CN107151644A - A kind of plant leaf blade conservation in vitro method and its application - Google Patents
A kind of plant leaf blade conservation in vitro method and its application Download PDFInfo
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- CN107151644A CN107151644A CN201710312053.0A CN201710312053A CN107151644A CN 107151644 A CN107151644 A CN 107151644A CN 201710312053 A CN201710312053 A CN 201710312053A CN 107151644 A CN107151644 A CN 107151644A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/04—Plant cells or tissues
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0226—Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/999—Small molecules not provided for elsewhere
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Abstract
The invention discloses a kind of plant leaf blade conservation in vitro method, it is related to technical field of plant culture.The inventive method includes：Choose whole leaf and leave and take petiole as preculture leaf；The laying culture paper on the inner surface of glass dish bottom, nutrient solution is poured on culture paper；The front of preculture leaf is placed with the culture paper surface relative close containing nutrient solution, the petiole part of preculture leaf is close to culture paper to absorb the nutrient solution in culture paper；With preservative film seal glass ware side wall；Seal glass ware is placed in culturing room and cultivated；The thermostat temperature of culturing room is 25 DEG C 28 DEG C, is 100 days 25 days per the life time-to-live of the 10h 14h preculture leaves alternately selected in dark or photoenvironment culture, seal glass ware.The inventive method simple and quick can carry out cultured in vitro to blade, and the blade of cultured in vitro is completely pollution-free, and blade life can be maintained 100 days 25 days.
Description
Technical field
The present invention relates to technical field of plant culture, more particularly to a kind of plant leaf blade conservation in vitro method and its should
With.
Background technology
Plant leaf blade Plantlet in vitro can provide fresh and alive study sample for field scientific research, be usually used in the life of plant leaf blade
Reason measure, genetic test and pathological research.
Poplar Species are various, strong adaptability, and growth is fast, is the preferred commerical tree species in countries in the world and the energy, Woody feed
Seeds.The introducing a fine variety of willow, hybridize and Extend culture is one of important content of China's forestry development.The collection of field sample, be
Abundant Poplar Varieties, the necessary links of scientific research；Traditional field acquisition is limited, it is necessary to which sample is put because of time, temperature etc.
Taken back in the Cryo Equipments such as curling stone, have the shortcomings that carrying is inconvenient, cost is high, the device space is big, the sample time-to-live is short.
In the physiological measurement, genetic test and pathological research of willow, collection poplar leaf and progress desk research
Also the problem of facing same；Leaf disk method has been invented for this G.P.Clinton etc. (1924), has been beaten with card punch and takes leaf cake, 50%
Rinse 2-3 times, then be placed in above 1-1.5% solid agar medium after alcohol surface sterilization, illumination is protected after sealing culture dish
Deposit, this method has inoculation strain purity height, inoculum density quantification, temperature and illumination etc. when carrying out physiology, pathological study
The advantages of experiment condition is easy to control, statistics is convenient, easily repetition and field investigation uniformity are good, but blade loss is big, naturally downright bad
Rate is high, easily contaminated, leaf dish short life, inconvenient operation the shortcomings of.
Then, Littlefield L.J. (1981), Hitoshi Nakamura (1998) etc. are successively carried out to this method
Improve, domestic scholars Hu Jingjiang (2000), Tian Chengming (2002), Du Lin (2005) etc. are also successively changed to this method
Enter, but necrosis and pollution are still the serious problems faced in research.
The content of the invention
In view of this, the embodiments of the invention provide a kind of plant leaf blade conservation in vitro method and its application, mainly
Short technical problem that purpose is that blade loss is big when solving leaf culture in vitro, natural necrosis rate is high and life is held time.
To reach above-mentioned purpose, invention broadly provides following technical scheme：
On the one hand, the embodiments of the invention provide a kind of plant leaf blade conservation in vitro method, methods described is included such as
Lower step：Choose whole leaf and leave and take petiole as preculture leaf；
The laying culture paper on the inner surface of glass dish bottom, nutrient solution is poured on the culture paper；
The front of the preculture leaf is placed with the culture paper surface relative close containing the nutrient solution, made described pre-
It is close to the culture paper to absorb the nutrient solution in the culture paper in the petiole part of culture leaf；
The glass dish formation seal glass ware is sealed with preservative film, the seal glass ware is placed in culturing room and carried out
Culture；Wherein, the thermostat temperature of the culturing room is 25 DEG C -28 DEG C, per 10h-14h alternately from the training of dark or photoenvironment
Support, the life time-to-live of the preculture leaf in the seal glass ware is -100 days 25 days.
Preferably, the plant leaf blade is poplar leaf；The petiole length left and taken is 2cm-3cm, leaves and takes the mesh of petiole
The blade that is to aid in carry out water balance to maintain blade vitality；The willow is preferably Cathay poplar, black poplar and its hybrid poplar
Blade.
