CN104604735B - A kind of tissue culture and rapid propagation method of U.S.'s red autumnal leaves Lagerstroemia indica L. - Google Patents
A kind of tissue culture and rapid propagation method of U.S.'s red autumnal leaves Lagerstroemia indica L. Download PDFInfo
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- CN104604735B CN104604735B CN201510106620.8A CN201510106620A CN104604735B CN 104604735 B CN104604735 B CN 104604735B CN 201510106620 A CN201510106620 A CN 201510106620A CN 104604735 B CN104604735 B CN 104604735B
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Abstract
The invention discloses a kind of tissue culture and rapid propagation method of U.S.'s red autumnal leaves Lagerstroemia indica L., belongs to the technical field of tissue culture of plant.The present invention by carrying out isolated culture to U.S.'s red autumnal leaves Lagerstroemia indica L. semi-lignified branch, sterilize, initial culture, enrichment culture, root culture by the selection including explant, five stages of acclimatization and transplantses.The present invention has breeding coefficient high, and cultivation period is short, and the advantages of not being subject to seasonal restrictions, inductivity is up to 88%；, up to 5～6, the elongation growth of propagation bud is fast for growth coefficient, and culture 25d buds are up to 3～4cm；Up to 100%, mean elements is 4.54 pieces/plant to rooting rate；Tissue culture is taken root, and Seedling is healthy and strong, transplanting survival rate up to more than 95%；Anniversary scale fast seedling growing can be carried out in production application, produced the U.S. red autumnal leaves Lagerstroemia indica L. seedling of stalwartness, neat and consistent, had a extensive future.
Description
Technical field
The invention belongs to the technical field of tissue culture of plant, more particularly to a kind of tissue-culturing rapid propagation side of U.S.'s red autumnal leaves Lagerstroemia indica L.
Method.
Background technology
Lagerstroemia indica L. is Lythraceae (Lythraceae) Lagerstroemia defoliation small arbor or shrub, also known as Lagerstroemia indica L., itch tree, full
Hall is red etc., and tree-like grace was bloomed in the hot summer, and pattern is gorgeous, and the florescence is long, was the excellent ornamental plantation seeds of China.U.S.'s red autumnal leaves
Lagerstroemia indica L. (Lagerstroemia indica ' Pink Velour ') is the purple that was introduced from the U.S. in 2003 by forest-science academy of Hunan Province
The new quality product kind of common vetch, its young leaves and tender tip are claret, with good sight leaf effect；Flower rediance, inflorescence length up to 70cm,
Dense and graceful, with high ornamental value.The first tenday period of a month June at florescence to by the end of October, up to 5 months.Its strong adaptability, drought-enduring,
It is resistant to -23 DEG C of low temperature.The lace that is used in afforestation, flowering shrubs and other plant, also can make stub or potted landscape, be green
Change the excellent ornamental plant that beautifies the environment with floriculture at home, application prospect is huge.
Lagerstroemia indica L. can routinely adopt seminal propagation, but the differentiation of seedling pattern is big, unstable.U.S.'s red autumnal leaves Lagerstroemia indica L. production at present
On mainly by cutting propagation, cutting degree of lignification red autumnal leaves Lagerstroemia indica L. cuttage root-taking is affected very big, higher using degree of lignification
Cutting, its cuttage survival rate is high.As red autumnal leaves Lagerstroemia indica L. in the U.S. is deciduous species, after shoot sprouts, preferable cutting time is 7～
August part, cuttage production time are limited, and reproductive speed is slow, causes its nursery stock to hold at high price, and constraining its Rapid Popularization should
With.U.S.'s red autumnal leaves Lagerstroemia indica L. group culturation rapid propagating technology system that this research is set up provides effective way for its scale whole year production,
There is important theoretical and using value.Have not yet to see the tissue-culturing rapid propagation report of U.S. red autumnal leaves Lagerstroemia indica L..
