CN105660415A - In-vitro preservation method of drynaria roosii nakaike - Google Patents
In-vitro preservation method of drynaria roosii nakaike Download PDFInfo
- Publication number
- CN105660415A CN105660415A CN201610108548.7A CN201610108548A CN105660415A CN 105660415 A CN105660415 A CN 105660415A CN 201610108548 A CN201610108548 A CN 201610108548A CN 105660415 A CN105660415 A CN 105660415A
- Authority
- CN
- China
- Prior art keywords
- culture medium
- mongolian oak
- pteridii latiusculi
- herba pteridii
- days
- 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
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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
-
- 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)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Developmental Biology & Embryology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Cultivation Of Plants (AREA)
Abstract
The invention discloses an in-vitro preservation method of drynaria roosii nakaike, which comprises: obtaining tissue culture two-step adult seedlings of drynaria roosii nakaike; then obtaining test-tube plantlets each of which has 4 to 6 leaves by utilizing the tissue culture two-step adult seedlings; inoculating the test-tube plantlets to a preservation culture medium to carry out in-vitro preservation. The preservation culture medium adopted by the in-vitro preservation method comprises 1/2 MS, 0.3% of AC, 4.5% of sucrose and 0.4% of agar. The culture medium has a pH value of 5.8, a temperature of 23 plus and minus 1 DEG C, illumination intensity of 1,400 to 1,600lx, and illumination time of 8 to 10 hours per day. By utilizing the method disclosed by the invention, a great number of high-quality seedlings of drynaria roosii nakaike can be provided in short time, meanwhile, a step of preparing the culture medium and a spinner bottle is reduced, and a great quantity of manpower and material resources are saved, and thus, the method is simple and easy to operate, is reduced in production cost, and effectively solves the problems of preservation of genetic resources and large-scale seedling culture of drynaria roosii nakaike.
Description
Technical field
The invention belongs to technical field of tissue culture, in particular it relates to the in-vitro conservation method of Mongolian oak Herba pteridii latiusculi.
Background technology
Mongolian oak Herba pteridii latiusculi DrynariaroosiiNakaike is Plants of Polypodiaceae, sprawl growth, or grows nonparasitically upon another plant on trunk, wall or on cliff, helical form is climbed up by holding on to, and is born in the woods of mountain region on trunk or on rock, height above sea level 500-700 rice. Mongolian oak Herba pteridii latiusculi adopts sporogenesis, and rhizome is used as medicine, and has the kidney invigorating bone strengthening, the continuous effect hindering pain relieving. For lumbago due to renal deficiency, chronic diarrhea of suffering from a deficiency of the kidney, Hiccough and deaf, odontoseisis, fall wink frustrate, injured bone; External treatment alopecia areata, vitiligo. Mongolian oak Herba pteridii latiusculi produces Jiangsu, Fujian, Taiwan, Guangxi etc. and economizes. It is born in the woods of mountain region on trunk or on rock, height above sea level 500-700 rice. Owing to growth in the living standard people are also more and more higher to the pursuit of health idea, Mongolian oak Herba pteridii latiusculi is developed to numerous medical material, on market, various one-tenth capsules, bone pine recovering particles are almost all with Mongolian oak Herba pteridii latiusculi medical material Rhizoma Drynariae for primary raw material, the also medicine such as stomach medicine and Anti-hair loss of recent development. About in JIUYUE, 2015 price 20 yuan/jin. Due to Mongolian oak Herba pteridii latiusculi poor growth, excavating serious, existing market is rare, and wild Mongolian oak Herba pteridii latiusculi resource falls sharply, and the research cultivated and breed is still less, manually introduces a fine variety breeding imperative.
Summary of the invention
For the problem that presently, there are, the present invention provides the in-vitro conservation method of a kind of Mongolian oak Herba pteridii latiusculi, utilize tissue culture technique that it is carried out group training two step seedling-growing rapid breedings, by adopting suitable Storaged media that it is carried out in vitro conservation, it is possible not only to effectively save Mongolian oak Herba pteridii latiusculi species in imminent danger, make Renewable resource, it is also possible to provide the technology of excellent consistent seedling and indoor in vitro conservation to provide technical support for the plantation of Mongolian oak Herba pteridii latiusculi.
