CN114946662A - Method for rapid propagation by using stems of thinleaf cypress branches - Google Patents
Method for rapid propagation by using stems of thinleaf cypress branches Download PDFInfo
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- CN114946662A CN114946662A CN202210757345.6A CN202210757345A CN114946662A CN 114946662 A CN114946662 A CN 114946662A CN 202210757345 A CN202210757345 A CN 202210757345A CN 114946662 A CN114946662 A CN 114946662A
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- 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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Abstract
The invention provides a method for rapid propagation by using stems of cypress with thinner flowers, which comprises the following steps: s1, selecting annual stems of the cypress branches as explants, and disinfecting; s2, placing the explant in GA 3 Soaking in the solution, wrapping with preservative film, and sequentially performing low-temperature treatment and high-temperature treatment; s3, preparing a culture substrate, and paving and covering the explant on the substrate; and S4, maintaining until sprout seedlings grow out, digging out the buried explants and cutting the buried explants after the sprout seedlings are semi-lignified, and finally transplanting the cut explants in the field. The invention can shorten the dormancy time of the explant after disinfection and induction treatment, can reach the transplanting standard in about 18d, greatly shortens the breeding and seedling period, takes the stem as the explant, is not limited by time and seasons, can breed at any time, and can provide a large amount of plant materials for the treatment of hydro-fluctuation belts in three gorges reservoir areas.
Description
Technical Field
The invention belongs to the technical field of propagation of threatened plants, and particularly relates to a method for rapid propagation by using stems of thinly-flowered cypress branches.
Background
The flowering cypress branch (Myricarialaxlora) is a perennial shrub of the genus cypress of the family Tamaricaceae, and the three gorges reservoir area is the core distribution area. The special plants in the three gorges reservoir area which are possibly affected and endangered to be extinct are mentioned in the environmental impact report of the Yangtze river three gorges hydro junction, and meanwhile, the special plants are national secondary important protection wild plants, and have great development and application values in the aspect of treatment of hydro-fluctuation belts.
At present, researches on the cypress branch of the hydrophobic flower mainly focus on community structure, morphological structure, seed germination, artificial breeding, photosynthetic physiological and biochemical characteristics and the like. At present, research on cuttage, sowing and breeding of the cypress branches of the cypress has been carried out by existing scientific research institutions, but the problems of easy loss of viability of the cypress branch seeds at normal temperature, seasonal limitation of growth and breeding and the like have high requirements on breeding technical conditions and time, and the problems of low utilization efficiency of branches and stems and small breeding coefficient of cuttage breeding exist. The existing breeding technology restricts the rapid propagation and protection of the cypress branch of the flowering Chinese arborvitae to a certain extent.
Disclosure of Invention
The invention provides a method for rapid propagation by utilizing the stems of the cypress branches of the thinning flower, which has the advantages of low technical threshold, high emergence rate, short propagation time, no season limitation and convenience for large-scale popularization.
The technical scheme of the invention is that a method for rapid propagation by using the stems of the thinleaf cypress branches comprises the following steps:
s1, selecting annual stems of the cypress branches as explants, and disinfecting;
s2, placing the explant in GA 3 Soaking in the solution, wrapping with preservative film, and sequentially performing low-temperature treatment and high-temperature treatment;
s3, preparing a culture medium, adding a potassium permanganate solution, uniformly mixing, disinfecting and replenishing water, paving stone grains, peat soil and river sand in sequence from bottom to top, paving the explant on the river sand, covering the river sand, and finally covering pine flakes;
s4, preserving heat and moisture, maintaining until sprouts grow out, digging out the buried explants and cutting the buried explants after the sprouts are semi-lignified to ensure that each stem segment has a sprout, and finally transplanting in the field.
Further, selecting strong and disease and insect pest-free flower-thinning cypress branches as mother plants in S1, collecting plump stems on the 1-year-old land,
further, before the sterilization treatment in S1, the leaves on the stem are removed, and the stem is cut into stem segments with the length of 20-30 cm to be used as explants.
Further, in the sterilization treatment in S1, the mixture is soaked in a potassium permanganate solution with the mass concentration of 0.2% for 7-10min for sterilization, and then is washed with sterile water for 10-15 min.
