CN108812260B - Tissue culture and rapid seedling method for paper mulberry root hairs - Google Patents
Tissue culture and rapid seedling method for paper mulberry root hairs Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
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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
- A01G2/00—Vegetative propagation
- A01G2/10—Vegetative propagation by means of cuttings
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures
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Abstract
The invention relates to the field of agriculture, and discloses a tissue culture and rapid seedling method for broussonetia papyrifera root hairs, which comprises the following steps: 1) cutting root hairs: digging up soil of a female broussonetia papyrifera, cleaning root hairs of the female broussonetia papyrifera, cutting the root hairs with the diameter of 1-5mm into 3-4cm long, and soaking the root hairs in a growth promoting agent; 2) preparing a soilless nutrient substrate; 3) tissue culture; 4) hardening seedlings; 5) transplanting in a field: after hardening seedlings for 28-32 days, transplanting the root hairs to a field. The method adopts the broussonetia papyrifera root hairs for tissue culture, has short seedling culture period (only about 45 days, the growth of the broussonetia papyrifera root hairs is more than 30cmn, the broussonetia papyrifera root hairs can be transplanted in a field), has low cost and high survival rate, and ensures that the varieties are not easy to degenerate in the later period.
Description
Technical Field
The invention relates to the field of agriculture, in particular to a tissue culture and rapid seedling method for paper mulberry root hairs.
Background
The paper mulberry is a common tree species which inhabits in every noon corner in China, but at present, the paper mulberry becomes a high-protein forest which stores grains in forests and stores grains in skills through modern biotechnology, has profound significance on national grain safety, can make the best use of things, benefits human beings and benefits society.
The paper mulberry has strong adaptability, fast growth, high yield, strong vitality, developed root system and simple and extensive management, and the planted paper mulberry becomes a forest, and has important functions in aspects of difficult afforestation (saline-alkali soil, desert, stony desert and desert regions), development of understory economy, recovery of mine vegetation, purification and treatment of atmospheric pollution and the like. The broussonetia papyrifera has stress resistance, is concentrated to show that lateral roots are developed and are concentrated on the earth surface by more than 30cm, has extremely strong capabilities of resisting surface runoff, controlling water and soil loss and preventing soil stony desertification and desertification, and is drought-resistant, saline-alkali resistant and barren resistant. Therefore, the method becomes the first choice tree species for afforestation, wind prevention and sand fixation and water and soil loss treatment of barren mountains, barren slopes, barren lands, barren beaches and saline-alkali lands.
The application field of the paper mulberry is wide: tender branches and leaves of the broussonetia papyrifera can be processed into biological feed by a biotechnology, and the biological feed is used as high-protein high-quality feed for livestock and poultry; the bark is high-grade natural bast fiber, is a raw material of high-grade paper pulp and textile fiber, and has high price and large market demand; the wood rod can be used as the raw material of common paper pulp and artificial boards; the tree bud and the tree root can be processed into health food, health care products and medicines. The paper mulberry also has the capability of rapid growth, is simple to manage, can be harvested in the same year, sprouts new branches after cutting, can be harvested 3-5 times in one year, and can achieve the sustainable income effects of one-year cultivation and more than twenty-year harvest. Through the steps of paper mulberry planting, biotechnology processing, livestock and poultry breeding, livestock and poultry manure waste fermentation, the ecological cycle and beautiful country construction of paper mulberry forests are achieved, and ecological, economic and social benefits are integrated. As a woody feed, the broussonetia papyrifera has large biomass, is resistant to cutting, can be harvested for multiple times in one year, and can be harvested for multiple years after being planted once. It has rich nutrient components, and the crude protein content of tender branches and leaves is more than 26%; is rich in calcium, selenium, zinc, ferrum and other microelements and high protein amino acid. Has strong palatability to livestock, high utilization rate of branches and leaves, various feeding forms, capability of fermenting and ensiling, and capability of being prepared into commercial feed (powder or pellet feed) for self-use or distant marketing. The livestock, poultry and aquatic product eating the broussonetia papyrifera has faint scent meat quality, no peculiar smell, good taste, greatly reduced cholesterol, high lean meat percentage and strong market competitiveness.
