CN107047304A - A kind of tissue culture propagation method of sulphur butterfly - Google Patents
A kind of tissue culture propagation method of sulphur butterfly Download PDFInfo
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- CN107047304A CN107047304A CN201710255723.XA CN201710255723A CN107047304A CN 107047304 A CN107047304 A CN 107047304A CN 201710255723 A CN201710255723 A CN 201710255723A CN 107047304 A CN107047304 A CN 107047304A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The present invention relates to technical field of tissue culture, and in particular to a kind of tissue culture propagation method of sulphur butterfly, comprises the following steps：1) terminal bud of sulphur butterfly is taken as explant material, and explant is used to 1 ‰ mercuric chloride soaking disinfection 30min；2) explant sterilized is inoculated on MS culture mediums, the MS culture mediums include following component：6 BA, NAA, agar and sucrose, the pH of the MS culture mediums is 56；3) by step 2) MS culture mediums are put into incubator and cultivated after inoculation；The condition of culture is：26 28 degree of temperature, the 500lx of intensity of illumination 300, incubation time is 45 weeks.The beneficial effects of the present invention are：The use of terminal bud is explant, mercuric chloride disinfecting time is 30min, to improve survival rate, then obtains the optimum culture medium of sulphur butterfly by adjusting the concentration of hormone, the consumption of sucrose and pH etc. on the basis of MS culture mediums.
Description
Technical field
The present invention relates to technical field of tissue culture, and in particular to a kind of tissue culture propagation method of sulphur butterfly.
Background technology
Sulphur butterfly (Syngonium podophyllum), also known as arrow leaf taro, lac encrusted twig taro, leaf-cutting taro, fibroin rattan, fruit taro, be
Araeceae closes fruit taro platymiscium.Fruit taro strain state grace is closed, leaf changeable, color is elegant, and it is described as day south with epipremnum aureum, climing green suede
The representative Ornamental Foliage House Plants of star section.It is also that basin ornament materials is hung in current American-European interior all the fashion.
To meet the demand in market, the artificial propagation of sulphur butterfly and culture technique are by numerous studies and application, and tissue culture is numerous
It is the propagation method for being best suitable for sulphur butterfly artificial culture to grow, and tissue culture propagation technology is to realize that sulphur butterfly scale, industrialization are cultivated
With the best techniques of production.
The content of the invention
The technical problems to be solved by the invention are：It is an object of the invention to provide a kind of high sulphur butterfly of survival rate
Tissue culture propagation method.
In order to solve the above-mentioned technical problem, the technical solution adopted by the present invention is：A kind of tissue culture propagation of sulphur butterfly is provided
Method, comprises the following steps：
1) terminal bud of sulphur butterfly is taken as explant material, and explant is used to 1 ‰ mercuric chloride soaking disinfection 30min；
2) explant sterilized is inoculated on MS culture mediums, the MS culture mediums include following component：6-BA 1.5-
2mg/L, NAA 0.8-1mg/L, agar 5-7g/L and sucrose 30-40g/L, the pH of the MS culture mediums is 5-6；
3) by step 2) MS culture mediums are put into incubator and cultivated after inoculation；The condition of culture is：Temperature 26-28
Degree, intensity of illumination 300-500lx, incubation time is 4-5 weeks.
It is preferred that, in the tissue culture propagation method of above-mentioned sulphur butterfly, the MS culture mediums include following component：6-BA
1.5mg/L, NAA 0.8mg/L, agar 5g/L and sucrose 30g/L.
It is preferred that, in the tissue culture propagation method of above-mentioned sulphur butterfly, the pH of the MS culture mediums is 5.4.
It is preferred that, in the tissue culture propagation method of above-mentioned sulphur butterfly, the condition of culture is specially：26 degree of temperature, illumination
Intensity 400lx, incubation time is 4 weeks.
It is preferred that, in the tissue culture propagation method of above-mentioned sulphur butterfly, the step 1) after also include step：Explant is made
After 1 ‰ mercuric chloride soaking disinfection 25-30min, mercuric chloride erodable section is cut.
