CN106577873B - The inducing substance with raising Allelopathy on Oryza sativa L extracted in liquid is planted from barnyard grass - Google Patents
The inducing substance with raising Allelopathy on Oryza sativa L extracted in liquid is planted from barnyard grass Download PDFInfo
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- CN106577873B CN106577873B CN201611083122.7A CN201611083122A CN106577873B CN 106577873 B CN106577873 B CN 106577873B CN 201611083122 A CN201611083122 A CN 201611083122A CN 106577873 B CN106577873 B CN 106577873B
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- barnyard grass
- liquid
- allelopathy
- inducing substance
- oryza sativa
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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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/40—Liliopsida [monocotyledons]
- A01N65/44—Poaceae or Gramineae [Grass family], e.g. bamboo, lemon grass or citronella grass
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- Engineering & Computer Science (AREA)
- Agronomy & Crop Science (AREA)
- Biotechnology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
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Abstract
The invention discloses a kind of inducing substance with raising Allelopathy on Oryza sativa L planted from barnyard grass and extracted in liquid, it is that adsorbing separation is carried out to barnyard grass plantation liquid using macroporous absorbent resin ADS8, there is the inducing substance for improving Allelopathy on Oryza sativa L described in acquisition；The inducing substance is that apolar mixture is remarkably improved the allelopathy of rice wherein containing amides, indoles, phenols, polycyclic class, alcohol compound.
Description
Technical field
Present invention relates particularly to what is extracted in a kind of plantation liquid from barnyard grass to have the inducing substance for improving Allelopathy on Oryza sativa L,
Belong to agroecology field.
Background technology
Inducible system resistance（ISR）It is a kind of phenomenon being widely present in plant kingdom, in addition to utilizing artificial synthesized substance
It induces plant to increase outside anti-adversity ability, other plant can also be induced to enhance degeneration-resistant effect using the substance of plant origin.Barnyard grass
Grass is the Associated weeds of rice, can not only secrete plant hormone, while can also generate the signaling molecule of various plants, induction
Rice enhances allelopathy.Studies have shown that Allelopathy on Oryza sativa L in the presence of barnyard grass enhances, this illustrates can between rice and barnyard grass
There can be chemical communication, so as to perceive the presence of other side mutually.But still for bipartite chemical communication substance at present
It is unclear, also correlative study there is no to report both at home and abroad.
The present invention is detached by planting liquid to barnyard grass, obtains a kind of object that can be induced and improve Allelopathy on Oryza sativa L
Matter, and constituent analysis is carried out to it, mechanism is mutually distinguishable between announcement plant and is acted on by induction raising rice suppression grass, with
Reduction herbicide uses, protect the fields ecological environment, all has important scientific meaning and application potential.
Invention content
The purpose of the present invention is to provide a kind of luring with raising Allelopathy on Oryza sativa L planted from barnyard grass and extracted in liquid
Substance is led, gained inducing substance is apolar mixture, is remarkably improved allelopathic Allelopathy on Oryza sativa L.
To achieve the above object, the present invention adopts the following technical scheme that：
It is a kind of to plant the inducing substance with raising Allelopathy on Oryza sativa L extracted in liquid from barnyard grass, it is to be inhaled using macropore
Attached Resin A DS8 carries out adsorbing separation to barnyard grass plantation liquid, obtains the inducing substance for having and improving Allelopathy on Oryza sativa L；It should
Inducing substance is apolar mixture, wherein containing amides, indoles, phenols, polycyclic class, alcohol compound.
The extracting method of the inducing substance includes the following steps：
1）Barnyard grass plants the collection of liquid：The barnyard grass seed for selecting full grains is sterilized with the liquor natrii hypochloritis of 0.5wt%
It is clean with distilled water flushing after handling 10min, then by seed tiling to sand table, preservative film is covered after adding appropriate distilled water, and
Some apertures are stabbed on preservative film with tweezers, is put into constant incubator and carries out vernalization in 30 DEG C, when barnyard grass was grown to a leaf phase,
The uniform barnyard grass of 120 plants of growing ways of picking（High 4cm or so）, it is transferred into equipped in the 10L suddenly plastic tub of Glan pancebrin
（Plastic tub specification：Long 45cm × wide 35cm × high 15cm）, basin is put in 25 DEG C -35 DEG C outdoor plantations naturally, supplies water weekly
Point and nutrition（To 10L, nutrition completely consumed calculating by 7 days, refilled original nutrient concentrations for direct moisturizing）；Plant 25d extremely
After barnyard grass grows to 5 leaves, nutrient solution in basin is collected, is filtered with 3 layers of filter paper, then through 0.45 μm of membrane filtration, 40 DEG C of rotary evaporations are dense
It is reduced to 120mL（It is the juice equal to one plant of barnyard grass per 1mL）Liquid is planted to get barnyard grass, -20 DEG C are in store for；
2）It is resin activated：Macroporous absorbent resin ADS8 100g are taken, after being activated overnight with soaked in absolute ethyl alcohol, use distilled water
It impregnates, clean to no ethanol flavor, it is spare then to fill column；
3）Barnyard grass plants the adsorbing separation of liquid：10L barnyard grasses plantation liquid upper prop is adsorbed, institute adsorbent 800mL
After the elution of 80wt% ethanol solutions, decompression rotary evaporation is concentrated into 120mL to get the inducing substance, and -20 DEG C are in store for.