In conclusion, the paper mulberry is a compatible tree species integrating ecological property and economical property, and is also a preferred tree species for implementing novel planting and raising integrated forest and pasture to produce protein feed. The requirements of the saplings are vigorous,
in the prior art, tender shoots or branches are generally selected for tissue culture of paper mulberry, and the problems of high facility and equipment requirements, slow rooting, long seedling growing period, high cost, low survival rate and easy degeneration of later-stage varieties generally exist.
The Chinese patent with the application number of 201210070464.0 discloses a simple tissue culture method for broussonetia papyrifera, which comprises the steps of preparation of a bacteriostatic agent, disinfection of a culture container, preparation of a culture medium, induction culture, multiplication culture, rooting culture and bottle seedling transplantation. According to the method, the paper mulberry branches are used as tissue culture objects, at least 26 days are needed from the beginning of tissue culture to the transplanting of bottle-loaded rooting seedlings into a greenhouse, at least more than 2 months are needed for field planting, and the seedling culture period is long.
Disclosure of Invention
In order to solve the technical problems, the invention provides a tissue culture and rapid seedling method for paper mulberry root hairs, the tissue culture and rapid seedling method for paper mulberry root hairs is adopted, the seedling period is short (the seedling period is as long as about 45 days and is more than 30cmn, the seedling can be transplanted in a field), the cost is low, the survival rate is high, and the varieties in the later period are not easy to degenerate.
The specific technical scheme of the invention is as follows: a tissue culture and rapid seedling method for paper mulberry root hairs is characterized by comprising the following steps:
1) cutting root hairs: digging up soil of female broussonetia papyrifera, cleaning root hairs of the female broussonetia papyrifera, cutting the root hairs with the diameter of 1-5mm into 3-4cm, and soaking in a growth promoting agent.
2) And (4) preparation of a soilless nutrient substrate.
3) Tissue culture: inserting the soaked root hairs into a soilless nutrient substrate, and then spraying a growth promoting agent to moisten the soilless nutrient substrate; performing tissue culture in a tissue culture room at 23-27 deg.C and 65-75% humidity; spraying water and lighting during tissue culture.
4) Hardening seedlings: after tissue culture is carried out for 13-17 days, transplanting the root hairs with the length of more than 10cm into a black film hardening box with organic fertilizer nutrient soil, and hardening seedlings in a lighting greenhouse covered with a lighting plastic film; spraying water and shading during seedling hardening, and controlling the temperature in the greenhouse to be above 23-27 ℃.
5) Transplanting in a field: after hardening seedlings for 28-32 days, transplanting the root hairs with the length of more than 30cm into a field.
In the prior art, the tissue culture of the paper mulberry usually selects the tender bud or the branch, but does not select the root or the root hair, because the germination capacity of the tissue is the most important in the tissue culture, and compared with the tender bud or the branch, the germination rate of the root or the root hair is far lower. However, the group of the invention discovers that the adoption of tender shoots or branches for tissue culture also has the following defects after research: the quantity of the tender shoots on one paper mulberry is limited, and the survival rate of the female paper mulberry is low after the tender shoots or branches of the female paper mulberry are cut. Also, the rootage rate of shoots or branches is slow, which is also affected by seasons. In addition, the group of the invention also finds that the paper mulberry seedling obtained by tissue culture of tender shoots or branches has the problem that the variety is easy to degrade in the later period.
Therefore, the root hair is selected as the tissue culture object for the first time creatively, the root hair rooting rate is good, the number of roots of the paper mulberry is large, one paper mulberry can collect a large number of root hairs, the influence on the survival rate of the female paper mulberry is low after the root hairs are subtracted, the tissue culture time is not influenced by seasons, and the paper mulberry seedling obtained by selecting the root hair tissue culture is not easy to degrade in the later period. In order to solve the technical problem that the root hair germination efficiency is low, after research, the team of the invention strictly limits the size of the root hair to be 1-5mm in diameter and cuts the root hair to be 3-4cm in length, and the reason for selecting the root hair with the size is that a large amount of research discovers that the root hair in the diameter range has the best germination efficiency, the germination efficiency is reduced when the diameter is too large, and the survival rate is influenced when the diameter is too small. Then, the root hairs are soaked in the growth promoting agent specially prepared by the invention, so that the germination efficiency and rooting rate of the root hairs can be greatly improved. Finally, the tissue culture environmental conditions need to be strictly controlled during seedling hardening, and the purpose of rapid seedling culture of the root hairs can be achieved under the cooperation of the above factors.
