CN107318556B - In-vitro cultivation of elm longissima fruits and sowing and seedling method thereof - Google Patents
In-vitro cultivation of elm longissima fruits and sowing and seedling method thereof Download PDFInfo
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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
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- 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
- A01G17/00—Cultivation of hops, vines, fruit trees, or like trees
- A01G17/005—Cultivation methods
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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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Abstract
An in-vitro cultivation and sowing and seedling method of elm longissima fruits belongs to the technical field of plant breeding. The method mainly comprises the steps of mother tree selection, branch collection, in-vitro cultivation, fruit harvesting, sowing, seedling management, transplanting, potting and the like. The application of the method can easily collect and cultivate the fruits and seeds of elm ulmoides, solve the problem that the fruits and seeds of elm ulmoides are difficult to collect manually, effectively ensure the quality of elm seeds, and improve the germination rate and seedling rate of sowing and seedling.
Description
Technical Field
The invention belongs to the technical field of plant breeding, and particularly relates to an artificial breeding method of ulmus pumila.
Background
Ulmus pumila (Ulmus elsongata) is a new species of Ulmus pumila published by 20 th century 80 th-school botanicals in the beginning and middle schools according to the specimens of Zhejiang tunneling, has excellent material and is an excellent fast-growing wood tree species with a promising future. The classification of Ulmus Davidiana Hance belongs to Ulmus Davidiana Hance (Chaetopelea) of Ulmus of Ulmaceae (Ulmaceae), only 4 species exist in the world, 3 species are produced in North America and 1 species in China, and are distributed sporadically only in Zhejiang, Fujian, Anhui and Jiangxi. The few or imminent extinction is listed as a country
The grade emphasis protects the plants. The main reasons for endangering extinction of ulmus pumila are as follows: one is as follows: the samara is thin and light, and when the samara is mature, the samara will drift with the wind, and the manual collection of the elm seeds is difficult; the second step is as follows: under the condition of natural environment, the effective propagation distance of the fruits is short, and the survival rate of the naturally planted fruits is low due to the discomfort of the habitat (such as illumination, temperature, humidity and the like), so that the propagation is difficult, seedlings are lacked in the community, the updating is blocked, and the population is in a decline trend. The elm of long-ordered origin is in the edge of endangering extinction, and needs to be artificially protected.
Disclosure of Invention
In order to solve the defects in the prior art, the invention provides a method for in vitro cultivation of elm longissima fruits and sowing and seedling.
The invention adopts the following technical scheme:
the in vitro cultivation method of the elm longituba fruits is characterized by mainly comprising the following steps:
1) selecting a mother tree: selecting a strong, straight and disease and insect pest-free mother tree, and planting the mother tree for 20-30 years;
2) branch collection: when the flower buds of elm occur but the inflorescence is not formed, high-branch scissors are used for shearing branches with the flower buds, the branches are taken back to a laboratory, and the flower buds on the branch sections are required to be full and healthy;
3) in vitro cultivation: cutting the collected branch sections into branch sections with the length of 30-40 cm, inserting the branch sections into a container bottle for liquid cultivation, and keeping the environmental temperature at 10-20 ℃ and the humidity at 70-80%; replacing the culture solution every 5-7 days; preferably, the liquid culture is nutrient solution culture, and the formula of the nutrient solution is as follows: potassium nitrate 607 mg/L, calcium nitrate 945 mg/L, ammonium phosphate 115 mg/L, magnesium sulfate 493 mg/L, GGR 6 No. 50 mg/L, aspirin 100 mg/L, activated carbon 2 mg/L, iron salt solution 2.5 ml/L, trace element solution 5ml/L, pH = 6.0; the iron salt solution comprises the following formula: 2.78 g of ferrous sulfate heptahydrate, 3.73 g of ethylenediamine disodium acetate and 500 ml of distilled water; the formula of the trace element solution is as follows: potassium iodide 0.83mg/L, boric acid 6.2 mg/L, manganese sulfate 22.3 mg/L, zinc sulfate 8.6 mg/L, sodium molybdate 0.25 mg/L, copper sulfate 0.025mg/L, cobalt chloride 0.025 mg/L;
4) harvesting fruits: when the samara turns yellow-brown to ripe from green, the samara is collected in time.
