CN103444526A - Growth-promoting cultivation method for fritillaria unibiacteata - Google Patents
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Abstract
The invention discloses a growth-promoting cultivation method for fritillaria unibiacteata. The growth-promoting cultivation method comprises the steps of selection and disinfection of materials, treatment of the materials, primary culture, secondary culture, rooting culture, greenhouse acclimatization and transplanting of rooting seedlings in test tubes and soil matrix management. According to the growth-promoting cultivation method, fresh fritillaria unibiacteata bulbs are adopted as explants for propagation, the utilization rate of the bulbs is as high as above 95%, and cultivated plants have flourish branches and leaves and developed root systems, and can vigorously grow. One bulb with the diameter of 4cm can propagate about 150-200 small bulbs, so that limited resources can be utilized for propagation, and the propagation coefficient is very high. In the process of the secondary culture, after thin buds are generated through callus differentiation, the culture temperature is raised, the illumination period is increased, the illumination intensity is enhanced, and the soil fertility and the control rate of diseases and pests are improved, so that the culture time is shortened.
Description
Technical field
The present invention relates to the method for tissue culture of a kind of bulb of fritillary, relate in particular to a kind of method for quickly breeding that is applicable to the dark violet bulb of fritillary.
Background technology
The dark violet bulb of fritillary (F. unibiacteata Hsiao et K. C. Hsia) is traditional Chinese medicine famousr and precious in Fritillaria, the effectively cough-relieving of its bulb, eliminate the phlegm, clearing heat and moistening lung, be particularly suitable for the cough of the cureless intractable cold of insufficiency type of the elderly and children.The demand of the dark violet bulb of fritillary on Chinese Medicinal Materials Markets is high especially, but its bulb growth speed is slow especially, grow up to the general needs time of 4 years of commodity medicinal material from seed, due to bulb of fritillary good drug efficacy, be worth high, add that the artificial planting difficulty is large, reproduction coefficient is low, growth cycle is long, therefore the commodity price of medicinal material is always high, and demand is increasing in recent years, causes the price of the dark violet bulb of fritillary also more and more higher.The wild dark violet bulb of fritillary mainly is distributed in the loud and sonorous cold of physical features, the growth of sunshine is strong, air is dry, spring and autumn is of short duration area.Main producing region Ruoergai, Hongyuan two counties are because being affected by landform, cold current etc., throughout the year without the summer, this climatic characteristic has formed the special environment condition of dark violet bulb of fritillary growth, so this special growing environment has determined the dark violet bulb of fritillary in field and artificial planting is more difficult, in order to enlarge the breeding of the dark violet bulb of fritillary, utilize tissue culture technique to carry out Fast-propagation very necessary.The research of the current tissue culture technique to the bulb of fritillary begins to take shape, but is not also a lot of to the research of the dark violet bulb of fritillary, and propagation method does not form a set of complete breeding system also not on a large scale.
Summary of the invention
The technical problem that the present invention mainly solves is to provide a kind of method of utilizing tissue culture technique to breed the dark violet bulb of fritillary, for the fast breeding of the dark violet bulb of fritillary provides a set of complete tissue culture propagating system.
