CN112868529A - Method for improving seedling survival rate of hybrid filial generation of populus nigra - Google Patents
Method for improving seedling survival rate of hybrid filial generation of populus nigra 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
- A01H4/008—Methods for regeneration to complete plants
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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
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
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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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
- A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
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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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
- A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
- A01G24/15—Calcined rock, e.g. perlite, vermiculite or clay aggregates
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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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
- A01G24/28—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing peat, moss or sphagnum
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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
- A01G31/00—Soilless cultivation, e.g. hydroponics
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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
- A01G9/00—Cultivation in receptacles, forcing-frames or greenhouses; Edging for beds, lawn or the like
- A01G9/02—Receptacles, e.g. flower-pots or boxes; Glasses for cultivating flowers
- A01G9/029—Receptacles for seedlings
- A01G9/0291—Planting receptacles specially adapted for remaining in the soil after planting
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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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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures
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Abstract
The invention discloses a method for improving the seedling survival rate of hybrid filial generation of black poplar, which comprises the following steps: 1) after pollination for 30-40 days, the capsule is cut from the base part of the carpopodium, cleaned and disinfected, and the surface moisture of the capsule is sucked by sterile filter paper; 2) peeling off the peel along the longitudinal axis direction of the capsule core skin, and peeling off the catkin and the seed embryo; 3) inoculating the floccule and the seed embryo on an 1/2MS culture medium to be cultured until the height of the seedling is more than 4-6 cm and 4-6 true leaves exist; 4) washing the tissue culture seedling to remove the culture medium, transplanting the tissue culture seedling into a non-woven fabric container filled with the matrix, and transplanting the tissue culture seedling into a field after three weeks. The method is simple and feasible, safe and reliable, and the survival rate of the transplanted seedling in the field reaches more than 95 percent.
Description
Technical Field
The invention belongs to the technical field of plant seedling raising, and particularly relates to a method for improving the seedling raising survival rate of hybrid filial generation of populus nigra.
Background
The plant tissue culture is also called in vitro culture, and the in vitro organs (such as roots, stems, leaves, stem tips, flowers, fruits and the like), tissues or cells and protoplasts of the plant body are utilized to induce callus, adventitious buds and adventitious roots under the artificial conditions of sterility, a proper artificial culture medium, temperature and the like, and finally, a complete plant is formed. Crossbreeding is a main means for breeding new varieties of poplar. In order to improve the breeding efficiency and solve the problems of cluster shedding and immature seed development in the later stage of indoor branch water culture hybridization, the in vitro culture of the hybrid embryo in the later stage of water culture is a feasible and effective method. Tissue culture seedlings are obtained through in vitro culture, hardening and transplanting are carried out, and finally hybrid seedlings are obtained.
In the traditional tissue culture seedling transplanting, the tissue culture seedling is firstly transplanted into a substrate, hardening is finished indoors, and then the tissue culture seedling is transplanted into a field. The method has the advantages that the requirement on sterility, ventilation and nutrition required by the growth of the tissue culture seedling root is not easily met during indoor seedling hardening-out, the root of the tissue culture seedling is easily damaged during the process from indoor seedling hardening-out to field transplanting, the seedling revival period is longer, and the seedling survival rate of the hybrid progeny tissue culture seedling is low.
Disclosure of Invention
The invention aims to provide a method for improving the seedling survival rate of hybrid filial generation of populus nigra, and aims to solve the following problems:
1) the problems that the cutting branches of the black poplar drop off in the later stage of water culture hybridization, the seeds are immature and the field sowing and seedling formation are difficult;
2) when the hybrid offspring tissue culture seedling is acclimatized indoors, the substrate proportion is difficult to meet the requirements of sterility, ventilation and nutrition required by the growth of the tissue culture seedling root;
3) the roots of the tissue culture seedlings are easy to damage when the tissue culture seedlings are transplanted from indoor to field, the seedling revival period is longer, and the seedling rate of the tissue culture seedlings is low.
In order to solve the above problems, the present invention is specifically realized by the following technical scheme:
a method for improving the seedling survival rate of hybrid breeding filial generation of populus nigra comprises the following steps:
1) after pollination is carried out for 30-40 days, capsules are cut from the base parts of the fruit stalks, cleaned and disinfected, and the surface moisture of the capsules is sucked by sterile filter paper;
2) peeling off the seed coat along the longitudinal axis direction of the capsule core coat, and peeling off the catkin and the seed embryo;
3) inoculating the floccule and the seed embryo on an 1/2MS culture medium to be cultured until the height of the seedling is more than 4-6 cm and 4-6 true leaves exist;
4) putting the seedlings into clear water, removing the culture medium by using a brush to prevent the roots from being damaged, transplanting the seedlings into a nutrition cup filled with a matrix, and transplanting the seedlings into a field after 25-30 days.
