CN106941810A - A kind of method of the lower Germination of Soybean Seed of raising NaCl stress - Google Patents
A kind of method of the lower Germination of Soybean Seed of raising NaCl stress Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
- A01C1/08—Immunising seed
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- 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
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Abstract
The invention discloses a kind of method of the lower Germination of Soybean Seed of raising NaCl stress, comprise the following steps：Choose natural air drying after, size is homogeneous, full grains, the soya seeds of no disease and pests harm, it is standby；Soya seeds surface sterilization is carried out with pasteurization liquid, then medicining liquid dipping is cleaned 4 times with the distilled water of sterilizing；In under constant temperature non-illuminated conditions, with the 24h of fluorine pyridine ketone solution processed soybeans seed 12；Soya seeds after processing are planted in soil, cultivation temperature is 20 25 DEG C, during germination, appropriate running water is sprayed at regular time and quantity, seed is sprouted through 2~10d.0.05 0.15 μm of ol/L of present invention fluorine pyridine ketone solution can significantly improve the germination rate of the lower soya seeds of NaCl stress；Agricultural production efficiency on salt-soda soil can be improved.
Description
Technical field
The invention belongs to agriculture applied technology field, specifically, it is related to a kind of lower soya seeds of NaCl stress that improve and sprouts
The method of hair.
Background technology
The soil salinization and secondary salinization have had a strong impact on agricultural production and ecological environment.According to the World Food Programme not
Statistics completely, by 2015, the whole world had more than 20% soil by different degrees of salination.For plant,
Salt stress phenomenon can occur for saliferous soil growth, such as seed germination rate and emergence rate are seriously reduced.Under salt stress, plant
Can external morphology and it is interior a series of change occurs in physio-biochemical characteristics, as sprout be suppressed, plant blade area
Reduce, the number of blade reduces, ABA content rises, and is unfavorable for the normal growth of plant.So, crop seed how is improved in salt
The lower germination rate of stress, agricultural production is carried out on wetland with saline-alkaline, be present and important subject that the future of agriculture is produced.
Soybean (Glycine max L.) is a kind of legume for originating in East Asia, now wide as main oil crops
It is general to be planted in including the U.S., Brazil, Argentina, India and China etc. all over the world, it is plant main in current world wide
Protein and oil plant source.Contain a variety of amino acid needed by human in soybean, at the same also containing abundant protein, fat,
Mineral nutrient element, can be widely used for the fields such as food, health care.Meanwhile, soybean is also important feed resource, its quality quality
It is directly connected to the quality of the food such as meat, egg, milk.Therefore, increase soybean yields, lifting soybean kernel quality are occupied to improving China
People's living standard, increases farmers' income, and ensures national food security, promotes agricultural and ecological sustainable development, all with non-
Often important meaning.China is the area of origin of soybean, but due to being collected in germ plasm resource, the field such as breed breeding, transgenosis
Research is started late, and is made slow progress, and causes Chinese soybean gradually to fall behind in terms of the control of yield, quality and production cost,
Have to by a large amount of imports meet domestic Soybean production needs.Therefore, develop and revitalize China's Soybean Industry, for lifting
Living standards of the people, safeguard that national food security is significant.
In agricultural production, the sprouting of seed and emergence rate are the key factors for determining crop yield height.If planted
Sub- germination rate is low, and Seedling emergence rate can be caused out low, and the reduction of effective strain number can cause crop yield to reduce.But soya seeds
Containing higher grease and protein, it is unfavorable for seed sprouting under adverse environmental factor, these situations can all cause soya seeds to be sprouted
Hair rate is reduced, so as to influence effective strain number and yield.In addition, food industry soybean processing product to the nutritional ingredient of soybean and
Sprouting situation has very high requirement.Therefore, molecule and physiological mechanism during soybean germination is worth further investigation.
The content of the invention
In view of this, there is provided a kind of lower Germination of Soybean Seed of raising NaCl stress for the problem of present invention is directed to above-mentioned
Method, in agricultural production, can apply appropriate ABA synthetic inhibitors-fluorine pyridine ketone to improve the bud ratio of soybean.
