CN108464207B - Culture method for improving transplanting survival rate of rice tissue culture seedlings - Google Patents
Culture method for improving transplanting survival rate of rice tissue culture seedlings Download PDFInfo
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- CN108464207B CN108464207B CN201810331778.9A CN201810331778A CN108464207B CN 108464207 B CN108464207 B CN 108464207B CN 201810331778 A CN201810331778 A CN 201810331778A CN 108464207 B CN108464207 B CN 108464207B
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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
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/20—Cereals
- A01G22/22—Rice
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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
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
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Abstract
The invention discloses a culture method for improving the transplanting survival rate of rice tissue culture seedlings, which is completed by the steps of tissue culture seedling cleaning, hormone treatment, soil preparation, transplanting, environment management, water and fertilizer management and the like. The invention effectively solves the problem that the survival rate of the transplanted rice tissue culture seedlings is not guaranteed basically by combining the innovation of agricultural measures and the application technology of the plant growth hormone, the survival rate of the transplanted rice tissue culture seedlings is more than 85 percent after the technology is applied, the root systems of the transplanted rice tissue culture seedlings are developed, the plants are robust, and the earring rate and the maturing rate are obviously improved.
Description
Technical Field
The invention belongs to the technical field of rapid propagation of plants, and particularly relates to a culture method for improving the transplanting survival rate of rice tissue culture seedlings.
Background
Tissue culture is a technique in which an isolated organ (e.g., root, stem, leaf, stem segment, protoplast), tissue, or cell of a plant is placed in a culture medium under artificially created sterile conditions and placed in an appropriate environment for continuous culture to obtain cells, tissue, or individuals. The technology is widely applied to agriculture and biological and medical research. The rice tissue culture seedling is a rice genetic transformation seedling obtained by utilizing a tissue culture technology, generally grows in a sterilized and closed test tube or culture bottle, and grows under the conditions of strict light temperature and the like.
The tissue culture seedling of the rice is very young and tender, and is not easy to survive after being transplanted (moved out of a closed test tube or a culture bottle). By applying the traditional method, the survival rate of the transplanted tissue culture seedlings of the rice is basically not guaranteed, and the survival rate of the transplanted tissue culture seedlings is less than 50% under the general condition. Even if the rice is survived, the growth of the rice at the later stage is sluggish, the ear forming rate is low, and the ears are small.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to solve the technical problem that the transplanted tissue culture seedlings of rice are not easy to survive, and provides the culture method for improving the transplanting survival rate of the tissue culture seedlings of rice.
The invention is realized by the following technical scheme:
a culture method for improving the transplanting survival rate of rice tissue culture seedlings comprises the following steps:
1) cleaning of tissue culture seedlings
Soaking the tissue culture seedlings for 2 days by using a nutrient solution formula of international rice, and then cleaning the tissue culture seedlings by using natural water at the temperature of 20-25 ℃, so as to remove the nutrient solution at the roots of the tissue culture seedlings and prevent the root systems of the tissue culture seedlings from mildewing after transplanting;
2) hormone treatment
Placing the cleaned tissue culture seedling into a test tube, adding natural water containing abscisic acid to submerge the root, culturing for 1-2 days under the condition of shade in a greenhouse, spraying gibberellin solution on the leaf surface of the seedling before transplanting, keeping the temperature and shading by using a black gauze;
3) soil preparation
Filling 7-10 kg of dry soil in each barrel of paddy field soil, adding water after filling the soil, mashing the soil uniformly, and standing the soil in a greenhouse for 2 days so as to balance soil heating and floating soil precipitation;
4) transplantation and environmental management
Transplanting, namely transplanting the seeds singly, wherein the transplanting depth is proper for the soil to just cover the roots of the seedlings, irrigating 1-2 cm of shallow water after transplanting the seedlings, shading the seedlings for one week by using a black gauze, and then removing the black gauze to culture the seedlings under the natural illumination of a greenhouse;
5) liquid manure management
And (4) irrigating shallow water and applying 10-15 grains/pot of special compound fertilizer for rice 4 weeks after transplanting, naturally drying, keeping a dry-wet alternate soil environment, and applying fertilizer for one week to plant the rice in the field.
The natural water is tap water which is exposed to the sun for 5 days.
The natural water contains 10-100 mg/L of abscisic acid.
The concentration of the gibberellin solution is 1-10 mg/L, and the spraying condition is that 2-5 ml of gibberellin solution is sprayed on the leaf surface of each seedling.
