CN108184670B - Method for tissue culture of monstera deliciosa - Google Patents
Method for tissue culture of monstera deliciosa Download PDFInfo
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- CN108184670B CN108184670B CN201810142422.0A CN201810142422A CN108184670B CN 108184670 B CN108184670 B CN 108184670B CN 201810142422 A CN201810142422 A CN 201810142422A CN 108184670 B CN108184670 B CN 108184670B
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- 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
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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
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Abstract
The invention provides a method for culturing monstera deliciosa tissue, which relates to the technical field of plant cultivation and propagation and comprises the following steps: pretreating explants, (II) culturing culture medium, (III) hardening off seedlings in a greenhouse, and (IV) transplanting rooted seedlings; the monstera deliciosa tissue culture method induces the callus of the cedar on MS culture media containing different hormone ratios, and through the culture of the induction culture media, the multiplication culture media and the rooting culture media, each culture medium provides factors required by explants in the rooting and seedling strengthening periods, good rooting conditions can be provided, healthy and strong seedlings are promoted, and the culture method is convenient in material obtaining, low in cost, simple to operate, high in propagation speed and high in propagation coefficient, has good economic benefits and ecological benefits, and is suitable for large-scale production.
Description
[ technical field ] A method for producing a semiconductor device
The invention relates to the technical field of plant cultivation and propagation, in particular to a method for tissue culture of monstera deliciosa.
[ background of the invention ]
Monstera deliciosa (school name: Monstera deliciosa). Also known as "Banana" or "Wiegia", evergreen vine. The stem is thick and strong; the young leaves have heart-shaped and no hole, and grow into wide egg shape and pinnate deep crack after growing up; the leaf veins have oval perforations, long handle and dark green color. Firework, Firework boat shape, 11 months flowering, light yellow. The stem is planted with brown air root which is shaped like an electric wire, so the stem is named as 'grass of electric wire'; the leaves are oval, many long round holes and deep cracks are distributed between pinnate leaf veins in a tortoise shell shape, the shape of the leaves is similar to a tortoise shell pattern, and the stems have nodes similar to bamboo trunks, so the leaves are named as 'monstera deliciosa'. The panicle inflorescence, the whole flower is like a 'desk lamp', and the whole flower is provided with a lampshade and a bulb. In the native Mexico tropical rainforest, the nature likes warm and humid environment, direct sunlight is forbidden, and the native Mexico tropical rainforest is not cold-resistant, and is used as indoor potted plant in the north of China.
In the cultivation process, the monstera deliciosa is usually propagated by seeds, the progeny variation is extremely large, the excellent characteristics of female parents can be maintained to the maximum extent through cutting propagation, tillering propagation and the like, the production cost is relatively low, along with the continuous improvement of the utilization level of the characteristic resources of the monstera deliciosa, particularly on the industrial poverty-deprivation project, the cutting propagation cannot meet the requirement of large-scale production in a short time, and the defects of undeveloped root systems, branch waste and the like are also caused, so that the research of the monstera deliciosa tissue culture technology for shortening the seedling culture time, improving the bud survival rate and meeting the large-scale production has very important significance.
[ summary of the invention ]
In view of the above, the present invention provides a method for culturing tortoise back bamboo tissue, the method for culturing tortoise back bamboo tissue induces callus of fir on MS culture medium containing different hormone ratios, each culture medium provides factors required by explants during rooting and seedling strengthening period through culture of the induction culture medium, the proliferation culture medium and the rooting culture medium, can provide good rooting conditions, and promote healthy and strong seedlings, and the culture method has the advantages of convenient material selection, low cost, simple operation, fast propagation speed, high propagation coefficient, good economic and ecological benefits, and is suitable for large-scale production.
