CN110402817A - A kind of method of rooting outside the bottle of emerald bamboo tissue culture seedling - Google Patents

Abstract

The invention discloses an out-of-bottle rooting method of emerald bamboo tissue culture seedlings, which comprises the steps of preparation of rooting facilities, acquisition of rooting materials, treatment of rooting materials, transplanting of tissue cultured seedlings, management of the rooting period, and management of the hardening period. The invention simulates the rooting environment of green bamboo test-tube seedlings in a bottle, so that the tissue-cultured seedlings can directly take root in a relatively closed, high-humidity, outer-bottle substrate with rooting liquid, and then gradually reduce the air humidity to enter the field stage of substrate cultivation. The method combines the process of rooting the tissue cultured seedlings in the agar medium in the bottle and then transplanting to the greenhouse, so that the tissue cultured seedlings can directly take root outside the bottle and in the transplanting medium, which simplifies the production process, saves production time, and reduces production cost. The rooting rate outside the bottle is more than 95%, and the survival rate of transplanting is 90%, which provides technical support for the large-scale and industrial production of green bamboo tissue culture seedlings.

Description

A kind of method of rooting outside the bottle of emerald bamboo tissue culture seedling

technical field

The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for rooting out of a bottle of emerald bamboo tissue culture seedlings.

Background technique

In recent years, with the rapid development of science and technology, plant tissue culture technology has entered the stage of production and application, and is widely used in large-scale in vitro rapid propagation of flowers, trees, and bamboo. The rapid propagation of plant tissue culture seedlings can be produced all year round under artificial control conditions, and a large number of tissue culture seedlings can be obtained in a short period of time.

Emerald bamboo is a small ground-cover bamboo species with dense branches and leaves and a huge underground root system. It is a good ground-cover bamboo species and water and soil conservation bamboo species. Therefore, the current application demand is increasing, and its economic value is also increasing. The higher the value, the brighter the market prospect.

The bamboo tissue culture work in my country began in the 1990s, using the tender joints of two species of Bamboo bamboo and Indian bamboo as explants to induce callus, and finally formed a complete plant (Que Guoning et al., 1991). The tissue culture research on bud propagation has been carried out on bamboo species such as emerald bamboo (Zhang Chunxia et al., 2010), Philippine white bamboo (Zhang Chunxia et al., 2006), pagoda, and spicy leek bamboo (Wang Guangping et al., 2005). The bamboo tissue culture research report is a study on the tissue culture technology of Yunnan Tianlong (Zhong Yongli et al., 2019), and most of the above form complete plants. At present, the technology of realizing the regeneration of bamboo plants through tissue culture has become mature, but the bamboo tissue culture seedlings are transplanted and rooted in a special medium under sterile conditions, and the production cost is high; Planting, from the weak light, constant temperature, near-saturated humidity, and sterile environment in the bottle to the natural outdoor environment, the requirements for the production environment are high, and the temperature and humidity control is difficult, which restricts the scale and industrialization of bamboo tissue culture seedlings. bottleneck in production. All current reports on the rooting technology of emerald green bamboo tissue culture seedlings are all carried out in the bottle, and the research on the rooting of bamboo tissue culture seedlings to reach bamboo plant regeneration has not yet been reported by the rooting technology outside the bottle.

Contents of the invention

In view of the deficiencies in the above-mentioned prior art, the technical problem to be solved by the present invention is to provide a method for rooting bamboo tissue culture seedlings outside the bottle, through the rooting technology outside the bottle to achieve bamboo plant regeneration, the original in the tissue culture room The two steps of transplanting the rooted bottle seedlings to the greenhouse for hardening are combined into one link: transplant the unrooted tissue cultured seedlings directly to the greenhouse for rooting, and the rooting rate is above 95%. At the same time, the seedling hardening is completed, and the survival rate reaches more than 90%, which not only greatly reduces the cost of tissue culture seedling production, but also shortens the production time. Moreover, the production facilities and equipment are simple and convenient, the operation is simple, and the germination rate is high.

