CN105981720A - Polar auxin transport accelerant and application thereof - Google Patents
Polar auxin transport accelerant and application thereof Download PDFInfo
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- CN105981720A CN105981720A CN201610074225.0A CN201610074225A CN105981720A CN 105981720 A CN105981720 A CN 105981720A CN 201610074225 A CN201610074225 A CN 201610074225A CN 105981720 A CN105981720 A CN 105981720A
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- scutellarein
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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
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
- A01N43/16—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
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Abstract
The invention discloses a polar auxin transport accelerant which comprises scutellarin or scutellarein which have functions of accelerating polar auxin transport (PAT) and resisting the inhibition function of a PAT inhibitor TIBA to root elongation, and discloses application of scutellarin and scutellarein in preparing the PAT inhibitor, and scutellarein in preparing a root elongation accelerant. The phenomenon that scutellarin and scutellarein are secondary metabolites with an acceleration function on PAT and are plant sourced PAT accelerants are discovered for the first time, scutellarin and scutellarein are used in the field of plants for the first time, a new prospect that effects of secondary metabolites are brought into play in plant physiology is widened, and relatively greatervalues of scutellarin and scutellarein can be made.
Description
Technical field:
The invention belongs to auxin field, in particular it relates to two kinds promote auximone polar translocation and delay
Solve inhibitor to the flavonoid baicalin of auximone polar translocation inhibitory action and scutellarein and raw preparation plant
Application in long element polar translocation accelerator.
Background technology:
In plant roots growth course, hormone regulating and controlling has played very important effect (Petricka et al.2012), especially
Its Polar Transport of Auxin (polar auxin transport, PAT) is the most crucial to plant roots morphogenesis.Auxin is
Regulating growth of plants and the important hormone of response external environment, including taking root, bloom, tiller, reply low temperature, tackle saline and alkaline
Deng, PAT is then that auxin responds endogenous and exogenous signals and forms the important channel (Vanneste that polar contribution is relied on
And Friml 2009), study confirmation PAT (Ubeda-Tomas et most important to plant roots morphogenesis
al.2012).The effect of plant self has been attracted more and more by Recent study and appliable plant secondary metabolite
Researcher is paid close attention to, if because can use the growth regulator of plant source in agricultural production, will be more easy to be connect by ecosystem
Receive, the most degradable do not result in residual after using, agricultural product security problem can be improved to a certain extent.Most important of which just finds
It is Herba Lobeliae Chinensis lactone as the impact (Gomez-Roldan et al.2008) on plant branching of a kind of novel plant hormone, enters
And opened up it in the developmental application of gramineous crop plant type such as Oryza sativa L., Semen Tritici aestivi.And Herba Lobeliae Chinensis lactone is exactly based on regulation and control PAT
Play (the Domagalska and Leyser 2011) of its effect, thus screening PAT adjusting control agent will have in agricultural production
Wide application prospect.
At present, prior art there are no the aglycon scutellarein report as PAT adjusting control agent of baicalin and baicalin
Road, does not has it to have the antagonism PAT inhibitor TIBA report to the effect of PAT depression effect yet.
Summary of the invention:
It is an object of the invention to provide two kinds of flavonoid baicalins with auximone polar translocation facilitation
And scutellarein, and provide baicalin or scutellarein application in preparing PAT accelerator.Another object of the present invention
It is to provide two kinds of flavonoid baicalins and scutellarein, there is the antagonism PAT inhibitor TIBA effect to PAT depression effect.
In order to realize the above-mentioned purpose of the present invention, the invention provides following technical scheme:
The accelerator of auximone polar translocation, it contains baicalin or scutellarein.
According to described accelerator, it is with baicalin or scutellarein as sole active agent.
According to described accelerator, in wherein said accelerator, the concentration of baicalin or scutellarein is 100 μMs.
Polar Transport of Auxin albumen PIN1 is promoted according to described accelerator, wherein said baicalin or scutellarein
Express with PIN2.
Promote that auxin response promoter DR5 exists according to described accelerator, wherein said baicalin or scutellarein
The tip of a root is expressed, and strengthens auxin and accumulates at root tip.