In order to solve the technical problems, the invention adopts the following technical scheme:
a method for culturing monstera deliciosa tissue comprises the following steps:
firstly, pretreatment of explants: cutting the stem top or axillary buds of monstera deliciosa as an explant, cutting the explant into strips of 4-6cm, soaking the explant in 70% alcohol for 3-5s, soaking in 1-1.5% sodium hypochlorite solution for 15-30min, continuously oscillating in the soaking process, washing the explant with sterile water for 4-6 times after soaking, then placing the explant on a sterile inoculation tray to blow dry the surface moisture of the explant, cutting off leaves and old cuts, cutting into 2-2.5cm stem sections with 2-3 buds, placing the stem sections with buds under tap water to wash for 30min, and sucking the stem sections with buds with filter paper for later use;
(II) culture of the culture medium:
(1) and (3) induction culture: inoculating the stem section with the bud obtained in the step on an induction culture medium, culturing for 5-10 days under the dark condition, then alternately performing induction culture according to the ratio of light to dark being 2:1, cutting off the leaf of the bud after the bud grows to 4-5cm, and cutting into stem sections of 2-3 cm;
the induction culture medium contains MS +1.5mg/L KT +0.5 mg/L6-BA +0.1mg/L IBA +4.5g/L agar +30g/L sucrose;
(2) and (3) proliferation culture: inoculating the stem section obtained by the post-treatment in the step (1) to a proliferation culture medium for culturing for 35-40 days, and transferring the new bud to a new proliferation culture medium for culturing, wherein the proliferation multiple reaches 3-4 times;
the proliferation culture medium comprises the following components: MS +1.0mg/L KT +0.2 mg/L6-BA +0.5mg/L NAA +3.5g/L agar +25g/L sucrose;
(3) rooting culture: removing redundant lateral buds or adventitious buds of the cluster buds obtained by the enrichment culture in the step (2), cutting off small branches with single buds according to the height of 1.5-3cm, putting the cut small branches into a rooting culture medium for rooting culture, and taking out of a bottle after culturing for 20-25 days to obtain seedlings;
the rooting medium comprises the following components: 1/2 modified MS +0.1-1mg LIBA +1.5-2.0mg/L rooting powder +4-4.5g/L agar +25-30g/L white sugar;
(III) greenhouse seedling hardening: taking the plantlet out of the culture vessel by using tweezers, cleaning a culture medium at the root of the plantlet by using tap water, planting the plantlet into a culture soil seedling tray, spraying rooting water, spraying chlorothalonil with 1000 times of liquid of 800-.
In the invention, further, the tissue culture method further comprises a step of transplanting the rooted seedlings, which specifically comprises the following steps: hardening the seedlings for 20-30 days until the root system is changed from white to grey, transplanting the rooted seedlings, putting the rooted seedlings into clean water for rinsing when transplanting, cleaning the culture medium of the root, then soaking the rooted seedlings into a rooting agent for 5-10 minutes, and then transplanting the rooted seedlings into a mixed matrix container for culturing, wherein the mixed matrix is prepared by mixing the following components in a volume ratio of 1: 1-2: 2-3 of vermiculite powder, weathered coal and rice bran.
In the present invention, further, the conditions for inducing culture in step (1) are: the culture temperature is 23-27 deg.C, humidity is 68-82%, and pH is 5.6-6.0, dark culture is performed for 4-5 days, and light culture is performed, wherein the light culture intensity is 1500-.
In the present invention, the proliferation culture in step (2) is performed at a temperature of 25-30 deg.C, a humidity of 50-75%, and a pH of 5.6-6.0, wherein the light intensity of the proliferation culture is 1800-2200Lux, and the light time is 10-15 h/day.
In the invention, further, the rooting culture conditions in the step (3) are as follows: the illumination intensity is 2000-.
In the present invention, further, the modified MS in the step (3) includes 30 to 45 parts by weight of macronutrients, 0.3 to 0.6 parts by weight of micronutrients, and 5 to 10 parts by weight of organic reagents.
In the present invention, further, the macronutrient components and their corresponding concentrations are: 150mg/L of compound sodium nitrophenolate, 160mg/L of ammonium sulfate, 390mg/L of magnesium sulfate heptahydrate and 30-80mg/L of brassinosteroids.
In the present invention, further, the components of the micronutrients and their corresponding concentrations are: 15-18mg/L of manganese sulfate, 2-4mg/L of boric acid, 1-2mg/L of potassium iodide, 0.2-0.4mg/L of sodium molybdate, 0.05-0.08mg/L of cobalt chloride and 0.02-0.03mg/L of copper sulfate pentahydrate.
In the present invention, further, the components of the organic reagent and their corresponding concentrations are: 1-3mg/L of glycine, 1-3mg/L of nicotinic acid, 1-2mg/L of pyridoxine hydrochloride and 112mg/L of inositol 102-.
