CN107950397A - The method for tissue culture of American Red rocket crape myrtle - Google Patents
The method for tissue culture of American Red rocket crape myrtle Download PDFInfo
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- CN107950397A CN107950397A CN201711348392.0A CN201711348392A CN107950397A CN 107950397 A CN107950397 A CN 107950397A CN 201711348392 A CN201711348392 A CN 201711348392A CN 107950397 A CN107950397 A CN 107950397A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- Developmental Biology & Embryology (AREA)
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Abstract
The present invention proposes a kind of method for tissue culture of American Red rocket crape myrtle, comprises the following steps：1) it is inoculated with the sterilization of branch；2) inoculated and cultured：The explant of selection is cut into 1~2 centimetre of segment to be seeded on the inoculation medium configured, is cultivated 25~30 days；3) squamous subculture：Cultivated cutting in Multiple Buds inoculation subculture medium, in 23~27 DEG C of temperature, intensity of illumination 2200~2400lux, when daily illumination 10~12 is small, to 28~32 days, budlet base portion formed callus, and forms Multiple Buds, and the budlet that grows thickly reaches 1~2cm；4) culture of rootage：When seedling length is to 2~3 ㎝, it is transferred on root media and cultivates, for temperature control at 25~28 DEG C, intensity of illumination is 1100~1300lux, through 20~25 days when daily illumination 10~12 is small；5) hardening and transplanting.The method for tissue culture has broken the limitation of season and weather, and the breeding cycle shortens, and regeneration plant survival rate is improved, and survival rate can reach more than 95%.
Description
Technical field
The invention belongs to plant tissue culture technical field, and in particular to a kind of method for tissue culture of American Red rocket crape myrtle.
Background technology
Crape myrtle is Lythraceae (Lythraceae) Lagerstroemia defoliation small arbor or shrub, also known as crape myrtle, itch tree, full
Hall is red etc., and tree-like grace, blooms in the hot summer, and pattern is gorgeous, and florescence length, is the excellent ornamental plantation seeds in China.
American Red rocket crape myrtle is one of kind that pattern is most scarlet in crape myrtle, spends scarlet such as national flag, flowers are up to 24 English
Very little, flower amount is big, and pattern is gorgeous, the sea of flame when color lump that piece is planted is bloomed.Hua Qing hot day is scarlet, cloudy nice and cool day pattern
It is slightly light, there may be leukasmus, young leaves is micro- red, old leaf green slightly blush, and leaf bud is before flowering peony, becomes cherry when blooming
It is pink.First flower at the beginning of 6 months, can continuously reach the Frost's Descent, and the florescence is trimmed, and can repeatedly bloom.Wide adaptability, resistance are strong.It is adapted to
In isolated planting, group planting, cultivate to spend hedge, potted landscape and miniature gardening can be made.
The method for tissue culture of American Red rocket crape myrtle is common culturing and reproducing, but existing tissue cultures side
Method survival rate is relatively low, is easily subject to seasonal restrictions.
The content of the invention
The present invention proposes a kind of method for tissue culture of American Red rocket crape myrtle, the method for tissue culture broken season with
The limitation of weather, breeding cycle shorten, and regeneration plant survival rate is improved, and survival rate can reach more than 95%.
