CN106614586A - IDL4 mature polypeptide plant senescence accelerator, preparation method and application - Google Patents
IDL4 mature polypeptide plant senescence accelerator, preparation method and application Download PDFInfo
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- CN106614586A CN106614586A CN201610873606.5A CN201610873606A CN106614586A CN 106614586 A CN106614586 A CN 106614586A CN 201610873606 A CN201610873606 A CN 201610873606A CN 106614586 A CN106614586 A CN 106614586A
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- mature polypeptide
- plant senescence
- morpholines
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- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 131
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 128
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 128
- 101100340320 Arabidopsis thaliana IDL4 gene Proteins 0.000 title claims abstract description 122
- 230000009758 senescence Effects 0.000 title claims abstract description 83
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 2
- 241000196324 Embryophyta Species 0.000 claims description 96
- 239000000243 solution Substances 0.000 claims description 59
- CCIVGXIOQKPBKL-UHFFFAOYSA-N ethanesulfonic acid Chemical compound CCS(O)(=O)=O CCIVGXIOQKPBKL-UHFFFAOYSA-N 0.000 claims description 54
- 239000012530 fluid Substances 0.000 claims description 34
- 235000002637 Nicotiana tabacum Nutrition 0.000 claims description 27
- 238000000034 method Methods 0.000 claims description 24
- 239000010413 mother solution Substances 0.000 claims description 22
- 244000061176 Nicotiana tabacum Species 0.000 claims description 21
- 230000032683 aging Effects 0.000 claims description 20
- 230000008569 process Effects 0.000 claims description 17
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 14
- 239000011521 glass Substances 0.000 claims description 9
- 229920004890 Triton X-100 Polymers 0.000 claims description 8
- 239000013504 Triton X-100 Substances 0.000 claims description 8
- 239000012752 auxiliary agent Substances 0.000 claims description 8
- 239000002904 solvent Substances 0.000 claims description 8
- HNGDOSBFYRVIEY-UHFFFAOYSA-N ethanesulfonic acid;hydrate Chemical compound O.CCS(O)(=O)=O HNGDOSBFYRVIEY-UHFFFAOYSA-N 0.000 claims description 7
- 239000000843 powder Substances 0.000 claims description 7
- 238000003756 stirring Methods 0.000 claims description 7
- 235000009161 Espostoa lanata Nutrition 0.000 claims description 6
- 240000001624 Espostoa lanata Species 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 3
- 239000007921 spray Substances 0.000 claims description 3
- 150000004682 monohydrates Chemical class 0.000 abstract description 6
- 239000001963 growth medium Substances 0.000 abstract description 3
- 238000009630 liquid culture Methods 0.000 abstract description 3
- 230000014634 leaf senescence Effects 0.000 abstract description 2
- 239000004615 ingredient Substances 0.000 abstract 1
- 241000219194 Arabidopsis Species 0.000 description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 241000208125 Nicotiana Species 0.000 description 6
- SMWDFEZZVXVKRB-UHFFFAOYSA-N anhydrous quinoline Natural products N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 description 6
- 229930002875 chlorophyll Natural products 0.000 description 5
- 235000019804 chlorophyll Nutrition 0.000 description 5
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 5
- 238000003860 storage Methods 0.000 description 5
- YNAVUWVOSKDBBP-UHFFFAOYSA-N Morpholine Natural products C1COCCN1 YNAVUWVOSKDBBP-UHFFFAOYSA-N 0.000 description 4
- 150000001413 amino acids Chemical class 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000003306 harvesting Methods 0.000 description 4
- 230000014759 maintenance of location Effects 0.000 description 4
- 101150091470 IDL4 gene Proteins 0.000 description 3
- 238000003483 aging Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 230000007774 longterm Effects 0.000 description 3
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- 108090000623 proteins and genes Proteins 0.000 description 3
- JLIDBLDQVAYHNE-YKALOCIXSA-N (+)-Abscisic acid Chemical compound OC(=O)/C=C(/C)\C=C\[C@@]1(O)C(C)=CC(=O)CC1(C)C JLIDBLDQVAYHNE-YKALOCIXSA-N 0.000 description 2
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 2
- 239000005864 Sulphur Substances 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- FFBHFFJDDLITSX-UHFFFAOYSA-N benzyl N-[2-hydroxy-4-(3-oxomorpholin-4-yl)phenyl]carbamate Chemical compound OC1=C(NC(=O)OCC2=CC=CC=C2)C=CC(=C1)N1CCOCC1=O FFBHFFJDDLITSX-UHFFFAOYSA-N 0.000 description 2
- 230000008827 biological function Effects 0.000 description 2
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- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 238000003208 gene overexpression Methods 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 238000007654 immersion Methods 0.000 description 2
- 230000020429 meristem development Effects 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000005507 spraying Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000012549 training Methods 0.000 description 2
- 229930192334 Auxin Natural products 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 108010028143 Dioxygenases Proteins 0.000 description 1
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 1
- 239000005977 Ethylene Substances 0.000 description 1
- 240000002853 Nelumbo nucifera Species 0.000 description 1
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 1
- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
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- 239000002253 acid Substances 0.000 description 1
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- 239000002671 adjuvant Substances 0.000 description 1
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- 239000007864 aqueous solution Substances 0.000 description 1
- 239000002363 auxin Substances 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
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- 238000003776 cleavage reaction Methods 0.000 description 1
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- 230000001276 controlling effect Effects 0.000 description 1
- 238000012272 crop production Methods 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- FCRACOPGPMPSHN-UHFFFAOYSA-N desoxyabscisic acid Natural products OC(=O)C=C(C)C=CC1C(C)=CC(=O)CC1(C)C FCRACOPGPMPSHN-UHFFFAOYSA-N 0.000 description 1
- QDOXWKRWXJOMAK-UHFFFAOYSA-N dichromium trioxide Chemical compound O=[Cr]O[Cr]=O QDOXWKRWXJOMAK-UHFFFAOYSA-N 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
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- 230000012010 growth Effects 0.000 description 1
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000004643 material aging Methods 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 101150068385 nap gene Proteins 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 125000005478 oxoglutarate group Chemical class 0.000 description 1
- 229930195732 phytohormone Natural products 0.000 description 1
- 230000001323 posttranslational effect Effects 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
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Classifications
-
- 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
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
- A01N37/46—N-acyl derivatives
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biodiversity & Conservation Biology (AREA)
- Ecology (AREA)
- Forests & Forestry (AREA)
- Wood Science & Technology (AREA)
- Agronomy & Crop Science (AREA)
- Botany (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Health & Medical Sciences (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Peptides Or Proteins (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention provides an IDL4 mature polypeptide plant senescence accelerator, a preparation method and application thereof and belongs to the field of plant senescence accelerators. The plant senescence accelerator can accelerate leaf senescence of plants to accelerate senescence of the plants and has no other extra bad phenotypes. A functional ingredient of the plant senescence accelerator is IDL4 mature polypeptide, and an amino acid sequence of the ILD4 mature polypeptide is shown as Pro-Val-Pro-Ala-Ser-Ala-Pro-Ser-Arg-Lys-His-Asn. The IDL4 mature polypeptide plant senescence accelerator comprises the IDL4 mature polypeptide and 2-(N-morpholine) ethanesulfonic solution which is formed by dissolving 2-(N-morpholine) ethanesulfonic monohydrate in 1/2 MS liquid culture medium or MS liquid culture medium. The IDL4 mature polypeptide plant senescence accelerator can be used for accelerating plant senescence.
Description
Technical field
The present invention relates to plant senescence accelerator field, more particularly to a kind of IDL4 mature polypeptides plant senescence accelerator and
Its preparation method and application.
Background technology
Leaf senile is the key factor for affecting the economical characters such as crop yield, quality.By taking Nicotiana tabacum L. as an example, Nicotiana tabacum L. is weight
Want industrial crops, blade is that it harvests organ, as Nicotiana tabacum L. enters late growth stage, blade enters ageing phase, Nicotiana tabacum L. top, in
Portion, lower blade mature period are differed, and generally, are completed using the strategy of different parts different times harvesting in production, are delayed
Collecting time；On the other hand, for example close harvest time of the environmental factorss continuous rainy day be likely to result in Nicotiana tabacum L. " secondary to turn green " shadow
Ring harvesting.Therefore, using aging accelerator, plant leaf blade aging can be promoted, can effectively solving different parts tobacco leaf into
The inconsistent and problem of turning green of ripe degree.
At present, the regulation aspect to phytohormone, auxin, cell point are concentrated mainly on to the application of plant senescence control
Splitting element etc. can suppress leaf senile, ethylene, abscisic acid to promote leaf senile；Artificially slow down Leaf senescence development, on the one hand may be used
To promote the synthesis or metabolism of hormone by reducing aging, on the other hand can also increase the synthesis or metabolism of senescence inhibitory hormones.
The wide and deep development that works of the related biological function explore of aging, the gene quilt of a large amount of promotion leaf seniles
Biological function explore.For example, the dioxygenase gene disappearance that arabidopsiss 2- oxoglutarates are relied on can make leaf senile postpone phenotype；Ground
Sai meter Song (DEX) is induced after the gene overexpression, can render transgenic Arabidopsis leaf aging shift to an earlier date；Arabidopsiss AtNAC families
Member NAP gene overexpressions can make blade do sth. in advance aging, and NAP deletion mutants body then delay aging, equally, in Oryza sativa L.
