CN110583486A - Method for cultivating high-stress-resistance broussonetia papyrifera - Google Patents
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
- A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
- A01G24/15—Calcined rock, e.g. perlite, vermiculite or clay aggregates
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
- A01G24/28—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing peat, moss or sphagnum
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The application discloses a cultivation method of a high-stress-resistance broussonetia papyrifera in the technical field of seedling raising of the broussonetia papyrifera, which comprises the following steps: the method comprises the following steps: carrying out partial twig cuttage and root tiller propagation on the broussonetia papyrifera to obtain a first variant progeny plant, and taking the twig of the variant progeny plant as an explant; step two: sterilizing the surface of the obtained explant; step three: carrying out primary culture on the explant; step four: carrying out rooting culture on the buds; step five: hardening the seedlings for 3d to 7d, and then transplanting the seedlings into a plug tray containing a growth culture medium; step six: planting the seedlings in the land; step seven: and performing topdressing, irrigation and insect pest prevention treatment. The method selects the mutant progeny plants bred from the yellow leaves as explants which have the high-quality genes and the mutant genes of the paper mulberry, then the sterilization is carried out to improve the stress resistance of the explants, and the paper mulberry with high stress resistance is rapidly cultivated through primary culture and rooting culture.
Description
Technical Field
The invention belongs to the technical field of paper mulberry seedling culture, and particularly relates to a cultivation method of a paper mulberry with high stress resistance.
Background
Paper mulberry, namely papermulberry, is tall in leaf and arbor, 10-20m high, dark gray in bark, dense in small branches and soft hair. The tree crown is open, the tree crown is oval to wide oval, the tree bark is smooth, light gray or grey brown, the tree is not easy to crack, the whole tree contains latex, the tree is a strong positive tree species, and the tree has strong adaptability and strong stress resistance. Although the paper mulberry is relatively rough in appearance, the paper mulberry has the advantages of dense branches and leaves, resistance, fast growth, easiness in propagation and the like, the fruits are sour and sweet, and edible, so that the paper mulberry is an important tree species for urban and rural greening, is particularly suitable for greening mining areas and barren hills, can be selected as shade trees and protective forests, is a tree species with strong toxic gas resistance, and can be planted in areas with serious air pollution.
A new feed variety hybrid paper mulberry rich in protein is cultivated in a plant key laboratory of Chinese academy of sciences by combining hybrid breeding with modern biotechnology, and is also called as white mulberry because the tree has the characteristics of high protein content and amino acid content, fast growth, high yield, multiple resistance, cutting resistance, no diseases and insect pests and the like. Because the ecological fertilizer has important economic and ecological values in the aspects of feed, papermaking, ecological greening and the like, the ecological fertilizer is planted in more than 20 provinces in China at present, so that the ecological fertilizer can help farmers to get rid of poverty and become rich and can also improve the ecological environment of poverty areas.
Through observation of the hybrid broussonetia papyrifera, it was found that very few leaves of the broussonetia papyrifera were yellow, and the growth vigor of the broussonetia papyrifera was better than that of other green-leaf broussonetia papyrifera through long-term observation. Therefore, the broussonetia papyrifera is bred to breed a broussonetia papyrifera variety with high stress resistance.
Disclosure of Invention
The invention aims to provide a cultivation method of a high stress resistance paper mulberry tree, so as to provide a high stress resistance paper mulberry tree variety.
In order to solve the above problems, the present invention provides the following solutions: a cultivation method of a high stress resistance broussonetia papyrifera includes the following steps:
the method comprises the following steps: carrying out partial twig cuttage and root shoot propagation on the broussonetia papyrifera, selecting branches with yellow single new leaves for twig cuttage to obtain a first variant progeny plant, and taking the twigs of the variant progeny plant as explants;
step two: sterilizing the surface of the obtained explant;
step three: cutting the explant into small-diameter sections with the length of 0.5-1cm and an axillary bud, and culturing on a sterilized primary culture medium until the explant grows a young bud;
step four: placing the buds on a rooting culture medium for rooting culture, gradually growing white radicles after culturing for 2-3 weeks, and hardening seedlings when 6-8 roots grow out;
step five: hardening seedlings for 3-7 days, transplanting the seedlings into a plug tray containing a growth culture medium, wherein the growth culture medium consists of vermiculite, peat and perlite, placing the plug tray in a greenhouse, and carrying out light, temperature, water and fertilizer management on the seedlings until the height of the seedlings is 10-15 cm;
step six: planting the seedlings in a cultivation land with the row spacing of 3-4m multiplied by the plant spacing of 1-1.5 m;
step seven: topdressing, irrigation and pest control treatment.