By literature search, any report with regard to U.S.'s red autumnal leaves Lagerstroemia indica L. tissue-culturing rapid propagation is had not yet to see.Rarely seen a small amount of to purple
The correlational study of common vetch (Lagerstroemia indica) isolated culture.If with reference to existing Lagerstroemia indica L. tissue culture culture medium to American Red
Leaf Lagerstroemia indica L. carries out tissue-culturing rapid propagation, it may appear that leaf is fallen in the jaundice of tissue culture bud, and bud is little and weak, and elongation growth is slow, withered and yellow phenomenon finally occurs,
Situations such as effective bud number is few, causes growth coefficient little, and seedling is of poor quality, and reproduction speed is slow, and production cost is high.
Content of the invention
It is an object of the invention to overcoming shortcoming present in prior art with deficiency, there is provided a kind of U.S.'s red autumnal leaves Lagerstroemia indica L.
Tissue culture and rapid propagation method.The method has fast Induce aerosor, propagation multiplying power height, growth is fast, rooting rate is high, Seedling well developed root system of taking root
The features such as healthy and strong, transplanting survival rate height, low production cost.
The purpose of the present invention is achieved through the following technical solutions：A kind of tissue culture and rapid propagation method of U.S.'s red autumnal leaves Lagerstroemia indica L., including
Following steps：
(1) explant collection：It is female parent, clip tree to choose plant type grace, fast growth, the fine individual plant of no disease and pests harm
Dry semi-lignified branch, is cleaned up with pure water after defoliation, standby；
(2) explant sterilization and axillary bud deriving：By branch sterile water wash one time, then use alcoholic solution, mercuric chloride successively
Solution soaking is sterilized, and with inducing culture is inoculated into after aseptic water washing, is put into culturing room's culture, obtains axillary bud；
(3) enrichment culture：The axillary bud of step (2) is cut and is transferred on proliferated culture medium and is cultivated, bred
Seedling；
(4) root culture：Bud is cut in the propagation Seedling obtained from step (3) it is forwarded on root media and is cultivated,
Obtain tissue culture to take root Seedling；
(5) seedling exercising and transplanting：Step (4) tissue culture Seedling of taking root is moved on to greenhouse and carries out acclimatization and transplantses, container seedling is obtained.
In step (1), described clip butt is sprouted branch and adopts semi-lignified branch；Before collection explant, every 10d
Replace sprinkling with carbendazim, Bravo and sprout branch 4～5 times；Branch disleaf after collection, is cleaned up with pure water, low-temperature moisture preservation
Standby.
In step (2)：
Described inducing culture is ZW culture medium+0.5～1.5mg/L 6-BA+0.05～0.2mg/L IBA+30g/L
Sucrose+8g/L carrageenans；PH is 5.8-6.0；
Described ZW culture medium contains following composition：1200mg/L NH4NO3+400mg/L KNO3+450mg/L K2SO4+
440mg/L Ca(NO3)2·2H2O+76mg/L CaCl2·4H2O+260mg/L MgSO4·7H2O+170mg/L KH2PO4+
8.6mg/L ZnSO4·7H2O+6.2mg/L H3BO3+0.83mg/L KI+22.3mg/L MnSO4·4H2O+0.025mg/L
CoCl2+27.8mg/L FeSO4·7H2O+37.3mg/L Na2·EDTA·H2O+100mg/L inositol+4.0mg/L glycine+
1.2mg/L thiamine hydrochloride+2.5mg/L nicotinic acid+1.2mg/L pyridoxine hydrochloride+2.4mg/L D-VB5 calcium+2.0mg/L are anti-bad
Hematic acid, 5.8,121 DEG C of pH sterilize 20 minutes.
Described with alcoholic solution, mercuric chloride solution soaking disinfection is adopted carries out with the following method：With 70～75% ethanol of concentration
The solution soaking time is 20～30s, outwells aseptic water washing 1～2 time after alcoholic solution, is subsequently adding 0.1% mercuric chloride solution, soaks
3～10min of bubble, is constantly shaken therebetween, is outwelled after mercuric chloride solution with aseptic water washing 5～6 times.
Described culture is adopted to be carried out with the following method：In 25 ± 2 DEG C, 60 μm of ol.m-2.s-1Illumination 12h/d；Inducing culture
20d, the axillary bud of length >=1.5cm is cut and is transferred to proliferated culture medium culture；Continuous subculture 2～3 times, axillary bud is differentiated to form newly
Bud clump, the stem section that bud height >=2.0cm edible tender branch cuts into 1.0cm is transferred into proliferated culture medium culture；
Axillary bud of the described axillary bud for a height of 1～2cm of bud.
