CN105613296A - Tissue culture inoculating method for dendrobium officinale - Google Patents
Tissue culture inoculating method for dendrobium officinale Download PDFInfo
- Publication number
- CN105613296A CN105613296A CN201610046856.1A CN201610046856A CN105613296A CN 105613296 A CN105613296 A CN 105613296A CN 201610046856 A CN201610046856 A CN 201610046856A CN 105613296 A CN105613296 A CN 105613296A
- Authority
- CN
- China
- Prior art keywords
- embryo
- tissue culture
- dendrobium officinale
- inoculation
- root media
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—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)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a tissue culture inoculating method for dendrobium officinale. The tissue culture inoculating method for the dendrobium officinale comprises the following steps: selection of dendrobium officinale pods, sowing of seed embryos, preparation of sterile seed embryo mixed liquid, preparation of a germinating and rooting culture medium, and inoculation and culture of the seed embryos. The method disclosed by the invention is a simple and effective tissue culture inoculating method. According to the method, the germinating and rooting culture medium is inoculated to the cultured seed embryos through once induction, so that repeated culture rotation is not required; moreover, the germinating and rooting culture medium prepared in the method disclosed by the invention is mainly based on a natural antibacterial additive, and does not have harmful chemical component additive, so that the problems that the debugging time is long, the workload is long, the pollution rate is high, and the stability of produced seed seedlings is poor in conventional artificial one-by-one inoculation can be effectively solved; the pollution rate in the tissue culture process of the dendrobium officinale can be greatly reduced, and the tissue culture cost of the dendrobium officinale can be reduced; the tissue culture inoculating efficiency and the transplanting survival rate of the dendrobium officinale are greatly improved; the production efficiency is 3 to 4 times that of the conventional method; the transplanting survival rate can reach more than or equal to 92 to 93 percent.
Description
Technical field
The invention belongs to plant biotechnology field, be specifically related to a kind of candidum tissue culturing inoculation method.
Background technology
Herba Dendrobii (Dendrobiumofficinale) for the orchid family Dendrobium herbaceos perennial, dendrobium officinale is mainly distributed on the ground such as Zhejiang, Anhui, Jiangxi, Guangxi. The seed structure uniqueness of Herba Dendrobii, without endosperm, natural breed rate is extremely low, and the madness of wild resource is excavated by people in addition, and dendrobium officinale resource is endangered. Along with the development of modern biotechnology tissue culture technology, the artificial breeding technology of present seedlings of Dendrobium officinale is ripe gradually. The seedlings of Dendrobium officinale of current batch production is bred, being generally adopted Herba Dendrobii fruit pod is kind of a source, through artificial seeding's culture medium, after making embryo germination, it is about the 8-10 cultivation of individual month can obtain Herba Dendrobii finished product tissue cultured seedling through wooden dipper kind, single Seedling switching, strong sprout, four-stage of taking root. In single Seedling switching, strong sprout, take root in three phases switching process, artificial inoculation tweezers are needed frequently to be transferred to one by one in new culture medium by seedling, needing to expend substantial amounts of labour force and raw material in switching process, operation is loaded down with trivial details, inefficiency, causes that production cost remains high. Additionally in switching process, if dynamics control is improper, Seedling body physical damnification can be caused when gripping seedling, cause the situations such as Seedling bulk-growth is slow, yellowing leaf, overall chlorosis are dead, have a strong impact on the survival rate of seedling.
Summary of the invention
The technical problem to be solved is, for the deficiencies in the prior art, it is provided that a kind of simple, efficient candidum tissue culturing inoculation method.
