CN104782486B - Tissue culture and intermediate propagation method for succulent Haworthia cooperivar. pilfera M. B. Bayer - Google Patents
Tissue culture and intermediate propagation method for succulent Haworthia cooperivar. pilfera M. B. Bayer Download PDFInfo
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- CN104782486B CN104782486B CN201510197424.6A CN201510197424A CN104782486B CN 104782486 B CN104782486 B CN 104782486B CN 201510197424 A CN201510197424 A CN 201510197424A CN 104782486 B CN104782486 B CN 104782486B
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Abstract
The invention discloses a tissue culture and intermediate propagation method for succulent Haworthia cooperivar. pilfera M. B. Bayer. The method comprises the following steps: selecting Haworthia cooperivar. pilfera M. B. Bayer leaves in a good growing state as explants, after disinfecting the explants, inoculating the disinfected explants to clustered shoot inducting medium for inducing clustered shoots, then placing the clustered shoots into a rooting medium for taking roots, or after subculturing of the clustered shoots, placing the clustered shoots into the rooting medium for taking roots, and then carrying out seedling adaptation and plant transplanting on intact plantlets after root growing. According to the method, Haworthia cooperivar. pilfera M. B. Bayer seedlings can be obtained by tissue culturing to Haworthia cooperivar. pilfera M. B. Bayer leaves, the material used in the method is easy to obtain, the disinfecting is convenient, the survival rate is high, the production period is short, the propagation coefficient and the propagation speed of the Haworthia cooperivar. pilfera M. B. Bayer are improved, the strain advantages of the Haworthia cooperivar. pilfera M. B. Bayer can be well kept, a great amount of excellent Haworthia cooperivar. pilfera M. B. Bayer seedlings suitable for transplanting can be propagated quickly, the production demands are met, and the economic benefits of planters are improved.
Description
Technical field
The present invention relates to a kind of plant tissue culture method for quickly breeding.
Background technology
YULU (Haworthia cooperivar.pilfera M.B.Bayer) is Folium Agaves variegatae mesh Eremurus chinensis section's (also referred to as day
Light the orchid family, Aloeaceae or A Fu flower section) volume 12 belong to the soft leaf system kind in succulent, originate in South Africa, existing
The world can cultivate on many ground.Being Dan Sheng at the beginning of plant, the most gradually in all living creatures's shape, fleshy leaf is arranged in rosette-stape, both sides
Garden is convex；Blade tip is transparent or translucent, is referred to as " window ", and there is longitudinal dark strokes on surface, and top has tiny
Must, leaf color is dark green；Loose raceme, little floral white, there is green vertical stripe.YULU is small and exquisite, blade
Glittering and translucent, as jade carving forms, peculiar and beautiful, such as lived artware, the most lovely, use
Little potted plant kind intersperses desk, desk, windowsill etc., Fresh and elegant, is famous and precious indoor appreciation flowers, is in recent years
Carrying out one of more prosperous small-sized succulent kind of popularity, by succulent, fan is pursued, and has good city
Field prospect and economic worth.
The common propagation method of YULU has cuttage, plant division, leaf to insert and the several method such as sowing.But for YULU,
These method reproduction speeds are relatively slow, inefficient, and utilize planting seed to be also unfavorable for preserving rare famous and precious product
System.And existing YULU tissue culturing system only has an example, it is to utilize YULU scape or scape ovary as outer implant
Induced bundle is sprouted, and the acquisition of material is limited by growth cycle, is unfavorable for putting into production.
Summary of the invention
The technical problem to be solved is providing the quick breeding method for tissue culture of a kind of YULU.
Solve the technical scheme that above-mentioned technical problem used to be made up of following step:
1, the selection of outer implant and sterilization
It is outer implant with YULU blade, YULU blade clean for surface clean is first divided by quality on superclean bench
Number be 0.1% mercuric chloride soak 1 minute, then soak 5 minutes with aqueous sodium hypochlorite solution that mass fraction is 5%,
All with sterilized water wash after having soaked every time, obtain aseptic explant.