In the gene can also affect leaf senile；In addition, all controllable leaves such as transcription factor families such as plant tiny RNA, WRKY
The aging of piece.But to current, the SAG reported is really applied directly to being also rarely reported in crop production.
Polypeptide is the small-molecule substance that the class in animal and plant body has regulating and controlling effect, and plant polypeptide take part in multinomial development
Process, such as root meristem development, shoot apical meristem development etc., but whether plant polypeptide participates in aging, has not been reported,
Mature polypeptide is, through post-translational cleavage, modification, to ultimately form the functional molecular of only shorter aminoacid composition by precursor protein,
As part and the Receptor recognition being positioned on cell membrane, enabling signal transduction.Directly outer applying can to plant growing for mature polypeptide
Produce impact, the such as elongation of root.At present applied research of the mature polypeptide on plant especially crop seldom, more will not at present
It applies the research in terms of plant senescence is promoted.
The content of the invention
It is an object of the invention to provide a kind of IDL4 mature polypeptides plant senescence accelerator, preparation method and application.Should
IDL4 mature polypeptide plant senescence accelerator can promote the leaf senile of plant so as to which aging accelerates, and extra without other
Bad phenotype.
One aspect of the present invention provides IDL4 mature polypeptide plant senescence accelerator, the plant senescence accelerator function composition
For IDL4 mature polypeptides, the aminoacid sequence of IDL4 mature polypeptides is Pro-Val-Pro-Ala-Ser-Ala-Pro-Ser-Arg-
Lys-His-Asn。
Preferably, the IDL4 mature polypeptides plant senescence accelerator includes：IDL4 mature polypeptides, 2- (N- morpholines) second sulphur
Acid solution；2- (N- morpholines) the ethyl sulfonic acid solution is dissolved in 1/2MS liquid cultures for 2- (N- morpholines) ethyl sulfonic acid monohydrate
Base or MS fluid mediums.
Preferably, in the IDL4 mature polypeptides plant senescence accelerator, the concentration of IDL4 mature polypeptides is 8-10 μm of ol/
The concentration of L, 2- (N- morpholines) ethyl sulfonic acid is 2.5-3mmol/L, and the pH of 2- (N- morpholines) ethyl sulfonic acid solution is 5.8-6.
It is furthermore preferred that in the IDL4 mature polypeptides plant senescence accelerator, the concentration of IDL4 mature polypeptides is 10 μm of ol/
The concentration of L, 2- (N- morpholines) ethyl sulfonic acid is 3mmol/L, and the pH of 2- (N- morpholines) ethyl sulfonic acid solution is 5.8.
Preferably, IDL4 mature polypeptide plant senescences accelerator of the invention can also add auxiliary agent, it is preferred that auxiliary agent is
Triton X-100, addition is the 1-3 ‰ of plant senescence accelerator total amount.
Another aspect of the present invention provides the preparation method of IDL4 mature polypeptide plant senescence accelerator, specifically, the party
Method comprises the steps：
(1) weigh IDL4 mature polypeptide powder to be dissolved in water or dimethyl sulfoxide solvent, be configured to concentration for 8-
The IDL4 mature polypeptide mother solutions of 10mmol/L are standby；
(2) 1/2MS fluid mediums or MS fluid mediums are prepared, weighs quantitative 2- (N- morpholines) ethyl sulfonic acid one and be hydrated
Thing, in being added to above-mentioned 1/2MS fluid mediums or MS fluid mediums, is configured to the solution of 2.5-3mmol/L, adjusts solution
PH value is 5.8-6, is sufficiently mixed to be uniformly dissolved and obtains 2- (N- morpholines) ethyl sulfonic acid solution；
(3) the IDL4 mature polypeptide mother solutions for obtaining step (1) are added to 2- (N- morpholines) ethyl sulfonic acid obtained in step (2)
In solution, IDL4 mature polypeptides mother solution is 1 with 2- (N- morpholines) ethyl sulfonic acid liquor capacities ratio:1000, Glass rod stirs, and obtains
To IDL4 mature polypeptide plant senescence accelerator.
Preferably, solvent is dimethyl sulfoxide in step (1)；Preferred 1/2MS fluid mediums in step (2)；Step (3)
The Triton X-100 of 1-3 ‰ are added in middle IDL4 mature polypeptides plant senescence accelerator, is stirred with Glass rod.
Preferably, IDL4 mature polypeptide mother solution matching while using in step (1), if not using more than January, -70 DEG C of storages of mother solution
Deposit；If using within January, -20 DEG C of storages of mother solution.