For solving the problems of the technologies described above, the technical scheme that the present invention adopts is:
The growth-promoting cultivation method of a kind of dark violet bulb of fritillary, is characterized in that, comprises the following steps:
(1) selection of material and sterilization: at first select robust growth, ripe fresh dark violet bulb of fritillary bulb ball, peel off scale and the remaining old root of base portion of bulb outermost layer shriveling, flowing water rinses after 30 min to be put into clear water and soaks 15min, with after 500 times of immersion bubble 20-30 min of 50% carbendazim, the strip off in layer by the bulb ball, irradiate again 20 min under uviol lamp, then proceed to superclean bench, first use the alcohol immersion 10 seconds of 70-75 % on superclean bench, then use aseptic water washing, proceed to again in the mercuric chloride solution of 0.1 % that has dripped 2～3 POLYSORBATE 80s and soak 5min, with aseptic water washing 4 times,
(2) processing of material: the scale of disinfecting is blotted with aseptic filter paper, under aseptic condition, also have base section to isolate according to upper, middle and lower section material, be cut into respectively the fritter of 4-6mm;
(3) first culture: the explant under aseptic condition, step (2) obtained is seeded on first culture base, and this medium is at the NAA of the 6-BA, the 1.2mg/L that have added 1.0mg/L in the MS conventional medium, the ZT of 0.4mg/L; Cultivation temperature is 26 ℃, light application time 14 h/d, and intensity of illumination 22 00 Lx, PH is 6.5, agar 5 g/L, through 15-20 days formation callus;
(4) subculture is cultivated: the callus of first culture is cut and is seeded on subculture medium, this medium is the KT of NAA, 0.3mg/L of the 6-BA, 0.4 mg/L that have added 0.5mg/L in the 3/4MS conventional medium and the sucrose of 30mg/L, cultivation temperature is 22 ℃, periodicity of illumination 13h/d, luminous intensity is 2500lux, PH 6.5, after 4-5 days time differentiated thin bud, rising cultivation temperature to 25 ℃, periodicity of illumination is 14h/d, luminous intensity is 2800lux, continues to cultivate the clove that just can differentiate the cluster cluster in 6-10 days;
(5) culture of rootage: the clove that will break up out is inoculated in respectively one by one in following inducing culture and carries out root induction: the IBA of the active carbon of MS medium, 1.5mg/L, the agar of 6g/L, 0.3mg/L, the KT of 0.4mg/L, cultivation temperature is 23 ℃, periodicity of illumination 12h/d, luminous intensity is 2000lux, pH value is 6.0, high through within 20-25 days, treating that clove grows to 3cm, when root system is more healthy and stronger and grow 6-10 root, start the next stage;
(5) soil matrix management: regulating the soil pH value is 7.0～8.2, after mixing with 5 times of volume fine earths after Cosan is sieved, use, except regulating the soil pH value with Cosan, Cosan is better with peat composed of rotten mosses mixing application effect, perhaps use pine needle to coordinate Cosan to be improved soil, wherein pine needle, original soil, muck, Cosan are according to ratio of weight and number 2:3:1:2, perhaps in soil, apply saw not, the peat composed of rotten mosses, liver moss and Cosan carry out soil melioration, wherein saw is not, the ratio of weight and number of the peat composed of rotten mosses, liver moss and Cosan is 1:1:1:2;
Increase soil organic matter content: the field planting ditch that digs wide 50 cm, dark 40 cm, organic matter, fertilizer, original soil are improved the soil in the ratio of 1:1:1, the acid peat composed of rotten mosses of organic matter, if more than must first stacking January with sawdust, bark, become thoroughly decomposed fully after decomposing and re-use, as the employment domestic animals manure becomes thoroughly decomposed or straw leaf rot the to become thoroughly decomposed natural fertilizers of rear formation, note not mixing ash, lime alkaline matter, content reaches more than 3%～5%, to increase the permeability of soil;
Soil covers: soil can carry out after covering seedling planting, by the covering uniform fold at bed surface, wide 1 meter, thick 5～10 centimetres, cover again 2 cm thicks every year later, to keep original thickness, the new sawdust decomposed if application is not rotted, need enrich 50% nitrogenous fertilizer, the sawdust decomposed rots, nitrogen fertilizer amount should correspondingly reduce, cover the unregistered land film and can prevent soil water evaporation, control weeds, improve ground temperature, preferably the orchard application of drip irrigation facility is being arranged, covering the unregistered land film is combined with the organic phase of covering before the better or field planting of effect and kills weeds with weed killer herbicide, entirely cultivate field after field planting and sow green manure Bai Sanye, the leguminous plants such as red clover, can cover field fully after March, can increase soil fertility again,
(6) rooting tube plantlet greenhouse acclimatization and transplants:
By the rearmounted greenhouse temperature of the test-tube plantlet uncork of taking root at 25-28 ℃, humidity 80-85%, intensity of illumination after 3-5 days, is washed the medium on seedling at 5000-5500lx condition lower refining seedling off, and it is transplanted in the soil matrix in above-mentioned steps (5); When extraneous nocturnal temperature at 10-15 ℃, day temperature is transplanted test-tube plantlet between 18-25 the time, in 5 days of the initial stage of transplanting, with plastic film, cover, to keep humidity more than 80%, and controlled light intensity, at 2700-3300lx, is thrown off gradually afterwards film and suitably increases illumination, until transplanted seedling is placed under natural conditions.