This invention address that the technical scheme that above-mentioned technical problem adopts is: a kind of candidum tissue culturing inoculation method, comprise the following steps:
1) selection of Herba Dendrobii fruit pod: the Herba Dendrobii selecting outdoor pseudo-wild cultivating 3-6 raw is tied and grown the maturation formed 110-130 days and do not ftracture really pod;
2) embryo sowing: by Herba Dendrobii fruit pod on superclean bench after the alcohol disinfecting 12-15min that mass percent concentration is 65-75%, then the embryo in fruit pod is seeded in sterile beaker;
3) preparation of aseptic embryo mixed liquor: in step 2) sterile beaker in add sterilized water stirring, embryo is dispersed in sterilized water, dispersion is 60-100/mL, obtains aseptic embryo mixed liquor;
4) preparation of root media is sprouted: by culture medium based on 1/2MS culture medium, stereometer with this basal medium, following additive is added: naphthalene acetic acid 0.7-0.9mg/L in this basal medium, activated carbon 0.2-0.5g/L, 6-benzyl aminoadenine 0.05-0.2mg/L, coconut juice 50-80g/L, peeling Fructus Musae 30-50g/L, rice 20-30g/L, sucrose 25-35g/L and agar 5.8-6.3g/L, obtain sprouting root media, and the pH value of this sprouting root media is adjusted to 6.0 ~ 6.2, afterwards the root media of sprouting of preparation is dispensed into in the 650mL tissue culture bottle of air-vent, the liquid filling degree of depth of each tissue culture bottle is 3-4cm, then by each tissue culture bottle in high-pressure sterilizing pot in 121 DEG C of sterilizing 20-23min, after cooling standby,
5) inoculation of embryo and cultivation: being inoculated into by aseptic embryo mixed liquor sterile pipette in sprouting root media, the inoculum concentration of each tissue culture bottle is 800-1000 �� L, then be coated with aseptic spreading rod and put down, by the bottle cap screwing of tissue culture bottle, inoculation completes; Afterwards by each tissue culture bottle dark treatment 5-7 days at 22-24 DEG C, then every day intensity of illumination be 1200-1500Lux, light application time be 10-11h culture environment under cultivate 30-40 days, last every day intensity of illumination be 2500-3000Lux, light application time be 12-13h culture environment under cultivate some skies, until embryo germination obtain candidum tissue culturing seedling.
MS basal medium is currently used most common culture medium, its composition includes a great number of elements, trace element, iron salt, organic principle, inositol etc., 1/2MS basal medium and a great number of elements become the culture medium of original 1/2, it is possible to self-control or selection commercially available prod.
Preferably, in step 3), the defining method of the dispersion of embryo is: pipette the aseptic embryo mixed liquor of 1mL with liquid-transfering gun, after diluting 10 times with sterilized water, embryo is counted, it is determined that its dispersion, if dispersion is unsatisfactory for the requirement of 60-100/mL, in sterile beaker, then supplement embryo or sterilized water, until dispersion meets requirement.
Compared with prior art, it is an advantage of the current invention that: candidum tissue culturing inoculation method of the present invention, it is a kind of simple, efficient group training inoculation method, the method is inoculated in cultivation embryo through once inducing sprouting root media, it is made without switching repeatedly, and the sprouting root media of the inventive method preparation is based on natural bacteriostatic additive, add without harmful chemical component, it is thus possible to it is long by an inoculation debugging time effectively to solve Traditional Man, workload is big, pollution rate is high, the problem of the seedling poor stability produced, the inventive method can be greatly reduced the pollution rate in candidum tissue culturing process, reduce the group training cost of Herba Dendrobii, candidum tissue culturing inoculation method of the present invention makes candidum tissue culturing inoculation efficiency and transplanting survival rate be greatly improved, and production efficiency is 3-4 times of traditional method, and transplanting survival rate is up to more than 92-93%.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is described in further detail.