Another aspect of the present invention additionally provides above-mentioned IDL4 mature polypeptides plant senescence accelerator and is promoting plant senescence
The application of aspect, is applied to the blade face of plant, and specifically, application process is：Direct foliage spray is planted with IDL4 mature polypeptides
Thing aging accelerator immersion cotton balls smears blade face, and plant leaf blade full extension period can use, it is preferred that Nicotiana tabacum L. can be in maturation
Early stage foliage applying.
The beneficial effects of the present invention is：(1) IDL4 in IDL4 mature polypeptide plant senescence accelerator of the invention into
Ripe polypeptide synthetic is convenient, obtains facility, can in high volume synthesize；(2) preparation of plant senescence accelerator of the invention and make
Simple with method, field is workable, and for administration personnel need not move through Special Training, operation is convenient, practical；(3) originally
The plant senescence accelerator of invention affects appropriateness to leaf senile, does not result in untoward reaction, beneficial.
Description of the drawings
Fig. 1 is that IDL4 gene delections postpone leaf senile schematic diagram on arabidopsiss；
Fig. 2 processes arabidopsiss and Nicotiana tabacum L. excised leaf result for the IDL4 mature polypeptide plant senescences accelerator of synthetic
Schematic diagram；
Fig. 3 is that IDL4 mature polypeptide plant senescences accelerator processes live body tobacco leaf result schematic diagram.
Specific embodiment
The technical scheme in the embodiment of the present invention will be clearly and completely described below, it is clear that described enforcement
Example is only a part of embodiment of the invention, rather than the embodiment of whole.Based on the embodiment in the present invention, this area is common
The every other embodiment that technical staff is obtained under the premise of creative work is not made, belongs to the model of present invention protection
Enclose.
A kind of IDL4 mature polypeptides plant senescence accelerator, including IDL4 mature polypeptides are embodiments provided,
The aminoacid sequence of IDL4 mature polypeptides is P-V-P-A-S-A-P-S-R-K-H-N, and specifically, aminoacid sequence is Pro-Val-
Pro-Ala-Ser-Ala-Pro-Ser-Arg-Lys-His-Asn, the plant senescence accelerator includes：8-10μmol/L IDL4
Mature polypeptide, 2.5-3mmol/L 2- (N- morpholines) ethyl sulfonic acid solution；2- (N- morpholines) the ethyl sulfonic acid solution is 2- (N-
Quinoline) ethyl sulfonic acid solution monohydrate is dissolved in 1/2MS fluid mediums or MS fluid mediums, 2- (N- morpholines) ethyl sulfonic acid solution
PH be 5.8-6.
It is understood that the concentration of IDL4 mature polypeptides can be 8,9,10 μm of ol/L, 2- (N- morpholines) ethyl sulfonic acid is molten
The concentration of liquid can for 2.5,2.6,2.7,2.8,2.9,3mmol/L, the pH of 2- (N- morpholines) ethyl sulfonic acid solution can for 5.8,
5.9、6。
Specifically, in the above embodiment of the present invention, IDL4 mature polypeptide plant senescences accelerator also includes auxiliary agent, institute
The conven-tional adjuvants that auxiliary agent can be that this area is used for that blade face moistening and blade face to be extended are stated, the auxiliary agent that the present invention is added is Triton
X-100, addition is the 1-3 ‰ of plant senescence accelerator total amount, but it is understood that, any commonly employed blade face in this area
Used additives can be added so as to promote the application of the IDL4 mature polypeptide plant senescence accelerator of the present invention, such as table of this area
Face activating agent, extension agent, wetting agent etc..
Another embodiment of the present invention provides the preparation method of IDL4 mature polypeptide plant senescence accelerator, specifically,
The method comprises the steps：
S1 weighs IDL4 mature polypeptide powder and is dissolved in water or dimethyl sulfoxide solvent, is configured to concentration for 8-
The IDL4 mature polypeptide mother solutions of 10mmol/L are standby.
In this step, because dimethyl sulfoxide is more preferable to the solubility property of polypeptide, the molten of polypeptide can preferably be promoted
Solution, preferably promotes polypeptide to provide corresponding function.(it is difficult in room temperature because IDL4 mature polypeptides have higher requirements to storage temperature
Place more than 24 hours), thus IDL4 mature polypeptide mother solution matching while using, if not using more than January, -70 DEG C of storages of mother solution；
If using within January, -20 DEG C of storages of mother solution.