Preferably, in the step 1), before excavating the parent broussonetia papyrifera, cutting off branches 35-45cm above the ground; and after cutting off the root hairs, soaking the rest roots in the growth promoting agent for 3-7min, immediately planting and watering again and applying organic fertilizer for daily weeding.
After the treatment, the survival rate of the maternal broussonetia papyrifera can be further improved, and seedlings can be excavated in the next year.
Preferably, in step 1), the maternal broussonetia papyrifera is an aerospace hybrid broussonetia papyrifera variety of Chinese academy of sciences.
The broussonetia papyrifera of the variety has strong growth adaptability, repeated mowing resistance, strong regeneration capability, wide and thick leaves, good taste and high protein.
Preferably, the cutting of the root hairs in step 1) is performed in a sterile environment.
Preferably, in step 1), the growth promoter is prepared by the following method: taking paper mulberry root tissues, crushing, carrying out biological enzymolysis fermentation to obtain active peptide freeze-dried powder, then preparing into a solution with the mass concentration of 15-25%, and respectively adding 0.1-0.3% of colchicine, 0.5-1.5% of indole auxin and 1-3% of naphthoic acid auxin by mass of the solution to prepare the growth promoter.
The growth promoter is specially designed for paper mulberry root hairs, can promote the root hairs to rapidly take roots, germinate and divide leaves under the aseptic condition and at the proper temperature, enables the root hairs to be used as tissue culture objects to be possible, and can greatly shorten the tissue culture period.
Preferably, in the step 1), the root hairs are soaked for 50-70 min.
Preferably, in the step 2), the soilless culture substrate comprises the following components in percentage by mass: 8-12% of rapeseed meal powder after fermentation treatment and 8-12% of tea seed meal powder after fermentation treatment; 18-22% of fermented broussonetia papyrifera hard branch powder; 55-65% of vermiculite powder; the thickness of the soilless culture substrate is 5.5-6.5 cm.
Preferably, the preparation method of the soilless culture substrate comprises the following steps: weighing the components according to the proportion, stirring, mixing, sieving, sterilizing at the temperature of 115-125 ℃, and then putting the soilless nutrient substrate into a plastic box.
Preferably, in the step 3), the root hairs are inserted into the soilless culture substrate by 1.5-2.5cm and the distance between the root hairs and the soilless culture substrate is 2-3 cm.
Preferably, in the step 4), when the external natural temperature is above-10 ℃ in winter, a plastic film small shed is arranged in the greenhouse, the small shed is covered with non-woven fabrics, and foliar fertilizer is sprayed at intervals of 8-12 days.
The seedlings do not fall off leaves and grow in spring by the treatment according to the method.
Preferably, in the step 5), before the field transplantation, the growth promoting agent is sprayed on the black membrane hardening box, so that the black membrane hardening box is thoroughly sprayed.
Compared with the prior art, the invention has the beneficial effects that: the method adopts the broussonetia papyrifera root to carry out tissue culture, has short seedling culture period (only about 45 days is longer than 30cmn, the field transplantation can be carried out), has low cost and high survival rate, and the later-stage variety is not easy to degrade and is not influenced by seasons.
Detailed Description
The present invention will be further described with reference to the following examples.
General examples
A tissue culture and rapid seedling method for paper mulberry root hairs comprises the following steps:
1) cutting root hairs: cutting off branches 35-45cm above ground of mother broussonetia papyrifera (a species of space cross broussonetia papyrifera of Chinese academy of sciences), digging up soil of the mother broussonetia papyrifera, cleaning root hairs of the mother broussonetia papyrifera, cutting the root hairs with the diameter of 1-5mm into 3-4cm in a sterile environment, and soaking in a growth promoter for 50-70 min. And after cutting off the root hairs, soaking the rest roots in the growth promoting agent for 3-7min, immediately planting and watering again and applying organic fertilizer for daily weeding.
The preparation method of the growth promoter comprises the following steps: taking paper mulberry root tissues, crushing, carrying out biological enzymolysis fermentation to obtain active peptide freeze-dried powder, then preparing into a solution with the mass concentration of 15-25%, and respectively adding 0.1-0.3% of colchicine, 0.5-1.5% of indole auxin and 1-3% of naphthoic acid auxin by mass of the solution to prepare the growth promoter.
