CN110402817B - Method for rooting of green bamboo tissue culture seedlings outside bottles - Google Patents
Method for rooting of green bamboo tissue culture seedlings outside bottles Download PDFInfo
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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
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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
- 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
- A01G24/30—Growth substrates; Culture media; Apparatus or methods therefor based on or containing synthetic organic compounds
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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
Abstract
The invention discloses an ex-vitro rooting method for tissue culture seedlings of green bamboos, which comprises the steps of preparation of rooting facilities, acquisition of rooting materials, treatment of the rooting materials, transplantation of tissue culture plantlets, management of a rooting period and management of a seedling hardening period. The invention leads the tissue culture plantlet to directly root in a closed, high-humidity and bottle-shaped extracellular matrix with rooting liquid by simulating the rooting environment in the green bamboo test-tube plantlet bottle, then gradually reduces the air humidity and enters a matrix cultivation field stage. The method combines the process that the tissue culture seedling takes root in the agar culture medium in the bottle and then is transplanted to the greenhouse, so that the tissue culture seedling takes root directly outside the bottle and in the transplanting medium, thereby simplifying the production link, saving the production time and reducing the production cost. The ex-vitro rooting rate reaches more than 95 percent, the transplanting survival rate reaches 90 percent, and a technical support is provided for the large-scale and industrialized production of the green bamboo tissue culture seedlings.
Description
Technical Field
The invention belongs to the technical field of plant tissue culture, and particularly relates to a method for rooting of green bamboo tissue culture seedlings outside a bottle.
Background
In recent years, with the rapid development of science and technology, plant tissue culture technology has entered the production and application stage, and is widely applied to the large-scale in vitro rapid propagation of flowers, woods and bamboos. The rapid propagation of the plant tissue culture nursery stock can be produced all the year round under the condition of manual control, and a large amount of tissue culture nursery stock can be obtained in a short time.
The green bamboos are small ground cover bamboo species, the branches and leaves are dense, the underground rhizome system is huge, and the green bamboos are good ground cover greening bamboo species and water and soil conservation bamboo species, so that the current application demand is higher and higher, the economic value is higher and higher, and the green bamboos have wide market prospects.
Bamboo tissue culture work in China began in the 90's of the 20 th century, and callus was induced using tender nodes of yellow bamboo and Indian bamboo 2 rosette bamboo as explants to finally form whole plants (Jungonin et al, 1991). Tissue culture studies with bud propagation have been performed on bamboo species such as green bamboo (zucchini et al, 2010), fei-white bamboo (zucchini et al, 2006), plantain bamboo, and labeo-chive bamboo (wang guan-nu et al, 2005), and the most recent tissue culture studies on bamboo are tissue culture technical studies on yunnan saurolong (bellybus et al, 2019), which mostly form complete plants. At present, the technology for realizing the regeneration of bamboo plants through tissue culture tends to be mature, but the bamboo tissue culture seedlings are transferred and cultured to root in a special culture medium under the aseptic condition, so the production cost is high; the transplanting of the rooting seedlings from indoor to outdoor gradually changes from weak light, constant temperature, near saturated humidity and sterile environment in the bottle to outdoor natural environment conditions, has high requirements on production environment and large difficulty in temperature and humidity control, and is a bottleneck for restricting large-scale and industrialized production of the bamboo tissue culture seedlings. At present, all reports about rooting technologies of the green bamboo tissue culture seedlings are carried out in bottles, and researches for realizing rooting of the bamboo tissue culture seedlings to achieve regeneration of bamboo plants through an ex-bottle rooting technology are not reported.
Disclosure of Invention
Aiming at the defects in the prior art, the technical problem to be solved by the invention is to provide an out-of-bottle rooting method for the green bamboo tissue culture seedlings, which achieves the regeneration of bamboo plants through an out-of-bottle rooting technology, roots in a tissue culture seedling bottle which is originally carried out in a tissue culture room, transplants the rooted bottle seedlings to two links of greenhouse seedling hardening, and combines the two links into one link: the tissue culture seedlings which do not root are directly transplanted to a greenhouse for rooting, the rooting rate is over 95 percent, the seedling hardening is completed at the same time, and the survival rate is over 90 percent, so that the production cost of the tissue culture seedlings is greatly reduced, and the production time is shortened. And the production facility is simple and easy, the operation is simple, and the emergence rate is high.