The technical proposal of the invention is realized in this way：
A kind of method for tissue culture of American Red rocket crape myrtle, comprises the following steps：
1) it is inoculated with the sterilization of branch：Choose American Red rocket crape myrtle and give birth to shoot then, carry out disinfection, after disinfection
Spray, which is seeded to, to lure on bud culture medium；
2) inoculated and cultured：The explant of selection is cut into 1~2 centimetre of segment and is seeded in the inoculation medium that has configured
On, cultivate 25~30 days；
3) squamous subculture：Cultivated cutting in Multiple Buds inoculation subculture medium, in 23~27 DEG C of temperature, illumination is strong
2200~2400lux is spent, when daily illumination 10~12 is small, to 28~32 days, budlet base portion formed callus, and is formed and grown thickly
Bud, the budlet that grows thickly reach 1~2cm；
4) culture of rootage：When seedling length is to 2~3 ㎝, it is transferred on root media and cultivates, temperature control is 25~28
DEG C, intensity of illumination is 1100~1300lux, through 20~25 days when daily illumination 10~12 is small；
5) hardening and transplanting：American Red rocket crape myrtle training tissue culture seedling, is first twisted in the greenhouse between 20~25 DEG C of temperature
Loose bottle cap is placed 2~4 days, then carries out hotbed transition cultivating, and transition hotbed is built in the vinyl house of conventional monomer, transplanting
During reduce to the greatest extent and hinder root, first spray carbendazim solution, then root carried out to carry out spraying treatment with nutrient solution, controls temperature
At 20~30 degree, transition cultivating 48~52 days, upper pot transplanting；
Wherein, the component of the inoculated and cultured culture medium is：ZW culture medium 2,4-D+0.3mg/L of+1.5mg/L NAA+
32~38g/L sucrose+6.5~7.5g/L carragheen+3.2~3.6g/L carbon dusts；The component of the subculture medium is：ZW is cultivated
6-BA+32 of base+0.5mg/L 2,4-D+0.4mg/L NAA+0.2mg/L~38g/L+6.5~7.5g/L of sucrose carragheens+
3.2~3.6g/L carbon dust+150~200g/L potatoes.
Preferably, the component of the root media is：ZW culture medium+0.8mg/L the NAA+ of 1/3 weight content
0.2mg/L IBA+0.25mg/L6-BA+0.3wt% activated carbon+22~28g/L sucrose.
Preferably, the pH value of the inoculation medium is 5.4~5.6, and the pH value of the subculture medium is 5.5~5.7,
The pH value of the root media is 5.5~5.7.
Preferably, the ZW culture mediums contain following component：1200mg/L ammonium nitrate+400mg/L potassium nitrate+450mg/L
Calcium sulfate+440mg/L calcium nitrate+76mg/L calcium chloride+260mg/L magnesium sulfate+170mg/L potassium dihydrogen phosphate+8.6mg/L sulfuric acid
Zinc+6.2mg/L boric acid+0.83mg/L potassium iodide+22.3mg/L manganese sulfate+0.025mg/L cobalt chloride+27.8mg/L ferrous sulfate
+ 37.3mg/L EDETATE SODIUM salt+100mg/L inositol+4.0mg/L glycine+1.2mg/L thiamine hydrochloride+2.5mg/L nicotinic acid+
1.2mg/L puridoxine hydrochloride+2.4mg/L D-VB5 calcium+2.0mg/L ascorbic acid, pH 5.6~5.8.
Preferably, the component of the nutrient solution is：0.1~0.12wt%NAA, 0.15~0.2wt% paclobutrazols, 0.03~
0.06wt% glycine, 0.2~0.3wt% vitamin Bs and 0.18~0.36wt% choline chlorides, surplus are water.
Preferably, the sterilization method of step 1) the American Red rocket crape myrtle shoot is：Shoot is soaked with bleaching powder
Bubble 5~after ten minutes, flowing water rinses 20~30 minutes, is soaked 2~3 minutes with 75% alcohol on superclean bench, after taking-up
With preprepared aseptic water washing 4~5 times, after draining the water.
Beneficial effects of the present invention：
The method of the present invention makes the breeding of American Red rocket crape myrtle more easy, has broken the limitation of season and weather, numerous
Cycle time is grown, regeneration plant survival rate is improved, and survival rate can reach more than 95%.