2) Preparing a soilless nutrient substrate: the soilless culture substrate comprises the following components in percentage by mass: 8-12% of rapeseed meal powder after fermentation treatment and 8-12% of tea seed meal powder after fermentation treatment; 18-22% of fermented broussonetia papyrifera hard branch powder; 55-65% of vermiculite powder; the thickness of the soilless culture substrate is 5.5-6.5 cm. The preparation method comprises the following steps: weighing the components according to the proportion, stirring, mixing, sieving, sterilizing at the temperature of 115-125 ℃, and then putting the soilless nutrient substrate into a plastic box.
3) Tissue culture: inserting the soaked root hairs into the soilless culture substrate for 1.5-2.5cm, and keeping the distance between the root hairs and the soilless culture substrate for 2-3 cm. Then, spraying a growth promoting agent to wet the soilless nutrient substrate; performing tissue culture in a tissue culture room at 23-27 deg.C and 65-75% humidity; spraying water and lighting during tissue culture.
4) Hardening seedlings: after about 13-17 days of tissue culture, transplanting the root hairs with the length of more than 10cm into a black film hardening box with organic fertilizer nutrient soil, and hardening the seedlings in a lighting greenhouse covered with a lighting plastic film; spraying water and shading during seedling hardening, and controlling the temperature in the greenhouse to be above 23-27 ℃. Wherein, when the outside natural temperature is above-10 ℃ in winter, a plastic film small shed is arranged in the greenhouse, the small shed is covered with non-woven fabrics, and foliar fertilizer is sprayed at intervals of 8-12 days.
5) Transplanting in a field: after hardening off for about 28-32 days, transplanting the root hair with the length of more than 30cm to a field. Wherein, the growth promoting agent is sprayed on the black film hardening box before transplanting, so that the black film hardening box is thoroughly sprayed.
Example 1
A tissue culture and rapid seedling method for paper mulberry root hairs comprises the following steps:
1) cutting root hairs: cutting off branches of female broussonetia papyrifera (a species of space cross broussonetia papyrifera of Chinese academy of sciences) about 40cm above ground, digging up soil of the female broussonetia papyrifera, cleaning root hairs of the female broussonetia papyrifera, cutting the root hairs with the diameter of 1-5mm into 3-4cm in a sterile environment, and soaking in a growth promoter for 60 min. And after cutting off the root hairs, soaking the rest roots in the growth promoting agent for 5min, immediately planting and watering again and applying organic fertilizer for daily weeding.
The preparation method of the growth promoter comprises the following steps: taking paper mulberry root tissues, crushing, carrying out biological enzymolysis fermentation to obtain active peptide freeze-dried powder, then preparing into a solution with the mass concentration of 20%, and respectively adding colchicine accounting for 0.2% of the mass of the solution, indole auxin accounting for 1% of the mass of the solution and naphthoic auxin accounting for 2% of the mass of the solution to prepare the growth promoter.
2) Preparing a soilless nutrient substrate: the soilless culture substrate comprises the following components in percentage by mass: 10% of rapeseed meal powder subjected to fermentation treatment and 10% of tea seed meal powder subjected to fermentation treatment; 20% of fermentation-treated paper mulberry hard branch powder; 60% of vermiculite powder; the thickness of the soilless culture substrate is 6 cm. The preparation method comprises the following steps: weighing the components according to the proportion, stirring, mixing, sieving, sterilizing at 121 ℃ and then putting the soilless nutrient substrate into a plastic box.
3) Tissue culture: and inserting the soaked root hairs into the soilless culture substrate for 2cm, wherein the distance between the root hairs and the soilless culture substrate is 2.5 cm. Then, spraying a growth promoting agent to wet the soilless nutrient substrate; carrying out tissue culture in a tissue culture room at the temperature of 25 ℃ and the humidity of 70%; spraying water and lighting during tissue culture.
4) Hardening seedlings: after 15 days or so, transplanting the root hairs with the length of more than 10cm into a black film hardening box with organic fertilizer nutrient soil, and hardening the seedlings in a lighting greenhouse covered with a lighting plastic film; spraying water and shading during seedling hardening, and controlling the temperature in the greenhouse to be more than 25 ℃.
5) Transplanting in a field: after hardening off for about 30 days, transplanting the root hair with the length of more than 30cm to a field. Wherein, the growth promoting agent is sprayed on the black film hardening box before transplanting, so that the black film hardening box is thoroughly sprayed.