In order to solve the above-mentioned technical problem, the invention discloses a kind of side of the lower Germination of Soybean Seed of raising NaCl stress
Method, comprises the following steps：
1) Seeds preprocess：Choose selection size after natural air drying homogeneous, full grains, the soya seeds of no disease and pests harm,
It is standby；
2) soya seeds surface sterilization is carried out with pasteurization liquid, then medicining liquid dipping cleans 4 with the distilled water of sterilizing
It is secondary；
3) under constant temperature non-illuminated conditions, with fluorine pyridine ketone solution processed soybeans seed 12-24h；
4) by step 3) soya seeds after processing are planted in soil, and cultivation temperature is 20-25 DEG C, during germination, fixed
Shi Dingliang sprays appropriate running water, and seed is sprouted through 2~10d.
Further, the mass concentration of pasteurization liquid is 5%-15%, and soak time is 5-15 minutes.
Further, the concentration of fluorine pyridine ketone solution is 0.05-0.15 μm of ol/L；Fluorine pyridine ketone solution treatment temperature is 20-25
℃。
Compared with prior art, the present invention can be obtained including following technique effect：
1) 0.05-0.15 μm of ol/L of the present invention fluorine pyridine ketone solution can significantly improve sprouting for the lower soya seeds of NaCl stress
Hair rate.
2) present invention can improve agricultural production efficiency on salt-soda soil.
Certainly, any product for implementing the present invention it is not absolutely required to while reaching all the above technique effect.
Brief description of the drawings
Accompanying drawing described herein is used for providing a further understanding of the present invention, constitutes the part of the present invention, this hair
Bright schematic description and description is used to explain the present invention, does not constitute inappropriate limitation of the present invention.In the accompanying drawings：
Fig. 1 is the seed sprouting that NaCl of the present invention suppresses different genotype soya bean；Wherein, full grains are chosen, size is equal
One, " ND-12 " (a) of no disease and pests harm and mechanical damage, " HD-19 " (b) soya seeds are respectively placed in that to be covered with two layers of middling speed qualitative
In the 9cm culture dishes of filter paper, cultivated under illumination condition；NaCl concentration is 150mmol/L, and control is used as using ultra-pure water
(CK)；Left side is followed successively by the photo of the germination rate difference maximum time point under illumination condition of the same race；Right side is followed successively by light of the same race
Germination rate statistical chart according under the conditions of；Scale=10mm；(c) it is respectively ND-12, HD-19 long in the root that 84h is measured；And fresh weight；
(d) it is respectively fresh weight that ND-12, HD-19 are measured in 84h.
Fig. 2 is the seed sprouting that NaCl of the present invention suppresses black soya bean；Wherein, full grains are chosen, size is homogeneous, no disease and pests harm
And " C-103 " (a) soya seeds of mechanical damage, it is respectively placed in and is covered with the 9cm culture dishes of two layers of middling speed qualitative filter paper, exists
Cultivated under illumination condition；NaCl concentration is 150mmol/L, and control (CK) is used as using ultra-pure water；Left side is under illumination condition
The photo of germination rate difference maximum time point；Right side is the germination rate statistical chart under illumination condition；Scale=10mm；(b), (c)
Respectively C-103 is in the 84h root length measured and fresh weight；
Fig. 3 is various concentrations NaCl solution suppression " the southern Germination of Soybean Seed of beans 12 " under illumination condition of the present invention；Wherein, select
Full grains are taken, size is homogeneous, " ND-12 " soya seeds of no disease and pests harm and mechanical damage are placed in that to be covered with two layers of middling speed qualitative
In the 9cm culture dishes of filter paper, cultivated respectively under 100mM/L, 150mM/L and 200mM/L NaCl solution；Made using ultra-pure water
For control (CK)；Left side is followed successively by the photo of the germination rate difference maximum time point under three kinds of concentration；Right side is in three kinds of concentration
Under the conditions of germination rate statistical chart.Scale=10mm.