The invention has the beneficial effects as follows:
(1) the present invention adopts fresh dark violet bulb of fritillary bulb ball to be bred as explant, and the availability of bulb ball is very high, reach more than 95%, and the plant branches and leaves that cultivate are luxuriant, and well developed root system, plant strain growth is vigorous; A diameter 4cm bulb ball approximately can be bred 150-200 clove, can utilize limited resource to be bred, and reproduction coefficient is very high.
(2) in Subculture after Calli Differentiation goes out thin bud, the rising cultivation temperature, improve periodicity of illumination and intensity of illumination, shortened like this incubation time.
(3) work as extraneous nocturnal temperature at 10-15 ℃ in rooting tube plantlet greenhouse acclimatization and transplants process, day temperature is transplanted between 18-25 the time, be conducive to growing of the dark violet bulb of fritillary, held the time of transplanting, improved soil fertility, controlled damage by disease and insect, the transplanting success rate of test-tube plantlet has been reached more than 95%.
Embodiment
Below preferred embodiment of the present invention is described in detail, thereby so that advantages and features of the invention can be easier to be it will be appreciated by those skilled in the art that, protection scope of the present invention is made to more explicit defining.
embodiment 1
The growth-promoting cultivation method of a kind of dark violet bulb of fritillary comprises:
(1) selection of material and sterilization: at first select robust growth, ripe fresh dark violet bulb of fritillary bulb ball, peel off scale and the remaining old root of base portion of bulb outermost layer shriveling, flowing water rinses after 30 min to be put into clear water and soaks 15min, after steeping 20 min by 500 times of immersions of 50% carbendazim, the strip off in layer by the bulb ball, irradiate again 20 min under uviol lamp, then proceed to superclean bench, first use the alcohol immersion 10 seconds of 70 % on superclean bench, then use aseptic water washing, proceed to again in the mercuric chloride solution of 0.1 % that has dripped 2 POLYSORBATE 80s and soak 5min, with aseptic water washing 4 times,
(2) processing of material: the scale of disinfecting is blotted with aseptic filter paper, under aseptic condition, also have base section to isolate according to upper, middle and lower section material, be cut into respectively the fritter of 4mm;
(3) first culture: the explant under aseptic condition, step (2) obtained is seeded on first culture base, and this medium is at the NAA of the 6-BA, the 1.2mg/L that have added 1.0mg/L in the MS conventional medium, the ZT of 0.4mg/L; Cultivation temperature is 26 ℃, light application time 14 h/d, and intensity of illumination 22 00 Lx, PH is 6.5, agar 5 g/L, through 15-20 days formation callus;
(4) subculture is cultivated: the callus of first culture is cut and is seeded on subculture medium, this medium is the KT of NAA, 0.3mg/L of the 6-BA, 0.4 mg/L that have added 0.5mg/L in the 3/4MS conventional medium and the sucrose of 30mg/L, cultivation temperature is 22 ℃, periodicity of illumination 13h/d, luminous intensity is 2500lux, PH 6.5, after 4 day time differentiated thin bud, rising cultivation temperature to 25 ℃, periodicity of illumination is 14h/d, luminous intensity is 2800lux, continues to cultivate the clove that just can differentiate the cluster cluster in 8 days;
(5) culture of rootage: the clove that will break up out is inoculated in respectively one by one in following inducing culture and carries out root induction: the IBA of the active carbon of MS medium, 1.5mg/L, the agar of 6g/L, 0.3mg/L, the KT of 0.4mg/L, cultivation temperature is 23 ℃, periodicity of illumination 12h/d, luminous intensity is 2000lux, pH value is 6.0, high through within 20-25 days, treating that clove grows to 3cm, when root system is more healthy and stronger and grow 6-10 root, start the next stage;
(6) rooting tube plantlet greenhouse acclimatization and transplants: by the rearmounted greenhouse temperature of the test-tube plantlet uncork of taking root at 25 ℃, humidity 80%, intensity of illumination is at 5000lx condition lower refining seedling after 5 days, wash the medium on seedling off, by its seedbed that to be transplanted to by sheep excrement, humus soil, perlitic mixed proportion be 1: 3: 2.When extraneous nocturnal temperature at 10-15 ℃, day temperature is transplanted test-tube plantlet between 18-25 the time, in 5 days of the initial stage of transplanting, with plastic film, cover, to keep humidity more than 80%, and controlled light intensity, at 2700lx, is thrown off gradually afterwards film and suitably increases illumination, until transplanted seedling is placed under natural conditions.