Embodiment
Embodiment 1
A kind of method for tissue culture of American Red rocket crape myrtle, comprises the following steps：
1) it is inoculated with the sterilization of branch：Choose American Red rocket crape myrtle and give birth to shoot then, by shoot bleaching powder
After five minutes, flowing water rinses 20 minutes for immersion, is soaked 2 minutes with 75% alcohol on superclean bench, with preparing in advance after taking-up
Good aseptic water washing 5 times, drains the water, the spray after disinfection, which is seeded to, to lure on bud culture medium；
2) inoculated and cultured：The explant of selection is cut into 1 centimetre of segment to be seeded on the inoculation medium configured, is trained
Support 25 days；
3) squamous subculture：Cultivated cutting in Multiple Buds inoculation subculture medium, in 23~27 DEG C of temperature, illumination is strong
2300lux is spent, when daily illumination 10 is small, to 30 days, budlet base portion formed callus, and forms Multiple Buds, and the budlet that grows thickly reaches
To 1~2cm；
4) culture of rootage：When seedling length is to 2~3 ㎝, it is transferred on root media and cultivates, temperature control is 25~28
DEG C, intensity of illumination 1100lux, through 23 days when daily illumination 11 is small；
5) hardening and transplanting：American Red rocket crape myrtle training tissue culture seedling, is first twisted in the greenhouse between 20~25 DEG C of temperature
Loose bottle cap is placed 3 days, then carries out hotbed transition cultivating, and transition hotbed is built in the vinyl house of conventional monomer, the mistake of transplanting
Reduced to the greatest extent in journey and hinder root, first spray carbendazim solution, then root is carried out to carry out spraying treatment with nutrient solution, control temperature exists
25 degree, transition cultivating 50 days, upper pot transplanting；
Wherein, the component of the inoculated and cultured culture medium is：ZW culture medium 2,4-D+0.3mg/L of+1.5mg/L NAA+
32g/L sucrose+6.5g/L carragheen+3.2g/L carbon dusts, the pH value of inoculation medium is 5.4~5.6.
The component of subculture medium is：2,4-D+0.4mg/LNAA+0.2mg/L6 of ZW culture medium+0.5mg/L-BA+
38g/L sucrose+7.5g/L carragheen+3.6g/L carbon dust+200g/L potatoes, the pH value of subculture medium is 5.5~5.7.
The component of root media is：ZW culture medium+0.8mg/LNAA+0.2mg/L the IBA+ of 1/3 weight content
0.25mg/L6-BA+0.3wt% activated carbon+26g/L sucrose, the pH value of root media is 5.5~5.7.
The component of nutrient solution is：0.1wt%NAA, 0.2wt% paclobutrazol, 0.03wt% glycine, 0.3wt% vitamin Bs
And 0.18wt% choline chlorides, surplus are water.
ZW culture mediums contain following component：1200mg/L ammonium nitrate+400mg/L potassium nitrate+450mg/L calcium sulfate+
440mg/L calcium nitrate+76mg/L calcium chloride+260mg/L magnesium sulfate+170mg/L potassium dihydrogen phosphate+8.6mg/L zinc sulfate+
6.2mg/L boric acid+0.83mg/L potassium iodide+22.3mg/L manganese sulfate+0.025mg/L cobalt chloride+27.8mg/L ferrous sulfate+
37.3mg/L EDETATE SODIUM salt+100mg/L inositol+4.0mg/L glycine+1.2mg/L thiamine hydrochloride+2.5mg/L nicotinic acid+
1.2mg/L puridoxine hydrochloride+2.4mg/L D-VB5 calcium+2.0mg/L ascorbic acid, pH 5.6~5.8.