The remarkable advantage of the present invention is：To realize the ingredient for being planted to barnyard grass and can induce rice in liquid and improving allelopathy
It is further studied, the present invention carries out adsorbing separation using macroporous absorbent resin ADS8 to barnyard grass plantation liquid, obtains nonpolarity
Inducing substance, obtained material is remarkably improved Allelopathy on Oryza sativa L, can make suppression of the allelopathic rice to weed growth after induction
The allelopathic rice that rate processed does not induce improves 2.35 times.
Specific implementation mode
In order to make content of the present invention easily facilitate understanding, With reference to embodiment to of the present invention
Technical solution is described further, but the present invention is not limited only to this.
Barnyard grass used（Echinochloa crusgalli(L.) Beauv）Seed collection is in University Of Agriculture and Forestry In Fujian field
Experimental field simultaneously preserves 1 year.
Big pore adsorption resin ADS8（Non-polar resin）And ADS21（Polar resin）It is purchased from Nankai University.
1. the acquisition of inducing substance
1.1 barnyard grasses plant the collection of liquid
The barnyard grass seed for selecting full grains uses distilled water after disinfecting 10min with the liquor natrii hypochloritis of 0.5wt%
It rinses well, then will be in seed tiling to sand table（Sand sterilizes 30min prior to 65 DEG C in advance, 3 times repeatedly after cooling）, add appropriate steaming
Preservative film is covered after distilled water, tweezers is used in combination to stab some apertures on preservative film, is put into constant incubator and is carried out vernalization in 30 DEG C,
When barnyard grass was grown to a leaf phase, the uniform barnyard grass of picking growing way（High 4cm or so）, it is transferred into that Glan is sought entirely suddenly equipped with 10L
In the plastic tub of nutrient solution（Plastic tub specification：Long 45cm × wide 35cm × high 15cm）, per 120 plants of basin, basin is put in 25 DEG C -35 DEG C
It is outdoor to plant naturally, moisture and nutrition are supplied weekly（To 10L, nutrition completely consumed calculating by 7 days for direct moisturizing, refilled original
Nutrient concentrations）；After plantation 25d grows to 5 leaves to barnyard grass, nutrient solution in each basin is collected respectively, it is filtered with 3 layers of filter paper
Afterwards, then through 0.45 μm of membrane filtration, 40 DEG C of rotary evaporations are concentrated into 120mL（It is the juice equal to one plant of barnyard grass per 1mL）, obtain
Barnyard grass plants liquid, and -20 DEG C are in store for；
1.2 resin activated
Macroporous absorbent resin each 100g of ADS8 and ADS21 is taken, after being activated overnight respectively with soaked in absolute ethyl alcohol, uses distilled water
It impregnates, clean to no ethanol flavor, it is spare with the populated pillar of both resins respectively.
1.3 barnyard grasses plant the adsorbing separation of liquid
2 parts of barnyard grass plantation liquid, every part of 10L are taken to carry out absorption point after pre-packed ADS8 columns and ADS21 columns respectively
From；Gained efflux decompression rotary evaporation is concentrated into 120mL, and -20 DEG C are in store for；The substance 800mL of ADS8 columns absorption
80wt% ethanol solutions are eluted, and the substance of ADS21 columns absorption is eluted with 30% ethanol solutions of 800mL, two kinds of eluents
Decompression rotary evaporation is concentrated into 120mL respectively, and -20 DEG C are in store for.
2. inducing effect is tested
2.1 isolate inducing paddy rice allelopathies
Outflow concentrate, elution concentrate difference using the outflow concentrate of ADS8 columns, elution concentrate with ADS21 columns
Allelopathic rice PI312777 is induced, every part of dosage is 15mL, is control with barnyard grass plantation liquid induction, experiment is repeated four times
（Mean temperature：34 DEG C of daytime, 25 DEG C at night）.After induction 2 days, rice leaf, -20 DEG C of preservations are collected.To the allelopathic after induction
The blade extract of rice PI312777 carries out biological test, and raw survey measures the root long and plant height of barnyard grass, inhibited with calculating after 4 days
Rate.
