CN109479717A - A kind of method of Pearl color osmanthus explant disinfection - Google Patents
A kind of method of Pearl color osmanthus explant disinfection Download PDFInfo
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- CN109479717A CN109479717A CN201811593818.3A CN201811593818A CN109479717A CN 109479717 A CN109479717 A CN 109479717A CN 201811593818 A CN201811593818 A CN 201811593818A CN 109479717 A CN109479717 A CN 109479717A
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- explant
- pearl color
- color osmanthus
- osmanthus
- disinfection
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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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- Developmental Biology & Embryology (AREA)
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Abstract
The invention discloses a kind of methods of Pearl color osmanthus explant disinfection, including the selection of Pearl color osmanthus explant material, the processing of Pearl color osmanthus explant, Pearl color osmanthus explant culture.Optimization process method of the invention is that the HgCl that 2 min, aseptic water washing 3 times, ethanol postincubation 35-50 s of 75wt%, aseptic water washing 3 times, 0.1wt% are impregnated in the liquor natrii hypochloritis of 30 min, 2 wt% is rinsed in flowing water2Sterilize the moisture that 8 min, aseptic water washing 4-5 suck explant surface all over, aseptic filter paper.The method of the present invention is simple and easy, and after the disinfection of Pearl color osmanthus explant, pollution rate is reduced to 16%, and survival rate significantly improves.
Description
Technical field
The present invention relates to a kind of methods of Pearl color osmanthus explant disinfection.
Background technique
Pearl color osmanthus is the mutation of wooden slippers category sweet osmanthus, and leaf is oval sawtooth, and blade is compared to common greenery sweet osmanthus
It is thicker.Leaf can be with perenniaity turn colors.Early spring tender shoots be purplish red rhodo, after switch to it is pink, and then turn golden yellow
To white.Started again from central main lobe arteries and veins to both sides gradual change green.Until dark green, one Nian Si under conditions of weather conditions are suitable for
Season constantly has tender shoots to emerge, therefore entire plant pearl is of a riot of colour from top to bottom, deep purples and bright reds-beautiful flowersa riot of brilliant purple and tender crimson.
Currently, using the more of one or both disinfectant during the disinfection of plant explant.Use single disinfection method
Agent processing is difficult to obtain sterilizable material；It is handled using two kinds of disinfectants, aseptic explant can be obtained, but obtain sterilizable material
Sterile rate is not high, and during the tissue culture in later period, it may appear that different degrees of germ contamination, or even in culture 7-10d mould
Pollution rate increases, and causes sterile rate lower；It, not only can be with by the present invention in that with three kinds of disinfectant Combined Treatment explant materials
Achieve the effect that thorough disinfection, and higher sterile rate can be obtained.Outside using three kinds of disinfectant Combined Treatment Pearl color osmanthus
Implant, can capturing material at any time at all seasons, carry out disinfection processing, and can get higher sterilizable material.
Summary of the invention
The purpose of the present invention is to provide a kind of methods of Pearl color osmanthus explant disinfection.
To achieve the above object, the present invention adopts the following technical scheme:
A kind of method of Pearl color osmanthus explant disinfection, comprising the following steps:
(1) Pearl color osmanthus explant material selects: choosing bud from stalwartness, no disease and pests harm, well-grown Pearl color osmanthus maternal plant
For full, annual, semi-lignified the branch of point as explant material, moisturizing processing is spare；
(2) Pearl color osmanthus explant is handled: the branch of step (1) being cut into 0.5 cm-1.5 cm segment, removes blade and leaf
Handle rinses 30 min in flowing water, with soft brush wash explant surface, after being handled with NaClO solution, then at alcohol
Reason, finally uses HgCl2It sterilizes, sucks the moisture on explant surface after aseptic water washing 4-5 times with aseptic filter paper, be packed into sterile
It is spare in bottle；Pollution rate and survival rate are counted after 30 d；
(3) Pearl color osmanthus explant condition of culture: 25 ± 3 DEG C of cultivation temperature, 12 h d of light application time-1, intensity of illumination 3500
Lux。
The NaClO solution processing methods are as follows: impregnate 2 min with the NaClO solution of 2wt%.