Example 2
A tissue culture and rapid seedling method for paper mulberry root hairs comprises the following steps:
1) cutting root hairs: cutting off branches 35cm above ground of mother broussonetia papyrifera (a species of space cross broussonetia papyrifera of Chinese academy of sciences), digging up soil of the mother broussonetia papyrifera, cleaning root hairs of the mother broussonetia papyrifera, cutting the root hairs with the diameter of 1-5mm into 3-4cm in a sterile environment, and soaking in a growth promoter for 50 min. And after cutting off the root hairs, soaking the rest roots in the growth promoting agent for 3min, immediately planting and watering again and applying organic fertilizer for daily weeding.
The preparation method of the growth promoter comprises the following steps: taking paper mulberry root tissues, crushing, carrying out biological enzymolysis fermentation to obtain active peptide freeze-dried powder, then preparing into a solution with the mass concentration of 15%, and respectively adding colchicine accounting for 0.1% of the mass of the solution, indole auxin accounting for 0.5% of the mass of the solution and naphthoic auxin accounting for 1% of the mass of the solution to prepare the growth promoter.
2) Preparing a soilless nutrient substrate: the soilless culture substrate comprises the following components in percentage by mass: 8% of rapeseed meal powder subjected to fermentation treatment and 12% of tea seed meal powder subjected to fermentation treatment; 18% of fermented broussonetia papyrifera hard branch powder; 62% of vermiculite powder; the thickness of the soilless culture substrate is 5.5 cm. The preparation method comprises the following steps: weighing the components according to the proportion, stirring, mixing, sieving, sterilizing at 121 ℃ and then putting the soilless nutrient substrate into a plastic box.
3) Tissue culture: inserting the soaked root hairs into the soilless culture substrate for 1.5cm, and keeping the distance between the root hairs and the soilless culture substrate for 2 cm. Then, spraying a growth promoting agent to wet the soilless nutrient substrate; carrying out tissue culture in a tissue culture room at 23 ℃ and 65% humidity; spraying water and lighting during tissue culture.
4) Hardening seedlings: after 15 days or so, transplanting the root hairs with the length of more than 10cm into a black film hardening box with organic fertilizer nutrient soil, and hardening the seedlings in a lighting greenhouse covered with a lighting plastic film; spraying water and shading during seedling hardening, and controlling the temperature in the greenhouse to be more than 23 ℃.
5) Transplanting in a field: after hardening off for about 30 days, transplanting the root hair with the length of more than 30cm to a field. Wherein, the growth promoting agent is sprayed on the black film hardening box before transplanting, so that the black film hardening box is thoroughly sprayed.
Example 3
A tissue culture and rapid seedling method for paper mulberry root hairs comprises the following steps:
1) cutting root hairs: cutting off the branches 45cm above the ground of the female broussonetia papyrifera (a space cross broussonetia papyrifera variety of Chinese academy of sciences), digging up soil of the female broussonetia papyrifera, cleaning root hairs of the female broussonetia papyrifera, cutting the root hairs with the diameter of 1-5mm into 3-4cm in a sterile environment, and soaking in a growth promoter for 70 min. And after cutting off the root hairs, soaking the rest roots in the growth promoting agent for 7min, immediately planting and watering again and applying organic fertilizer for daily weeding.
The preparation method of the growth promoter comprises the following steps: taking paper mulberry root tissues, crushing, carrying out biological enzymolysis fermentation to obtain active peptide freeze-dried powder, then preparing into a solution with the mass concentration of 25%, and respectively adding colchicine accounting for 0.3% of the mass of the solution, indole auxin accounting for 1.5% of the mass of the solution and naphthoic auxin accounting for 3% of the mass of the solution to prepare the growth promoter.
2) Preparing a soilless nutrient substrate: the soilless culture substrate comprises the following components in percentage by mass: 12% of rapeseed meal powder subjected to fermentation treatment and 8% of tea seed meal powder subjected to fermentation treatment; 22% of fermented paper mulberry hard branch powder; 58% of vermiculite powder; the thickness of the soilless culture substrate is 6.5 cm. The preparation method comprises the following steps: weighing the components according to the proportion, stirring, mixing, sieving, sterilizing at 121 ℃ and then putting the soilless nutrient substrate into a plastic box.