For achieving the above object, the technical scheme is that
(1) spore germination is cultivated: take Mongolian oak Herba pteridii latiusculi maturation dry as outer implant, 75% ethanol soaks 30 ~ 45s, aseptic water washing 2 ~ 3 times, put into the 0.1% mercuric chloride 4min adding 2 tweens, use aseptic water washing 2 ~ 3 times again, again put into the 0.1% mercuric chloride 4min adding 2 tweens, finally it is seeded on inducing spore germination medium with being cut into 0.5 ~ 1.0cm length after aseptic water washing 2 ~ 3 times, cultivate 10 ~ 14 days, the visible peak green speckle of spore germination, and become changeable green gradually, a large amount of original foliage occurred in 30 ~ 40 days, a large amount of sporinite occurred in 60 ~ 70 days, it is emerald green for becoming a small bundle of straw, etc. for silkworms to spin cocoons on,
(2) sporinite breeding root culture: cultivated through 80 ~ 90 days, sporinite grows up to and has 2 needles, high about 2cm sprout, sprout is taken off and is inoculated on breeding root media and cultivates, 30 ~ 45 days, aseptic seedling breeding coefficient 4 ~ 7, plant height 4.0 ~ 6.0, mean elements more than 4, average more than root length 4cm;
(3) test tube Seedling in vitro conservation: by the test tube Seedling with 4 ~ 6 leaves of acquisition of being taken root by sporinite breeding, be inoculated into Storaged media and carry out in vitro conservation.
Germination medium described in step (1) is: MS+TDZ1.0mg/l+NAA0.5mg/l+AC0.5%+ sucrose 2.5%+ agar 0.4%; The pH value of described culture medium is 5.8, and cultivation temperature is 24 ~ 28 ° of C, intensity of illumination 1400 ~ 2000lx, and light application time is 12 ~ 14 hours/day.
Breeding root media described in step (2) is: MS+IAA1.0mg/l+6-BA0.6mg/l+AC0.5%+ sucrose 3.5%+ agar 0.4%; The pH value of described culture medium is 5.8, and temperature is 22 ~ 24 ° of C, intensity of illumination 2600 ~ 2800lx, and light application time is 12 ~ 14 hours/day.
Storaged media described in step (3) is 1/2MS+CCC1.0mg/l+AC1.0%+ sucrose 4.5%+ agar 0.4%; The pH value of described culture medium is 5.8, and temperature is 23 ± 1 ° of C, intensity of illumination 1200 ~ 1600lx, and light application time is 6 ~ 8 hours/day.
Beneficial effects of the present invention:
Though the present invention and patent 200410098498.6 are all Mongolian oak Herba pteridii latiusculi sporogenesis, but the breeding in Mongolian oak Herba pteridii latiusculi tissue-culturing rapid propagation process and taking root is carried out by the present invention simultaneously, decrease a step, same expanding propagation effect can be reached, therefore the present invention is adopted will to save manpower, material resources and time, simultaneously, the present invention has broken the blank to the research of Mongolian oak Herba pteridii latiusculi in vitro conservation, the method utilizing the present invention can make germ plasm resource obtain long-term preservation, and the seedling obtained is healthy and strong, survival rate is high, the high quality seedling of a large amount of Mongolian oak Herba pteridii latiusculi can be provided in a short time, efficiently solve Mongolian oak Cyclosorus Germ-plasma resources protection and scale breeding problem.
Accompanying drawing explanation
Fig. 1 represents the light intensity of the present invention impact on Mongolian oak Herba pteridii latiusculi in vitro conservation.
Wherein, abscissa represents intensity of illumination (lx), and vertical coordinate represents preservation natural law (d).
Detailed description of the invention
The experimental technique used in following embodiment if no special instructions, is conventional method.
Material used in following embodiment, reagent etc., if no special instructions, all commercially obtain.
A kind of Mongolian oak Cyclosorus in-vitro conservation method, is realized by the following method:
One, Mongolian oak Herba pteridii latiusculi obtains culture through Fast-propagation:
1, material to be tested
Mongolian oak Herba pteridii latiusculi DrynariaroosiiNakaike spore.