Embodiment 2
A kind of method for tissue culture of American Red rocket crape myrtle, comprises the following steps：
1) it is inoculated with the sterilization of branch：Choose American Red rocket crape myrtle and give birth to shoot then, by shoot bleaching powder
After ten minutes, flowing water rinses 30 minutes for immersion, is soaked 3 minutes with 75% alcohol on superclean bench, with accurate in advance after taking-up
The aseptic water washing got ready 4 times, after draining the water, the spray after disinfection, which is seeded to, to lure on bud culture medium；
2) inoculated and cultured：The explant of selection is cut into 1~2 centimetre of segment and is seeded in the inoculation medium that has configured
On, cultivate 30 days；
3) squamous subculture：Cultivated cutting in Multiple Buds inoculation subculture medium, in 23~27 DEG C of temperature, illumination is strong
2200lux is spent, when daily illumination 12 is small, to 32 days, budlet base portion formed callus, and forms Multiple Buds, and the budlet that grows thickly reaches
To 1~2cm；
4) culture of rootage：When seedling length is to 2~3 ㎝, it is transferred on root media and cultivates, temperature control is 25~28
DEG C, intensity of illumination 1200lux, through 25 days when daily illumination 12 is small；
5) hardening and transplanting：American Red rocket crape myrtle training tissue culture seedling, is first twisted in the greenhouse between 20~25 DEG C of temperature
Loose bottle cap is placed 4 days, then carries out hotbed transition cultivating, and transition hotbed is built in the vinyl house of conventional monomer, the mistake of transplanting
Reduced to the greatest extent in journey and hinder root, first spray carbendazim solution, then root is carried out to carry out spraying treatment with nutrient solution, control temperature exists
30 degree, transition cultivating 48 days, upper pot transplanting；
Wherein, the component of inoculated and cultured culture medium is：ZW culture medium 2,4-D+0.3mg/L of+1.5mg/L NAA+38g/L
Sucrose+7.5g/L carragheen+3.6g/L carbon dusts, the pH value of inoculation medium is 5.4~5.6；
The component of subculture medium is：2,4-D+0.4mg/LNAA+0.2mg/L6 of ZW culture medium+0.5mg/L-BA+
32g/L sucrose+6.5g/L carragheen+3.2g/L carbon dust+150g/L potatoes, the pH value of subculture medium is 5.5~5.7.
The component of root media is：ZW culture medium+0.8mg/LNAA+0.2mg/L the IBA+ of 1/3 weight content
0.25mg/L6-BA+0.3wt% activated carbon+28g/L sucrose, the pH value of root media is 5.5~5.7.
The component of nutrient solution is：0.12wt%NAA, 0.15wt% paclobutrazol, 0.05wt% glycine, 0.2wt% dimension lifes
Plain B and 0.26wt% choline chlorides, surplus are water.
ZW culture mediums contain following component：1200mg/L ammonium nitrate+400mg/L potassium nitrate+450mg/L calcium sulfate+
440mg/L calcium nitrate+76mg/L calcium chloride+260mg/L magnesium sulfate+170mg/L potassium dihydrogen phosphate+8.6mg/L zinc sulfate+
6.2mg/L boric acid+0.83mg/L potassium iodide+22.3mg/L manganese sulfate+0.025mg/L cobalt chloride+27.8mg/L ferrous sulfate+
37.3mg/L EDETATE SODIUM salt+100mg/L inositol+4.0mg/L glycine+1.2mg/L thiamine hydrochloride+2.5mg/L nicotinic acid+
1.2mg/L puridoxine hydrochloride+2.4mg/L D-VB5 calcium+2.0mg/L ascorbic acid, pH 5.6~5.8.
Embodiment 3
A kind of method for tissue culture of American Red rocket crape myrtle, comprises the following steps：
1) it is inoculated with the sterilization of branch：Choose American Red rocket crape myrtle and give birth to shoot then, by shoot bleaching powder
After immersion 8 minutes, flowing water rinses 25 minutes, is soaked 3 minutes with 75% alcohol on superclean bench, with preparing in advance after taking-up
Good aseptic water washing 5 times, the spray after draining the water, which is seeded to, to lure on bud culture medium；
2) inoculated and cultured：The explant of selection is cut into 1~2 centimetre of segment and is seeded in the inoculation medium that has configured
On, cultivate 30 days；
3) squamous subculture：Cultivated cutting in Multiple Buds inoculation subculture medium, in 23~27 DEG C of temperature, illumination is strong
2400lux is spent, when daily illumination 10 is small, to 28 days, budlet base portion formed callus, and forms Multiple Buds, and the budlet that grows thickly reaches
To 1~2cm；
4) culture of rootage：When seedling length is to 2~3 ㎝, it is transferred on root media and cultivates, temperature control is 25~28
DEG C, intensity of illumination 1300lux, through 20 days when daily illumination 10 is small；
5) hardening and transplanting：American Red rocket crape myrtle training tissue culture seedling, is first twisted in the greenhouse between 20~25 DEG C of temperature
Loose bottle cap is placed 4 days, then carries out hotbed transition cultivating, and transition hotbed is built in the vinyl house of conventional monomer, the mistake of transplanting
Reduced to the greatest extent in journey and hinder root, first spray carbendazim solution, then root is carried out to carry out spraying treatment with nutrient solution, control temperature exists
20 degree, transition cultivating 52 days, upper pot transplanting；
Wherein, the component of inoculated and cultured culture medium is：ZW culture medium 2,4-D+0.3mg/L of+1.5mg/L NAA+36g/L
Sucrose+7g/L carragheen+3.5g/L carbon dusts, pH value are 5.4~5.6.