In order to solve the technical problems, the invention adopts the technical scheme that:
an ex-vitro rooting method for tissue culture seedlings of green bamboos comprises the steps of preparation of rooting facilities, acquisition of rooting materials, treatment of the rooting materials, transplantation of tissue culture plantlets, management of a rooting period and management of a seedling hardening period, and specifically comprises the following steps:
1) filling a substrate mixed by nutrient soil, vermiculite and perlite into the hole tray, leveling the surface of the substrate and leveling the upper edge of the hole tray, soaking the substrate in 1000 times of chlorothalonil and 1000 times of carbendazim mixed solution for disinfection, and spraying again before formal use;
2) transporting the emerald green bamboo subculture bottle seedlings which are subjected to subculture for more than 20-30 days into a greenhouse, taking out cluster bud seedlings from the culture medium by using tweezers, and thoroughly washing the culture medium at the base of the seedling cluster by using running water;
3) immersing the base part of the cleaned tissue culture bud seedling cluster into a container filled with a rooting solution, and sealing the container by using a thin film to ensure that the bud seedling cluster is in a closed space;
4) dividing the bud seedling cluster subjected to the soaking treatment by the rooting solution into bud seedling clusters with 10-30 bud seedlings as a cluster, transplanting the bud seedling clusters into a plug tray, planting the bud seedling clusters along with the division, watering the rooting solution again as rooting solution immediately after planting, and carrying out rooting culture;
5) in the rooting culture stage after tissue culture bud seedling transplantation, the temperature in a greenhouse is controlled to be 20-30 ℃, a shading net with 70 percent is adopted in the greenhouse to shade, and the humidity is kept to be more than 90 percent;
6) after 18 days of rooting culture, 3-5 roots grow at the base of the seedling cluster, when the root length reaches 1-3 cm, transplanting the rooted seedling cluster together with the original matrix into a nutrition bag containing soil and decomposed organic fertilizer, watering, shading by adopting a 70% shading net in a greenhouse, spraying for 4 times every day, spraying for 30 seconds in the morning, evening, before and after noon, spraying for 2 times every day after 10 days, spraying for 30 seconds every day, and spraying for once after the morning and afternoon to ensure that the seedlings gradually adapt to an open field growing environment, after 10 days, transplanting the seedlings into a field with soil, and managing according to a field seedling culture method.
In the step 1), the length, width and height of the plug are respectively 54cm, 28cm and 11cm, and the number of the plugs is 4 multiplied by 8.
In the step 1), the volume ratio of the nutrient soil, the vermiculite and the perlite is 3: 1.
In the step 3), the rooting solution is a solution of 1.0mg/L of indolebutyric acid IBA and 1.5mg/L of naphthylacetic acid NAA.
In the step 3), the rooting solution is a solution prepared by a commercial rooting agent with the concentration of 1 g/L.
And in the step 3), the base part of the bud seedling enters 2cm of rooting liquid and is soaked for 8 hours.
In the step 4), the depth of the base of the bud seedling cluster planted in the substrate is 1-1.5 cm.
And 5), controlling the humidity by using an automatic gap spraying device, and spraying for 20 seconds every 15 minutes.
The mass ratio of the soil to the decomposed organic fertilizer in the step 6) is 3: 1.
In the step 6), the nutrition bag is made of degradable non-woven fabrics, and the length, width and height of the nutrition bag are respectively 8cm, 8cm and 12 cm.
Has the advantages that: compared with the prior art, the invention has the following technical advantages:
1) the invention simulates the rooting environment in the test-tube seedling bottle of the green bamboo, so that the tissue culture plantlet directly roots in the sealed and high-humidity bottle epimatrix with the rooting solution, then the air humidity is gradually reduced, and the culture plantlet enters the field culture stage of the matrix. The method combines the process that the tissue culture seedling takes root in the agar culture medium in the bottle and then is transplanted to the greenhouse, so that the tissue culture seedling takes root directly outside the bottle and in the transplanting medium, thereby simplifying the production link, saving the production time and reducing the production cost. The technical method is simple, low in production cost and high in seedling rate, and the direct economic benefit is realized in production.