Further, in the step (1), 75% alcohol is adopted for soaking and disinfection for 6-7 min. .
Further, in the step (3), the culture environment temperature is 25 +/-2 ℃, the culture environment temperature is 2500-3000 Lux, and the illumination time is 16-17 hours/day.
Further, the matrix in the step (4) comprises the following components in percentage by mass: 75% of turf, 15.5% of perlite, 7.5% of vermiculite, 1% of slow release fertilizer, 0.5% of calcium superphosphate and 0.5% of ferrous sulfate. The substrate container is made of non-woven fabrics, the specification is 5-7 cm in diameter and 10-12 cm in height, and the substrate with the thickness of 3-4 cm at the upper part of the container is replaced by river sand for sieving and disinfecting.
Further, the humidity of the environment transplanted to the nutrition cup in the step (4) from 1 st to 7 th days is kept at 95% -98%, the light transmittance is 75% -78%, and the temperature is 25 +/-2 ℃; keeping the humidity of the environment from 80% to 90% in 8 th to 15 th days; the light transmittance is 80-85%, and the temperature is 25 +/-2 ℃; keeping the humidity of the environment from 16 th to 23 th to 70 percent to 80 percent; the light transmittance is 90-95%, and the temperature is 25 +/-2 ℃; then spraying is carried out regularly according to the condition of the nursery stock, and the light transmittance is more than 95 percent.
Furthermore, the nutrition cup is made of non-woven fabrics and can be directly transplanted to a field, the seedling revival period and the root damage are reduced, and the transplanting survival rate is increased.
The invention has the beneficial effects that:
1) the problem that the cut branch water culture hybrid seeds of the black poplar are immature in development and difficult to sow in a field into seedlings is solved through tissue culture and seedling culture;
2) the root matrix of the tissue culture seedling is brushed away by a brush in clear water, so that the root damage is effectively reduced. A double-layer seedling raising matrix method of river sand and mixed matrix is adopted, and the requirements of sterility, ventilation and growth nutrition of hardening seedlings of tissue seedlings are met in a seedling raising container.
3) The nutrition cup is made of non-woven fabrics, the seedlings are directly transplanted in the nutrition cup, the seedlings are directly planted indoors after hardening, the seedling reviving period is not needed, and the transplanting survival rate is 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 invention provides a seedling hardening and transplanting method of tissue culture seedlings of poplar, which is used for improving the seedling survival rate of filial generation of cross breeding of poplar and is realized by the following steps:
1) and after pollination for 30 days, cutting capsules from the base parts of the fruit stalks by using scissors, washing the surfaces of the fruit peels by using tap water, draining water, and putting the fruit peels into a sterile bottle. Soaking and sterilizing with 75% alcohol for 6min, washing with sterile water for three times, placing the washed material into a culture dish, and drying the water on the surface of the material with sterile filter paper.
2) Lightly pressing along the longitudinal axis direction of the carpel by a blade to peel off the pericarp, lightly peeling off the catkin and the seed embryo, uniformly inoculating on 1/2MS culture medium, lightly pressing by an inoculating tool, and fully contacting the culture medium, so as to facilitate seedling emergence and growth. The culture environment temperature is 25 +/-2 ℃, the illumination intensity is 2000Lux, and the illumination time is 16h every day. The culture period is about 40 days, the height of a common seedling is more than 4cm, and 4-6 true leaves are provided.
3) The nutrition cup is prepared by using non-woven fabrics and light substrates as production raw materials. The air permeability, nutrients, etc. of the matrix play a critical role in inducing new roots.
The matrix formula comprises the following components in percentage by mass: 75% of turf, 15.5% of perlite, 7.5% of vermiculite, 1% of slow release fertilizer, 0.5% of calcium superphosphate and 0.5% of ferrous sulfate, and uniformly stirring. The non-woven fabric is made into a seedling container bag, and the seedling container bag is cut into nutrition cups by a machine, wherein the nutrition cups are 5.5cm in diameter and 10cm in height. Removing the matrix 3-4 cm from the upper part of the nutrition cup, filling the matrix into the screened and dried river sand, placing the nutrition cup in a seedling culture hole tray, and watering thoroughly for later use.