2, the induction of Multiple Buds
Aseptic explant is cut into the section of 0.3～0.6cm, is inoculated in induced bundle and sprouts in culture medium,
25 ± 2 DEG C, intensity of illumination 1000～1200lux, light application time be 12～16 hours/day, relative humidity 50%～
Cultivating 14～20 days in the environment of 60%, obtain Multiple Buds, wherein induced bundle culture medium of sprouting is with MS culture medium
For minimal medium, every liter of minimal medium adds 0.8～1.2mg 6-benzyladenine, 0.8～1.2mg 6-
Glycosyl amidopurin, 25～30g sucrose, 5.0～6.0g agar, be 5.5～6.0 with NaOH regulation pH value.
3, Multiple Buds is taken root
It is after a strain separates, directly to proceed in root media root culture 3～6 weeks by Multiple Buds by 3～4 buds,
Obtain the whole plant of band root；Or it is first to proceed to subculture medium after a strain separates by Multiple Buds by 3～4 buds
Middle successive transfer culture 4～8 weeks, be then that a strain separates by the Multiple Buds after successive transfer culture by 8～10 buds, then turn
Enter in root media root culture 3～6 weeks, obtain the whole plant of band root.
Above-mentioned root culture and successive transfer culture condition are: temperature 25 ± 2 DEG C, intensity of illumination 1000～1200lux,
Light application time is 12～16 hours/day, relative humidity 50%～60%；Described root media is with 1/2MS
Culture medium is minimal medium, adds 0.08～0.12mg indolebutyric acid, 25～30g in every liter of minimal medium
Sucrose, 5.0～6.0g agar, be 5.5～6.0 with NaOH regulation pH value；Described subculture medium be with
MS culture medium is minimal medium, in every liter of minimal medium add 0.15～0.25mg 6-benzyladenine, 1～
1.5g activated carbon, 25～30g sucrose, 5.0～6.0g agar, be 5.5～6.0 with NaOH regulation pH value.
4, seedling exercising and transplanting
The whole plant of band root is opened in 25 ± 2 DEG C of incubators tissue culture bottle bottle cap, adds tap water, seedling exercising 1～
Take out after 2 days, the root media that root remains is rinsed well, In Shade air-dried, transplant to sand soil
In earth, shade, temperature be 15～25 DEG C, relative humidity be 50%～70% time growth.
In above-mentioned steps 2, preferably induced bundle culture medium of sprouting is with MS culture medium as minimal medium, every liter of base
Basal culture medium adds 1.0mg 6-benzyladenine, 1.0mg 6-glycosyl amidopurin, 25g sucrose, 5.0g fine jade
Fat, is 5.8 with NaOH regulation pH value.
In above-mentioned steps 3, preferably root media is with 1/2MS culture medium as minimal medium, every liter of basic training
Support and base adds 0.1mg indolebutyric acid, 25g sucrose, 5.0g agar, be 5.8 with NaOH regulation pH value；
Preferably subculture medium is with MS culture medium as minimal medium, adds 0.2mg 6-in every liter of minimal medium
Benzyladenine, 1g activated carbon, 25g sucrose, 5.0g agar, be 5.8 with NaOH regulation pH value.
The present invention utilizes YULU blade to be outer implant, trains by YULU leaf tissue is cultivated the group obtaining YULU
Seedling, the method material is easy to get, and sterilization is convenient, and survival rate is high, with short production cycle, improves the breeding coefficient of YULU
And reproduction speed, and can preferably keep the strain advantage of YULU, can Fast-propagation go out a large amount of be suitable for transplanting excellent
YULU seedling, meets Production requirement, improves the economic benefit of grower.
Accompanying drawing explanation
Fig. 1 is the photo of the Multiple Buds that embodiment 1 obtains.
Fig. 2 is the photo of the whole plant of the band root that embodiment 1 obtains.
Fig. 3 is the photo of the Multiple Buds obtained after embodiment 2 successive transfer culture.
Fig. 4 is the photo of the whole plant of the band root obtained after embodiment 2 root culture.
Detailed description of the invention
The present invention is described in more detail with embodiment below in conjunction with the accompanying drawings, but protection scope of the present invention not only limits
In these embodiments.
