CN108496600B - Micro-cuttage rapid propagation method for virus-free rehmannia test-tube plantlets - Google Patents
Micro-cuttage rapid propagation method for virus-free rehmannia test-tube plantlets Download PDFInfo
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- CN108496600B CN108496600B CN201810375122.7A CN201810375122A CN108496600B CN 108496600 B CN108496600 B CN 108496600B CN 201810375122 A CN201810375122 A CN 201810375122A CN 108496600 B CN108496600 B CN 108496600B
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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
- A01G2/00—Vegetative propagation
- A01G2/10—Vegetative propagation by means of cuttings
Abstract
The invention particularly relates to a micro-cuttage rapid propagation method for virus-free rehmannia test-tube plantlets, which solves the problems of low operation efficiency, high cost and easy death of the existing rehmannia test-tube plantlet propagation. a. Cutting the detoxified rehmannia test-tube plantlet into stem segments with buds for culture at the culture temperature of 25-28 ℃, the illumination intensity of 2500Lx-3000Lx and the illumination time of 8 hours/day; b. cutting into stem sections with buds in an open environment, and immediately soaking in tap water; c. cutting the seedlings in a seedling tray, wherein two stem sections with buds are arranged in each hole, the cutting positions are distributed in a diagonal shape, the humidity in the seedling tray is 90-95 percent before new buds grow on the stem sections with the buds, the temperature is 15-30 ℃, and the illumination intensity under the film is 1500 Lx-2000 Lx; d. after the cutting seedling grows new buds and roots, transplanting the cutting seedling into a field to finish rapid propagation. The invention greatly improves the propagation quantity and reduces the cost of rapid propagation of the virus-free rehmannia test-tube plantlet.
Description
Technical Field
The invention relates to a test-tube plantlet propagation method in agricultural planting technology, in particular to a micro-cuttage rapid propagation method of a virus-free rehmannia test-tube plantlet.
Background
The Chinese medicinal material dosage of the rehmannia is large, the annual planting area of China is about 60 ten thousand mu, and according to investigation, the rehmannia planting area in the south-of-the-Shang region currently accounts for more than 50% of the rehmannia planting area in China. However, the rehmannia glutinosa is asexually propagated by root blocks in production for a long time, and is damaged by pathogenic bacteria such as rehmannia glutinosa degeneration virus (DDV), rehmannia glutinosa X virus and CMV, so that the rehmannia glutinosa is degenerated in nature, and the yield and the quality are seriously affected. For a long time, research institutes or universities continuously have produced detoxified seedlings by stem tip detoxification and test-tube seedling rapid propagation technology.
The technology obviously improves the yield and the quality of the rehmannia greatly, but the bottleneck problem which is difficult to remove is as follows: 1. the operation efficiency is low, and the operation speed is influenced by the requirement of aseptic operation; 2. the cost of raw materials is high, the requirements of test-tube plantlets on the components of a culture medium are high, and a large number of culture bottles are needed; 3. the house consumption is large, ineffective glass seedlings are easy to grow out by test tube culture, and in addition, the test tube seedlings die due to bacterial pollution; 4. a complicated transition process is required, and the maximum temperature at the initial transition cannot exceed 28 ℃, which results in a large number of deaths with slight carelessness.
Therefore, although the detoxified rehmannia has a plurality of advantages, the effect is not ideal after the related departments of all levels of governments in China and places put into practice for many years, the rehmannia seedlings used in large quantities in the production of the rehmannia still are self-bred and planted by farmers at present, and the application of the detoxified seedlings is very little.
Disclosure of Invention
The invention provides a micro-cuttage rapid propagation method for a virus-free rehmannia test-tube plantlet, aiming at solving the problems of low operation efficiency, high cost and easy death of the existing rehmannia test-tube plantlet propagation.
The invention is realized by adopting the following technical scheme: a micro-cuttage rapid propagation method of a detoxified rehmannia test-tube plantlet comprises the following steps: a. cutting the detoxified rehmannia test-tube plantlet into 2-2.5cm stem segments with buds on an ultraclean workbench, and then transferring the stem segments into a culture medium containing 30g/L, PH of sugar with the value of 5.6-6.4 for culture at the temperature of 25-28 ℃, the illumination intensity of 2500Lx-3000Lx and the illumination time of 8 hours/day; b. when the detoxicated rehmannia root test-tube plantlet is cultured for 30-35 days or grows to more than 10cm, cutting into a stem section with buds, which has one leaf and one bud, by using scissors at an angle of 45 degrees in an open environment, and immediately soaking in tap water at 10-25 ℃ for more than or equal to 2 min; c. the soaked stem sections with buds are inserted into a seedling tray filled with vermiculite, two stem sections with buds are arranged in each hole, the inserting positions are distributed in a diagonal shape, the humidity in the seedling tray is 90-95 percent before new buds grow out of the stem sections with buds, the temperature is 15-30 ℃, and the illumination intensity under the film is 1500 Lx-2000 Lx; d. after the cutting seedling grows new buds and roots, transplanting the cutting seedling into a field to finish rapid propagation.