In summary, due to the adoption of the technical scheme, the invention has the beneficial effects that:
1. according to the method for tissue culture of monstera deliciosa, provided by the invention, the explant is sterilized before tissue culture, then the callus is induced on MS culture media containing different hormone ratios, and through culture of the induction culture media, the enrichment culture media and the rooting culture media, each culture medium provides factors required by the explant during rooting and seedling strengthening, so that good rooting conditions can be provided, healthy and strong seedlings can be promoted, and the regenerated seedlings of the fir can be obtained.
2. The method is improved on the basis of the traditional culture medium formula, some factors which can be needed in the process of rooting and seedling strengthening of the monstera deliciosa are added, callus is induced on the MS culture medium containing different hormone ratios, the MS culture medium is used as a basic culture medium, the components and the content of the monstera deliciosa rooting culture medium are improved, the culture conditions are adjusted, the transplanting survival rate of the monstera deliciosa can reach more than 90%, the seedlings grow robustly, and the economic value of the monstera deliciosa is enhanced.
[ detailed description ] embodiments
The following examples may help one skilled in the art to more fully understand the present invention, but are not intended to limit the invention in any way.
Example 1
The method for culturing the monstera deliciosa tissue provided by the embodiment comprises the following steps:
firstly, pretreatment of explants: cutting the stem top or axillary buds of monstera deliciosa as an explant, cutting the explant into 4cm strips, soaking the explant in 70% alcohol for 3s, soaking in 1% sodium hypochlorite solution for 15min, continuously oscillating in the soaking process, washing the explant with sterile water for 4 times after soaking, placing the explant on a sterile inoculation tray to blow dry the surface moisture of the explant, cutting off leaves and old cuts, cutting the explant into 2cm stem sections with 2 buds, placing the stem sections with buds under tap water for washing for 30min, and sucking the stem sections with buds with filter paper for later use;
(II) culture of the culture medium:
(1) and (3) induction culture: inoculating the stem section with the buds obtained in the step on an induction culture medium, culturing for 5 days under a dark condition, then alternately performing induction culture according to the ratio of light to dark time of 2:1, cutting leaves of the buds into stem sections of 2cm after the buds grow to 4 cm; wherein the induction culture medium contains MS +1.5mg/L KT +0.5 mg/L6-BA +0.1mg/L IBA +4.5g/L agar +30g/L sucrose; the conditions of the induction culture are as follows: culturing at 23 deg.C, humidity 68% and pH 5.6 in dark for 4 days, and culturing in light at illumination intensity of 1500lux for 12 hr per day;
(2) and (3) proliferation culture: inoculating the stem section obtained by the post-treatment in the step (1) to a proliferation culture medium for culturing for 35 days, and transferring the new bud to a new proliferation culture medium for culturing, wherein the proliferation multiple reaches 3 times; wherein, the components of the proliferation culture medium are as follows: MS +1.0mg/L KT +0.2 mg/L6-BA +0.5mg/L NAA +3.5g/L agar +25g/L sucrose; the proliferation culture is carried out under the conditions that the temperature is 25 ℃, the humidity is 50% and the pH value is 5.6, the illumination intensity of the proliferation culture is 1800Lux, and the illumination time is 10 h/day;
(3) rooting culture: removing redundant lateral buds or adventitious buds of the cluster buds obtained by the enrichment culture in the step (2), cutting off small branches with single buds according to the height of 1.5cm, putting the cut small branches into a rooting culture medium for rooting culture, and taking out of a bottle after culturing for 20 days and hardening seedlings; wherein, the components of the rooting culture medium are as follows: 1/2 modified MS +0.1mg LIBA +1.5mg/L rooting powder +4g/L agar +25g/L white sugar; the rooting culture conditions are as follows: the illumination intensity is 2000lx, the illumination time is 10 hours/d, and the temperature is 25 ℃; the modified MS comprises 30 parts by weight of macronutrient elements, 0.3 parts by weight of micronutrient elements and 5 parts by weight of organic reagent; wherein, the macronutrient elements have the following components and corresponding concentrations: 150mg/L of compound sodium nitrophenolate, 160mg/L of ammonium sulfate, 320mg/L of magnesium sulfate heptahydrate and 30mg/L of brassinosteroid; the micronutrient elements and their corresponding concentrations are: 15mg/L of manganese sulfate, 2mg/L of boric acid, 1mg/L of potassium iodide, 0.2mg/L of sodium molybdate, 0.05mg/L of cobalt chloride and 0.02mg/L of copper sulfate pentahydrate; the components of the organic reagent and their corresponding concentrations are: 1mg/L of glycine, 1mg/L of nicotinic acid, 1mg/L of pyridoxine hydrochloride and 102mg/L of inositol;
