CN106688894A - Method for tissue culture rapid propagation of lemongrass - Google Patents
Method for tissue culture rapid propagation of lemongrass Download PDFInfo
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- CN106688894A CN106688894A CN201710046504.0A CN201710046504A CN106688894A CN 106688894 A CN106688894 A CN 106688894A CN 201710046504 A CN201710046504 A CN 201710046504A CN 106688894 A CN106688894 A CN 106688894A
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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
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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
Abstract
The invention discloses a method for tissue culture rapid propagation of lemongrass; the method comprises the processes of explant selection and treatment, initial induction, subculture, rooting culture medium and seedling hardening and transplanting and particularly comprises the steps: pruning robust lemongrass as an explant, carrying out sterilization and disinfection treatment of the explant, inoculating an initial induction culture medium with the explant to obtain an initial bud, then inoculating a subculture culture medium with the initial bud, subculturing for 15-20 days to form cluster buds, cutting off upper leaves of a single bud with the height of more than or equal to 3 cm, then transferring into a rooting culture medium, carrying out rooting induction, and finally carrying out seedling hardening and transplanting to obtain a lemongrass survival seedling. The lemongrass explant is screened by disinfection, initial induction, subculture, rooting culture and other factors and cultivation methods, appropriate seedling hardening and seedling nursery measures are adopted, a lemongrass tissue culture rapid propagation technology system is established, seed seedling traits are stable, a technical support is provided for lemongrass tissue culture factorization seedling culture, and the method has great significance in promotion of the development of lemongrass industries.
Description
Technical field
The invention belongs to plant tissue culture technical field, and in particular to a kind of lemon-grass tissue culture and rapid propagation method.
Background technology
Lemon-grass (CymbopogonCitratus grass family) is belonged to, herbaceos perennial originates in subtropical zone.
Contain citral in its cauline leaf, with citrus scented, can extract citronella oil.Citronella oil have sterilization, sterilization, relieving asthma, cough-relieving, treatment god
The medical functions such as dysmenorrhoea, courbature, are widely used in detergents and cosmetic and medical industry.Additionally, lemon-grass is developed in recent years
Tea-drinking and flavoring, can improve digestive function, and stomach invigorating disappears fat, meanwhile, lemon-grass has ornamental value, can be used to afforest, sweetening treatment
Household, and its cultivation is easy, and extensive management, water retention property is good, is a kind of water and soil conservation good grass seed with development prospect,
It is worth conducting vigorous propaganda popularization.
Lemon-grass typically uses division propagation, but this mode reproductive efficiency is very low, it is impossible to meet the need of production and market
Will, and, long-term vegetative propagation is easily accumulated and transmitted virus.Using tissue cultures and its supporting seedling-raising technique, by outer
The sport technique segments such as implant sterilization, primary induction, squamous subculture, culture of rootage and acclimatization and transplantses, with lemon-grass breeding or clone
It is material, carrying out tissue-cultured seedling factorial praluction can stablize and improve nursery stock production efficiency and quality, the scale to lemon-grass
Production is significant.
The content of the invention
Instant invention overcomes the deficiencies in the prior art, there is provided a kind of growing-seedling period is short, survival rate is high, seedling quality is good
The method of lemon-grass tissue-culturing rapid propagation.
