute in ethyl alcohol, in disinfection 10~30 after rinsed with sterile water 3~5 times
Minute, 5~7 incision fruits of rinsed with sterile water obtain powdered embryo；
For specifically, explant is impregnated 1~5 minute in the ethyl alcohol of 75vol%, after rinsed with sterile water 3~5 times
It is sterilized 10~30 minutes in 0.1% mercuric chloride solution, 5~7 incision fruits of rinsed with sterile water obtain powdered embryo.
In the seedling method for test tube propagation of above-mentioned wide leaf thickness lip orchid, in the step 2, powdered embryo is in culture medium
Middle incubation time is 90~120 days.
In the seedling method for test tube propagation of above-mentioned wide leaf thickness lip orchid, the group of used culture medium is proliferated in step 3
It is divided into：Improve Knudson C, 0.1~1.0mg/L 6-BA, 0.01~0.3mg/L TDZ, 15~30g/L sucrose, 3.5~
6.0g/L agar, 50~100ml/L coconut milk, 0.2~0.5g/L activated carbons, pH are 5.4~5.8.
It should be noted that improvement Knudson C are culture mediums commonly used in the art, can specifically refer to《Yunnan plant is ground
Study carefully》6 (2), 224,1984, article name is《The influence of plant hormone and culture medium to gastrodia seed germination》, author's section gold and jade.
In the seedling method for test tube propagation of above-mentioned wide leaf thickness lip orchid, incubation time in the step 3 is 30~
45 days, growth coefficient was 3~5 times.
In the seedling method for test tube propagation of above-mentioned wide leaf thickness lip orchid, in the step 4, break up culture medium used
Group be divided into：Improve Knudson C, 0.1~1.0mg/L NAA, 15~30g/L sucrose, 3.5~6.0g/L agar, 50~
100ml/L coconut milk, 0.2~0.5g/L activated carbons, pH are 5.4~5.8.
In the seedling method for test tube propagation of above-mentioned wide leaf thickness lip orchid, in the step 4, differentiation culture it is used when
Between be 40~60 days.
In the seedling method for test tube propagation of above-mentioned wide leaf thickness lip orchid, the component of the culture medium used in the step 5
For：0.1~1.0g/L spend precious No. 1,0.1~1.0g/L spend No. 2 precious, 0.1~1.0mg/L NAA, 15~30g/L sucrose+3.5~
6.0g/L agar, 50~100g/L bananas juices, 0.2~0.5g/L activated carbons, pH are 5.4~5.8.
In the seedling method for test tube propagation of above-mentioned wide leaf thickness lip orchid, the step 5 cultivate the time used be 20~
30 days.
In the seedling method for test tube propagation of above-mentioned wide leaf thickness lip orchid, the step 6 is specially：What step 5 obtained
Test tube seedling plant height is 3~5cm, and test tube seedling is shifted to the natural light lower refining seedling for being placed in greenhouse after 5~7 days, cleans the culture of root
It places to root system and whitens in the natural light in greenhouse after base, be 2 in volume ratio:2:1 bark, Lan Shi, mud stone mixed-matrix in
Seedling is cultivated up to seedling.
Beneficial effects of the present invention are as follows：
The present invention, into the seedling Propagations of Teat Tube Seedlings of line width leaf thickness lip orchid, it is blue to establish wide leaf thickness lip using plant tissue culture technique
Seedling Propagations of Teat Tube Seedlings technical system has the characteristics that seed germination rate is high, seedling is fast, seedling quality is good, at low cost.
The seed germination rate of the present invention reaches mainly by the component selection to culture medium used by axenic germination
100% germination rate.
The present invention further screens the culture medium of proliferation, its growth rate is allow to reach 3~5 times.
Further, the present invention further screens differentiation used medium, its differentiation rate is made to reach 89% or more；
Further, the present invention passes through the mixed-matrix to the preferred of the culture medium used in seedling culture and to transplanting
Control, makes its transplanting survival rate reach 93-100%.
Specific implementation mode
With reference to embodiment, technical scheme of the present invention is described in further detail, but do not constituted pair
Any restrictions of the present invention.