Before washing the seedlings, the water is dried to normal temperature in advance for later use. Removing the cover of the culture medium bottle seedling, rotating the culture medium bottle seedling in a culture bottle by using tweezers, pouring out the culture medium together with the seedling, removing a large culture medium, quickly immersing the whole seedling into a water basin, gently picking off poplar catkins at the base part of the seedling, gently brushing the culture medium on the root in a tray with water by using a fine hair brush, and keeping the root system complete as much as possible. And cleaning again, planting the seedlings into a prepared nutrition cup, watering thoroughly, enabling the seedlings and the matrix to be completely compact, keeping the tops and leaves of the seedlings clean, and avoiding adhering sand and the matrix. The humidity of the transplanting is kept about 95 percent in the first week, the humidity of the transplanting is kept between 85 and 90 percent in the second week, and the illumination condition is 3000 Lux. When the light is strong at noon, a shading net can be built. And observing the color of the sand and judging whether water is sprayed or not. After two weeks, at evening time or in cloudy day, seedlings with new roots and leaves are transplanted to an environment with humidity of about 70% and temperature of about 28 ℃. Reduce the humidity, further domesticate the seedlings, and adapt to the external environment.
4) Under the condition that the matrix is not lack of water, only a watering can is needed to spray water, so that the surface of the sand is ensured to be wet. After three weeks, selecting rainy days, transplanting the rooted seedlings into a field, building a thin shading net at the position 2.5m higher, and judging watering according to weather and matrix. After one week, the shading net on the arched shed is removed in rainy days, and after another week, the shading net is removed in rainy days, and the plants grow completely under natural conditions.
Example 2
The invention provides a seedling hardening and transplanting method for hybrid embryo tissue culture seedlings of Zhongxiang No. 3 Xjuba poplar, Danhong poplar Xzhong Han 17, Zhonghuo No. 6 Xjuba poplar and I-69 Xjuba poplar, which is used for improving the seedling survival rate of hybrid breeding progeny and is realized by the following contents:
1) pollinating pistil of populus davidiana 3, populus zhongzhu 6, populus I-69 and pollen of populus zhonghan 17 with pistil of populus davidiana respectively, shearing capsules from the base of fruit stalks by using scissors after 30 days, washing the surfaces of fruit peels by tap water, draining water, and putting into a sterile bottle. Soaking and sterilizing with 75% alcohol for 6min, washing with sterile water for three times, placing the washed material into a culture dish, and drying the water on the surface of the material with sterile filter paper.
2) Lightly pressing along the longitudinal axis direction of the carpel by a blade to peel off the pericarp, lightly peeling off the catkin and the seed embryo, uniformly inoculating on the MS culture medium, lightly pressing by an inoculating tool, and fully contacting the culture medium, thereby being beneficial to seedling emergence and growth. The culture environment temperature is 25 +/-2 ℃, the illumination intensity is 2000lx, and the illumination time is 16 hours per day. The culture period is about 40 days, the height of a common seedling is more than 4cm, and 4-6 true leaves are provided.
3) The nutrition cup is prepared by using non-woven fabrics and light substrates as production raw materials. The air permeability, nutrients, etc. of the matrix play a critical role in inducing new roots.
The matrix formula comprises the following components in percentage by mass: 75% of turf, 15.5% of perlite, 7.5% of vermiculite, 1% of slow release fertilizer, 0.5% of calcium superphosphate and 0.5% of ferrous sulfate, and uniformly stirring. The non-woven fabric is made into a seedling container, and the container is cut into sections by a machine to be made into nutrition cups, wherein the specification is that the diameter is 5.5cm and the height is 10 cm. Removing the matrix 4-5 cm from the upper part of the nutrition cup, filling the matrix into the screened and dried river sand, placing the nutrition cup in a seedling culture hole tray, and watering thoroughly for later use.
Before washing the seedlings, the water is dried to normal temperature in advance for later use. Removing the cover of the culture medium bottle seedling, rotating the culture medium bottle seedling in a culture bottle by using tweezers, pouring out the culture medium together with the seedling, removing a large culture medium, quickly immersing the whole seedling into a water basin, gently picking off poplar catkins at the base part of the seedling, gently brushing the culture medium on the root in a tray with water by using a fine hair brush, and keeping the root system complete as much as possible. And cleaning again, planting the seedlings into a prepared nutrition cup, watering thoroughly, enabling the seedlings and the matrix to be completely compact, keeping the tops and leaves of the seedlings clean, and avoiding adhering sand and the matrix. The humidity of the transplanting is kept about 95 percent in the first week, the humidity of the transplanting is kept between 85 and 90 percent in the second week, and the illumination condition is 3000 Lux. When the light is strong at noon, a shading net can be built. And observing the color of the sand and judging whether water is sprayed or not. After two weeks, at evening time or in cloudy day, seedlings with new roots and leaves are transplanted to an environment with humidity of about 70% and temperature of about 28 ℃. Reduce the humidity, further domesticate the seedlings, and adapt to the external environment.