The ethanol postincubation method are as follows: with the 35 s-50 s of ethanol postincubation of 75wt%.
The HgCl2Sterilization method are as follows: with the HgCl of 0.1wt%2Sterilize 5- 11min.
It carries out disinfection place the present invention has the advantages that will use one or both disinfectant generally directed to explant disinfection
Reason, by the present invention in that not only can achieve the effect of thorough disinfection with three kinds of disinfectant Combined Treatment explant materials, but also
Higher sterile rate can be obtained.It, can at all seasons at any time using three kinds of disinfectant Combined Treatment Pearl color osmanthus explants
Capturing material, having broken only can get higher in the limitation that spring acquisition explant can get sterilizable material by the method
Sterilizable material is provided fundamental basis for industrial seedling rearing.It is carried out disinfection using sterilization method of the present invention to Pearl color osmanthus explant
Afterwards, explant pollution rate drops to 16%, and survival rate significantly improves, and can reach 73.3%, and significant effect of the present invention is simple and easy,
Technical support is provided for Pearl color osmanthus the factorial production.
Specific embodiment
Below will by example, the invention will be further described, these descriptions are not to make to the content of present invention into one
The restriction of step.It should be understood by those skilled in the art that equivalent replacement made by the content of present invention, or be correspondingly improved, still belong to
Within protection scope of the present invention.
Embodiment 1
1. test material
The new woods sweet osmanthus nursery stock in material to be tested source Guilin Lipu.It is selected from stalwartness, no disease and pests harm, well-grown maternal plant
Take full, annual, semi-lignified the branch of bud point as explant material, tissue culture room is taken back in moisturizing processing.
2. test method
Branch is cut into 0.5 cm -1.5 cm, removes blade and petiole, 30 min is rinsed in flowing water, with soft toothbrush
Explant surface is cleaned, 2 min, then 35 s of ethanol postincubation and 50 s two with 75wt% are impregnated in the NaClO solution of 2wt%
Gradient finally uses the HgCl of 0.1wt%2Sterilize 5,8 and 11min(and be grouped processing by the processing method in table 1), sterile water
The moisture for sucking explant surface after rinsing 4-5 times with aseptic filter paper, is fitted into spare in aseptic bottle.Each disinfection treatment is three times
It repeats, each 85 stem sections, is inoculated with 1530 stem sections altogether.Compare the sterilization effect of different disinfecting times, counts pollution rate after 30 d
And survival rate.Condition of culture: 25 ± 3 DEG C of cultivation temperature, 12 h d of light application time-1, 3500 Lux of intensity of illumination.
3. test result
From table 1 it follows that explant disinfection in Pearl color osmanthus uses the alcohol and 0.1wt%HgCl of 75wt%2Combined treatment, with
The extension of disinfecting time, pollution rate it is gradually lower.0.1wt%HgCl2When sterilization time 5min, the pollution of processing 1 and processing 4
Rate is above 20%.Use the alcohol disinfecting 35s or 50s, 0.1wt%HgCl of 75wt%28min is sterilized, pollution rate is 16% He respectively
17%, survival rate is 68.3% and 73.3%, the two processing pollution rates are lower, and survival rate is higher, and without significant between the two processing
Difference, the method that all can serve as the disinfection of Pearl color osmanthus explant.
Influence of the different sterilization treatments of table 1 to explant sterilization effect
Note: data are mean+SD in table；Same column difference letter represent significant difference (P< 0.05).
According to above-mentioned analysis, the optimal disinfection treatment method in Pearl color osmanthus are as follows: the secondary chlorine of 30 min, 2wt% is rinsed in flowing water
Impregnated in acid sodium solution 2 min, aseptic water washing 3 times, 75 % ethanol postincubation 35 s or 50 s, aseptic water washing 3 times,
The HgCl of 0.1wt%2Sterilize the moisture that 8 min, aseptic water washing 4-5 suck explant surface all over, aseptic filter paper.
The foregoing is merely presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, is all covered by the present invention.