3) Tissue culture: and inserting the soaked root hairs into the soilless culture substrate for 2.5cm, and keeping the distance between the root hairs and the soilless culture substrate for 3 cm. Then, spraying a growth promoting agent to wet the soilless nutrient substrate; carrying out tissue culture in a tissue culture room at the temperature of 27 ℃ and the humidity of 75%; spraying water and lighting during tissue culture.
4) Hardening seedlings: after 15 days or so, transplanting the root hairs with the length of more than 10cm into a black film hardening box with organic fertilizer nutrient soil, and hardening the seedlings in a lighting greenhouse covered with a lighting plastic film; spraying water and shading sun during seedling hardening, and controlling the temperature in the greenhouse to be more than 27 ℃.
5) Transplanting in a field: after hardening off for about 30 days, transplanting the root hair with the length of more than 30cm to a field. Wherein, the growth promoting agent is sprayed on the black film hardening box before transplanting, so that the black film hardening box is thoroughly sprayed.
Comparative example 1
The difference between the comparative example 1 and the example 1 is that the fibrous roots are not soaked in the growth promoter, the tissue culture time is about 30 days, and the hardening period is about 4 months.
Comparative example 2
The comparative example 2 is different from the example 1 in that the fibrous root has a diameter of 5 to 10mm, the tissue culture time is about 20 days, and the hardening period is about 3 months.
Comparative example 3
In the embodiment, the paper mulberry leaf tissue is used as a tissue culture object, the tissue culture mode is a traditional mode, the paper mulberry leaf tissue needs to root and bud again for leaf division in a tissue culture bottle for about 2 months, then the tissue culture seedling is transplanted to a small nutrition box, the seedling is cultivated in a plastic greenhouse, and finally the seedling is hardened to be capable of being planted in a field for about 5 months.
In addition, the method described in the Anhao example 1 of the invention is used for seedling culture in 3/5/2017 and in 3/10/3, respectively, the effect is good, the field growth is good at present, and the root hair tissue culture is proved to be not limited by time and to be fertile all the year round. In addition, the tissue culture seedling culture by adopting the leaf tissue in the past needs about 2 months of rooting, re-sprouting and tillering in a tissue culture bottle, and finally the seedling culture can be moved to a field for planting for about 5 months, so that the time is long, the cost is high, the tissue culture requirement is high, the tissue culture time by adopting the root hair is shortened to 15 days, and the seedling culture in a plastic greenhouse is carried out for about 30 days, so that the seedling culture can be moved to the field for planting. The cost of each seedling of the root hair tissue culture seedling is 0.32 yuan, on one hand, the seedling cost is reduced for high yield of field close planting, and on the other hand, the variety of the paper mulberry can be kept not to be easily degenerated by the root hair tissue culture seedling. More importantly, the seedling culture can be carried out on a large scale all year round to meet the market demand.
The raw materials and equipment used in the invention are common raw materials and equipment in the field if not specified; the methods used in the present invention are conventional in the art unless otherwise specified.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and all simple modifications, alterations and equivalents of the above embodiments according to the technical spirit of the present invention are still within the protection scope of the technical solution of the present invention.
