CN104542255A - Method for solving flowering asynchronism during crossbreeding of cymbidium - Google Patents
Method for solving flowering asynchronism during crossbreeding of cymbidium Download PDFInfo
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- CN104542255A CN104542255A CN201410857115.2A CN201410857115A CN104542255A CN 104542255 A CN104542255 A CN 104542255A CN 201410857115 A CN201410857115 A CN 201410857115A CN 104542255 A CN104542255 A CN 104542255A
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Abstract
The invention discloses a method for solving flowering asynchronism during crossbreeding of cymbidium. The method includes the following steps: picking out an anther cap of cymbidium through a sterilized picking tool when the petal of a cymbidium simple flower unfolds; picking the cymbidium pollinium to a clean and sterile container through the sterilized picking tool and sealing the pollinium in the sterile container; storing the container in a dark place at the temperature of 4-6 DEG C; unfreezing the stored pollinium at the room temperature for 20-30 min when the female parent orchid blooms; adding sterile water and performing rewetting for 20-30 min to obtain the vital pollen. According to the invention, the cymbidium pollinium stored as per the method provided by the invention can be stored for 3-12 months without going mouldy; after unfreezing and rewetting, and measured by the 2,3,5-triphenyltetrazolium chloride staining method, the vitality of the pollen stored according to the method is high, the success rate of hand pollination is high, and the hybridization success rate is high.
Description
Technical field:
The invention belongs to field of plant reproduction, be specifically related to a kind of method solving orchid crossbreeding flowering asynchronism.
Background technology:
Orchid is the rare flower in the fashionable world, is to cultivate one of the widest, sales volume is maximum flowers kind in the world, and development in recent years is particularly rapid, every year with more than 10% speed increment, current whole world orchid year the amount of consumption more than 4,000,000,000 dollars.Holland, Thailand, Singapore, the U.S., Korea S, Japan etc. are the more flourishing countries of orchid industry, and Thailand is the maximum exported country of cattleya.The orchid of current production mostly is the cattleyas such as Moth orchid, dendrobium and Bowring cattleya, and the state orchid with Chinese tradition characteristic is then small.China is one of maximum producing country of orchid at present, our province is again the main orchid production base of China and product distributing center, but the main cymbidium variety of the current large-scale production of our province and even China, as Moth orchid, the provenance such as hybrid cymbidium and the stem of noble dendrobium is substantially still from Taiwan, the ground such as Southeast Asia are introduced, in these kinds introduced, some kind is also not exclusively applicable to our province cultivation, in addition due to the renewal dependence on import of provenance, Cultivar replacing is caused to be regenerated slow, provenance serious degradation, production cost is high, the height of flowers price also has very large relation with its novelty simultaneously, and mostly the flower variety introduced is to plant for many years being introduced into state (area), novelty greatly reduces, this present situation has made the development of the orchid industry of our province and the raising of benefit receive serious restriction.
Crossbreeding is the effective means of cultivating orchid new varieties, but there is flowering asynchronism bottleneck problem when orchid hybridizes, and cannot carry out some distant hybridization due to flowering asynchronism simultaneously.Flowering asynchronism is the bottleneck problem in orchid crossbreeding, effective preservation of pollen is one of method solving flowering asynchronism, low temperature, the Excised Embryos technology of orchid pollen have been reported, but exist Pollen Activity decline fast, preservation effect is poor, preserve the problem and cause pollen to preserve unsuccessfully of often going mouldy, or the shortcoming such as complex operation, preservation condition requirement difficulty be satisfied.
Summary of the invention:
The object of this invention is to provide a kind of effective preservation that can meet orchid pollinium under simple laboratory condition, the holding time of pollinium reaches 1 year and still has fine vigor, and can not the problem of going mouldy be produced, the method for the solution orchid crossbreeding flowering asynchronism of the requirement of the orchid crossbreeding in different flowering period can be met at any time.
The method of solution orchid crossbreeding flowering asynchronism of the present invention, it is characterized in that, when singly petal just launches orchid, the picking instrument of application sterilizing picks out colored anther cap, with the picking instrument of this sterilizing, pollinium is chosen good seal to the container of cleaning sterile again, be placed in 4 ~ 6 DEG C keep in Dark Place (time of orchid pollen preservation can reach 3 ~ 12 months), when maternal orchid blooms, thaw the pollinium of above-mentioned preservation 20 ~ 30min at ambient temperature, and then add sterile water and carry out rewetting process 20 ~ 30min, obtain the great-hearted pollen of tool.
