CN109729976A - A kind of floral leaf fernleaf hedge bamboo tissue culture and rapid propagation method - Google Patents
A kind of floral leaf fernleaf hedge bamboo tissue culture and rapid propagation method Download PDFInfo
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- CN109729976A CN109729976A CN201910133402.1A CN201910133402A CN109729976A CN 109729976 A CN109729976 A CN 109729976A CN 201910133402 A CN201910133402 A CN 201910133402A CN 109729976 A CN109729976 A CN 109729976A
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Abstract
A kind of floral leaf fernleaf hedge bamboo tissue culture and rapid propagation method, belongs to tissue culture technology field.It is comprised the following steps that: the tender floral leaf fernleaf hedge bamboo stem-segment with node of acquisition field children is as explant；Floral leaf fernleaf hedge bamboo explant is sterilized and is cleaned；It is seeded in aseptic culture medium and cultivates；Multiplying culture；Culture of rootage；Transplanting domestication.The present invention is by realizing floral leaf fernleaf hedge bamboo and quickly breeding to floral leaf fernleaf hedge bamboo acquisition time, disinfection way, proliferated culture medium, root media and the restriction for transplanting acclimation method.The explant survival rate obtained by means of the present invention is up to 58.8%；Clump bud inductivity is up to 71.67%；Cultivation effect is up to 3.65 times；For rooting rate up to 70% or more, condition of rooting is good；Acclimation method involved in the present invention shortens more than half than traditional single plant bagging acclimation method required time, and seedling quality is unaffected.
Description
Technical field
The invention belongs to tissue culture technology fields, and in particular to a kind of floral leaf fernleaf hedge bamboo tissue culture and rapid propagation method.
Background technique
Floral leaf fernleaf hedge bamboo is the mutation of excellent ornament bamboo fernleaf hedge bamboo, belongs to grass family Bambusoideae, for bamboo kind of growing thickly.Its leaf is normal
Have irregular white vertical stripe, have more ornamental value compared with fernleaf hedge bamboo, but passes through plant division or the vegetative propagation of cuttage all the time
Mode is bred, and practical problem slow, at high cost that there are speed leads to not meet bamboo seedling market to floral leaf fernleaf hedge bamboo seedling
Demand.Using tissue rapid propagation technology, floral leaf fernleaf hedge bamboo can be carried out a large amount of and quickly bred, but so far there are no mature flower
The Explants In Tissue Culture fast breeding technique of leaf fernleaf hedge bamboo.
Summary of the invention
In view of the problems of the existing technology, it is an object of the invention to design to provide a kind of floral leaf phoenix tail bamboo tissue culture rapid breeding
The technical solution of method.
A kind of floral leaf fernleaf hedge bamboo tissue culture and rapid propagation method, it is characterised in that comprise the following steps that:
1) in the tender floral leaf fernleaf hedge bamboo stem-segment with node of late June acquisition field children as explant；
2) sodium hypochlorite of the floral leaf fernleaf hedge bamboo explant effective chlorine density 0.5~1.5% obtained to step 1) is vacuumizing item
10~14min is sterilized under part, then uses sterile water wash；
3) the stalk sheaths of bamboo shoots for aseptically peelling off the floral leaf fernleaf hedge bamboo explant that step 2 obtains, takes and ties 0.5 cm, forge 0.5
The stem with bud of cm is seeded in aseptic culture medium after blotting surface moisture with aseptic filter paper and cultivates；
4) take sterile, well-grown bud that step 3) obtains, be seeded in proliferated culture medium and cultivate, the proliferated culture medium with
MS culture medium is basic culture medium, adds 30 g/L sucrose, 3.5 g/L Gelrite, 0.08~2 mg/L TDZ, 1~4mg/L
BA and 0.2~5 mg/L KT, pH 5.7；
5) it is one clump that the clump bud for taking step 4) to obtain, which is divided into 3-5, is seeded in root media and cultivates, the root media
It is basic culture medium with MS culture medium, adds 30g/L sucrose, 3.5g/L Gelrite and 0.1~2.5mg/L NAA, pH
5.7；
6) the floral leaf fernleaf hedge bamboo plant to have taken root for taking step 5) to obtain carries out transplanting domestication.