To achieve these goals, technical scheme is as follows：
A kind of method of lemon-grass tissue-culturing rapid propagation, including explant selection and treatment, primary induction, squamous subculture, culture of rootage
Base and acclimatization and transplantses operation；Healthy and strong lemon-grass is pruned as explant, is inoculated in after carrying out sterilization treatment to explant
Obtain initial bud in primary inducing culture, then initial bud be inoculated in subculture medium by 15~20 days squamous subcultures,
Multiple Buds are formed, height >=3cm simple buds is cut and be transferred to after upper blade root induction in root media, finally carry out hardening shifting
Plant acquisition lemon-grass and survive seedling；Main operational steps are as follows：
（1）Explant is selected and processed：In the lemon-grass that fine day selection is healthy and strong, bottom residual root system and upper blade are cut off, protected
Stay stem and part leaf sheath, peel off outside 1~3 layer of leaf sheath after as explant material；Explant surface soil is rinsed out with clear water
After be put into dephosphorization detergent solution and soak 2min, while scrubbing explant with banister brush, then rinsed well with running water；
On superclean bench, explant is put into after soaking 10 s in the alcoholic solution of volumetric concentration 75%, with aseptic water washing 3 ~ 5 times,
The HgCl of volumetric concentration 0.1% is put it into again28min~12 min is soaked in solution, with aseptic water washing 3 times~5 times；
（2）Primary is induced：With scalpel by step（1）The explant for obtaining is inserted perpendicularly into primary induction after peeling 1 ~ 2 layer of leaf sheath off
In culture medium, stand steady；
（3）Squamous subculture：When stem section in primary inducing culture to be accessed grows to 5 more than cm, upper blade is cut off,
Keep initial bud sprout highly for 2.5~3.5cm, by pruning after initial bud be transferred in subculture medium, by 15~20 days
Culture, propagation forms clump bud, growth coefficient 3.0~5.0；It is simple bud or little Cong by the plant division of propagation bud, to simple bud on super-clean bench
Or little Cong proceeds squamous subculture；
（4）Culture of rootage：By step（3）The height for obtaining >=3cm simple buds cut upper blade, keep simple bud sprout height 1.5~
2.5cm, the simple bud after pruning is transferred in root media, after 7~10 days, you can root long；
（5）Acclimatization and transplantses：Treat step（4）Simple bud root length after culture of rootage moves on to outdoor nursery when reaching more than 1.0cm
Solarium carries out hardening, and culture bottle cap is opened after 3 ~ 7d, and root media and tissue-cultured seedling are poured out, and gently extrudes root media,
The rooted seedling that taking-up has been taken exercise, concussion removes residual liquor in being put into clear water；With waddy or other instruments equipped with nursery base
The container for plant growth centre-drilling hole of matter, rooted seedling is put into hole, clutches container for plant growth, and water of drenching carries out nursery stock management afterwards.
Above step（1）The length that described explant material retains stem is 2.5cm ~ 3cm.
The formula of above-described primary inducing culture is：WPM+BA 0.3mg/L~1.0 mg/L+ white sugar 30g/
L+ agar 4.5 g/L, pH=5.8.
Above-described squamous subculture based formulas are：WPM+BA 2.0mg/L~3.0 mg/L+NAA 0.2mg/L~
The g/L of 0.5 mg/L+ white sugar 3%+ agar 5.5, pH=5.8.
Above-described prescription of rooting medium is：1/2WPM+IBA 0.8mg/L~1.0 mg/L+IAA1.2mg/L
The g/L of~1.5 mg/L+ white sugar 1.5%+ agar 6.0, pH=5.8.
Above step（2）Described primary induction, step（3）Described squamous subculture, step（4）Described culture of rootage
Cultivation control condition be：Temperature is 25 ± 3 DEG C, intensity of illumination 2000Lx~8000Lx, light application time 12h/d~16h/d.
Above step（5）Described seedling medium is coconut palm chaff；Described container for plant growth specification is diameter 5cm, 10cm's high
Non-woven fabrics cup；Before transplanting, seedling medium is drenched and is disappeared with the potassium permanganate of volumetric concentration 0.5% or 800 times of thiophanate methyl solution
Poison, liquid is washed away after 1d with clear water sprinkle again.
Above step（5）Described nursery stock management method is：1 time~2 times water are sprayed daily；Transplant in one week after, printing opacity
Degree is maintained at 30%, and illumination is gradually increased after one week, transplants a month light transmittance and increases to 60~80%；Treat tissue-cultured seedling restoration ecosystem
Afterwards, with the mixing of the composite fertilizer's aqueous solution of mass concentration 0.2% or the urea of mass concentration 0.1% and the phosphate fertilizer of mass concentration 0.2%
The aqueous solution sprays blade face, and blade face is rinsed with clear water immediately after fertilising, monthly applies fertilizer 1 time, and fertilising is preferably carried out in the dusk, should not be
Applied fertilizer during noon high temperature；The N of wherein described composite fertilizer:P:K=15:15:15.
Relative to prior art, the present invention has advantages below and good effect：
1st, the present invention sterilizes by lemon-grass explant, primary induction, squamous subculture, every factor and the culture such as culture of rootage
The screening of mode, and using suitable hardening, nursery measure, lemon-grass group culturation rapid propagating technology system is established, seed and seedling traits are steady
It is fixed, for lemon-grass tissue culture industrial seedling rearing provides technical support, for promoting the development of lemongrass grassland industry significant.