4) Under the condition that the matrix is not lack of water, only a watering can is needed to spray water, so that the surface of the sand is ensured to be wet. After three weeks, selecting rainy days, transplanting the rooted seedlings into a field, building a thin shading net at the position 2.5m higher, and judging watering according to weather and matrix. After one week, the shading net on the arched shed is removed in rainy days, and after another week, the shading net is removed in rainy days, and the plants grow completely under natural conditions.
The seedling hardening transplantation and the seedling formation of control CK (direct sowing) of hybrid embryo tissue culture seedlings of Zhongxiong No. 3 XJuba poplar, Danhong poplar XZhonghan 17, Zhongzhu No. 6 XJuba poplar and I-69 XJuba poplar are shown in Table 1. As can be seen from the table, the method provided by the embodiment of the invention can increase the seedling rate of hybrid embryos of Zhongxiang No. 3 XJuba poplar, Danhong poplar XZhonghan 17, Zhonghuo No. 6 XJuba poplar and I-69 XJuba poplar by more than 95%, and improve the efficiency of poplar hybrid breeding.
TABLE 1 statistics of acclimatization and transplantation of tissue culture seedlings of hybrid poplar embryos and seedlings in control CK (direct sowing)
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 (5)
1. A method for improving the seedling survival rate of hybrid breeding offspring of populus nigra is characterized by comprising the following steps:
1) after pollination is carried out for 30-40 days, capsules are cut from the base parts of the fruit stalks, cleaned and disinfected, and the surface moisture of the capsules is sucked by sterile filter paper;
2) peeling off the seed coat along the longitudinal axis direction of the capsule core coat, and peeling off the catkin and the seed embryo;
3) inoculating the floccule and the seed embryo on an 1/2MS culture medium to be cultured until the height of the seedling is more than 4-6 cm and 4-6 true leaves exist;
4) washing the tissue culture seedling to remove the culture medium, transplanting the tissue culture seedling into a nutrition cup filled with the matrix, and transplanting the tissue culture seedling into a field after three weeks.
2. The method for improving seedling survival rate of hybrid populus nigra seedlings according to claim 1, wherein the disinfection in the step (1) is performed by soaking in 75% alcohol for 6-7 min.
3. The method for improving the seedling survival rate of hybrid populus nigra seedlings according to claim 1, wherein the cultivation environment temperature in the step (3) is 25 ℃ ± 2 ℃, the illumination intensity is 2500-3000 Lux, and the illumination time is 16-17 hours/day.
4. The method for improving seedling survival rate of hybrid populus nigra offspring according to claim 1, wherein the matrix in the step (4) comprises the following components in percentage by mass: 75% of turf, 15.5% of perlite, 7.5% of vermiculite, 1% of slow release fertilizer, 0.5% of calcium superphosphate and 0.5% of ferrous sulfate. The matrix container is made of non-woven fabrics, the diameter of the matrix container is 5-7 cm, the height of the matrix container is 10-12 cm, and matrix with the thickness of 3-4 cm on the upper portion of the container is replaced by river sand which is sieved and disinfected.
5. The method for improving the seedling survival rate of hybrid breeding of populus nigra according to claim 1, wherein the humidity of the environment transplanted to the nutrition cup from 1 to 7 days in the step (4) is kept at 95-98%, the light transmittance is 75-78%, and the temperature is 25 +/-2 ℃; keeping the humidity of the environment from 80% to 90% in 8 th to 15 th days; the light transmittance is 80-85%, and the temperature is 25 +/-2 ℃; keeping the humidity of the environment from 16 th to 23 th to 70 percent to 80 percent; the light transmittance is 90-95%, and the temperature is 25 +/-2 ℃; then spraying is carried out regularly according to the condition of the nursery stock, and the light transmittance is more than 95 percent.
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| CN115211359A (en) * | 2022-08-22 | 2022-10-21 | 中国林业科学研究院林业研究所 | Seedling hardening method for tissue culture seedlings of poplar |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115211359A (en) * | 2022-08-22 | 2022-10-21 | 中国林业科学研究院林业研究所 | Seedling hardening method for tissue culture seedlings of poplar |
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