2) The invention can simplify the links of rooting in the bottle, improve the survival rate of transplanting, and has good growth condition, thereby shortening the time of tissue culture seedling, accelerating the propagation speed of the nursery stock, reducing the culture space and simplifying the production links.
Drawings
FIG. 1 is a rooting material: a clump diagram of the rootless green bamboo tissue culture seedlings;
FIG. 2 is a current day diagram of clump transplanting of rootless Cuilus chinensis tissue culture seedlings to a plug tray;
FIG. 3 is a view showing the rooting condition of the unrooted green bamboo tissue culture seedlings when they are transplanted into the plug for 18 days;
FIG. 4 is a diagram showing the root growth status of the seedlings of the green bamboos after the seedlings are transplanted into a non-woven fabric nutrition bag and grow for 20 days;
FIG. 5 is a diagram showing the root growth after the green bamboo seedling cluster is transplanted into a non-woven fabric nutrition bag and grows for 20 days; in the figure, left: the rooting material is a tissue culture bud seedling cluster which is subjected to subculture for 30 days; and (3) right: the rooting material is a tissue culture bud seedling cluster which is subjected to subculture for 20 days;
FIG. 6 is a diagram showing the root growth after the green bamboo seedling cluster is transplanted into the non-woven fabric nutrition bag and grows for 20 days; in the figure, left: the rooting material is a tissue culture bud seedling cluster taking 30 buds as one cluster; and (3) right: the rooting material is a tissue culture bud seedling cluster taking 10 buds as one cluster.
Detailed Description
The present invention is further illustrated by the following specific examples, which are not intended to be limiting.
Example 1
An ex-vitro rooting method for green bamboo tissue culture seedlings comprises the following steps:
1) preparing an out-of-bottle rooting facility: filling a substrate mixed by nutrient soil, vermiculite and perlite according to the proportion of 3: 1 into the holes of a hole disk with the hole number of 4 multiplied by 8, the length of 54cm, the width of 28cm and the depth of 11cm, and enabling the surface of the substrate to be flat and level with the upper edge of the hole disk. Soaking the substrate with mixed solution of 1000 times of chlorothalonil and 1000 times of carbendazim for disinfection, and spraying again before formal use;
2) obtaining a rooting material: transferring the green bamboo subculture tissue culture bottle seedling subjected to subculture for 20 days into a greenhouse, taking out the cluster bud seedling from the culture medium by using tweezers (as shown in figure 1), and thoroughly cleaning the culture medium at the base of the seedling cluster by using running water;
3) treatment of rooting materials: putting the cleaned bud seedling cluster into a container, pouring a rooting solution into the container, wherein the rooting solution is a solution of 1.0mg/L indolebutyric acid IBA and 1.5mg/L naphthylacetic acid NAA, so that the base part of the seedling cluster is completely immersed into the rooting solution for 2cm, covering the opening of the container with a white plastic film, so that the bud seedling is in a closed space, putting the bud seedling cluster in a greenhouse for standing for 8 hours for later use, and controlling the temperature of the greenhouse to be between 20 and 30 ℃.