The great-hearted pollen of tool may be used for artificial pollination on maternal column cap, then through cultivating allocarpy, the link such as aseptic seeding and test-tube seedling transplanting, obtains orchid hybridization seedling, thus the effective problem solving orchid crossbreeding flowering asynchronism.
Preferably, the method of described solution orchid crossbreeding flowering asynchronism is when singly petal just launches orchid, in the morning 8 ~ 9 time, apply sterilizing toothpick pick out colored anther cap gently, pollinium to be sticked on toothpick point with sterilizing toothpick again and rapidly pollinium put in the 1ml EP pipe of cleaning sterile, often the pollinium of 1 flower put into by pipe, manage to concentrate with an aseptic vial again load also good seal by placed pollinium EP, be placed in 4 ~ 6 DEG C keep in Dark Place (time of orchid pollen preservation can reach 3 ~ 12 months), when maternal orchid blooms, thaw the pollinium of above-mentioned Cord blood 20 ~ 30min at ambient temperature, and then add several sterile waters and carry out rewetting process 20 ~ 30 minutes in the EP pipe placing pollinium, obtain the great-hearted pollen of tool.This preferred version can be preserved by bonding flower single tube, thus ensures the selectivity of artificial pollination.
Utilize the orchid pollinium that method of the present invention is preserved, the time that its orchid pollen is preserved can reach 3 ~ 12 months, and can not be mouldy, after separating the process of cold-peace rewetting, find through TTC Determination Staining pollen viability, the Pollen Activity that this store method is preserved is strong, and artificial pollination success rate is high, is hybridized to power high.And the instrument that method of the present invention adopts is easier, storage condition is simple, only need leave the preservation by low temperature layer of general refrigerator in just.Therefore by the enforcement of patent of the present invention, the hybridization pollination of the kind and plant that solve different female bloom dates hinders, be expected to make a breakthrough at orchid rearing new variety, thus can be China's orchid rearing new variety and further developing of industry lays the foundation, strengthen the competitiveness of China in world's industry of flowers and plants, and obtain good economic benefit and social benefit.
Embodiment:
Following examples further illustrate of the present invention, instead of limitation of the present invention.
What use in embodiment spends precious No. 1 (HYPONeX 1) to be Taiwan produced in USA and gardening enterprise stock action packing product.
Embodiment 1: the crossbreeding of Doritis pulcherrima (Doritis pulcherrima) and Yunnan Herba Renantherae coccineae (Renanthera imschootiana) and sapling multiplication
(1) artificial pollination: Yunnan Herba Renantherae coccineae was bloomed at the beginning of 3 months, pollen is gathered when its single flower petal just launches, in the morning 8 ~ 9 time, apply sterilized toothpick pick out colored anther cap gently, pollinium to be sticked on toothpick point with the toothpick of sterilizing again and rapidly pollinium put in the 1ml EP pipe of cleaning sterile, often the pollinium of 1 flower put into by pipe, manage to concentrate with an aseptic vial again load also good seal by placed pollinium EP, keep in Dark Place under finally the vial filling pollen being positioned over 4 DEG C of conditions (refrigerator preservation by low temperature layer).The time that Herba Renantherae coccineae pollen is preserved when maternal Doritis pulcherrima is in time blooming at the beginning of 6 months then, thaws and rewetting process to the pollen of the Herba Renantherae coccineae of above-mentioned Cord blood, to recover and to improve the vitality of pollen after reaching 3 months.First carry out rewetting process 20 minutes by adding several sterile waters again in the EP pipe placing pollinium after the thawing at ambient temperature 20 minutes of the Herba Renantherae coccineae pollen of Cord blood, finally adopt TTC Determination Staining pollen viability, rate of dyeing reaches 71.5%, determine that pollen has vigor, invested on maternal column cap.Bagging after the lip that after pollination, excision is maternal, removes bag after about 1 week, hang up label, label has the information such as father, maternal title, hybridization date and pollination people.Note observing its developmental state, the success rate of statistics pollination when 3 months, ripening rate all reaches more than 90%, can gather and carry out aseptic seeding when waiting fruit to become orange-yellow.