2, spore germination is cultivated:
Take Mongolian oak Herba pteridii latiusculi maturation dry as outer implant, 75% ethanol soaks 30 ~ 45s, aseptic water washing 2 ~ 3 times, put into the 0.1% mercuric chloride 4min adding 2 tweens, use aseptic water washing 2 ~ 3 times again, again put into the 0.1% mercuric chloride 4min adding 2 tweens, finally it is seeded on MS+TDZ1.0mg/l+NAA0.5mg/l+AC0.5%+ sucrose 2.5%+ agar 0.4% germination medium with being cut into 0.5 ~ 1.0cm length after aseptic water washing 2 ~ 3 times, the pH value of described culture medium is 5.8, cultivation temperature is 24 ~ 28 ° of C, intensity of illumination 1400 ~ 2000lx, light application time is 12 ~ 14 hours/day, cultivate 10 ~ 14 days, the visible peak green speckle of spore germination, and become changeable green gradually, a large amount of original foliage occurred in 30 ~ 40 days, a large amount of sporinite occurred in 60 ~ 70 days, it is emerald green for becoming a small bundle of straw, etc. for silkworms to spin cocoons on,
3, sporinite breeding root culture
Cultivating through 80 ~ 90 days, sporinite grows up to and has 2 needles, high about 2cm sprout, sprout is taken off and is inoculated into breeding MS+IAA1.0mg/l+6-BA0.6mg/l+AC1.0%+ sucrose 3.5%+ agar 0.4%; The pH value of described culture medium is 5.8, and temperature is 22 ~ 25 ° of C, intensity of illumination 2600 ~ 2800lx, and light application time is 12 ~ 14 hours/day, 30 ~ 45 days, aseptic seedling breeding coefficient 4 ~ 7, plant height 4.0 ~ 6.0, mean elements more than 4, average more than root length 4cm.
5, test tube Seedling in vitro conservation
Select above-mentioned have 4 ~ 6 leaves, more than 4 radical Mongolian oak Herba pteridii latiusculi test tube Seedlings are that test material carries out in vitro conservation.
Below it is carried out the screening of type of culture medium, temperature, intensity of illumination, growth inhibitor, preserve natural law and all add up for 60% with test tube Seedling survival rate.
(1), the culture medium impact on Mongolian oak Herba pteridii latiusculi in vitro conservation
Test material is inoculated in EXPERIMENTAL DESIGN MS, tri-kinds of culture medium of 1/2MS, 1/4MS, inoculates 10 bottles, every bottle of 5 strain simple buds; Containing the agar of 35g/L sucrose and 4g/L in culture medium, the pH value of culture medium is 5.8, cultivation temperature 25 ± 1 DEG C, light intensity 2000lx, illumination 8h/d; It is shown that Mongolian oak Herba pteridii latiusculi holding time in 1/2MS culture medium is the longest, being 168 days, concrete outcome is in Table 1.
The impact on the Mongolian oak Herba pteridii latiusculi in vitro conservation time of table 1 culture medium
| Culture medium | Holding time (d) | Upgrowth situation |
| MS | 149 | Plant is light green, well-grown, starts death after 4 months |
| 1/2MS | 168 | Plant is light green, well-grown, starts death after 5 months, |
| 1/4MS | 157 | Bud strain poor growth, after 3 months, material hops to it withered dying, |
(2), the temperature impact on Mongolian oak Herba pteridii latiusculi in vitro conservation
Test material accessing 1/2MS culture medium and is respectively placed in temperature 17 DEG C, 20 DEG C, the illumination box of 23 DEG C, 26 DEG C is cultivated, and inoculates 10 bottles, every bottle of 5 strain simple buds; Agar containing 30g/L sucrose and 4.85g/L in culture medium, the pH value of culture medium is 5.8, light intensity 2000lx, illumination 7h/d, it is shown that Mongolian oak Herba pteridii latiusculi temperature be in 23 DEG C of incubators the holding time the longest, be 196 days; It is specifically shown in table 2.