The component of subculture medium is：2,4-D+0.4mg/LNAA+0.2mg/L6 of ZW culture medium+0.5mg/L-BA+
36g/L sucrose+7g/L carragheen+3.4g/L carbon dust+180g/L potatoes, pH value are 5.5~5.7.
The component of root media is：ZW culture medium+0.8mg/LNAA+0.2mg/L the IBA+ of 1/3 weight content
0.25mg/L6-BA+0.3wt% activated carbon+25g/L sucrose, pH value are 5.5~5.7.
The component of nutrient solution is：0.12wt%NAA, 0.18wt% paclobutrazol, 0.06wt% glycine, 0.2wt% dimension lifes
Plain B and 0.36wt% choline chlorides, surplus are water.
ZW culture mediums contain following component：1200mg/L ammonium nitrate+400mg/L potassium nitrate+450mg/L calcium sulfate+
440mg/L calcium nitrate+76mg/L calcium chloride+260mg/L magnesium sulfate+170mg/L potassium dihydrogen phosphate+8.6mg/L zinc sulfate+
6.2mg/L boric acid+0.83mg/L potassium iodide+22.3mg/L manganese sulfate+0.025mg/L cobalt chloride+27.8mg/L ferrous sulfate+
37.3mg/L EDETATE SODIUM salt+100mg/L inositol+4.0mg/L glycine+1.2mg/L thiamine hydrochloride+2.5mg/L nicotinic acid+
1.2mg/L puridoxine hydrochloride+2.4mg/L D-VB5 calcium+2.0mg/L ascorbic acid, pH 5.6~5.8.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
With within principle, any modification, equivalent replacement, improvement and so on, should all be included in the protection scope of the present invention god.
Claims (6)
1. a kind of method for tissue culture of American Red rocket crape myrtle, it is characterised in that comprise the following steps：
1) it is inoculated with the sterilization of branch：Choose American Red rocket crape myrtle and give birth to shoot then, carry out disinfection, the spray after disinfection
It is seeded to and lures on bud culture medium；
2) inoculated and cultured：The explant of selection is cut into 1~2 centimetre of segment to be seeded on the inoculation medium configured, is trained
Support 25~30 days；
3) squamous subculture：Cultivated cutting in Multiple Buds inoculation subculture medium, in 23~27 DEG C of temperature, intensity of illumination
When 2200~2400lux, daily illumination 10~12 are small, to 28~32 days, budlet base portion formed callus, and is formed and grown thickly
Bud, the budlet that grows thickly reach 1~2cm；
4) culture of rootage：When seedling length is to 2~3 ㎝, is transferred on root media and cultivates, temperature control at 25~28 DEG C,
Intensity of illumination is 1100~1300lux, through 20~25 days when daily illumination 10~12 is small；
5) hardening and transplanting：American Red rocket crape myrtle training tissue culture seedling, first unscrews bottle in the greenhouse between 20~25 DEG C of temperature
Lid is placed 2~4 days, then carries out hotbed transition cultivating, and transition hotbed is built in the vinyl house of conventional monomer, the process of transplanting
In reduce to the greatest extent and hinder root, first spray carbendazim solution, then root carried out to carry out spraying treatment with nutrient solution, control temperature is 20
~30 degree, transition cultivating 48~52 days, upper pot transplanting；
Wherein, the component of the inoculated and cultured culture medium is：ZW culture medium 2,4-D+0.3mg/L of+1.5mg/L NAA+32~
38g/L sucrose+6.5~7.5g/L carragheen+3.2~3.6g/L carbon dusts；The component of the subculture medium is：ZW culture mediums+
6-BA+32 of 0.5mg/L 2,4-D+0.4mg/L NAA+0.2mg/L~38g/L+6.5~7.5g/L of sucrose carragheens+3.2
~3.6g/L carbon dust+150~200g/L potatoes.