(III) greenhouse seedling hardening: taking the plantlets out of the culture vessel by using tweezers, cleaning culture media at the roots of the plantlets by using tap water, planting the plantlets in a culture soil seedling tray, spraying root fixing water, spraying chlorothalonil with 800 times of liquid after the leaves are dry, spraying 20 times of biogas liquid after 7 days, culturing in a greenhouse for 3 months, and then transplanting;
and (IV) transplanting rooted seedlings: after hardening the seedlings for 20 days until the root system is changed from white to grey, transplanting the rooted seedlings, putting the rooted seedlings into clean water for rinsing during transplanting, cleaning the culture medium of the root, then soaking the rooted seedlings in a rooting agent for 5 minutes, and then transplanting the rooted seedlings into a mixed matrix container for culturing, wherein the mixed matrix is prepared by mixing the following components in a volume ratio of 1: 1: 2, mixing the vermiculite powder, the weathered coal and the rice bran.
Example 2
The method for culturing the monstera deliciosa tissue provided by the embodiment comprises the following steps:
firstly, pretreatment of explants: cutting the stem top or axillary buds of monstera deliciosa as an explant, cutting the explant into strips of 5cm, soaking the explant in 70% alcohol for 4s, soaking in 1.2% sodium hypochlorite solution for 22min, continuously oscillating in the soaking process, washing the explant with sterile water for 5 times after soaking, then placing the explant on a sterile inoculation tray to blow dry the surface moisture of the explant, cutting off leaves and old cuts, cutting into stem sections of 2 buds of 2.2cm, placing the stem sections with buds under tap water for washing for 30min, and sucking the stem sections with buds with filter paper for later use;
(II) culture of the culture medium:
(1) and (3) induction culture: inoculating the stem section with the buds obtained in the step on an induction culture medium, culturing for 7 days under the dark condition, then alternately performing induction culture according to the ratio of light to dark time of 2:1, cutting off leaves of the buds after the buds grow to 4.5cm, and cutting into stem sections of 2.5 cm; wherein the induction culture medium contains MS +1.5mg/L KT +0.5 mg/L6-BA +0.1mg/L IBA +4.5g/L agar +30g/L sucrose; the conditions of the induction culture are as follows: culturing at 25 deg.C, humidity of 75% and pH of 5.8 in dark for 4 days, and culturing in light at light intensity of 1700lux for 13 hr per day;
(2) and (3) proliferation culture: inoculating the stem section obtained by the post-treatment in the step (1) to a proliferation culture medium for culturing for 37 days, and transferring the new bud to a new proliferation culture medium for culturing, wherein the proliferation multiple reaches 3.5 times; wherein, the components of the proliferation culture medium are as follows: MS +1.0mg/L KT +0.2 mg/L6-BA +0.5mg/L NAA +3.5g/L agar +25g/L sucrose; the proliferation culture is carried out under the conditions that the temperature is 27 ℃, the humidity is 62 percent and the pH value is 5.7, the illumination intensity of the proliferation culture is 2000Lux, and the illumination time is 12 h/day;