Preferably, the area of the culture paper is 1 to two/3rd point of the inner surface area of the glass dish bottom
One of, it is therefore an objective to the face of blade in glass dish is seen light；The culture paper can extend to the side wall of the glass dish.
Preferably, the culture paper is neutral filter paper, normal domestic use toilet paper or general newspaper；Culture dish is common
Glass culture dish or batch cultur ware, preservative film or sealed membrane can be employed as the encapsulant of culture dish side wall.
Preferably, the nutrient solution is the gibberellin aqueous solution, ordinary tap water or sterilized water；Fallen on to the culture paper
Enter the nutrient solution and refer to that the culture paper is all soaked and flowed out without surplus liquid by the nutrient solution, i.e., described culture paper is inhaled
Receive and saturation state is basically reached after the nutrient solution to maintain in the glass dish Rh as 100%.
Preferably, the volumetric concentration of the gibberellin aqueous solution is ‰ g/l of 1-10.
Preferably, the photoenvironment is to use lighting, the lighting is to use 5-8 roots power for 15 watts
LED lamp tube；The fluorescent tube brand is Ou Pu fluorescent tubes.
Preferably, relative humidity in the seal glass ware for 100% so that blade sustains life power, stomata
Open, effectively carry out photosynthesis；The life time-to-live of the preculture leaf in the seal glass ware is -60 days 40 days.
Preferably, staggered relatively refer to tips upside down on the preculture leaf on the culture paper, the preculture leaf
Front refer to the one side of the blade that those skilled in the art unanimously accept, the i.e. bright-coloured one side of color light, with vacuum side of blade
Relatively；By the blade back-off in it is described culture paper on after, most handy tweezers carefully adjust petiole height so that the petiole and
Filter paper is fully contacted.
On the other hand, surveyed the embodiments of the invention provide above-mentioned culture store method in Plant seed preserving, plant genetic
Application in fixed, physiologically active of plant measure and Pathogenicity.
Compared with prior art, the beneficial effects of the invention are as follows：
The present invention preserves that the natural necrosis rate that plant leaf blade occurs is high, blade loss is big and blade for leaf disk method culture
Life is held time short technical problem, and employing will stay petiolate whole leaf to be put in the glass dish of closing, the glass
Ware bottom is equipped with culture paper, contains nutrient solution in the culture paper, and the petiole can be in close contact culture paper, and by above-mentioned closing
Glass dish be put into 25 DEG C of -28 DEG C of isoperibols and the technology alternately cultivated per 10h-14h from dark or photoenvironment
Means, have reached that the blade by culture preservation is complete, blade is pollution-free, life can maintain -100 days 25 days and simple to operate
Quick technical purpose, is determined, physiologically active of plant is determined and Pathogenicity provides for Plant seed preserving, plant genetic
Preferable store method.
Embodiment
For further illustrate the present invention to reach the technological means and effect that predetermined goal of the invention is taken, below with compared with
Good embodiment, to embodiment, technical scheme, feature and its effect according to the present patent application, is described in detail as after.Under
State it is bright in multiple embodiments in special characteristic, structure or feature can be combined by any suitable form.
Embodiment 1
The full wafer spire of any leaf age health of 69 poplars is chosen, the petiole of 2-3cm length is retained, prepares clean glass culture dish
(Φ=90), clean neutral filter paper, the neutral filter paper of clip glass culture dish area 1/2 is layed in as culture paper
Culture dish bottom, above-mentioned neutral filter paper can be protruded along ware wall, configuration extra fine quality than the gibberellin aqueous solution (GA3It is water-soluble
Liquid), measure the above-mentioned gibberellin aqueous solution of 3mL and be poured into above-mentioned neutral filter paper, and make above-mentioned neutral filter paper completely moistening and
Without surplus liquid outflow；
By above-mentioned preculture leaf back-off in above-mentioned neutral its surface, make preculture blade base (i.e. petiolate part)
Closely it is affixed on filter paper, the remainder of preculture leaf can be on neutral filter paper, and petiole is carefully adjusted with tweezers highly makes leaf
Handle and neutral filter paper are fully contacted；Above-mentioned glass culture dish is sealed with preservative film, laboratory is taken back standby；
Above-mentioned glass culture dish is put into 26 DEG C of constant temperature, dark or photoenvironment is used alternatingly within every 12 hours and is cultivated, 60
After it, above-mentioned 69 poplar blade is still bud green and can sprout complete root system at petiole, and 90 days rear blades still have stronger
Vitality.