The candidum tissue culturing inoculation method of embodiment 1, comprises the following steps:
1) selection of Herba Dendrobii fruit pod: select the Herba Dendrobii of outdoor pseudo-wild cultivating life in 3 years to be tied and grow the maturation formed 110 days not ftracture really pod;
2) embryo sowing: by Herba Dendrobii fruit pod on superclean bench after the alcohol disinfecting 12min that mass percent concentration is 65%, then the embryo in fruit pod is seeded in sterile beaker;
3) preparation of aseptic embryo mixed liquor: in step 2) sterile beaker in add sterilized water stirring, embryo is dispersed in sterilized water, dispersion is 60-70/mL, obtains aseptic embryo mixed liquor;
4) preparation of root media is sprouted: by culture medium based on 1/2MS culture medium, stereometer with this basal medium, following additive is added: naphthalene acetic acid (NAA) 0.75mg/L in this basal medium, activated carbon 0.3g/L, 6-benzyl aminoadenine (6-BA) 0.08mg/L, coconut juice 50g/L, peeling Fructus Musae 35g/L, rice 22g/L, sucrose 25g/L and agar 5.8g/L, obtain sprouting root media, and the pH value of this sprouting root media is adjusted to 6.0, afterwards the root media of sprouting of preparation is dispensed into in the 650mL tissue culture bottle of air-vent, the liquid filling degree of depth of each tissue culture bottle is 3cm, then by each tissue culture bottle in high-pressure sterilizing pot in 121 DEG C of sterilizing 20min, after cooling standby,
5) inoculation of embryo and cultivation: being inoculated into by aseptic embryo mixed liquor sterile pipette in sprouting root media, the inoculum concentration of each tissue culture bottle is 800 �� L, then be coated with aseptic spreading rod and put down, by the bottle cap screwing of tissue culture bottle, inoculation completes; Afterwards by each tissue culture bottle dark treatment 5 days at 22 DEG C, then every day intensity of illumination be 1200-1500Lux, light application time be 10-11h culture environment under cultivate 30 days, last every day intensity of illumination be 2500-3000Lux, light application time be 12-13h culture environment under cultivate 130 days after, obtain candidum tissue culturing seedling, tissue cultured seedling is evenly distributed, Seedling body healthy and strong, transplanting survival rate is 92.3%, and aberration rate is zero.
The candidum tissue culturing inoculation method of embodiment 2, comprises the following steps:
1) selection of Herba Dendrobii fruit pod: select the Herba Dendrobii of outdoor pseudo-wild cultivating life in 5 years to be tied and grow the maturation formed 120 days not ftracture really pod;
2) embryo sowing: by Herba Dendrobii fruit pod on superclean bench after the alcohol disinfecting 14min that mass percent concentration is 70%, then the embryo in fruit pod is seeded in sterile beaker;
3) preparation of aseptic embryo mixed liquor: in step 2) sterile beaker in add sterilized water stirring, embryo is dispersed in sterilized water, dispersion is 75-85/mL, obtains aseptic embryo mixed liquor;
4) preparation of root media is sprouted: by culture medium based on 1/2MS culture medium, stereometer with this basal medium, following additive is added: naphthalene acetic acid (NAA) 0.8mg/L in this basal medium, activated carbon 0.35g/L, 6-benzyl aminoadenine (6-BA) 0.075mg/L, coconut juice 65g/L, peeling Fructus Musae 40g/L, rice 25g/L, sucrose 30g/L and agar 6.0g/L, obtain sprouting root media, and the pH value of this sprouting root media is adjusted to 6.1, afterwards the root media of sprouting of preparation is dispensed into in the 650mL tissue culture bottle of air-vent, the liquid filling degree of depth of each tissue culture bottle is 3.5cm, then by each tissue culture bottle in high-pressure sterilizing pot in 121 DEG C of sterilizing 22min, after cooling standby,
5) inoculation of embryo and cultivation: being inoculated into by aseptic embryo mixed liquor sterile pipette in sprouting root media, the inoculum concentration of each tissue culture bottle is 900 �� L, then be coated with aseptic spreading rod and put down, by the bottle cap screwing of tissue culture bottle, inoculation completes; Afterwards by each tissue culture bottle dark treatment 6 days at 23 DEG C, then every day intensity of illumination be 1200-1500Lux, light application time be 10-11h culture environment under cultivate 32 days, last every day intensity of illumination be 2500-3000Lux, light application time be 12-13h culture environment under cultivate 118 days after, obtain candidum tissue culturing seedling, tissue cultured seedling is evenly distributed, Seedling body healthy and strong, transplanting survival rate is 93.2%, and aberration rate is zero.
