CN110771500A - Rapid propagation and breeding method of anoectochilus formosanus - Google Patents
Rapid propagation and breeding method of anoectochilus formosanus Download PDFInfo
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- CN110771500A CN110771500A CN201910956645.5A CN201910956645A CN110771500A CN 110771500 A CN110771500 A CN 110771500A CN 201910956645 A CN201910956645 A CN 201910956645A CN 110771500 A CN110771500 A CN 110771500A
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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
- A01H4/001—Culture apparatus for tissue culture
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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
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Abstract
The invention relates to a rapid propagation and breeding method of anoectochilus formosanus, which comprises the following steps: selecting wild anoectochilus formosanus with excellent properties, and picking full, sound and crack-free fruit pods after the pods are ripe; cleaning picked fruit pods, taking out seeds under aseptic condition, sterilizing the seeds on the surfaces, inoculating the seeds into a nutrient medium, placing the mixture in a constant temperature condition of 20-30 ℃, performing dark culture for 10-15 days, then placing the mixture in a sunlight shed with temperature and humidity control and 70% of light transmittance, performing culture for 90-120 days, observing the growth condition of the seeds during the culture, and removing pathological changes and polluted seedlings; hardening the seedlings for 3-5 days after the transplanting condition is reached, and transplanting; the breeding method of the invention has the advantages of high breeding speed, high rooting rate, high propagation coefficient and high transplanting survival rate, and can realize large-scale seedling.
Description
Technical Field
The invention relates to a rapid propagation and breeding method of anoectochilus formosanus, belonging to the technical field of planting.
Background
Anoectochilus formosanus (Anoectochilus formosanus)
Anoectochilus roxburghii) Is Orchidaceae (
Orchidaceae) The cheilot orchid is a perennial herb, also called anoectochilus formosanus, golden silkworm, anoectochilus formosanus and the like, and is mainly distributed in Fujian, Guangdong, Guangxi, Yunnan and other places in China. Anoectochilus roxburghii is a traditional Chinese precious medicinal material, is medicinal for the whole body, has mild taste and sweet nature, has the effects of clearing heat and cooling blood, removing dampness and detoxifying, prolonging life and the like, and can treat diseases such as lung heat cough, rheumatic bone pain, diabetes, cystitis and the like. The Anoectochilus roxburghii plant is small and attractive in shape and high in ornamental value, so that the Anoectochilus roxburghii plant has a wide application prospect. The anoectochilus roxburghii has tiny seeds, extremely low germination rate under natural conditions, short plants and slow growth and development, and animals such as insects, birds and the like to eat, so that the anoectochilus roxburghii becomes more rare. The cuttage propagation speed of anoectochilus formosanus is low. Therefore, the development of a rapid propagation and breeding method of anoectochilus formosanus can create conditions for protecting wild resources and reasonably developing and utilizing, and can also solve the problem of low survival rate and low propagation speed of the anoectochilus formosanus。
The invention provides a rapid propagation and breeding method of anoectochilus formosanus, which has the advantages of high propagation speed, high rooting rate, high propagation coefficient, high transplanting survival rate, capability of realizing large-scale seedling culture, capability of realizing simplified management and year reduction, and wide application value in production.
Disclosure of Invention
The invention relates to a rapid propagation and breeding method of anoectochilus formosanus, which comprises the following specific steps of selecting wild anoectochilus formosanus with excellent properties, and picking full, sound and crack-free fruit pods after pods are mature; cleaning picked fruit pods, taking out seeds under aseptic condition, sterilizing the seeds with a disinfectant, inoculating the seeds into a nutrient medium, placing the seeds in a constant temperature condition of 20-30 ℃, culturing in dark for 10-15 days, then placing the seeds in a sunlight shed with temperature and humidity control and 70% of light transmittance, culturing for 90-120 days, observing the growth condition of the seeds during the culturing period, and removing pathological changes and contaminated seedlings; hardening the seedlings for 3-5 days after the transplanting condition is reached, and transplanting; the breeding method of the invention has the advantages of high breeding speed, high rooting rate, high propagation coefficient and high transplanting survival rate, and can realize large-scale seedling.
