CN104145818B - A kind of store method of flameray gerbera germ plasm resource - Google Patents
A kind of store method of flameray gerbera germ plasm resource Download PDFInfo
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Abstract
The present invention discloses a kind of store method of flameray gerbera germ plasm resource, the method comprises selects ligulate flower to be just open into the flameray gerbera flower that tubular flower has told powder, stay the long 5cm ~ 6cm of bennet, explant after treatment passes through Fiber differentiation, the continuous squamous subculture of subculture medium of two different formulations, in booth after culture of rootage, Seedling bag is cultivated and is planted 1 year in canopy.A retention cycle can be preserved flameray gerbera germ plasm resource and reach 3 ~ 4 years, the field survival rate more than 90% of Germplasms.The method preserves flameray gerbera provenances by rejuvenation two links in the squamous subculture successively of two kinds of different subculture mediums and canopy, reduce seedling variation and quality deterioration problem that the reason such as Hormone Factors, planting environment causes, stably saving the merit of flameray gerbera quality resource, is a kind of newly, effective flameray gerbera germplasm resource preservation method.
Description
Technical field
The present invention relates to a kind of store method of flameray gerbera germ plasm resource, belong to biological technical field.
Background technology
Flameray gerbera (Gerbera jamesonil Bolus) has another name called African daisy, the evergreen herbage flower of composite family Gerbera perennial root.Because its flower is very large, pattern enriches, and attitude is different, by force ornamental, resistance to long-distance transport, deeply favors by consumers in general.Under optimum conditions can whole year production, the anniversary blooms, and can be market supply fresh cut-flowers the whole year, is one of large cut-flower in the world five.China is once a large amount of from Dutch introducing and planting since the nineties in last century, through testing for many years, due to the restriction of weather conditions, culture technique, generally there is quality deterioration phenomenon, good strains of seeds serious degradation is even lost, and loses culture and utility and is worth, and has to progressively eliminate from production.Flameray gerbera deterioration of variety is usually expressed as that flower diminishes, intermediate color is many, flower pattern is disorderly, two stalk phenomenon, plant height are uneven, vitality declines, resistance and output reduction etc.
Cause the reason of flameray gerbera deterioration of variety mainly: planting environment such as illumination, moisture, temperature, nutrition are not suitable for flameray gerbera and grow, the pathogen in soil compaction and salination, continuous cropping soil accumulates and compound dip-dye etc. in a large number, causes kind to lose original typicalness; In flameray gerbera tissue culture procedures, the factors such as the subculture algebraically of cultivation temperature, propagation bud and hormone concentration cause flameray gerbera seedling that asexual variation occurs.So-called kind matter, refer to and determine organism genetic character and genetic material hereditary information being passed to offspring from parental generation, the investigation of germ plasm resource, collection, preservation and evaluation are the bases that scientific development utilizes plant, have conclusive effect to playing germ plasm resource Exploitative potential to greatest extent.
At present, the preservation of flameray gerbera germ plasm resource is preserved mainly with the training of indoor group and field cultivation greatly, tissue culture method mainly using young flower bud holder as the explant of flameray gerbera tissue cultures, but young flower bud holder to there is surface sterilization as explant more difficult, it is large to consume examination material amount, and the size requirements of bud is strict, too small easy death, excessive easy dye rate, Material selec-tion is by time restriction, in field, the same time draws materials, and meets the requirements of the problem that bud is less; The deficiency that current flameray gerbera tissue cultures is preserved shows that the squamous subculture cycle is shorter, and conservation cost is relatively large; The deficiency that in canopy, cultivation is preserved shows that floor space is many, and light, temperature, water and Pest management are more difficult, and the soil compaction of cultivation formation for many years, salination, pathogen accumulate in a large number, and continuous cropping obstacle causes kind to lose original typicalness.Therefore a kind of suitable industry development of flameray gerbera germplasm resource preservation method to flameray gerbera is significant.
Summary of the invention
The defect existed for the preservation aspect of current flameray gerbera germ plasm resource or deficiency, of the present inventionly provide a kind of flameray gerbera germplasm resource preservation method, object is to make that its explant is drawn materials conveniently, subculture cycle lengthens, retain costs reduces, variation reduces, can distinct variety kind preferably, meet the needs that scientific development utilizes flameray gerbera germ plasm resource.
The store method of a kind of flameray gerbera germ plasm resource provided by the present invention, is characterized in that step is as follows:
(1) selection of explant material and pretreatment
1. select anosis worm to endanger, plant the healthy and strong plant of the matter flameray gerbera consistent with parent, win ligulate flower and be just open into the flameray gerbera flower that tubular flower has told powder, stay the long 5cm ~ 6cm of bennet, insert and be equipped with in 5 ~ 15mg/L 6-BA aqueous solution, put into the refrigerator low temperature treatment 3 ~ 7 days of 4 DEG C;
2. material step (1) 1. processed is first with the cleaning of washing powder water, after rinsing well with clear water again, under super-clean environment, with blade excision bennet and ligulate flower edge, first put into the mercuric chloride solution sterilization 15min that mass percentage concentration is 0.1%, being placed in mass percentage concentration is again the 10min that sterilizes in the mixed liquor adding 2 polysorbas20s in the liquor natrii hypochloritis of 2%, use aseptic water washing afterwards 3 ~ 4 times, epiclinal all ligulate flowers are taken out again with tweezers, required flameray gerbera explant is with the thickness of 0.2 ~ 0.3cm bottom scalpel excision calyx edge and holder,
(2) Fiber differentiation
The flameray gerbera explant that (1) is 2. described puts into following media:
MS basic culture solution
Be 1500 ~ 2000lx in intensity of illumination, temperature is 25 ± 2 DEG C, and light application time is under the condition of 10 ~ 12h/d, and Fiber differentiation 25 ~ 35d sprouts to explant and differentiates the high young shoot of 1 ~ 2cm, cuts young shoot;
(3) first time squamous subculture
The young shoot that step (2) cuts being cut into individual plant is transferred in following subculture medium I:
MS Media modified
Described MS Media modified is: in conventional MS culture fluid, the concentration of macroelement reduces by half, and thiamine hydrochloride concentration is 2mg/L, and puridoxine hydrochloride concentration is the culture fluid of 1mg/L;
Be 1500 ~ 2000lx in intensity of illumination, temperature is 18 ± 2 DEG C, and light application time is cultivate 45 ~ 55d under the condition of 10 ~ 12h/d;
(4) second time squamous subculture
The young shoot of step (3) first time squamous subculture is cut into individual plant to be transferred in following subculture medium II:
MS Media modified
Described MS Media modified is identical with the MS Media modified in step (3) first time squamous subculture subculture medium I;
Be 1500 ~ 2000lx in intensity of illumination, temperature is 18 ± 2 DEG C, and light application time is cultivate 60 ~ 70d under the condition of 10 ~ 12h/d;
(5) young shoot of step (4) second time squamous subculture is cut into individual plant to repeat again to use (3) squamous subculture and step (4) second time squamous subculture to 2 ~ 3 year for the first time successively;
(6) the healthy and strong plant of more than the Multiple Buds 2cm of step (5) is cut into individual plant to be inoculated in following root media and to carry out culture of rootage:
1/2MS basic culture solution
Described 1/2MS basic culture solution is: the culture fluid that in conventional MS culture fluid, the concentration of full dose element reduces by half;
At the bottleneck parcel sealed membrane of tissue culture bottle, move to be stamped and booth seedbed that shading rate is 70% sunshade net is cultured to plant base portion grows 3 ~ 5 roots, the temperature of booth controls to be 18 DEG C-28 DEG C;
(7) plant that takes root of step (6) is taken out, clean the medium on seedling routinely, putting into mass percentage concentration is after the carbendazim solution sterilization 1 ~ 2min of 0.1%, transplant in the Seedling bag that mixed-matrix is housed, carry out routinely watering, fertilizing management, after growth 45d, obtain flameray gerbera transplanted seedling; Described mixed-matrix is for being that 2 ︰ 1 ︰ 0.5 are formulated by the perlitic volume ratio of rotten Zhi Tu ︰ Hong Tu ︰;
(8) transplanted seedling that step (7) obtains is planted 1 year in canopy.
Further, according to the utilization plan of flameray gerbera germ plasm resource, above-mentioned steps (1) to step (8) can be repeated and continue to preserve flameray gerbera germ plasm resource.
Compared with prior art, the invention has the beneficial effects as follows:
1, the present invention has just been open into flameray gerbera flower that tubular flower told powder for explant material with ligulate flower and has carried out tissue cultures, decrease injury to maternal plant (because flameray gerbera can be bloomed in the anniversary, and the florescence is longer, reduce the injury to maternal plant, the loss of flameray gerbera commodity can be reduced), and it is more flexible to make explant draw materials, originate more wide, on desirable market, the flower of the fine quality of popular sale is as explant, for the kind matter introduction in breeding work and preservation provide new way.
2, the picking out of excess stock in whole explant disinfecting process, improves sterilization rate effect; The explant got is comparatively ripe, decreases melting brown rate and the lethality of explant.
3, the key link of whole flameray gerbera Germ-plasma resources protection is improved.Explant, by low temperature treatment and HORMONE TREATMENT, utilizes MS Media modified, reduces the consumption of hormone, and young shoot divergaence time only needs 25 ~ 35d, and the minimizing of hormone dosage simultaneously reduces the probability that indefinite bud morphs.
4, the culture of rootage step under conventional chambers fluorescent lamp is changed into cultivate on booth seedbed, not only saved the grid electricity fee cost in culture of rootage stage, and improve seedling of taking root and transplant the adaptive capacity of cultivating to follow-up; Sterile rootage seedling direct transplantation enters in Seedling bag, eliminates the link that plantlet in vitro nutritive cube is transplanted, and the seedling of transplant survival is convenient to transport, directly plants after plucking Seedling bag.
5, the inventive method carries out squamous subculture successively with two kinds of different subculture mediums successively, and not only culture effect is good, and the time of twice squamous subculture adds up to and is 105 days ~ 125 days, reduces conservation cost.
6 the inventive method are preserved flameray gerbera germ plasm resource and can be reached more than 3 ~ 4 years, the field survival rate more than 90% of Germplasms.Flameray gerbera provenances are preserved by rejuvenation two links in the squamous subculture successively of the different subculture medium of the present invention two kinds and canopy, reduce seedling variation and quality deterioration problem that the reason such as Hormone Factors, planting environment causes, reduce simple group training store method relaying and count the variation too much caused from generation to generation, compensate for the weak point of current flameray gerbera Germ-plasma resources protection, stably saving the merit of flameray gerbera quality resource, is a kind of newly, effective flameray gerbera germplasm resource preservation method.
Embodiment
In order to better the present invention is described, provide embodiments of the invention below, but content of the present invention is not limited in this.Without specified otherwise in each embodiment is conventional method.
Embodiment 1
(1) selection of explant material and pretreatment
1. select the harm of plant type better, anosis worm, plant the healthy and strong plant of the matter flameray gerbera consistent with parent, win the flameray gerbera flower that ligulate flower has just opened, stay the long 5cm ~ 6cm of bennet, insertion is equipped with in the 5mg/L 6-BA aqueous solution (namely 6-BA running water is mixed with concentration is the 5mg/L aqueous solution), puts into the refrigerator low temperature treatment 7 days of 4 DEG C;
2. material step (1) 1. processed is first with the cleaning of washing powder water, after rinsing well with clear water again, under super-clean environment, with blade excision bennet and ligulate flower edge (making the size of material be suitable for sterilizing in a reservoir), first put into the mercuric chloride solution sterilization 15min that mass percentage concentration is 0.1%, being placed in mass percentage concentration is again the 10min that sterilizes in the mixed liquor adding 2 polysorbas20s in the liquor natrii hypochloritis of 2%, use aseptic water washing afterwards 3 ~ 4 times, epiclinal all ligulate flowers are taken out again with tweezers, required flameray gerbera explant is with the thickness of 0.2 ~ 0.3 cm bottom scalpel excision calyx edge and holder.
(2) Fiber differentiation
The flameray gerbera explant that (1) is 2. described puts into following media:
MS basic culture solution
Be 1500 ~ 2000lx in intensity of illumination, temperature is 25 ± 2 DEG C, and light application time is under the condition of 10 ~ 12h/d, and Fiber differentiation 25d sprouts to explant and differentiates the high young shoot of 1 ~ 2cm, cuts young shoot.
(3) first time squamous subculture
The young shoot that step (2) cuts being cut into individual plant is transferred in following subculture medium I:
MS Media modified
Described MS Media modified is: in conventional MS culture fluid, the concentration of macroelement reduces by half, and thiamine hydrochloride concentration is 2mg/L, and puridoxine hydrochloride concentration is the culture fluid of 1mg/L;
Be 1500 ~ 2000lx in intensity of illumination, temperature is 18 ± 2 DEG C, and light application time is cultivate 55d under the condition of 10 ~ 12h/d.
(4) second time squamous subculture
The young shoot of step (3) first time squamous subculture is cut into individual plant to be transferred in following subculture medium II:
MS Media modified
Described MS Media modified is identical with the MS Media modified in step (3) first time squamous subculture subculture medium I;
Be 1500 ~ 2000lx in intensity of illumination, temperature is 18 ± 2 DEG C, and light application time is cultivate 70d under the condition of 10 ~ 12h/d.
(5) young shoot of step (4) second time squamous subculture is cut into individual plant to repeat again to use (3) squamous subculture and step (4) second time squamous subculture to 2 ~ 3 year for the first time successively.
(6) the healthy and strong plant of more than the Multiple Buds 2cm of step (5) is cut into individual plant to be inoculated in following root media and to carry out culture of rootage:
1/2 MS basic culture solution
Described 1/2MS basic culture solution is: the culture fluid that in conventional MS culture fluid, the concentration of full dose element reduces by half;
At the bottleneck parcel sealed membrane of tissue culture bottle, move to be stamped and booth seedbed that shading rate is 70% sunshade net is cultured to plant base portion grows 3 ~ 5 roots, the temperature of booth controls to be 18 DEG C-28 DEG C.
(7) plant that takes root of step (6) is taken out, clean the medium on seedling routinely, putting into mass percentage concentration is after the carbendazim solution sterilization 1 ~ 2min of 0.1%, transplant in the Seedling bag that mixed-matrix is housed, carry out routinely watering, fertilizing management, after growth 45d, obtain flameray gerbera transplanted seedling; Described mixed-matrix is for being that 2 ︰ 1 ︰ 0.5 are formulated by the perlitic volume ratio of rotten Zhi Tu ︰ Hong Tu ︰;
(8) transplanted seedling that step (7) obtains is planted 1 year in canopy.
Embodiment 2
Embodiment 2 is divided by outside lower operation difference, and all the other operations are identical with embodiment 1, repeat no more.
(1) selection of explant material and pretreatment
1. select the harm of plant type better, anosis worm, plant the healthy and strong plant of the matter flameray gerbera consistent with parent, win ligulate flower opening completely, but tubular flower does not also tell the flameray gerbera flower of powder, staying bennet long is 5cm ~ 6cm, insertion is equipped with in the 10mg/L 6-BA aqueous solution, puts into the refrigerator low temperature treatment 5 days of 4 DEG C;
(2) Fiber differentiation
Inducing culture:
MS basic culture solution
(3) first time squamous subculture
Subculture medium I:
MS Media modified
(4) second time squamous subculture
Subculture medium II:
MS Media modified
Embodiment 3
Embodiment 3 is divided by outside lower operation difference, and all the other operations are identical with embodiment 1, repeat no more.
(1) selection of explant material and pretreatment
1. select the harm of plant type better, anosis worm, plant the healthy and strong plant of the matter flameray gerbera consistent with parent, win the flameray gerbera flower that tubular flower has told powder, stay bennet long to be 5cm ~ 6cm, to insert and be equipped with in the 15mg/L6-BA aqueous solution, put into the refrigerator low temperature treatment 3 days of 4 DEG C;
(2) Fiber differentiation
Inducing culture:
MS basic culture solution
(3) first time squamous subculture
Subculture medium I:
MS Media modified
(4) second time squamous subculture
Subculture medium II:
MS Media modified
Each embodiment preservation flameray gerbera germ plasm resource can reach more than 3 ~ 4 years above, the field survival rate more than 90% of Germplasms.Flameray gerbera provenances are preserved by rejuvenation two links in the squamous subculture successively of the different subculture medium of the present invention two kinds and canopy, reduce seedling variation and quality deterioration problem that the reason such as Hormone Factors, planting environment causes, reduce simple group training store method relaying and count the variation too much caused from generation to generation, compensate for the weak point of current flameray gerbera Germ-plasma resources protection, stably saving the merit of flameray gerbera quality resource, is a kind of newly, effective flameray gerbera germplasm resource preservation method.
Claims (1)
1. a store method for flameray gerbera germ plasm resource, is characterized in that step is as follows:
(1) selection of explant material and pretreatment
1. select anosis worm to endanger, plant the healthy and strong plant of the matter flameray gerbera consistent with parent, win ligulate flower and be just open into the flameray gerbera flower that tubular flower has told powder, stay the long 5cm ~ 6cm of bennet, insert and be equipped with in 5 ~ 15mg/L 6-BA aqueous solution, put into the refrigerator low temperature treatment 3 ~ 7 days of 4 DEG C;
2. material step (1) 1. processed is first with the cleaning of washing powder water, after rinsing well with clear water again, under super-clean environment, with blade excision bennet and ligulate flower edge, first put into the mercuric chloride solution sterilization 15min that mass percentage concentration is 0.1%, being placed in mass percentage concentration is again the 10min that sterilizes in the mixed liquor adding 2 polysorbas20s in the liquor natrii hypochloritis of 2%, use aseptic water washing afterwards 3 ~ 4 times, epiclinal all ligulate flowers are taken out again with tweezers, required flameray gerbera explant is with the thickness of 0.2 ~ 0.3cm bottom scalpel excision calyx edge and holder,
(2) Fiber differentiation
The flameray gerbera explant that (1) is 2. described puts into following media:
MS basic culture solution
Be 1500 ~ 2000lx in intensity of illumination, temperature is 25 ± 2 DEG C, and light application time is under the condition of 10 ~ 12h/d, and Fiber differentiation 25 ~ 35d sprouts to explant and differentiates the high young shoot of 1 ~ 2cm, cuts young shoot;
(3) first time squamous subculture
The young shoot that step (2) cuts being cut into individual plant is transferred in following subculture medium I:
MS Media modified
Described MS Media modified is: in conventional MS culture fluid, the concentration of macroelement reduces by half, and thiamine hydrochloride concentration is 2mg/L, and puridoxine hydrochloride concentration is the culture fluid of 1mg/L;
Be 1500 ~ 2000lx in intensity of illumination, temperature is 18 ± 2 DEG C, and light application time is cultivate 45 ~ 55d under the condition of 10 ~ 12h/d;
(4) second time squamous subculture
The young shoot of step (3) first time squamous subculture is cut into individual plant to be transferred in following subculture medium II:
MS Media modified
Described MS Media modified is identical with the MS Media modified in step (3) first time squamous subculture subculture medium I;
Be 1500 ~ 2000lx in intensity of illumination, temperature is 18 ± 2 DEG C, and light application time is cultivate 60 ~ 70d under the condition of 10 ~ 12h/d;
(5) young shoot of step (4) second time squamous subculture is cut into individual plant to repeat again to use (3) squamous subculture and step (4) second time squamous subculture to 2 ~ 3 year for the first time successively;
(6) the healthy and strong plant of more than the Multiple Buds 2cm of step (5) is cut into individual plant to be inoculated in following root media and to carry out culture of rootage:
1/2MS basic culture solution
Described 1/2MS basic culture solution is: the culture fluid that in conventional MS culture fluid, the concentration of full dose element reduces by half;
At the bottleneck parcel sealed membrane of tissue culture bottle, move to be stamped and booth seedbed that shading rate is 70% sunshade net is cultured to plant base portion grows 3 ~ 5 roots, the temperature of booth controls to be 18 DEG C-28 DEG C;
(7) plant that takes root of step (6) is taken out, clean the medium on seedling routinely, putting into mass percentage concentration is after the carbendazim solution sterilization 1 ~ 2min of 0.1%, transplant in the Seedling bag that mixed-matrix is housed, carry out routinely watering, fertilizing management, after growth 45d, obtain flameray gerbera transplanted seedling; Described mixed-matrix is by humus soil: laterite: perlitic volume ratio is that 2:1:0.5 is formulated;
(8) transplanted seedling that step (7) obtains is planted 1 year in canopy.
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| CN104719161B (en) * | 2015-03-23 | 2017-04-12 | 上海市农业科学院 | Method for obtaining African daisy regeneration plant through inducing somatic embryo |
| CN105123524B (en) * | 2015-09-10 | 2017-09-12 | 新沂时集创新创业科技产业园有限公司 | A kind of preserving seed method of dahlia tissue culture propagation |
| CN105104202B (en) * | 2015-09-10 | 2017-10-20 | 新沂市合沟工业集中区建设发展有限公司 | A kind of preserving seed method of African Chrysanthemum tissue culture propagation |
| CN105123526B (en) * | 2015-09-10 | 2017-05-17 | 广东百林园林股份有限公司 | Germplasm storage method of zinnia elegan tissue culture propagation |
| CN106106158A (en) * | 2016-06-29 | 2016-11-16 | 无锡南理工科技发展有限公司 | A kind of store method of African Chrysanthemum germ plasm resource |
| CN109964816A (en) * | 2019-04-15 | 2019-07-05 | 云南省农业科学院花卉研究所 | A kind of method for culturing seedlings of African Chrysanthemum monoploid transplanted seedling |
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| CN101361456A (en) * | 2008-07-25 | 2009-02-11 | 云南省农业科学院花卉研究所 | Efficient flameray-gerbera propagation production method using excised leaf |
| CN101720662A (en) * | 2009-12-30 | 2010-06-09 | 云南省农业科学院花卉研究所 | Cultivating method of ball-shaped gerbera coenospecies |
| CN102657094B (en) * | 2012-05-24 | 2013-11-20 | 云南省农业科学院花卉研究所 | Ex-vitro soilless cutting rooting method for tissue-cultured and proliferated seedlings of gerbera jamesonii bolus |
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