The seedling method for test tube propagation of the wide leaf thickness lip orchid of the present invention, including processing step below：
(1) explant acquires：It is explant to be developed to ripe and uncracked fruit from the wide leaf thickness lip orchid of field acquisition.
(2) axenic germination：Fruit described in step (1) is impregnated 1~5 minute in 75% ethyl alcohol, rinsed with sterile water 3
It is sterilized 10~30 minutes in 0.1% mercuric chloride solution after~5 times, 5~7 incision fruits of rinsed with sterile water, by its powdered embryo
It is inoculated into cultivate on axenic germination culture medium and can be obtained within 90~120 days protocorm, the axenic germination culture medium is：0.5
~2.0g/L spends precious No. 1 (HYPONeX 1)+0.2~1.0mg/L NAA+15~30g/L+3.5~6.0g/L of sucrose agar+50
~100ml/L coconut milk, pH are 5.4~5.8.
(3) Protocorm Multiplication：Protocorm obtained by step (2) is inoculated into Protocorm Multiplication culture medium and cultivates 30~45
It can be proliferated to obtain a large amount of protocorms, and growth coefficient is 3~5 times, and the Protocorm Multiplication culture medium is：Improvement
Knudson C+0.1~1.0mg/L 6-BA+0.01~0.3mg/L TDZ+15~30g/L+3.5~6.0g/L of sucrose agar+
50~100ml/L coconut milk+0.2~0.5g/L activated carbons, pH are 5.4~5.8.
(4) differentiation culture：Protocorms obtained by step (3) are inoculated into protocorm differentiation culture medium and cultivate 40~60
It can differentiate seedling, and the protocorm differentiation culture medium is：Improvement Knudson C+0.1~1.0mg/L NAA+15~
30g/L sucrose+3.5~6.0g/L agar+50~100ml/L coconut milk+0.2~0.5g/L activated carbons, pH are 5.4~5.8.
(5) strong seedling culture：The seedling that step (4) obtains is transferred to and cultivates 20~30 days and can be formed in strong seedling culture base
Test tube seedling, the strong seedling culture base are：It is No. 2 precious that 0.1~1.0g/L spends No. 1 (HYPONeX 1)+0.1~1.0g/L of treasured to spend
(HYPONeX 2)+0.1~1.0mg/L NAA+15~30g/L sucrose+3.5~6.0g/L+50~100g/L of agar bananas juices+
0.2~0.5g/L activated carbons, pH are 5.4~5.8.
(6) it transplants：The test tube seedling transfer that plant height obtained by step (5) is 3~5cm is placed in the natural light lower refining seedling 5 in greenhouse
After~7 days, the natural light after the culture medium of clean root in greenhouse, which is placed to root system, whitens, and is 2 in volume ratio:2:1 bark,
Lan Shi, mud stone mixed-matrix in cultivation seedling up to seedling.
The condition of culture of above-mentioned steps (2)~(5) is：Cultivation temperature be 25~28 DEG C, intensity of illumination be 1500~
2000lx, light application time are 12~16 hours/day.
In order to further be illustrated to the advantage of the present invention, the present invention provides following embodiment and is further discussed
Card.
Embodiment one
(1) explant acquires：It is explant to be developed to ripe and uncracked fruit from the wide leaf thickness lip orchid of field acquisition.
(2) axenic germination：Fruit described in step (1) is impregnated 2 minutes in 75% ethyl alcohol, rinsed with sterile water 3 times
It is sterilized 10 minutes in 0.1% mercuric chloride solution afterwards, cuts fruit after rinsed with sterile water 5 times, its powdered embryo is inoculated into sterile
It is cultivated 93 days on germination medium and can be obtained protocorm, pollution rate 5.9%, seed germination rate 100%.It is described sterile to sprout
Sending out culture medium is：1.0g/L spends precious No. 1 (HYPONeX 1)+0.5mg/L NAA+18g/L sucrose+4.0g/L agar+55ml/L coconut palm
Sub- juice, pH 5.6.