Through the cultivation of above-mentioned steps, the callus formation rate of the dark violet bulb of fritillary has reached 95%, and the propagation multiple has reached 5.8 times, and rooting rate has reached 95%, and final survival rate has reached 88%, and the Quality and yield of the dark violet bulb of fritillary all is greatly improved.
embodiment 2
The growth-promoting cultivation method of a kind of dark violet bulb of fritillary comprises the following steps:
(1) selection of material and sterilization: at first select robust growth, ripe fresh dark violet bulb of fritillary bulb ball, peel off scale and the remaining old root of base portion of bulb outermost layer shriveling, flowing water rinses after 30 min to be put into clear water and soaks 15min, after steeping 30 min by 500 times of immersions of 50% carbendazim, the strip off in layer by the bulb ball, irradiate again 20 min under uviol lamp, , then proceed to superclean bench, first use the alcohol immersion 10 seconds of 75 % on superclean bench, then use aseptic water washing, proceed to again in the mercuric chloride solution of 0.1 % that has dripped 3 POLYSORBATE 80s and soak 5min, with aseptic water washing 4 times,
(2) processing of material: the scale of disinfecting is blotted with aseptic filter paper, under aseptic condition, also have base section to isolate according to upper, middle and lower section material, be cut into respectively the fritter of 6mm;
(3) first culture: the explant under aseptic condition, step (2) obtained is seeded on first culture base, and this medium is at the NAA of the 6-BA, the 1.2mg/L that have added 1.0mg/L in the MS conventional medium, the ZT of 0.4mg/L; Cultivation temperature is 26 ℃, light application time 14 h/d, and intensity of illumination 22 00 Lx, PH is 6.5, agar 5 g/L, through 20 days formation callus;
(4) subculture is cultivated: the callus of first culture is cut and is seeded on subculture medium, this medium is the KT of NAA, 0.3mg/L of the 6-BA, 0.4 mg/L that have added 0.5mg/L in the 3/4MS conventional medium and the sucrose of 30mg/L, cultivation temperature is 22 ℃, periodicity of illumination 13h/d, luminous intensity is 2500lux, PH 6.5, after 4-5 days time differentiated thin bud, rising cultivation temperature to 25 ℃, periodicity of illumination is 14h/d, luminous intensity is 2800lux, continues to cultivate the clove that just can differentiate the cluster cluster in 6-10 days;
(5) culture of rootage: the clove that will break up out is inoculated in respectively one by one in following inducing culture and carries out root induction: the IBA of the active carbon of MS medium, 1.5mg/L, the agar of 6g/L, 0.3mg/L, the KT of 0.4mg/L, cultivation temperature is 23 ℃, periodicity of illumination 12h/d, luminous intensity is 2000lux, pH value is 6.0, high through within 20-25 days, treating that clove grows to 3cm, when root system is more healthy and stronger and grow 6-10 root, start the next stage;
(6) rooting tube plantlet greenhouse acclimatization and transplants: by the rearmounted greenhouse temperature of the test-tube plantlet uncork of taking root at 28 ℃, humidity 85%, intensity of illumination is at 5500lx condition lower refining seedling after 3 days, wash the medium on seedling off, by its seedbed that to be transplanted to by sheep excrement, humus soil, perlitic mixed proportion be 1: 3: 2.When extraneous nocturnal temperature at 15 ℃, day temperature is transplanted test-tube plantlet between 25 the time, in 5 days of the initial stage of transplanting, with plastic film, cover, to keep humidity more than 80%, and controlled light intensity, at 3300lx, is thrown off gradually afterwards film and suitably increases illumination, until transplanted seedling is placed under natural conditions.
Through the cultivation of above-mentioned steps, the callus formation rate of the dark violet bulb of fritillary has reached 92%, and the propagation multiple has reached 6 times, and rooting rate has reached 98%, and final survival rate has reached 90%.