(3) rooting culture: removing redundant lateral buds or adventitious buds of the cluster buds obtained by the enrichment culture in the step (2), cutting off small branches with single buds according to the height of 2.2cm, putting the cut small branches into a rooting culture medium for rooting culture, and taking out of a bottle after the culture for 22 days and hardening seedlings; wherein, the components of the rooting culture medium are as follows: 1/2 modified MS +0.5mg LIBA +1.7mg/L rooting powder +4.2g/L agar +27g/L white sugar; the rooting culture conditions are as follows: the illumination intensity is 2250lx, the illumination time is 12 hours/d, and the temperature is 27 ℃; the improved MS comprises 30-45 parts by weight of macronutrient elements, 0.4 part by weight of micronutrient elements and 7 parts by weight of organic reagent; wherein, the macronutrient elements have the following components and corresponding concentrations: 170mg/L of compound sodium nitrophenolate, 180mg/L of ammonium sulfate, 360mg/L of magnesium sulfate heptahydrate and 55mg/L of brassinosteroid; the micronutrient elements and their corresponding concentrations are: 16mg/L of manganese sulfate, 3mg/L of boric acid, 1.5mg/L of potassium iodide, 0.3mg/L of sodium molybdate, 0.07mg/L of cobalt chloride and 0.03mg/L of copper sulfate pentahydrate; the components of the organic reagent and their corresponding concentrations are: 2mg/L of glycine, 2mg/L of nicotinic acid, 1.5mg/L of pyridoxine hydrochloride and 108mg/L of inositol;
(III) greenhouse seedling hardening: taking the plantlets out of the culture vessel by using tweezers, cleaning culture media at the roots of the plantlets by using tap water, planting the plantlets in a culture soil seedling tray, spraying root fixing water, spraying chlorothalonil with 900 times of liquid after the leaves are dry, spraying 25 times of biogas liquid after 11 days, culturing the plantlets in a greenhouse for 4 months, and transplanting;
and (IV) transplanting rooted seedlings: after hardening the seedlings for 25 days until the root system is changed from white to grey white, transplanting the rooted seedlings, putting the rooted seedlings into clean water for rinsing during transplanting, cleaning the culture medium of the root, then soaking the rooted seedlings in a rooting agent for 7 minutes, and then transplanting the rooted seedlings into a mixed matrix container for culturing, wherein the mixed matrix is prepared by mixing the following components in percentage by volume: 2: 2, mixing the vermiculite powder, the weathered coal and the rice bran.
Example 3
The method for culturing the monstera deliciosa tissue provided by the embodiment comprises the following steps:
firstly, pretreatment of explants: cutting the stem top or axillary buds of monstera deliciosa as an explant, cutting the explant into strips of 6cm, soaking the explant in 70% alcohol for 5s, soaking in 1.5% sodium hypochlorite solution for 30min, continuously oscillating in the soaking process, washing the explant with sterile water for 6 times after soaking, then placing the explant on a sterile inoculation tray to blow dry the surface moisture of the explant, cutting off leaves and old cuts, cutting into stem sections with 3 buds of 2.5cm, placing the stem sections with buds under tap water for washing for 30min, and sucking the stem sections with buds with filter paper for later use;
(II) culture of the culture medium:
(1) and (3) induction culture: inoculating the stem section with the buds obtained in the step on an induction culture medium, culturing for 10 days under a dark condition, then alternately performing induction culture according to the ratio of light to dark time of 2:1, cutting off leaves of the buds after the buds grow to 5cm, and cutting into stem sections of 3 cm; wherein the induction culture medium contains MS +1.5mg/L KT +0.5 mg/L6-BA +0.1mg/L IBA +4.5g/L agar +30g/L sucrose; the conditions of the induction culture are as follows: culturing at 27 deg.C, humidity 82% and pH 6.0 in dark for 5 days, and culturing in light at illumination intensity of 2000lux for 14 hr per day;
(2) and (3) proliferation culture: inoculating the stem section obtained by the post-treatment in the step (1) to a proliferation culture medium for culturing for 40 days, and transferring the new bud to a new proliferation culture medium for culturing, wherein the proliferation multiple reaches 4 times; wherein, the components of the proliferation culture medium are as follows: MS +1.0mg/L KT +0.2 mg/L6-BA +0.5mg/L NAA +3.5g/L agar +25g/L sucrose; the proliferation culture is carried out under the conditions that the temperature is 30 ℃, the humidity is 75% and the pH value is 6.0, the illumination intensity of the proliferation culture is 2200Lux, and the illumination time is 15 h/day;