2nd, the present invention is induced using healthy and strong lemon-grass as explant by primary, and initial bud is sprouted rate 85-90%；By
15-20 days squamous subcultures, form Multiple Buds, and subculture bud growth coefficient is high, up to 3.0~5.0；The simple bud of height >=3cm is passed through
Root induction in root media is inserted after pruning, acclimatization and transplantses acquisition lemon-grass is finally carried out and is survived seedling, rooting rate and transplanting
Survival rate is up to 99%, so as to shorten the growing-seedling period of lemon-grass, can in a short time obtain a large amount of tissue cultural seedlings of free, improves
The production efficiency and quality of lemon-grass.
3rd, the present invention adds corresponding according to the growing state of the different phase of lemon-grass tissue culture on WPM minimal mediums
Hormone and liquid so that culture medium is more scientific, more targetedly, are more easy to promote lemon-grass tissue-culturing rapid propagation, effect is significant.
4th, the growth characteristics cultivated under specific light and temperature condition, adapt to lemon-grass tissue-cultured seedling of the invention, promote nursery stock
Growth.
5th, nursery in matrix container of the lemon-grass tissue culture transplantation of seedlings equipped with coconut palm chaff that the present invention will be obtained, nursery material is simple
Being easy to get, and be enough to provide the nutrition of abundance promotes tissue-cultured seedling to grow, and improves transplanting survival rate.
Brief description of the drawings
Fig. 1 is lemon-grass tissue culture subculture bottle seedling of the present invention.
Fig. 2 is taken root bottle seedling for lemon-grass tissue culture of the present invention.
Fig. 3 survives seedling for lemon-grass of the present invention.
Specific embodiment
With reference to specific embodiment, the present invention will be further described.
Embodiment 1：
A kind of method of lemon-grass tissue-culturing rapid propagation, main operational steps are as follows：
（1）Explant is selected and processed：In the lemon-grass that fine day selection is healthy and strong, bottom residual root system and upper blade are cut off, protected
Stay stem and part leaf sheath, the length for retaining stem is 2.5cm ~ 3cm, peel off after 1~3 layer of leaf sheath in outside as explant material；With
Clear water to be rinsed out be put into after the soil of explant surface and soak 2min in dephosphorization detergent solution, while scrubbing explant with banister brush
Body, is then rinsed well with running water；On superclean bench, explant is put into the alcoholic solution of volumetric concentration 75% and is soaked
After steeping 10 s, with aseptic water washing 3 ~ 5 times, then the HgCl of volumetric concentration 0.1% is put it into28min is soaked in solution, with aseptic
Water is rinsed 3 times；
（2）Primary is induced：With scalpel by step（1）The explant for obtaining is inserted perpendicularly into primary induction after peeling 1 ~ 2 layer of leaf sheath off
In culture medium, stand steady；The formula of described primary inducing culture is：WPM+BA 0.3mg/L+ white sugar 30g/L+ agar
4.5 g/L, pH=5.8；The nurturing an environment of primary induction：Temperature is 25 ± 3 DEG C, intensity of illumination 2000Lx, light application time 16h/d；
Initial bud rate of sprouting is 80%；
（3）Squamous subculture：When stem section in primary inducing culture to be accessed grows to 5 more than cm, upper blade is cut off,
Keep initial bud sprout highly for 2.5~3.5cm, by pruning after initial bud be transferred in subculture medium, by 15~20 days
Culture, propagation forms clump bud, growth coefficient 3.0~5.0；It is simple bud or little Cong by the plant division of propagation bud, to simple bud on super-clean bench
Or little Cong proceeds squamous subculture；Described squamous subculture based formulas are：WPM+BA 2.0mg/L+NAA 0.2mg/L~
The g/L of 0.5 mg/L+ white sugar 3%+ agar 5.5, pH=5.8, wherein the amount of the white sugar for being added is culture medium quality
3%；The nurturing an environment of squamous subculture：Temperature is 25 ± 3 DEG C, intensity of illumination 2000Lx, light application time 16h/d；
（4）Culture of rootage：By step（3）The height for obtaining >=3cm simple buds cut upper blade, keep simple bud sprout height 1.5~
2.0cm, the simple bud after pruning is transferred in root media, after 7~10 days, you can root long；Described prescription of rooting medium
For：The g/L of 1/2WPM+IBA 0.8mg/L+IAA1.2mg/L+ white sugar 1.5%+ agar 6.0, pH=5.8, wherein added
The amount of the white sugar for entering is the 1.5% of culture medium quality；The nurturing an environment of culture of rootage：Temperature is 25 ± 3 DEG C, intensity of illumination