The candidum tissue culturing inoculation method of embodiment 3, comprises the following steps:
1) selection of Herba Dendrobii fruit pod: select the Herba Dendrobii of outdoor pseudo-wild cultivating life in 6 years to be tied and grow the maturation formed 130 days not ftracture really pod;
2) embryo sowing: by Herba Dendrobii fruit pod on superclean bench after the alcohol disinfecting 14min that mass percent concentration is 75%, then the embryo in fruit pod is seeded in sterile beaker;
3) preparation of aseptic embryo mixed liquor: in step 2) sterile beaker in add sterilized water stirring, embryo is dispersed in sterilized water, dispersion is 90-100/mL, obtains aseptic embryo mixed liquor;
4) preparation of root media is sprouted: by culture medium based on 1/2MS culture medium, stereometer with this basal medium, following additive is added: naphthalene acetic acid (NAA) 0.9mg/L in this basal medium, activated carbon 0.4g/L, 6-benzyl aminoadenine (6-BA) 0.1mg/L, coconut juice 75g/L, peeling Fructus Musae 45g/L, rice 30g/L, sucrose 33g/L and agar 6.2g/L, obtain sprouting root media, and the pH value of this sprouting root media is adjusted to 6.2, afterwards the root media of sprouting of preparation is dispensed into in the 650mL tissue culture bottle of air-vent, the liquid filling degree of depth of each tissue culture bottle is 4cm, then by each tissue culture bottle in high-pressure sterilizing pot in 121 DEG C of sterilizing 23min, after cooling standby,
5) inoculation of embryo and cultivation: being inoculated into by aseptic embryo mixed liquor sterile pipette in sprouting root media, the inoculum concentration of each tissue culture bottle is 1000 �� L, then be coated with aseptic spreading rod and put down, by the bottle cap screwing of tissue culture bottle, inoculation completes; Afterwards by each tissue culture bottle dark treatment 5 days at 24 DEG C, then every day intensity of illumination be 1200-1500Lux, light application time be 10-11h culture environment under cultivate 35 days, last every day intensity of illumination be 2500-3000Lux, light application time be 12-13h culture environment under cultivate 115 days after, obtain candidum tissue culturing seedling, tissue cultured seedling is evenly distributed, Seedling body healthy and strong, transplanting survival rate is 93.5%, and aberration rate is zero.
In the step 3) of above example 1-3, following methods can be adopted to determine the dispersion of embryo: pipette the aseptic embryo mixed liquor of 1mL with liquid-transfering gun, after diluting 10 times with sterilized water, embryo is counted, determine its dispersion, if dispersion is unsatisfactory for the requirement of 60-100/mL, then in sterile beaker, supplement embryo or sterilized water, until dispersion meets requirement.
