CN110731268B

CN110731268B - Tissue culture and rapid propagation method for ficus carica

Info

Publication number: CN110731268B
Application number: CN201911057716.4A
Authority: CN
Inventors: 徐爱春; 章书声; 郭瑞; 孙骏威
Original assignee: China Jiliang University
Current assignee: China Jiliang University
Priority date: 2019-11-01
Filing date: 2019-11-01
Publication date: 2021-08-03
Anticipated expiration: 2039-11-01
Also published as: CN110731268A

Abstract

The invention discloses a tissue culture and rapid propagation method of ficus carica, which sequentially comprises the following steps: (1) and (3) disinfection and inoculation of the ficus carica stem sections: taking a new stem of the ficus carica extracted in the current year, shearing the stem into stem segments with axillary buds, and inoculating; (2) inducing cluster buds; (3) multiplication of adventitious buds; (4) rooting culture of the regenerated plant; (5) hardening and transplanting the seedlings. The culture mediums with different treatments, different components and proportions are adopted in the steps. By adopting the method, the henbane plants can be quickly obtained, and the industrial production is facilitated. By adopting the method, the adventitious bud induction rate of the cluster buds of the henbane fruits is as high as 86.7 percent, the number of the adventitious buds is 7.4, the multiplication multiple of the adventitious buds can reach 6.9, the rooting rate can reach 97 percent, and the plant transplanting survival rate can reach more than 98 percent.

Description

Tissue culture and rapid propagation method for ficus carica

Technical Field

The invention belongs to the technical field of plant culture, and particularly relates to a tissue culture and rapid propagation method of ficus carica.

Background

Ficus indica [ Ficus erecta thunb. var. beecheyana (hook. et ann.) King ] is small deciduous tree or shrub of Ficus genus of Moraceae (Moraceae), and produces Guangdong (and coastal island), Guangxi, Guizhou, Hubei (Wuhan and Shi Wei), Hunan, Jiangxi, Fujian, Zhejiang and Taiwan. Living under mountain slope forest or brook side. The fruit is used as a medicine named as Tianxiao fruit, also named as cow milk, milk pulp, wild loquat, Dayeniuzi, Maotaixianguo, milk bead, mountain milk, mountain fig and Dahao young milk, has the effects of relaxing bowel, detoxifying and reducing swelling, and is mainly used for treating symptoms such as constipation, hemorrhoid and gall; the root of the medicine is named as the root of milk plasm and also named as Dahao tieniu Diishan, has warm, sweet and pungent properties, has the efficacies of replenishing qi to invigorate the spleen, promoting blood circulation to remove meridian obstruction, dispelling wind and eliminating dampness and the like, and is mainly used for treating diseases such as fatigue and hypodynamia, anorexia, milk retention, spleen deficiency leucorrhea, rectocele, irregular menstruation, headache and pain, traumatic injury, rheumatic arthralgia and the like; the stem and leaf of the Chinese medicinal composition are used as medicines, namely the bupleurum root, the marsdenia tenacissima and the milk tea, have warm nature, sweet and light taste, have the effects of tonifying qi and strengthening spleen, dispelling wind and warm, and promoting blood circulation and removing obstruction in channels, and are mainly used for treating qi deficiency and hypodynamia, soreness and weakness of limbs, rheumatic arthralgia, unfavorable bones and muscles, traumatic injury, amenorrhea and galactostasis. At present, the ficus carica is basically in a wild state, and the ficus carica in the wild state is distributed in a scattered shape, which shows that the efficiency of breeding by seeds is not high. At present, no research report on rapid propagation of ficus carica exists.

At present, the research reports of tissue culture and rapid propagation of ficus plants are mostly concentrated on garden plants such as ficus elastica, linden, ficus elastica and the like and fruit trees such as figs and the like, used explants are mostly stem sections, stem tips and leaves, but the induction rate and the proliferation rate of adventitious buds are generally low, and a basic culture medium used for experiments generally adopts MS with high ion concentration. The Moraceae plant, especially Ficus plant, has a large amount of milk tubes distributed in stems and leaves, and the milk in the milk tubes contains a large amount of polyphenols, and is easy to flow out and be oxidized into brown quinones in the air during injury or in vitro culture, so as to cause browning of the culture medium, thereby influencing the smooth operation of tissue culture. And how to prevent browning is not substantially concerned in the literature.

