CN110972956A - Pleione seed tissue culture method - Google Patents
Pleione seed tissue culture method Download PDFInfo
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- CN110972956A CN110972956A CN202010001874.4A CN202010001874A CN110972956A CN 110972956 A CN110972956 A CN 110972956A CN 202010001874 A CN202010001874 A CN 202010001874A CN 110972956 A CN110972956 A CN 110972956A
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- China
- Prior art keywords
- pleione
- seeds
- tissue culture
- culture dish
- bulbocodioides
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- 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
Abstract
The invention relates to the technical field of Pleione planting, and discloses a Pleione seed tissue culture method, which comprises the following steps: the harvested Pleione seeds were removed of the undesired parts, the desired parts were carefully washed clean, and then the seeds were cut to the appropriate size. According to the Pleione seed tissue culture method, tissue culture is performed, large-scale tissue culture can be performed on Pleione in a large-scale Pleione planting base, so that the seedling survival rate of Pleione is increased, the Pleione yield is guaranteed, the reproduced progeny is uniform, the excellent properties of the original variety can be maintained, and the Pleione seed proliferation and rhizome culture processes can not be rotten or necrotic after sterilization.
Description
Technical Field
The invention relates to the technical field of Pleione planting, in particular to a Pleione seed tissue culture method.
Background
The concept of plant tissue culture, also called isolated culture, refers to a technique of separating desired tissues, organs or cells, protoplasts, etc. from a plant body, and culturing under an artificial control condition by aseptic manipulation to obtain a regenerated whole plant or produce other products having economic value.
According to the culture medium for tissue culture of the plants of the genus Euphorbia and the method thereof provided by the invention authorized in China CN 201710183796.2, the culture medium provided by the invention has the beneficial effects that the propagation coefficient of the plants of the genus Euphorbia can be obviously improved, the propagation period is shortened, and a more effective way is provided for industrial production of the plants of the genus Euphorbia, but the traditional Pleione seed has a lower survival rate, and can carry a large amount of subsequent germs to affect the growth of the Pleione, and if the planting method provided by the invention is adopted, the disinfection effect is poorer, the survival rate of the Pleione seed is difficult to improve, so the Pleione seed tissue culture method is provided.
Disclosure of Invention
Technical problem to be solved
Aiming at the defects of the prior art, the Pleione seed tissue culture method provided by the invention has the advantages of high survival rate, good sterilization effect and the like, and solves the problems that the traditional Pleione seed has low survival rate, can carry a large amount of subsequent germs to influence the growth of Pleione, and if the planting method provided by the invention is adopted, the sterilization effect is poor, and the survival rate of Pleione seed is difficult to improve.
(II) technical scheme
In order to achieve the purposes of high survival rate and good sterilization effect, the invention provides the following technical scheme: a tissue culture method of Pleione bulbocodioides seeds comprises the following steps:
1) removing unnecessary parts of the collected Pleione seeds, carefully cleaning the required parts, cutting the seeds into proper sizes, placing the Pleione seeds in a gauze bag, placing the gauze bag in a faucet, and flushing for 10-40 minutes;
2) preparing a disinfected beaker, a glass tube, 70% alcohol, disinfectant, sterile water, a watch and the like, soaking the treated Pleione bulbocodioides in the step 1) for 5-10 minutes by using 70% alcohol, taking out the Pleione bulbocodioides, and peeling off the skin of the Pleione bulbocodioides after the alcohol is evaporated;
3) soaking the disinfected Pleione seeds in a sterilizing agent, taking out the seeds, and brushing the seeds with sterile water for 3 minutes each time and repeatedly brushing for 5-10 times;
4) the disinfected pleione seed is subjected to aseptic peeling by using tools such as a sterile knife, a shear, a forceps and the like under the aseptic condition;
5) under the aseptic environment, the cut Pleione seeds are immediately connected to a culture dish, 1-2 seeds are inoculated to each bottle, the culture dish is sealed by using an aseptic adhesive tape, and the culture dish is placed in a space of 25 ℃;
6) the propagation coefficient of the Pleione seeds needs to be enlarged after the Pleione seeds grow new shoots, at the moment, the new shoots are subjected to division or segmentation by adopting sterile scissors, then the divided Pleione seed new shoots are transferred into a proliferation culture dish, a nutrient solution is added into the culture dish, and proliferation can be continued according to the situation after one month of proliferation;
7) transferring the seed of the Pleione after sprouting into a rooting culture dish for rooting, after 1 month, the Pleione will grow the rhizome, at the moment, opening the culture dish, letting the culture dish to be under natural illumination for 2-3 days, taking out the Pleione plantlet, washing the nutrient solution on the root system of the plantlet with tap water, and then transplanting the Pleione nursery into a planting apparatus.