It is preferred that, in the tissue culture propagation method of above-mentioned sulphur butterfly, the step：Also wrapped after " cutting mercuric chloride erodable section "
Include step：With sterile water wash 3-5 times.
It is preferred that, in the tissue culture propagation method of above-mentioned sulphur butterfly, specifically include following steps：
1) take the terminal bud of sulphur butterfly as explant material, by explant using 1 ‰ mercuric chloride soaking disinfection 30min, cut
Mercuric chloride erodable section is removed, then with sterile water wash 5 times；
2) explant sterilized is inoculated on MS culture mediums；
3) by step 2) MS culture mediums are put into incubator and cultivated after inoculation；The condition of culture is：Temperature 26-28
Degree, intensity of illumination 300-500lx, incubation time is 4 weeks, and the MS culture mediums include following component：6-BA 1.5mg/L, NAA
0.8mg/L, agar 5g/L and sucrose 30g/L, the pH of the MS culture mediums is 5.4.
It is preferred that, in the tissue culture propagation method of above-mentioned sulphur butterfly, the step 3) after also include step：4) tissue-cultured seedling is moved
Plant, the planting matrix used during transplanting is 3 for mass ratio：1 German bog moss：Perlite.
The beneficial effects of the present invention are：The use of terminal bud is explant in the tissue culture propagation method of the sulphur butterfly of the present invention,
Mercuric chloride disinfecting time is 30min, to improve survival rate.Again on the basis of MS culture mediums by adjust hormone matched proportion density,
Consumption and pH of sucrose etc., obtain the optimum culture medium of sulphur butterfly, finally lift product quality, reduce cost.
Embodiment
To describe technology contents, the objects and the effects of the present invention in detail, it is explained below in conjunction with embodiment.
The design of most critical of the present invention is：The use of terminal bud is explant, mercuric chloride disinfecting time is 30min, to improve into
Motility rate.
Embodiment 1
A kind of tissue culture propagation method of sulphur butterfly, comprises the following steps：
1) take the terminal bud of sulphur butterfly as explant material, by explant using 1 ‰ mercuric chloride soaking disinfection 30min, cut
Mercuric chloride erodable section is removed, then with sterile water wash 3-5 times；
2) explant sterilized is inoculated on MS culture mediums；
3) by step 2) MS culture mediums are put into incubator and cultivated after inoculation；The condition of culture is：Temperature 26-28
Degree, intensity of illumination 300-500lx, incubation time is 4-5 weeks, and the MS culture mediums include following component：6-BA 1.5mg/L,
NAA 0.8mg/L, agar 5g/L and sucrose 30g/L, the pH of the MS culture mediums is 5.4.
4) tissue culture transplantation of seedlings, the planting matrix used during transplanting is 3 for mass ratio：1 German bog moss：Perlite.
Embodiment 2
A kind of tissue culture propagation method of sulphur butterfly, comprises the following steps：
1) terminal bud of sulphur butterfly is taken as explant material, and explant is used to 1 ‰ mercuric chloride soaking disinfection 30min；Cut
Remove mercuric chloride erodable section.With sterile water wash 3-5 times.
2) explant sterilized is inoculated on MS culture mediums, the MS culture mediums include following component：6-BA 2mg/
L, NAA 1mg/L, agar 7g/L and sucrose 40g/L, the pH of the MS culture mediums is 6；
3) by step 2) MS culture mediums are put into incubator and cultivated after inoculation；The condition of culture is：28 degree of temperature,
Intensity of illumination 500lx, incubation time is 5 weeks.
4) by step 3) gained tissue culture transplantation of seedlings, the planting matrix used during transplanting is 3 for mass ratio：1 German peat
Moss：Perlite.
Embodiment 3
A kind of tissue culture propagation method of sulphur butterfly, comprises the following steps：
1) take the terminal bud of sulphur butterfly as explant material, by explant using 1 ‰ mercuric chloride soaking disinfection 30min, cut
Mercuric chloride erodable section is removed, then with sterile water wash 5 times；
2) explant sterilized is inoculated on MS culture mediums, the MS culture mediums include following component：6-BA
1.5mg/L, NAA 0.8mg/L, agar 5g/L and sucrose 30g/L, the pH of the MS culture mediums is 5-6；
3) by step 2) MS culture mediums are put into incubator and cultivated after inoculation；The condition of culture is：26 degree of temperature,
Intensity of illumination 300lx, incubation time is 4 weeks.