In order to achieve the purpose, the invention adopts the technical scheme that:
a rapid propagation and breeding method of anoectochilus formosanus comprises the following steps:
(1) selecting wild anoectochilus formosanus with excellent properties, and picking full, sound and crack-free fruit pods as explants after the pods are mature;
(2) cleaning picked fruit pods, soaking seeds in 75% alcohol for 10 seconds under an aseptic condition, then putting in 10% sodium hypochlorite for disinfection for 15 minutes, cleaning for 6 times by using sterile water, draining, and putting in an aseptic culture dish for later use;
(3) inoculating the sterilized seeds into a nutrient medium, placing the nutrient medium under the constant temperature condition of 20-30 ℃, culturing in dark for 10-15 days, then placing the nutrient medium in a sunlight shed with temperature and humidity control and 70% of light transmittance, culturing for 90-120 days, observing the growth condition of the seeds in the period, and removing pathological changes and polluted seedlings;
(4) selecting seedling plants with height of 6-8cm, more than 3 leaves and 2-3 rooting, hardening seedlings for 3-5 days, transplanting, packaging and selling.
The nutrient medium formula comprises 21g/L of MS (Murashige & Skoog) as a basic culture medium, 2mg/L of 6-benzylaminopurine, 1.0-1.5mg/L of α -naphthylacetic acid, 0.05mg/L of brassinolide, 100g/L of banana, 6.5g/L of agar and 25g/L of sucrose.
The invention has the following advantages:
the breeding method of the invention has the advantages of high breeding speed, high rooting rate, high propagation coefficient and high transplanting survival rate, and can realize large-scale seedling.
Detailed Description
The present invention is further illustrated in detail by the following examples, which are provided only for illustrating the present invention and are not intended to limit the scope of the present invention.
Example 1
A rapid propagation and breeding method of anoectochilus formosanus comprises the following steps:
(1) selecting wild anoectochilus formosanus with excellent properties, and picking full, sound and crack-free fruit pods as explants after the pods are mature;
(2) cleaning picked fruit pods, soaking seeds in 75% alcohol for 10 seconds under an aseptic condition, then putting in 10% sodium hypochlorite for disinfection for 15 minutes, cleaning for 6 times by using sterile water, draining, and putting in an aseptic culture dish for later use;
(3) inoculating the sterilized seeds into a nutrient medium, placing the nutrient medium under the constant temperature condition of 25 ℃, culturing in the dark for 10 days, then placing the nutrient medium in a sunlight shed with temperature and humidity control and 70% of light transmittance, culturing for 120 days, observing the growth condition of the seeds in the sunlight shed, and removing pathological changes and polluted seedlings;
(4) selecting seedling plants with the height of 8cm, more than 3 leaves and 3 hairy roots, hardening the seedlings for 4 days, transplanting, packaging and selling.
The nutrient medium formula comprises 21g/L of MS (Murashige & Skoog) as a basic culture medium, 2mg/L of 6-benzylaminopurine, 1.4mg/L of α -naphthylacetic acid, 0.05mg/L of brassinolide, 100g/L of banana, 6.5g/L of agar and 25g/L of sucrose.
Example 2
A rapid propagation and breeding method of anoectochilus formosanus comprises the following steps:
(1) selecting wild anoectochilus formosanus with excellent properties, and picking full, sound and crack-free fruit pods as explants after the pods are mature;
(2) cleaning picked fruit pods, soaking seeds in 75% alcohol for 10 seconds under an aseptic condition, then putting in 10% sodium hypochlorite for disinfection for 15 minutes, cleaning for 6 times by using sterile water, draining, and putting in an aseptic culture dish for later use;
(3) inoculating the sterilized seeds into a nutrient medium, placing the nutrient medium under the constant temperature condition of 26 ℃, culturing for 12 days in a dark environment, then placing the nutrient medium in a sunlight shed with temperature and humidity control and 70% of light transmittance, culturing for 120 days, observing the growth condition of the seeds during the culturing period, and removing pathological changes and polluted seedlings;
(4) selecting seedling plants with the height of 6cm, more than 3 leaves and 2 hairy roots, hardening the seedlings for 4 days, transplanting, packaging and selling.