According to described accelerator, wherein said baicalin or scutellarein antagonism PAT inhibitor Triiodobenzoic acid pair
The suppression of root elongation.
Baicalin and scutellarein application in preparing Polar Transport of Auxin accelerator, described accelerator is with Radix Scutellariae
Element or scutellarein are active component.
Baicalin and scutellarein application in preparing Polar Transport of Auxin accelerator, described accelerator is with Radix Scutellariae
Element or scutellarein are sole active agent, and concentration is respectively 100 μMs.
Scutellarein application in preparing root elongation accelerator, wherein scutellarein has root elongation to promote effect.
Scutellarein application in preparing root elongation accelerator, wherein the concentration of wild Radix Scutellariae promotes that root is stretched when being 50 μMs
Long.
Compared with prior art, the present invention possesses following excellent benefit:
Baicalin has another name called scutellarin, is the main active in Erigeron breviscapus (Vant.) Hand.-Mazz., and scutellarein is its aglycon.In this Shen
During the patent of invention that early stage of asking someone has authorized " promotes parahormone and the application (ZL201210012269.2) thereof of plant roots elongation "
Illustrate baicalin have significant root elongation promote effect, and can antagonism 2,4-D and two kinds of hormone high concentrations of (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate
Time to root elongation inhibitory action.Baicalin can remarkably promote root and extend, and the suppression that root is extended by energy other hormones of antagonism,
It has also been found that the aglycon scutellarein of baicalin also has the root elongation similar with baicalin promotes effect.
It turned out multiple natural flavonoid material such as naringenin, kaempferol etc. about the existing research of PAT can regulate and control
PAT (Jacobs and Rubery 1988), so very likely baicalin and scutellarein are also to play it by regulation and control PAT
Root elongation promotes effect.Based on above-mentioned basis, in addition screening PAT adjusting control agent no matter to the basic research of growth and development of plants or
The application of agricultural production control accurate is the most significant, so exploring baicalin and the scutellarein regulating and controlling effect to PAT
It is the work being rich in novelty and development prospect.
In the present invention, two kinds of Secondary metabolites baicalins and new effect of scutellarein, i.e. promote PAT, be first
Find has the secondary metabolite to PAT with facilitation, thus is secondary metabolite regulation and control PAT research and application
Important breakthrough in field.And baicalin and the scutellarein energy antagonistic inhibitors TIBA inhibitory action to PAT, it is right to illustrate
The regulation and control of PAT can be used in combination the suitableeest regulation and control realizing heterogeneous expectations effect by baicalin or scutellarein with TIBA.This
The bright announcement that baicalin and scutellarein promote PAT effect, provides the PAT accelerator of plant source first, is not only regulation and control
Growth and development of plants provides new selection, and a piece of brand-new prospect has been opened up in the application for baicalin and scutellarein,
Promote Secondary metabolites in terms of plant physiology, play biological effect, be significant.
Accompanying drawing illustrates:
100 μMs of baicalins of Fig. 1 a or scutellarein promote Polar Transport of Auxin albumen PIN1 to express, and (PIN1 coupling is reported
Gene GFP, so green fluorescent protein strengthens instruction PIN1 and expresses enhancing)；
100 μMs of baicalins of Fig. 1 b or scutellarein promote Polar Transport of Auxin albumen PIN2 to express, and (PIN2 coupling is reported
Gene GFP, so green fluorescent protein strengthens instruction PIN2 and expresses enhancing)；
100 μMs of baicalins of Fig. 2 or scutellarein strengthen auxin response promoter DR5 and respond (DR5 promoter at the tip of a root
Coupling Reporter gene GUS, so GUS dye blueness strengthens instruction DR5 response and strengthens, i.e. auxin accumulation increases)；
(TIBA suppression root is stretched in the suppression that root is extended by 100 μMs of baicalins of Fig. 3 or scutellarein antagonism PAT inhibitor TIBA
Long, but the effect being separately added into baicalin or scutellarein TIBA suppression root elongation in culture medium significantly attenuates)；
50 μMs of scutellarein of Fig. 4 promote the elongation of arabidopsis root and promote that effect is similar with 50 μMs of baicalins.