In step (3),
Described proliferated culture medium is ZW culture medium+0.5～1.5mg/L 6-BA+0.05～0.2mg/L IBA+30g/L
Sucrose+8g/L carrageenans；PH is 5.8-6.0；
Described culture is adopted to be carried out with the following method：In 25 ± 2 DEG C, 60 μm of ol.m-2.s-1Illumination 12h/d；Continuous subculture 2
～3 times, axillary bud is differentiated to form bud clump, and the tender stem of bud height >=1.5cm is cut, and accesses root media.
In step (4),
Described root media is ZW culture medium+0.1～0.5mg/L NAA+15g/L sucrose+8g/L carrageenans；PH is
5.8-6.0；
Described culture is adopted to be carried out with the following method：In 25 ± 2 DEG C, 60 μm of ol.m-2.s-1Illumination 12h/d；
Described bud is 2cm buds.
In step (5),
Described tissue culture take root Seedling be root culture 15d bottle seedling of taking root.
Described culture is adopted to be carried out with the following method：After root culture 15d, bottle seedling of taking root is moved on to
Then tissue cultured seedling is taken out by 5～7d of environment by bottle cap from standard-sized sheet seedling exercising 1d is partly reached, and cleans the culture medium sticked on root,
It is transplanted to peat soil：Perlite=3：On 1 mixed-matrix, after transplanting, then front 15d opens thin film routinely using epiphragma moisturizing
Seedling management.
The present invention is had the following advantages relative to prior art and effect：
(1) present invention is so that fast growth, plant type be graceful, pattern is gorgeous, and the U.S. red autumnal leaves Lagerstroemia indica L. of strong stress resistance grows up
Fine individual plant is female parent, to sprout branch as explant, by the optimization of minimal medium design, hormone kind, concentration and its proportioning,
Its efficiently in vitro plant regeneration technique is set up, propagation bud stalwartness elongation is reached, is grown the high fast, rate of increase, rooting rate height, takes root
Seedling is healthy and strong, the purpose that transplanting survival rate is high.The present invention have Induce aerosor is fast, bud is sturdy, elongation growth is fast, effective bud is more,
Propagation multiplying power is high, rooting rate is high, the features such as Seedling well developed root system is healthy and strong, transplanting survival rate is high of taking root, and whole technical system is stable, raw
Low cost is produced, by scale breeding, the red autumnal leaves Lagerstroemia indica L. seedling of stalwartness, neat and consistent is produced, is to realize that red autumnal leaves Lagerstroemia indica L. is quick
The demand for promoting, meeting market provides technical support and breeding guarantee, has a extensive future.
(2) U.S.'s red autumnal leaves Lagerstroemia indica L. tissue culture and rapid propagation method of the invention, the axillary bud deriving speed of explant are fast；Inductivity height,
Up to 88%；Growth coefficient is big, up to 5～6, the elongation growth of propagation bud is fast, and culture 25d buds are up to 3～4cm；Rooting rate is reachable
100%, mean elements is 4.54 pieces/plant；Tissue culture is taken root, and Seedling is healthy and strong, transplanting survival rate up to more than 95%.
Description of the drawings
Axillary bud deriving result figures of the Fig. 1 for the sterilizable material of embodiment 1；
Propagation clump bud figures of the Fig. 2 for embodiment 1；
Propagation bottle seedling figures of the Fig. 3 for embodiment 1；
Fig. 4 is the Seedling figure of taking root in the root media of embodiment 1；
Take root Seedling root system figures of the Fig. 5 for embodiment 1；
Fig. 6 is the transplanted seedling growing state figure of embodiment 1.
Specific embodiment
With reference to embodiment, the present invention is described in further detail, but embodiments of the present invention not limited to this.