Fig. 4 is NaCl increases Soybean Seeds Germination indices content of the present invention；Wherein, it is experiment with " ND-12 "
Material, takes the soya seeds of 24h, 48h, 72h and 96h control and the processing of 150mM/L NaCl solutions, respectively liquid feeding nitrogen respectively
Grinding, adds different reagents according to different enzymes are surveyed, centrifuges, and utilizes the different enzyme activity of spectrophotometric determination and mda content；(a),
(b), (c) unit U/g；(d) unit nmol/g；
Fig. 5 is the expression of ABA response genes in NaCl up-regulations Soybean Seeds Germination of the present invention；Wherein, with " ND-12 "
For experiment material, the soya seeds of 0h, 3h, 6h and 9h control and the processing of 200mmol/L NaCl solutions are taken respectively, extract total
MRNA, reverse transcription is carried out after removing genomic DNA, and ABA biosynthesis relative enzyme genes are determined using real time fluorescent quantitative technology
GmNCED9, GmNCED5 (a) and immediate early genes GmABI4, GmABI5 (b) relative expression quantity；
Fig. 6 be FL of the present invention alleviate NaCl to " the southern Germination of Soybean Seed of beans 12 " inhibition, wherein, FL can promote
" seed of southern beans 12 " is sprouted, and alleviates NaCl to " the southern Germination of Soybean Seed of beans 12 " inhibition；By " the southern soya seeds of beans 12 "
It is placed under illumination condition and cultivates, FL concentration is 0.1 μm of ol/L, using equivalent ultra-pure water as control；Left side is germination rate statistical chart
(a)；Right side is the long statistical chart (b) of root.* represent in 0.05 flat significant difference.
Embodiment
Describe embodiments of the present invention in detail below in conjunction with embodiment, thereby to the present invention how application technology hand
Section can fully understand and implement according to this to solve technical problem and reach the implementation process of technology effect.
The method that the present invention provides a kind of lower Germination of Soybean Seed of raising NaCl stress, comprises the following steps：
1) Seeds preprocess：Choose selection size after natural air drying homogeneous, full grains, the soya seeds of no disease and pests harm,
It is standby；
2) soya seeds surface sterilization, 5-15 points of medicining liquid dipping are carried out by 5%-15% pasteurizations liquid of mass concentration
Clock, is then cleaned 4 times with the distilled water of sterilizing；
3) under 20-25 DEG C of constant temperature non-illuminated conditions, with 0.05-0.15 μm of ol/L fluorine pyridine ketone solution processed soybeans seed
12-24h；
4) by step 3) soya seeds after processing are planted in soil, and cultivation temperature is 20-25 DEG C, during germination, fixed
Shi Dingliang sprays appropriate running water, and seed is sprouted through 2~10d.
Embodiment 1
A kind of method of the lower Germination of Soybean Seed of raising NaCl stress, comprises the following steps：Choose and chosen after natural air drying
Size is homogeneous, full grains, the soya seeds of no disease and pests harm, standby；Soya seeds surface is carried out with 10% pasteurization liquid to disappear
Then poison, medicining liquid dipping 10 minutes is cleaned 4 times with the distilled water of sterilizing；With 0.1 μm of ol/L fluorine pyridine ketone solution processed soybeans
Seed 12h, running water flowing water rinses 5min, and blotting paper blots surface moisture, is uniformly spread out on clean filter paper and dries to original
Dry seedses weight；Soya seeds after processing are planted in soil, cultivation temperature is 25 DEG C, during germination, is sprayed at regular time and quantity
Appropriate running water is spilt, seed is sprouted through 2~10d.
Embodiment 2
A kind of method of the lower Germination of Soybean Seed of raising NaCl stress, comprises the following steps：Choose and chosen after natural air drying
Size is homogeneous, full grains, the soya seeds of no disease and pests harm, standby；Soybean Species are carried out by 5% pasteurization liquid of mass concentration
Then sub- surface sterilization, medicining liquid dipping 15 minutes is cleaned 4 times with the distilled water of sterilizing；In under 20 DEG C of constant temperature non-illuminated conditions,
With 0.15 μm of ol/L fluorine pyridine ketone solution processed soybeans seed 12h, take out, running water flowing water rinses 5min, blotting paper blots table
Face moisture, is uniformly spread out on clean filter paper and dries to original dry seedses weight；Soya seeds after processing are planted in soil
In, cultivation temperature is 20 DEG C, during germination, and appropriate running water is sprayed at regular time and quantity, and seed is sprouted through 2~10d.
Embodiment 3
A kind of method of the lower Germination of Soybean Seed of raising NaCl stress, comprises the following steps：Choose and chosen after natural air drying
Size is homogeneous, full grains, the soya seeds of no disease and pests harm, standby；Soybean is carried out by 15% pasteurization liquid of mass concentration
The surface of the seed is sterilized, medicining liquid dipping 5 minutes, is then cleaned 4 times with the distilled water of sterilizing；In 25 DEG C of constant temperature non-illuminated conditions
Under, with 0.05 μm of ol/L fluorine pyridine ketone solution processed soybeans seed 24h, take out, running water flowing water rinses 5min, blotting paper is blotted
Surface moisture, is uniformly spread out on clean filter paper and dries to original dry seedses weight；Soya seeds after processing are planted in soil
In earth, cultivation temperature is 25 DEG C, during germination, and appropriate running water is sprayed at regular time and quantity, and seed is sprouted through 2~10d.