Detailed description of the invention:
Below in conjunction with the accompanying drawings, further illustrate the essentiality content of the present invention with embodiments of the invention, but not
The present invention is limited with this.
Method in following embodiment, if no special instructions, is conventional method.Test material used in following embodiment
Material, if no special instructions, is and is commercially available from routine biochemistry molecular agents company.
Embodiment 1:
The preparation of baicalin:
Title: baicalin/scutellarin/4-hydroxyl Baicalin
(scutellarin/Scutellarein 4'-methyl ether 7-glucuronide)
Molecular formula: C21H18O12
Structural formula:
Preparation: short booth Herba Erigerontis aceris (Erigeron breviscapus (vant) Hand Mass) pulverizing medicinal materials → 70% ethanol
Percolation → concentrated extracting solution → concentrated solution is crossed macroporous resin → 50% ethanol elution → concentrate eluant → concentrated solution hydrochloric acid and is adjusted
PH value to 1-2 → standing 24 hours crystallization → filter crystalline substance → methanol eddy recrystallization (repeatedly) → scutellarin sterling.(1kg is short
Booth Herba Erigerontis aceris medical material prepares scutellarin sterling 3g)
The preparation of scutellarein:
Title: scutellarein/4', 5,6,7-kaempferol
(scutellarein/4',5,6,7-Tetrahydroxyflavone)
Molecular formula: C15H10O6
Structural formula:
Preparation: weigh baicalin 50mg in round-bottomed flask → add 100mL containing 80% ethanol (v/v), 20% concentrated sulphuric acid
(m/v) solution → nitrogen protection lower heated and stirred 3.5 hours → reactant liquor of reaction is poured into 200mL deionized water, separates out light yellow
Precipitation → standing leaching precipitation after 1 hour, deionized water is washed till neutrality → vacuum drying 24 hours, obtains scutellarein crude product
28.1mg → crude product is dissolved in a small amount of ethanol, mixes with 50mg silica gel, flings to solvent, and upper silicagel column → petrol ether/ethyl acetate/
Acetic acid solvent system (5/3/0.4, volume ratio) quickly drip washing → merging same composition, evaporated under reduced pressure → ethanol one water recrystallization →
Obtain scutellarein sterling 27.8mg.
Embodiment 2:
Baicalin and the expression of scutellarein promotion Polar Transport of Auxin albumen PIN1 and PIN2:
PIN1 and PIN2 is responsible for two Major Members of protein family of Polar Transport of Auxin, PIN1:GFP and PIN2:
GFP is that plant Arabidopsis thaliana wild type (Arabidopsis thalianaecotype Columbia, Col) is that background is divided in mode
Other transgenic PIN1 merges reporter gene green fluorescent protein GFP and PIN2 and merges the mutant of reporter gene GFP.With PIN1:
GFP and PIN2:GFP mutant is experiment material, and the method that application filter paper is cultivated is cultivated and processed.By baicalin and scutellarein
Powder is dissolved separately in DMSO to final concentration of 0.1mol/L, then obtains 100 μMs of baicalins with deionized water dilution and open country is yellow
A kind of reed mentioned in ancient books element working solution, contrast solution then dilutes DMSO by same ratio with demineralised water.By 100 μMs of baicalins and scutellarein and
Contrast solution is separately added into 9cm culture dish, every ware 5mL, culture dish middle berth three metafiltration paper, is uniformly sowed on filter paper by seed,
Each process 3 repetition.In 4 DEG C of refrigerator dark vernalization 2 days, it is then transferred to culturing room and cultivates.Condition of culture is photoperiod 14h
Day/10h night, day temperature 22 DEG C, night temperature 19 DEG C, light intensity 120 μm ol m-2 s-1, humidity 50%-60%.Cultivate one week
After, from the culture dish of different disposal, take seedling at random under laser confocal scanning microscope, observe PIN1 and PIN2 albumen exist
The expression of root.Experimental result shows, compare comparison 100 μMs of baicalins and scutellarein can be obviously enhanced PIN1:GFP and
The intensity of PIN2:GFP mutant root tip green fluorescence, illustrates that 100 μMs of baicalins and scutellarein promote auxin polarity fortune
Egg output white PIN1 and PIN2 protein expression (Fig. 1 a, b).