The shading condition of the black gauze is 1/10 light intensity reaching normal light so as to avoid direct sunlight.
The transplanting and environmental management also comprises the steps of adding shallow water to cultivate soil once after 15 days of transplanting, naturally drying the cultivated soil until the soil has tiny cracks, adding water, and then maintaining a dry-wet alternative soil environment.
The invention has the beneficial effects that:
the technology effectively solves the problems by combining the innovation of agricultural measures and the application technology of the plant growth hormone, the survival rate of the transplanted rice tissue culture seedlings is over 85 percent after the technology is applied, the root systems of the transplanted rice tissue culture seedlings are developed, the plants are robust, and the earring rate and the maturing rate are obviously improved.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to specific embodiments of the present invention, and it should be understood 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.
Example 1
The embodiment provides a culture method for improving the transplanting survival rate of the tissue culture seedlings of the rice aiming at the technical defects that the survival rate of the tissue culture seedlings of the rice is not guaranteed basically, the survival rate after the transplanting is less than 50% under the general condition, and the late growth is slow, the earring rate is low and the earring is small even if the tissue culture seedlings survive, and the culture method effectively solves the problems and is completed through the following steps:
1) cleaning of tissue culture seedlings
Soaking the tissue culture seedling with nutrient solution for 2 days (formula of nutrient solution of International Rice institute), cleaning with 20-25 deg.C natural water (tap water sun-dried for 5 days), removing culture medium (agarose, etc.) at the root of the tissue culture seedling, preventing the root system of the tissue culture seedling from mildewing after transplanting, and adding nutrient solution to submerge the root.
2) Hormone treatment:
placing the cleaned tissue culture seedlings (step 1) into a test tube, adding natural water to submerge roots, wherein the added natural water contains 10-100 mg/L abscisic acid (ABA), and adapting to greenhouse shading conditions for 1-2 days before transplanting. 5 hours before transplanting, 2-5 ml of gibberellin solution of 1-10 mg/L is sprayed on the leaf surface of each seedling, the environmental temperature is maintained at 20-25 ℃, and the seedlings are shaded by a black gauze (the illumination intensity of 1/10 of normal illumination is achieved, so that direct sunlight is avoided).
3) Soil preparation
The depth of the pot barrel is 30-50 cm, and the diameter is 30-40 cm. And (3) filling 7-10 kg of dry soil (the dry soil is 10-15 cm away from the top end of the water bucket) in each bucket of paddy field soil, adding water after filling the soil, mashing the soil uniformly, standing the soil in a greenhouse for 2 days so as to balance soil heating and floating soil precipitation, and keeping the temperature of the greenhouse at 20-25 ℃.
4) Transplantation and environmental management
Transplanting test-tube seedlings, transplanting the test-tube seedlings, wherein the transplanting depth is proper for soil to just cover the roots of the seedlings, irrigating shallow water of 1-2 cm after transplanting the seedlings, shading the seedlings with a black gauze (reaching the illumination intensity of 1/10 in normal illumination to avoid direct sunlight) for one week, and then uncovering the black gauze to culture the seedlings under the natural illumination (20-25 ℃) of a greenhouse. After 15 days of transplantation, shallow water is added for weeding (loosening the soil) once to ventilate and enrich oxygen, simultaneously, the rotten branches and leaves on the rice plants are removed, after weeding, the soil naturally falls to be dry until the soil has tiny cracks, then water is added, and then the dry-wet alternate soil environment is maintained. If the natural temperature of the atmosphere is in the range of 20-30, the potted plant can be moved to the outdoor natural environment for growing.
5) Liquid manure management
After 4 weeks after transplanting, the tissue culture seedlings are adapted to a new environment and begin to normally grow and tillere, shallow water is irrigated, 10-15 grains/pot of special compound fertilizer for paddy rice are applied, and the soil environment with alternate dry and wet is maintained after natural drying. After one week of fertilization, the seedlings can be transplanted to a field for planting (suitable for the condition of more seedlings), and can also be kept potted for planting (suitable for the condition of less seedlings), and the management of water, fertilizer, plant diseases, insect pests and weeds is similar to the production of a common field until the seedlings are harvested.