Claims (2)
1. a candidum tissue culturing inoculation method, it is characterised in that comprise the following steps:
1) selection of Herba Dendrobii fruit pod: the Herba Dendrobii selecting outdoor pseudo-wild cultivating 3-6 raw is tied and grown the maturation formed 110-130 days and do not ftracture really pod;
2) embryo sowing: by Herba Dendrobii fruit pod on superclean bench after the alcohol disinfecting 12-15min that mass percent concentration is 65-75%, then the embryo in fruit pod is seeded in sterile beaker;
3) preparation of aseptic embryo mixed liquor: in step 2) sterile beaker in add sterilized water stirring, embryo is dispersed in sterilized water, dispersion is 60-100/mL, obtains aseptic embryo mixed liquor;
4) preparation of root media is sprouted: by culture medium based on 1/2MS culture medium, stereometer with this basal medium, following additive is added: naphthalene acetic acid 0.7-0.9mg/L in this basal medium, activated carbon 0.2-0.5g/L, 6-benzyl aminoadenine 0.05-0.2mg/L, coconut juice 50-80g/L, peeling Fructus Musae 30-50g/L, rice 20-30g/L, sucrose 25-35g/L and agar 5.8-6.3g/L, obtain sprouting root media, and the pH value of this sprouting root media is adjusted to 6.0 ~ 6.2, afterwards the root media of sprouting of preparation is dispensed into in the 650mL tissue culture bottle of air-vent, the liquid filling degree of depth of each tissue culture bottle is 3-4cm, then by each tissue culture bottle in high-pressure sterilizing pot in 121 DEG C of sterilizing 20-23min, after cooling standby,
5) inoculation of embryo and cultivation: being inoculated into by aseptic embryo mixed liquor sterile pipette in sprouting root media, the inoculum concentration of each tissue culture bottle is 800-1000 �� L, then be coated with aseptic spreading rod and put down, by the bottle cap screwing of tissue culture bottle, inoculation completes; Afterwards by each tissue culture bottle dark treatment 5-7 days at 22-24 DEG C, then every day intensity of illumination be 1200-1500Lux, light application time be 10-11h culture environment under cultivate 30-40 days, last every day intensity of illumination be 2500-3000Lux, light application time be 12-13h culture environment under cultivate some skies, until embryo germination obtain candidum tissue culturing seedling.
2. a kind of candidum tissue culturing inoculation method according to claim 1, it is characterized in that in step 3), the defining method of the dispersion of embryo is: pipette the aseptic embryo mixed liquor of 1mL with liquid-transfering gun, after diluting 10 times with sterilized water, embryo is counted, it is determined that its dispersion, if dispersion is unsatisfactory for the requirement of 60-100/mL, in sterile beaker, then supplement embryo or sterilized water, until dispersion meets requirement.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610046856.1A CN105613296A (en) | 2016-01-25 | 2016-01-25 | Tissue culture inoculating method for dendrobium officinale |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610046856.1A CN105613296A (en) | 2016-01-25 | 2016-01-25 | Tissue culture inoculating method for dendrobium officinale |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105613296A true CN105613296A (en) | 2016-06-01 |
Family
ID=56029009
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610046856.1A Pending CN105613296A (en) | 2016-01-25 | 2016-01-25 | Tissue culture inoculating method for dendrobium officinale |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105613296A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109793010A (en) * | 2019-03-14 | 2019-05-24 | 华侨大学 | A kind of liquid composite bacteria agent and its application method promoting Growth of Dendrobium candidum |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1961632A (en) * | 2006-12-08 | 2007-05-16 | 蔡仲玉 | An officinal dendrobium stem plantation method |
| CN102612966A (en) * | 2012-04-05 | 2012-08-01 | 中国科学院西双版纳热带植物园 | Method for obtaining dendrobium aphyllum seeds to germinate effective symbiotic fungi |
| CN102612965A (en) * | 2012-04-05 | 2012-08-01 | 中国科学院西双版纳热带植物园 | Method for obtaining germination effective symbiotic fungus for Cymbidium bicolor seeds |
| CN102845308A (en) * | 2012-09-27 | 2013-01-02 | 浙江省中药研究所有限公司 | Method for dendrobium officinale tissue culture one-step seedling |