Embodiment 2
The selection too white poplar whole leaf of 6-10 days, the petiole of retention 2-3cm length, the clean glass culture dish of preparation (Φ=
90), clean neutral filter paper, the neutral filter paper of clip glass culture dish area 1/2 is layed in culture dish as culture paper
Bottom, above-mentioned neutral filter paper can be protruded along ware wall, and configuration concentration is 10 ‰ g/l of the gibberellin aqueous solution (GA3The aqueous solution),
Measure the above-mentioned gibberellin aqueous solution of 3mL and be poured into above-mentioned neutral filter paper, and make above-mentioned neutral filter paper completely moistening and without many
Extraction raffinate body flows out；Two treatment groups are set, treatment group 1, treatment group 2 are named respectively；Each treatment group handles 4 leaves, totally 8
Blade.
After being ground rapidly to above-mentioned too white poplar excised leaf using liquid nitrogen, save backup, carried using 95% ethanol in -20 DEG C
The chlorophyll content that (Wang Yuan armies 2010) determines the too white poplar excised leaf that above-mentioned liquid nitrogen grinding is preserved is followed the example of, crude enzyme liquid is prepared simultaneously
It is placed on 4 DEG C of preservations；
Physiological index determining (Hodges et al.1999 are carried out to above-mentioned crude enzyme liquid；Hao again it is refined 2006)：Using coomassie
Light blue G-250 methods determine the content of soluble protein of above-mentioned crude enzyme liquid, determine above-mentioned using nitroblue tetrazolium (NBT) photoreduction met hod
Superoxide dismutase (SOD) activity of crude enzyme liquid, determines above-mentioned peroxidase (POD) activity using guaiacol method, surveys
Determine result as shown in table 1, from measurement result, the excised leaf of above-mentioned too white poplar can maintain each physical signs basically identical.
The too white poplar leaf culture in vitro physical signs difference of table 1.
Embodiment 3
The too white poplar healthy 5 group one-piece blade of 5-10 days is chosen, the petiole of 2-3cm length is retained, prepares clean glass training
Ware (Φ=90), clean neutral filter paper are supported, the neutral filter paper of clip glass culture dish area 1/2 is used as culture paper, Jiang Qipu
Located at culture dish bottom, above-mentioned neutral filter paper can be protruded along ware wall, and configuration concentration is 10 ‰ g/l of the gibberellin aqueous solution
(GA3The aqueous solution), measure the above-mentioned gibberellin aqueous solution of 3mL and be poured into above-mentioned neutral filter paper, and make above-mentioned neutral filter paper complete
Moisten and flowed out without surplus liquid entirely；
By above-mentioned preculture leaf back-off in above-mentioned neutral its surface, make preculture blade base (i.e. petiolate part)
Closely it is affixed on filter paper, petiole is carefully adjusted with tweezers highly makes petiole and neutral filter paper fully contact；Will be above-mentioned with preservative film
Glass culture dish is sealed, and takes back laboratory standby；
Loose poplar grid rest fungus (Melampsora larici-populina Kleb.) uredospore is made 1 × 105It is sterile
Aqueous suspensions, make liquid not flow with the careful too white poplar blade back that is sprayed on suspension of watering can；
Culture dish is sealed with preservative film, in 26 DEG C of constant temperature, 12 hours dark/illumination alternate cultures；Observation and record incobation
Phase, uredospore breeding situation；As a result show, 5 tissue cultures in this method support blade, obtain consistent experimental result, incubation period 6
My god, uredium yield big, no living contaminants more can continuously harvest uredospore, blade is without the obvious phenomena of mortality in 20 days.
Embodiment 4
The present embodiment 4 and the difference of embodiment 3 are that culture paper uses general newspaper, and too white poplar is in vitro after 40 days
Blade remains to produce uredium, but mesophyll tissue has started to necrosis.
From the test result of the embodiment 4 of above example 1, it can be maintained using the method for cultured in vitro blade of the present invention
Blade vitality -100 days 25 days；It is complete, pollution-free using the blade of the inventive method culture, every physical signs base of blade
This stabilization；The inventive method is simple to operate quick；The inventive method is Plant seed preserving, plant genetic measure, plant physiology
Determination of activity and Pathogenicity provide preferable store method.
Not most part in the embodiment of the present invention, those skilled in the art can select from the prior art.
Disclosed above is only the embodiment of the present invention, but protection scope of the present invention is not limited thereto, and is appointed
What those familiar with the art the invention discloses technical scope in, change or replacement can be readily occurred in, all should
It is included within the scope of the present invention.Therefore, protection scope of the present invention should using above-mentioned scope of the claims as
It is accurate.