The principle and the beneficial effects of the scheme are as follows: the method selects the mutant progeny plants bred from the yellow leaves as explants which have the high-quality genes and the mutant genes of the paper mulberry, then the sterilization is carried out to improve the stress resistance of the explants, and the paper mulberry with high stress resistance is rapidly cultivated through primary culture and rooting culture.
Further, the primary culture medium is prepared by adding 2, 4-dichlorophenoxyacetic acid, cytokinin, 6-benzylaminopurine and 1-naphthylacetic acid into an MS culture medium, wherein the concentration of the 2, 4-dichlorophenoxyacetic acid is 1.8-2.3mg/L, the concentration of the cytokinin is 0.8-1.3mg/L, the concentration of the 6-benzylaminopurine is 0.9-1.2mg/L, and the concentration of the 1-naphthylacetic acid is 0.3-0.6 mg/L. The primary culture medium of the scheme is adopted to culture the explant, so that the formation of the buds can be effectively promoted.
Furthermore, the rooting medium is MS medium added with indolebutyric acid, spermidine, sucrose and agar, wherein the concentration of indolebutyric acid is 0.1-1.0mg/L, the concentration of spermidine is 5.0-20.0mg/L, the concentration of sucrose is 15-25g/L, and the concentration of agar is 6.5-8.0 g/L. The rooting culture medium can rapidly promote the generation of seedling roots, the seedling rooting rate is 100%, and 20 balance rooting trees are 5.5 roots.
Further, the culture condition of the rooting culture is that the illumination time is 10-12h/d, and the temperature is 25 +/-3 ℃.
Further, the seedling exercising treatment comprises the following steps: placing the rooted tissue culture seedling of the hybrid paper mulberry together with a tissue culture bottle in the sun, loosening the bottle cap after 4-5 days, taking off the bottle cap 3 days after the cap is loosened to enable the tissue culture bottle seedling to be in a fully open state with the external environment, and transplanting after 2 days; taking out tissue-cultured broussonetia papyrifera seedlings from a culture medium, cleaning the culture medium, soaking the tissue-cultured broussonetia papyrifera seedlings in 1000-fold diluted carbendazim or chlorothalonil solution for 10-20 minutes, then inoculating the hybrid broussonetia papyrifera seedlings into a matrix which is sterilized by three thousand of formaldehyde and is mixed with garden soil, vermiculite and a small amount of perlite for culturing, wherein the ratio of the vermiculite to the garden soil to the perlite is 6:3:1, the seedling hardening condition is that the temperature is 25 ℃ and the humidity is 75-90%, covering a plastic film on the upper layer of the plants for water retention, ventilating and taking down the plastic film gradually after culturing for 10 days.
Detailed Description
The following is further detailed by the specific embodiments:
example 1: a cultivation method of a high stress resistance broussonetia papyrifera includes the following steps:
the method comprises the following steps: carrying out partial twig cuttage and root shoot propagation on the broussonetia papyrifera, selecting branches with yellow single new leaves for twig cuttage to obtain a first variant progeny plant, and taking the twigs of the variant progeny plant as explants;
step two: sterilizing the surface of the obtained explant;
step three: cutting the explant into small diameter sections with the length of 0.5cm and an axillary bud, and culturing on a sterilized primary culture medium until the explant grows out a young bud, wherein the primary culture medium is prepared by adding 2, 4-dichlorophenoxyacetic acid, cytokinin, 6-benzylaminopurine and 1-naphthylacetic acid into an MS culture medium, the concentration of the 2, 4-dichlorophenoxyacetic acid is 1.8mg/L, the concentration of the cytokinin is 0.8mg/L, the concentration of the 6-benzylaminopurine is 0.9mg/L, and the concentration of the 1-naphthylacetic acid is 0.3 mg/L. (ii) a
Step four: and (3) placing the buds on a rooting culture medium for rooting culture under the conditions that the illumination time is 10h/d and the temperature is 22 ℃. The rooting culture medium is an MS culture medium, indolebutyric acid, spermidine, sucrose and agar are added, wherein the concentration of indolebutyric acid is 0.1mg/L, the concentration of spermidine is 5.0mg/L, the concentration of sucrose is 15g/L, the concentration of agar is 6.5g/L, white young roots grow gradually after 2-3 weeks of culture, and the seedling hardening treatment is carried out when 6 roots grow out, and comprises the following steps: placing the rooted tissue culture seedling of the hybrid paper mulberry together with a tissue culture bottle in the sun, loosening the bottle cap after 4 days, taking off the bottle cap 3 days after the cap is loosened, enabling the tissue culture bottle seedling to be in a fully