A kind of floral leaf fernleaf hedge bamboo tissue culture and rapid propagation method, it is characterised in that floral leaf fernleaf hedge bamboo explant in the step 2
Body sterilizes 12min with the sodium hypochlorite of effective chlorine density 1% under vacuum-pumping conditions.
A kind of floral leaf fernleaf hedge bamboo tissue culture and rapid propagation method, it is characterised in that the step 3), 4) and 5) in cultivate item
Part are as follows: intensity of illumination 2500lx, daily light application time are 16 hours, and environment temperature is 25 ± 2 DEG C, relative air humidity 60
~90%.
A kind of floral leaf fernleaf hedge bamboo tissue culture and rapid propagation method, it is characterised in that proliferated culture medium is in the step 4) with MS
Culture medium is basic culture medium, 30 g/L sucrose of addition, 3.5 g/L Gelrite, 0.08mg/L TDZ, 1mg/L BA and
0.2mg/L KT, pH 5.7.
A kind of floral leaf fernleaf hedge bamboo tissue culture and rapid propagation method, it is characterised in that root media is in the step 5) with MS
Culture medium is basic culture medium, adds 30g/L sucrose, 3.5g/L Gelrite and 2.5mg/L NAA, pH 5.7.
A kind of floral leaf fernleaf hedge bamboo tissue culture and rapid propagation method, it is characterised in that transplanting domestication is specific in the step 6)
Are as follows: it is 20000 lx that the floral leaf fernleaf hedge bamboo plant to have taken root, which is moved to intensity of illumination, and daily light application time is 16 hours, environment
Temperature is 25 ± 2 DEG C, after taming 1 week under the environmental condition that relative air humidity is 70~95%, removes bottle cap and continues training 3 days,
Then the culture medium being attached on plant root is washed away with 35~40 DEG C of warm water, 3~5 points is impregnated in 1000 times of carbendazols
Clock, transplanting to the perlite stirred with 1000 times of carbendazols: peat: in vermiculite=1:1:1 matrix, and keeping wet, and 1
Domestication terminates to can be moved to greenhouse after month.
The present invention is by taming and dociling floral leaf fernleaf hedge bamboo acquisition time, disinfection way, proliferated culture medium, root media and transplanting
The restriction of change method realizes floral leaf fernleaf hedge bamboo and quickly breeds.The explant survival rate highest obtained by means of the present invention
Up to 58.8%；Clump bud inductivity is up to 71.67%；Cultivation effect is up to 3.65 times；For rooting rate up to 70% or more, condition of rooting is good；
Acclimation method involved in the present invention than shortening more than half the time required to traditional single plant bagging acclimation method, seedling quality not by
It influences.
Detailed description of the invention
Fig. 1 difference sodium hypochlorite concentration, takes explant time and stalk sheaths of bamboo shoots to retain the dirt for handling lower explant at disinfecting time
Dye rate and survival.
Specific embodiment
Further illustrate the present invention with reference to embodiments.