4) by step 3) gained tissue culture transplantation of seedlings, the planting matrix used during transplanting is 3 for mass ratio：1 German peat
Moss：Perlite.
Contrast test
1st, using high about 35-40cm in resource garden, without visible scab, insect pest, no variation, no floral leaf is female with sulphur butterfly
Terminal bud and lateral bud on the healthy and strong maternal plant of this basic growth traits are used as explant.
1.1 test method
The maternal plant chosen is stopped watering to be positioned in resource garden bamboo tower, the pretreatment of 5 days is carried out.Treat its stromal surface with
Soil layer at lower 20-50mm is dried, uniformity (loose sand shape being pinched into hand, tack-free), when slight wilting is presented in plant leaf
Just its outside old leaf, dead leaf are peelled off until the compact place of terminal bud, is cut with scalpel at plant from the ground 2-3 sections.Afterwards in micro- life
On the workbench of thing room plant base portion old tissue, root eye and surrounding spot are gently cut net and bud point surrounding is cut flat with cunning, and carried out
Segmentation, per one section of bud, the explant of point good section is loaded in clean cup, is then cleaned, drained with liquid detergent.Press the big of bud
Small and old tender degree is classified, and the bud classified is positioned in aseptic empty bottle, and 8-10 are one bottle.Plant is indicated on bottle cap
Thing code and disinfecting time, are carried out disinfection with 1 ‰ mercuric chloride.Then with sterile water wash 3-5 all over (each 3-5min), cut outer
The part that implant is corroded by mercuric chloride, is inoculated on MS culture mediums (1/bottle).Specifically disinfect such as table 1.
Table 1
Condition of culture：27±1℃；300-500lx；4W, the MS culture mediums include following component：6-BA1.5mg/L,
NAA0.8mg/L, agar 5g/L and sucrose 30g/L, the pH of the MS culture mediums is 5.4.
In the tissue culture propagation method of above-mentioned sulphur butterfly, the hormone concentration of 6-BA and NAA in MS culture mediums are beneficial to white butterfly
Butterfly bud, which grows tall with blade hickie, to be occurred, and embodies breediness, effect, which is raised and lowered, in hormone concentration to decline；Sucrose concentration is
During 30g/L, sulphur butterfly hickie is substantially and more, and material growing way is sturdy, preferably, and effect, which is raised and lowered, in sucrose concentration to decline；
PH5.4 easily liquefies, beneficial to the appearance of Material growth and hickie.
2nd, result and analysis
Sulphur butterfly is after culture 4W under conditions of 27 ± 1 DEG C of temperature, intensity of illumination 300-500lx, investigation index：Bud is sprouted
Time；Very, contamination rate；The death rate；Survival rate, specific investigation result refers to table 2.
Table 2
It further investigation reveals that：This sulphur butterfly original seed makes survival rate of the survival rate (43.3%) than lateral bud of terminal bud in experiment
(21.3%) it is high, illustrate that sulphur butterfly is preferable using terminal bud as explant test effect in this experiment.In experiment handle A,
Handle in D and processing E not only without fungal contamination phenomenon but also without the phenomena of mortality, say that disinfecting time is more suitable for 30min.Handle B
The middle death rate has reached 3.9%, and reason is probably the mercuric chloride sterilization of long period, causes the death of bud.And it is dirty to handle fungi in C
Dye rate and contamination rate handle high compared with other, and reason is probably that the inadequate mercuric chloride sterilization of disinfecting time is not thoroughly caused.
Embodiments of the invention are the foregoing is only, are not intended to limit the scope of the invention, it is every to utilize this hair
The equivalents that bright description is made, or the technical field of correlation is directly or indirectly used in, similarly it is included in this hair
In bright scope of patent protection.