Embodiment 3:
Baicalin and scutellarein strengthen auxin and accumulate at root tip:
DR5 is an engineered promoter comprising auxin response element, by driving in the presence of auxin
The expression of downstream reporter gene can react auxin distribution in plant and accumulation, and DR5::GUS is wild with arabidopsis
Type Col is the mutant that background transgenic DR5 merges Reporter gene GUS.With DR5::GUS mutant as material, by 100 μMs of Huangs
A kind of reed mentioned in ancient books element and scutellarein and contrast solution are separately added into 9cm culture dish, every ware 5mL, and culture dish middle berth three metafiltration paper, by seed
Uniformly sowing is on filter paper, each process 3 repetition.In 4 DEG C of refrigerator dark vernalization 2 days, it is then transferred to culturing room and cultivates.Training
Foster condition is photoperiod 14h day/10h night, day temperature 22 DEG C, night temperature 19 DEG C, light intensity 120 μm ol m-2 s-1, humidity
50%-60%.Preparation GUS dyeing liquor: take 40mL 100mM phosphate buffer (PBS, pH7.0) and be sequentially added into 0.01861g
EDTA, 0.01211195g six cyano ferrum (II) acid potassium, 0.016462g six cyano ferrum (III) acid potassium, be settled to 50mL with PBS,
Add 1%Triton-X100, keep in Dark Place.After one week, from the culture dish of different disposal, take seedling at random, be immersed in 1mL and contain
In the GUS dyeing liquor of 1mg/mL X-gluc, 37 DEG C of lucifuges dyeing 3-5h are until root visible blue.Outwell dyeing liquor, add
95% ethanol rinse 3 times makes background decolour.In basis of microscopic observation tip of a root staining conditions, and take pictures.Experimental result shows, compares
Compare 100 μMs of baicalins and scutellarein significantly increases DR5::GUS mutant root tip GUS and contaminates blue region, illustrate 100 μMs
Baicalin and scutellarein strengthen auxin and accumulate (Fig. 2) at root tip.
Embodiment 4:
The suppression that root is extended by baicalin and scutellarein antagonism PAT inhibitor TIBA:
With arabidopsis wild type Col as material, add 1% agar as culture medium with nitrogen stress MS, in aseptic bar after Seed sterilization
Uniformly it is seeded in culture medium under part, in 4 DEG C of refrigerator dark vernalization 2 days, is then transferred to culturing room and vertically cultivates.Condition of culture
For photoperiod 14h day/10h night, day temperature 22 DEG C, night temperature 19 DEG C, light intensity 120 μm ol m-2 s-1, humidity 50%-
60%.Add culture medium based on 1% agar by nitrogen stress MS, add 1%DMSO (comparison), 20 μMs of TIBA, 20 μMs of TIBA+ respectively
100 μMs of baicalins, 20 μMs of TIBA+100 μM of scutellarein prepare transport medium.After 5 days, seedling is shifted under aseptic condition
It is as the criterion proper alignment seedling labelling tip of a root position with the tip of a root in various process culture medium, after continuing vertically to cultivate 9 days, observes
Root extends, and takes pictures.Experimental result shows, 20 μMs of TIBA seriously inhibit root to extend, and are simultaneously introduced 20 μMs of TIBA and 100
After μM baicalin or scutellarein, although compare comparison root elongation and nevertheless suffer from slightly suppressing, but the elongation of its root is significantly faster than list
Solely add the process of 20 μMs of TIBA, the suppression (figure that root is extended by baicalin and scutellarein antagonism PAT inhibitor TIBA is described
3)。
Embodiment 5:
Scutellarein promotes that root extends:
With arabidopsis wild type Col as material, the method that application filter paper is cultivated is cultivated and is processed.By baicalin and wild Radix Scutellariae
Element powder is dissolved separately in DMSO to final concentration of 0.1mol/L, then obtains 50 μMs of baicalins with deionized water dilution and open country is yellow
A kind of reed mentioned in ancient books element working solution, contrast solution then dilutes DMSO by same ratio with demineralised water.By 50 μMs of baicalins and scutellarein and
Contrast solution is separately added into 9cm culture dish, every ware 5mL, culture dish middle berth three metafiltration paper, is uniformly sowed on filter paper by seed,
Each process 3 repetition.In 4 DEG C of refrigerator dark vernalization 2 days, it is then transferred to culturing room and cultivates.Condition of culture is photoperiod 14h
Day/10h night, day temperature 22 DEG C, night temperature 19 DEG C, light intensity 120 μm ol m-2 s-1, humidity 50%-60%.After 7 days, often
Individual process takes seedling at random and observes root length, and respectively takes 10 Seedlings and unfold root, and marshalling is taken pictures.Experimental result shows, 50 μMs of open countries
Baicalin processes arabidopsis root and rises appreciably, and the degree that root increases processes similar to 50 μMs of baicalins, and scutellarein is described
The same with baicalin have root elongation promotion effect (Fig. 4).