(3) rooting culture: cutting off redundant lateral buds or adventitious buds of the cluster buds obtained by the enrichment culture in the step (2), cutting off small branches with single buds according to the height of 3cm, putting the cut small branches into a rooting culture medium for rooting culture, and taking out of a bottle after the culture for 25 days and hardening seedlings; wherein, the components of the rooting culture medium are as follows: 1/2 modified MS +1mgLIBA +2.0mg/L rooting powder +4.5g/L agar +30g/L white sugar; the rooting culture conditions are as follows: the illumination intensity is 2500lx, the illumination time is 13 hours/d, and the temperature is 30 ℃; the modified MS comprises 45 parts by weight of macronutrient elements, 0.6 parts by weight of micronutrient elements and 10 parts by weight of organic reagent; wherein, the macronutrient elements have the following components and corresponding concentrations: 200mg/L of compound sodium nitrophenolate, 210mg/L of ammonium sulfate, 390mg/L of magnesium sulfate heptahydrate and 80mg/L of brassinosteroid; the micronutrient elements and their corresponding concentrations are: 18mg/L of manganese sulfate, 4mg/L of boric acid, 2mg/L of potassium iodide, 0.4mg/L of sodium molybdate, 0.08mg/L of cobalt chloride and 0.03mg/L of copper sulfate pentahydrate; the components of the organic reagent and their corresponding concentrations are: 3mg/L of glycine, 3mg/L of nicotinic acid, 2mg/L of pyridoxine hydrochloride and 112mg/L of inositol;
(III) greenhouse seedling hardening: taking the plantlets out of the culture vessel by using tweezers, cleaning culture media at the roots of the plantlets by using tap water, planting the plantlets in a culture soil seedling tray, spraying root fixing water, spraying chlorothalonil with 1000 times of liquid after the leaf surfaces are dry, spraying 30 times of biogas liquid after 15 days, culturing in a greenhouse for 5 months, and then transplanting;
and (IV) transplanting rooted seedlings: hardening the seedlings for 30 days until the root system is changed from white to grey, transplanting the rooted seedlings, putting the rooted seedlings into clean water for rinsing during transplanting, cleaning the culture medium of the root, then soaking the rooted seedlings in a rooting agent for 10 minutes, and then transplanting the rooted seedlings into a mixed matrix container for culturing, wherein the mixed matrix is prepared by mixing the following components in a volume ratio of 1: 2: 3 vermiculite powder, weathered coal and rice bran.
Effect verification
In order to further illustrate the application value of the invention, the implemented person selects the bamboo shoot top and the axillary bud of the tortoise back with no diseases, no defects and similar sizes, and the selected bamboo shoot top and the axillary bud are divided into the following four groups for comparison test:
a first group: the tissue culture method of example 1;
second group: the tissue culture method of example 2;
third group: the tissue culture method of example 3;
and a fourth group: and (4) performing a conventional culture method.
S1: observing the tissue culture seedling conditions of the four groups in the seed culture stage, and recording the rooting rate, the rooting number, the bulb size, the leaf length, the leaf width and the comprehensive state of the bottle outlet of each group, wherein the results are shown in table 1;
TABLE 1 Table of tissue culture seedlings of four groups of Hemsleya amabilis
S2: observing the transplanting and potting stage, counting the transplanting survival rate after two months of transplanting and potting, and recording the result as shown in table 2:
table 2 growth condition table of four groups of transplanted seedlings of monstera deliciosa
The two tables show that: the monstera deliciosa cultivated by the cultivation method provided by the invention not only can obtain high-quality seedlings in a tissue culture stage, but also has good growth vigor in a field transplanting stage, and the steps are closely combined and interact with each other, so that the survival rate of the monstera deliciosa is improved, and the cultivation method is worthy of popularization.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (4)
1. A method for culturing monstera deliciosa tissue is characterized by comprising the following steps:
firstly, pretreatment of explants: cutting the stem top or axillary buds of monstera deliciosa as an explant, cutting the explant into strips of 4-6cm, soaking the explant in 70% alcohol for 3-5s, soaking in 1-1.5% sodium hypochlorite solution for 15-30min, continuously oscillating in the soaking process, washing the explant with sterile water for 4-6 times after soaking, then placing the explant on a sterile inoculation tray to blow dry the surface moisture of the explant, cutting off leaves and old cuts, cutting into 2-2.5cm stem sections with 2-3 buds, placing the stem sections with buds under tap water to wash for 30min, and sucking the stem sections with buds with filter paper for later use;