Disclosure of Invention

The technical problem to be solved by the invention is as follows: aiming at the defects in the prior art, the tissue culture and rapid propagation method of the ficus tikoua, which has high survival rate and is beneficial to realizing industrialized production, is provided.

In order to realize the purpose of the invention, the following technical scheme is adopted for realizing the purpose: a method for tissue culture and rapid propagation of ficus carica comprises the following steps in sequence:

(1) disinfection of the stem section of ficus indica

Soaking the current-year stem of Ficus carica Linn in appropriate amount of liquid detergent for 2 min, washing with flowing water for 0.5-1.0 hr, draining, refrigerating at 4 deg.C for 0-48 hr, taking out, and soaking in superclean bench with 1 drop of Tween-80 and 0.1% of HgCl₂Soaking in the solution for 10-12 min, soaking in sterile water for 3 times, cutting head and tail of sterile filter paper, cutting into stem with axillary bud, soaking in 1% ascorbic acid solution for 0-60 min, and soaking in sterile water for 3 times to obtain sterilized explant material.

(2) Induction of clumpy buds

Inoculating the explant material obtained in the last step into a WPM culture medium according to growth polarity for culture, wherein the culture medium is added with ZT not more than 2.0 mg/L, 6-BA not more than 5.0 mg/L, PVP not more than 5.0 g/L, CH not more than 1.0 g/L, sucrose 30 g/L, and agar 9.0 g/L, and the pH value is 5.8-6.0; culturing the explant in dark for 1 day in culture mediumThe culture temperature is 21-25 ℃; then, the culture is transferred to illumination culture under the culture conditions that: the culture temperature is 21-25 deg.C, the illumination time is 14 hr/day, and the illumination intensity is 30-40 micromoles/(meter)²Seconds);

(3) proliferation of adventitious buds

Cutting the adventitious bud cluster obtained by the previous step into bud blocks with 2-3 adventitious buds, inoculating the bud blocks into WPM culture medium for culture, wherein the culture medium is added with NAA not more than 1.0 mg/L, 6-BA not more than 5.0 mg/L, CH not more than 1.0 mg/L, PVP not more than 5.0 g/L, sucrose 30 g/L and agar 9.0 g/L, and the pH value is 5.8-6.0. The culture conditions were: culturing at 21-25 deg.C, with illumination time of 14 hr/day and illumination intensity of 30-40 micromoles/m²Second;

(4) rooting culture of regenerated plants

Cutting adventitious buds with tender green leaves, vigorous growth and 3-4 leaves, inserting the adventitious buds into a WPM culture medium with the concentration of 12.5% -100% for rooting, wherein the culture medium is added with NAA not more than 1.0 mg/L, AC not more than 5.0 g/L, 20-30 g/L sucrose and 8.0 g/L agar, and the pH value is 5.8-6.0; the culture conditions were: culturing at 21-25 deg.C, with illumination time of 14 hr/day and illumination intensity of 30-40 micromoles/m²Second;

(5) hardening off and transplanting

The height of the tissue culture seedling to be rooted is more than 3 cm, when the root length is more than 5 cm, the bottle cap of the tissue culture bottle is loosened, a small amount of sterile water is injected into the bottle to prevent the culture medium from drying and cracking, the bottle cap is removed after 6 hours, the sterile water is added, the bottle cap is removed after 18 hours, and the seedling is grown at 30-40 micromoles/meter²Second light intensity of the light directly irradiated the regenerated plants for 2 days, during which sterile water was added several times. Then carefully triturating the culture medium, taking out the regenerated plants, carefully washing the residual agar with running water, planting the plants in the disinfected vegetable garden soil added with vermiculite, wherein the plant spacing is more than 2 cm, thoroughly watering, covering with a transparent polyethylene plastic film for heat preservation and moisture preservation, and controlling the indoor temperature to be 18-22 ℃. Removing four corners of the polyethylene plastic film after 3 days, completely removing the polyethylene plastic film the next day, and taking soil after the new leaves are unfoldedTransplanting to field.