2.2 data analysis
Using 11.0 Software on Drawing of SigmaPlot, inhibiting rate, which calculates, presses formula：IR= (1-TR/CKThe % of) × 100 is calculated,
Wherein,IR：Inhibiting rate（Inhibition rate）,TR：Processing group growth indexes（Treatment）,CK：Negative control group is given birth to
Long pointer（Control）.Data are for statistical analysis using DPS2000 softwares, and significant difference analysis is carried out using LSD methods.
3. isolate inducing paddy rice allelopathy test result
Induction experiments the results are shown in Table 1, by table 1 as it can be seen that ADS8 columns elute the rice leaf leaching liquor pair after concentrate induction
The inhibiting rate of barnyard grass root long is 48.95 ± 3.35%, is significantly higher than barnyard grass plantation liquid processing（20.83±5.24%）；ADS21 columns are washed
Rice leaf leaching liquor after de- concentrate induction is 12.80+2.71% to the inhibiting rate of barnyard grass root long, but plants liquid than barnyard grass
It handles slightly lower.It proves after ADS8 post separations, gained elution concentrate has the function of significantly improving allelopathic rice.
Inhibiting rate of the 1 rice leaf leaching liquor of table to barnyard grass root long
Experiment proves, barnyard grass is planted liquid after the separation of non polarity A DS8 macroporous absorbent resins, with gained inducing substance pair
Allelopathic rice is induced, and inhibiting effect of the rice leaf extract to barnyard grass is remarkably improved；And it is inhaled through polarity ADS21 macropores
Attached resin is from rear gained inducing substance to improving Allelopathy on Oryza sativa L almost without effect.
4. isolate GC-MS analyses
4.1 analysis method
According to the Induction experiments of Allelopathy on Oryza sativa L as a result, taking macroporous absorbent resin ADS8（Non-polar resin）Elution it is dense
Contracting liquid 50mL first uses 3 extractions of 300mL ethyl acetate point, each 100mL to merge the ethyl acetate after extraction, 30 DEG C of decompressions
Solution residue 5mL or so is distilled, is poured into 10mL centrifuge tubes, freeze-drying is to being in powder, with 0.5mL ethyl acetate（Chromatography
It is pure）Dissolving, crosses 0.22 μm of filter membrane, and 4 DEG C of placements are spare.
GC-MS conditions：Agilent 7890A/5975C, DB-5 capillary columns（30m×0.25mm）；Carrier gas is helium, stream
Fast 0.8mL/min；Temperature program：50 DEG C of initial column temperature keeps 5min, rises to 180 DEG C with the rate of 10 DEG C/min, keeps
4min, 20 DEG C/min are raised to 220 DEG C, keep 5min, are raised to 280 DEG C with the rate of 5 DEG C/min, keep 5min；Injector temperature
280 DEG C, electronic constant current control, Splitless injecting-Sample, 2 μ L of sampling volume；250 DEG C of interface temperature, 230 DEG C of ion source temperature, level Four
150 DEG C of bar temperature；Full scan：Sweep speed 0.4s/ times, scanning range m/z 15-900.With spectrogram library（NIST98 &
WILEY）Compound is retrieved, is classified to compound, the substance of undetected peak and peak area less than 0.5% is ignored.Chemical combination
The classification of object is classified by the effect of the skeleton of molecular structure.
4.2 testing result
Acetic acid ethyl ester extract to eluting concentrate by ADS8 columns is detected, and obtains 46 compounds（It is shown in Table 2）,
Wherein polycyclic class 6（Peak area 10.64%）, ketone 2（Peak area 2.25%）, nitrile 1（Peak area 1.08%）, alcohols 4
（Peak area 3.06%）, polycyclic aromatic hydrocarbon 2（Peak area 7.20%）, esters 6（Peak area 7.66%）, nitrogen heterocyclic ring 2（Peak
Area 2.51%）, phenols 8（Peak area 11.46%）, oxygen heterocycle 1（Peak area 0.75%）, carboxylic acids 2（Peak area
2.92%）, alkanes 4（Peak area 2.95%）, amides 5（Peak area 43.99%）, indoles 3（Peak area 3.54%）.
Concrete outcome is shown in Table 2.
2 ADS8 columns of table elute the compound in concentrate acetic acid ethyl ester extract
As can be seen from Table 2, amides are the maximum ratios that detected in ADS8 elution concentrate ethyl acetate extraction phases
Substance.Structural analysis shows in 5 amide substances that 2 are unsaturated long-chains, account for the 27.43% of overall material, account for amides
62.35%, remaining 3 be saturation long-chain.