Further, the concentration of the GA3 solution in the S2 is 150-250mg/L, and the soaking time is 2-3 h.
Further, the temperature of the low-temperature treatment in S2 is 4-6 ℃, and the time is 12-15 h; the high temperature treatment is carried out at 30-35 deg.C for 12-15 hr.
Further, 0.2-0.3% of potassium permanganate solution with mass concentration is added into the culture medium in S3 and is mixed evenly, and the water content of river sand is 60% -70%.
Further, the thickness of the stone particles in the S3 is 2-4cm, peat soil with the thickness of 1-2cm is paved, and river sand with the thickness of 3-4cm is paved; putting the explant and then covering river sand with the thickness of 3-3.5 cm.
Further, when the explants are tiled in S3, they are laid parallel to each other and spaced 2-4cm apart.
Further, during the culture in S4, covering an arched plastic film, keeping the temperature at 20-25 ℃, spraying sterile water to keep the substrate moist, and taking the standard that the substrate is held by hand to form a mass and is loose but not scattered.
The invention has the following beneficial effects:
1. compared with the cutting propagation of the flowering cypress branches, the invention takes the stems as the explants, but the invention fully processes the explants in the previous period. Soaking in 0.2% potassium permanganate solution for 7-10min to kill pathogenic bacteria on the surface of stem, and activating enzyme activity in seed body with manganese as trace elementThe seedlings grow regularly and robustly; it is placed at GA 3 Soaking in the solution to increase the content of auxin in the stems and promote the rapid growth of the stems and leaves; and the stem is wrapped by a preservative film and then sequentially subjected to low-temperature and high-temperature variable-temperature activation treatment, so that the content of inhibiting substances in the stem is reduced, dormancy is broken, and the stem germination is promoted.
2. The invention can shorten the dormancy time of the explant after disinfection and induction treatment, can reach the transplanting standard in about 18d, greatly shortens the breeding and seedling period, takes the stem as the explant, is not limited by time and seasons, can breed at any time, and can provide a large amount of plant materials for the treatment of hydro-fluctuation belts in three gorges reservoir areas. .
3. The rapid propagation method provided by the invention has high propagation coefficient, one stem segment can germinate a plurality of bud seedlings after long stem segment treatment, the propagation speed is high, the seedlings emerge neatly, the root system is flourishing, and the survival rate of the seedlings can reach more than 90% after the seedlings are transplanted by plants.
4. Stone grains with the thickness of 2-4cm are laid on the bottom layer of the culture substrate to facilitate drainage, so that the substrate can be prevented from being anoxic due to excessive water accumulation, the respiration of stems is hindered, and the physiological function is reduced; the peat soil with the thickness of 1-2cm is laid on the stone particles, so that the temperature of the matrix can be maintained in a higher and stable range, and sufficient nutrients can be provided for the growth of the root system after the stems take root; the river sand paved with the stems up and down has the effects of ventilation and drainage, gaps of the river sand are moderate in size, and the advantages of ventilation and drainage are achieved.
5. The stems are directly buried under river sand, if the stems are dormant for a long time, the stems can miss the window period of sprouting and gradually lose vitality, and finally become rotten and mildewed. The invention is sterilized by potassium permanganate and GA 3 Soaking in solution, activating at high and low temperature, burying the stem in layered matrix, and exciting adventitious bud and adventitious root. Compared with the limited fixed buds growing from the stem tip or leaf axilla and other positions in the cuttage, the propagation number of the buds per unit length is increased, the propagation coefficient is increased, and the propagation number reachesTo the aim of rapid propagation.
Drawings
FIG. 1 is a photograph after 10 days of treatment in example 1.
FIG. 2 is a photograph after 18 days of treatment in example 1.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention.
Example 1: method for rapid propagation by using stems of thinleaf cypress branches
1) Selecting strong and disease and pest free flower thinning cypress branches as mother plants, and collecting plump 1-year-old stems of the cypress branches. And then, preprocessing the stems, removing leaves on the stems, and cutting the stems into stem sections with the length of 20-30 cm. And (3) soaking the cut stem sections in a potassium permanganate solution with the mass concentration of 0.2% for 8min for disinfection, fishing out after the disinfection, and washing with sterile water for 10 min.