As a preferable scheme: cleaning the stem section inoculated in the step (1), and then refrigerating the stem section at the low temperature of 4 ℃ for 24 hours in a refrigerator, wherein the stem section is subjected to HgCl reaction₂The solution was sterilized and washed and then soaked in 1% ascorbic acid solution for 20 minutes.

As a preferable scheme: the WPM minimal medium in the step (2) is added with 0.2-0.5 mg/L ZT, 2.0 mg/L6-BA, 0.2-0.5 g/L CH and 0.5-1.0 g/L PVP.

As a preferable scheme: in the step (3), the WPM minimal medium is added with 0.2 mg/L NAA, 1.0 mg/L6-BA, 0.2-0.5 g/L CH and 0.5-1.0 g/L PVP.

As a preferable scheme: the rooting medium formula in the step (4) is that 50% WPM is added with 0.1-0.2 mg/L NAA and 1.0-2.0 g/L AC.

As a preferable scheme: the volume ratio of the vermiculite added into the sterilized vegetable garden soil in the step (5) is 1/10-20.

Compared with the prior art, the invention has the beneficial effects that: the method of the invention can reduce browning as much as possible, improve multiplication coefficient, quickly obtain the henbane regenerated plant and is beneficial to industrialized production. By adopting the method, the induction rate of the cluster buds of the stem segments of the ficus carica is as high as 86.7 percent, the number of the adventitious buds is 7.4, the multiplication multiple of the adventitious buds can reach 6.9, the rooting rate can reach 100 percent, and the plant transplanting survival rate can reach more than 98 percent.

Detailed Description

Example 1

The abbreviations are annotated as: HgCl₂Mercuric chloride; AC activated carbon; 6-BA 6-benzylamino adenine; CH hydrolyzed casein; NAA naphthaleneacetic acid; ZT zeatin; PVP polyvinylpyrrolidone.

A method for tissue culture and rapid propagation of ficus carica comprises the following steps in sequence:

(1) disinfection of the stem section of ficus indica

Soaking stem of Ficus carica Linn in appropriate amount of liquid detergent for 2 min, washing with flowing water for 0.5-1.0 hr, draining, refrigerating at 4 deg.C for 24 hr, taking out, and refrigerating at ultra-high temperatureThe clean bench is dipped and added with 1 drop of Tween-80 and HgCl with the mass fraction of 0.1%₂Soaking in the solution for 10 min, then soaking in sterile water for 3 times, cutting head and tail of sterile filter paper, cutting into stem with axillary bud, soaking in 1% ascorbic acid solution for 20 min, and soaking in sterile water for 3 times to obtain sterilized explant material. The browning rate counted 20 days after inoculation was lower than 5%.

(2) Induction of clumpy buds

The explant material obtained in the previous step was inoculated to a WPM minimal medium containing ZT 0.5 mg/L, 6-BA 2.0 mg/L, PVP 1.0 g/L, and CH0.5 g/L, wherein sucrose was added in an amount of 30 g/L, agar was added in an amount of 9.0 g/L, pH was 5.8-6.0, and the medium was sterilized at 121 ℃ for 20 minutes, as follows. Culturing the culture medium inoculated with the explant in the dark for 1 day at the temperature of (23 +/-2) DEG C; then, the culture is transferred to illumination culture under the culture conditions that: culturing at 21-25 deg.C, with illumination time of 14 hr/day and illumination intensity of 30-40 micromoles/m²Second; axillary buds began to germinate at day 8, cluster buds began to be visible at day 12, and the number of adventitious buds for each explant was found to be 7.4 at day 30. No browning of the medium occurred.