(2) aseptic seeding: the pollination fruit of latter 5 months is first put on the skin with alcohol cotton and wipes clean fruit surface dirt, then within alcohol-pickled 2 minutes, be placed in the mercuric chloride solution of 0.1% with 70% and sterilize 30 minutes, fruit is cut after rinsed with sterile water 5 times, Powdered embryo is inoculated on seed germination medium, sterilization success rate more than 95%, when 20 days, visible seed expands and turns green, and when 40 days, seed germination forms protocorm, within 90 days, forms seedling further.Seed germination rate 90%, planting percent 90%.Seed germination medium often rises containing spending precious No. 1 2.0g, peptone 2.0g, coconut milk 100mL, sucrose 20g, agar 6g, active carbon 1.0g, and all the other are vitamin in MS medium and inositol composition, Yi Jishui; PH 5.4 ~ 5.6.
(3) strong seedling culture: the seedling inoculated and cultured obtained by aseptic seeding, on strong seedling culture base, can form healthy and strong plant when 120 days.Described strong seedling culture base often rises containing spending precious No. 1 3.0g, peptone 2.0g, methyl α-naphthyl acetate 1mg, banana homogenate 80g, coconut milk 50mL, sucrose 15g, agar 6g, active carbon 2.0g, and all the other are vitamin in MS medium and inositol composition, Yi Jishui; PH 5.6 ~ 5.8.
(4) test-tube seedling transplanting: when plant is about 5-8 centimetre high, blake bottle is transferred to natural daylight lower refining seedling 15 days, then it is taken out from vial, clean the medium of root, move into blue stone: wood chip: bark volume ratio is in the mixed-matrix of 1:1:1, keep suitably ventilating and enough humidity, the survival rate of transplanting all can reach more than 95%.
Condition of culture used is in an experiment cultivation temperature 24 DEG C ~ 28 DEG C, illuminance 1500 ~ 2000lx, illumination 12 hours/day.
Embodiment 2: little Hua is the crossbreeding of blue (Vanda coerulescens) and Yunnan Herba Renantherae coccineae (Renanthera imschootiana) and sapling multiplication all ages
(1) artificial pollination: Yunnan Herba Renantherae coccineae was bloomed at the beginning of 3 months, pollen is gathered when its single flower petal just launches, in the morning 8 ~ 9 time, apply sterilized toothpick pick out colored anther cap gently, pollinium to be sticked on toothpick point with sterilizing toothpick again and rapidly pollinium put in the 1ml EP pipe of cleaning sterile, often the pollinium of 1 flower put into by pipe, manage to concentrate with an aseptic vial again load also good seal by placed pollinium EP, keep in Dark Place under finally the vial filling pollen being positioned over 4 DEG C of conditions.The time that Herba Renantherae coccineae pollen is preserved when maternal little Hua is all ages blue in time blooming at the beginning of 9 months then, thaws and rewetting process to the pollen of the Herba Renantherae coccineae of Cord blood, to recover and to improve the vitality of pollen after reaching 6 months.In the EP pipe of placement pollinium, add several sterile waters again after first being thawed at ambient temperature 25 minutes by the Herba Renantherae coccineae pollen of Cord blood and carry out rewetting process 25 minutes, finally adopt TTC Determination Staining pollen viability, rate of dyeing reaches 65%, determine that pollen has vigor, invested on maternal column cap.Bagging after the lip that after pollination, excision is maternal, removes bag after about 1 week, hang up label, label has the information such as father, maternal title, hybridization date and pollination people.Note observing its developmental state, the success rate of statistics pollination when 3 months, ripening rate all reaches more than 80%, can gather and carry out aseptic seeding when waiting fruit to become orange-yellow.
(2) aseptic seeding: the pollination fruit of latter 5 months is first put on the skin with alcohol cotton and wipes clean fruit surface dirt, then within alcohol-pickled 3 minutes, be placed in the mercuric chloride solution of 0.2% with 75% and sterilize 20 minutes, fruit is cut after rinsed with sterile water 5 times, Powdered embryo is inoculated on seed germination medium, sterilization success rate reaches 100%, visible seed germination when 30 days, when 60 days, seed germination forms protocorm, within 120 days, forms seedling further.Seed germination rate 80%, planting percent 60%.Seed germination medium often rises containing spending precious No. 1 3.0g, peptone 1.5g, coconut milk 150mL, sucrose 20g, agar 6.5g, active carbon 0.5g, and all the other are vitamin in MS medium and inositol composition, Yi Jishui; PH 5.4 ~ 5.6.