(3) Protocorm Multiplication：Protocorm obtained by step (2) is inoculated into Protocorm Multiplication culture medium and cultivates 30 days i.e.
It can be proliferated to obtain a large amount of protocorms, growth coefficient reaches 3.5 times, and the Protocorm Multiplication culture medium is：Improvement
Knudson C+0.5mg/L 6-BA+0.1mg/L TDZ+15g/L sucrose+3.5/L agar+80ml/L coconut milk+0.3g/L lives
Property charcoal, pH 5.6.
(4) differentiation culture：Protocorms obtained by step (3) are inoculated into protocorm differentiation culture medium and cultivate 40 days i.e.
Seedling can be differentiated, differentiation rate reaches 93% or more.The protocorm differentiation culture medium is：Improve Knudson C+0.3mg/
L NAA+15g/L sucrose+4.0g/L agar+50ml/L coconut milk+0.2g/L activated carbons, pH 5.6.
(5) strong seedling culture：The seedling that step (4) obtains is transferred to cultivate 20 days in strong seedling culture base and can obtain plant height
The test tube seedling of 3cm or more, the strong seedling culture base are：It is No. 2 precious that 0.5g/L spends No. 1 (HYPONeX 1)+0.5g/L of treasured to spend
(HYPONeX 2)+0.3mg/L NAA+20g/L sucrose+3.5g/L agar+50g/L bananas juice+0.2g/L activated carbons, pH are
5.6。
(6) it transplants：The test tube seedling transfer that plant height obtained by step (5) is 3~5cm is placed in the natural light lower refining seedling 5 in greenhouse
After it, the natural light after the culture medium of clean root in greenhouse, which is placed to root system, whitens, and is 2 in volume ratio:2:1 bark, orchid
Stone, mud stone mixed-matrix in cultivation seedling up to seedling, survival rate after transplanting 2 months is up to 96% or more.
The condition of culture of above-mentioned steps (2)~(5) is：Cultivation temperature is 25 DEG C, intensity of illumination 1500lx, light application time
For 12 hours/day.
Embodiment two
(1) explant acquires：It is explant to be developed to ripe and uncracked fruit from the wide leaf thickness lip orchid of field acquisition.
(2) axenic germination：Fruit described in step (1) is impregnated 1 minute in 75% ethyl alcohol, rinsed with sterile water 3 times
It is sterilized 10 minutes in 0.1% mercuric chloride solution afterwards, its powdered embryo is inoculated into sterile sprout by 5 incision fruits of rinsed with sterile water
It is cultivated 100 days on hair culture medium and can be obtained protocorm, pollution rate 10%, seed germination rate reaches 95% or more.Described
Axenic germination culture medium is：1.5g/L spend precious No. 1 (HYPONeX 1)+0.4mg/L NAA+30g/L sucrose+5.0g/L agar+
75ml/L coconut milk, pH 5.4.
(3) Protocorm Multiplication：Protocorm obtained by step (2) is inoculated into Protocorm Multiplication culture medium and cultivates 38 days i.e.
It can be proliferated to obtain a large amount of protocorms, growth coefficient reaches 4.2 times, and the Protocorm Multiplication culture medium is：Improvement
Knudson C+0.7mg/L 6-BA+0.2mg/L TDZ+20g/L sucrose+5.5g/L agar+50ml/L coconut milk+0.15g/L
Activated carbon, pH 5.4.
(4) differentiation culture：Protocorms obtained by step (3) are inoculated into protocorm differentiation culture medium and cultivate 43 days i.e.
Seedling can be differentiated, differentiation rate reaches 97% or more, and the protocorm differentiation culture medium is：Improve Knudson C+0.3mg/
L NAA+25g/L sucrose+4.3g/L agar+65ml/L coconut milk+0.35g/L activated carbons, pH 5.4.
(5) strong seedling culture：The seedling that step (4) obtains is transferred to cultivate 28 days in strong seedling culture base and can obtain plant height
The wide leaf thickness lip orchid test tube seedling of 4cm or more, the strong seedling culture base are：0.8g/L spends No. 1 (HYPONeX 1)+0.4g/L of treasured
Precious No. 2 (HYPONeX 2)+0.3mg/L NAA+30g/L sucrose+4.5g/L agar+75g/L bananas juice+0.3g/L activated carbons are spent,
PH is 5.4.