According to the invention, the test-tube plantlets are cut again after being cut and cultured, and then are bred in a cutting seedling mode, so that the breeding amount is greatly increased, and the breeding mode of combining test tubes and non-test tubes is adopted, so that the obtained virus-free plantlets are more robust, can be directly transplanted without transition, and meanwhile, the problem that the conventional rehmannia test-tube plantlet breeding is low in operation efficiency, high in cost and easy to die is solved.
The humidity control in the seedling tray adopts the following method: and (3) building a simple arched shed, covering a single-layer film, pricking a plurality of small holes on the film when the humidity is more than 95%, and slowly watering from the edge of the arched shed when the humidity is less than 90%.
The humidity control method is more convenient and faster to operate.
The invention greatly improves the propagation quantity, compared with the traditional propagation quantity of whole plant transplanting of the test-tube plantlet, the propagation quantity is more than 25 times, the cost is only below the traditional 1/5, the obtained virus-free plantlet is more robust, can be directly transplanted to a field without excess, and can normally propagate the micro-cutting plantlet at the outdoor temperature of more than 28 ℃, thereby greatly prolonging the propagation time of the virus-free plantlet, further avoiding the occurrence of the pollution of the test-tube plantlet by bacteria or vitrification phenomenon, operating the micro-cutting part in an open environment without sterile conditions, being simple and easy to implement, and having the advantages of simple operation, low cost, small management difficulty and suitability for industrialized propagation.
Table 1 shows the factors and levels (mg/L) of the rehmannia glutinosa test-tube plantlet culture survival rate test, and an orthogonal design test is adopted to select the optimal combination of the three factors. 10 bottles (4/bottle) of detoxified rehmannia test-tube plantlets were selected for each test number.
TABLE 1
Table 2 shows the combination of the factors and the results of the rehmannia glutinosa test-tube plantlet culture survival rate test, wherein A in Table 2 indicates the temperature, B indicates the illumination, and C indicates the illumination time.
As can be seen from Table 2, when the maximum survival rate was 0.96, the culture temperature was 25 ℃ to 28 ℃, the illumination was 2500-.
Table 3 shows the survival rate test factors and levels (mg/L) of rehmannia glutinosa libosch micro-cutting seedlings, and three important factors influencing the rehmannia glutinosa libosch cutting seedlings are selected by adopting orthogonal design to obtain the optimal combination of the three factors. And selecting micro-cutting seedlings of 2 seedling raising plates as objects for each test number.
When the density of the cutting seedlings is 800-; when 1400-2000 plants/square meter are used, every hole in the seedling raising plate is two, and the seedlings are cut by diagonal lines; 2000-2600 plants/square meter, three in each hole of the seedling-raising tray, and cutting in equilateral triangle.
Table 4 shows the combination of the factors and the results of the rehmannia root micro-cutting seedling survival rate test, in table 4, a indicates humidity, B indicates under-film illumination, and C indicates cutting density.
As can be seen from Table 4, when the highest survival rate is 0.98, the humidity of the seedlings is 90% -95%, the illumination under the film is 1500-.
Table 5 is the cost accounting.
Detailed Description
Example 1
A micro-cuttage rapid propagation method of a detoxified rehmannia test-tube plantlet comprises the following steps: a. cutting the detoxified rehmannia test-tube plantlet into 2cm stem segments with buds on a superclean workbench, and then transferring the cut detoxified rehmannia test-tube plantlet into a culture medium containing 30g/L, PH of sugar with the value of 5.6 for culture at the culture temperature of 25 ℃, the illumination intensity of 2500Lx and the illumination time of 8 hours/day; b. when the detoxified rehmannia test-tube plantlet is cultured for 30 days or grows to more than 10cm, cutting into a stem section with buds with one leaf and one bud by using scissors at an angle of 45 degrees in an open environment, and immediately soaking in tap water at 10 ℃ for 2 min; c. cutting the soaked stem segments with buds into a seedling tray filled with vermiculite, wherein two stem segments with buds are arranged in each hole, the cutting positions are distributed in a diagonal manner, the humidity in the seedling tray is 90 percent before new buds grow on the stem segments with buds, the temperature is 15 ℃, and the illumination intensity under the film is 1500 Lx; d. after the cutting seedling grows new buds and roots, transplanting the cutting seedling into a field to finish rapid propagation.
The humidity control in the seedling tray adopts the following method: and (3) building a simple arched shed, covering a single-layer film, pricking a plurality of small holes on the film when the humidity is more than 95%, and slowly watering from the edge of the arched shed when the humidity is less than 90%.