2) After the pretreatment is finished, the stems are initiated. The stem segments were placed in a 200mg/L GA concentration 3 Soaking in the solution for 2 h. Taking out, wrapping the stem sections with a preservative film, and placing in a constant-temperature incubator at 5 ℃ for low-temperature treatment for 12 h; after the treatment, the temperature is adjusted to 30 ℃ for high-temperature treatment for 12 h.
3) And (3) finishing the initiation bed, spraying a potassium permanganate solution with the mass concentration of 0.2% into the clean coarse stone particles, the clean fine river sand and the clean peat soil, and uniformly stirring and fully disinfecting. The water content of the fine river sand is ensured to be 60 percent, and the standard of 'conglobation by hand holding and loosening without scattering' is achieved. Coarse stone grains with the thickness of 2cm are paved at the bottom of the bed, peat soil with the thickness of 1cm is paved, and fine river sand with the thickness of 3cm is paved.
4) And spreading the cut stem sections on the fine river sand, wherein the stem sections are parallel to each other and are spaced by 2 cm. Covering fine river sand of 3cm on the laid stem section, and laying a layer of loose scale on the fine river sand. Covering a layer of arched plastic film on the initiation bed, maintaining the temperature, checking every 3d, spraying appropriate amount of sterile water to keep the matrix moist, and controlling the temperature of the initiation bed at 20-25 deg.C. After 10d and 18d of treatment, the stem section photos are shown in figures 1 and 2, after the bud seedlings are semi-lignified, the buried stem sections are dug out and cut, and each stem section is guaranteed to have one bud seedling, so that field transplanting can be carried out; and 7 sprouts germinate on average in each stem section.
Example 2:
referring to example 1, other processing conditions were not changed, and GA was used 3 The solution was set at two concentrations of 150mg/L and 250mg/L, checked every 3d for 1 month, and the results were recorded as in Table 1 below.
TABLE 1
| Numbering | GA 3 Concentration of solution | Time of germination | Average number of sprouts per stem segment | Root growth conditions |
| Example 2-1 | 150mg/L | 12d | 7 are provided with | Vigorous growth of root system |
| Examples 2 to 2 | 250mg/L | 9d | 6 are | Vigorous growth of root system |
Example 3:
referring to example 1, the following low temperature and high temperature treatments were set without changing other treatment conditions, and the following treatments were examined every 3d for 1 month, and the experimental effects were recorded as shown in table 2 below.
TABLE 2
| Numbering | Temperature-changing treatment | Time of germination | Average number of sprouts per stem segment | Root growth conditions |
| Example 3-1 | Low temperature 4 deg.C/high temperature 32 deg.C | 9d | 7 are provided with | Vigorous growth of root system |
| Examples 3 to 2 | Low temperature 4 deg.C/high temperature 35 deg.C | 12d | 6 are | Vigorous growth of root system |
| Examples 3 to 3 | Low temperature 6 deg.C/high temperature 32 deg.C | 12d | 6 are | Vigorous growth of root system |
| Examples 3 to 4 | Low temperature 6 deg.C/high temperature 35 deg.C | 12d | 7 are provided with | Vigorous growth of root system |
Example 4:
referring to example 1, the river sand coverage thickness was set to 2.5cm and 3.5cm for two treatments with the other treatment conditions unchanged, and the treatments were checked every 3d for 1 month, and the experimental effects were recorded as shown in table 3 below.
TABLE 3
| Numbering | Thickness of river sand coverage | Time of germination | Average number of sprouts per stem segment | Root growth conditions |
| Example 4-1 | 2.5cm | 9d | 6 are | Normal flourishing root system |
| Example 4 to 2 | 3.5cm | 12d | 7 are provided with | Vigorous growth of root system |
Comparative example 1:
referring to example 1, other processing conditions were not changed, and GA was used 3 The solution is set to be 100mg/L and 300mg/L, a blank control is additionally set to be soaked in clear water, the blank control is checked every 3d and continuously observed for 1 month, and the recorded experiment effects are shown in the following table 4.