(3) strong seedling culture: the seedling inoculated and cultured obtained by aseptic seeding, on strong seedling culture base, can form healthy and strong plant when 150 days.Described strong seedling culture base often rises containing spending precious No. 1 3.0g, peptone 1.5g, methyl α-naphthyl acetate 0.5mg, banana homogenate 100g, coconut milk 100mL, sucrose 20g, agar 7g, active carbon 1.0g, and all the other are vitamin in MS medium and inositol composition, Yi Jishui; PH 5.6 ~ 5.8.
(4) test-tube seedling transplanting: when plant about 5 ~ 8 centimetres is high, blake bottle is transferred to natural daylight lower refining seedling 20 days, then it is taken out from vial, clean the medium of root, move into blue stone: wood chip: bark volume ratio is in the mixed-matrix of 1:1:1, keep suitably ventilating and enough humidity, the survival rate of transplanting all can reach more than 90%.
Condition of culture used is in an experiment cultivation temperature 24 ~ 28 DEG C, illuminance 1500 ~ 2000lx, illumination 12 hours/day.
Embodiment 3: the crossbreeding of color heart sword-leaved cymbidium (Cymbidium ensifolium) and Yunnan Herba Renantherae coccineae (Renanthera imschootiana) and sapling multiplication
(1) artificial pollination: Yunnan Herba Renantherae coccineae was bloomed at the beginning of 3 months, pollen is gathered when its single flower petal just launches, in the morning 8 ~ 9 time, apply sterilized toothpick pick out colored anther cap gently, pollinium to be sticked on toothpick point with toothpick again and rapidly pollinium put in the 1ml EP pipe of cleaning sterile, often the pollinium of 1 flower put into by pipe, manage to concentrate with an aseptic vial again load also good seal by placed pollinium EP, keep in Dark Place under finally the vial filling pollen being positioned over 6 DEG C of conditions.The time that Herba Renantherae coccineae pollen is preserved when the color heart sword-leaved cymbidium of female parent is in time blooming at the beginning of 12 months then, thaws and rewetting process to the pollen of the Herba Renantherae coccineae of Cord blood, to recover and to improve the vitality of pollen after reaching 9 months.In the EP pipe of placement pollinium, add several sterile waters again after first being thawed at ambient temperature 30 minutes by the Herba Renantherae coccineae pollen of Cord blood and carry out rewetting process 30 minutes, finally adopt TTC Determination Staining pollen viability, rate of dyeing reaches 56%, determine that pollen has vigor, invested on maternal column cap.Bagging after the lip that after pollination, excision is maternal, removes bag after about 1 week, hang up label, label has the information such as father, maternal title, hybridization date and pollination people.Note observing its developmental state, the success rate of statistics pollination when 3 months, ripening rate reaches 30%, can gather and carry out aseptic seeding when waiting fruit to become orange-yellow.
(2) aseptic seeding: the pollination fruit of latter 12 months is first put on the skin with alcohol cotton and wipes clean fruit surface dirt, then within alcohol-pickled 3 minutes, be placed in the mercuric chloride solution of 0.2% with 70% and sterilize 30 minutes, fruit is cut after rinsed with sterile water 4 times, Powdered embryo is inoculated on seed germination medium, sterilization success rate reaches 100%, visible seed germination when 90 days, when 120 days, seed germination forms root-like stock, by root-like stock inoculated and cultured in differential medium, within 60 days, seedling can be differentiated to form.Seed germination rate 70%, planting percent 65%.Seed germination medium often rises containing spending precious No. 1 1.5g, peptone 1.5g, coconut milk 50mL, sucrose 15g, agar 6g, active carbon 1.5g, and all the other are vitamin in MS medium and inositol composition, Yi Jishui; PH 5.8-6.0.Differential medium is often liter of interpolation 6-Bian Ji adenine 5.0mg on germination medium basis.
(3) strong seedling culture: seedling inoculated and cultured root-like stock differentiation obtained, on strong seedling culture base, can form healthy and strong plant when 150 days.Described strong seedling culture base often rises containing spending precious No. 1 1.5g, peptone 1.5g, methyl α-naphthyl acetate 0.5mg, banana homogenate 50g, coconut milk 50mL, sucrose 15g, agar 6g, active carbon 0.5g, and all the other are vitamin in MS medium and inositol composition, Yi Jishui; PH 5.6-5.8.
