CN114128585A - Method for improving transplanting survival rate of rare or endangered orchid tissue culture seedlings - Google Patents
Method for improving transplanting survival rate of rare or endangered orchid tissue culture seedlings Download PDFInfo
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- CN114128585A CN114128585A CN202111464725.2A CN202111464725A CN114128585A CN 114128585 A CN114128585 A CN 114128585A CN 202111464725 A CN202111464725 A CN 202111464725A CN 114128585 A CN114128585 A CN 114128585A
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- soil
- orchid
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- 241000233855 Orchidaceae Species 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 18
- 230000004083 survival effect Effects 0.000 title claims abstract description 17
- 239000002689 soil Substances 0.000 claims abstract description 50
- 241000196324 Embryophyta Species 0.000 claims abstract description 46
- 239000000725 suspension Substances 0.000 claims abstract description 26
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 19
- 239000008103 glucose Substances 0.000 claims abstract description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 17
- 230000002068 genetic effect Effects 0.000 claims abstract description 16
- 238000012258 culturing Methods 0.000 claims abstract description 10
- 239000002002 slurry Substances 0.000 claims abstract description 10
- 239000000463 material Substances 0.000 claims abstract description 8
- 238000007796 conventional method Methods 0.000 claims abstract description 5
- 241000894007 species Species 0.000 claims abstract description 5
- 230000000474 nursing effect Effects 0.000 claims abstract description 3
- 230000001954 sterilising effect Effects 0.000 claims description 7
- 238000004659 sterilization and disinfection Methods 0.000 claims description 7
- 238000012268 genome sequencing Methods 0.000 claims description 4
- 238000007598 dipping method Methods 0.000 abstract description 4
- 238000006748 scratching Methods 0.000 abstract description 4
- 230000002393 scratching effect Effects 0.000 abstract description 4
- 238000002791 soaking Methods 0.000 abstract description 4
- 239000012153 distilled water Substances 0.000 description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- 239000000741 silica gel Substances 0.000 description 3
- 229910002027 silica gel Inorganic materials 0.000 description 3
- 230000009261 transgenic effect Effects 0.000 description 3
- 241000173289 Cymbidium goeringii Species 0.000 description 2
- 235000017141 Cymbidium goeringii Nutrition 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 235000017166 Bambusa arundinacea Nutrition 0.000 description 1
- 235000017491 Bambusa tulda Nutrition 0.000 description 1
- 235000007516 Chrysanthemum Nutrition 0.000 description 1
- 244000189548 Chrysanthemum x morifolium Species 0.000 description 1
- 241000732800 Cymbidium Species 0.000 description 1
- 241000721047 Danaus plexippus Species 0.000 description 1
- 244000082204 Phyllostachys viridis Species 0.000 description 1
- 235000015334 Phyllostachys viridis Nutrition 0.000 description 1
- 239000011425 bamboo Substances 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/60—Flowers; Ornamental plants
- A01G22/63—Orchids
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Cultivation Of Plants (AREA)
Abstract
The invention discloses a method for improving the transplanting survival rate of rare or endangered orchid tissue culture seedlings, which comprises the following steps: (1) performing germplasm resource survey on the orchid rare species; (2) analyzing the genetic diversity of the plants, and determining the population with the highest genetic diversity; (3) collecting rhizosphere soil and overground part orchid, and storing the rhizosphere soil and plant materials; (4) placing rhizosphere soil in a bottle, mixing the soil and water, adding a glucose solution, and culturing to obtain a soil suspension; (5) mixing the plants, grinding and crushing, placing in a bottle, adding water, adding a glucose solution, and culturing to obtain a plant suspension; (6) mixing the soil suspension with common soil to prepare slurry; (7) scratching the stem part of the hardened orchid tissue culture seedling, soaking the stem part in plant suspension, and dipping the root in slurry; (8) and (4) planting or returning and nursing according to a conventional method. The method greatly improves the self resistance of the orchid, and further improves the transplanting survival rate of the wild orchid.
Description
Technical Field
The invention relates to the technical field of flower cultivation, in particular to a method for improving the transplanting survival rate of rare or endangered orchid tissue culture seedlings.