Claims (4)
1. A tissue culture and rapid seedling method for paper mulberry root hairs is characterized by comprising the following steps:
1) cutting root hairs: before excavating a parent broussonetia papyrifera, cutting off branches 35-45cm above the ground; digging up soil of a female broussonetia papyrifera, cleaning root hairs of the female broussonetia papyrifera, cutting the root hairs with the diameter of 1mm into 3-4cm in length in a sterile environment, and soaking the root hairs in a growth promoting agent; after cutting off root hairs, soaking the rest roots in the growth promoting agent for 3-7min, immediately planting and watering again and applying organic fertilizer for daily weeding;
the female broussonetia papyrifera is a space hybrid broussonetia papyrifera variety of Chinese academy of sciences;
the preparation method of the growth promoter comprises the following steps: taking paper mulberry root tissues, crushing, performing biological enzymolysis fermentation to obtain active peptide freeze-dried powder, then preparing into a solution with the mass concentration of 15-25%, and respectively adding 0.1-0.3% of colchicine, 0.5-1.5% of indole auxin and 1-3% of naphthoic acid auxin in the mass of the solution to prepare a growth promoter;
2) preparing a soilless nutrient substrate: the soilless culture substrate comprises the following components in percentage by mass: 8-12% of rapeseed meal powder after fermentation treatment and 8-12% of tea seed meal powder after fermentation treatment; 18-22% of fermented broussonetia papyrifera hard branch powder; 55-65% of vermiculite powder; the thickness of the soilless culture substrate is 5.5-6.5 cm;
3) tissue culture: inserting the soaked root hairs into a soilless culture substrate for 1.5-2.5cm, and keeping the distance between the root hairs and the soilless culture substrate for 2-3 cm; then, spraying a growth promoting agent to wet the soilless nutrient substrate; performing tissue culture in a tissue culture room at 23-27 deg.C and 65-75% humidity; spraying water supply and illumination are carried out during tissue culture;
4) hardening seedlings: after tissue culture is carried out for 13-17 days, transplanting the root hairs with the length of more than 10cm into a black film hardening box with organic fertilizer nutrient soil, and hardening seedlings in a lighting greenhouse covered with a lighting plastic film; spraying water and shading during seedling hardening, and controlling the temperature in the greenhouse to be above 23-27 ℃;
5) transplanting in a field: after hardening seedlings for 28-32 days, transplanting the root hairs with the length of more than 30cm into a field.
2. The tissue culture rapid seedling raising method of broussonetia papyrifera root whiskers as claimed in claim 1, wherein in the step 1), the root whiskers are soaked for 50-70 min.
3. The tissue culture and rapid seedling raising method for the paper mulberry root and beard as claimed in claim 1, wherein in the step 4), when the outside natural temperature is above-10 ℃ in winter, a plastic film small shed is arranged in the greenhouse, the small shed is covered with non-woven fabrics, and foliar fertilizer is sprayed at intervals of 8-12 days.
4. The tissue culture rapid seedling raising method for broussonetia papyrifera root hairs as claimed in claim 1, wherein in step 5), the growth promoting agent is sprayed on the black film hardening box before field transplantation, so that the black film hardening box is thoroughly sprayed.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810419045.0A CN108812260B (en) | 2018-05-04 | 2018-05-04 | Tissue culture and rapid seedling method for paper mulberry root hairs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201810419045.0A CN108812260B (en) | 2018-05-04 | 2018-05-04 | Tissue culture and rapid seedling method for paper mulberry root hairs |
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| CN110603969A (en) * | 2019-10-12 | 2019-12-24 | 金陵科技学院 | Root cutting method for hybrid paper mulberry |
| CN113207690B (en) * | 2021-05-25 | 2022-03-11 | 华南农业大学 | Efficient one-step regeneration method taking paper mulberry root as explant |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101379918A (en) * | 2007-09-05 | 2009-03-11 | 北京万富春森林资源发展有限公司 | Stem cottage propagation tree-reconstruction method |
| CN101720658A (en) * | 2009-12-16 | 2010-06-09 | 曹川� | Method for soilless culture of broussonetia papyrifera seedlings |
| CN102577976A (en) * | 2012-03-17 | 2012-07-18 | 福建农林大学 | Simple tissue culture method for broussonetia papyrifera |
| CN106962054A (en) * | 2017-05-18 | 2017-07-21 | 贵州务川科华生物科技有限公司 | A kind of paper mulberry plantation soil matrix |
| CN107125132A (en) * | 2017-03-27 | 2017-09-05 | 河池乐康生态农业科技有限公司 | One kind hybridization paper mulberry quick breeding method for tissue culture |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101379918A (en) * | 2007-09-05 | 2009-03-11 | 北京万富春森林资源发展有限公司 | Stem cottage propagation tree-reconstruction method |
| CN101720658A (en) * | 2009-12-16 | 2010-06-09 | 曹川� | Method for soilless culture of broussonetia papyrifera seedlings |
| CN102577976A (en) * | 2012-03-17 | 2012-07-18 | 福建农林大学 | Simple tissue culture method for broussonetia papyrifera |
| CN107125132A (en) * | 2017-03-27 | 2017-09-05 | 河池乐康生态农业科技有限公司 | One kind hybridization paper mulberry quick breeding method for tissue culture |
| CN106962054A (en) * | 2017-05-18 | 2017-07-21 | 贵州务川科华生物科技有限公司 | A kind of paper mulberry plantation soil matrix |
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