In order to solve the problems of the technologies described above, the technical solution adopted in the present invention is:

A method for rooting out of a bottle of emerald green bamboo tissue culture seedlings, comprising the steps of preparation of rooting facilities, acquisition of rooting materials, processing of rooting materials, transplanting of tissue cultured seedlings, management of rooting period, and management of hardening period. The specific steps are:

1) Put a matrix mixed with nutrient soil, vermiculite and perlite in the hole of the plug, make the surface of the matrix flat and flush with the top edge of the plug, and use a mixture of 1000 times chlorothalonil and 1000 times carbendazim Soak the matrix for disinfection, and spray again before official use;

2) Transport the emerald green bamboo subculture bottle seedlings that have been subcultured for more than 20-30 days into the greenhouse, take out the clump-like sprouts from the medium with tweezers, and thoroughly wash the medium at the base of the clump of seedlings with running water ;

3) Immerse the cleaned tissue-cultured sprout cluster base in a container equipped with rooting liquid, and seal the film so that the sprout cluster is in a closed space;

4) Divide the sprout clusters soaked in rooting liquid into clusters of 10-30 sprouts, and transplant them into hole trays. The sprout clusters are planted as they are divided, and the rooting liquid is poured again immediately after planting. As rooting water, carry out rooting culture;

5) In the rooting culture stage after the transplanting of the tissue cultured sprouts, the temperature in the greenhouse is controlled between 20-30°C, 70% sunshade nets are used in the greenhouse for shade, and the humidity is kept above 90%;

6) After rooting and culturing for 18 days, 3-5 roots will grow from the base of the seedling cluster, and when the root length reaches 1-3 cm, transplant the rooting seedling cluster from the hole tray to the nutrition bag containing soil and decomposed organic fertilizer together with the original substrate , pour water thoroughly, use 70% shade net in the greenhouse for shade, spray 4 times a day, spray 30 seconds once, spray once in the morning, evening, before noon and after noon, and spray 2 times a day after 10 days, spray 30 seconds once, Spray once in the morning and in the afternoon to make the seedlings gradually adapt to the open field growth environment. After 10 days, move them into the field with soil and manage them according to the method of raising seedlings in the field.

In step 1), the length, width and height of the hole plate are 54 cm, 28 cm, and 11 cm respectively, and the number of holes is 4×8.

In step 1), the volume ratio of nutrient soil, vermiculite and perlite is 3:3:1.

In step 3), the rooting solution is 1.0mg/L indolebutyric acid IBA+1.5mg/L naphthaleneacetic acid NAA solution.

In step 3), the rooting solution is a commercially available rooting agent formulated with a concentration of 1g/L.

In step 3), the base of the sprouts enters the rooting solution for 2 cm and soaks for 8 hours.

In step 4), the depth of planting the base of the sprout cluster into the substrate is 1-1.5 cm.

In step 5), the humidity is controlled by an automatic gap spray device, spraying for 20 seconds every 15 minutes.

Step 6) The mass ratio of middle soil to decomposed organic fertilizer is 3:1.

In step 6), the nutrition bag is a degradable non-woven fabric with a length, width and height of 8cm, 8cm and 12cm respectively.

Beneficial effect: compared with the prior art, the present invention has the following technical advantages:

1) The present invention simulates the rooting environment in the bottle of emerald green bamboo test tube seedlings, so that the tissue cultured seedlings directly take root in a relatively closed, high-humidity, bottle-external substrate with rooting liquid, and then gradually reduce the air humidity to enter the field stage of substrate cultivation. The method combines the process of rooting the tissue cultured seedlings in the agar medium in the bottle and then transplanting to the greenhouse, so that the tissue cultured seedlings can directly take root outside the bottle and in the transplanting medium, which simplifies the production process, saves production time, and reduces production cost. The technical method is simple, the production cost is low, and the seedling rate is high, which is reflected in direct economic benefits in production.

2) The present invention can not only simplify the rooting process in the bottle, but also improve the survival rate of transplanting, and the growth condition is good, thereby shortening the time for group cultivation of seedlings, accelerating the propagation speed of seedlings, reducing the cultivation space, and simplifying the production process.