(6) it transplants：The test tube seedling transfer that plant height obtained by step (5) is 3~5cm is placed in the natural light lower refining seedling 6 in greenhouse
After it, the natural light after the culture medium of clean root in greenhouse, which is placed to root system, whitens, and is 2 in volume ratio:2:1 bark, orchid
Stone, mud stone mixed-matrix in cultivation seedling up to seedling, survival rate after transplanting 2 months is up to 93% or more.
The condition of culture of above-mentioned steps (2)~(5) is：Cultivation temperature is 26 DEG C, intensity of illumination 1800lx, light application time
For 14 hours/day.
Embodiment three
(1) explant acquires：It is explant to be developed to ripe and uncracked fruit from the wide leaf thickness lip orchid of field acquisition.
(2) axenic germination：Fruit described in step (1) is impregnated 5 minutes in 75% ethyl alcohol, rinsed with sterile water 5 times
It is sterilized 30 minutes in 0.1% mercuric chloride solution afterwards, its powdered embryo is inoculated into sterile sprout by 7 incision fruits of rinsed with sterile water
It is cultivated on hair culture medium and can be obtained within 110 days protocorm, pollution rate is less than 3%, and seed germination rate reaches 90% or more.Described
Axenic germination culture medium is：2.0g/L spend precious No. 1 (HYPONeX 1)+1.0mg/L NAA+30g/L sucrose+6.0g/L agar+
100ml/L coconut milk, pH 5.8.
(3) Protocorm Multiplication：Protocorm obtained by step (2) is inoculated into Protocorm Multiplication culture medium and cultivates 42 days i.e.
It can be proliferated to obtain a large amount of protocorms, growth coefficient is 4.6 times, and the Protocorm Multiplication culture medium is：Improve Knudson
C+1.0mg/L 6-BA+0.3mg/L TDZ+30g/L sucrose+6.0g/L agar+100ml/L coconut milk+0.5g/L activated carbons, pH
It is 5.8.
(4) differentiation culture：Protocorms obtained by step (3) are inoculated into protocorm differentiation culture medium and cultivate 40 days i.e.
Seedling can be differentiated, differentiation rate reaches 89%.The protocorm differentiation culture medium is：Improve Knudson C+1.0mg/L
NAA+30g/L sucrose+6.0g/L agar+100ml/L coconut milk+0.5g/L activated carbons, pH 5.8.
(5) strong seedling culture：The seedling that step (4) obtains is transferred to cultivate 25 days in strong seedling culture base and can obtain plant height
The wide leaf thickness lip orchid test tube seedling of about 5cm, the strong seedling culture base are：0.1g/L spends precious No. 1 (HYPONeX 1)+1.0g/L flowers
Precious No. 2 (HYPONeX 2)+1.0mg/L NAA+30g/L sucrose+6.0g/L agar+100g/L bananas juice+0.5g/L activated carbons,
PH is 5.8.
(6) it transplants：The test tube seedling transfer that plant height obtained by step (5) is 3~5cm is placed in the natural light lower refining seedling 7 in greenhouse
After it, the natural light after the culture medium of clean root in greenhouse, which is placed to root system, whitens, and is 2 in volume ratio:2:1 bark, orchid
Stone, mud stone mixed-matrix in cultivation seedling up to seedling.Survival rate after transplanting 2 months reaches 100%.
The condition of culture of above-mentioned steps (2)~(5) is：Cultivation temperature is 28 DEG C, intensity of illumination 2000lx, light application time
For 15 hours/day.
It is above-described be only presently preferred embodiments of the present invention, it is all within the scope of the spirit and principles in the present invention made by appoint
What modifications, equivalent substitutions and improvements etc., should all be included in the protection scope of the present invention.