2000Lx, light application time 16h/d；Rooting rate is 99%；
（5）Acclimatization and transplantses：Treat step（4）Simple bud root length after culture of rootage moves on to outdoor nursery when reaching more than 1.0cm
Solarium carries out hardening, and culture bottle cap is opened after 3 ~ 7d, and root media and tissue-cultured seedling are poured out, and gently extrudes root media,
The rooted seedling that taking-up has been taken exercise, concussion removes residual liquor in being put into clear water；With waddy or other instruments equipped with coconut palm chaff
Diameter 5cm, 10cm high non-woven fabrics cup centre-drilling hole, rooted seedling is put into hole, clutch non-woven fabrics cup, water of drenching, afterwards
Carry out nursery stock management.Before transplanting, coconut palm chaff is drenched and is disappeared with the potassium permanganate of volumetric concentration 0.5% or 800 times of thiophanate methyl solution
Poison, liquid is washed away after 1d with clear water sprinkle again.Transplanting survival rate is 99%.
Described nursery stock management method is：1 time~2 times water are sprayed daily；Transplant in one week after, light transmittance is maintained at 30%,
Gradually increase illumination after one week, transplant a month light transmittance and increase to 60%；After after tissue-cultured seedling restoration ecosystem, with mass concentration 0.2%
Composite fertilizer's aqueous solution sprays blade face, and blade face is rinsed with clear water immediately after fertilising, monthly applies fertilizer 1 time, and fertilising is preferably carried out in the dusk,
Should not be applied fertilizer in noon high temperature；The N of wherein described composite fertilizer:P:K=15:15:15.
Embodiment 2：
A kind of method of lemon-grass tissue-culturing rapid propagation, main operational steps are as follows：
（1）Explant is selected and processed：In the lemon-grass that fine day selection is healthy and strong, bottom residual root system and upper blade are cut off, protected
Stay stem and part leaf sheath, the length for retaining stem is 2.5cm ~ 3cm, peel off after 1~3 layer of leaf sheath in outside as explant material；With
Clear water to be rinsed out be put into after the soil of explant surface and soak 2min in dephosphorization detergent solution, while scrubbing explant with banister brush
Body, is then rinsed well with running water；On superclean bench, explant is put into the alcoholic solution of volumetric concentration 75% and is soaked
After steeping 10 s, with aseptic water washing 3 ~ 5 times, then the HgCl of volumetric concentration 0.1% is put it into210 min are soaked in solution, with nothing
Bacterium water is rinsed 4 times；
（2）Primary is induced：With scalpel by step（1）The explant for obtaining is inserted perpendicularly into primary induction after peeling 1 ~ 2 layer of leaf sheath off
In culture medium, stand steady；The formula of described primary inducing culture is：The mg/L+ white sugar 30g/L+ agar of WPM+BA 0.5
4.5 g/L, pH=5.8；The nurturing an environment of primary induction：Temperature is 25 ± 3 DEG C, intensity of illumination 4000Lx, light application time 14h/d；
Initial bud rate of sprouting is 85%；
（3）Squamous subculture：When stem section in primary inducing culture to be accessed grows to 5 more than cm, upper blade is cut off,
Keep initial bud sprout highly for 2.5~3.0cm, by pruning after initial bud be transferred in subculture medium, by 15~20 days
Culture, propagation forms clump bud, growth coefficient 3.0~5.0；It is simple bud or little Cong by the plant division of propagation bud, to simple bud on super-clean bench
Or little Cong proceeds squamous subculture；Described squamous subculture based formulas are：The mg/L+NAA 0.2mg/L of WPM+BA 2.5~
The g/L of 0.5 mg/L+ white sugar 3%+ agar 5.5, pH=5.8, wherein the amount of the white sugar for being added is culture medium quality
3%；The nurturing an environment of squamous subculture：Temperature is 25 ± 3 DEG C, intensity of illumination 4000Lx, light application time 14h/d；
（4）Culture of rootage：By step（3）The height for obtaining >=3cm simple buds cut upper blade, keep simple bud sprout height 1.5~
2.0cm, the simple bud after pruning is transferred in root media, after 7~10 days, you can root long；Described prescription of rooting medium
For：The g/L of 1/2WPM+IBA 0.8mg/L+IAA1.4 mg/L+ white sugar 1.5%+ agar 6.0, pH=5.8, wherein added
The amount of the white sugar for entering is the 1.5% of culture medium quality；The nurturing an environment of culture of rootage：Temperature is 25 ± 3 DEG C, intensity of illumination
4000Lx, light application time 14h/d；Rooting rate is 99%；