open state with the external environment, and transplanting after 2 days; taking out tissue-cultured broussonetia papyrifera seedlings from a culture medium, cleaning the culture medium, soaking the tissue-cultured broussonetia papyrifera seedlings in 1000-fold diluted carbendazim or chlorothalonil solution for 10 minutes, then inoculating the hybrid broussonetia papyrifera seedlings into a matrix which is sterilized by trioxymethylene and is mixed with garden soil, vermiculite and a small amount of perlite for culturing, wherein the ratio of the vermiculite to the garden soil to the perlite is 6:3:1, the seedling hardening condition is that the temperature is 25 ℃ and the humidity is 75%, covering a plastic film on the upper layer of the plants for water retention, ventilating and ventilating by combining the local environment, and gradually taking down the plastic film after culturing for 10 days;
step five: hardening 3dd seedlings, transplanting the seedlings into a plug tray containing a growth culture medium, wherein the growth culture medium consists of vermiculite, peat and perlite, placing the plug tray in a greenhouse, and carrying out light, temperature, water and fertilizer management on the seedlings until the height of the seedlings is 10 cmcm;
step six: planting the seedlings in a cultivation land with the row spacing of 3m multiplied by the plant spacing of 1 m;
step seven: topdressing, irrigation and pest control treatment.
Example 2: a cultivation method of a high stress resistance broussonetia papyrifera includes the following steps:
the method comprises the following steps: carrying out partial twig cuttage and root shoot propagation on the broussonetia papyrifera, selecting branches with yellow single new leaves for twig cuttage to obtain a first variant progeny plant, and taking the twigs of the variant progeny plant as explants;
step two: sterilizing the surface of the obtained explant;
step three: cutting the explant into small diameter sections with the length of 0.8cm and an axillary bud, and culturing on a sterilized primary culture medium until the explant grows out a young bud, wherein the primary culture medium is prepared by adding 2, 4-dichlorophenoxyacetic acid, cytokinin, 6-benzylaminopurine and 1-naphthylacetic acid into an MS culture medium, the concentration of the 2, 4-dichlorophenoxyacetic acid is 2.0mg/L, the concentration of the cytokinin is 1.1mg/L, the concentration of the 6-benzylaminopurine is 1.05mg/L, and the concentration of the 1-naphthylacetic acid is 0.45 mg/L. (ii) a
Step four: and (3) placing the buds on a rooting culture medium for rooting culture under the conditions that the illumination time is 11h/d and the temperature is 25 ℃. The rooting culture medium is an MS culture medium, indolebutyric acid, spermidine, sucrose and agar are added, wherein the concentration of indolebutyric acid is 0.5mg/L, the concentration of spermidine is 13mg/L, the concentration of sucrose is 20g/L, the concentration of agar is 7.2g/L, white young roots grow gradually after 2-3 weeks of culture, and the hardening treatment is carried out when 7 roots grow out, and the hardening treatment is as follows: placing the rooted tissue culture seedling of the hybrid paper mulberry together with a tissue culture bottle in the sun, loosening the bottle cap after 4 days, taking off the bottle cap 3 days after the cap is loosened, enabling the tissue culture bottle seedling to be in a fully open state with the external environment, and transplanting after 2 days; taking out tissue-cultured broussonetia papyrifera seedlings from a culture medium, cleaning the culture medium, soaking the tissue-cultured broussonetia papyrifera seedlings in 1000-fold diluted carbendazim or chlorothalonil solution for 15 minutes, inoculating the hybrid broussonetia papyrifera seedlings into a matrix which is sterilized by trioxymethylene and is mixed with garden soil, vermiculite and a small amount of perlite for culturing, wherein the ratio of the vermiculite to the garden soil to the perlite is 6:3:1, the seedling hardening condition is that the temperature is 25 ℃ and the humidity is 82%, covering a plastic film on the upper layer of the plants for water retention, ventilating and ventilating by combining the local environment, and gradually taking down the plastic film after culturing for 10 days. (ii) a
Step five: hardening the seedlings for 5 days, transplanting the seedlings into a plug tray containing a growth culture medium, wherein the growth culture medium consists of vermiculite, peat and perlite, placing the plug tray in a greenhouse, and carrying out light, temperature, water and fertilizer management on the seedlings until the height of the seedlings is 13 cm;
step six: planting the seedlings in a cultivation land with the row spacing of 3m multiplied by the plant spacing of 1.5 m;
step seven: topdressing, irrigation and pest control treatment.