S2 prepares 1/2MS fluid mediums or MS fluid mediums, weighs quantitative 2- (N- morpholines) ethyl sulfonic acid one and is hydrated
Thing, in being added to above-mentioned 1/2MS fluid mediums or MS fluid mediums, is configured to the solution of 2.5-3mmol/L, adjusts solution
PH value is 5.8-6, is sufficiently mixed to be uniformly dissolved and obtains 2- (N- morpholines) ethyl sulfonic acid solution.
MS or 1/2MS fluid mediums can not only fully dissolve 2- (N- morpholines) ethyl sulfonic acid in this step, so as to ensure
Certain infiltration type is formed in plant surface, the effect for polypeptide provides basis, meanwhile, the fluid medium can also as solvent
There is provided corresponding big moderate-element nutrition, it is ensured that action effect.
The IDL4 mature polypeptide mother solutions that S3 obtains step (1) are added to 2- (N- morpholines) ethyl sulfonic acid obtained in step (2)
In solution, IDL4 mature polypeptides mother solution is 1 with 2- (N- morpholines) ethyl sulfonic acid liquor capacities ratio:1000, Glass rod stirs, and obtains
To IDL4 mature polypeptide plant senescence accelerator.
Can add the Triton X-100 of 1-3 ‰, corresponding auxiliary agent that above-mentioned solution can be promoted to exist as soon as possible in this step
Plant surface is extended and is permeated moistening.This step is IDL4 mature polypeptides mother solution 2- (N- morpholines) ethyl sulfonic acid solution
1000 times of dilution.
Another embodiment of the present invention provides the IDL4 mature polypeptide plant senescence accelerator and is promoting plant senescence side
The application in face, application process is：Direct foliage spray smears blade face, leaves of plants with IDL4 mature polypeptides accelerator immersion cotton balls
Piece full extension period can use, it is preferred that Nicotiana tabacum L. can be in prematuration period foliage applying.
The cotton balls for spraying or soaking the IDL4 mature polypeptide plant senescence accelerator smears plant leaf blade, can make plant growing
On the premise of state is unaffected, plant leaf blade aging, the concordance for being particularly conducive to the asynchronous aging crop of blade is promoted to adopt
Receive, such as Nicotiana tabacum L..
IDL4 mature polypeptides synthetic is convenient in the IDL4 mature polypeptide plant senescence accelerator of above-described embodiment, obtains
It is convenient, can in high volume synthesize；Prepare and using method is simple, field is workable, and personnel need not move through spy for administration
Different training, operation is convenient, practical；The plant senescence inhibitor affects appropriateness to leaf senile, does not result in untoward reaction, especially
Possesses significant application effect on Nicotiana tabacum L..
In order to become apparent from introducing the IDL4 mature polypeptide plant senescence accelerator that the embodiment of the present invention is provided in detail, under
Face will be described in conjunction with specific embodiments.
Embodiment 1
IDL4 mature polypeptide plant senescence accelerator, including：10 μm of ol/L IDL4 mature polypeptides, 3mmol/L 2- (N-
Quinoline) ethyl sulfonic acid solution；2- (N- morpholines) the ethyl sulfonic acid solution is dissolved in 1/ for 2- (N- morpholines) ethyl sulfonic acid solution monohydrate
2MS fluid mediums or MS fluid mediums, the pH of 2- (N- morpholines) ethyl sulfonic acid solution is 5.8.
Preparation method is as follows：
(1) weigh IDL4 mature polypeptide powder to be dissolved in dimethyl sulfoxide solvent, be configured to concentration for 10mmol/L's
IDL4 mature polypeptide mother solutions are standby；
(2) 1/2MS fluid mediums are prepared, weighs quantitative 2- (N- morpholines) ethyl sulfonic acid monohydrate, be added to above-mentioned
In 1/2MS fluid mediums, the solution of 3mmol/L is configured to, it is 5.8 to adjust solution ph, be sufficiently mixed to be uniformly dissolved and obtain
2- (N- morpholines) ethyl sulfonic acid solution；
(3) the IDL4 mature polypeptide mother solutions for obtaining step (1) take 50 μ L and are added to 2- (N- morpholines) obtained in step (2)
In ethyl sulfonic acid solution 50mL, Glass rod stirs, and obtains the IDL4 mature polypeptide plant senescence accelerator of 50mL.
Embodiment 2
IDL4 mature polypeptide plant senescence accelerator, including：8 μm of ol/L IDL4 mature polypeptides, 2.5mmol/L 2- (N-
Morpholine) ethyl sulfonic acid solution；2- (N- morpholines) the ethyl sulfonic acid solution is dissolved in for 2- (N- morpholines) ethyl sulfonic acid solution monohydrate
MS fluid mediums, the pH of 2- (N- morpholines) ethyl sulfonic acid solution is 5.9.