4) Transplanting the tissue culture plantlets: dividing the bud seedling cluster subjected to the soaking treatment by the rooting solution into about 30 bud seedlings as a cluster, transplanting the cluster into a plug tray, planting the bud seedling cluster along with the cluster, planting the base part of the bud seedling cluster into a substrate with the depth of 1-1.5 cm, neatly placing the plug tray container full of the seedlings in a sunlight greenhouse (as shown in figure 2), watering once again by the rooting solution as rooting water, and carrying out rooting culture;
5) and (3) management of a rooting period: in the rooting culture stage after the tissue culture bud seedling cluster is transplanted, the temperature in a greenhouse is controlled to be between 20 and 30 ℃, a 70 percent sunshade net is adopted to shade in the greenhouse, an automatic gap spraying device is adopted to spray for 20 seconds every 15 minutes, and the humidity is kept to be more than 90 percent;
6) and (3) seedling hardening period management: after rooting culture for 18 days, 3-5 roots grow at the base part of the seedling cluster, when the root length reaches 1-3 cm, the rooted seedling cluster is transplanted into a nutrition bag containing soil and decomposed organic fertilizer from a hole tray together with the original matrix, water is timely poured, and the ratio of the soil to the decomposed organic fertilizer is 3: 1. The greenhouse is shaded by adopting a 70% sunshade net, 4 times of spraying are carried out every day, 30 seconds of spraying are carried out once, spraying is carried out once in the morning, evening, before noon and after noon, spraying is carried out once after 10 days, 2 times of spraying is carried out every day, 30 seconds of spraying is carried out once, and spraying is carried out once in the morning and after afternoon, so that the nursery stocks are gradually adapted to the open field growth environment, and after 10 days, the nursery stocks can be moved into the field with the soil in bags and managed according to a field seedling culture method.
Example 2
An ex-vitro rooting method for green bamboo tissue culture seedlings comprises the following steps:
1) preparing an out-of-bottle rooting facility: filling a substrate mixed by nutrient soil, vermiculite and perlite according to the proportion of 3: 1 into the holes of a hole disk with the hole number of 4 multiplied by 8, the length of 54cm, the width of 28cm and the depth of 11cm, and enabling the surface of the substrate to be flat and level with the upper edge of the hole disk. Soaking the substrate with mixed solution of 1000 times of chlorothalonil and 1000 times of carbendazim for disinfection, and spraying again before formal use;
2) obtaining a rooting material: conveying the green bamboo subculture tissue culture bottle seedlings subjected to subculture for 30 days into a greenhouse, taking out cluster bud seedlings from the culture medium by using tweezers, and thoroughly cleaning the culture medium at the base of the seedling cluster by using running water;
3) treatment of rooting materials: putting the cleaned bud seedling cluster into a container, pouring a rooting solution into the container, wherein the rooting solution is a solution of 1.0mg/L indolebutyric acid IBA and 1.5mg/L naphthylacetic acid NAA, so that the base part of the seedling cluster is completely immersed into the rooting solution for 2cm, covering the opening of the container with a white plastic film, so that the bud seedling is in a closed space, putting the bud seedling cluster in a greenhouse for standing for 8 hours for later use, and controlling the temperature of the greenhouse to be between 20 and 30 ℃.
4) Transplanting the tissue culture plantlets: dividing the bud seedling cluster subjected to the soaking treatment by the rooting solution into about 30 bud seedlings as a cluster, transplanting the cluster into a plug tray, planting the bud seedling cluster along with the branch, and planting the base part of the bud seedling cluster into a substrate with the depth of 1-1.5 cm; the plug container full of the plantlets is placed in a sunlight greenhouse in order, and then rooting water containing rooting liquid is poured for the second time for rooting culture;
5) and (3) management of a rooting period: in the rooting culture stage after the tissue culture bud seedling cluster is transplanted, the temperature in a greenhouse is controlled to be between 20 and 30 ℃, a 70 percent sunshade net is adopted to shade in the greenhouse, an automatic gap spraying device is adopted to spray for 20 seconds every 15 minutes, and the humidity is kept to be more than 90 percent;
6) and (3) seedling hardening period management: after rooting culture for 18 days, 3-5 roots grow at the base part of the seedling cluster, when the root length reaches 1-3 cm, the rooted seedling cluster is transplanted into a nutrition bag containing soil and decomposed organic fertilizer from a hole tray together with the original matrix, water is timely poured, and the ratio of the soil to the decomposed organic fertilizer is 3: 1. The greenhouse is shaded by adopting a 70% sunshade net, 4 times of spraying are carried out every day, 30 seconds of spraying are carried out once, spraying is carried out once in the morning, evening, before noon and after noon, spraying is carried out once after 10 days, 2 times of spraying is carried out every day, 30 seconds of spraying is carried out once, and spraying is carried out once after the morning and afternoon, so that the nursery stocks are gradually adapted to the open field growth environment, and the nursery stocks can be moved into the field with the soil in bags after 10 days and are managed according to the field seedling culture method.