Embodiment 1
A kind of tissue culture and rapid propagation method of U.S.'s red autumnal leaves Lagerstroemia indica L., comprises the steps：
(1) explant collection：It is female parent, select tree tree to choose plant type grace, fast growth, the fine individual plant of no disease and pests harm
More than 15 years age, when the fine morning 10 or so, clip trunk semi-lignified branch is cleaned with pure water after defoliation dry
Only, standby；
(2) explant sterilization and axillary bud deriving：On superclean bench, by branch sterile water wash one time, then with dense
70～75% alcoholic solution soak times of degree are 20～30s, outwell aseptic water washing 1～2 time after alcoholic solution, are subsequently adding
0.1% mercuric chloride solution, soaks 3～10min, constantly shakes therebetween, outwells with aseptic water washing 5～6 times after mercuric chloride solution, with nothing
After bacterium water is rinsed, aseptic filter paper blots sterilizable material surface water drops, stem section is cut into 2cm band axils and is cut and is inoculated into inducing culture
Base (inducing culture：ZW culture medium+0.5～1.5mg/L 6-BA+0.05～0.2mg/L IBA+30g/L sucrose+8g/L OK a karaoke clubs
Glue；PH is 5.8-6.0 (hormone is purchased from Sigma-Aldrich companies)), culturing room is put into, in 25 ± 2 DEG C, 60 μm of ol.m-2.s-1
Illumination 12h/d；Inducing culture 15d, axillary bud of the length more than 2cm is cut and is transferred to proliferated culture medium culture；Culture 30d, axil
Bud is differentiated to form sprouting clump, the stem section that bud height >=3cm edible tender branch cuts into 1.0～1.5cm is transferred and is trained into proliferated culture medium
Support；One explant of per bottle of inoculation；After 5d, petiole starts loose or dislocation, and axillary bud is sprouted, and after 15d, axillary bud can be grown left to 2cm
The right side, blade are carried out, leaf color jade green (Fig. 1), and inductivity is 88%.
(3) enrichment culture：The axillary bud for inducing of step (2) is cut into the stem section of 1.0～1.5cm and is transferred to propagation training
Foster base (ZW culture medium+0.5～1.5mg/L 6-BA+0.05～0.2mg/L IBA+30g/L sucrose+8g/L carrageenans；PH is
Cultivated on 5.8-6.0), temperature control at 25 ± 2 DEG C, illumination (60 μm of ol.m-2.s-1) time 12h/d；Subculture 3～4 times
Afterwards, simple bud forms bud clump, and bud elongation growth is fast, and growth coefficient and effective bud (bud height >=2cm) up to 5.21 and 6.2 respectively/
Clump (Fig. 2, Fig. 3).
(4) root culture：2.0cm buds are cut in the propagation Seedling obtained from step (3) is forwarded to root media (ZW cultures
Base+0.1～0.5mg/L NAA+15g/L sucrose+8g/L carrageenans；PH is 5.8-6.0) on cultivated, temperature control is 25
± 2 DEG C, illumination (60 μm of ol.m-2.s-1) time 12h/d, after 15d, the inductivity of adventitious root is 100% (Fig. 4).Robust plant,
Leaf color jade green, root system are sturdy, 4.54/plant of mean elements (Fig. 5).
(5) seedling exercising and transplanting：Step (4) tissue culture Seedling of taking root is moved on to greenhouse and carries out acclimatization and transplantses, container seedling is obtained.Will be raw
The bottle seedling of taking root of root culture 15d moves on in greenhouse adaptation 5～7d of external environment, then by bottle cap from partly reaching standard-sized sheet seedling exercising
1d, tissue cultured seedling is carefully taken out, and is cleaned the culture medium sticked on root, is transplanted to peat soil：Perlite=3：On 1 mixed-matrix,
Before after transplanting, then 15d opens thin film routinely seedling management using epiphragma moisturizing, and the transplanting survival rate is higher than 95% (Fig. 6).
In step (1), described clip butt is sprouted branch and adopts semi-lignified branch；Before collection explant, every 10d
Replace sprinkling with carbendazim, Bravo and sprout branch 4～5 times；Branch disleaf after collection, is cleaned up with pure water, low-temperature moisture preservation
Standby.