The nutrient medium formula comprises 21g/L of MS (Murashige & Skoog) as a basic culture medium, 2mg/L of 6-benzylaminopurine, 1.2mg/L of α -naphthylacetic acid, 0.05mg/L of brassinolide, 100g/L of banana, 6.5g/L of agar and 25g/L of sucrose.
Example 3
A rapid propagation and breeding method of anoectochilus formosanus comprises the following steps:
(1) selecting wild anoectochilus formosanus with excellent properties, and picking full, sound and crack-free fruit pods as explants after the pods are mature;
(2) cleaning picked fruit pods, soaking seeds in 75% alcohol for 10 seconds under an aseptic condition, then putting in 10% sodium hypochlorite for disinfection for 15 minutes, cleaning for 6 times by using sterile water, draining, and putting in an aseptic culture dish for later use;
(3) inoculating the sterilized seeds into a nutrient medium, placing the nutrient medium under the constant temperature condition of 20-30 ℃, culturing for 13 days in a dark environment, then placing the nutrient medium in a sunlight shed with temperature and humidity control and 70% of light transmittance, culturing for 100 days, observing the growth condition of the seeds in the dark environment, and removing pathological changes and contaminated seedlings;
(4) selecting seedling plants with height of 7cm, more than 3 leaves and 2 hairy roots, hardening seedlings for 3 days, transplanting, packaging and selling.
The nutrient medium formula comprises 21g/L of MS (Murashige & Skoog) as a basic culture medium, 2mg/L of 6-benzylaminopurine, 1.0mg/L of α -naphthylacetic acid, 0.05mg/L of brassinolide, 100g/L of banana, 6.5g/L of agar and 25g/L of sucrose.
Example 4
A rapid propagation and breeding method of anoectochilus formosanus comprises the following steps:
(1) selecting wild anoectochilus formosanus with excellent properties, and picking full, sound and crack-free fruit pods as explants after the pods are mature;
(2) cleaning picked fruit pods, soaking seeds in 75% alcohol for 10 seconds under an aseptic condition, then putting in 10% sodium hypochlorite for disinfection for 15 minutes, cleaning for 6 times by using sterile water, draining, and putting in an aseptic culture dish for later use;
(3) inoculating the sterilized seeds into a nutrient medium, placing the nutrient medium under the constant temperature condition of 30 ℃, culturing for 15 days in a dark environment, then placing the nutrient medium in a sunlight shed with temperature and humidity control and 70% of light transmittance, culturing for 120 days, observing the growth condition of the seeds during the culturing period, and removing pathological changes and polluted seedlings;
(4) selecting seedling plants with the height of 8cm, more than 3 leaves and 3 hairy roots, hardening the seedlings for 5 days, transplanting, packaging and selling.
The nutrient medium formula comprises 21g/L of MS (Murashige & Skoog) as a basic culture medium, 2mg/L of 6-benzylaminopurine, 1.5mg/L of α -naphthylacetic acid, 0.05mg/L of brassinolide, 100g/L of banana, 6.5g/L of agar and 25g/L of sucrose.