Preparation method is as follows：
(1) weigh IDL4 mature polypeptide powder to be dissolved in dimethyl sulfoxide solvent, be configured to concentration for 8mmol/L's
IDL4 mature polypeptide mother solutions are standby；
(2) MS fluid mediums are prepared, weighs quantitative 2- (N- morpholines) ethyl sulfonic acid monohydrate, be added to above-mentioned MS liquid
In body culture medium, the solution of 2.5mmol/L is configured to, it is 5.9 to adjust solution ph, be sufficiently mixed to be uniformly dissolved and obtain 2- (N-
Morpholine) ethyl sulfonic acid solution；
(3) the IDL4 mature polypeptide mother solutions for obtaining step (1) take 100 μ L and are added to 2- (N- obtained in step (2)
Quinoline) in ethyl sulfonic acid solution 100mL, Glass rod stirs, and obtains the IDL4 mature polypeptide plant senescence accelerator of 100mL.
Embodiment 3
IDL4 mature polypeptide plant senescence accelerator, including：9 μm of ol/L IDL4 mature polypeptides, 2.7mmol/L 2- (N-
Morpholine) ethyl sulfonic acid solution；2- (N- morpholines) the ethyl sulfonic acid solution is dissolved in for 2- (N- morpholines) ethyl sulfonic acid solution monohydrate
MS fluid mediums, the pH of 2- (N- morpholines) ethyl sulfonic acid solution is 6.
Preparation method is as follows：
(1) weigh IDL4 mature polypeptide powder to be dissolved in water, be configured to the IDL4 mature polypeptides that concentration is 9mmol/L female
Liquid is standby；
(2) 1/2MS fluid mediums are prepared, weighs quantitative 2- (N- morpholines) ethyl sulfonic acid monohydrate, be added to above-mentioned
In 1/2MS fluid mediums, the solution of 2.7mmol/L is configured to, it is 6 to adjust solution ph, be sufficiently mixed to be uniformly dissolved and obtain
2- (N- morpholines) ethyl sulfonic acid solution；
(3) the IDL4 mature polypeptide mother solutions for obtaining step (1) take 20 μ L and are added to 2- (N- morpholines) obtained in step (2)
In ethyl sulfonic acid solution 20mL, Glass rod stirs, and adds 1 ‰ Triton X-100, obtains the IDL4 mature polypeptides of 20mL
Plant senescence accelerator.
Embodiment 4
IDL4 mature polypeptide plant senescence accelerator, including：9 μm of ol/L IDL4 mature polypeptides, 2.7mmol/L 2- (N-
Morpholine) ethyl sulfonic acid solution；2- (N- morpholines) the ethyl sulfonic acid solution is dissolved in for 2- (N- morpholines) ethyl sulfonic acid solution monohydrate
MS fluid mediums, the pH of 2- (N- morpholines) ethyl sulfonic acid solution is 6.
Preparation method is as follows：
(1) weigh IDL4 mature polypeptide powder to be dissolved in water, be configured to the IDL4 mature polypeptides that concentration is 9mmol/L female
Liquid is standby；
(2) 1/2MS fluid mediums are prepared, weighs quantitative 2- (N- morpholines) ethyl sulfonic acid monohydrate, be added to above-mentioned
In 1/2MS fluid mediums, the solution of 2.7mmol/L is configured to, it is 6 to adjust solution ph, be sufficiently mixed to be uniformly dissolved and obtain
2- (N- morpholines) ethyl sulfonic acid solution；
(3) the IDL4 mature polypeptide mother solutions for obtaining step (1) take 200 μ L and are added to 2- (N- obtained in step (2)
Quinoline) in ethyl sulfonic acid solution 200mL, Glass rod stirs, and adds 3 ‰ Triton X-100, and the IDL4 for obtaining 200mL is ripe
Polypeptide plant senescence accelerator.
Experimental test
Processed by carrying out IDL4 gene delections on arabidopsiss, referring to Fig. 1.IDL4 bases in pattern dicotyledon arabidopsiss
Because of deletion mutant idl4, with wild type control phenotype analytical under the same conditions：Continuous light grows 25 days, vegetable material life
Comparative observation after long status entirety Phenotypic Observation and picking blade, as a result shows, A, B figure of Fig. 1 is it can be seen that IDL4 genes lack
Lose mutant and postpone leaf senile, meanwhile, identical leaf position (the 6th leaf position) chlorophyll content of different materials is determined using Ethanol Method,
Each material aging characterization parameter is further analyzed, as a result the C such as Fig. 1 schemes, and wild type is low compared with idl4 mutant chlorophyll contents；
In addition, different materials are respectively taken with 30 plants of statistics, leaf senile situation is analyzed, find Fig. 1 the bright wild type of D charts compared with
Idl4 mutant leaf seniles are more obvious, to sum up illustrate, after IDL4 gene delections, plant performance has gone out the symptom for declining in evening,
Further illustrate effect of the IDL4 polypeptides in terms of leaf senile is promoted.