Example 3
An ex-vitro rooting method for green bamboo tissue culture seedlings comprises the following steps:
1) preparing an out-of-bottle rooting facility: filling a substrate mixed by nutrient soil, vermiculite and perlite according to the proportion of 3: 1 into the holes of a hole disk with the hole number of 4 multiplied by 8, the length of 54cm, the width of 28cm and the depth of 11cm, and enabling the surface of the substrate to be flat and level with the upper edge of the hole disk. Soaking the substrate with mixed solution of 1000 times of chlorothalonil and 1000 times of carbendazim for disinfection, and spraying again before formal use;
2) obtaining a rooting material: transferring the green bamboo subculture tissue culture seedlings subcultured for 30 days into a greenhouse, taking out the clumpy bud seedlings from the culture medium by using tweezers, and thoroughly cleaning the culture medium at the base of the seedling clumps by using running water;
3) treatment of rooting materials: putting the cleaned bud seedling cluster into a container, pouring a rooting solution into the container, wherein the rooting solution is a solution of 1.0mg/L of indolebutyric acid IBA and 1.5mg/L of naphthylacetic acid NAA, so that the base part of the seedling cluster is completely immersed into the rooting solution for 2cm, covering the opening of the container with a white plastic film, so that the bud seedling is in a closed space, putting the bud seedling cluster in a greenhouse for standing for 8 hours for later use, and controlling the temperature of the greenhouse to be between 20 and 30 ℃.
4) Transplanting the tissue culture plantlets: dividing the bud seedling cluster subjected to the soaking treatment by the rooting solution into about 10 bud seedlings as a cluster, transplanting the cluster into a plug tray, planting the bud seedling cluster along with the branch, and planting the base part of the bud seedling cluster into a substrate with the depth of 1-1.5 cm; the plug container full of the plantlets is placed in a sunlight greenhouse in order, and then the rooting solution is poured once again to serve as rooting water for rooting culture;
5) and (3) management of a rooting period: in the rooting culture stage after the tissue culture bud seedling cluster is transplanted, the temperature in a greenhouse is controlled to be between 20 and 30 ℃, a 70 percent sunshade net is adopted to shade in the greenhouse, an automatic gap spraying device is adopted to spray for 20 seconds every 15 minutes, and the humidity is kept to be more than 90 percent;
6) and (3) seedling hardening period management: and after rooting culture is carried out for 18 days, the rooting condition is observed, when 3-5 roots grow at the base part of the seedling cluster and the root length reaches 1-3 cm, the rooted seedling cluster is transplanted into a nutrition bag containing soil and decomposed organic fertilizer from a hole tray together with the original matrix, water is timely poured, and the ratio of the soil to the decomposed organic fertilizer is 3: 1. The greenhouse is shaded by adopting a 70% sunshade net, 4 times of spraying are carried out every day, 30 seconds of spraying are carried out once, spraying is carried out once in the morning, evening, before noon and after noon, spraying is carried out once after 10 days, 2 times of spraying is carried out every day, 30 seconds of spraying is carried out once, and spraying is carried out once after the morning and afternoon, so that the nursery stocks are gradually adapted to the open field growth environment, and the nursery stocks can be moved into the field with the soil in bags after 10 days and are managed according to the field seedling culture method.
Example 4
An ex-vitro rooting method for green bamboo tissue culture seedlings comprises the following steps:
1) preparing an out-of-bottle rooting facility: filling a substrate mixed by nutrient soil, vermiculite and perlite according to the proportion of 3: 1 into the holes of a hole disk with the hole number of 4 multiplied by 8, the length of 54cm, the width of 28cm and the depth of 11cm, and enabling the surface of the substrate to be flat and level with the upper edge of the hole disk. Soaking the substrate with mixed solution of 1000 times of chlorothalonil and 1000 times of carbendazim for disinfection, and spraying again before formal use;
2) obtaining a rooting material: transferring the green bamboo subculture tissue culture seedlings subcultured for 30 days into a greenhouse, taking out the clumpy bud seedlings from the culture medium by using tweezers, and thoroughly cleaning the culture medium at the base of the seedling clumps by using running water;
3) treatment of rooting materials: putting the cleaned bud seedling cluster into a container, pouring a rooting solution into the container, wherein the rooting solution is a commercially available rooting agent (Beijing Aibidi Biotechnology Co., Ltd., double Gill-GGR) and is prepared into a solution with the concentration of 1g/L, so that the base part of the seedling cluster is completely immersed into the rooting solution for 2cm, the mouth of the container is covered by a white plastic film, the bud seedling is positioned in a relatively closed space, and the container is placed in a greenhouse for standing for 8 hours for standby, and the temperature of the greenhouse is controlled to be between 20 and 30 ℃.