Embodiment 1
1, the selection of outer implant and sterilization
Choose YULU blade, wash YULU blade surface earth and rinse 5 hours with flowing water again, blot with filter paper
Moisture.Superclean bench first soaks YULU blade 1 minute with the mercuric chloride that mass fraction is 0.1%, then uses
Sterilized water washing 3～5 times, each 3～5 minutes；YULU blade is proceeded to the hypochlorous acid that mass fraction is 5% again
In sodium water solution soak 5 minutes, then with sterilized water wash 4～6 times, each 5～7 minutes, obtain aseptic outside
Implant.
2, the induction of Multiple Buds
Aseptic explant is cut into the section of about 0.5cm, and the induced bundle being inoculated in tissue culture bottle is sprouted in culture medium,
Every bottle graft kind 4～6,25 ± 2 DEG C, intensity of illumination 1000lux, light application time be 14 hours/day, relatively
Cultivate 14 days in the environment of humidity 50%～60%, obtain Multiple Buds.Wherein induced bundle culture medium of sprouting is with MS
Culture medium is minimal medium, adds 1.0mg 6-benzyladenine, 1.0mg 6-glycosyl in every liter of minimal medium
Amidopurin, 25g sucrose, 5.0g agar, be 5.8 with NaOH regulation pH value.As seen from Figure 1, pass through
14 days inducing culture, YULU blade induced synthesis Multiple Buds, and can differentiate on all blades 10 with
On bud.
3, Multiple Buds is taken root
It is in the root media proceeding in tissue culture bottle after a strain separates by Multiple Buds by 3～4 buds, every bottle 3～4
Strain, temperature 25 ± 2 DEG C, intensity of illumination 1000lux, light application time be 14 hours/day, relative humidity 50%～
Cultivate 3 weeks in the environment of 60%, obtain the whole plant of band root.Wherein root media is with 1/2MS culture medium
For minimal medium, every liter of minimal medium adds 0.10mg indolebutyric acid, 25g sucrose, 5.0g agar,
It is 5.8 with NaOH regulation pH value.From Figure 2 it can be seen that root culture is after 3 weeks in root media, YULU
Unrooted shoot has grown the new root of white, and physically well develops, and rooting rate reaches more than 75%.
4, seedling exercising and transplanting
The whole plant of band root is opened in 25 ± 2 DEG C of incubators tissue culture bottle bottle cap, adds tap water, seedling exercising 1
Take out after it, the root media that root remains is rinsed well, In Shade air-dried 1 day, transplant to sand
In soil, shade, temperature be 20 DEG C, relative humidity be 50%～60% time growth.
Embodiment 2
1, the selection of outer implant and sterilization
Choose YULU blade, wash YULU blade surface earth and rinse 5 hours with flowing water again, blot with filter paper
Moisture.Superclean bench first soaks YULU blade 1 minute with the mercuric chloride that mass fraction is 0.1%, then uses
Sterilized water washing 3～5 times, each 3～5 minutes；YULU blade is proceeded to the hypochlorous acid that mass fraction is 5% again
In sodium water solution soak 5 minutes, then with sterilized water wash 4～6 times, each 5～7 minutes, obtain aseptic outside
Implant.
2, the induction of Multiple Buds
Aseptic explant is cut into the section of about 0.5cm, and the induced bundle being inoculated in tissue culture bottle is sprouted in culture medium,
Every bottle graft kind 4～6,25 ± 2 DEG C, intensity of illumination 1000lux, light application time be 14 hours/day, relatively
Cultivate 14 days in the environment of humidity 50%～60%, obtain Multiple Buds.Wherein induced bundle culture medium of sprouting is with MS
Culture medium is minimal medium, adds 1.0mg 6-benzyladenine, 1.0mg 6-glycosyl in every liter of minimal medium
Amidopurin, 25g sucrose, 5.0g agar, be 5.8 with NaOH regulation pH value.