2. the method for tissue culture of American Red rocket crape myrtle according to claim 1, it is characterised in that the culture of rootage
The component of base is：ZW culture medium+0.8mg/L NAA+0.2mg/L IBA+0.25mg/L6-BA+ of 1/3 weight content
0.3wt% activated carbon+22~28g/L sucrose.
3. the method for tissue culture of American Red rocket crape myrtle according to claim 2, it is characterised in that the inoculated and cultured
The pH value of base is 5.4~5.6, and the pH value of the subculture medium is 5.5~5.7, the pH value of the root media for 5.5~
5.7。
4. the method for tissue culture of American Red rocket crape myrtle according to claim 1 or 2, it is characterised in that the ZW trainings
Foster base contains following component：1200mg/L ammonium nitrate+400mg/L potassium nitrate+450mg/L calcium sulfate+440mg/L calcium nitrate+
76mg/L calcium chloride+260mg/L magnesium sulfate+170mg/L potassium dihydrogen phosphate+8.6mg/L zinc sulfate+6.2mg/L boric acid+
0.83mg/L potassium iodide+22.3mg/L manganese sulfate+0.025mg/L cobalt chloride+27.8mg/L ferrous sulfate+37.3mg/L EDTA
Sodium salt+100mg/L inositol+4.0mg/L glycine+1.2mg/L thiamine hydrochloride+2.5mg/L nicotinic acid+1.2mg/L hydrochloric acid pyrroles are trembled
Alcohol+2.4mg/L D-VB5 calcium+2.0mg/L ascorbic acid, pH 5.6~5.8.
5. the method for tissue culture of American Red rocket crape myrtle according to claim 1, it is characterised in that the nutrient solution
Component is：0.1~0.12wt%NAA, 0.15~0.2wt% paclobutrazols, 0.03~0.06wt% glycine, 0.2~0.3wt%
Vitamin B and 0.18~0.36wt% choline chlorides, surplus are water.
6. the method for tissue culture of American Red rocket crape myrtle according to claim 1, it is characterised in that the step 1) is beautiful
The sterilization method of the flourishing arrow crape myrtle shoot of state is：By shoot with bleaching powder immersion 5~after ten minutes, flowing water rinse 20~30
Minute, soaked 2~3 minutes with 75% alcohol on superclean bench, with preprepared aseptic water washing 4~5 after taking-up
Time, after draining the water.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109169289A (en) * | 2018-11-06 | 2019-01-11 | 湖南省林业科学院 | A kind of crape myrtle tissue culture proliferation seedling rooting medicament and method |
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| CN104115661A (en) * | 2014-07-21 | 2014-10-29 | 江苏绿苑园林建设有限公司 | American lagerstroemia indica red rocket branch cutting propagation method |
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| CN104115661A (en) * | 2014-07-21 | 2014-10-29 | 江苏绿苑园林建设有限公司 | American lagerstroemia indica red rocket branch cutting propagation method |
| CN104126506A (en) * | 2014-07-21 | 2014-11-05 | 江苏绿苑园林建设有限公司 | Tissue culture method of America Lagerstroemia indica Red Rocket |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109169289A (en) * | 2018-11-06 | 2019-01-11 | 湖南省林业科学院 | A kind of crape myrtle tissue culture proliferation seedling rooting medicament and method |
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