The sowing and seedling raising method for elms in the long-ordered market is characterized by comprising the following steps:
1) sowing: removing impurities from the ulmus davidiana fruits according to claim 1 or 2, performing plug seeding, seedling raising and seeding within 2-3 days after the fruits are harvested, watering thoroughly after seeding, and keeping the temperature of a matrix at 20-25 ℃; preferably, the seedling substrate is any one of river sand, vegetable garden soil or special seedling peat; the special seedling peat is No. 1 German Viter seedling peat produced by Hangzhou rainbow flower Co Ltd;
2) seedling stage management: mainly comprises temperature management, water and fertilizer management, illumination control and pest control; preferably, the method comprises the following steps: the temperature management is as follows: keeping the temperature at about 25 ℃ in the day and preferably at 16-20 ℃ at night but not lower than 10 ℃ before the seedlings come out of the earth and cotyledons are completely unfolded, and adjusting the temperature by shading, ventilating and spraying water; the water and fertilizer management method comprises the following steps: spraying small water with a spraying pot after seedling emergence, wetting the substrates in a dry and wet mode, spraying foliar fertilizer for about 4 times by combining water spraying, and alternately spraying water-soluble fertilizers 20-10-20 and 14-0-14 at the concentration of 50-100 mg/kg for 7 days; the illumination control method comprises the following steps: after the seedlings come out of the soil, the illumination intensity and the illumination hours are gradually increased, wherein the illumination intensity is 1500-2500 Lux, and the illumination hours are 8-10 h/d. The pest control method comprises the following steps: after the seedlings come out of the soil, alternately spraying 800 times of carbendazim or thiophanate methyl for 7-10 days on average once, and frequently ventilating a seedbed;
3) transplanting and potting: and (5) transplanting the seedlings into a pot in time when 3-4 true leaves grow out of the seedlings.
The in vitro cultivation and sowing seedling method of the elm fruit provides an artificial seed collecting and seedling method, overcomes the technical defect of difficult natural propagation, solves the problem of difficult artificial collection of elm fruit and seeds, ensures the quality of elm seeds, and increases the seed bearing rate of each fruit by 86.0 percent and 21.7 percent compared with the cultivation of clear water through the elm fruit branches cultivated in vitro by nutrient solution; the special seedling peat is adopted for plug-seedling, the average germination rate reaches 94.2 percent, and is respectively improved by 5.7 percent and 15.1 percent compared with river sand and common vegetable garden soil; the special peat for seedling culture is adopted for seedling culture, the average seedling rate reaches 94.2 percent, and is respectively improved by 18.8 percent and 15.9 percent compared with river sand and common vegetable garden soil. The invention provides a whole set of elm ulmus davidiana fruit in-vitro cultivation and sowing and seedling method thereof, which comprises mother tree selection, branch collection time, nutrient solution preparation, in-vitro cultivation, seed collection, sowing and seedling, ensures the quality of elm ulmus davidiana fruit seeds, improves the germination rate and seedling rate of elm ulmus davidiana seeds, and has strong promotion effect on urban garden application and popularization of excellent local trees and forestry ecological construction.
Detailed Description
Example 1.
1. Selecting a mother tree: selecting a strong, straight and disease and insect pest-free mother tree, and planting the mother tree for 20 years.
2. And (3) strip picking time: in the last 3 months of 2016, in sunny days, when flower buds appear but inflorescences are not formed, branches with the flower buds are cut by using a tree pruner and taken back to a laboratory, and the flower buds on the branch segments are required to be full and healthy.
3. Preparing a nutrient solution: the following raw materials are weighed according to the weight, added into water in sequence and stirred continuously to be fully dissolved and mixed into the required nutrient solution. Potassium nitrate 607 mg/L, calcium nitrate 945 mg/L, ammonium phosphate 115 mg/L, magnesium sulfate 493 mg/L, GGR 6 No. 50 mg/L, aspirin 100 mg/L, activated carbon 2 mg/L, iron salt solution 2.5 ml/L (ferrous sulfate heptahydrate 2.78 g, disodium ethylenediamine acetate 3.73 g, distilled water 500 ml, pH = 5.5), trace elements 5ml/L (potassium iodide 0.83mg/L, boric acid 6.2 mg/L, manganese sulfate 22.3 mg/L, zinc sulfate 8.6 mg/L, sodium molybdate 0.25 mg/L, copper sulfate 0.025mg/L, cobalt chloride 0.025 mg/L), pH = 6.0.