Preferably, the Pleione seeds are full, free of wound and free of diseases and pests, and the Pleione seeds are washed with a brush during cleaning.
Preferably, the size of the cut Pleione seeds is adapted to the size of the culture dish, and the gauze bag used for washing is sterilized in advance.
Preferably, when the Pleione seeds are washed, a cleaning solution can be added for cleaning, and then the cleaning solution is washed clean, and the Pleione seeds can be stirred when being sterilized and soaked.
Preferably, the petri dish is sterilized before use, dried by high temperature and placed in a sterile environment.
Preferably, the Pleione bulbocodioides bud is cut into small pieces with equal distance when being proliferated, and the process is completed in a sterile environment, and the nutrient solution is prepared by mixing nitrate nitrogen, phosphate, sulfate, potassium, sodium and chloride.
(III) advantageous effects
Compared with the prior art, the invention provides a Pleione seed tissue culture method, which has the following beneficial effects:
1. the Pleione seed tissue culture method can propagate Pleione rapidly in a short time depending on natural conditions through tissue culture, and disinfects Pleione seeds by adopting multiple disinfectants or disinfection methods during culture, thereby reducing the influence of germs on the growth of Pleione seeds, allowing Pleione seeds to grow under aseptic conditions, keeping the time in an aseptic environment, reducing the influence of germs on Pleione seeds, and performing large-scale tissue culture on Pleione in a large-scale Pleione planting base, thereby increasing the survival rate of Pleione seedlings, ensuring the excellent rate of Pleione, and keeping the good characters of original varieties.
2. According to the Pleione seed tissue culture method, by means of various disinfection methods, in the Pleione seed tissue culture process, the methods of washing, alcohol soaking, sterilizing by an aesthetic sterilizing agent and operating in a sterile environment are adopted, the Pleione seeds are sufficiently sterilized, the sterilization effect of the Pleione culture method is effectively improved, so that the survival rate of the Pleione seeds is increased, and the Pleione seeds are not rotten or necrosed in the Pleione seed proliferation and rhizome culture processes after sterilization.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Experimental example: a tissue culture method of Pleione bulbocodioides seeds comprises the following steps:
1) removing unnecessary parts of the harvested Pleione seeds, carefully cleaning the required parts, cutting the seeds into proper sizes, wherein the sizes of the cut Pleione seeds are matched with the sizes of culture dishes, a gauze bag used for washing is sterilized in advance, then the Pleione seeds are placed in the gauze bag and are flushed by running water from a faucet for 20 minutes, the Pleione seeds are selected from full, wound-free and pest-free survival-period seeds, the Pleione seeds are washed by a brush when being washed, a cleaning solution can be added for washing when the Pleione seeds are washed, and then the cleaning solution is washed clean, and the Pleione seeds can be stirred when being sterilized and soaked;
2) preparing a disinfected beaker, a glass tube, 70% alcohol, disinfectant, sterile water, a watch and the like, soaking the treated Pleione bulbocodioides in the step 1) for 6 minutes by using the 70% alcohol, taking out the Pleione bulbocodioides, and peeling off the skins of the Pleione bulbocodioides after the alcohol is evaporated;
3) soaking the disinfected Pleione seeds in a sterilizing agent, taking out the seeds, and brushing the seeds with sterile water for 3 minutes each time and repeatedly brushing for 6 times;
4) the disinfected pleione seed is subjected to aseptic peeling by using tools such as a sterile knife, a shear, a forceps and the like under the aseptic condition;
5) under the aseptic environment, the cut Pleione seeds are immediately connected to a culture dish, 2 seeds are inoculated in each bottle, the culture dish is sealed by using an aseptic tape and is placed in a space of 25 ℃, the culture dish needs to be sterilized before use, and then is dried at high temperature and placed under the aseptic environment;
6) the propagation coefficient of the Pleione seeds needs to be enlarged after the Pleione seeds grow new shoots, at the moment, the new shoots are subjected to division or segmentation by adopting sterile scissors, then the divided Pleione seed new shoots are transferred into a proliferation culture dish, a nutrient solution is added into the culture dish, proliferation can be continued according to the situation after one month of proliferation, the Pleione new shoots need to be cut into small blocks with equal distance when being subjected to proliferation, the process is finished in a sterile environment, and the nutrient solution is prepared by mixing nitrate nitrogen, phosphate, sulfate, potassium, sodium and chloride;
7) transferring the seed of the Pleione after sprouting into a rooting culture dish for rooting, after 1 month, the Pleione will grow the rhizome, at the moment, opening the culture dish, letting the culture dish stand under natural light for 2 days, then taking out the Pleione plantlet, washing the nutrient solution on the root system of the plantlet with tap water, and then transplanting the Pleione nursery into a planting apparatus.