Claims (9)
1. a kind of plant leaf blade conservation in vitro method, it is characterised in that methods described comprises the following steps：
Choose whole leaf and leave and take petiole as preculture leaf；
The laying culture paper on the inner surface of glass dish bottom, nutrient solution is poured on the culture paper；
The front of the preculture leaf is placed with the culture paper surface relative close containing the nutrient solution, makes the preculture
It is close to the culture paper to absorb the nutrient solution in the culture paper in the petiole part of leaf；
The glass dish formation seal glass ware is sealed with preservative film, the seal glass ware is placed in culturing room and trained
Support；Wherein, the thermostat temperature of the culturing room is 25 DEG C -28 DEG C, per 10h-14h alternately from dark or photoenvironment culture,
The life time-to-live of the preculture leaf in the seal glass ware is -100 days 25 days.
2. a kind of plant leaf blade conservation in vitro method according to claim 1, it is characterised in that the plant leaf blade
For poplar leaf；The petiole length left and taken is 2cm-3cm.
3. a kind of plant leaf blade conservation in vitro method according to claim 1, it is characterised in that the culture paper
Area is 1st/1st to two/3rd of the inner surface area of the glass dish bottom.
4. a kind of plant leaf blade conservation in vitro method according to claim 1, it is characterised in that the culture paper is
Neutral filter paper, normal domestic use toilet paper or general newspaper.
5. a kind of plant leaf blade conservation in vitro method according to claim 1, it is characterised in that the nutrient solution is
The gibberellin aqueous solution, ordinary tap water or sterilized water.
6. a kind of plant leaf blade conservation in vitro method according to claim 5, it is characterised in that the gibberellin water
The volumetric concentration of solution is ‰ g/l of 1-10.
7. a kind of plant leaf blade conservation in vitro method according to claim 1, it is characterised in that the photoenvironment
It is to use lighting, the lighting is to use 5-8 roots power for 15 watts of LED lamp tube.
8. a kind of plant leaf blade conservation in vitro method according to claim 1, it is characterised in that the seal glass
Relative humidity in ware is 100%；The life time-to-live of the preculture leaf in the seal glass ware is 40 days -60
My god.
9. a kind of plant leaf blade conservation in vitro method described in claim any one of 1-8 is in Plant seed preserving, plant
Application in genetic test, physiologically active of plant measure and Pathogenicity.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710312053.0A CN107151644A (en) | 2017-05-05 | 2017-05-05 | A kind of plant leaf blade conservation in vitro method and its application |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710312053.0A CN107151644A (en) | 2017-05-05 | 2017-05-05 | A kind of plant leaf blade conservation in vitro method and its application |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102268390A (en) * | 2011-07-07 | 2011-12-07 | 华中农业大学 | Bacillus amyloliquefaciens for generating fungi apoptosis lipopeptide, and preparation method and application thereof |
| CN102726177A (en) * | 2011-04-01 | 2012-10-17 | 华中农业大学 | Method for identifying cotton verticillium by using leaves in vitro |
| CN103392607A (en) * | 2013-08-21 | 2013-11-20 | 江苏省林业科学研究院 | Leaf in-vitro culturing method for miniature ornamental photinia fraseri |
| CN103518625A (en) * | 2013-11-01 | 2014-01-22 | 重庆文理学院 | Tissue culture medium and in-vitro regeneration method for ficus pandurata blade |
| CN105779565A (en) * | 2016-04-14 | 2016-07-20 | 浙江省农业科学院 | Sesquipedalis germplasm rust disease resistance in-vitro identification method and rust fungus liquid used in method |
-
2017
- 2017-05-05 CN CN201710312053.0A patent/CN107151644A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102726177A (en) * | 2011-04-01 | 2012-10-17 | 华中农业大学 | Method for identifying cotton verticillium by using leaves in vitro |
| CN102268390A (en) * | 2011-07-07 | 2011-12-07 | 华中农业大学 | Bacillus amyloliquefaciens for generating fungi apoptosis lipopeptide, and preparation method and application thereof |
| CN103392607A (en) * | 2013-08-21 | 2013-11-20 | 江苏省林业科学研究院 | Leaf in-vitro culturing method for miniature ornamental photinia fraseri |
| CN103518625A (en) * | 2013-11-01 | 2014-01-22 | 重庆文理学院 | Tissue culture medium and in-vitro regeneration method for ficus pandurata blade |
| CN105779565A (en) * | 2016-04-14 | 2016-07-20 | 浙江省农业科学院 | Sesquipedalis germplasm rust disease resistance in-vitro identification method and rust fungus liquid used in method |
Non-Patent Citations (2)
| Title |
|---|
| M SAN JOSÉ等: "Histology of the regeneration of Paulownia tomentosa (Paulowniaceae) by organogenesis", 《REV.BIOL.TROP.》 * |
| 曾翠云: "沙打旺9个品种对沙打旺黄矮根腐病的抗性机理研究及种质特性综合评价", 《兰州大学》 * |
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Application publication date: 20170912 |