4. In vitro cultivation: the prepared nutrient solution is subpackaged in glass bottles, collected branch sections are cut into branch sections with the length of 30-40 cm, the branch sections are inserted into container bottles for cultivation, the culture solution is replaced every 5-7 days, the culture solution is cultivated with clear water as a control, the environment temperature is kept at 10-20 ℃, the humidity is kept at 70-80%, and the test results are shown in table 1.
TABLE 1 Effect of different nutrient solution treatments on the fruiting quantity and fruit growth and development of Ulmus Davidiana Hance
| Test treatment | Quantity of results (grain/branch) | Real number of ripe fruit (grain) | Fruit seed setting rate% |
| Clear water control | 28 | 19 | 67.9 |
| Nutrient solution | 29 | 25 | 86.2 |
5. Harvesting fruits: and after 18-22 days, harvesting in time when the samara turns yellow-brown to mature from green.
6. Sowing and seedling raising: and after the fruits are harvested, removing impurities, and performing plug seeding, seedling raising and seeding within 2-3 days after the fruits are harvested. And (2) respectively adopting 3 matrixes of river sand, vegetable garden soil and special seedling peat (the special seedling peat is purchased from Hangzhou Hongyou flower Co., Ltd., German Viter seedling No. 1 peat) to perform plug seeding and seedling culture, wherein in the plug seeding process, one plug is used for plug seeding, water is permeated after seeding, and the temperature of the matrix is kept at 20-25 ℃. And performing conventional seedling raising treatment in the later stage, keeping the substrate wet, and germinating 3-5 days after sowing. The test results are shown in Table 2.
TABLE 2 Effect of different media on the germination and seedling development of Ulmus Davidiana Hance seeds
| Class of substrate | Time of sowing | Budding time (d) | The germination percentage is% | The seedling rate% |
| River sand | 4 month and 9 days | 4 month and 12 days | 90.2 | 70.4 |
| Common vegetable garden soil | 4 month and 9 days | 4 month and 15 days | 80.4 | 80.0 |
| Special peat for seedling culture | 4 month and 9 days | 4 month and 15 days | 94.1 | 93.0 |
7. Seedling stage management: (1) temperature management: the temperature is kept at about 25 ℃ in the day and preferably at 16-20 ℃ at night but not lower than 10 ℃ before the seedlings come out of the earth and cotyledons are completely unfolded, and the temperature can be adjusted by shading, ventilating, spraying water and the like. (2) And (3) water and fertilizer management: spraying small water with a spraying pot after seedling emergence, wetting the substrates, spraying foliar fertilizer for about 4 times by combining water spraying, and spraying 20-10-20 and 14-0-14 water-soluble fertilizers (purchased from Hangzhou Hongyou flower Co., Ltd.; 20-10-20 comprises 20% of N, 10% of P and 20% of K, 14-0-14 comprises 14% of N, 0% of P and 14% of K alternately for 7 days at a concentration of 50-100 mg/kg. (3) And (3) illumination control: the illumination intensity is 1500-2500 Lux, and the illumination hours are 8-10 h/d; when the seedlings come out of the earth, gradually increasing the illumination intensity and the illumination hours of the seedbed by replacing a shading net or turning on a light supplement lamp, wherein the illumination intensity is increased from 1500Lux to 2500Lux, and the illumination hours are increased from 8h/d to 10 h/d; increasing 500Lux illumination intensity and 1h/d illumination hours every 7-10 days, wherein the maximum illumination intensity in a seedling stage is 2500Lux, and the maximum illumination hours is 10 h/d. (4) And (3) pest control: the prevention is mainly carried out, after the seedlings come out of the soil, 800 times of liquid carbendazim or thiophanate methyl is sprayed for 7-10 days on average alternately, and the seedbed is often ventilated.
8. Transplanting and potting: and (3) transplanting the seedlings into a pot in time when the seedlings grow 3-4 true leaves (the height reaches 2-6 cm).
Example 2.
1. Selecting a mother tree: selecting a strong, straight and disease and insect pest-free mother tree, and planting the mother tree for 30 years.