Indoles substance detects 3 altogether, although which accounts for whole only 3.54%, one of substance and
It is detected in barnyard grass plantation liquid identical.
Aldehydes matter totally 8 accounts for the 46.44% of phenols totality, close to one wherein the aldehydes matter containing tertiary butyl accounts for 5.32%
Half amount, this 2 tertiary butyl substances are also detected in barnyard grass plantation.
It can be seen that after non polarity A DS8 macroporous absorbent resin adsorbing separations, the inducing substance of gained is mainly non-pole
The compounds such as property substance, including amides, indoles, phenols.
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 should all belong to the covering scope of the present invention.
Claims (2)
1. a kind of planting the inducing substance with raising Allelopathy on Oryza sativa L extracted in liquid from barnyard grass, which is characterized in that use
Macroporous absorbent resin ADS8 carries out adsorbing separation to barnyard grass plantation liquid, obtains the inducer for having and improving Allelopathy on Oryza sativa L
Matter；Its extracting method specifically comprises the following steps：
1）Barnyard grass plants the collection of liquid：The barnyard grass seed for selecting full grains, is disinfected with the liquor natrii hypochloritis of 0.5wt%
It is clean with distilled water flushing after 10min, then by seed tiling to sand table, add water and urged in 30 DEG C after being capped preservative film
Bud, when barnyard grass was grown to a leaf phase, the uniform barnyard grass of 120 plants of growing ways of picking is transferred into equipped with the 10 L full nutrition of Glan suddenly
In the plastic tub of liquid, in 25 DEG C -35 DEG C outdoor plantations naturally, moisture and nutrition are supplied weekly；Plantation 25d grows to 5 leaves to barnyard grass
Afterwards, the nutrient solution in basin is collected, is filtered with 3 layers of filter paper, then through 0.45 μm of membrane filtration, 40 DEG C of rotary evaporations are concentrated into 120
ML plants liquid to get barnyard grass, and -20 DEG C are in store for；
2）It is resin activated：Take macroporous absorbent resin ADS8 100g, after being activated overnight with soaked in absolute ethyl alcohol, with distilled water immersion,
Then it is spare to fill column to no ethanol flavor for cleaning；
3）Barnyard grass plants the adsorbing separation of liquid：10L barnyard grasses plantation liquid upper prop is adsorbed, institute adsorbent 800mL
After the elution of 80wt% ethanol solutions, decompression rotary evaporation is concentrated into 120mL to get the inducing substance, and -20 DEG C are in store for；
The inducing substance is apolar mixture, wherein containing amides, indoles, phenols, polycyclic class, alcohol compound.
2. a kind of application of inducing substance as described in claim 1 on improving Allelopathy on Oryza sativa L.
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| CN201611083122.7A CN106577873B (en) | 2016-11-30 | 2016-11-30 | The inducing substance with raising Allelopathy on Oryza sativa L extracted in liquid is planted from barnyard grass |
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| CN201611083122.7A CN106577873B (en) | 2016-11-30 | 2016-11-30 | The inducing substance with raising Allelopathy on Oryza sativa L extracted in liquid is planted from barnyard grass |
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| CN109379920B (en) * | 2018-09-21 | 2021-11-02 | 湖南省植物保护研究所 | Method for rapidly breaking barnyard grass seed dormancy and improving germination rate |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102257947A (en) * | 2011-04-29 | 2011-11-30 | 安徽省农业科学院水稻研究所 | Method for screening of paddy rice with weed inhibiting function in laboratories |
| CN103834575A (en) * | 2014-01-15 | 2014-06-04 | 福建农林大学 | Grass-suppressive fungus screened from allelopathic rice soil and application thereof |
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2016
- 2016-11-30 CN CN201611083122.7A patent/CN106577873B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102257947A (en) * | 2011-04-29 | 2011-11-30 | 安徽省农业科学院水稻研究所 | Method for screening of paddy rice with weed inhibiting function in laboratories |
| CN103834575A (en) * | 2014-01-15 | 2014-06-04 | 福建农林大学 | Grass-suppressive fungus screened from allelopathic rice soil and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| 化感水稻苗期不同器官水浸提液及根系分泌物对稗草的化感作用;何海斌等;《中国生态农业学报》;20070330;第15卷(第2期);15 * |
| 杂草对不同水稻品种化感作用的诱导效应;王艳平等;《生态环境》;20050420;第14卷(第2期);250-252 * |
| 稗草浸提液诱导水稻化感抑草作用研究;俞振明等;《现代农业科技》;20130320;109 * |
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