4) Transplanting the tissue culture plantlets: dividing the bud seedling cluster subjected to the soaking treatment by the rooting solution into 30 bud seedlings as a cluster, transplanting the cluster into a plug tray, planting the bud seedling cluster along with the branch, and planting the base part of the bud seedling cluster into a substrate with the depth of 1-1.5 cm; the plug container full of the plantlets is placed in a sunlight greenhouse in order, and then the rooting solution is poured once again to serve as rooting water for rooting culture;
5) and (3) management of a rooting period: in the rooting culture stage after the tissue culture bud seedling cluster is transplanted, the temperature in a greenhouse is controlled to be between 20 and 30 ℃, a 70 percent sunshade net is adopted to shade in the greenhouse, an automatic gap spraying device is adopted to spray for 20 seconds every 15 minutes, and the humidity is kept to be more than 90 percent;
6) and (3) seedling hardening period management: and after rooting culture is carried out for 18 days, the rooting condition is observed, when 3-5 roots grow at the base part of the seedling cluster and the root length reaches 5-8 cm, the rooted seedling cluster is transplanted into a nutrition bag containing soil and decomposed organic fertilizer from a hole tray together with the original matrix, water is timely poured, and the ratio of the soil to the decomposed organic fertilizer is 3: 1. The greenhouse is shaded by adopting a 70% sunshade net, 4 times of spraying are carried out every day, 30 seconds of spraying are carried out once, spraying is carried out once in the morning, evening, before noon and after noon, spraying is carried out once every day for 2 times after 10 days, spraying is carried out once for 30 seconds, and spraying is carried out once in the morning and after afternoon, so that the nursery stocks are gradually adapted to the open field growth environment, and after 10 days, the nursery stocks can be moved into the field with soil in bags and managed according to a field seedling culture method.
Before the green bamboos are transplanted into the field, the rooting rate, the root length, the number of root systems and the seedling rate of the green bamboos in the embodiment are measured and counted, the seedling rate is investigated and counted one month after the green bamboos are transplanted into the field, the measurement and counting results are shown in table 1, and the rooting rate, the root length, the number of root systems and the seedling rate of the green bamboos in the embodiment 2 are all higher than those in the embodiment 1. The reason is that the rooting materials used in example 1 and example 2 were subcultured for 20 days and 30 days, respectively, of the green bamboo subcultured tissue culture seedlings. The rooting material of example 2 was subcultured for a longer number of days, the shoot clump had more time for growth, and the shoot clump was slightly higher and stronger than the shoot clump of example 1, so that the rooting capacity was better than that of the shoot clump of example 1 (FIG. 5). The example 3 and the example 2 have similar rooting rate, root length and seedling rate, but the number of root systems of a single cluster in the example 3 is about 1/3 of the example 2. This is due to the fact that the number of sprouts transplanted in example 3 is only 1/3 of example 2.
TABLE 1 comparative rooting table for each example (statistics in clumps)
It is to be noted that the above-mentioned list is only a few specific embodiments of the present invention. It is obvious that the invention is not limited to the above embodiments, but that many variations are possible. All modifications which can be derived or suggested by a person skilled in the art from the disclosure of the present invention are to be considered within the scope of the invention.