2. And (3) strip picking time: in the last 4 th month of 2017, in windless sunny days, when flower buds appear but inflorescences are not formed, branches with the flower buds are cut by using a tree pruner and taken back to a laboratory, and the flower buds on the branch segments are required to be full and healthy.
3. Preparing a nutrient solution: the following raw materials are weighed according to the weight, added into water in sequence and stirred continuously to be fully dissolved and mixed into the required nutrient solution. Potassium nitrate 607 mg/L, calcium nitrate 945 mg/L, ammonium phosphate 115 mg/L, magnesium sulfate 493 mg/L, GGR 6 No. 50 mg/L, aspirin 100 mg/L, activated carbon 2 mg/L, iron salt solution 2.5 ml/L (ferrous sulfate heptahydrate 2.78 g, disodium ethylenediamine acetate 3.73 g, distilled water 500 ml, pH = 5.5), trace elements 5ml/L (potassium iodide 0.83mg/L, boric acid 6.2 mg/L, manganese sulfate 22.3 mg/L, zinc sulfate 8.6 mg/L, sodium molybdate 0.25 mg/L, copper sulfate 0.025mg/L, cobalt chloride 0.025 mg/L), pH = 6.0.
4. In vitro cultivation: the prepared nutrient solution is subpackaged in container bottles (glass bottles or mineral water bottles), then collected branch sections are cut into branch sections with the length of 30-40 cm, the branch sections are inserted into the container bottles for cultivation, the culture solution is replaced every 5-7 days, the cultivation is carried out by using clear water as a reference, the environmental temperature is kept at 10-20 ℃, the humidity is kept at 70-80%, and the test results are shown in table 3.
TABLE 3 Effect of different nutrient solution treatments on the fruiting quantity and fruit growth and development of Ulmus Davidiana Hance
| Test treatment | Quantity of results (grain/branch) | Real number of ripe fruit (grain) | Fruit seed setting rate% |
| Clear water control | 33 | 20 | 60.6 |
| Nutrient solution | 35 | 30 | 85.7 |
5. Harvesting fruits: and after 18-22 days, harvesting in time when the samara turns yellow-brown to mature from green.
6. Sowing and seedling raising: and after the fruits are harvested, removing impurities, and performing plug seeding, seedling raising and seeding within 2-3 days after the fruits are harvested. And (3) substrates of river sand, vegetable garden soil and special seedling peat are respectively adopted for hole-tray seeding and seedling raising, wherein one hole is formed in a 128-hole tray during seeding, water is poured after seeding, and the temperature of the substrate is kept at 20-25 ℃. And performing conventional seedling raising treatment in the later stage, keeping the substrate wet, and taking out of the soil for germination 4-7 days after sowing. The test results are shown in Table 4.
TABLE 4 Effect of different media on the germination and seedling development of Ulmus Davidiana Hance seeds
| Class of substrate | Time of sowing | Budding time (d) | The germination percentage is% | The seedling rate% |
| River sand | 4 month and 22 days | 4 month and 26 days | 91.2 | 80.3 |
| Common vegetable garden soil | 4 month and 22 days | 4 month and 29 days | 82.1 | 76.5 |
| Special peat for seedling culture | 4 month and 22 days | 4 month and 28 days | 98.6 | 95.3 |
7. Seedling stage management: (1) temperature management: the temperature is kept at about 25 ℃ in the day and preferably at 16-20 ℃ at night but not lower than 10 ℃ before the seedlings come out of the earth and cotyledons are completely unfolded, and the temperature can be adjusted by shading, ventilating, spraying water and the like. (2) And (3) water and fertilizer management: spraying small water with a spraying pot after seedling emergence, wetting the substrates in a dry mode, spraying foliar fertilizer for about 4 times by combining water spraying, and alternately spraying water-soluble fertilizers 20-10-20 and 14-0-14 (purchased from Hangzhou Hongyou flower Co., Ltd.) at a concentration of 50-100 mg/kg for 7 days. (3) Illumination: the illumination intensity is 1500-2500 Lux, and the illumination hours are 8-10 h/d; when the seedlings come out of the earth, gradually increasing the illumination intensity and the illumination hours of the seedbed by replacing a shading net or turning on a light supplement lamp, wherein the illumination intensity is increased from 1500Lux to 2500Lux, and the illumination hours are increased from 8h/d to 10 h/d; increasing 500Lux illumination intensity and 1h/d illumination hours every 7-10 days, wherein the maximum illumination intensity in a seedling stage is 2500Lux, and the maximum illumination hours is 10 h/d. (4) And (3) pest control: the prevention is mainly carried out, after the seedlings come out of the soil, 800 times of liquid carbendazim or thiophanate methyl is sprayed for 7-10 days on average alternately, and the seedbed is often ventilated.