Claims (5)
1. A rapid propagation and breeding method of anoectochilus formosanus is characterized by comprising the following steps:
(1) selecting wild anoectochilus formosanus with excellent properties, and picking full, sound and crack-free fruit pods as explants after the pods are mature;
(2) cleaning picked fruit pods, taking out seeds under aseptic condition, and carrying out surface disinfection;
(3) inoculating the sterilized seeds into a nutrient medium, placing the nutrient medium under the constant temperature condition of 20-30 ℃, culturing in dark for 10-15 days, then placing the nutrient medium in a sunlight shed with temperature and humidity control and 70% of light transmittance, culturing for 90-120 days, observing the growth condition of the seeds in the period, and removing pathological changes and polluted seedlings;
(4) after the transplanting condition is met, hardening the seedlings for 3-5 days, transplanting, packaging and selling.
2. The rapid propagation and breeding method of anoectochilus formosanus according to claim 1, which is characterized in that: the specific method of the disinfection is that under the aseptic condition, the seeds are soaked in 75% alcohol for 10 seconds, then the seeds are disinfected in 10% sodium hypochlorite for 15 minutes, washed for 6 times by sterile water, drained and placed in an aseptic culture dish for later use.
3. The method for rapid propagation and breeding of Anoectochilus roxburghii according to claim 1, wherein the nutrient medium formula comprises 21g/L of MS as a basic culture medium, 2mg/L of 6-benzylaminopurine, 1.0-1.5mg/L of α -naphthylacetic acid, 0.05mg/L of brassinolide, 100g/L of banana, 6.5g/L of agar and 25g/L of sucrose.
4. The rapid propagation and breeding method of anoectochilus formosanus according to claim 1, which is characterized in that: the nutrient medium is prepared by mixing the above components in the nutrient medium formulation of claim 3, heating until the agar is completely dissolved, and adjusting pH to 5.6.
5. The rapid propagation and breeding method of anoectochilus formosanus according to claim 1, which is characterized in that: the transplanting conditions are that the height of the seedling is 6-8cm, more than 3 leaves are adopted, and 2-3 roots are generated.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101213942A (en) * | 2008-01-10 | 2008-07-09 | 浙江省中药研究所有限公司 | Medicinal anoectochilus Formosan tissue culture one-step seedling establishment fast replication method |
| CN104823847A (en) * | 2015-04-29 | 2015-08-12 | 福建省农业科学院农业生物资源研究所 | Zhejiang anoectochilus roxburghii seed tissue culture and rapid seedling raising method |
| CN107372067A (en) * | 2017-08-16 | 2017-11-24 | 广东中科农业科技有限公司 | A kind of method for cultivating seedlings of anoectochilus |
| CN107743868A (en) * | 2017-10-29 | 2018-03-02 | 广西壮族自治区中国科学院广西植物研究所 | A kind of method for efficiently breeding roxburgh anoectochilus terminal bud using nature optical culture forming seedling through one step culture |
-
2019
- 2019-10-10 CN CN201910956645.5A patent/CN110771500A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101213942A (en) * | 2008-01-10 | 2008-07-09 | 浙江省中药研究所有限公司 | Medicinal anoectochilus Formosan tissue culture one-step seedling establishment fast replication method |
| CN104823847A (en) * | 2015-04-29 | 2015-08-12 | 福建省农业科学院农业生物资源研究所 | Zhejiang anoectochilus roxburghii seed tissue culture and rapid seedling raising method |
| CN107372067A (en) * | 2017-08-16 | 2017-11-24 | 广东中科农业科技有限公司 | A kind of method for cultivating seedlings of anoectochilus |
| CN107743868A (en) * | 2017-10-29 | 2018-03-02 | 广西壮族自治区中国科学院广西植物研究所 | A kind of method for efficiently breeding roxburgh anoectochilus terminal bud using nature optical culture forming seedling through one step culture |
Non-Patent Citations (3)
| Title |
|---|
| 付传明等: "广西金线莲种源特性及无菌播种萌发的影响因素", 《分子植物育种》 * |
| 何碧珠等: "多倍体金线莲一次性成苗及移栽", 《福建农林大学学报(自然科学版)》 * |
| 陈园媛等: "金线莲一次性成苗技术研究", 《江苏农业科学》 * |
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Application publication date: 20200211 |