Example 2
A micro-cuttage rapid propagation method of a detoxified rehmannia test-tube plantlet comprises the following steps: a. cutting the detoxified rehmannia test-tube plantlet into 2.5cm stem segments with buds on an ultraclean workbench, and then transferring the stem segments into a culture medium containing 30g/L, PH of sugar with the value of 6.4 for culture, wherein the culture temperature is 28 ℃, the illumination intensity is 3000Lx, and the illumination time is 8 hours/day; b. when the detoxified rehmannia test-tube plantlet is cultured for 35 days or grows to more than 10cm, cutting into a stem section with buds with one leaf and one bud by using scissors at an angle of 45 degrees in an open environment, and immediately soaking in tap water at 25 ℃ for 8 min; c. cutting the soaked stem segments with buds into a seedling tray filled with vermiculite, wherein two stem segments with buds are arranged in each hole, the cutting positions are distributed in a diagonal manner, the humidity in the seedling tray is 95 percent, the temperature is 30 ℃, and the illumination intensity under the film is 2000Lx before new buds grow out of the stem segments with buds; d. after the cutting seedling grows new buds and roots, transplanting the cutting seedling into a field to finish rapid propagation.
The humidity control in the seedling tray adopts the following method: and (3) building a simple arched shed, covering a single-layer film, pricking a plurality of small holes on the film when the humidity is more than 95%, and slowly watering from the edge of the arched shed when the humidity is less than 90%.
Example 3
A micro-cuttage rapid propagation method of a detoxified rehmannia test-tube plantlet comprises the following steps: a. cutting the detoxified rehmannia test-tube plantlet into 2.1cm stem segments with buds on an ultraclean workbench, and then transferring the stem segments into a culture medium containing 30g/L, PH of sugar with the value of 5.9 for culture, wherein the culture temperature is 27 ℃, the illumination intensity is 2699Lx, and the illumination time is 8 hours/day; b. when the detoxified rehmannia test-tube plantlet is cultured for 32 days or grows to more than 10cm, cutting into a stem section with buds with one leaf and one bud by using scissors at an angle of 45 degrees in an open environment, and immediately soaking in tap water at 19 ℃ for 5 min; c. cutting the soaked stem segments with buds into a seedling tray filled with vermiculite, wherein two stem segments with buds are arranged in each hole, the cutting positions are distributed in a diagonal manner, the humidity in the seedling tray is 92 percent, the temperature is 22 ℃, and the illumination intensity under the film is 1846Lx before new buds grow out of the stem segments with buds; d. after the cutting seedling grows new buds and roots, transplanting the cutting seedling into a field to finish rapid propagation.
The humidity control in the seedling tray adopts the following method: and (3) building a simple arched shed, covering a single-layer film, pricking a plurality of small holes on the film when the humidity is more than 95%, and slowly watering from the edge of the arched shed when the humidity is less than 90%.
In the specific implementation process, the size of the seedling raising tray is 540 multiplied by 280 multiplied by 55mm and 32 holes, the size of the simple arched shed is 2m wide multiplied by 10m long multiplied by 1m high, the interval of each bamboo pole or plastic pipe is 1.5m, and the standard of seedling formation is that a stem section with buds grows 1-2 leaves and roots with the length of more than 1 cm.
Claims (2)
1. A micro-cuttage rapid propagation method of detoxified rehmannia test-tube plantlets is characterized in that: the method comprises the following steps:
a. cutting the detoxified rehmannia test-tube plantlet into 2-2.5cm stem segments with buds on an ultraclean workbench, and then transferring the stem segments into a culture medium containing 30g/L, PH of sugar with the value of 5.6-6.4 for culture at the temperature of 25-28 ℃, the illumination intensity of 2500Lx-3000Lx and the illumination time of 8 hours/day; b. when the detoxicated rehmannia root test-tube plantlet is cultured for 30-35 days or grows to more than 10cm, cutting into a stem section with buds, which has one leaf and one bud, by using scissors at an angle of 45 degrees in an open environment, and immediately soaking in tap water at 10-25 ℃ for more than or equal to 2 min; c. the soaked stem sections with buds are inserted into a seedling tray filled with vermiculite, two stem sections with buds are arranged in each hole, the inserting positions are distributed in a diagonal shape, the humidity in the seedling tray is 90-95 percent before new buds grow out of the stem sections with buds, the temperature is 15-30 ℃, and the illumination intensity under the film is 1500 Lx-2000 Lx; d. after the cutting seedling grows new buds and roots, transplanting the cutting seedling into a field to finish rapid propagation.
2. The micro-cutting rapid propagation method of the detoxified rehmannia test-tube plantlet according to claim 1, characterized in that: the humidity control in the seedling tray adopts the following method: and (3) building a simple arched shed, covering a single-layer film, pricking a plurality of small holes on the film when the humidity is more than 95%, and slowly watering from the edge of the arched shed when the humidity is less than 90%.
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Inventor after: Zuo Jingjing Inventor after: Yan Guiyun Inventor after: Huo Liguang Inventor after: Wang Huijie Inventor after: Liu Shaoxiang Inventor after: Yan Jianjun Inventor after: Zuo Min Inventor before: Zuo Jingjing |
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