A kind of embodiment: floral leaf fernleaf hedge bamboo tissue culture and rapid propagation method
1) in the tender floral leaf fernleaf hedge bamboo stem-segment with node of late June acquisition field children as explant (2-5 cm, forges 1cm on section)；
2) sodium hypochlorite of the floral leaf fernleaf hedge bamboo explant effective chlorine density 0.5~1.5% obtained to step 1) is vacuumizing item
10~14min is sterilized under part, then uses sterile water wash；
3) the stalk sheaths of bamboo shoots for aseptically peelling off the floral leaf fernleaf hedge bamboo explant that step 2 obtains, takes and ties 0.5 cm, forge 0.5
The stem with bud of cm is seeded to culture in aseptic culture medium (MS culture medium), cultivates item after blotting surface moisture with aseptic filter paper
Part are as follows: intensity of illumination 2500lx, daily light application time are 16 hours, and environment temperature is 25 ± 2 DEG C, relative air humidity 60
~90%, it cultivates 30 days；
4) take sterile, well-grown bud that step 3) obtains, be seeded in proliferated culture medium and cultivate, the proliferated culture medium with
MS culture medium is basic culture medium, adds 30 g/L sucrose, 3.5 g/L Gelrite, 0.08~2 mg/L TDZ, 1~4mg/L
BA and 0.2~5 mg/L KT, pH 5.7, condition of culture are as follows: condition of culture are as follows: intensity of illumination 2500lx, when daily illumination
Between be 16 hours, environment temperature be 25 ± 2 DEG C, relative air humidity be 60~90%, cultivate 16 days；
5) it is one clump that the clump bud for taking step 4) to obtain, which is divided into 3-5, is seeded in root media and cultivates, the root media
It is basic culture medium with MS culture medium, adds 30g/L sucrose, 3.5g/L Gelrite(crystal agar) and 0.1~2.5mg/L
NAA, pH 5.7, condition of culture are as follows: condition of culture are as follows: intensity of illumination 2500lx, daily light application time are 16 hours, environment
Temperature is 25 ± 2 DEG C, and relative air humidity is 60~90%, is cultivated 16 days；
6) the floral leaf fernleaf hedge bamboo plant to have taken root for taking step 5) to obtain carries out transplanting domestication, the flower that will specially take root
It is 20000 lx that leaf fernleaf hedge bamboo plant, which moves to intensity of illumination, and daily light application time is 16 hours, and environment temperature is 25 ± 2 DEG C, empty
After taming 1 week under the environmental condition that gas relative humidity is 70~95%, removes bottle cap and continue training 3 days, then with 35~40 DEG C
Warm water washes away the culture medium being attached on plant root, impregnates 3~5 minutes in 1000 times of carbendazols, and transplanting is extremely with 1000
The perlite of times carbendazol stirring: peat: in vermiculite=1:1:1 matrix, and keep wet, domestication terminates after 1 month
Move to greenhouse.
Test example:
1) acquisition of aseptic explant
A: in 6 months, taking the tender floral leaf fernleaf hedge bamboo stem-segment with node (2-5 cm, forges 1cm on section) of field children, do not peel off stalk sheaths of bamboo shoots,
After 2 h of flowing water undershoot, first with 75% alcohol disinfecting, 30 s, effective chlorine density 0.5%, 1% and are used respectively after sterile washing 3~5 times
1.5% liquor natrii hypochloritis sterilizes 10 min, sterile washing 5~7 times under vacuum-pumping conditions.Aseptically peel off stalk
Sheaths of bamboo shoots takes and ties 0.5 cm, forges the stem with bud of 0.5 cm, is blotted with aseptic filter paper and is seeded to aseptic culture medium after showing moisture
In；Be placed on intensity of illumination be 2500lx, daily light application time be 16 hours, environment temperature be 25 ± 2 DEG C, air is relatively wet
Degree is cultivated 30 days under conditions of being 60~90%；
B: the tender floral leaf fernleaf hedge bamboo stem-segment with node of field children by the end of June, is being taken, is being taken out with the liquor natrii hypochloritis that effective chlorine density is 1%
8,10,12 and 14min is sterilized under vacuum condition respectively, and stalk sheaths of bamboo shoots is peelled off to time for being 1% with effective chlorine density before bath
Sodium chlorate solution sterilizes 10 and 12min under vacuum-pumping conditions, remaining is identical as method in A.
C: in 7 months, the tender floral leaf fernleaf hedge bamboo stem-segment with node of field children, the liquor natrii hypochloritis for being 1% with effective chlorine density are taken
10min is sterilized under vacuum-pumping conditions, remaining is identical as method in A.
It is calculated as pollution rate to pollute the ratio of number and total inoculation number, with sum uncontaminated and that budding can be sprouted and always inoculation
Several ratio is calculated as survival rate, the result is shown in Figure 1.
(2) clump bud induction and fast breeding
The aseptic explant obtained by (1) is seeded in clump bud induction and proliferated culture medium, clump bud induction and Multiplying culture
It is basic culture medium that base, which is with MS culture medium, adds 30 g/L sucrose, 3.5 g/L Gelrite, then add BA(1,2,4mg/
L), KT(0.2,1,5mg/L) and TDZ(0.08,0.4,2mg/L), it (is specifically shown in Table using 9 kinds of culture mediums of orthogonal
1).It is 2500lx that inoculation, which is placed on intensity of illumination, and daily light application time is 16 hours, and environment temperature is 25 ± 2 DEG C, and air is opposite
Humidity is cultivated under conditions of being 60~90%.It the results are shown in Table 1.