(5) soil matrix management: regulating the soil pH value is 7.0～8.2, after mixing with 5 times of volume fine earths after Cosan is sieved, use, except regulating the soil pH value with Cosan, Cosan is better with peat composed of rotten mosses mixing application effect, perhaps use pine needle to coordinate Cosan to be improved soil, wherein pine needle, original soil, muck, Cosan are according to ratio of weight and number 2:3:1:2, perhaps in soil, apply saw not, the peat composed of rotten mosses, liver moss and Cosan carry out soil melioration, wherein saw is not, the ratio of weight and number of the peat composed of rotten mosses, liver moss and Cosan is 1:1:1:2;
Increase soil organic matter content: the field planting ditch that digs wide 50 cm, dark 40 cm, organic matter, fertilizer, original soil are improved the soil in the ratio of 1:1:1, the acid peat composed of rotten mosses of organic matter, if more than must first stacking January with sawdust, bark, become thoroughly decomposed fully after decomposing and re-use, as the employment domestic animals manure becomes thoroughly decomposed or straw leaf rot the to become thoroughly decomposed natural fertilizers of rear formation, note not mixing ash, lime alkaline matter, content reaches more than 3%～5%, to increase the permeability of soil;
Soil covers: soil can carry out after covering seedling planting, by the covering uniform fold at bed surface, wide 1 meter, thick 5～10 centimetres, cover again 2 cm thicks every year later, to keep original thickness, the new sawdust decomposed if application is not rotted, need enrich 50% nitrogenous fertilizer, the sawdust decomposed rots, nitrogen fertilizer amount should correspondingly reduce, cover the unregistered land film and can prevent soil water evaporation, control weeds, improve ground temperature, preferably the orchard application of drip irrigation facility is being arranged, covering the unregistered land film is combined with the organic phase of covering before the better or field planting of effect and kills weeds with weed killer herbicide, entirely cultivate field after field planting and sow green manure Bai Sanye, the leguminous plants such as red clover, can cover field fully after March, can increase soil fertility again,
(6) rooting tube plantlet greenhouse acclimatization and transplants:
By the rearmounted greenhouse temperature of the test-tube plantlet uncork of taking root at 25-28 ℃, humidity 80-85%, intensity of illumination after 3-5 days, is washed the medium on seedling at 5000-5500lx condition lower refining seedling off, and it is transplanted in the soil matrix in above-mentioned steps (5); When extraneous nocturnal temperature at 10-15 ℃, day temperature is transplanted test-tube plantlet between 18-25 the time, in 5 days of the initial stage of transplanting, with plastic film, cover, to keep humidity more than 80%, and controlled light intensity, at 2700-3300lx, is thrown off gradually afterwards film and suitably increases illumination, until transplanted seedling is placed under natural conditions.
Through the cultivation of above-mentioned steps, the callus formation rate of the dark violet bulb of fritillary has reached 94%, and the propagation multiple has reached 7 times, and rooting rate has reached 99%, and final survival rate has reached 98%.
The foregoing is only embodiments of the invention; not thereby limit the scope of the claims of the present invention; every equivalent structure or conversion of equivalent flow process that utilizes description of the present invention to do; or directly or indirectly be used in other relevant technical fields, all in like manner be included in scope of patent protection of the present invention.
Claims (1)
1.