3, Multiple Buds is taken root
It is in the subculture medium proceeding in tissue culture bottle after a strain separates by Multiple Buds by 3～4 buds, every bottle 3～4
Strain, is 14 hours/day, relative humidity 50%-60% in temperature 25 ± 2 DEG C, intensity of illumination 1000lux, light application time
In the environment of cultivate 4 weeks, be then a strain separation by the Multiple Buds after successive transfer culture by 8～10 buds, then proceed to
In root media in tissue culture bottle, every bottle 3～4 strains, at temperature 25 ± 2 DEG C, intensity of illumination 1000lux, light
It is to cultivate 3 weeks in the environment of 14 hours/day, relative humidity 50%～60% according to the time, obtains completely planting of band root
Strain.Wherein root media is with 1/2MS culture medium as minimal medium, adds in every liter of minimal medium
0.10mg indolebutyric acid, 25g sucrose, 5.0g agar, be 5.8 with NaOH regulation pH value.As seen from Figure 3,
Successive transfer culture formed after 4 weeks grow fine, healthy and strong Multiple Buds bunch；From fig. 4, it can be seen that entered again by after successive transfer culture
The seedling of row root culture takes root good, sturdy.
4, seedling exercising and transplanting
The whole plant of band root is opened in 25 ± 2 DEG C of incubators tissue culture bottle bottle cap, adds tap water, seedling exercising 1
Take out after it, the root media that root remains is rinsed well, In Shade air-dried 1 day, transplant to sand
In soil, shade, temperature be 20 DEG C, relative humidity be 50%～60% time growth.
Claims (4)
1. the quick breeding method for tissue culture of a succulent YULU, it is characterised in that it comprises the steps:
(1) selection of outer implant and sterilization
It is outer implant with YULU blade, YULU blade clean for surface clean is first divided by quality on superclean bench
Number be 0.1% mercuric chloride soak 1 minute, then soak 5 minutes with aqueous sodium hypochlorite solution that mass fraction is 5%,
All with sterilized water wash after having soaked every time, obtain aseptic explant；
(2) induction of Multiple Buds
Aseptic explant is cut into the section of 0.3～0.6cm, is inoculated in induced bundle and sprouts in culture medium,
25 ± 2 DEG C, intensity of illumination 1000～1200lux, light application time be 12～16 hours/day, relative humidity 50%～
Cultivating 14～20 days in the environment of 60%, obtain Multiple Buds, wherein induced bundle culture medium of sprouting is with MS culture medium
For minimal medium, every liter of minimal medium adds 0.8～1.2mg 6-benzyladenine, 0.8～1.2mg 6-
Glycosyl amidopurin, 25～30g sucrose, 5.0～6.0g agar, be 5.5～6.0 with NaOH regulation pH value；
(3) Multiple Buds is taken root
It is after a strain separates, directly to proceed in root media root culture 3～6 weeks by Multiple Buds by 3～4 buds,
Obtain the whole plant of band root；Or it is first to proceed to subculture medium after a strain separates by Multiple Buds by 3～4 buds
Middle successive transfer culture 4～8 weeks, be then that a strain separates by the Multiple Buds after successive transfer culture by 8～10 buds, then turn
Enter in root media root culture 3～6 weeks, obtain the whole plant of band root；
Above-mentioned root culture and successive transfer culture condition are: temperature 25 ± 2 DEG C, intensity of illumination 1000～1200lux,
Light application time is 12～16 hours/day, relative humidity 50%～60%；Described root media is with 1/2MS
Culture medium is minimal medium, adds 0.08～0.12mg indolebutyric acid, 25～30g in every liter of minimal medium
Sucrose, 5.0～6.0g agar, be 5.5～6.0 with NaOH regulation pH value；Described subculture medium be with
MS culture medium is minimal medium, in every liter of minimal medium add 0.15～0.25mg 6-benzyladenine, 1～
1.5g activated carbon, 25～30g sucrose, 5.0～6.0g agar, be 5.5～6.0 with NaOH regulation pH value；
(4) seedling exercising and transplanting
The whole plant of band root is opened in 25 ± 2 DEG C of incubators tissue culture bottle bottle cap, adds tap water, seedling exercising 1～
Take out after 2 days, the root media that root remains is rinsed well, In Shade air-dried, transplant to sand soil
In earth, shade, temperature be 15～25 DEG C, relative humidity be 50%～70% time growth.