（5）Acclimatization and transplantses：Treat step（4）Simple bud root length after culture of rootage moves on to outdoor nursery when reaching more than 1.0cm
Solarium carries out hardening, and culture bottle cap is opened after 3 ~ 7d, and root media and tissue-cultured seedling are poured out, and gently extrudes root media,
The rooted seedling that taking-up has been taken exercise, concussion removes residual liquor in being put into clear water；With waddy or other instruments equipped with coconut palm chaff
Diameter 5cm, 10cm high non-woven fabrics cup centre-drilling hole, rooted seedling is put into hole, clutch non-woven fabrics cup, water of drenching, afterwards
Carry out nursery stock management.Before transplanting, coconut palm chaff is drenched and is disappeared with the potassium permanganate of volumetric concentration 0.5% or 800 times of thiophanate methyl solution
Poison, liquid is washed away after 1d with clear water sprinkle again.Transplanting survival rate is 99%.
Described nursery stock management method is：1 time~2 times water are sprayed daily；Transplant in one week after, light transmittance is maintained at 30%,
Gradually increase illumination after one week, transplant a month light transmittance and increase to 70%；After after tissue-cultured seedling restoration ecosystem, user's mass concentration
The mixed aqueous solution of 0.1% urea and the phosphate fertilizer of mass concentration 0.2% sprays blade face, rinses blade face with clear water immediately after fertilising, often
The moon is applied fertilizer 1 time, and fertilising is preferably carried out in the dusk, should not be applied fertilizer in noon high temperature；The N of wherein described composite fertilizer:P:K=15:
15:15。
Embodiment 3：
A kind of method of lemon-grass tissue-culturing rapid propagation, main operational steps are as follows：
（1）Explant is selected and processed：In the lemon-grass that fine day selection is healthy and strong, bottom residual root system and upper blade are cut off, protected
Stay stem and part leaf sheath, the length for retaining stem is 2.5cm ~ 3cm, peel off after 1~3 layer of leaf sheath in outside as explant material；With
Clear water to be rinsed out be put into after the soil of explant surface and soak 2min in dephosphorization detergent solution, while scrubbing explant with banister brush
Body, is then rinsed well with running water；On superclean bench, explant is put into the alcoholic solution of volumetric concentration 75% and is soaked
After steeping 10 s, with aseptic water washing 3 ~ 5 times, then the HgCl of volumetric concentration 0.1% is put it into212 min are soaked in solution, with nothing
Bacterium water is rinsed 5 times；
（2）Primary is induced：With scalpel by step（1）The explant for obtaining is inserted perpendicularly into primary induction after peeling 1 ~ 2 layer of leaf sheath off
In culture medium, stand steady；The formula of described primary inducing culture is：The mg/L+ white sugar 30g/L+ agar of WPM+BA 1.0
4.5 g/L, pH=5.8；The nurturing an environment of primary induction：Temperature is 25 ± 3 DEG C, intensity of illumination 8000Lx, light application time 12h/d；
Initial bud rate of sprouting is 90%；
（3）Squamous subculture：When stem section in primary inducing culture to be accessed grows to 5 more than cm, upper blade is cut off,
Keep initial bud sprout highly for 3.0~3.5cm, by pruning after initial bud be transferred in subculture medium, by 15~20 days
Culture, propagation forms clump bud, growth coefficient 3.0~5.0；It is simple bud or little Cong by the plant division of propagation bud, to simple bud on super-clean bench
Or little Cong proceeds squamous subculture；Described squamous subculture based formulas are：The mg/L+NAA 0.2mg/L of WPM+BA 3.0~
The g/L of 0.5 mg/L+ white sugar 3%+ agar 5.5, pH=5.8, wherein the amount of the white sugar for being added is culture medium quality
3%；The nurturing an environment of squamous subculture：Temperature is 25 ± 3 DEG C, intensity of illumination 8000Lx, light application time 12h/d；
（4）Culture of rootage：By step（3）The height for obtaining >=3cm simple buds cut upper blade, keep simple bud sprout height 2.0~
2.5cm, the simple bud after pruning is transferred in root media, after 7~10 days, you can root long；Described prescription of rooting medium
For：The g/L of 1.0 mg/L+IAA1.5 mg/L+ white sugar 1.5%+ agar of 1/2WPM+IBA 6.0, wherein pH=5.8, institute
The amount of the white sugar of addition is the 1.5% of culture medium quality；The nurturing an environment of culture of rootage：Temperature is 25 ± 3 DEG C, intensity of illumination
8000Lx, light application time 12h/d；Rooting rate is 99%；
（5）Acclimatization and transplantses：Treat step（4）Simple bud root length after culture of rootage moves on to outdoor nursery when reaching more than 1.0cm