(3) Proliferation of adventitious buds

Cutting the adventitious bud cluster obtained by the previous step into bud blocks with 2-3 adventitious buds, inoculating the bud blocks into a WPM basic culture medium added with NAA 0.2 mg/L, 6-BA1.0 mg/L, CH0.5 g/L and PVP 1.0 g/L for culture under the culture conditions that: culturing at 21-25 deg.C, with illumination time of 14 hr/day and illumination intensity of 30-40 micromoles/m²Second; after 30 days, the multiplication times of the adventitious buds are counted to be 6.9. No browning of the medium occurred.

(4) Rooting culture of regenerated plants

Cutting adventitious buds of 3-4 leaves which are tender green and vigorous in growth, inserting the buds into a 50% WPM basic culture medium which is added with NAA0.1 mg/L and AC 1.0 g/L for rooting, wherein the amount of added cane sugar is 20 g/L, the amount of added agar is 8.0 g/L, the pH value is 5.8-6.0, and sterilizing the buds at 121 ℃ for 20 minutes; the rooting starts in 12 days, and the rooting rate can reach 100 percent in 25 days.

(5) Hardening off and transplanting

When the height of the tissue culture seedling to be rooted is more than 3 cm and the root length is more than 5 cm, loosening the bottle cap of the tissue culture bottle, injecting a small amount of sterile water into the bottle to prevent the culture medium from drying and cracking, moving the bottle cap after 6 hours, adding sterile water, taking 18 hours, removing the bottle cap, and allowing the seedling to grow to 30-40 micromoles/meter²Second light intensity of the light directly irradiated the regenerated plants for 2 days, during which sterile water was added several times. Then carefully triturating the culture medium, taking out the regenerated plants, carefully washing the residual agar with running water, and planting the plants in the sterilized vegetable garden soil added with vermiculite according to the volume ratio of 1: 20, the plant spacing is more than 2 cm, the plant is thoroughly watered and covered by a transparent polyethylene plastic film for heat preservation and moisture preservation, and the indoor temperature is controlled between 18 and 22 ℃. After 3 days, the four corners of the polyethylene plastic film are uncovered, the polyethylene plastic film is completely uncovered the next day, and the leaves can be transplanted to the field with soil after being unfolded, and the transplanting survival rate can reach more than 98 percent.

Example 2

The present example differs from example 1 in step (1): sterilizing the fruit stems of the garden balsam: soaking the current-year stem of Ficus carica Linn in appropriate amount of liquid detergent for 2 min, washing with flowing water for 0.5-1.0 hr, draining, refrigerating at 4 deg.C for 0-48 hr, taking out, and soaking in superclean bench with 1 drop of Tween-80 and 0.1% of HgCl₂Soaking in the solution for 10 min, washing with sterile water for 3 times, cutting head and tail of sterile filter paper, cutting into stem with axillary bud, soaking in 1% ascorbic acid solution for 0-60 min, and soaking in sterile water for 3 times to obtain sterilized explant material. Temperature changes may modulate enzyme activity, whereas ascorbic acid is an antioxidant, both treatments may have a mitigating effect on the occurrence of browning. As shown in Table 1, statistics of 30 days after inoculation revealed that the browning rate of the untreated explants inoculated in the medium with PVP was still 33.3%, but the browning rate was significantly reduced after the low temperature treatment or the 1% ascorbic acid soaking treatment, while the two treatments were performedThe combination of which has a superimposed effect. It can be seen that the treatment according to the claims (low temperature pretreatment for 24h, 1% ascorbic acid soaking time for 20 min) performed best.

TABLE 1 Effect of pretreatment on explant browning Rate and Cluster bud Induction