8. Transplanting and potting: and (3) transplanting the seedlings into a pot in time when the seedlings grow 3-4 true leaves (the height reaches 2-6 cm).
Claims (5)
1. The in vitro cultivation method of the elm longituba fruits is characterized by mainly comprising the following steps:
1) selecting a mother tree: selecting a strong, straight and disease and insect pest-free mother tree, and planting the mother tree for 20-30 years;
2) branch collection: when the flower buds of elm occur but the inflorescence is not formed, high-branch scissors are used for shearing branches with the flower buds, the branches are taken back to a laboratory, and the flower buds on the branch sections are required to be full and healthy;
3) in vitro cultivation: cutting the collected branch sections into branch sections with the length of 30-40 cm, inserting the branch sections into a container bottle for liquid cultivation, and keeping the environmental temperature at 10-20 ℃ and the humidity at 70-80%; replacing the culture solution every 5-7 days; the liquid cultivation is nutrient solution cultivation, and the formula of the nutrient solution is as follows: potassium nitrate 607 mg/L, calcium nitrate 945 mg/L, ammonium phosphate 115 mg/L, magnesium sulfate 493 mg/L, GGR 6 No. 50 mg/L, aspirin 100 mg/L, activated carbon 2 mg/L, iron salt solution 2.5 ml/L, trace element solution 5ml/L, pH = 6.0; the iron salt solution comprises the following formula: 2.78 g of ferrous sulfate heptahydrate, 3.73 g of ethylenediamine disodium acetate and 500 ml of distilled water; the formula of the trace element solution is as follows: potassium iodide 0.83mg/L, boric acid 6.2 mg/L, manganese sulfate 22.3 mg/L, zinc sulfate 8.6 mg/L, sodium molybdate 0.25 mg/L, copper sulfate 0.025mg/L, cobalt chloride 0.025 mg/L;
4) harvesting fruits: when the samara turns yellow-brown to ripe from green, the samara is collected in time.
2. The sowing and seedling raising method for elms in the long-ordered market is characterized by comprising the following steps:
1) sowing: removing impurities from the ulmus davidiana fruits according to claim 1, performing plug seeding, seedling raising and seeding within 2-3 days after the fruits are harvested, watering thoroughly after seeding, and keeping the temperature of a matrix at 20-25 ℃;
2) seedling stage management: mainly comprises temperature management, water and fertilizer management, illumination control and pest control; the temperature management is as follows: keeping the temperature at about 25 ℃ in the day and preferably at 16-20 ℃ at night but not lower than 10 ℃ before the seedlings come out of the earth and cotyledons are completely unfolded, and adjusting the temperature by shading, ventilating and spraying water;
3) transplanting and potting: and (5) transplanting the seedlings into a pot in time when 3-4 true leaves grow out of the seedlings.
3. The elm planting and seedling method according to claim 2, wherein the water and fertilizer management method in step 2) is: spraying small water with a spraying pot after seedling emergence, wetting the substrates in a dry and wet mode, spraying foliar fertilizer for about 4 times by combining water spraying, and alternately spraying water-soluble fertilizers 20-10-20 and 14-0-14 at the concentration of 50-100 mg/kg for 7 days.
4. The elm sowing and seedling raising method according to claim 2, wherein the illumination control method in step 2) is as follows: after the seedlings come out of the soil, the illumination intensity and the illumination hours are gradually increased, wherein the illumination intensity is 1500-2500 Lux, and the illumination hours are 8-10 h/d.
5. The elm sowing and seedling raising method according to claim 2, wherein the pest control method in step 2) is: after the seedlings come out of the soil, 800 times of liquid carbendazim or thiophanate methyl is alternately sprayed once in 7-10 days on average, and the seedbed is often ventilated.
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