TABLE 4
Comparative example 2:
referring to example 1, other treatment conditions were unchanged, and the substrate was subjected to two treatments, one is to mix river sand and peat uniformly, and the other is to use field conventional soil, and the soil was examined every 3d for 1 month, and the experimental effects were recorded as follows:
comparative example 3:
referring to example 1, the following low-temperature and high-temperature treatments were set without changing other treatment conditions, and were examined every 3d for 1 month, and the experimental effects were recorded as follows:
comparative example 4:
referring to the embodiment 1, the river sand coverage thickness is set to be 2cm and 4cm for two treatments with other treatment conditions unchanged, the treatments are checked every 3d for 1 month, and the recorded experimental effects are as follows:
| numbering | Thickness of river sand coverage | Time of germination | Average number of sprouts per stem segment | Root growth conditions |
| Comparative example 4-1 | 2cm | 21d | 2 are provided with | Normal growth of root system |
| Comparative examples 4 to 2 | 4cm | 27d | 1 is provided with | Weak root growth |
The above-described embodiments are merely preferred embodiments of the present invention, and should not be construed as limiting the present invention, and features in the embodiments and examples in the present application may be arbitrarily combined with each other without conflict. The protection scope of the present invention is defined by the claims, and includes equivalents of technical features of the claims. I.e., equivalent alterations and modifications within the scope hereof, are also intended to be within the scope of the invention.
Claims (10)
1. A method for rapid propagation by using the stems of the thinleaf cypress branches is characterized by comprising the following steps:
s1, selecting annual stems of the cypress branches as explants, and disinfecting;
s2, placing the explant in GA 3 Soaking in the solution, wrapping with preservative film, and sequentially performing low-temperature treatment and high-temperature treatment;
s3, preparing a culture medium, adding a potassium permanganate solution, uniformly mixing, disinfecting and replenishing water, paving stone grains, peat soil and river sand in sequence from bottom to top, paving the explant on the river sand, covering the river sand, and finally covering pine flakes;
s4, preserving heat and moisture until sprouts grow out, digging out the buried explants and cutting the buried explants after the sprouts are semi-lignified to ensure that each stem section has one sprout, and finally transplanting the sprouts in the field.
2. The method of claim 1, wherein: s1, selecting strong and disease and pest free white cypress branches as mother plants, and collecting plump 1-year-old stems of the white cypress branches.
3. The method of claim 1, wherein: and S1, before disinfection treatment, removing leaves on the stem, and cutting the stem into stem sections with the length of 20-30 cm to be used as explants.
4. The method of claim 1, wherein: and S1, soaking in 0.2% potassium permanganate solution for 7-10min for sterilization, and washing with sterile water for 10-15 min.
5. The method of claim 1, wherein: GA in S2 3 The concentration of the solution is 150-250mg/L, and the soaking time is 2-3 h.
6. The method of claim 5, wherein: the low-temperature treatment in S2 is carried out at 4-6 ℃ for 12-15 h; the high temperature treatment is carried out at 30-35 deg.C for 12-15 hr.
7. The method of claim 1, wherein: and adding 0.2-0.3% by mass of potassium permanganate solution into the culture medium in S3, and uniformly mixing until the water content of river sand is 60% -70%.
8. The method of claim 1, wherein: s3, paving peat soil with the thickness of 1-2cm and river sand with the thickness of 3-4cm, wherein the thickness of the stone grains is 2-4 cm; putting the explant and then covering river sand with the thickness of 3-3.5 cm.
9. The method of claim 8, wherein: when the explants in S3 are spread flatly, the explants are spread in parallel with a spacing of 2-4 cm.
10. The method of claim 1, wherein: when culturing in S4, covering with arched plastic film, keeping the temperature at 20-25 deg.C, spraying sterile water to keep the matrix moist, and making into mass with hand.
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Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| US20200093084A1 (en) * | 2012-12-20 | 2020-03-26 | MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. | Stable transformation of a population and a method of biocontainment using haploinsufficiency and underdominance principles |
| CN106472134A (en) * | 2016-09-18 | 2017-03-08 | 海南大学 | The method that a kind of falling zone muskeg recovers suitable species screening |
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