(II) culture of the culture medium:
(1) and (3) induction culture: inoculating the stem section with the bud obtained in the step on an induction culture medium, culturing for 5-10 days under the dark condition, then alternately performing induction culture according to the ratio of light to dark being 2:1, cutting off the leaf of the bud after the bud grows to 4-5cm, and cutting into stem sections of 2-3 cm;
the induction culture medium contains MS +1.5mg/L KT +0.5 mg/L6-BA +0.1mg/L IBA +4.5g/L agar +30g/L sucrose;
(2) and (3) proliferation culture: inoculating the stem section obtained by the post-treatment in the step (1) to a proliferation culture medium for culturing for 35-40 days, and transferring the new bud to a new proliferation culture medium for culturing, wherein the proliferation multiple reaches 3-4 times;
the proliferation culture medium comprises the following components: MS +1.0mg/L KT +0.2 mg/L6-BA +0.5mg/L NAA +3.5g/L agar +25g/L sucrose;
(3) rooting culture: removing redundant lateral buds or adventitious buds of the cluster buds obtained by the enrichment culture in the step (2), cutting off small branches with single buds according to the height of 1.5-3cm, putting the cut small branches into a rooting culture medium for rooting culture, and taking out of a bottle after culturing for 20-25 days to obtain seedlings;
the rooting medium comprises the following components: 1/2 modified MS +0.1-1 mg/L IBA +1.5-2.0mg/L rooting powder +4-4.5g/L agar +25-30g/L white sugar;
(III) greenhouse seedling hardening: taking the plantlet out of the culture vessel by using tweezers, cleaning a culture medium at the root of the plantlet by using tap water, then planting the plantlet into a culture soil seedling tray, spraying rooting water, spraying chlorothalonil with 800 times of liquid after the leaves are dry, spraying 20-30 times of biogas liquid after 7-15 days, and transplanting after culturing for 3-5 months in a greenhouse;
the tissue culture method further comprises the step of transplanting the rooted seedlings, and specifically comprises the following steps: hardening the seedlings for 20-30 days until the root system is changed from white to grey, transplanting the rooted seedlings, putting the rooted seedlings into clean water for rinsing during transplanting, cleaning the culture medium of the root, then soaking the rooted seedlings in a rooting agent for 5-10 minutes, and then transplanting the rooted seedlings into a container filled with a mixed matrix for culturing, wherein the mixed matrix is prepared by mixing the following components in a volume ratio of 1: 1-2: 2-3 of vermiculite powder, weathered coal and rice bran;
the improved MS in the step (3) comprises 30-45 parts by weight of macronutrient elements, 0.3-0.6 part by weight of micronutrient elements and 5-10 parts by weight of organic reagent; the macronutrient components and their corresponding concentrations are: 150mg/L of compound sodium nitrophenolate, 160mg/L of ammonium sulfate, 390mg/L of magnesium sulfate heptahydrate and 30-80mg/L of brassinosteroids; the micronutrient elements and their corresponding concentrations are: 15-18mg/L of manganese sulfate, 2-4mg/L of boric acid, 1-2mg/L of potassium iodide, 0.2-0.4mg/L of sodium molybdate, 0.05-0.08mg/L of cobalt chloride and 0.02-0.03mg/L of copper sulfate pentahydrate; the components of the organic reagent and their corresponding concentrations are: 1-3mg/L of glycine, 1-3mg/L of nicotinic acid, 1-2mg/L of pyridoxine hydrochloride and 112mg/L of inositol 102-.
2. The method for tissue culture of monstera deliciosa as claimed in claim 1, wherein the conditions for inducing culture in step (1) are as follows: the culture temperature is 23-27 deg.C, humidity is 68-82%, and pH is 5.6-6.0, dark culture is performed for 4-5 days, and light culture is performed, wherein the light intensity of light culture is 1500-.
3. The method for tissue culture of monstera deliciosa as claimed in claim 1, wherein the proliferation culture in step (2) is performed at 25-30 deg.C, humidity 50-75%, and pH 5.6-6.0, and the proliferation culture is performed with illumination intensity of 1800-2200Lux and illumination time of 10-15 h/day.
4. The method for tissue culture of monstera deliciosa as claimed in claim 1, wherein the rooting culture conditions in step (3) are as follows: the illumination intensity is 2000-.
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