Background
Orchid is an orchid plant widely distributed in various parts of China. The orchid cultivated in China has a history of more than two thousand years, and the orchid has been regarded as a high-cleanness and elegant symbol all the time, is parallel to the plum, bamboo and chrysanthemum, is called as 'four monarch drugs' collectively, and is one of ten famous flowers in China. Orchids are generally epiphytic or terrestrial herbaceous plants, and the common planting methods are to fix the orchids on trunks (or barks) and to plant the orchids in a matrix, wherein the cultivation and the matrix are common. However, because of poor adaptability of rare orchid species, the survival rate of many endangered orchid species after transplantation is low. Therefore, how to invent a planting method which can plant orchid on log and has high beauty and high survival rate of orchid becomes a difficult problem which needs to be solved urgently in the field of orchid planting.
Disclosure of Invention
The invention aims to solve the technical problem of providing a method for improving the transplanting survival rate of rare or endangered orchid tissue culture seedlings.
In order to achieve the purpose, the technical scheme provided by the invention is as follows:
a method for improving the transplanting survival rate of rare or endangered orchid tissue culture seedlings comprises the following operation steps:
(1) firstly, performing germplasm resource survey on rare species of orchid, collecting young leaves of 5-10 plants (two plants are more than 5 meters apart) from each colony, and storing in a sealed bag filled with silica gel;
(2) analyzing the genetic diversity of the transgenic plant by adopting molecular means such as ISSR, RAPD, simplified genome sequencing and the like, and determining the population with the highest genetic diversity;
(3) collecting rhizosphere soil and overground part orchid (not requiring the whole plant and having a length of 5-10 cm) of the population with the highest genetic diversity determined in the step (2), wherein each population is most likely to collect rhizosphere soil and overground part orchid of all plants, placing the rhizosphere soil and plant materials in an ice box for storage, and bringing the rhizosphere soil and the plant materials back to a laboratory for treatment as soon as possible;
(4) placing the recovered rhizosphere soil in a sterilized conical flask, mixing the soil and water, adding a glucose solution, placing in a shaking table for culturing for 1-2h to obtain a soil suspension, and placing in a sterilized container;
(5) mixing the plants, grinding, placing in a sterilized conical flask, adding sterilized distilled water, adding glucose solution, culturing in a shaking table for 1-2 hr to obtain plant suspension, and placing in a sterilized container;
(6) standing the soil suspension in a sterilization container, and then mixing the soil suspension with sterilized common soil to prepare slurry;
(7) slightly scratching 8-11 scratches (with the length of 0.5cm and the depth of 2-3mm) on the stem of the hardened orchid tissue culture seedling by using a blade, soaking the scratched part in plant suspension in a sterilization container for 5-10min, and dipping the root of the tissue culture seedling in the slurry prepared in the step (6);
(8) and (4) planting or returning and nursing according to a conventional method.
Preferably, the glucose solution in step (4) and step (5) has a mass concentration of 5%, i.e. contains 5 g of glucose per 100 ml.
Preferably, the mixing of the soil and the water in the step (4) and the step (5) is carried out according to the ratio of soil: sterilized distilled water was added in a mass ratio of 1:20, and then a glucose solution was added in a ratio of 10% by weight.
Preferably, the soil suspension in step (6) is left standing for 10 min.
Compared with the prior art, the invention has the following beneficial effects:
(1) the population with the highest genetic diversity is selected through a molecular technology, generally speaking, the higher the genetic diversity of the population is, the stronger the adaptability of the population is, flora which can contribute to the self growth is formed in the plant and at the rhizosphere, and the adaptability and the resistance of the plants can be greatly improved by inoculating the flora into the tissue culture seedlings which do not form the flora;
(2) the method greatly improves the self resistance of the orchid, and further improves the transplanting survival rate of the wild orchid.
Detailed Description
The following detailed description is to be read in connection with specific embodiments, but it should be understood that the scope of the invention is not limited to the specific embodiments. The raw materials and reagents used in the examples were all commercially available unless otherwise specified. The rare endangered orchid variety adopted in the embodiment is the valuable cymbidium discoreanum.