| CN103392602A (en) * | 2013-08-09 | 2013-11-20 | 中国科学院华南植物园 | Sterile seeding and test-tube plantlet propagation method of sword-leaved cymbidiums |
| CN105028213A (en) * | 2015-09-01 | 2015-11-11 | 宣威市农硕农特产品开发有限公司 | Tissue-culturing rapid propagation method for dendrobium officinale |
| CN105104209A (en) * | 2015-09-24 | 2015-12-02 | 戴亚峰 | Method for cultivating tissues to obtain seedlings of dendrobium huoshanense at one step and formula of nutrient liquid for method |
-
2016
- 2016-01-25 CN CN201610046856.1A patent/CN105613296A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1961632A (en) * | 2006-12-08 | 2007-05-16 | 蔡仲玉 | An officinal dendrobium stem plantation method |
| CN102612966A (en) * | 2012-04-05 | 2012-08-01 | 中国科学院西双版纳热带植物园 | Method for obtaining dendrobium aphyllum seeds to germinate effective symbiotic fungi |
| CN102612965A (en) * | 2012-04-05 | 2012-08-01 | 中国科学院西双版纳热带植物园 | Method for obtaining germination effective symbiotic fungus for Cymbidium bicolor seeds |
| CN102845308A (en) * | 2012-09-27 | 2013-01-02 | 浙江省中药研究所有限公司 | Method for dendrobium officinale tissue culture one-step seedling |
| CN103392602A (en) * | 2013-08-09 | 2013-11-20 | 中国科学院华南植物园 | Sterile seeding and test-tube plantlet propagation method of sword-leaved cymbidiums |
| CN105028213A (en) * | 2015-09-01 | 2015-11-11 | 宣威市农硕农特产品开发有限公司 | Tissue-culturing rapid propagation method for dendrobium officinale |
| CN105104209A (en) * | 2015-09-24 | 2015-12-02 | 戴亚峰 | Method for cultivating tissues to obtain seedlings of dendrobium huoshanense at one step and formula of nutrient liquid for method |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109793010A (en) * | 2019-03-14 | 2019-05-24 | 华侨大学 | A kind of liquid composite bacteria agent and its application method promoting Growth of Dendrobium candidum |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103843662B (en) | A kind of promote Herba Dendrobii tissue cultured seedling strong sprout and the method taken root | |
| CN102405842B (en) | Open type method for cultivating toxin-free seedlings of sugarcanes | |
| CN104885941B (en) | Chemical disinfection tissue culture method for miscanthus giganteus | |
| CN104380994A (en) | Bletilla striata large-scale high-yield cultivation method | |
| CN103999776A (en) | Orchid rapid-propagation culture medium formula | |
| CN101836585B (en) | Tissue-culture seedling raising method of rhodiola crenulata | |
| CN102823497A (en) | Clonal tissue culture breeding method of Liquidambar formosana Hance | |
| CN104472366A (en) | Tissue culture rapid-propagation method for improving salt tolerance of seedlings of southern ecotype jujubes | |
| CN101946709A (en) | Seedling raising method of plateau rhodiola crenulata | |
| CN102138532A (en) | Technique for breeding seedlings of Taihe tribute Chinese toon by tissue culture | |
| CN104488723A (en) | Tissue-culture and rapid-propagation method of epimedium koreanum nakai | |
| CN105660400A (en) | Strengthening and weight increasing method for anoectochilus roxburghii tissue cultured seedlings | |
| CN105766636B (en) | A kind of peony tissue culture regeneration method | |
| CN104322371A (en) | Tissue culture medium and tissue culture method of dendrobium officinale | |
| CN103238519A (en) | Rapid seedling raising method of switchgrass tissue culture | |
| CN104094848A (en) | Induction of tung tree hypocotyls callus and method for efficiently regenerating plants | |
| CN106386504A (en) | Tissue culture method of Aralia Cordata Thunb seedlings | |
| CN103636506A (en) | Shepherdia argentea caulicle regenerated plant induction culture medium and method for performing plant culture by utilizing culture medium | |
| CN103477750A (en) | Method for promoting germination of rosemary seed | |
| CN103931499B (en) | A kind of quick breeding method for tissue culture of callistemon rigidus | |
| CN105766639A (en) | Tissue culture and quick seedling culture method of sweet sorghum | |
| CN105613296A (en) | Tissue culture inoculating method for dendrobium officinale | |
| CN105145042A (en) | Soil cultivation method for potato virus-free basic seeds | |
| CN104813933A (en) | Method for culturing yam seed tuber by using yam tissue culture seedling | |
| CN104686342A (en) | Asexual and rapid propagation technology for styrax tonkinensis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160601 |
|
| RJ01 | Rejection of invention patent application after publication |