The impact on the Mongolian oak Herba pteridii latiusculi in vitro conservation time of table 2 temperature
(3), the intensity of illumination impact on Mongolian oak Herba pteridii latiusculi in vitro conservation
Test material is inoculated into 1/2MS culture medium, design 0,1200lx, 1400lx, five kinds of intensities of illumination of 1600lx, 1800lx, and every kind of light intensity inoculates 10 bottles, every bottle of 5 strain simple buds; Containing the agar of 35g/L sucrose and 4.5g/L in culture medium, the pH value of culture medium is 5.8, cultivation temperature 23 ± 1 DEG C, illumination 7h/d; It is shown that Mongolian oak Herba pteridii latiusculi holding time when light intensity is 1600lx is the longest, being 251 days, concrete comparative result is shown in Fig. 1, and abscissa is for preserving natural law, and vertical coordinate is intensity of illumination.
(4), the growth inhibitor impact on Mongolian oak Herba pteridii latiusculi in vitro conservation
Test material is inoculated into interpolation following variable concentrations CCC, PPP3331/2MS culture medium on, inoculate 10 bottles, every bottle of 5 strain simple buds; Containing the agar of 30g/L sucrose and 4.5g/L in culture medium, the pH value of culture medium is 5.8, cultivation temperature 21 ± 1 DEG C, light intensity 1600lx, illumination 7h/d, it is shown that Mongolian oak Herba pteridii latiusculi is 1.0mg/L in CCC concentration is that the holding time is the longest, is 274 days; Concrete condition is in Table 3.
The impact on the Mongolian oak Herba pteridii latiusculi holding time of table 3 hormone
| Hormone | Concentration (mg/L) | Holding time (d) | Upgrowth situation |
| CCC | 0.5 | 229 | Plant is light green, well-grown, and 5 months start death |
| CCC | 1.0 | 274 | Plant is light green, well-grown, and 6 months start death |
| CCC | 1.5 | 261 | Plant is light green, well-grown, and 5 months start death |
| PP333 | 0.5 | 217 | Plant is light green, well-grown, and 6 months start death |
| PP333 | 1.0 | 241 | Plant is light green, well-grown, and 6 months start death |
| PP333 | 1.5 | 254 | Plant is light green, well-grown, and 6 months start death |
By above-mentioned experiment, it is determined that Mongolian oak Herba pteridii latiusculi the best in vitro conservation culture medium is 1/2MS+CCC1.0mg/l+AC1.0%+ sucrose 3.5%+ agar 0.4%;The pH value of described culture medium is 5.8, and temperature is 23 ± 1 ° of C, intensity of illumination 1400 ~ 1600lx, and light application time is 6 ~ 8 hours/day. Preserving natural law is 274 days.
5, upgrowth situation is investigated
The growth recovery situation of the Mongolian oak Herba pteridii latiusculi test tube Seedling of in vitro conservation is investigated: is cut by the Mongolian oak Herba pteridii latiusculi test tube Seedling simple bud of robust growth and is inoculated on best Storaged media after preservation 274d, the Mongolian oak Herba pteridii latiusculi test tube Seedling of survival is inoculated on breeding root media MS+IAA1.0mg/l+6-BA0.6mg/l+AC0.5%, add on the sucrose of 3% and the agar culture medium of 0.45%, every 30d subculture 1 time, after subculture 3 times, the recovery situation of Mongolian oak Herba pteridii latiusculi test tube Seedling is investigated, in Table 4 with plant height, rate of growth, breeding rate, individual plant rooting rate for index. It can be seen that the test tube Seedling recovery situation after 274d in vitro conservation is good from contrast.
The test tube Seedling recovered after table 4 in vitro conservation and the comparison preserving front test tube Seedling
| Material | Average life rate/% | Breeding coefficient/times | Rooting rate/% | Plant height/cm |
| Normally | 6.64±0.11 | 7.1±0.50 | 97.1±0.51 | 7.01±0.13 |
| Preserve | 6.25±0.24 | 6.7±0.32 | 96.3±0.32 | 6.67±0.12 |
Material used in the present invention annotates respectively as TDZ (phenylurea), 6-BA(6-benayl aminopurine), NAA(naphthalene acetic acid), IAA (heteroauxing), AC (activated carbon), CCC(chlorocholine chloride), PPP333(paclobutrazol).