Example 1
A method for improving the transplanting survival rate of rare or endangered orchid tissue culture seedlings specifically comprises the following operation steps:
(1) firstly, performing germplasm resource survey on rare species of orchid, collecting young leaves of 5-10 plants (two plants are more than 5 meters apart) from each colony, and storing in a sealed bag filled with silica gel;
(2) analyzing the genetic diversity of the transgenic plant by adopting molecular means such as ISSR, RAPD, simplified genome sequencing and the like, and determining the population with the highest genetic diversity;
(3) collecting rhizosphere soil and overground part orchid (not requiring the whole plant and having a length of 5-7 cm) of the population with the highest genetic diversity determined in the step (2), wherein each population is most likely to collect rhizosphere soil and overground part orchid of all plants, placing the rhizosphere soil and plant materials in an ice box for storage, and bringing the rhizosphere soil and the plant materials back to a laboratory for treatment as soon as possible;
(4) putting the recovered rhizosphere soil into a sterilized conical flask, and mixing the soil: adding sterilized distilled water in a mass ratio of 1:20, adding a glucose solution (the mass concentration of the glucose solution is 5%) in a ratio of 10% of the total weight, culturing in a shaking table for 1h to obtain a soil suspension, and placing in a sterilized container;
(5) mixing the returned plants, grinding, placing into a sterilized conical flask, adding sterilized distilled water, adding glucose solution (the mass concentration of the glucose solution is 5%) according to the total weight of 10%, placing into a shaking table, culturing for 1h to obtain plant suspension, and placing into a sterilized container;
(6) standing the soil suspension in a sterilization container for 10min, and mixing with sterilized common soil to obtain slurry;
(7) slightly scratching 10 scratches (with the length of 0.5cm and the depth of 2-3mm) on the stem of the hardened tissue culture seedling of the cymbidium goeringii by using a blade, soaking the scratched part in plant suspension in a sterilization container for 5-7min, and dipping the root of the tissue culture seedling in the slurry prepared in the step (6);
(8) the method is carried out according to a conventional method or regression conservation, the survival rate is counted after 2 months of field planting, and the survival rate is 96%.
Example 2
A method for improving the transplanting survival rate of rare or endangered orchid tissue culture seedlings specifically comprises the following operation steps:
(1) firstly, performing germplasm resource survey on rare species of orchid, collecting young leaves of 5-10 plants (two plants are more than 5 meters apart) from each colony, and storing in a sealed bag filled with silica gel;
(2) analyzing the genetic diversity of the transgenic plant by adopting molecular means such as ISSR, RAPD, simplified genome sequencing and the like, and determining the population with the highest genetic diversity;
(3) collecting rhizosphere soil and overground part orchid (not requiring the whole plant and having a length of 7-10 cm) of the population with the highest genetic diversity determined in the step (2), wherein each population is most likely to collect rhizosphere soil and overground part orchid of all plants, placing the rhizosphere soil and plant materials in an ice box for storage, and bringing the rhizosphere soil and the plant materials back to a laboratory for treatment as soon as possible;
(4) putting the recovered rhizosphere soil into a sterilized conical flask, and mixing the soil: adding sterilized distilled water in a mass ratio of 1:20, adding a glucose solution (the mass concentration of the glucose solution is 5%) in a ratio of 10% of the total weight, culturing in a shaking table for 2 hours to obtain a soil suspension, and placing in a sterilized container;
(5) mixing the returned plants, grinding, placing into a sterilized conical flask, adding sterilized distilled water, adding glucose solution (the mass concentration of the glucose solution is 5%) according to the total weight of 10%, placing into a shaking table, culturing for 2h to obtain plant suspension, and placing into a sterilized container;
(6) standing the soil suspension in a sterilization container for 10min, and mixing with sterilized common soil to obtain slurry;
(7) lightly scratching 9 scratches (with the length of 0.5cm and the depth of 2-3mm) on the stem of the hardened tissue culture seedling of the cymbidium goeringii by using a blade, soaking the scratched part in plant suspension in a sterilization container for 7-10min, and dipping the root of the tissue culture seedling in the slurry prepared in the step (6);
(8) the method is carried out according to a conventional method or regression conservation, the survival rate is counted after 2 months of field planting, and the survival rate is 97 percent.
The foregoing descriptions of specific exemplary embodiments of the present invention have been presented for purposes of illustration and description. It is not intended to limit the invention to the precise form disclosed, and obviously many modifications and variations are possible in light of the above teaching. The exemplary embodiments were chosen and described in order to explain certain principles of the invention and its practical application to enable one skilled in the art to make and use various exemplary embodiments of the invention and various alternatives and modifications as are suited to the particular use contemplated. It is intended that the scope of the invention be defined by the claims and their equivalents.