(3) root induction
Clump bud after proliferation is divided into 3~6 one clump to be seeded in root media, root media is based on MS culture medium
Culture medium adds 30g/L sucrose, 3.5g/L Gelrite, then adds NAA (0.1,0.5,2.5mg/L), and inoculation is placed on light
It is 2500lx according to intensity, daily light application time is 16 hours, and environment temperature is 25 ± 2 DEG C, and relative air humidity is 60~90%
Under the conditions of cultivate.It the results are shown in Table 2.
(4) transplanting domestication
It is 20000 lx that the floral leaf fernleaf hedge bamboo plant to have taken root, which is moved to intensity of illumination, and daily light application time is 16 hours, ring
Border temperature is 25 ± 2 DEG C, after taming 1 week under the environmental condition that relative air humidity is 70~95%, removes bottle cap and continues training 3
It, then washes away the culture medium being attached on plant root with 35~40 DEG C of warm water, in 1000 times of carbendazols impregnate 3~
5 minutes, transplanting to the perlite stirred with 1000 times of carbendazols: peat: in vermiculite=1:1:1 matrix, and keeping wet, and 1
Domestication terminates to can be moved to greenhouse after a month.
The proliferation of inductivity and fast breeding that 1 BA, KT and TDZ orthogonal test 9 of table processing floral leaf fernleaf hedge bamboo induces from bud
Coefficient
The different minimal mediums of table 2 and different NAA concentration cooperate the situation of taking root of lower floral leaf fernleaf hedge bamboo test tube seedling
。
By Fig. 1 and table 1,2 comprehensive analysis it is found that being by the end of June the Best Times for taking explant；With time of effective chlorine density 1%
Sodium chlorate sterilizes 12 min under vacuum-pumping conditions to obtain aseptic explant as best disinfection way； MS+30 g/L
+ 0.2 mg/L KT, pH 5.7 of Sucrose+3.5 g/L Gelrite+0.08 mg/L TDZ+1 mg/L BA is quickly high
Imitate the culture medium prescription of proliferation；MS++30g/L Sucrose+3.5g/L Gelrite+2.5mg/L NAA, pH 5.7 is to take root
Culture medium prescription.
Compared with other processing, of the invention most preferably takes the explant survival rate of explant body method to be up to 58.8%；Clump bud
Inductivity is up to 71.67%；Cultivation effect is up to 3.65 times；For rooting rate up to 70% or more, condition of rooting is good.
The method that the present invention passes through quick breeding by group culture floral leaf fernleaf hedge bamboo, is obtaining a certain number of floral leaf phoenix tails by the end of June
After the sterile tissue-cultured seedling of bamboo, annual quick breeding can be carried out indoors, and the test tube seedling taken root can get excellent after domestication
Floral leaf fernleaf hedge bamboo seedling, acclimation method in the present invention than the shortening of traditional single plant bagging acclimation method required time more than half,
And seedling quality is unaffected.
Claims (6)
1. a kind of floral leaf fernleaf hedge bamboo tissue culture and rapid propagation method, it is characterised in that comprise the following steps that:
1) in the tender floral leaf fernleaf hedge bamboo stem-segment with node of late June acquisition field children as explant；
2) sodium hypochlorite of the floral leaf fernleaf hedge bamboo explant effective chlorine density 0.5~1.5% obtained to step 1) is vacuumizing item
10~14min is sterilized under part, then uses sterile water wash；
3) the stalk sheaths of bamboo shoots for aseptically peelling off the floral leaf fernleaf hedge bamboo explant that step 2 obtains, takes and ties 0.5 cm, forge 0.5
The stem with bud of cm is seeded in aseptic culture medium after blotting surface moisture with aseptic filter paper and cultivates；
4) take sterile, well-grown bud that step 3) obtains, be seeded in proliferated culture medium and cultivate, the proliferated culture medium with
MS culture medium is basic culture medium, adds 30 g/L sucrose, 3.5 g/L Gelrite, 0.08~2 mg/L TDZ, 1~4mg/L
BA and 0.2~5 mg/L KT, pH 5.7；
5) it is one clump that the clump bud for taking step 4) to obtain, which is divided into 3-5, is seeded in root media and cultivates, the root media
It is basic culture medium with MS culture medium, adds 30g/L sucrose, 3.5g/L Gelrite and 0.1~2.5mg/L NAA, pH
5.7；
6) the floral leaf fernleaf hedge bamboo plant to have taken root for taking step 5) to obtain carries out transplanting domestication.