Example 3

The present example differs from example 1 in step (2): inoculating the explant material obtained in the last step into WPM culture medium according to growth polarity, and culturing, wherein the culture medium is added with ZT not more than 2.0 mg/L, 6-BA not more than 5.0 mg/L, PVP not more than 5.0 g/L, CH not more than 1.0 g/L, sucrose 30 g/L, agar 9.0 g/L, and pH value is 5.8-6.0. Culturing the culture medium inoculated with the explant in the dark for 1 day at 21-25 deg.C; then, the culture is transferred to illumination culture under the culture conditions that: the culture temperature is 23 + -2 deg.C, the illumination time is 14 hr/day, and the illumination intensity is 30-40 micromole/m²Second. As shown in Table 2, after 30 days of culture, the WPM medium added with ZT 0.5 mg/L, 6-BA 2.0 mg/L, PVP 1.0 g/L and CH0.5 g/L has the highest adventitious bud induction rate and adventitious bud number and the browning rate is also the lowest; when the 6-BA is used alone, the inductivity of the adventitious bud and the number of the adventitious buds are greatly reduced, so that the two cytokinins have synergistic action. Meanwhile, from the aspect of browning, the influence on the browning rate is obvious if PVP is added, because PVP is a specific adsorbent for polyphenol substances, substrates are lacked, enzymatic reaction is naturally reduced, CH is reported to reduce browning, and the effect is really played from the aspect of data although the effect is not obvious. ZT and 6-BA with too high concentration have great promotion effect on browning, and the obtained adventitious bud may also have vitrification phenomenon to influence subsequent culture. Due to browning, a decrease in the adventitious bud induction rate and a decrease in adventitious buds are caused. Therefore, the best performance is achieved by the treatment in the claims (ZT 0.5 mg/L, 6-BA 2.0 mg/L, PVP 1.0 g/L, CH0.5 g/L).

TABLE 2 Effect of different treatments on Cluster bud Induction

Example 4

The present example differs from example 1 in step (3): proliferation of adventitious buds: cutting the adventitious bud cluster obtained by the previous step into bud blocks with 2-3 adventitious buds, inoculating the bud blocks into a WPM culture medium for culture, wherein the culture medium is added with NAA not more than 1.0 mg/L, 6-BA not more than 5.0 mg/L, CH not more than 1.0 mg/L, PVP not more than 5.0 g/L, sucrose 30 g/L, agar 9.0 g/L and pH value is 5.8-6.0. The culture conditions were: culturing at 21-25 deg.C, with illumination time of 14 hr/day and illumination intensity of 30-40 micromoles/m²Second; as shown in Table 3, statistics shows after 30 days, the addition of PVP can substantially avoid browning, thereby indirectly improving the multiplication factor. The increase of the concentration of 6-BA improves the multiplication times, but the vitrification of stems and leaves is brought by overhigh concentration. Experience shows that addition of a proper amount of auxin in a multiplication culture medium is beneficial to subsequent rooting culture; CH has some promotion of fold proliferation because it can provide various amino acids. In summary, the claimed WPM minimal medium treated (NAA 0.2 mg/L, 6-BA1.0 mg/L, CH0.5 g/L, PVP 1.0 g/L) is best suited for adventitious bud proliferation.

TABLE 3 Effect of different treatments on adventitious bud proliferation

Culture medium additive	Fold of proliferation	Growth conditions
			NAA 0.2，6-BA 1.0，CH 0.5，PVP 1.0	6.9	Thick stem and fast growth
NAA 0.2，6-BA 1.0，CH 0.5	5.5	Slow growth and browning of culture medium
			6-BA 2.0，CH 0.5，PVP 1.0	6.4	Thick stem and high growth speed
6-BA 2.0	5.8	Slow growth and browning of culture medium
			NAA 0.2，6-BA 0.2，CH 0.5，PVP 1.0	4.2	Fast growth and thick stem
NAA 1.0，6-BA 5.0，CH 0.5	3.5	Vitrification of stem and leaf and browning of culture medium