Example 3: a cultivation method of a high stress resistance broussonetia papyrifera includes the following steps:
the method comprises the following steps: carrying out partial twig cuttage and root shoot propagation on the broussonetia papyrifera, selecting branches with yellow single new leaves for twig cuttage to obtain a first variant progeny plant, and taking the twigs of the variant progeny plant as explants;
step two: sterilizing the surface of the obtained explant;
step three: cutting an explant into small-diameter sections with the length of 1cm and an axillary bud, and culturing the small-diameter sections on a sterilized primary culture medium until the explant grows to form a young bud, wherein the primary culture medium is prepared by adding 2, 4-dichlorophenoxyacetic acid, cytokinin, 6-benzylaminopurine and 1-naphthylacetic acid into an MS culture medium, the concentration of the 2, 4-dichlorophenoxyacetic acid is 2.3mg/L, the concentration of the cytokinin is 1.3mg/L, the concentration of the 6-benzylaminopurine is 1.2mg/L, and the concentration of the 1-naphthylacetic acid is 0.6 mg/L;
step four: and placing the buds on a rooting culture medium for rooting culture under the conditions that the illumination time is 12h/d and the temperature is 28 ℃. The rooting culture medium is an MS culture medium, indolebutyric acid, spermidine, sucrose and agar are added, wherein the concentration of indolebutyric acid is 1.0mg/L, the concentration of spermidine is 20.0mg/L, the concentration of sucrose is 25g/L, the concentration of agar is 8.0g/L, white young roots grow gradually after 2-3 weeks of culture, and the seedling hardening treatment is carried out when 8 roots grow out, and comprises the following steps: placing the rooted tissue culture seedling of the hybrid paper mulberry together with a tissue culture bottle in the sun, loosening the bottle cap after 5 days, taking off the bottle cap 3 days after the cap is loosened, enabling the tissue culture bottle seedling to be in a fully open state with the external environment, and transplanting after 2 days; taking out tissue-cultured broussonetia papyrifera seedlings from a culture medium, cleaning the culture medium, soaking the tissue-cultured broussonetia papyrifera seedlings in 1000-fold diluted carbendazim or chlorothalonil solution for 20 minutes, inoculating the hybrid broussonetia papyrifera seedlings into a matrix which is sterilized by trioxymethylene and is mixed with garden soil, vermiculite and a small amount of perlite for culturing, wherein the ratio of the vermiculite to the garden soil to the perlite is 6:3:1, the seedling hardening condition is that the temperature is 25 ℃ and the humidity is 90%, covering a plastic film on the upper layer of the plants for water retention, ventilating and ventilating by combining the local environment, and gradually taking down the plastic film after culturing for 10 days. (ii) a
Step five: hardening the seedlings for 7 days, transplanting the seedlings into a plug tray containing a growth culture medium, wherein the growth culture medium consists of vermiculite, peat and perlite, placing the plug tray in a greenhouse, carrying out light, temperature, water and fertilizer management on the seedlings, and culturing the seedlings until the height of the seedlings is 15 cm;
step six: planting the seedlings in a cultivation land with the row spacing of 3m multiplied by the plant spacing of 1.5 m;
step seven: topdressing, irrigation and pest control treatment.
Comparing the tissue culture seedlings of the broussonetia papyrifera produced in the examples 1-3 with the tissue culture seedlings of the broussonetia papyrifera cultivated by the traditional method after planting, the data are as follows:
example 1: in 500 broussonetia papyrifera planted in each mu of land, the average number of dead broussonetia papyrifera is 16, and the survival rate of the broussonetia papyrifera is 96.8%.
Example 2: in 500 broussonetia papyrifera planted in each mu of land, the average dead broussonetia papyrifera is 10, and the survival rate of the broussonetia papyrifera is 98%.
Example 3: the average number of dead broussonetia papyrifera among 500 broussonetia papyrifera planted in each mu of land is 17, and the survival rate of the broussonetia papyrifera is 96.6%.