The quick breeding method for tissue culture of succulent YULU the most according to claim 1, its feature exists
In: in described step (2), induced bundle culture medium of sprouting is with MS culture medium as minimal medium, every liter
Minimal medium adds 1.0mg 6-benzyladenine, 1.0mg 6-glycosyl amidopurin, 25g sucrose, 5.0g
Agar, is 5.8 with NaOH regulation pH value.
The quick breeding method for tissue culture of succulent YULU the most according to claim 1, its feature exists
In: in described step (3), root media is with 1/2MS culture medium as minimal medium, and every liter is basic
Culture medium adds 0.1mg indolebutyric acid, 25g sucrose, 5.0g agar, is 5.8 with NaOH regulation pH value.
The quick breeding method for tissue culture of succulent YULU the most according to claim 1, its feature exists
In: in described step (3), subculture medium is with MS culture medium as minimal medium, every liter of basic training
Support and base adds 0.2mg 6-benzyladenine, 1g activated carbon, 25g sucrose, 5.0g agar, adjust with NaOH
Joint pH value is 5.8.
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| CN105010147A (en) * | 2015-08-14 | 2015-11-04 | 泓柯（天津）农业科技有限公司 | Special culture medium for improving tissue culture propagation speed of haworthia succulent plants and tissue culture method |
| CN105230487A (en) * | 2015-10-13 | 2016-01-13 | 宁波市鄞州万众生态果园专业合作社 | Haworthia cooperivar. pilfera M. B. Bayer OB-1 tissue culture formula |
| CN105638463A (en) * | 2015-12-30 | 2016-06-08 | 四川禾木本业农林科技有限公司 | Tissue-culture rapid propagation method for succulents |
| CN105766635B (en) * | 2016-03-14 | 2017-12-22 | 龙岩市禾康生物科技有限公司 | A kind of method of succulent tissue-culturing rapid propagation |
| CN106258397A (en) * | 2016-08-12 | 2017-01-04 | 广西鑫雅皇庭园林工程有限责任公司 | A kind of implantation methods of Bryophyllum succulent |
| CN106258902A (en) * | 2016-08-31 | 2017-01-04 | 昆明理工大学 | The efficient leaf of a kind of succulent inserts propagation method |
| CN106857256B (en) * | 2017-02-21 | 2019-03-12 | 淮北师范大学 | The method for improving beautiful dew breeding potential based on callus induction Regeneration Ways |
| CN107155890B (en) * | 2017-06-08 | 2021-02-09 | 北京农学院 | Jade dew in-vitro rapid propagation method taking leaves as explants |
| CN107278849B (en) * | 2017-06-29 | 2021-02-09 | 安徽强农牧业有限公司 | Preparation method of breathable rooted succulent plant culture medium particles |
| CN108719065A (en) * | 2018-05-21 | 2018-11-02 | 句容市茂润苗木有限公司 | A kind of succulent rapid propagation method |
| CN108401911A (en) * | 2018-06-13 | 2018-08-17 | 内蒙古自治区生物技术研究院 | Jade dew tissue culture medium (TCM) and preparation method thereof |
| CN110089431A (en) * | 2019-05-06 | 2019-08-06 | 绵阳师范学院 | The quick breeding by group culture method of jade dew |
| CN110447538B (en) * | 2019-08-30 | 2022-05-10 | 江苏省中国科学院植物研究所 | Tissue culture method taking pennisetum panzeri inflorescence axes as explants |
| CN115399243A (en) * | 2022-08-26 | 2022-11-29 | 内蒙古科技大学 | Tissue culture rapid propagation method for inducing regeneration of adventitious buds by taking rubiaceae yulu leaves as explants |
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| CN102499090B (en) * | 2011-11-07 | 2013-07-24 | 上海旭东园艺有限公司 | Method for isolated culture of Haworthia succulent plants |
| CN103141387A (en) * | 2013-03-08 | 2013-06-12 | 浙江省农业科学院 | Method for cultivating haworthia maughanii tissue |
| CN105230487A (en) * | 2015-10-13 | 2016-01-13 | 宁波市鄞州万众生态果园专业合作社 | Haworthia cooperivar. pilfera M. B. Bayer OB-1 tissue culture formula |
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