Example 2
In 2017, 7 batches of tissue culture seedlings are transplanted in a Fuyang base of the China Rice research institute, the transplanted seedlings are transplanted in a temperature-controlled greenhouse, the temperature control range of the greenhouse is 20-30 ℃, and the greenhouse is naturally illuminated and provided with a shading net. Soaking roots for 2 days by using 30mg/L abscisic acid (ABA) solution, and spraying 2ml of 3mg/L gibberellin solution on the leaf surfaces of each seedling 5 hours before transplanting. The results are shown in tables 1 and 2.
TABLE 1 survival rate of tissue culture seedlings after 20 days of transplantation
| Numbering batches | Number of tissue culture seedlings | Number of survival seedlings after transplantation | Survival rate after 20 days of transplantation |
| CN-170215-Pr-67 | 55 | 48 | 87.3 |
| CN-170325-Pr-59 | 80 | 68 | 85.0 |
| CN-170412-Pr-59 | 60 | 55 | 91.7 |
| CN-170412-Ar-67 | 91 | 80 | 87.9 |
| CN-170509-Si-59 | 95 | 83 | 87.4 |
| CN-170509-Ir-59 | 45 | 39 | 86.7 |
| CN-170606-Sn-59 | 174 | 161 | 92.5 |
| Total of | 600 | 534 | The average survival rate is 89.0 percent |
As can be seen from Table 1, the survival rate of the tissue culture seedlings 20 days after the transplantation by applying the technology is 85-92.5%, and the average survival rate is 89.0%.
TABLE 2 panicle formation and seed set (per cluster) for the genetic transformation lines of Nipponbare rice
As can be seen from Table 2, the heading rate of the tissue culture seedlings transplanted by the conventional technology is 41.5-48.4, and the average heading rate is 44.5%; the heading rate of the tissue culture seedlings transplanted by applying the technology is 60.0-68.1%, and the average heading rate is 62.7%. The setting percentage is increased from 25.8-39.7% to 60.7-80.4% and the average setting percentage is also increased from 29.9% to 67.2%.
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 (4)
1. A culture method for improving the transplanting survival rate of rice tissue culture seedlings is characterized by comprising the following steps:
1) cleaning of tissue culture seedlings
Soaking the tissue culture seedlings for 2 days by using a nutrient solution formula of international rice, and then cleaning the tissue culture seedlings by using natural water at the temperature of 20-25 ℃, so as to remove the nutrient solution at the roots of the tissue culture seedlings and prevent the root systems of the tissue culture seedlings from mildewing after transplanting;
2) hormone treatment
Placing the cleaned tissue culture seedling into a test tube, adding natural water containing abscisic acid to submerge the root, culturing for 1-2 days under the condition of shade in a greenhouse, spraying gibberellin solution on the leaf surface of the seedling before transplanting, keeping the temperature and shading by using a black gauze;
3) soil preparation
Filling 7-10 kg of dry soil in each barrel of paddy field soil, adding water after filling the soil, mashing the soil uniformly, and standing the soil in a greenhouse for 2 days so as to balance soil heating and floating soil precipitation;
4) transplantation and environmental management
Transplanting, namely transplanting the seeds singly, wherein the transplanting depth is proper for the soil to just cover the roots of the seedlings, irrigating 1-2 cm of shallow water after transplanting the seedlings, shading the seedlings for one week by using a black gauze, and then removing the black gauze to culture the seedlings under the natural illumination of a greenhouse;
5) liquid manure management
Irrigating shallow water 4 weeks after transplanting, applying 10-15 rice compound fertilizers, naturally drying, maintaining a dry-wet alternating soil environment, and applying fertilizer for one week to plant in the field;
the content of abscisic acid in the natural water containing abscisic acid is 10-100 mg/L, the concentration of the gibberellin solution is 1-10 mg/L, and the spraying condition is that 2-5 ml of gibberellin solution is sprayed on the leaf surface of each seedling.
2. The culture method for improving the transplanting survival rate of the tissue culture seedlings of rice as claimed in claim 1, wherein the natural water is tap water exposed to the sun for 5 days.
3. The cultivation method for improving the transplanting survival rate of the rice tissue culture seedlings as claimed in claim 1, wherein the shading condition of the black gauze is 1/10 light intensity reaching normal light to avoid direct sunlight.
4. A culture method for improving the transplanting survival rate of tissue culture seedlings of rice as claimed in claim 1, wherein the transplanting and environmental management further comprises the steps of weeding once with shallow water after 15 days of transplanting, naturally drying after weeding until the soil has fine cracks, then watering, and then maintaining the dry-wet alternative soil environment.
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