Example 1 prepare IDL4 mature polypeptide plant senescence accelerator, according to application process process respectively arabidopsiss,
The excised leaf of Nicotiana tabacum L. and the Live leaf of Nicotiana tabacum L..
Process 1：Process Arabidopsis leaf：Continuous light grows arabidopsiss 35 days or so, obtains with leaf position excised leaf (the
Six leaf positions), lain on filter paper, polypeptide reagent is uniform, fully sprinkling, meanwhile, compare as 3mM 2- (N- morpholines) second sulphur
Sour (MES) is processed, the process of 1/2MS aqueous solutions, observes phenotype.
Process 2：Nicotiana tabacum L. is prosperous long-term, processes excised leaf, and processing mode is sprinkling, compares and processes 1 same treatment
Process 3：Nicotiana tabacum L. is prosperous long-term, processes Nicotiana tabacum L. Live leaf, and processing mode is to smear tobacco leaf with cotton balls, control with
Process 1 same treatment, Phenotypic Observation.
Referring to Fig. 2, consistent identical leaf position (the 6th lotus throne leaf) Arabidopsis leaf of growing way is chosen with tweezers, relatively regular puts
In culture dish, IDL4 mature polypeptide plant senescence accelerator is processed in the way of spraying, while plus control same way process,
Place under continuous light 5 days, as a result as can be seen that figure A, B show that IDL4 mature polypeptide plant senescences accelerator promotes arabidopsiss
Leaf senile, arabidopsiss excised leaf is sprayed with the blade of IDL4 mature polypeptide plant senescence accelerator after 3 days have obvious senilism
Phenotype, while determining leaf green content using Ethanol Method, finds what control was processed compared with IDL4 mature polypeptide plant senescences accelerator
Chlorophyll content in leaf blades is substantially more；In addition, figure C shows that IDL4 mature polypeptide agings accelerator promotes tobacco leaf aging, choose
The consistent blade of prosperous long-term Nicotiana tabacum L. growing way, using IDL4 agings accelerator and control sprinkling, Nicotiana tabacum L. excised leaf has visible after 5 days
Senilism character.To sum up show, IDL4 mature polypeptide plant senescence accelerator plays the role of to promote aging to Vitro Plant blade.
Referring to Fig. 3, to be grown in continuous light, the Ben's Nicotiana tabacum L. that about 25 days are grown under the conditions of 27 DEG C is material, and cotton balls soaks
Bubble is respectively 5 minutes in IDL4 mature polypeptides plant senescence accelerator working solution and control so as to be impregnated with completely, the cotton for processing
Ball processes live body tobacco leaf visible senilism character after 13 days, referring to figure A；Meanwhile, using Ethanol Method determine smear IDL4 into
13,21 days tobacco leaf chlorophyll contents after ripe polypeptide plant senescence accelerator and control, as a result show compared with the control, IDL4
Tobacco leaf chlorophyll content after mature polypeptide plant senescence accelerator is processed all is reduced, referring to figure B.To sum up show, IDL4
Mature polypeptide plant senescence accelerator is obviously promoted aging to live plant blade really.
Claims (10)
1.IDL4 mature polypeptide plant senescence accelerator, it is characterised in that the function of IDL4 mature polypeptide plant senescence accelerator
Composition is IDL4 mature polypeptides, and the aminoacid sequence of IDL4 mature polypeptides is Pro-Val-Pro-Ala-Ser-Ala-Pro-Ser-
Arg-Lys-His-Asn。
2. IDL4 mature polypeptides plant senescence accelerator according to claim 1, it is characterised in that the IDL4 is ripe more
Peptide plant senescence accelerator includes：IDL4 mature polypeptides, 2- (N- morpholines) ethyl sulfonic acid solution；2- (N- morpholines) ethyl sulfonic acid is molten
Liquid is dissolved in 1/2MS fluid mediums or MS fluid mediums for 2- (N- morpholines) ethyl sulfonic acid monohydrate.
3. IDL4 mature polypeptides plant senescence accelerator according to claim 2, it is characterised in that the IDL4 is ripe more
In peptide plant senescence accelerator, the concentration of IDL4 mature polypeptides is 8-10
μm ol/L, the concentration of 2- (N- morpholines) ethyl sulfonic acid is 2.5-3mmol/L, and the pH of 2- (N- morpholines) ethyl sulfonic acid solution is 5.8-
6。
4. IDL4 mature polypeptides plant senescence accelerator according to claim 3, it is characterised in that the IDL4 is ripe more
In peptide plant senescence accelerator, the concentration of IDL4 mature polypeptides is 10 μm of ol/L, and the concentration of 2- (N- morpholines) ethyl sulfonic acid is
The pH of 3mmol/L, 2- (N- morpholines) ethyl sulfonic acid solution is 5.8.