Claims (7)
1. a kind of seedling method for test tube propagation of width leaf thickness lip orchid, it is characterised in that：Include the following steps：
Step 1：The wide leaf thickness lip orchid of acquisition is developed to ripe and uncracked fruit as explant；
Step 2：Powdered embryo in explant is subjected to axenic germination and obtains protocorm, is trained used by the axenic germination
Support base constituent content be：0.5~2.0g/L spends No. 1 precious, 0.2~1.0mg/L NAA, 15~30g/L sucrose, 3.5~6.0g/
L agar, 50~100ml/L coconut milk, pH are 5.4~5.8；
Step 3：Protocorm Multiplication is obtained into protocorms；Wherein, the group of the culture medium used in the proliferation is divided into：Improvement
Knudson C, 0.1~1.0mg/L 6-BA, 0.01~0.3mg/L TDZ, 15~30g/L sucrose, 3.5~6.0g/L agar,
50~100ml/L coconut milk, 0.2~0.5g/L activated carbons, pH are 5.4~5.8；
Step 4：Protocorms differentiation culture is obtained into seedling, the group for breaking up culture medium used is divided into：Improvement Knudson C,
0.1~1.0mg/L NAA, 15~30g/L sucrose, 3.5~6.0g/L agar, 50~100ml/L coconut milk, 0.2~0.5g/L
Activated carbon, pH are 5.4~5.8；
Step 5：Seedling culture is obtained into test tube seedling, the group of culture medium used is divided into：0.1~1.0g/L spend precious No. 1,0.1~
1.0g/L spends No. 2 precious, 0.1~1.0mg/L NAA, 15~30g/L+3.5~6.0g/L of sucrose agar, 50~100g/L bananas
Juice, 0.2~0.5g/L activated carbons, pH are 5.4~5.8；
Step 6：By test tube transplantation of seedlings up to seedling.
2. the seedling method for test tube propagation of width leaf thickness lip orchid according to claim 1, which is characterized in that the step 2
In, the acquisition methods of the powdered embryo in explant are：Explant is impregnated 1~5 minute in ethyl alcohol, rinsed with sterile water 3
It is sterilized 10~30 minutes in mercuric chloride solution after~5 times, 5~7 incision fruits of rinsed with sterile water obtain powdered embryo.
3. the seedling method for test tube propagation of width leaf thickness lip orchid according to claim 1, which is characterized in that the step 2
In, incubation time is 90~120 days to powdered embryo in the medium.
4. the seedling method for test tube propagation of width leaf thickness lip orchid according to claim 1, which is characterized in that the step 3
In incubation time be 30~45 days, growth coefficient be 3~5 times.
5. the seedling method for test tube propagation of width leaf thickness lip orchid according to claim 1, which is characterized in that the step 4
In, the differentiation culture time used is 40~60 days.
6. the seedling method for test tube propagation of width leaf thickness lip orchid according to claim 1, which is characterized in that the step 5
The culture time used is 20~30 days.
7. the seedling method for test tube propagation of width leaf thickness lip orchid according to claim 1, which is characterized in that the step 6
Specially：The test tube seedling plant height that step 5 obtains is 3~5cm, and test tube seedling transfer is placed in the natural light lower refining seedling 5~7 days in greenhouse
Afterwards, the natural light after the culture medium of clean root in greenhouse, which is placed to root system, whitens, and is 2 in volume ratio:2:1 bark, Lan Shi,
Seedling is cultivated in the mixed-matrix of mud stone up to seedling.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610935238.2A CN106538384B (en) | 2016-11-01 | 2016-11-01 | A kind of seedling method for test tube propagation of width leaf thickness lip orchid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610935238.2A CN106538384B (en) | 2016-11-01 | 2016-11-01 | A kind of seedling method for test tube propagation of width leaf thickness lip orchid |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106538384A CN106538384A (en) | 2017-03-29 |
| CN106538384B true CN106538384B (en) | 2018-08-24 |
Family
ID=58392323