Description of drawings

Fig. 1 is a rooting material: no root emerald green bamboo tissue culture bud cluster figure;

Fig. 2 is the picture of the day when rootless emerald green bamboo tissue culture bud clusters are transplanted to plug trays;

Fig. 3 is the rooting status figure when rootless emerald green bamboo tissue culture seedling cluster is transplanted to hole tray 18 days;

Fig. 4 is that the emerald bamboo seedling clump is transplanted to the root growth status figure after 20 days of growth of the non-woven nutrition bag;

Fig. 5 is the root growth status diagram after the emerald green bamboo seedling cluster is transplanted to the non-woven nutrition bag growth 20 days; Among the figure, left: the rooting material is the tissue culture bud seedling cluster of transfer subcultivation 30 days; Right: the rooting material is transfer Subculture the tissue-cultured sprout clusters for 20 days;

Fig. 6 is the root growth state diagram after the emerald bamboo seedling cluster is transplanted to non-woven nutrition bag growth 20 days; Among the figure, left: the rooting material is the tissue culture bud seedling cluster that 30 buds are a cluster; Right: the rooting material is A group of tissue-cultured sprouts with 10 buds as a cluster.

Detailed ways

The present invention will be further described below in conjunction with specific examples, but these examples are not used to limit the present invention.

Example 1

A method for rooting out of a bottle of emerald green bamboo tissue culture seedlings, comprising the following steps:

1) Preparation of rooting facilities outside the bottle: the number of holes is 4 × 8, and the hole of the hole plate with a length of 54 cm, a width of 28 cm and a depth of 11 cm is filled with nutrient soil, vermiculite and perlite in a ratio of 3: 3: 1. The mixed matrix makes the surface of the matrix flat and flush with the top edge of the tray. Soak the matrix with 1000 times chlorothalonil and 1000 times carbendazim mixture for disinfection, and spray again before official use;

2) Acquisition of rooting material: transport the green bamboo subculture bottle seedlings that have been subcultured for 20 days into the greenhouse, take out the clump-like sprouts (as shown in Figure 1 ) from the culture medium with tweezers, and rinse them thoroughly with running water. Wash the culture medium at the base of the seedlings;

3) the processing of rooting material: the sprout cluster that cleans is put into container, pours rooting liquid in container, and rooting liquid is the solution of 1.0mg/L indolebutyric acid IBA+1.5mg/L naphthaleneacetic acid NAA, So that the base of the seedlings is completely immersed in the rooting solution for 2cm, and the container mouth is covered with a white plastic film, so that the sprouts are in a relatively closed space, and placed in the greenhouse for 8 hours for later use. The temperature of the greenhouse is controlled at 20-30°C. between.

4) Transplanting of tissue-cultured seedlings: Divide the sprout clusters soaked in the rooting liquid into about 30 sprouts, and transplant them into the hole trays. 1-1.5 centimeters, the hole tray container that will be planted full of seedlings is neatly placed in the solar greenhouse (as shown in Figure 2), pours the rooting liquid again as the fixed root water, carries out rooting cultivation;

5) Rooting stage management: In the stage of rooting cultivation after the transplanting of the group cultured sprout clusters, the temperature in the greenhouse is controlled between 20-30°C, 70% of the sunshade net is used for shading in the greenhouse, and an automatic gap spraying device is adopted, 15 minutes, spray for 20 seconds, keep the humidity above 90%;

6) Management during seedling hardening: After 18 days of rooting cultivation, 3-5 roots will grow from the base of the seedling cluster, and when the root length reaches 1-3 cm, transplant the rooting seedling cluster from the hole tray together with the original substrate to a place containing soil and decomposed organic matter. In the nutrition bag of the fertilizer, water it thoroughly in time, and the ratio of soil to decomposed organic fertilizer is 3:1. Use 70% shading nets in the greenhouse for shading. Spray 4 times a day, 30 seconds each time, once in the morning, evening, before noon and after noon. After 10 days, spray 2 times a day, 30 seconds once in the morning and after afternoon. Spray once each to make the seedlings gradually adapt to the open field growth environment. After another 10 days, they can be moved into the field with bags and soil, and managed according to the method of raising seedlings in the field.

Example 2

A method for rooting out of a bottle of emerald green bamboo tissue culture seedlings, comprising the following steps:

1) Preparation of rooting facilities outside the bottle: the number of holes is 4 × 8, and the holes are 54 cm long, 28 cm wide, and 11 cm deep. The formed matrix makes the surface of the matrix flat and flush with the upper edge of the plug. Soak the matrix with 1000 times chlorothalonil and 1000 times carbendazim mixture for disinfection, and spray again before official use;

2) Acquisition of rooting materials: Transport the bottle seedlings of emerald green bamboo subcultured for 30 days into the greenhouse, take out the clump-like sprouts from the medium with tweezers, and wash the clump base of the seedlings thoroughly with running water. base;

3) the processing of rooting material: the sprout cluster that cleans is put into container, pours rooting liquid in container, and rooting liquid is the solution of 1.0mg/L indolebutyric acid IBA+1.5mg/L naphthaleneacetic acid NAA, So that the base of the seedlings is completely immersed in the rooting solution for 2cm, and the container mouth is covered with a white plastic film, so that the sprouts are in a relatively closed space, and placed in the greenhouse for 8 hours for later use. The temperature of the greenhouse is controlled at 20-30°C. between.