Claims (4)
1. the in-vitro conservation method of a Mongolian oak Herba pteridii latiusculi, it is characterised in that comprise the steps:
(1) spore germination is cultivated: take Mongolian oak Herba pteridii latiusculi maturation dry as outer implant, 75% ethanol soaks 30 ~ 45s, aseptic water washing 2 ~ 3 times, put into the 0.1% mercuric chloride 4min adding 2 tweens, use aseptic water washing 2 ~ 3 times again, again put into the 0.1% mercuric chloride 4min adding 2 tweens, finally it is seeded on inducing spore germination medium with being cut into 0.5 ~ 1.0cm length after aseptic water washing 2 ~ 3 times, cultivate 10 ~ 14 days, the visible peak green speckle of spore germination, and become changeable green gradually, a large amount of original foliage occurred in 30 ~ 40 days, a large amount of sporinite occurred in 60 ~ 70 days, it is emerald green for becoming a small bundle of straw, etc. for silkworms to spin cocoons on,
(2) sporinite breeding root culture: cultivated through 80 ~ 90 days, sporinite grows up to and has 2 needles, high about 2cm sprout, sprout is taken off and is inoculated on breeding root media and cultivates, 30 ~ 45 days, aseptic seedling breeding coefficient 4 ~ 7, plant height 4.0 ~ 6.0, mean elements more than 4, average more than root length 4cm;
(3) test tube Seedling in vitro conservation: by the test tube Seedling with 4 ~ 6 leaves of acquisition of being taken root by sporinite breeding, be inoculated into Storaged media and carry out in vitro conservation.
2. the in-vitro conservation method of Mongolian oak Herba pteridii latiusculi as claimed in claim 1, it is characterised in that the germination medium described in step (1) is: MS+TDZ1.0mg/l+NAA0.5mg/l+AC0.5%+ sucrose 2.5%+ agar 0.4%; The pH value of described culture medium is 5.8, and cultivation temperature is 24 ~ 28 ° of C, intensity of illumination 1400 ~ 2000lx, and light application time is 12 ~ 14 hours/day.
3. the in-vitro conservation method of Mongolian oak Herba pteridii latiusculi as claimed in claim 1, it is characterised in that the breeding root media described in step (2) is: MS+IAA1.0mg/l+6-BA0.6mg/l+AC0.5%+ sucrose 3.5%+ agar 0.4%; The pH value of described culture medium is 5.8, and temperature is 22 ~ 25 ° of C, intensity of illumination 2600 ~ 2800lx, and light application time is 12 ~ 14 hours/day.
4. the in-vitro conservation method of Mongolian oak Herba pteridii latiusculi as claimed in claim 1, it is characterised in that the Storaged media described in step (3) is 1/2MS+CCC1.0mg/l+AC1.0%+ sucrose 4.5%+ agar 0.4%; The pH value of described culture medium is 5.8, and temperature is 23 ± 1 ° of C, intensity of illumination 1200 ~ 1600lx, and light application time is 6 ~ 8 hours/day.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610108548.7A CN105660415B (en) | 2016-02-29 | 2016-02-29 | A kind of in-vitro conservation method of Mongolian oak fern |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610108548.7A CN105660415B (en) | 2016-02-29 | 2016-02-29 | A kind of in-vitro conservation method of Mongolian oak fern |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105660415A true CN105660415A (en) | 2016-06-15 |
| CN105660415B CN105660415B (en) | 2017-09-19 |
Family
ID=56306108
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610108548.7A Expired - Fee Related CN105660415B (en) | 2016-02-29 | 2016-02-29 | A kind of in-vitro conservation method of Mongolian oak fern |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105660415B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106613809A (en) * | 2016-09-18 | 2017-05-10 | 大顺国际花卉股份有限公司 | All-year cultivation method for drynaria fortunei seedlings |
| CN109691372A (en) * | 2019-03-08 | 2019-04-30 | 中国科学院西双版纳热带植物园 | A kind of Mongolian oak fern artificial fecundation method |
| CN112293253A (en) * | 2020-10-28 | 2021-02-02 | 中国科学院昆明植物研究所 | Isolated preservation culture medium and isolated preservation method for Adiantum furiosaeanum |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1788549A (en) * | 2004-12-13 | 2006-06-21 | 中国科学院植物研究所 | Drynaria fortunei breeding method |