Described ZW culture medium contains following composition：1200mg/L NH4NO3+400mg/L KNO3+450mg/L K2SO4+
440mg/L Ca(NO3)2·2H2O+76mg/L CaCl2·4H2O+260mg/L MgSO4·7H2O+170mg/L KH2PO4+
8.6mg/L ZnSO4·7H2O+6.2mg/L H3BO3+0.83mg/L KI+22.3mg/L MnSO4·4H2O+0.025mg/L
CoCl2+27.8mg/L FeSO4·7H2O+37.3mg/L Na2·EDTA·H2O+100mg/L inositol+4.0mg/L glycine+
1.2mg/L thiamine hydrochloride+2.5mg/L nicotinic acid+1.2mg/L pyridoxine hydrochloride+2.4mg/L D-VB5 calcium+2.0mg/L are anti-bad
Hematic acid, 5.8,121 DEG C of pH sterilize 20 minutes.
The present invention so that fast growth, plant type be graceful, pattern is gorgeous, the U.S. red autumnal leaves Lagerstroemia indica L. of strong stress resistance adult excellent
Individual plant is female parent, to sprout branch as explant, by the optimization of minimal medium design, hormone kind, concentration and its proportioning, sets up
Its efficient in vitro plant regeneration technique, reaches the healthy and strong elongation of propagation bud, growth is fast, the rate of increase is high, rooting rate is high, Seedling of taking root is good for
The high purpose of strong, transplanting survival rate.The present invention breeds with Induce aerosor is fast, bud is sturdy, elongation growth is fast, effective bud
Multiplying power is high, rooting rate is high, the features such as Seedling well developed root system is healthy and strong, transplanting survival rate is high of taking root, and whole technical system is stable, produces into
This is low, by scale breeding, produces the red autumnal leaves Lagerstroemia indica L. seedling of stalwartness, neat and consistent, be realize red autumnal leaves Lagerstroemia indica L. Rapid Popularization,
The demand for meeting market provides technical support and breeding guarantee, has a extensive future.
In this method, the axillary bud deriving speed of explant is fast；Inductivity is high, up to 88%；Growth coefficient is big, up to 5～6, increase
Grow bud elongation growth fast, culture 25d buds are up to 3～4cm；Up to 100%, mean elements is 4.54 pieces/plant to rooting rate；Tissue culture is given birth to
Root is healthy and strong, transplanting survival rate up to more than 95%.
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention are not by above-described embodiment
Limit, other any spirit without departing from the present invention and the change, modification, replacement that is made under principle, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (8)
1. a kind of tissue culture and rapid propagation method of U.S.'s red autumnal leaves Lagerstroemia indica L., it is characterised in that comprise the steps：
(1) explant collection：It is mother to choose plant type grace, fast growth, excellent U.S.'s red autumnal leaves Lagerstroemia indica L. individual plant of no disease and pests harm
This, clip trunk semi-lignified branch is cleaned up with pure water after defoliation, standby；
(2) explant sterilization and axillary bud deriving：By branch sterile water wash one time, then use alcoholic solution, mercuric chloride solution successively
Soaking disinfection, with inducing culture is inoculated into after aseptic water washing, is put into culturing room's culture, obtains axillary bud；
(3) enrichment culture：The axillary bud of step (2) is cut and is transferred on proliferated culture medium and is cultivated, obtain breeding Seedling；
(4) root culture：Bud is cut in the propagation Seedling obtained from step (3) it is forwarded on root media and cultivated, obtains
Tissue culture is taken root Seedling；
(5) seedling exercising and transplanting：Step (4) tissue culture Seedling of taking root is moved on to greenhouse and carries out acclimatization and transplantses, container seedling is obtained；
Inducing culture described in step (2) is ZW culture medium+0.5～1.5mg/L 6-BA+0.05～0.2mg/L IBA+
30g/L sucrose+8g/L carrageenans；PH is 5.8-6.0；
Described ZW culture medium contains following composition：1200mg/L NH4NO3+400mg/L KNO3+450mg/L K2SO4+
440mg/L Ca(NO3)2·2H2O+76mg/L CaCl2·4H2O+260mg/L MgSO4·7H2O+170mg/L KH2PO4+
8.6mg/L ZnSO4·7H2O+6.2mg/L H3BO3+0.83mg/L KI+22.3mg/L MnSO4·4H2O+0.025mg/L
CoCl2+27.8mg/L FeSO4·7H2O+37.3mg/L Na2·EDTA·H2O+100mg/L inositol+4.0mg/L glycine+
1.2mg/L thiamine hydrochloride+2.5mg/L nicotinic acid+1.2mg/L pyridoxine hydrochloride+2.4mg/L D-VB5 calcium+2.0mg/L are anti-bad
Hematic acid, 5.8,121 DEG C of pH sterilize 20 minutes.