Claims (10)
1. the accelerator of auximone polar translocation, it is characterised in that it contains baicalin or scutellarein.
Accelerator the most according to claim 1, it is characterised in that it is with baicalin or scutellarein as sole active agent.
Accelerator the most according to claim 1 and 2, it is characterised in that baicalin or scutellarein in described accelerator
Concentration is 100 μMs.
4. according to the accelerator described in claim 1 or 2 or 3, it is characterised in that described baicalin or scutellarein promote raw
Long element polar translocation albumen PIN1 and PIN2 expresses.
5. according to the accelerator described in claim 1 or 2 or 3, it is characterised in that described baicalin or scutellarein promote raw
Long element response promoter DR5 is expressed at the tip of a root, strengthens auxin and accumulates at root tip.
6. according to the accelerator described in claim 1 or 2 or 3, it is characterised in that described baicalin or scutellarein antagonism PAT
The suppression that root is extended by inhibitor Triiodobenzoic acid.
7. baicalin and scutellarein application in preparing Polar Transport of Auxin accelerator, it is characterised in that described promotion
Agent is with baicalin or scutellarein as active component.
8. baicalin and scutellarein application in preparing Polar Transport of Auxin accelerator, it is characterised in that described promotion
Agent is with baicalin or scutellarein as sole active agent, and concentration is respectively 100 μMs.
9. scutellarein application in preparing root elongation accelerator.
10. scutellarein application in preparing root elongation accelerator, it is characterised in that the concentration of scutellarein is 50 μMs.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107365736A (en) * | 2017-08-07 | 2017-11-21 | 中国科学院昆明植物研究所 | A kind of inhibitor of plant root tip stem cell Asymmetric division regulatory pathway and its application |
| CN107460155A (en) * | 2017-08-07 | 2017-12-12 | 中国科学院昆明植物研究所 | Two kinds of accelerator of plant root tip stem cell Asymmetric division regulatory pathway and its application |
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| JPH0418019A (en) * | 1990-05-11 | 1992-01-22 | Tsumura & Co | Sialidase inhibitor |
| CN102613185A (en) * | 2012-01-16 | 2012-08-01 | 中国科学院昆明植物研究所 | Parahormone promoting plant root elongation and application thereof |
-
2016
- 2016-02-03 CN CN201610074225.0A patent/CN105981720A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0418019A (en) * | 1990-05-11 | 1992-01-22 | Tsumura & Co | Sialidase inhibitor |
| CN102613185A (en) * | 2012-01-16 | 2012-08-01 | 中国科学院昆明植物研究所 | Parahormone promoting plant root elongation and application thereof |
Non-Patent Citations (3)
| Title |
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| 张旭东等: "灯盏乙素促进根伸长的类激素效应及其机制研究", 《生态文明建设中的植物学：现在与未来——中国植物学会第十五届会员代表大会暨八十周年学术年会论文集——第3分会场：植物分子生物学与基因组学》 * |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107365736A (en) * | 2017-08-07 | 2017-11-21 | 中国科学院昆明植物研究所 | A kind of inhibitor of plant root tip stem cell Asymmetric division regulatory pathway and its application |
| CN107460155A (en) * | 2017-08-07 | 2017-12-12 | 中国科学院昆明植物研究所 | Two kinds of accelerator of plant root tip stem cell Asymmetric division regulatory pathway and its application |
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