Solarium carries out hardening, and culture bottle cap is opened after 3 ~ 7d, and root media and tissue-cultured seedling are poured out, and gently extrudes root media,
The rooted seedling that taking-up has been taken exercise, concussion removes residual liquor in being put into clear water；With waddy or other instruments equipped with coconut palm chaff
Diameter 5cm, 10cm high non-woven fabrics cup centre-drilling hole, rooted seedling is put into hole, clutch non-woven fabrics cup, water of drenching, afterwards
Carry out nursery stock management.Before transplanting, coconut palm chaff is drenched and is disappeared with the potassium permanganate of volumetric concentration 0.5% or 800 times of thiophanate methyl solution
Poison, liquid is washed away after 1d with clear water sprinkle again.Transplanting survival rate is 99%.
Described nursery stock management method is：1 time~2 times water are sprayed daily；Transplant in one week after, light transmittance is maintained at 30%,
Gradually increase illumination after one week, transplant a month light transmittance and increase to 80%；After after tissue-cultured seedling restoration ecosystem, user's mass concentration
The mixed aqueous solution of 0.1% urea and the phosphate fertilizer of mass concentration 0.2% sprays blade face, rinses blade face with clear water immediately after fertilising, often
The moon is applied fertilizer 1 time, and fertilising is preferably carried out in the dusk, should not be applied fertilizer in noon high temperature；The N of wherein described composite fertilizer:P:K=15:
15:15。
Claims (8)
1. a kind of method of lemon-grass tissue-culturing rapid propagation, it is characterised in that：Selected including explant and treatment, primary induction, subculture
Culture, root media and acclimatization and transplantses operation；Healthy and strong lemon-grass is pruned as explant, sterilization is carried out to explant
It is inoculated in after treatment in primary inducing culture and obtains initial bud, then initial bud is inoculated in subculture medium by 15~20
Its squamous subculture, forms Multiple Buds, height >=3cm simple buds is cut and be transferred to after upper blade root induction in root media, most
After carry out acclimatization and transplantses obtain lemon-grass survive seedling；Main operational steps are as follows：
（1）Explant is selected and processed：In the lemon-grass that fine day selection is healthy and strong, bottom residual root system and upper blade are cut off, protected
Stay stem and part leaf sheath, peel off outside 1~3 layer of leaf sheath after as explant material；Explant surface soil is rinsed out with clear water
After be put into dephosphorization detergent solution and soak 2min, while scrubbing explant with banister brush, then rinsed well with running water；
On superclean bench, explant is put into after soaking 10 s in the alcoholic solution of volumetric concentration 75%, with aseptic water washing 3 ~ 5 times,
The HgCl of volumetric concentration 0.1% is put it into again28min~12 min is soaked in solution, with aseptic water washing 3 times~5 times；
（2）Primary is induced：With scalpel by step（1）The explant for obtaining is inserted perpendicularly into primary induction after peeling 1 ~ 2 layer of leaf sheath off
In culture medium, stand steady；
（3）Squamous subculture：When stem section in primary inducing culture to be accessed grows to 5 more than cm, upper blade is cut off,
Keep initial bud sprout highly for 2.5~3.5cm, by pruning after initial bud be transferred in subculture medium, by 15~20 days
Culture, propagation forms clump bud, growth coefficient 3.0~5.0；It is simple bud or little Cong by the plant division of propagation bud, to simple bud on super-clean bench
Or little Cong proceeds squamous subculture；
（4）Culture of rootage：By step（3）The height for obtaining >=3cm simple buds cut upper blade, keep simple bud sprout height 1.5~
2.5cm, the simple bud after pruning is transferred in root media, after 7~10 days, you can root long；
（5）Acclimatization and transplantses：Treat step（4）Simple bud root length after culture of rootage moves on to outdoor nursery when reaching more than 1.0cm
Solarium carries out hardening, and culture bottle cap is opened after 3 ~ 7d, and root media and tissue-cultured seedling are poured out, and gently extrudes root media,
The rooted seedling that taking-up has been taken exercise, concussion removes residual liquor in being put into clear water；With waddy or other instruments equipped with nursery base
The container for plant growth centre-drilling hole of matter, rooted seedling is put into hole, clutches container for plant growth, and water of drenching carries out nursery stock management afterwards.