2. the tissue culture and rapid propagation method of red autumnal leaves Lagerstroemia indica L. in the U.S.'s according to claim 1, it is characterised in that described in step (2)
With alcoholic solution, mercuric chloride solution soaking disinfection is adopted carries out with the following method：With 70～75% alcoholic solution soak time of concentration
For 20～30s, aseptic water washing 1～2 time after alcoholic solution is outwelled, 0.1% mercuric chloride solution is subsequently adding, is soaked 3～10min,
Constantly shake therebetween, outwell after mercuric chloride solution with aseptic water washing 5～6 times.
3. the tissue culture and rapid propagation method of red autumnal leaves Lagerstroemia indica L. in the U.S.'s according to claim 1, it is characterised in that described in step (2)
Culture adopt and carry out with the following method：In 25 ± 2 DEG C, 60 μm of ol.m-2.s-1Illumination 12h/d；Inducing culture 20d, by length >=
The axillary bud of 1.5cm cuts and is transferred to proliferated culture medium culture；Continuous subculture 2～3 times, axillary bud is differentiated to form sprouting clump, and bud is high
>=2.0cm edible tender branch cuts into the stem section of 1.0cm and transfers into proliferated culture medium culture.
4. the tissue culture and rapid propagation method of red autumnal leaves Lagerstroemia indica L. in the U.S.'s according to claim 1, it is characterised in that described in step (2)
Axillary bud for a height of 1～2cm of bud axillary bud.
5. the tissue culture and rapid propagation method of red autumnal leaves Lagerstroemia indica L. in the U.S.'s according to claim 1, it is characterised in that described in step (3)
Culture adopt and carry out with the following method：In 25 ± 2 DEG C, 60 μm of ol.m-2.s-1Illumination 12h/d；Continuous subculture 2～3 times, axillary bud point
Change and form bud clump, the tender stem of bud height >=1.5cm is cut, access root media.
6. the tissue culture and rapid propagation method of red autumnal leaves Lagerstroemia indica L. in the U.S.'s according to claim 1, it is characterised in that described in step (4)
Root media be ZW culture medium+0.1～0.5mg/L NAA+15g/L sucrose+8g/L carrageenans；PH is 5.8-6.0.
7. the tissue culture and rapid propagation method of red autumnal leaves Lagerstroemia indica L. in the U.S.'s according to claim 1, it is characterised in that described in step (4)
Culture adopt and carry out with the following method：In 25 ± 2 DEG C, 60 μm of ol.m-2.s-1Illumination 12h/d.
8. the tissue culture and rapid propagation method of red autumnal leaves Lagerstroemia indica L. in the U.S.'s according to claim 1, it is characterised in that described in step (5)
Culture adopt and carry out with the following method：After root culture 15d, bottle seedling of taking root moves on to adaptation 5～7d of external environment in greenhouse, so
Afterwards tissue cultured seedling is taken out from standard-sized sheet seedling exercising 1d is partly reached by bottle cap, the culture medium sticked on root is cleaned, is transplanted to peat
Soil：Perlite=3：On 1 mixed-matrix, after transplanting, then front 15d opens thin film routinely seedling management using epiphragma moisturizing.
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| CN109287483B (en) * | 2018-11-02 | 2022-03-25 | 杭州市园林绿化股份有限公司 | Method for quickly cultivating hydrangea macrophylla container seedlings |
| CN109287484A (en) * | 2018-11-02 | 2019-02-01 | 杭州市园林绿化股份有限公司 | Dwarf form bigleaf hydrangea method for tissue culture |
| CN113892430B (en) * | 2021-11-15 | 2023-06-16 | 湖北格林凯尔农业科技有限公司 | Tissue culture and rapid propagation method of pinus koraiensis |
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