| CN103430853A (en) * | 2013-09-17 | 2013-12-11 | 南京通泽农业科技有限公司 | Rapid propagation method of Drynaria roosii |
-
2016
- 2016-02-29 CN CN201610108548.7A patent/CN105660415B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1788549A (en) * | 2004-12-13 | 2006-06-21 | 中国科学院植物研究所 | Drynaria fortunei breeding method |
| CN103430853A (en) * | 2013-09-17 | 2013-12-11 | 南京通泽农业科技有限公司 | Rapid propagation method of Drynaria roosii |
Non-Patent Citations (4)
| Title |
|---|
| 李娟等: "槲蕨愈伤组织的诱导研究", 《现代中药研究与实践》 * |
| 涂艺声: "《经济植物大规模快速繁殖技术》", 31 March 2009 * |
| 石雷等: "水龙骨科植物的引种及种质资源保存", 《植物资源与环境》 * |
| 韦莹等: "植物生长调节剂对槲蕨离体培养的效应", 《湖北农业科学院》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106613809A (en) * | 2016-09-18 | 2017-05-10 | 大顺国际花卉股份有限公司 | All-year cultivation method for drynaria fortunei seedlings |
| CN109691372A (en) * | 2019-03-08 | 2019-04-30 | 中国科学院西双版纳热带植物园 | A kind of Mongolian oak fern artificial fecundation method |
| CN112293253A (en) * | 2020-10-28 | 2021-02-02 | 中国科学院昆明植物研究所 | Isolated preservation culture medium and isolated preservation method for Adiantum furiosaeanum |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105660415B (en) | 2017-09-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101258835B (en) | Fast reproducing method for high quality seedling of dendrobium officinale | |
| CN102144553B (en) | Method for rapidly propagating Paris polyphylla Smith | |
| CN104429965B (en) | The method that high eyebrow Herba Anoectochili roxburghii seed is bred without hormone quickly tissue culture | |
| CN103734014B (en) | A kind of quick breeding method for tissue culture of anisetree bark | |
| CN104106468B (en) | The quick breeding method for tissue culture of a kind of radix fici simplicissimae | |
| CN104041412A (en) | Rapid propagation method for tissue culture of Guizhou hemiboea cavaleriei | |
| CN105475137A (en) | Method for tissue culture taking lotus reproduction and rooting into account | |
| CN106538387B (en) | A kind of method for tissue culture of Ku Zhi | |
| CN105660415A (en) | In-vitro preservation method of drynaria roosii nakaike | |
| CN106577291A (en) | Method for callus efficient induction seedling development of stemona sessilifolia | |
| CN104285792B (en) | A kind of tissue rapid propagation method of Alnus formosana Plantation | |
| CN103548695B (en) | A kind of meadowrueleaf corydalis root quick breeding method for tissue culture | |
| CN104813932A (en) | In vitro preservation method for Dendrobium officinale | |
| CN105210884A (en) | Bletilla seed asepsis sprouting equilibrated Medium | |
| CN102499082A (en) | Test tube breeding method of lilium oriental hybrid seed ball | |
| CN104686342A (en) | Asexual and rapid propagation technology for styrax tonkinensis | |
| CN105746347B (en) | A kind of in-vitro conservation method of meadowrueleaf corydalis root | |
| CN104756863B (en) | A kind of in-vitro conservation method of south China half capsule lettuce tongue | |
| CN102948370A (en) | Rapid propagation method of nothapodytes pittosporoides | |
| CN113317205A (en) | Efficient artificial seedling culture method for culturing amomum villosum by utilizing basal stem cluster buds and node propagation | |
| CN112616663A (en) | Method for greatly shortening planting period of lilium davidii var davidii and rapidly propagating seedlings | |
| CN104904592B (en) | A kind of in-vitro conservation method of Hemiboea lungzhouensis W. T Wang ex Z. Y. Li. | |
| CN104782501B (en) | May melon vine seedling cultural method | |
| CN116584389B (en) | Tissue culture and rapid propagation method of cress | |
| CN116098063B (en) | Rapid propagation method and application of test-tube seedlings of lycoris radiata leaf sheath-induced test-tube bulblet |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into 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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170919 Termination date: 20180229 |