2. a kind of floral leaf fernleaf hedge bamboo tissue culture and rapid propagation method as described in claim 1, it is characterised in that floral leaf in the step 2
Fernleaf hedge bamboo explant sterilizes 12min with the sodium hypochlorite of effective chlorine density 1% under vacuum-pumping conditions.
And 5) 3. a kind of floral leaf fernleaf hedge bamboo tissue culture and rapid propagation method as described in claim 1, it is characterised in that the step 3), 4)
Middle condition of culture are as follows: intensity of illumination 2500lx, daily light application time are 16 hours, and environment temperature is 25 ± 2 DEG C, and air is opposite
Humidity is 60~90%.
4. a kind of floral leaf fernleaf hedge bamboo tissue culture and rapid propagation method as described in claim 1, it is characterised in that be proliferated in the step 4)
Culture medium is basic culture medium with MS culture medium, adds 30 g/L sucrose, 3.5 g/L Gelrite, 0.08mg/L TDZ, 1mg/
L BA and 0.2mg/L KT, pH 5.7.
5. a kind of floral leaf fernleaf hedge bamboo tissue culture and rapid propagation method as described in claim 1, it is characterised in that take root in the step 5)
Culture medium is basic culture medium with MS culture medium, adds 30g/L sucrose, 3.5g/L Gelrite and 2.5mg/L NAA, pH
5.7。
6. a kind of floral leaf fernleaf hedge bamboo tissue culture and rapid propagation method as described in claim 1, it is characterised in that transplanted in the step 6)
Domestication specifically: it is 20000 lx that the floral leaf fernleaf hedge bamboo plant to have taken root, which is moved to intensity of illumination, and daily light application time is 16
Hour, environment temperature be 25 ± 2 DEG C, relative air humidity be 70~95% environmental condition under tame 1 week after, remove bottle cap after
Continuous training 3 days, the culture medium being attached on plant root then is washed away with 35~40 DEG C of warm water, in 1000 times of carbendazols
It impregnates 3~5 minutes, transplanting to the perlite stirred with 1000 times of carbendazols: peat: in vermiculite=1:1:1 matrix, and protecting
Hold wet, domestication terminates to can be moved to greenhouse after 1 month.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201910133402.1A CN109729976B (en) | 2019-02-22 | 2019-02-22 | Tissue culture and rapid propagation method for phoenix-tail bamboos |
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|---|---|---|---|
| CN201910133402.1A CN109729976B (en) | 2019-02-22 | 2019-02-22 | Tissue culture and rapid propagation method for phoenix-tail bamboos |
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| CN109729976B CN109729976B (en) | 2020-12-08 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113179950A (en) * | 2021-05-18 | 2021-07-30 | 广西壮族自治区农业科学院 | Method for sterilizing explants by using reduced pressure |
| CN114009342A (en) * | 2021-12-17 | 2022-02-08 | 宜宾林竹产业研究院 | Tissue culture and rapid propagation medium and tissue culture and rapid propagation method for viola tinctoria bamboo |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113179950A (en) * | 2021-05-18 | 2021-07-30 | 广西壮族自治区农业科学院 | Method for sterilizing explants by using reduced pressure |
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| CN114009342A (en) * | 2021-12-17 | 2022-02-08 | 宜宾林竹产业研究院 | Tissue culture and rapid propagation medium and tissue culture and rapid propagation method for viola tinctoria bamboo |
| CN114009342B (en) * | 2021-12-17 | 2022-07-01 | 宜宾林竹产业研究院 | Tissue culture and rapid propagation medium and tissue culture and rapid propagation method for viola tinctoria bamboo |
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