In the case of no nutrient solution injection on the broussonetia papyrifera, the average dead broussonetia papyrifera among 500 broussonetia papyrifera planted in each mu of land is 41, and the survival rate of the broussonetia papyrifera is 91.8%.
The data clearly show that the survival rate of the broussonetia papyrifera cultivated by the method is far higher than that of broussonetia papyrifera seedlings cultivated by traditional seedling cultivation, and the method proves that the broussonetia papyrifera has high stress resistance and strong adaptability to new environments.
Claims (5)
1. A cultivation method of a high stress resistance broussonetia papyrifera is characterized by comprising the following steps: the method comprises the following steps:
the method comprises the following steps: carrying out partial twig cuttage and root shoot propagation on the broussonetia papyrifera, selecting branches with yellow single new leaves for twig cuttage to obtain a first variant progeny plant, and taking the twigs of the variant progeny plant as explants;
step two: sterilizing the surface of the obtained explant;
step three: cutting the explant into small-diameter sections with the length of 0.5-1cm and an axillary bud, and culturing on a sterilized primary culture medium until the explant grows a young bud;
step four: placing the buds on a rooting culture medium for rooting culture, gradually growing white radicles after culturing for 2-3 weeks, and hardening seedlings when 6-8 roots grow out;
step five: hardening seedlings for 3-7 days, transplanting the seedlings into a plug tray containing a growth culture medium, wherein the growth culture medium consists of vermiculite, peat and perlite, placing the plug tray in a greenhouse, and carrying out light, temperature, water and fertilizer management on the seedlings until the height of the seedlings is 10-15 cm;
step six: planting the seedlings in a cultivation land with the row spacing of 3-4m multiplied by the plant spacing of 1-1.5 m;
step seven: topdressing, irrigation and pest control treatment.
2. The method for cultivating a high stress resistance broussonetia papyrifera according to claim 1, wherein the method comprises the following steps: the primary culture medium is prepared by adding 2, 4-dichlorophenoxyacetic acid, cytokinin, 6-benzylamino adenine and 1-naphthylacetic acid into an MS culture medium, wherein the concentration of the 2, 4-dichlorophenoxyacetic acid is 1.8-2.3mg/L, the concentration of the cytokinin is 0.8-1.3mg/L, the concentration of the 6-benzylamino adenine is 0.9-1.2mg/L, and the concentration of the 1-naphthylacetic acid is 0.3-0.6 mg/L.
3. The method for cultivating the high stress resistance broussonetia papyrifera according to claim 2, wherein the method comprises the following steps: the rooting culture medium is an MS culture medium added with indolebutyric acid, spermidine, sucrose and agar, wherein the concentration of the indolebutyric acid is 0.1-1.0mg/L, the concentration of the spermidine is 5.0-20.0mg/L, the concentration of the sucrose is 15-25g/L, and the concentration of the agar is 6.5-8.0 g/L.
4. The method for cultivating a high stress resistance broussonetia papyrifera according to claim 3, wherein: the culture conditions of the rooting culture are that the illumination time is 10-12h/d and the temperature is 25 +/-3 ℃.
5. The method for cultivating a high stress resistance broussonetia papyrifera according to claim 4, wherein: the seedling exercising treatment comprises the following steps: placing the rooted tissue culture seedling of the hybrid paper mulberry together with a tissue culture bottle in the sun, loosening the bottle cap after 4-5 days, taking off the bottle cap 3 days after the cap is loosened to enable the tissue culture bottle seedling to be in a fully open state with the external environment, and transplanting after 2 days; taking out tissue-cultured broussonetia papyrifera seedlings from a culture medium, cleaning the culture medium, soaking the tissue-cultured broussonetia papyrifera seedlings in 1000-fold diluted carbendazim or chlorothalonil solution for 10-20 minutes, then inoculating the hybrid broussonetia papyrifera seedlings into a matrix which is sterilized by three thousand of formaldehyde and is mixed with garden soil, vermiculite and a small amount of perlite for culturing, wherein the ratio of the vermiculite to the garden soil to the perlite is 6:3:1, the seedling hardening condition is that the temperature is 25 ℃ and the humidity is 75-90%, covering a plastic film on the upper layer of the plants for water retention, ventilating and taking down the plastic film gradually after culturing for 10 days.
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Application publication date: 20191220 |