4) Transplanting of tissue-cultured seedlings: Divide the sprout clusters soaked in the rooting liquid into about 30 sprouts, and transplant them into the hole trays. 1-1.5 centimeters; put the hole tray containers full of seedlings neatly in the solar greenhouse, and then pour the rooting water containing the rooting liquid again to carry out rooting culture;

5) Rooting stage management: In the stage of rooting cultivation after the transplanting of the group cultured sprout clusters, the temperature in the greenhouse is controlled between 20-30°C, 70% of the sunshade net is used for shading in the greenhouse, and an automatic gap spraying device is adopted, 15 minutes, spray for 20 seconds, keep the humidity above 90%;

6) Management during seedling hardening: After 18 days of rooting cultivation, 3-5 roots will grow from the base of the seedling cluster, and when the root length reaches 1-3 cm, transplant the rooting seedling cluster from the hole tray together with the original substrate to a place containing soil and decomposed organic matter. In the nutrition bag of the fertilizer, water it thoroughly in time, and the ratio of soil to decomposed organic fertilizer is 3:1. Use 70% shading nets in the greenhouse for shading. Spray 4 times a day, 30 seconds each time, once in the morning, evening, before noon and after noon. After 10 days, spray 2 times a day, 30 seconds once in the morning and after afternoon. Spray once each to make the seedlings gradually adapt to the open field growth environment. After another 10 days, they can be moved into the field with bags and soil, and managed according to the method of raising seedlings in the field.

Example 3

A method for rooting out of a bottle of emerald green bamboo tissue culture seedlings, comprising the following steps:

1) Preparation of rooting facilities outside the bottle: the number of holes is 4 × 8, and the holes are 54 cm long, 28 cm wide, and 11 cm deep. The formed matrix makes the surface of the matrix flat and flush with the upper edge of the plug. Soak the matrix with 1000 times chlorothalonil and 1000 times carbendazim mixture for disinfection, and spray again before official use;

2) Acquisition of rooting material: transport the subcultured tissue-cultured seedlings of emerald green bamboo for 30 days into the greenhouse, take out the clump-shaped sprouts from the medium with tweezers, and thoroughly wash the medium at the base of the clump of seedlings with running water;

3) the processing of rooting material: the sprout cluster that cleans is put into container, pours rooting liquid in container, and rooting liquid is 1.0mg/L indolebutyric acid IBA+1.5mg/L naphthaleneacetic acid NAA solution, makes The base of the seedlings is completely immersed in the rooting solution for 2 cm, and the container mouth is covered with a white plastic film, so that the sprouts are in a relatively closed space, and placed in the greenhouse for 8 hours for later use. The temperature of the greenhouse is controlled between 20-30 °C .

4) Transplanting of tissue-cultured seedlings: Divide the sprout clusters soaked in the rooting liquid into about 10 sprouts, and transplant them into the hole trays. 1-1.5 centimeters; place the tray containers full of seedlings neatly in the solar greenhouse, and then pour the rooting liquid again as rooting water for rooting cultivation;

5) Rooting stage management: In the stage of rooting cultivation after the transplanting of the group cultured sprout clusters, the temperature in the greenhouse is controlled between 20-30°C, 70% of the sunshade net is used for shading in the greenhouse, and an automatic gap spraying device is adopted, 15 minutes, spray for 20 seconds, keep the humidity above 90%;

6) Management during seedling hardening: After rooting and culturing for 18 days, observe the rooting situation. When 3-5 roots grow from the base of the seedling cluster and the root length reaches 1-3 cm, transplant the rooting seedling cluster from the plug to the In the nutrition bag containing soil and decomposed organic fertilizer, water it thoroughly in time, and the ratio of soil and decomposed organic fertilizer is 3:1. Use 70% shade nets in the greenhouse for shading. Spray 4 times a day, 30 seconds each time, once in the morning, evening, before noon and after noon. After 10 days, spray 2 times a day, 30 seconds once in the morning and after afternoon. Spray once each to make the seedlings gradually adapt to the open field growth environment. After another 10 days, they can be moved into the field with bags and soil, and managed according to the method of raising seedlings in the field.