Illustrate the technique effect of the present invention with reference to specific experimentation：
1. experiment material and method
1.1 experiment materials
Using the soybean varieties (material) of three different genetic backgrounds：" southern beans -12 (ND-12) ", the " (HD- of He beans -19
" and " C-103 " 19)." southern beans -12 " are the main yellow soybean cultivars in In Southwest China, and " He beans -19 " are China of China
East and the main yellow soybean cultivars in North China, " C-103 " is black soybean cultivars.ND-12 and C-103 are four
The unified plantation in river agriculture university research and teaching farm (Sichuan, Yaan), harvest and storage；HD-19 is in Heze City, Shandong Province plantation, receipts
Obtain.Size is chosen after natural air drying homogeneous, full grains, it is related real that the soya seeds without obvious pest and disease damage carry out seed sprouting etc.
Test.
1.2 instruments, reagent
Instrument used in experiment mainly includes reverse transcription PCR instrument (the C100TM Thermai of Bio-RAD productions
Cycler), qPCR amplification instruments (the Two ColorReal-Time PCR Dtection of MyiQ 2 of Bio-RAD companies production
System), illumination box (25 DEG C of constant temperature, 2430LX).
Kit used in reverse transcription is ShiJi Co., Ltd from health, and kit used in qPCR comes from Vazyme.It is real
Primer used in testing is synthesized by giving birth to work (Shanghai).
Primer sequence in 1.3 experiments
The primer that table 1. is used in testing
1.4 experimental methods
1.4.1 culture dish filter paper cultivation
This culture experiment is carried out in illumination box, and specific incubation step is as follows：
1. Seeds preprocess：Choose uniform in size, the full soya seeds of health.
2. culture dish cultivates label and processing design：For the examination a diameter of 9cm of culture dish, first in clean glass culture dish
Double-layer filter paper is added, different disposal is done according to different experiments purpose.And it is labelled on culture dish respectively, by every 20 of seed
Uniformly it is put into the culture dish for being covered with filter paper, is compared with distilling water process.It is placed in (25 DEG C) constant temperature no light incubator and sends out
Bud.
3. observed and recorded：Seven time points of 12h, 24h, 36h, 48h, 60h, 72h and 84h are observed and record each training respectively
Support the sprouting number of seed in ware.
1.4.2 hot phenol method extracts plant total serum IgE
The molecular mechanism of inhibitory action is played to soybean germination for research NaCl, NaCl solution processing procedure need to be further probed into
The expression at gene tetra- time points of 0h, 3h, 6h and 9h after treatment of middle regulation seed ABA, GA content.Process step is such as
Under：1. Seeds preprocess.Choose uniform in size, the full soya seeds of health.With 150mM NaCl solution processed soybeans kind
Son；2. sampling：Sampled respectively four time points, 8 seeds are taken every time, are stored in liquid nitrogen；3. carried out with hot phenol method
The extraction of soybean total serum IgE.The sample of collection is ground rapidly in liquid nitrogen, moved into centrifuge tube, it is 65 DEG C of PH to add preheating
=6.0 phenol/chloroform and RNA extracts (50mM Tris-HCl, pH 6.0；10mM EDTA；2%SDS；100mM LiCl)
Isometric mixed liquor, fully shaking, and it is uniform to be vortexed.Rapid to put 4 DEG C, 12000r centrifugation 15min take supernatant, and add with it is upper
Clear isometric phenol/chloroform, is repeated after extracting 2-3 times, takes upper strata aqueous phase to add 1ml LiCl (4M), -80 DEG C stand 2 hours
More than, thawed after taking-up, remove supernatant, added after 75% ethanol washing precipitation, 12000r, 4 DEG C of centrifugation 5min, outwell ethanol,
Make in sterile environment after the alcohol volatilization completely of residual, add 20 μ L ddH2O, stayed overnight in 4 DEG C of refrigerator dissolvings.