Claims (8)
1. a kind of tissue culture propagation method of sulphur butterfly, it is characterised in that comprise the following steps：
1) terminal bud of sulphur butterfly is taken as explant material, and explant is used to 1 ‰ mercuric chloride soaking disinfection 30min；
2) explant sterilized is inoculated on MS culture mediums, the MS culture mediums include following component：6-BA1.5-2mg/
L, NAA 0.8-1mg/L, agar 5-7g/L and sucrose 30-40g/L, the pH of the MS culture mediums is 5-6；
3) by step 2) MS culture mediums are put into incubator and cultivated after inoculation；The condition of culture is：Temperature 26-28 degree,
Intensity of illumination 300-500lx, incubation time is 4-5 weeks.
2. the tissue culture propagation method of sulphur butterfly according to claim 1, it is characterised in that the MS culture mediums include as follows
Component：6-BA 1.5mg/L, NAA 0.8mg/L, agar 5g/L and sucrose 30g/L.
3. the tissue culture propagation method of sulphur butterfly according to claim 1, it is characterised in that the pH of the MS culture mediums is
5.4。
4. the tissue culture propagation method of sulphur butterfly according to claim 1, it is characterised in that the condition of culture is specially：
26 degree of temperature, intensity of illumination 400lx, incubation time is 4 weeks.
5. the tissue culture propagation method of sulphur butterfly according to claim 1, it is characterised in that the step 1) after also include step
Suddenly：After the mercuric chloride soaking disinfection 25-30min that explant is used to 1 ‰, mercuric chloride erodable section is cut.
6. the tissue culture propagation method of sulphur butterfly according to claim 1, it is characterised in that the step：" cut mercuric chloride rotten
Also include step behind erosion part "：With sterile water wash 3-5 times.
7. the tissue culture propagation method of sulphur butterfly according to claim 1, it is characterised in that specifically include following steps：
1) take the terminal bud of sulphur butterfly as explant material, by explant using 1 ‰ mercuric chloride soaking disinfection 30min, cut liter
Mercury erodable section, then with sterile water wash 5 times；
2) explant sterilized is inoculated on MS culture mediums；
3) by step 2) MS culture mediums are put into incubator and cultivated after inoculation；The condition of culture is：Temperature 26-28 degree,
Intensity of illumination 300-500lx, incubation time is 4 weeks, and the MS culture mediums include following component：6-BA 1.5mg/L, NAA
0.8mg/L, agar 5g/L and sucrose 30g/L, the pH of the MS culture mediums is 5.4.
8. the tissue culture propagation method of sulphur butterfly according to claim 7, it is characterised in that the step 3) after also include step
Suddenly：4) by step 3) gained tissue culture transplantation of seedlings, the planting matrix used during transplanting is 3 for mass ratio：1 German bog moss：It is precious
Zhu Yan.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110627553A (en) * | 2019-10-29 | 2019-12-31 | 湖北省农业科学院经济作物研究所 | Preparation method and application of white butterfly Heguo taro still water culture solution |
| CN114145229A (en) * | 2021-09-27 | 2022-03-08 | 佛山市三水阳特园艺有限公司 | Tissue culture method for colorful Heguocao taro |
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| CN106106181A (en) * | 2016-08-08 | 2016-11-16 | 吴子平 | A kind of tissue culture and rapid propagation method of Spathiphyllum kochii |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110627553A (en) * | 2019-10-29 | 2019-12-31 | 湖北省农业科学院经济作物研究所 | Preparation method and application of white butterfly Heguo taro still water culture solution |
| CN114145229A (en) * | 2021-09-27 | 2022-03-08 | 佛山市三水阳特园艺有限公司 | Tissue culture method for colorful Heguocao taro |
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| TA01 | Transfer of patent application right |
Effective date of registration: 20180710 Address after: 352305 No. 9, tau Kou Industrial Park, Tong Kou Township, Pingnan, Ningde, Fujian Applicant after: Min Hui (Fujian) Horticulture Co., Ltd. Address before: 352300 Ling Xia Village, Ling Xia Township, Pingnan, Ningde, Fujian Applicant before: Fujian Dongjin Agriculture Technology Co. Ltd. |
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