Example 4

A method for rooting out of a bottle of emerald green bamboo tissue culture seedlings, comprising the following steps:

1) Preparation of rooting facilities outside the bottle: the number of holes is 4 × 8, and the holes are 54 cm long, 28 cm wide, and 11 cm deep. The formed matrix makes the surface of the matrix flat and flush with the upper edge of the plug. Soak the matrix with 1000 times chlorothalonil and 1000 times carbendazim mixture for disinfection, and spray again before official use;

2) Acquisition of rooting material: transport the subcultured tissue-cultured seedlings of emerald green bamboo for 30 days into the greenhouse, take out the clump-shaped sprouts from the medium with tweezers, and thoroughly wash the medium at the base of the clump of seedlings with running water;

3) Treatment of rooting materials: put the washed sprout clusters into a container, pour rooting liquid into the container, and the rooting liquid is prepared from a commercially available rooting agent (Beijing Aibi Biotechnology Co., Ltd., Shuangjier-GGR) Make a solution with a concentration of 1g/L, so that the base of the seedlings is completely immersed in the rooting solution for 2cm, and cover the mouth of the container with a white plastic film, so that the sprouts are in a relatively closed space, and put them in the greenhouse for 8 hours for later use. The greenhouse temperature is controlled between 20-30°C.

4) Transplanting of tissue-cultured seedlings: divide the sprout clusters soaked in the rooting liquid into 30 sprouts as a cluster, and transplant them into hole trays. The sprout clusters are planted as they are divided. 1-1.5 cm; Place the tray container full of seedlings neatly in the solar greenhouse, pour the rooting liquid again as the rooting water, and carry out rooting culture;

5) Rooting stage management: In the stage of rooting cultivation after the transplanting of the group cultured sprout clusters, the temperature in the greenhouse is controlled between 20-30°C, 70% of the sunshade net is used for shading in the greenhouse, and an automatic gap spraying device is adopted, 15 minutes, spray for 20 seconds, keep the humidity above 90%;

6) Management during seedling hardening: After rooting and cultivating for 18 days, observe the rooting situation. When 3-5 roots grow from the base of the seedling cluster and the root length reaches 5-8 cm, transplant the rooting seedling cluster from the plug to the In the nutrition bag containing soil and decomposed organic fertilizer, water it thoroughly in time, and the ratio of soil and decomposed organic fertilizer is 3:1. Use 70% shading nets in the greenhouse for shading. Spray 4 times a day, 30 seconds each time, once in the morning, evening, before noon and after noon. After 10 days, spray 2 times a day, 30 seconds once in the morning and after afternoon. Spray once each to make the seedlings gradually adapt to the open field growth environment. After 10 days, they can be moved into the field with bags and soil, and managed according to the method of raising seedlings in the field.

Before moving into the field, the rooting rate, root length, root system quantity of the emerald bamboo of the above-mentioned embodiment were measured and counted, and one month after moving into the field, the seedling rate was investigated and counted, and the measurement and statistics results are as shown in table 1, The emerald green bamboo rooting rate, root length, root system quantity, seedling rate of embodiment 2 are all higher than embodiment 1. Reason is that the number of days of subculture of the rooting material used in embodiment 1 is different from that of embodiment 2, which are respectively subcultured emerald green bamboo subcultured tissue culture seedlings of 20 days and 30 days. The days of subculture of the rooting material of embodiment 2 are longer, and the shoot cluster has more time for growth, and the shoot cluster is slightly higher and stronger than the shoot cluster of embodiment 1, so that the rooting ability is better than that of embodiment 1 Sprout clusters (Figure 5). The rooting rate of embodiment 3 and embodiment 2, root length, seedling rate are all similar, but the root system quantity of embodiment 3 single clump is about 1/3 of embodiment 2. This is because the sprout quantity that embodiment 3 transplants has only 1/3 that embodiment 2 causes.

Each embodiment rooting situation comparative table of table 1 (taking clump as unit to carry out statistics)

It should be noted that the above examples are only some specific embodiments of the present invention. Obviously, the present invention is not limited to the above embodiments, and many variations are possible. All deformations that can be directly derived or associated by those skilled in the art from the content disclosed in the present invention should be considered as the protection scope of the present invention.