(4) test-tube seedling transplanting: when plant about 5 ~ 8 centimetres is high, blake bottle is transferred to natural daylight lower refining seedling 15 days, then it is taken out from vial, clean the medium of root, move into blue stone: wood chip: bark volume ratio is in the mixed-matrix of 1:1:1, keep suitably ventilating and enough humidity, the survival rate of transplanting all can reach more than 95%.
Condition of culture used is in an experiment cultivation temperature 24-28 DEG C, illuminance 1500 ~ 2000lx, illumination 12 hours/day.
Claims (2)
1. one kind solves the method for orchid crossbreeding flowering asynchronism, it is characterized in that, when singly petal just launches orchid, the picking instrument of application sterilizing picks out the anther cap of orchid, with the picking instrument of this sterilizing, orchid pollinium is chosen good seal to the container of cleaning sterile again, be placed in 4 ~ 6 DEG C to keep in Dark Place, when maternal orchid blooms, thaw the pollinium of above-mentioned preservation 20 ~ 30min at ambient temperature, and then add sterile water and carry out rewetting process 20 ~ 30min, obtain the great-hearted pollen of tool.
2. method according to claim 1, it is characterized in that, the method of described solution orchid crossbreeding flowering asynchronism is when singly petal just launches orchid, the anther cap that sterilizing toothpick picks out orchid is gently applied in the morning 8 ~ 9 time, orchid pollinium to be sticked on toothpick point with sterilizing toothpick again and rapidly pollinium put in the 1ml EP pipe of cleaning sterile, often the pollinium of 1 flower put into by pipe, manage to concentrate with an aseptic vial again load also good seal by placed pollinium EP, be placed in 4 ~ 6 DEG C to keep in Dark Place, when maternal orchid blooms, thaw the pollinium of above-mentioned Cord blood 20 ~ 30min at ambient temperature, and then add several sterile waters and carry out rewetting process 20 ~ 30 minutes in the EP pipe placing pollinium, obtain the great-hearted pollen of tool.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107466593A (en) * | 2017-08-22 | 2017-12-15 | 中国科学院华南植物园 | A kind of pocket orchid powder low temperature drying store method |
| CN108651444A (en) * | 2018-05-22 | 2018-10-16 | 山东省农业科学院蔬菜花卉研究所 | A kind of iris pollen preserves and breeding method |
| CN109089817A (en) * | 2018-08-30 | 2018-12-28 | 中国热带农业科学院热带作物品种资源研究所 | A kind of Herba Renantherae coccineae florescence control method |
| CN109122301A (en) * | 2018-10-19 | 2019-01-04 | 中国科学院华南植物园 | A kind of method of Doritis pulcherrima and Herba Renantherae coccineae generic cross cultivation orchid new varieties |
| CN109122302A (en) * | 2018-10-19 | 2019-01-04 | 中国科学院华南植物园 | A kind of method of sword-leaved cymbidium and Herba Renantherae coccineae generic cross cultivation orchid new varieties |
-
2014
- 2014-12-31 CN CN201410857115.2A patent/CN104542255A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| 程式君: "兰花花粉生命力保存的方法", 《园艺学报》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107466593A (en) * | 2017-08-22 | 2017-12-15 | 中国科学院华南植物园 | A kind of pocket orchid powder low temperature drying store method |
| CN108651444A (en) * | 2018-05-22 | 2018-10-16 | 山东省农业科学院蔬菜花卉研究所 | A kind of iris pollen preserves and breeding method |
| CN108651444B (en) * | 2018-05-22 | 2020-11-27 | 山东省农业科学院蔬菜花卉研究所 | Preservation and breeding method for phalaenopsis pollen |
| CN109089817A (en) * | 2018-08-30 | 2018-12-28 | 中国热带农业科学院热带作物品种资源研究所 | A kind of Herba Renantherae coccineae florescence control method |
| CN109122301A (en) * | 2018-10-19 | 2019-01-04 | 中国科学院华南植物园 | A kind of method of Doritis pulcherrima and Herba Renantherae coccineae generic cross cultivation orchid new varieties |
| CN109122302A (en) * | 2018-10-19 | 2019-01-04 | 中国科学院华南植物园 | A kind of method of sword-leaved cymbidium and Herba Renantherae coccineae generic cross cultivation orchid new varieties |
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Application publication date: 20150429 |