Claims (4)
1. a kind of method of Pearl color osmanthus explant disinfection, which comprises the following steps:
(1) Pearl color osmanthus explant material selects: choosing bud from stalwartness, no disease and pests harm, well-grown Pearl color osmanthus maternal plant
For full, annual, semi-lignified the branch of point as explant material, moisturizing processing is spare；
(2) Pearl color osmanthus explant is handled: the branch of step (1) being cut into 0.5 cm-1.5 cm segment, removes blade and leaf
Handle rinses 30 min in flowing water, with soft brush wash explant surface, after being handled with NaClO solution, then at alcohol
Reason, finally uses HgCl2It sterilizes, sucks the moisture on explant surface after aseptic water washing 4-5 times with aseptic filter paper, be packed into sterile
It is spare in bottle；Pollution rate and survival rate are counted after 30 d；
(3) Pearl color osmanthus explant condition of culture: 25 ± 3 DEG C of cultivation temperature, 12 h d of light application time-1, intensity of illumination 3500
Lux。
2. the method for a kind of Pearl color osmanthus explant disinfection according to claim 1, which is characterized in that the NaClO solution
Processing method are as follows: impregnate 2 min with the NaClO solution of 2wt%.
3. the method for a kind of Pearl color osmanthus explant disinfection according to claim 1, which is characterized in that the ethanol postincubation side
Method are as follows: with the 35 s-50 s of ethanol postincubation of 75wt%.
4. the method for a kind of Pearl color osmanthus explant disinfection according to claim 1, which is characterized in that the HgCl2Disinfection side
Method are as follows: with the HgCl of 0.1wt%2Sterilize 5- 11min.
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| CN201811593818.3A CN109479717A (en) | 2018-12-25 | 2018-12-25 | A kind of method of Pearl color osmanthus explant disinfection |
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| CN201811593818.3A CN109479717A (en) | 2018-12-25 | 2018-12-25 | A kind of method of Pearl color osmanthus explant disinfection |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102246700A (en) * | 2011-08-18 | 2011-11-23 | 西南林业大学 | Tissue culture method for populus yunnanensis Dode with tender stem as explant |
| CN104365477A (en) * | 2014-09-12 | 2015-02-25 | 上海绿力农业科技有限公司 | Secondary proliferation culture method of color osmanthusfragrans test-tube plantlets |
| CN106613979A (en) * | 2016-12-23 | 2017-05-10 | 江苏农林职业技术学院 | Primary culture method of cupressus macrocarpa goldcres |
| CN106900548A (en) * | 2017-03-28 | 2017-06-30 | 河南红枫种苗股份有限公司 | A kind of breeding method of descendants of royal families coloured silk leaf kind |
| CN108308030A (en) * | 2018-04-04 | 2018-07-24 | 中国热带农业科学院南亚热带作物研究所 | A method of promoting avocado stem section evoked callus and sterile bud |
-
2018
- 2018-12-25 CN CN201811593818.3A patent/CN109479717A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102246700A (en) * | 2011-08-18 | 2011-11-23 | 西南林业大学 | Tissue culture method for populus yunnanensis Dode with tender stem as explant |
| CN104365477A (en) * | 2014-09-12 | 2015-02-25 | 上海绿力农业科技有限公司 | Secondary proliferation culture method of color osmanthusfragrans test-tube plantlets |
| CN106613979A (en) * | 2016-12-23 | 2017-05-10 | 江苏农林职业技术学院 | Primary culture method of cupressus macrocarpa goldcres |
| CN106900548A (en) * | 2017-03-28 | 2017-06-30 | 河南红枫种苗股份有限公司 | A kind of breeding method of descendants of royal families coloured silk leaf kind |
| CN108308030A (en) * | 2018-04-04 | 2018-07-24 | 中国热带农业科学院南亚热带作物研究所 | A method of promoting avocado stem section evoked callus and sterile bud |
Non-Patent Citations (1)
| Title |
|---|
| 吕志鹏: "金桂的组织培养与快速繁殖", 《安徽农学通报》 * |
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