Claims (4)
1. A method for improving the transplanting survival rate of rare or endangered orchid tissue culture seedlings is characterized by comprising the following operation steps:
(1) firstly, performing germplasm resource survey on rare species of orchid, collecting young leaves of 5-10 plants for each colony, and storing;
(2) analyzing the genetic diversity of the plant by adopting ISSR, RAPD and simplified genome sequencing molecular means, and determining the population with the highest genetic diversity;
(3) collecting rhizosphere soil and overground part of orchid of the population with the highest genetic diversity determined in the step (2), and storing the rhizosphere soil and plant materials;
(4) placing the recovered rhizosphere soil in a bottle, mixing the soil and water, adding a glucose solution, culturing for 1-2h to obtain a soil suspension, and placing the soil suspension in a sterilized container;
(5) mixing, grinding and placing the plant with the seed back into a bottle, adding water, adding glucose solution, culturing for 1-2h to obtain plant suspension, and placing the plant suspension into a sterilized container;
(6) standing the soil suspension in a sterilization container, and then mixing the soil suspension with common soil to prepare slurry;
(7) 8-11 scratches are scribed on the stem of the hardened orchid tissue culture seedling, then the nicked part is soaked in the plant suspension for 5-10min, and then the root of the tissue culture seedling is dipped in the slurry prepared in the step (6);
(8) and (4) planting or returning and nursing according to a conventional method.
2. The method of claim 1, wherein: the mass concentration of the glucose solution in the step (4) and the step (5) is 5 percent.
3. The method of claim 1, wherein: in the step (4) and the step (5), the soil and the water are mixed according to the proportion of soil: water was added in a mass ratio of 1:20, and then a glucose solution was added in a ratio of 10% by weight.
4. The method of claim 1, wherein: and (5) standing the soil suspension in the step (6) for 10 min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111464725.2A CN114128585A (en) | 2021-12-03 | 2021-12-03 | Method for improving transplanting survival rate of rare or endangered orchid tissue culture seedlings |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202111464725.2A CN114128585A (en) | 2021-12-03 | 2021-12-03 | Method for improving transplanting survival rate of rare or endangered orchid tissue culture seedlings |
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| CN114128585A true CN114128585A (en) | 2022-03-04 |
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| CN202111464725.2A Pending CN114128585A (en) | 2021-12-03 | 2021-12-03 | Method for improving transplanting survival rate of rare or endangered orchid tissue culture seedlings |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5914107A (en) * | 1996-09-27 | 1999-06-22 | Mayekawa Manufacturing Co., Ltd. | Method of introducing an endophytic fungus into rough bluegrass belonging to Poa trivialis and Poa compressa |
| CN101307298A (en) * | 2008-01-30 | 2008-11-19 | 珠海市农业科学研究中心 | Broad-spectrum antifungal bacillus licheniformis and uses thereof |
| CN109757519A (en) * | 2018-12-27 | 2019-05-17 | 金华市胤宏农业科技有限公司 | A kind of rose transplantation rooting agent composition |
| CN111621517A (en) * | 2020-06-05 | 2020-09-04 | 杭州师范大学 | Method for inducing hairy roots of common turmeric |
-
2021
- 2021-12-03 CN CN202111464725.2A patent/CN114128585A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5914107A (en) * | 1996-09-27 | 1999-06-22 | Mayekawa Manufacturing Co., Ltd. | Method of introducing an endophytic fungus into rough bluegrass belonging to Poa trivialis and Poa compressa |
| CN101307298A (en) * | 2008-01-30 | 2008-11-19 | 珠海市农业科学研究中心 | Broad-spectrum antifungal bacillus licheniformis and uses thereof |
| CN109757519A (en) * | 2018-12-27 | 2019-05-17 | 金华市胤宏农业科技有限公司 | A kind of rose transplantation rooting agent composition |
| CN111621517A (en) * | 2020-06-05 | 2020-09-04 | 杭州师范大学 | Method for inducing hairy roots of common turmeric |
Non-Patent Citations (2)
| Title |
|---|
| 翟明恬等: "铁皮石斛组织快繁及移栽培养", 《东北林业大学学报》 * |
| 肖淑贤等: "植物内生菌的研究概况及应用进展", 《农业技术与装备》 * |
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