Claims (10)

1. a method for taking root out of a bottle of emerald green bamboo tissue culture seedling, it is characterized in that, comprises the preparation of rooting facility, the acquisition of rooting material, the processing of rooting material, the transplanting of tissue culture seedling, rooting period management, hardening period management Steps, the specific steps are: 1) Put a matrix mixed with nutrient soil, vermiculite and perlite in the hole of the plug, make the surface of the matrix flat and flush with the top edge of the plug, and use a mixture of 1000 times chlorothalonil and 1000 times carbendazim Soak the matrix for disinfection, and spray again before official use; 2) Transport the emerald green bamboo subculture bottle seedlings that have been subcultured for 20-30 days into the greenhouse, take out the clump-shaped sprouts from the culture medium with tweezers, and thoroughly wash the culture medium at the base of the seedling clumps with running water; 3) Immerse the cleaned tissue-cultured sprout cluster base in a container equipped with rooting liquid, and seal the film so that the sprout cluster is in a closed space; 4) Divide the sprout clusters soaked in rooting liquid into clusters of 10-30 sprouts, and transplant them into hole trays. The sprout clusters are planted as they are divided, and the rooting liquid is poured again immediately after planting. As rooting water, carry out rooting culture; 5) In the rooting culture stage after the transplanting of the tissue cultured sprouts, the temperature in the greenhouse is controlled between 20-30°C, 70% sunshade nets are used in the greenhouse for shade, and the humidity is kept above 90%; 6) After rooting and culturing for 18 days, observe the rooting situation. When 3-5 roots grow from the base of the seedling cluster and the root length reaches 1-3 cm, transplant the rooting seedling cluster from the hole tray together with the original substrate to a place containing soil and decomposed organic matter. In the nutrition bag of the fertilizer, pour water thoroughly, use 70% sunshade net in the greenhouse for shade, spray 4 times a day, spray 30 seconds once, spray once in the morning, evening, before noon and after noon, and spray twice a day after 10 days. Spray once for 30 seconds, once in the morning and once in the afternoon, so that the seedlings gradually adapt to the open field growth environment. After another 10 days, move them into the field with bags and soil, and manage them according to the method of raising seedlings in the field. 2. the bottle rooting method of green bamboo tissue culture seedling according to claim 1, is characterized in that, in step 1), the volume ratio of nutrient soil, vermiculite, perlite is 3: 3: 1. 3. the bottle rooting method of emerald green bamboo tissue culture seedling according to claim 1, it is characterized in that, in step 1), the length, width and height of hole plate are respectively 54 centimetres, 28 centimetres, 11 centimetres, and number of holes is 4 ×8. 4. the rooting method outside the bottle of emerald green bamboo tissue culture seedling according to claim 1, it is characterized in that, in step 3), rooting liquid is 1.0mg/L indole butyric acid IBA+1.5mg/L naphthalene acetic acid NAA solution . 5. the rooting method outside the bottle of emerald green bamboo tissue culture seedling according to claim 1, it is characterized in that, in step 3), rooting liquid is that commercially available rooting agent is mixed with the solution that concentration is 1g/L. 6. the rooting method outside the bottle of the emerald green bamboo tissue culture seedling described in claim 1, it is characterized in that, in step 3), the sprout base is immersed in rooting liquid 2cm, soaks 8 hours. 7. the rooting method outside the bottle of the emerald green bamboo tissue culture seedling described in claim 1, it is characterized in that, in step 4), the bud seedling clump base is planted into matrix depth and is 1-1.5 centimetre. 8. the rooting method outside the bottle of emerald green bamboo tissue culture seedling according to claim 1, it is characterized in that, in step 5), humidity adopts automatic gap spraying device to control, every 15 minutes, spray 20 seconds. 9. the method for taking root outside the bottle of emerald green bamboo tissue culture seedling according to claim 1, it is characterized in that, the mass ratio of soil and decomposed organic fertilizer in step 6) is 3: 1. 10. The rooting method outside the bottle of emerald green bamboo tissue culture seedling according to claim 1, is characterized in that: in step 6), described nutrition bag is degradable non-woven fabric, and length, width, height are respectively 8cm, 8cm , 12cm.