1.4.3 reverse transcription
The synthesis HiFiscript Quick gDNA of DNA removals and the chain of genome cDNA first before reverse transcription
Removal CDNA Kit HiFiScript kits.First, it is to remove the DNA in extracted RNA, with reference to kit
Remove the reaction system sample-adding of genomic DNA.Plus after good medicine, the concussion that is vortexed is mixed, of short duration centrifugation, 42 DEG C of incubation 2min, instead
It is placed on ice after should terminating.
Reverse transcription reaction, the RNA and reverse transcription reaction system that remove DNA is well mixed on ice, of short duration centrifugation,
37 DEG C of incubations 15min, 85 DEG C of incubation 5s in PCR instrument, after reaction terminates, of short duration centrifugation is placed in cooled on ice.Reverse transcription product class
It is directly used in PCR and quantitative fluorescent PCR.
1.4.4 qRT-PCR
QRT-PCR reaction system is made as the AceQTMQpcr SYBR Green Master Mix's of VazymeTM companies
Configured with specification
Response procedures are as follows after improving：Heating period, 95 DEG C of 5min；Cycle stage, 95 DEG C of 10s, 55 DEG C of 30s, 40 are followed
Ring；Dissolution phase, 95 DEG C of 15s, 60 DEG C of 60s, 95 DEG C of 15s.
2. experimental result and analysis
2.1 Germination of Soybean Seed are tested
2.1.1NaCl the influence sprouted to different genotype soybean seed is handled
Using ultra-pure water as control, with 150mM NaCl solution handle different cultivars soya seeds (ND-12, C-103 and
HD-19), cultivated in illumination box.It was found that the soya seeds of each lower genotype of 150mM NaCl solution processing are sprouted
Hair rate is significantly lower than control group.Experiment is repeated several times, the NaCl solution for as a result showing 150mM is handled to different genotype
Obvious inhibitory action (Fig. 1) is played in the sprouting of soya seeds.It can be seen that by the root length and fresh weight data of different genotype soybean,
Compared with the control, Soybean Root length and fresh weight after NaCl solution is treated are significantly reduced.Carried on the back with soya bean (ND-12, HD-19) heredity
Scape differs the Data Representation of larger black soya bean (C-103) germination rate, fresh weight and root long side under 150mM NaCl solution processing
Go out the trend (Fig. 2) similar to soya bean (ND-12, HD-19).
After the difference between excluding genotype, NaCl solution still has obvious inhibitory action to the sprouting of soya seeds.Table
This bright depression effect generally existing, and it is unrelated with soybean heredity background.
2.1.2 influence of the NaCl solution of various concentrations to Germination of Soybean Seed
In order to further determine that inhibitory action of the NaCl solution to Germination of Soybean Seed, this experiment increase salinity 0mM,
100mM, 150mM, 200mM are handled soya seeds, and observational study various concentrations NaCl solution processed soybeans seed is sprouted
Heat condition.Experiment shows, with the increase of NaCl solution concentration, and its inhibition level to soybean germination also improves (Fig. 3) therewith.
Figure 3 above shows that sprouting of the NaCl solution to soya seeds is inhibited.Over time, with compareing
Compare, the Germination of Soybean Seed rate after NaCl solution is treated is substantially reduced, and in 48h, control group soybean germination rate is
The soybean germination rate of 93.33%, 100mM NaCl solution processing is the soybean germination that 68.33%, 150mM NaCl solutions are handled
Rate is that the soybean germination rate of 40.00%, 200mM NaCl solutions processing is 11.67%, in notable downward trend.
2.2 resistance relative physiologic index CAT, SOD, POD and MDA measure
From Fig. 4 a, 4c, in the 24h-96h after soya seeds imbibition, compared with the control, after NaCl solution processing
Catalase (CAT) and superoxide dismutase (SOD) content are dramatically increased in soybean.In Fig. 4 b, in soya seeds imbibition
In 72h-96h afterwards, compared with the control, peroxidase (POD) content is dramatically increased in the soybean after NaCl solution processing.
In Fig. 4 d, because the content of MDA (MDA) is changed by enzyme effect, therefore the 96h after soya seeds imbibition or so, NaCl
MDA (MDA) is significantly higher than control in soybean after solution processing.