2. the method for a kind of lemon-grass tissue-culturing rapid propagation according to claim 1, it is characterised in that：Step（1）Described is outer
The length that implant material retains stem is 2.5cm ~ 3cm.
3. the method for a kind of lemon-grass tissue-culturing rapid propagation according to claim 1, it is characterised in that：Described primary induction training
Support base formula be：WPM+BA 0.3mg/L~1.0 mg/L+ white sugar 30g/L+ agar 4.5 g/L, pH=5.8.
4. the method for a kind of lemon-grass tissue-culturing rapid propagation according to claim 1, it is characterised in that：Described subculture medium
It is formulated and is：The g/ of WPM+BA 2.0mg/L~3.0 mg/L+NAA 0.2mg/L~0.5 mg/L+ white sugar 3%+ agar 5.5
L, pH=5.8.
5. the method for a kind of lemon-grass tissue-culturing rapid propagation according to claim 1, it is characterised in that：Described root media
It is formulated and is：1/2WPM+IBA 0.8mg/L~1.0 mg/L+IAA1.2mg/L~1.5 mg/L+ white sugar 1.5%+ agar
6.0 g/L, pH=5.8.
6. the method for a kind of lemon-grass tissue-culturing rapid propagation according to claim 1, it is characterised in that：Step（2）Described is first
Generation induction, step（3）Described squamous subculture, step（4）The cultivation control condition of described culture of rootage is：Temperature is 25
± 3 DEG C, intensity of illumination 2000Lx~8000Lx, light application time 12h/d~16h/d.
7. the method for a kind of lemon-grass tissue-culturing rapid propagation according to claim 1, it is characterised in that：Step（5）Described educates
Seedling matrix is coconut palm chaff；Described container for plant growth specification is the non-woven fabrics cup of diameter 5cm, 10cm high；Before transplanting, volumetric concentration is used
0.5% potassium permanganate or 800 times of thiophanate methyl solution are drenched sterilization to seedling medium, and liquid is washed away with clear water sprinkle again after 1d.
8. the method for a kind of lemon-grass tissue-culturing rapid propagation according to claim 1, it is characterised in that：Step（5）Described seedling
Wooden management method is：1 time~2 times water are sprayed daily；Transplant in one week after, light transmittance is maintained at 30%, and light is gradually increased after one week
According to one month light transmittance of transplanting increases to 60~80%；After after tissue-cultured seedling restoration ecosystem, with the composite fertilizer's aqueous solution of mass concentration 0.2%
Or the mixed aqueous solution of the urea of mass concentration 0.1% and the phosphate fertilizer of mass concentration 0.2% sprays blade face, uses clear water after fertilising immediately
Blade face is rinsed, is monthly applied fertilizer 1 time, and fertilising is preferably carried out in the dusk, should not be applied fertilizer in noon high temperature；Wherein described composite fertilizer
N:P:K=15:15:15.
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Application publication date: 20170524 Assignee: Guangxi Tengxian Ranheli Agricultural Trade Co.,Ltd. Assignor: GUANGXI ZHUANG AUTONOMOUS REGION FORESTRY Research Institute Contract record no.: X2022450000421 Denomination of invention: A method of tissue culture and rapid propagation of citronella Granted publication date: 20190201 License type: Common License Record date: 20221227 |