Example 5

The present example differs from example 1 in step (4): cutting adventitious buds with tender green leaves, vigorous growth and 3-4 leaves, inserting the adventitious buds into a WPM culture medium with the concentration of 12.5% -100% for rooting culture, wherein the culture medium is added with NAA not more than 1.0 mg/L, AC not more than 5.0 g/L, sucrose 20-30 g/L, agar 8.0 g/L and pH 5.8-6.0. As can be seen from Table 4, the rooting percentage is 55-100% and the rooting time is 12.3-19.4 days after 30 days of culture. The WPM minimal medium with the concentration of 50% without any hormone has the lowest rooting rate, but the rooting rate is obviously increased after NAA is added; the proper low-concentration WPM is beneficial to rooting of tissue culture seedlings, because the transformation of the tissue culture seedlings from heterotrophy to autotrophy can be promoted, but the shortage of nutrients is brought to the end of the culture seedlings, so that the stem leaves turn yellow; the influence of the NAA concentration on the growth of the rooting and tissue culture seedlings is that the rooting rate and the growth become stronger along with the increase of the concentration, but the rooting rate is reduced to some extent, the stem and the root are poor in performance, the stem and the leaf with over-high concentration are vitrified, the root is shortened and thickened, and the water and fertilizer absorption capacity is weakened; the AC can provide a dark environment for the culture medium, simulates the environment in soil, and can also play a role in preventing the culture medium from browning, but the high-concentration AC can also bring the defect of absorbing the nutrition of the culture medium. With the comprehensive table 4, the optimal combination is 50% WPM, 0.1 mg/L NAA, 2.0 g/L AC, 100% rooting rate, 5.6 cultured roots, moderate root thickness, strong stem and thick green seedling, and is suitable for hardening seedling and transplanting.

TABLE 4 Effect of different treatments on Cluster bud Induction

Example 6

The present example differs from example 1 in step (5): the height of the tissue culture seedling to be rooted is more than 3 cm, when the root length is more than 5 cm, the bottle cap of the tissue culture bottle is loosened, a small amount of sterile water is injected into the bottle to prevent the culture medium from drying and cracking, the bottle cap is removed after 6 hours, the sterile water is added, the bottle cap is removed after 18 hours, and the seedling is grown at 30-40 micromoles/meter²Second light intensity of the light directly irradiated the regenerated plants for 2 days, during which sterile water was added several times. Then carefully mashing the culture medium, taking out the regenerated plant, carefully washing the residual agar with running water, and planting the plant in the disinfected vegetable garden soil with vermiculite added, wherein the plant spacing is more than 2 cmAnd pouring water, covering with transparent polyethylene plastic film to keep temperature and moisture, and controlling indoor temperature at 18-22 deg.C. And 3 days later, uncovering four corners of the polyethylene plastic film, uncovering the polyethylene plastic film on the next day, and transplanting the film to the field with soil after the new leaves are unfolded. As can be seen from table 5, the addition of a proper amount of vermiculite to the vegetable garden soil can improve the survival rate of transplanting, because vermiculite can play a role in water and fertilizer conservation, but too much vermiculite does not improve the survival rate of transplanting, so the vermiculite is selected in consideration of the economy: the vegetable garden soil is 1: 20 in proportion.

TABLE 5 influence of different vermiculite addition on the survival rate of transplantation

Vermiculite: vegetable garden soil	Survival Rate of transplantation (%)
		1：100	85
1：20	98
		1：1	98
10：1	97

Claims

1. A tissue culture and rapid propagation method of ficus carica is characterized by sequentially comprising the following steps:

(1) disinfection of the stem section of ficus indica

Taking the current-year stem segment of the ficus carica, and adding a proper amount of detergentSoaking the refined solution for 2 min, washing with running water for 0.5-1.0 hr, draining, refrigerating in refrigerator at 4 deg.C for 24-48 hr, taking out, and dripping 1 drop of Tween-80 and HgCl 0.1 wt%₂Soaking and sterilizing in the solution for 10-12 min, then sufficiently soaking and washing with sterile water for 3 times, cutting off the head and the tail of the sterile filter paper, cutting into stem sections with axillary buds, soaking in a 1% ascorbic acid solution for 20-60 min, and sufficiently soaking and washing with sterile water for 3 times to obtain a sterilized explant material;