5. the IDL4 mature polypeptide plant senescence accelerator according to claim 1-4 any one, it is characterised in that this
Bright IDL4 mature polypeptide plant senescence accelerator also contains auxiliary agent.
6. IDL4 mature polypeptides plant senescence accelerator according to claim 5, it is characterised in that the auxiliary agent is
Triton X-100, addition is the 1-3 ‰ of aging accelerator total amount.
7. the preparation method of the IDL4 mature polypeptide plant senescence accelerator described in claim 1-6 any one, its feature exists
In comprising the steps：
(1) weigh IDL4 mature polypeptide powder to be dissolved in water or dimethyl sulfoxide solvent, be configured to concentration for 8-10mmol/L
IDL4 mature polypeptide mother solutions it is standby；
(2) 1/2MS fluid mediums or MS fluid mediums are prepared, weigh quantitative 2- (N- morpholines) ethyl sulfonic acid monohydrate,
In being added to above-mentioned 1/2MS fluid mediums or MS fluid mediums, the solution of 2.5-3mmol/L is configured to, adjusts pH value of solution
It is worth for 5.8-6, is sufficiently mixed to be uniformly dissolved and obtains 2- (N- morpholines) ethyl sulfonic acid solution；
(3) the IDL4 mature polypeptide mother solutions for obtaining step (1) are added to 2- (N- morpholines) ethyl sulfonic acid solution obtained in step (2)
In, IDL4 mature polypeptides mother solution is 1 with 2- (N- morpholines) ethyl sulfonic acid liquor capacities ratio:1000, Glass rod stirs, and obtains
IDL4 mature polypeptide plant senescence accelerator.
8. the preparation method of IDL4 mature polypeptides plant senescence accelerator according to claim 7, it is characterised in that step
(1) solvent is dimethyl sulfoxide in；Fluid medium is 1/2MS fluid mediums in step (2)；IDL4 is ripe in step (3)
The Triton X-100 of 1-3 ‰ are added in polypeptide plant senescence accelerator, is stirred with Glass rod.
9. the application of the IDL4 mature polypeptide plant senescence accelerator described in claim 1-6 any one, it is characterised in that apply
For the blade face of plant, application process is：Direct foliage spray soaks cotton balls with IDL4 mature polypeptide plant senescences accelerator
Blade face is smeared, plant leaf blade full extension period can use.
10. the application of IDL4 mature polypeptides plant senescence accelerator according to claim 9, it is characterised in that the plant
Thing is Nicotiana tabacum L., in Nicotiana tabacum L. prematuration period foliage applying.
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| CN112400880A (en) * | 2020-11-27 | 2021-02-26 | 中国农业科学院烟草研究所 | IDL6 mature polypeptide plant aging promoter, and preparation method and application thereof |
| CN112400881A (en) * | 2020-11-27 | 2021-02-26 | 中国农业科学院烟草研究所 | IDL7 mature polypeptide plant aging promoter, and preparation method and application thereof |
| WO2021047467A1 (en) * | 2019-09-11 | 2021-03-18 | 中国农业科学院烟草研究所 | Application of quercetin in plant senescence promoter |
| CN112586500A (en) * | 2020-11-27 | 2021-04-02 | 中国农业科学院烟草研究所 | AtPep1 mature polypeptide plant senescence promoter, preparation method and application thereof |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2021047467A1 (en) * | 2019-09-11 | 2021-03-18 | 中国农业科学院烟草研究所 | Application of quercetin in plant senescence promoter |
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| CN112400880A (en) * | 2020-11-27 | 2021-02-26 | 中国农业科学院烟草研究所 | IDL6 mature polypeptide plant aging promoter, and preparation method and application thereof |
| CN112400881A (en) * | 2020-11-27 | 2021-02-26 | 中国农业科学院烟草研究所 | IDL7 mature polypeptide plant aging promoter, and preparation method and application thereof |
| CN112586500A (en) * | 2020-11-27 | 2021-04-02 | 中国农业科学院烟草研究所 | AtPep1 mature polypeptide plant senescence promoter, preparation method and application thereof |
| WO2022095963A1 (en) * | 2020-11-27 | 2022-05-12 | 中国农业科学院烟草研究所 | Idl6 mature polypeptide plant aging promoter, preparation method therefor and use thereof |
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