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610935238.2A Expired - Fee Related CN106538384B (en) | 2016-11-01 | 2016-11-01 | A kind of seedling method for test tube propagation of width leaf thickness lip orchid |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106538384B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104082123A (en) * | 2014-06-28 | 2014-10-08 | 玉林师范学院 | Cultivation method of tetraploid Anoectochilus roxburghii |
| CN104770294A (en) * | 2015-04-02 | 2015-07-15 | 河南省农业科学院 | Breeding method using protocorm based on germinated phalaenopsis seeds as receptor |
| CN105359983A (en) * | 2015-12-24 | 2016-03-02 | 陕西师范大学 | Dendrobium officinale Kimura et Migo protocorm direct-seeding method |
-
2016
- 2016-11-01 CN CN201610935238.2A patent/CN106538384B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104082123A (en) * | 2014-06-28 | 2014-10-08 | 玉林师范学院 | Cultivation method of tetraploid Anoectochilus roxburghii |
| CN104770294A (en) * | 2015-04-02 | 2015-07-15 | 河南省农业科学院 | Breeding method using protocorm based on germinated phalaenopsis seeds as receptor |
| CN105359983A (en) * | 2015-12-24 | 2016-03-02 | 陕西师范大学 | Dendrobium officinale Kimura et Migo protocorm direct-seeding method |
Non-Patent Citations (1)
| Title |
|---|
| 金线莲的结实特性和无菌播种培养;何荆洲等;《江苏农业科学》;20141231;第42卷(第9期);第214-217页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106538384A (en) | 2017-03-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104067821B (en) | A kind of preparation method of Sweetpotato Viruses Elimination seedling | |
| CN101983554B (en) | Reproduction method of Chinese cymbidium seedlings | |
| CN107047320B (en) | A kind of bigflower centranthera root method for tissue culture | |
| CN101926259B (en) | Orchid germchit propagating method | |
| CN103931497B (en) | A kind of method improving dragon fruit plantlet in vitro planting percent | |
| CN105766654B (en) | A kind of Nanchuan jackfruit method for tissue culture | |
| CN104041412A (en) | Rapid propagation method for tissue culture of Guizhou hemiboea cavaleriei | |
| CN104106468A (en) | Tissue culture and rapid propagation method of radix fici simplicissimae | |
| CN106472306B (en) | One kind begins to flourish stem of noble dendrobium high quality seedling asexual clonal rapid propagation method | |
| CN105475129A (en) | Tissue-culture rapid propagation method for arundina graminifolia | |
| CN103039363B (en) | Rooting medium for tissue culture seedling propagation of camellia oleifera abel and propagation method thereof | |
| CN105104200B (en) | A kind of quick breeding method for tissue culture of Sinia rhodoleuca | |
| CN106577280A (en) | Rapid propagation aseptic seedling by means of tender stem segments and blades of merrillanthus hainanensis | |
| CN103039362A (en) | Subculture medium for tissue culture seedling propagation of camellia oleifera abel and propagation method thereof | |
| CN110122335A (en) | The abductive approach of Xu Xiang Kiwifruit Tissue Culture seedling salt-tolerant mutant | |
| CN105145358A (en) | Tissue culture and rapid propagation method for common fibraurea stem | |
| CN112106664B (en) | Sterile germination and rapid propagation method for michelia spectabilis seeds | |
| CN105766641B (en) | A kind of method of yam stem section Vitro Quick Reproduction | |
| CN106538384B (en) | A kind of seedling method for test tube propagation of width leaf thickness lip orchid | |
| CN108124770A (en) | A kind of wonga-wonga stem of noble dendrobium high quality seedling asexual clonal rapid propagation method | |
| CN108112479A (en) | A kind of stem section of papaya sprout Bud Differentiation vacantly plants leaf promoting root growth method | |
| CN104186324A (en) | Method for inducing mature zygotic embryo somatic cell embryos of pinus sylvestris var mongolica | |
| CN101012448A (en) | Method of screening leymus chinensis salt-tolerant somatic mutation | |
| CN103053429B (en) | Method for regenerating semen pharbitidis in vitro embryonic axis plant | |
| CN105794650A (en) | Method for preserving minimum population Guangxi bilberry offspring by means of immature seeds |
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: 20180824 Termination date: 20191101 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |