CN104304009B - Siberian wildrye young ear isolated culture regeneration plant method - Google Patents
Siberian wildrye young ear isolated culture regeneration plant method Download PDFInfo
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Abstract
The invention mainly relates to a siberian wildrye young ear isolated culture plant regeneration technology, which belongs to the technical field of biology. The method is characterized by comprising the following steps: (1) acquiring an explant; (2) inducing a callus; (3) inoculating the induced callus into a subculture medium to be cultured; (4) transferring the siberian wildrye callus of the step (3) into a differential medium to be cultured; (5) transferring the Siberian wildrye callus into a rooting seedling culture medium to be cultured when the Siberian wildrye callus has green shoots; and (6) removing a sealing film on a triangular flask when more than 5 roots grow up, adding 5mL of distilled water, exercising the seedlings for 3 days, taking out the small plant from the triangular flask, washing the culture medium on the roots, finally transplanting the small plant into a flowerpot with the turfy soil and vermiculite in the ratio of 1:1, and covering the seedlings with a plastic film to maintain the moisture in the first three days. By adopting the method, a research platform is provided for carrying out the genetic transformation and character improvement.
Description
Technical field
The present invention relates to one kind carries out tissue culture with siberian wildrye children's fringe for explant, the method obtaining regeneration plant, belong to
Pass through the plant regeneration field of tissue culture technique in agricultural cultivation.
Background technology
Siberian wildrye (elymus sibiricus) also known as vertical fringe Herba Hordei Vulgaris, Siberia lyme grass, are grass family lyme grass
Belong to perennial herb and dredge clump type plant, widely distributed in the area such as the northeast of China, North China, northwest and Qinghai-Tibet Platean, it is grassy marshland
Sociales in grassland and hygropium and constructive species (Zhou Yonghong etc., 1999；Chen Mojun and Jia Shenxiu, 2002).Because of its tiller
Power is strong, and cold-resistant, drought resistance is good, and grass yield is high, is the good forage extensively cultivated.Additionally, siberian wildrye can set up list
One artificial mowing grassland and grazing ground it is also possible to set up the artificial pasture of high-quality, high yield with other dogstail and leguminous forage mixed seeding；
Plant Leaf amount is enriched, and nutrition branch is many, and concentration is compared in spike opening, and setting percentage is high, and seed production and grass yield are in Second Year
Reach summit (king's Bhide, 1986；Ma Zong Renhe Guo Bo, 1991；Wu Qin, 1992；Wu Baoguo, 2003).Xiao Bo etc. (2010)
By studying to the effect of fostering and apply fertilizer of siberian wildrye, while to find it be that people bring economy, ecological benefits, top layer can be improved
The physicochemical property of soil, and play an important role in the scientific cultivation of wasteland soil.
Tissue culture refers to aseptically using synthetic medium, plant tissue be cultivated, and is that biological engineering grinds
The basis studied carefully, is by genetic transformation and the basic link of plant improvement research.Li Daxu etc. (2006) establishes Sichuan Cordyceps two
The whole plant regenerating system of siberian wildrye mature seed, hypocotyls and spire；Lee etc. (2012) utilizes Siberia siberian wildrye to become
Ripe Seed inducement wound healing and be differentiated to form regeneration plant.But up to the present, organized with siberian wildrye children's fringe for explant
The research of culture is also rarely reported, and hence sets up necessary with siberian wildrye children's Tissue Culture Regeneration System as explant for the fringe,
Also lay a good foundation for transgenic research from now on simultaneously.
Content of the invention
The technical problem to be solved in the present invention and the technical assignment proposing are to overcome existing siberian wildrye Techniques of in Vitro Culture to produce
The defect such as raw pollution rate is high, Callus induction rate is low and reproducibility is poor, provides a kind of siberian wildrye children's fringe isolated culture plant strain regeneration
Method, as explant material, regulates and controls the osmotic pressure of exogenous plant hormones and culture medium, obtains using by the young fringe from siberian wildrye
Obtain a high proportion of pellet embryogenic callus, set up the technical system of plant regeneration, for carrying out genetic transformation and character improvement
Research platform is provided.
The present invention employs the following technical solutions: a kind of method of siberian wildrye children's fringe isolated culture regeneration plant, it is mainly special
Point is to comprise the following steps:
1) acquisition of explant: take the siberian wildrye children's fringe being in Pistil And Stamen idiophase, length for 3.0-10cm, clip plant
The first half, aseptically dips its stalk of 70% ethanol and sheath with cotton balls after removing the blade of outer layer, with disappearing
The tweezers of poison strip young fringe, are cut into the segment of 2-3mm；
2) callus induction: explant is inoculated on calli induction media, 10 young fringe segments inoculated by every ware, put
Enter culturing room's culture, culture room temperature is 24-26 DEG C, light application time 16h/d, interlunation 8h/d, and incubation time is 1-2
Month；
3) calluss of induction are inoculated in subculture medium and are cultivated, culture room temperature is 24-26 DEG C, illumination
Time 16h/d, interlunation 8h/d, cultivated days are 20-30d；
4) by step 3) siberian wildrye calluss be transferred in division culture medium culture, culture room temperature is 24-26 DEG C,
Light application time 16h/d, interlunation 8h/d, cultivated days are 20-30d；
5) treat step 4) siberian wildrye calluss have green bud to grow, transfer them to and trained in seedling culture medium of taking root
Support, culture room temperature is 24-26 DEG C, light application time 16h/d, interlunation 8h/d；
6) treated that more than 5 healthy and strong roots grow, the sealed membrane on triangular flask has been removed, is simultaneously introduced the distilled water of 5ml, refining
Then plantlet is taken out from triangular flask by Seedling 3d, the culture medium of clean root, finally transplants in being mixed with turfy soil and Vermiculitum ratio
In the flowerpot for 1:1 for the example, initial 3d hides upper plastic sheeting to keep humidity to seedling.
The method of described siberian wildrye children's fringe isolated culture regeneration plant, described callus inducing medium is: nitre
Sour potassium 1900mg/l, ammonium nitrate 1650mg/l, bitter salt 370mg/l, CALCIUM CHLORIDE DIHYDRATE 440mg/l, biphosphate
Potassium 170mg/l, four anhydrous manganese 22.3mg/l, Zinc vitriol 8.6mg/l, boric acid 6.2mg/l, potassium iodide 0.83mg/
L, two molybdic acid hydrate sodium 0.25mg/l, copper sulfate pentahydrate 0.025mg/l, cobalt chloride hexahydrate 0.025mg/l, inositol 100mg/
L, glycine 2.0mg/l, pyridoxine hydrochloride 0.5mg/l, thiamine hydrochloride 0.1mg/l, nicotinic acid 0.5mg/l, ethylenediaminetetraacetic acid
Disodium 37.3mg/l, green vitriol 27.8mg/l, sucrose 30g/l, agar 7g/l, 2,4- dichlorphenoxyacetic acid 2.5mg/
l.
The method of described siberian wildrye children's fringe isolated culture regeneration plant, described subculture medium is: potassium nitrate
1900mg/l, ammonium nitrate 1650mg/l, bitter salt 370mg/l, CALCIUM CHLORIDE DIHYDRATE 440mg/l, potassium dihydrogen phosphate
170mg/l, four anhydrous manganese 22.3mg/l, Zinc vitriol 8.6mg/l, boric acid 6.2mg/l, potassium iodide 0.83mg/l,
Two molybdic acid hydrate sodium 0.25mg/l, copper sulfate pentahydrate 0.025mg/l, cobalt chloride hexahydrate 0.025mg/l, inositol 100mg/l,
Glycine 2.0mg/l, pyridoxine hydrochloride 0.5mg/l, thiamine hydrochloride 0.1mg/l, nicotinic acid 0.5mg/l, ethylenediaminetetraacetic acid two
Sodium 37.3mg/l, green vitriol 27.8mg/l, sucrose 30g/l, agar 7g/l, 2,4- dichlorphenoxyacetic acid 1.5mg/l.
The method of described siberian wildrye children's fringe isolated culture regeneration plant, described division culture medium is: potassium nitrate
1900mg/l, ammonium nitrate 1650mg/l, bitter salt 370mg/l, CALCIUM CHLORIDE DIHYDRATE 440mg/l, potassium dihydrogen phosphate
170mg/l, four anhydrous manganese 22.3mg/l, Zinc vitriol 8.6mg/l, boric acid 6.2mg/l, potassium iodide 0.83mg/l,
Two molybdic acid hydrate sodium 0.25mg/l, copper sulfate pentahydrate 0.025mg/l, cobalt chloride hexahydrate 0.025mg/l, inositol 100mg/l,
Glycine 2.0mg/l, pyridoxine hydrochloride 0.5mg/l, thiamine hydrochloride 0.1mg/l, nicotinic acid 0.5mg/l, ethylenediaminetetraacetic acid two
Sodium 37.3mg/l, green vitriol 27.8mg/l, sucrose 30g/l, agar 7g/l, kinetins 1.0mg/l and 6- ammonia benzyl
Adenine 1.0mg/l.
The method of described siberian wildrye children's fringe isolated culture regeneration plant, described seedling culture medium of taking root is: 1/2ms training
Foster base is that potassium nitrate 950mg/l, ammonium nitrate 825mg/l, potassium dihydrogen phosphate 85mg/l, bitter salt 185mg/l, two are hydrated
Calcium chloride 220mg/l, four anhydrous manganese 22.3mg/l, Zinc vitriol 8.6mg/l, boric acid 6.2mg/l, potassium iodide
0.83mg/l, two molybdic acid hydrate sodium 0.25mg/l, copper sulfate pentahydrate 0.025mg/l, cobalt chloride hexahydrate 0.025mg/l, flesh
Alcohol 100mg/l, glycine 2.0mg/l, pyridoxine hydrochloride 0.5mg/l, thiamine hydrochloride 0.1mg/l, nicotinic acid 0.5mg/l, second two
Amine tetraacethyl disodium 37.3mg/l, green vitriol 27.8mg/l, sucrose 30g/l, agar 7g/l.
The present invention compared with prior art has the advantage that and good effect:
Being explant from be in Pistil And Stamen breaking up period, the length about young fringe of 3.0-10cm, carrying out in vitro tissue again
Raw culture.In regulation and control callus inducing medium, the proportioning of exogenous hormone, improves the induction of siberian wildrye embryo callus
Rate.Reduce calluss subculture medium in hormone 2, the concentration of 4- dichlorphenoxyacetic acid to 1.5mg/l, to improve calluss
Quality and cells,primordial incidence rate.Siberian wildrye children's fringe isolated culture plant strain regeneration method system that the present invention sets up, for carrying out
Genetic transformation and character improvement provide research platform.
Brief description
Fig. 1: young fringe callus induction；
Fig. 2: calluss Differentiation From Calli；
Fig. 3: the seedling taking root；
Fig. 4: transplant the regeneration plant to flowerpot.
Specific embodiment
With reference to embodiments the principle and feature of the present invention is described, example is served only for explaining the present invention,
It is not intended to limit the scope of the present invention.
Embodiment 1: a kind of method of siberian wildrye children's fringe isolated culture regeneration plant, it is mainly characterized by and walks including following
Rapid:
1) acquisition of explant: take the siberian wildrye children's fringe being in Pistil And Stamen idiophase, length for 3.0-10cm, clip plant
The first half, aseptically dips its stalk of 70% ethanol and sheath with cotton balls after removing the blade of outer layer, with disappearing
The tweezers of poison strip young fringe, are cut into the segment of 2-3mm；
2) callus induction: explant is inoculated on calli induction media, every ware inoculation children's fringe segment, puts into training
The culture of foster room, culture room temperature is 24-26 DEG C, light application time 16h/d, interlunation 8h/d, and incubation time is 1-2 month；
3) calluss of induction are inoculated in subculture medium and are cultivated, culture room temperature is 24-26 DEG C, illumination
Time 16h/d, interlunation 8h/d, cultivated days are 20-30d；
4) by step 3) siberian wildrye calluss be transferred in division culture medium culture, culture room temperature is 24-26 DEG C,
Light application time 16h/d, interlunation 8h/d, cultivated days are 20-30d；
5) treat step 4) siberian wildrye calluss have green bud to grow, transfer them to and trained in seedling culture medium of taking root
Support, culture room temperature is 24-26 DEG C, light application time 16h/d, interlunation 8h/d；
6) treated that more than 5 healthy and strong roots grow, the sealed membrane on triangular flask has been removed, is simultaneously introduced the distilled water of 5ml, refining
Then plantlet is taken out from triangular flask by Seedling 3d, the culture medium of clean root, finally transplants in being mixed with turfy soil and Vermiculitum ratio
In the flowerpot for 1:1 for the example, initial 3d hides upper plastic sheeting to keep humidity to seedling.
The method of described siberian wildrye children's fringe isolated culture regeneration plant, described callus inducing medium is: nitre
Sour potassium 1900mg/l, ammonium nitrate 1650mg/l, bitter salt 370mg/l, CALCIUM CHLORIDE DIHYDRATE 440mg/l, biphosphate
Potassium 170mg/l, four anhydrous manganese 22.3mg/l, Zinc vitriol 8.6mg/l, boric acid 6.2mg/l, potassium iodide 0.83mg/
L, two molybdic acid hydrate sodium 0.25mg/l, copper sulfate pentahydrate 0.025mg/l, cobalt chloride hexahydrate 0.025mg/l, inositol 100mg/
L, glycine 2.0mg/l, pyridoxine hydrochloride 0.5mg/l, thiamine hydrochloride 0.1mg/l, nicotinic acid 0.5mg/l, ethylenediaminetetraacetic acid
Disodium 37.3mg/l, green vitriol 27.8mg/l, sucrose 30g/l, agar 7g/l, 2,4- dichlorphenoxyacetic acid 2.5mg/
l.
The method of described siberian wildrye children's fringe isolated culture regeneration plant, described subculture medium is: potassium nitrate
1900mg/l, ammonium nitrate 1650mg/l, bitter salt 370mg/l, CALCIUM CHLORIDE DIHYDRATE 440mg/l, potassium dihydrogen phosphate
170mg/l, four anhydrous manganese 22.3mg/l, Zinc vitriol 8.6mg/l, boric acid 6.2mg/l, potassium iodide 0.83mg/l,
Two molybdic acid hydrate sodium 0.25mg/l, copper sulfate pentahydrate 0.025mg/l, cobalt chloride hexahydrate 0.025mg/l, inositol 100mg/l,
Glycine 2.0mg/l, pyridoxine hydrochloride 0.5mg/l, thiamine hydrochloride 0.1mg/l, nicotinic acid 0.5mg/l, ethylenediaminetetraacetic acid two
Sodium 37.3mg/l, green vitriol 27.8mg/l, sucrose 30g/l, agar 7g/l, 2,4- dichlorphenoxyacetic acid 1.5mg/l.
The method of described siberian wildrye children's fringe isolated culture regeneration plant, described division culture medium is: potassium nitrate
1900mg/l, ammonium nitrate 1650mg/l, bitter salt 370mg/l, CALCIUM CHLORIDE DIHYDRATE 440mg/l, potassium dihydrogen phosphate
170mg/l, four anhydrous manganese 22.3mg/l, Zinc vitriol 8.6mg/l, boric acid 6.2mg/l, potassium iodide 0.83mg/l,
Two molybdic acid hydrate sodium 0.25mg/l, copper sulfate pentahydrate 0.025mg/l, cobalt chloride hexahydrate 0.025mg/l, inositol 100mg/l,
Glycine 2.0mg/l, pyridoxine hydrochloride 0.5mg/l, thiamine hydrochloride 0.1mg/l, nicotinic acid 0.5mg/l, ethylenediaminetetraacetic acid two
Sodium 37.3mg/l, green vitriol 27.8mg/l, sucrose 30g/l, agar 7g/l, kinetins 1.0mg/l and 6- ammonia benzyl
Adenine 1.0mg/l.
The method of described siberian wildrye children's fringe isolated culture regeneration plant, described seedling culture medium of taking root is: 1/2ms training
Foster base is that potassium nitrate 950mg/l, ammonium nitrate 825mg/l, potassium dihydrogen phosphate 85mg/l, bitter salt 185mg/l, two are hydrated
Calcium chloride 220mg/l, four anhydrous manganese 22.3mg/l, Zinc vitriol 8.6mg/l, boric acid 6.2mg/l, potassium iodide
0.83mg/l, two molybdic acid hydrate sodium 0.25mg/l, copper sulfate pentahydrate 0.025mg/l, cobalt chloride hexahydrate 0.025mg/l, flesh
Alcohol 100mg/l, glycine 2.0mg/l, pyridoxine hydrochloride 0.5mg/l, thiamine hydrochloride 0.1mg/l, nicotinic acid 0.5mg/l, second two
Amine tetraacethyl disodium 37.3mg/l, green vitriol 27.8mg/l, sucrose 30g/l, agar 7g/l.
Verification experimental verification example: a kind of method of siberian wildrye children's fringe isolated culture regeneration plant, siberian wildrye picks up from Gansu Province Luqu
County's wild germplasm, its tissue culture procedures is:
1) acquisition of explant: take and be in the Pistil And Stamen idiophase, length is about the young fringe of 3.0-10cm, clip siberian wildrye is planted
The strain first half, aseptically dips its stalk of 70% ethanol and sheath with cotton balls after removing the blade of outer layer, with disappearing
The tweezers crossing poison carefully strip young fringe, are cut into the segment of 2-3mm.
2) callus induction: explant is inoculated on calli induction media, 10 young fringe segments inoculated by every ware, put
Enter culturing room's culture.
3) callus inducing medium formula: ms minimal medium adds 2,4- dichlorphenoxyacetic acid 2.5mg/l, culture
Room temperature is 24-26 DEG C, light application time 16h/d, interlunation 8h/d.
4) calluss of induction are inoculated in subculture medium and are cultivated.
5) successive transfer culture based formulas: ms minimal medium adds 2,4- dichlorphenoxyacetic acid 1.5mg/l, sucrose 30g/l, fine jade
Fat 7.0g/l, culture room temperature is 24-26 DEG C, light application time 16h/d, interlunation 8h/d.
6) cultivate 20-30d in subculture medium, siberian wildrye calluss are transferred to culture in division culture medium.
7) differentiation culture based formulas: ms minimal medium adds kinetins 1.0mg/l and 6- ammonia benzyladenine 1.0mg/
L, sucrose 30g/l, agar 7.0g/l, culture room temperature is 24-26 DEG C, light application time 16h/d, interlunation 8h/d.
8) in division culture medium, the time of culture is 20d.
9) treat that calluss have green bud to grow, transfer them to and cultivated in seedling culture medium of taking root.
10) seedling of taking root culture medium prescription: 1/2ms (ms culture medium a great number of elements halves, other constant), cultivates room temperature
For 24-26 DEG C, light application time 16h/d, interlunation 8h/d.
11) treated that more than 5 healthy and strong roots grow, the sealed membrane on triangular flask removed, is simultaneously introduced the distilled water of 5ml,
Then plantlet is taken out from triangular flask by seedling exercising 3d, the culture medium of clean root, finally transplants in being mixed with turfy soil and Vermiculitum
Ratio is in the flowerpot of 1:1, and initial 3d hides upper plastic sheeting to keep humidity to seedling.
Claims (1)
1. a kind of method of siberian wildrye children's fringe isolated culture regeneration plant is it is characterised in that comprise the following steps:
1) acquisition of explant: take the siberian wildrye children's fringe being in Pistil And Stamen idiophase, length for 3.0-10 cm, on clip plant
Half portion, aseptically dips its stalk of 70% ethanol and sheath with cotton balls after removing the blade of outer layer, with sterilized
Tweezers strip young fringe, are cut into the segment of 2-3 mm；
2) callus induction: explant is inoculated on calli induction media, 10 young fringe segments inoculated by every ware, put into training
The culture of foster room, culture room temperature is 24-26 DEG C, light application time 16h/d, interlunation 8 h/d, and incubation time is 1-2 month；
3) calluss of induction are inoculated in subculture medium and are cultivated, culture room temperature is 24-26 DEG C, during illumination
Between 16 h/d, interlunation 8 h/d, cultivated days be 20-30 d；
4) the siberian wildrye calluss of step 3) are transferred to culture in division culture medium, culture room temperature is 24-26 DEG C, light
According to time 16 h/d, interlunation 8 h/d, cultivated days are 20-30 d；
5) treat that the siberian wildrye calluss of step 4) have green bud to grow, transfer them to and cultivated in seedling culture medium of taking root,
Culture room temperature is 24-26 DEG C, light application time 16 h/d, interlunation 8 h/d；
6) treated that more than 5 healthy and strong roots grow, the sealed membrane on triangular flask has been removed, is simultaneously introduced the distilled water of 5 ml, seedling exercising 3
Then plantlet is taken out from triangular flask by d, the culture medium of clean root, finally transplants in being mixed with turfy soil and Vermiculitum ratio
For, in the flowerpot of 1:1, initial 3 d hide upper plastic sheeting to keep humidity to seedling；
Described callus inducing medium is: potassium nitrate 1900 mg/l, ammonium nitrate 1650 mg/l, bitter salt
370 mg/l, CALCIUM CHLORIDE DIHYDRATE 440 mg/l, potassium dihydrogen phosphate 170 mg/l, four anhydrous manganese 22.3 mg/l, seven are hydrated
Zinc sulfate 8.6 mg/l, boric acid 6.2 mg/l, potassium iodide 0.83 mg/l, two molybdic acid hydrate sodium 0.25 mg/l, five hydrated sulfuric acid
Copper 0.025 mg/l, cobalt chloride hexahydrate 0.025 mg/l, inositol 100 mg/l, glycine 2.0 mg/l, pyridoxine hydrochloride 0.5
Mg/l, thiamine hydrochloride 0.1 mg/l, nicotinic acid 0.5 mg/l, disodiumedetate 37.3 mg/l, green vitriol
27.8 mg/l, sucrose 30 g/l, agar 7 g/l, 2,4- dichlorphenoxyacetic acid 2.5 mg/l；
Described subculture medium is: potassium nitrate 1900 mg/l, ammonium nitrate 1650 mg/l, bitter salt 370 mg/l,
CALCIUM CHLORIDE DIHYDRATE 440 mg/l, potassium dihydrogen phosphate 170 mg/l, four anhydrous manganese 22.3 mg/l, Zinc vitriol 8.6
Mg/l, boric acid 6.2 mg/l, potassium iodide 0.83 mg/l, two molybdic acid hydrate sodium 0.25 mg/l, copper sulfate pentahydrate 0.025 mg/
L, cobalt chloride hexahydrate 0.025 mg/l, inositol 100 mg/l, glycine 2.0 mg/l, pyridoxine hydrochloride 0.5 mg/l, hydrochloric acid
Thiamine 0.1 mg/l, nicotinic acid 0.5 mg/l, disodiumedetate 37.3 mg/l, green vitriol 27.8 mg/
L, sucrose 30 g/l, agar 7.0 g/l, 2,4- dichlorphenoxyacetic acid 1.5 mg/l；
Described division culture medium is: potassium nitrate 1900 mg/l, ammonium nitrate 1650 mg/l, bitter salt 370 mg/l,
CALCIUM CHLORIDE DIHYDRATE 440 mg/l, potassium dihydrogen phosphate 170 mg/l, four anhydrous manganese 22.3 mg/l, Zinc vitriol 8.6
Mg/l, boric acid 6.2 mg/l, potassium iodide 0.83 mg/l, two molybdic acid hydrate sodium 0.25 mg/l, copper sulfate pentahydrate 0.025 mg/
L, cobalt chloride hexahydrate 0.025 mg/l, inositol 100 mg/l, glycine 2.0 mg/l, pyridoxine hydrochloride 0.5 mg/l, hydrochloric acid
Thiamine 0.1 mg/l, nicotinic acid 0.5 mg/l, disodiumedetate 37.3 mg/l, green vitriol 27.8 mg/
L, sucrose 30g/l, agar 7 g/l, kinetins 1.0 mg/l and 6- ammonia benzyladenine 1.0 mg/l；
Described seedling culture medium of taking root is 1/2 ms culture medium, and its concrete component is, potassium nitrate 950 mg/l, ammonium nitrate 825
Mg/l, potassium dihydrogen phosphate 85 mg/l, bitter salt 185 mg/l, CALCIUM CHLORIDE DIHYDRATE 220 mg/l, four anhydrous manganeses
22.3 mg/l, Zinc vitriol 8.6 mg/l, boric acid 6.2 mg/l, potassium iodide 0.83 mg/l, two molybdic acid hydrate sodium 0.25
Mg/l, copper sulfate pentahydrate 0.025 mg/l, cobalt chloride hexahydrate 0.025 mg/l, inositol 100 mg/l, glycine 2.0 mg/
L, pyridoxine hydrochloride 0.5 mg/l, thiamine hydrochloride 0.1 mg/l, nicotinic acid 0.5 mg/l, disodiumedetate 37.3 mg/
L, green vitriol 27.8 mg/l, sucrose 30 g/l, agar 7 g/l.
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| CN115885850A (en) * | 2022-11-30 | 2023-04-04 | 中国科学院西北高原生物研究所 | Tissue culture medium for regeneration of old mango wheat and tissue culture method for regeneration of mature embryo of old mango wheat |
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