the growth-promoting cultivation method of a kind of dark violet bulb of fritillary, is characterized in that, comprises the following steps:
(1) selection of material and sterilization: at first select robust growth, ripe fresh dark violet bulb of fritillary bulb ball, peel off scale and the remaining old root of base portion of bulb outermost layer shriveling, flowing water rinses after 30 min to be put into clear water and soaks 15min, with after 500 times of immersion bubble 20-30 min of 50% carbendazim, the strip off in layer by the bulb ball, irradiate again 20 min under uviol lamp, then proceed to superclean bench, first use the alcohol immersion 10 seconds of 70-75 % on superclean bench, then use aseptic water washing, proceed to again in the mercuric chloride solution of 0.1 % that has dripped 2～3 POLYSORBATE 80s and soak 5min, with aseptic water washing 4 times,
(2) processing of material: the scale of disinfecting is blotted with aseptic filter paper, under aseptic condition, also have base section to isolate according to upper, middle and lower section material, be cut into respectively the fritter of 4-6mm;
(3) first culture: the explant under aseptic condition, step (2) obtained is seeded on first culture base, and this medium is at the NAA of the 6-BA, the 1.2mg/L that have added 1.0mg/L in the MS conventional medium, the ZT of 0.4mg/L; Cultivation temperature is 26 ℃, light application time 14 h/d, and intensity of illumination 22 00 Lx, PH is 6.5, agar 5 g/L, through 15-20 days formation callus;
(4) subculture is cultivated: the callus of first culture is cut and is seeded on subculture medium, this medium is the KT of NAA, 0.3mg/L of the 6-BA, 0.4 mg/L that have added 0.5mg/L in the 3/4MS conventional medium and the sucrose of 30mg/L, cultivation temperature is 22 ℃, periodicity of illumination 13h/d, luminous intensity is 2500lux, PH 6.5, after 4-5 days time differentiated thin bud, rising cultivation temperature to 25 ℃, periodicity of illumination is 14h/d, luminous intensity is 2800lux, continues to cultivate the clove that just can differentiate the cluster cluster in 6-10 days;
(5) culture of rootage: the clove that will break up out is inoculated in respectively one by one in following inducing culture and carries out root induction: the KT of the IBA+0.4mg/L of agar+0.3mg/L of active carbon+6g/L of MS medium+1.5mg/L, cultivation temperature is 23 ℃, periodicity of illumination 12h/d, luminous intensity is 2000lux, pH value is 6.0, high through within 20-25 days, treating that clove grows to 3cm, when root system is more healthy and stronger and grow 6-10 root, start the next stage;
(6) soil matrix management: regulating the soil pH value is 7.0～8.2, after mixing with 5 times of volume fine earths after Cosan is sieved, use, except regulating the soil pH value with Cosan, Cosan is better with peat composed of rotten mosses mixing application effect, perhaps use pine needle to coordinate Cosan to be improved soil, wherein pine needle, original soil, muck, Cosan are according to ratio of weight and number 2:3:1:2, perhaps in soil, apply saw not, the peat composed of rotten mosses, liver moss and Cosan carry out soil melioration, wherein saw is not, the ratio of weight and number of the peat composed of rotten mosses, liver moss and Cosan is 1:1:1:2;
increase soil organic matter content: the field planting ditch that digs wide 50 cm, dark 40 cm, organic matter, fertilizer, original soil are improved the soil in the ratio of 1:1:1, the acid peat composed of rotten mosses of organic matter, if more than must first stacking January with sawdust, bark, become thoroughly decomposed fully after decomposing and re-use, as the employment domestic animals manure becomes thoroughly decomposed or straw leaf rot the to become thoroughly decomposed natural fertilizers of rear formation, note not mixing ash, lime alkaline matter, content reaches more than 3%～5%, to increase the permeability of soil;
soil covers: soil can carry out after covering seedling planting, by the covering uniform fold at bed surface, wide 1 meter, thick 5～10 centimetres, cover again 2 cm thicks every year later, to keep original thickness, the new sawdust decomposed if application is not rotted, need enrich 50% nitrogenous fertilizer, the sawdust decomposed rots, nitrogen fertilizer amount should correspondingly reduce, cover the unregistered land film and can prevent soil water evaporation, control weeds, improve ground temperature, preferably the orchard application of drip irrigation facility is being arranged, covering the unregistered land film is combined with the organic phase of covering before the better or field planting of effect and kills weeds with weed killer herbicide, entirely cultivate field after field planting and sow green manure Bai Sanye, the leguminous plants such as red clover, can cover field fully after March, can increase soil fertility again,
(7) rooting tube plantlet greenhouse acclimatization and transplants:
by the rearmounted greenhouse temperature of the test-tube plantlet uncork of taking root at 25-28 ℃, humidity 80-85%, intensity of illumination after 3-5 days, is washed the medium on seedling at 5000-5500lx condition lower refining seedling off, and it is transplanted in the soil matrix in above-mentioned steps (5); When extraneous nocturnal temperature at 10-15 ℃, day temperature is transplanted test-tube plantlet between 18-25 the time, in 5 days of the initial stage of transplanting, with plastic film, cover, to keep humidity more than 80%, and controlled light intensity, at 2700-3300lx, is thrown off gradually afterwards film and suitably increases illumination, until transplanted seedling is placed under natural conditions.
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| CN109757377A (en) * | 2019-03-01 | 2019-05-17 | 南京工业大学大丰海洋产业研究院 | A kind of cultural method for accelerating fritillaria thunbergii reproduction speed |
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