Claims (10)
1. The ex-vitro rooting method for the tissue culture seedlings of the green bamboos is characterized by comprising the steps of preparing rooting facilities, obtaining rooting materials, treating the rooting materials, transplanting tissue culture plantlets, managing the rooting period and managing the seedling hardening period, and specifically comprises the following steps:
1) filling a substrate mixed by nutrient soil, vermiculite and perlite into the hole tray, leveling the surface of the substrate and leveling the upper edge of the hole tray, soaking the substrate in 1000 times of chlorothalonil and 1000 times of carbendazim mixed solution for disinfection, and spraying again before formal use;
2) transferring the green bamboo subculture tissue culture bottle seedlings which are subjected to subculture for 20-30 days into a greenhouse, taking out the cluster bud seedlings from the culture medium by using tweezers, and thoroughly cleaning the culture medium at the base of the seedling cluster by using running water;
3) immersing the base part of the cleaned tissue culture bud seedling cluster into a container filled with a rooting solution, and sealing the container by using a thin film to ensure that the bud seedling cluster is in a closed space;
4) dividing the bud seedling cluster subjected to the soaking treatment by the rooting solution into bud seedling clusters with 10-30 bud seedlings as a cluster, transplanting the bud seedling clusters into a plug tray, planting the bud seedling clusters along with the division, watering the rooting solution again as rooting solution immediately after planting, and carrying out rooting culture;
5) in the rooting culture stage after tissue culture bud seedling transplantation, the temperature in a greenhouse is controlled to be 20-30 ℃, a shading net with 70 percent is adopted in the greenhouse to shade, and the humidity is kept to be more than 90 percent;
6) and after 18 days of rooting culture, observing the rooting condition, transplanting 3-5 roots at the base of the seedling cluster into a nutrition bag containing soil and decomposed organic fertilizer together with the original matrix when the root length reaches 1-3 cm, watering thoroughly, shading by adopting a 70% shading net in a greenhouse, spraying for 4 times every day, spraying for 30 seconds once, spraying for 30 seconds in the morning, evening, before and after the noon, spraying for 2 times every day after 10 days, spraying for 30 seconds once, spraying for 30 seconds in the morning and after the afternoon, and spraying for one time after the morning and afternoon so that the seedlings gradually adapt to the open field growth environment, after 10 days, transplanting the seedlings into a field together with the bag soil, and managing according to a field seedling culture method.
2. The method for rooting outside the bottle of green bamboo tissue culture seedlings according to claim 1, wherein in the step 1), the volume ratio of the nutrient soil, the vermiculite and the perlite is 3: 1.
3. The method for rooting outside of green bamboo tissue culture seedlings according to claim 1, wherein in step 1), the length, width and height of the aperture disk are 54cm, 28cm and 11cm respectively, and the number of apertures is 4 x 8.
4. The ex vitro rooting method for green bamboo tissue culture seedlings according to claim 1, wherein in the step 3), the rooting solution is a 1.0mg/L solution of indolebutyric acid IBA +1.5mg/L solution of naphthaleneacetic acid NAA.
5. The ex vitro rooting method for green bamboo tissue culture seedlings according to claim 1, wherein in the step 3), the rooting solution is a solution prepared from a commercially available rooting agent with a concentration of 1 g/L.
6. The ex vitro rooting method for green bamboo tissue culture seedlings according to claim 1, wherein in step 3), the base of the seedling is immersed in a rooting solution for 2cm for 8 hours.
7. The ex vitro rooting method for green bamboo tissue culture seedlings according to claim 1, wherein in step 4), the depth of the base of the bud seedling cluster is planted in the substrate to be 1-1.5 cm.
8. The method for rooting outside green bamboo tissue culture seedlings outside a bottle as claimed in claim 1, wherein in step 5), the humidity is controlled by an automatic gap spraying device, and spraying is performed for 20 seconds every 15 minutes.
9. The ex vitro rooting method for green bamboo tissue culture seedlings according to claim 1, wherein the mass ratio of soil to decomposed organic fertilizer in step 6) is 3: 1.
10. The ex vitro rooting method for green bamboo tissue culture seedlings according to claim 1, characterized by comprising: in the step 6), the nutrition bag is made of degradable non-woven fabrics, and the length, width and height of the nutrition bag are respectively 8cm, 8cm and 12 cm.
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