Experiments prove that the survival rate of the Pleione can be effectively increased by cultivating the Pleione by adopting a tissue cultivation method, the bred offspring is uniform, the excellent properties of the original variety can be maintained, the plant has strong growth vigor, enlarged flowers, bright color, improved stress resistance and increased flower yield, the growth and differentiation rules of the cultivated part can be researched without being interfered by other parts of the plant body, the growth and differentiation of the cultivated part can be influenced by various cultivation conditions, the problems in theory and production can be solved, the development and mutual infiltration of plant physiology, biochemistry, genetics, cytology and morphology, agriculture, forestry, medicine and other subjects in the biological science can be powerfully promoted, and the research on nutrition physiology, cell physiology and metabolism, biosynthesis, gene transfer and gene recombination can be promoted.
The invention has the beneficial effects that: the tissue culture method of the Pleione seeds can quickly propagate the Pleione in a short time depending on natural conditions through tissue culture, and the Pleione seeds are sterilized by adopting a plurality of disinfectants or sterilization methods during the culture, so that the influence of germs on the growth of the Pleione seeds is reduced, the Pleione seeds are grown under the aseptic condition, the time is in the aseptic environment, the influence of the germs on the Pleione seeds is reduced, the Pleione can be subjected to large-scale tissue culture in a large-scale Pleione planting base, the seedling survival rate of the Pleione is increased, the excellent yield of the Pleione is ensured, the propagated offspring is uniform, the excellent characters of the original variety can be kept, and the tissue culture process of the Pleione seeds adopts the methods of washing, alcohol soaking, sterilizing by an aesthetic sterilizing agent and operating in the aseptic environment through a plurality of sterilization methods, the Pleione seeds are sufficiently sterilized, so that the sterilization effect of the Pleione cultivation method is effectively improved, the survival rate of the Pleione seeds is increased, and the Pleione seeds are not rotten or necrosed in the processes of proliferation and rhizome culture after sterilization.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (6)
1. The tissue culture method of the Pleione seeds is characterized by comprising the following steps:
1) removing unnecessary parts of the collected Pleione seeds, carefully cleaning the required parts, cutting the seeds into proper sizes, placing the Pleione seeds in a gauze bag, placing the gauze bag in a faucet, and flushing for 10-40 minutes;
2) preparing a disinfected beaker, a glass tube, 70% alcohol, disinfectant, sterile water, a watch and the like, soaking the treated Pleione bulbocodioides in the step 1) for 5-10 minutes by using 70% alcohol, taking out the Pleione bulbocodioides, and peeling off the skin of the Pleione bulbocodioides after the alcohol is evaporated;
3) soaking the disinfected Pleione seeds in a sterilizing agent, taking out the seeds, and brushing the seeds with sterile water for 3 minutes each time and repeatedly brushing for 5-10 times;
4) the disinfected pleione seed is subjected to aseptic peeling by using tools such as a sterile knife, a shear, a forceps and the like under the aseptic condition;
5) under the aseptic environment, the cut Pleione seeds are immediately connected to a culture dish, 1-2 seeds are inoculated to each bottle, the culture dish is sealed by using an aseptic adhesive tape, and the culture dish is placed in a space of 25 ℃;
6) the propagation coefficient of the Pleione seeds needs to be enlarged after the Pleione seeds grow new shoots, at the moment, the new shoots are subjected to division or segmentation by adopting sterile scissors, then the divided Pleione seed new shoots are transferred into a proliferation culture dish, a nutrient solution is added into the culture dish, and proliferation can be continued according to the situation after one month of proliferation;
7) transferring the seed of the Pleione after sprouting into a rooting culture dish for rooting, after 1 month, the Pleione will grow the rhizome, at the moment, opening the culture dish, letting the culture dish to be under natural illumination for 2-3 days, taking out the Pleione plantlet, washing the nutrient solution on the root system of the plantlet with tap water, and then transplanting the Pleione nursery into a planting apparatus.