(2) induction of clumpy buds

Inoculating the explant material obtained in the last step to a culture medium WPM according to growth polarity, wherein the culture medium is added with 0.5 mg/L ZT, 2.0 mg/L6-BA, 1.0 g/L PVP, 0.5 g/L CH, 30 g/L sucrose, 9.0 g/L agar, and the pH value is 5.8-6.0; culturing the culture medium inoculated with the explant in the dark for 1 day at 21-25 deg.C; then, the culture is transferred to illumination culture under the culture conditions that: culturing at 21-25 deg.C, with illumination time of 14 hr/day and illumination intensity of 30-40 micromoles/m²Second;

(3) proliferation of adventitious buds

Cutting the adventitious bud cluster obtained by the previous step into bud blocks with 2-3 adventitious buds, inoculating the bud blocks into a culture medium WPM for culture, wherein the culture medium is added with 0.2 mg/L NAA, 1.0 mg/L6-BA, 0.5 g/L CH, 1.0 g/L PVP, 30 g/L sucrose and 9.0 g/L agar, and the pH value is 5.8-6.0; the culture conditions were: culturing at 21-25 deg.C, with illumination time of 14 hr/day and illumination intensity of 30-40 micromoles/m²Second;

(4) rooting culture of regenerated plants

Cutting adventitious buds with tender green leaves, vigorous growth and 3-4 leaves, inserting the adventitious buds into a culture medium WPM with the concentration of 12.5% -100% for rooting, wherein the culture medium is added with 0.1 mg/L NAA, 1.0 g/L AC, 20 g/L sucrose and 8.0 g/L agar, the pH value is 5.8-6.0, and the culture conditions are as follows: culturing at 21-25 deg.C, with illumination time of 14 hr/day and illumination intensity of 30-40 micromoles/m²Second;

(5) hardening off and transplanting

The height of the tissue culture seedling to be rooted is more than 3 cm, when the root length is more than 5 cm, the bottle cap of the tissue culture bottle is loosened, a small amount of sterile water is injected into the bottle to prevent the culture medium from drying and cracking, the bottle cap is removed after 6 hours, the sterile water is added, the bottle cap is removed after 18 hours, and the seedling is grown at 30-40 micromoles/m²Directly irradiating the regenerated plant with light of second light intensity for 2 days, adding sterile water for a plurality of times, carefully mashing the culture medium, taking out the regenerated plant, carefully washing the residual agar with running water, planting the plant in the disinfected vegetable garden soil with vermiculite added, wherein the plant spacing is more than 2 cm, watering thoroughly, covering with a transparent polyethylene plastic film for heat preservation and moisture preservation, controlling the indoor temperature at 18-22 ℃, uncovering four corners of the polyethylene plastic film after 3 days, uncovering all the polyethylene plastic film after the next day, and transplanting the plant to the field with soil after new leaves are uncovered.

2. The method for tissue culture and rapid propagation of ficus carica according to claim 1, wherein the method comprises the following steps: the stem segments in the step (1) are refrigerated for 24 hours at a low temperature of 4 ℃ in a refrigerator after being cleaned and are subjected to HgCl₂The solution was sterilized and washed and then soaked in 1% ascorbic acid solution for 20 minutes.

3. The method for tissue culture and rapid propagation of ficus carica according to claim 1, wherein the method comprises the following steps: the culture medium WPM in the step (2) is added with 0.2-0.5 mg/L ZT, 2.0 mg/L6-BA, 0.2-0.5 g/L CH and 0.5-1.0 g/L PVP.

4. The method for tissue culture and rapid propagation of ficus carica according to claim 1, wherein the method comprises the following steps: in the step (3), the culture medium WPM is added with 0.2 mg/L NAA, 1.0 mg/L6-BA, 0.2-0.5 g/L CH and 0.5-1.0 g/L PVP.

5. The method for tissue culture and rapid propagation of ficus carica according to claim 1, wherein the method comprises the following steps: the WPM with the culture medium concentration of 50% for rooting culture in the step (4) is added with NAA of 0.1-0.2 mg/L and AC of 1.0-2.0 g/L.

6. The method for tissue culture and rapid propagation of ficus carica as claimed in claim 1, wherein the method comprises the following steps: the volume ratio of vermiculite added into the sterilized vegetable garden soil in the step (5) is 1: 10 to 20.