2. The tissue culture method of Pleione bulbocodioides seeds of claim 1, wherein: the Pleione seeds are full, free of wound and disease and insect, and are cleaned by a brush when being cleaned.
3. The tissue culture method of Pleione bulbocodioides seeds of claim 1, wherein: the size of the cut Pleione seeds is adapted to the size of the culture dish, and the gauze bag used for washing should be sterilized in advance.
4. The tissue culture method of Pleione bulbocodioides seeds of claim 1, wherein: the pleione seed washing device is characterized in that a cleaning solution can be added to wash the pleione seeds when the pleione seeds are washed, and then the pleione seeds can be washed clean, and the pleione seeds can be stirred when the pleione seeds are sterilized and soaked.
5. The tissue culture method of Pleione bulbocodioides seeds of claim 1, wherein: the petri dish needs to be sterilized before use, then dried by high temperature and placed in a sterile environment.
6. The tissue culture method of Pleione bulbocodioides seeds of claim 1, wherein: the Pleione bulbocodioides bud is cut into small pieces with equal distance when being proliferated, the process is finished in a sterile environment, and the nutrient solution is prepared by mixing nitrate nitrogen, phosphate, sulfate, potassium, sodium and chloride.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202010001874.4A CN110972956A (en) | 2020-01-02 | 2020-01-02 | Pleione seed tissue culture method |
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| CN202010001874.4A CN110972956A (en) | 2020-01-02 | 2020-01-02 | Pleione seed tissue culture method |
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| CN110972956A true CN110972956A (en) | 2020-04-10 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114208681A (en) * | 2022-01-24 | 2022-03-22 | 中国科学院昆明植物研究所 | Energy-saving simple Pleione seedling culture method |
| CN114958607A (en) * | 2022-06-22 | 2022-08-30 | 肖尊平 | Compound microbial agent with high preparation efficiency and preparation method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007004390A1 (en) * | 2005-07-06 | 2007-01-11 | Kenji Kohno | Cachexia improving preparation and cachexia improving food |
| CN100998311A (en) * | 2007-01-04 | 2007-07-18 | 西南交通大学 | Method for artificially breeding edulis mono-garlic clove orchid |
| CN106332782A (en) * | 2016-10-19 | 2017-01-18 | 贵州省农作物品种资源研究所 | Single-step seedling culture method for pleione bulbocodioides through tissue culture |
| CN108012929A (en) * | 2017-12-27 | 2018-05-11 | 康美药业（文山）药材种植管理有限公司 | A kind of tissue cultures expanding propagation method of pleione bulbocodioides |
-
2020
- 2020-01-02 CN CN202010001874.4A patent/CN110972956A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007004390A1 (en) * | 2005-07-06 | 2007-01-11 | Kenji Kohno | Cachexia improving preparation and cachexia improving food |
| CN100998311A (en) * | 2007-01-04 | 2007-07-18 | 西南交通大学 | Method for artificially breeding edulis mono-garlic clove orchid |
| CN106332782A (en) * | 2016-10-19 | 2017-01-18 | 贵州省农作物品种资源研究所 | Single-step seedling culture method for pleione bulbocodioides through tissue culture |
| CN108012929A (en) * | 2017-12-27 | 2018-05-11 | 康美药业（文山）药材种植管理有限公司 | A kind of tissue cultures expanding propagation method of pleione bulbocodioides |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114208681A (en) * | 2022-01-24 | 2022-03-22 | 中国科学院昆明植物研究所 | Energy-saving simple Pleione seedling culture method |
| CN114958607A (en) * | 2022-06-22 | 2022-08-30 | 肖尊平 | Compound microbial agent with high preparation efficiency and preparation method thereof |
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