The expression of the controlling gene of seed ABA contents is influenceed in 2.3 NaCl solution processing procedures
With the success that arabidopsis gene group is sequenced, arabidopsis has turned into the ideal material that research seed sprouts mechanism.It is logical
Cross the discovery of the research to ten various mutations bodies of arabidopsis.Many sproutings of the signaling molecule particularly ABA and GA to seed are played
Important effect, ABA is relevant with the dormancy of seed, and has certain effect to the sprouting of seed.This experiment is being determined
After sprouting of the NaCl solution to soya seeds has a significant impact, the successful experience of arabidopsis research is used for reference, is further probed into
The correlation that NaCl solution influences on ABA expression quantity.
ABA plays an important roll in plant germination is suppressed, it has now been found that abscisic acid is relevant with seed dormancy, molten in NaCl
Regulate and control gene expression under salt stress caused by liquid.In recent years to the identification for the gene for encoding ABA biosynthesis key enzymes, disclose
The biological approach of ABA synthesis.The presence of gene family simultaneously and their respective effects illustrate the mechanism of ABA biosynthesis.
Once ABA hormone-contents level is raised, signal transduction mechanism is activated, stimulated gene expression.Therefore this experimental design passes through
Detect that the soya seeds after the processing of 200mM NaCl solutions in tetra- time points of 0h, 3h, 6h and 9h, regulate and control seed and sprouted after treatment
Expressions of gene GmNCED9, GmNCED5, GmABI4, the GmABI5 of hair process ABA contents in seed.
2.3.1 abscisic acid (ABA) synthesis related gene transcription research
The committed step of ABA biosynthesis be 9- it is cis-neoxanthin dioxygenase (9-cis-epoxycarotenoid
Dioxygenase, NCED) further catalysis 9- it is cis-neoxanthin (9-cis-neoxanthin) produce xanthoxin
(xanthoxin), xanthoxin is entered in cytoplasm with a kind of mechanism not yet understood, is converted into the presence of ABA2/SDR1
Abscisic acid aldehyde, finally, abscisic acid aldehyde are generated in the presence of abscisic acid aldehyde oxidase (ABA-aldehyde oxidase, AAO)
The ABA of activity.Therefore, ABA synthesis is controlled by NCED gene families.
Detected by the qPCR to the controlling gene (NCED5, NCED9) in germination process on ABA biosynthesis, with
ND-12 is experiment material, and the GmNCED genes for further probing into regulation seed ABA contents in NaCl solution processing procedure exist
The expression (Fig. 5 a) at tetra- time points of 0h, 3h, 6h and 9h after 200mM NaCl solution processing.
It can be known by Fig. 5 a, after treatment in 6 hours, the ND-12 soya seeds that NaCl solution is handled compared with control group
GmNCED5 expression quantity is dramatically increased, and top occurs in GmNCED5 expression quantity the 3rd hour after treatment,
GmNCED5 expression quantity is close to twice of expression quantity after the processing of the control group aqueous solution.The GmNCED9's of ND-12 soya seeds
There is top in after treatment 3 hours in expression quantity, and GmNCED9 expression quantity is higher than during 3h to 9h after treatment
Control group expression quantity.Known NCED plays important regulating and controlling effect in ABA biosynthesis, shows relative to control group
Speech, the ABA contents levels in soya seeds, which have, after treatment substantially increases.
2.3.2 abscisic acid (ABA) signal transduction related gene transcription analysis
ABA signal paths react as a complicated phosphorylation cascade, are related to a variety of positive and negative regulatory factors.By right
The molecule genetics research of arabidopsis, has identified the component of a variety of ABA signal transduction paths, and at present, what is found in plant is more
Plant the signal transduction that PP2C class phosphatases both participate in ABA.So far, it has been found that the member of arabidopsis PP2C families is such as
ABI1, ABI2, HAB1, HAG3 and PP2CA etc., they are ABA signal pathway regulatory factors.They are by changing ABA signals
Strong and weak regulation and control plant stress response, so as to influence ABA signal transductions.
By to being detected in germination process to GmABI4, GmABI5 qPCR, further probing into NaCl solution processing procedure
The table at these genes of middle regulation seed ABA contents tetra- time points of 0h, 3h, 6h and 9h after 200mM NaCl solution processing
Up to situation (Fig. 5 b)
Knowable to Fig. 5 b, soya seeds, GmABI4, GmABI5 expression quantity is at place after 200mM NaCl solution processing
Obviously higher than control group in 9 hours after reason.And, GmABI4, GmABI5 expression quantity after treatment three hours when there is highest
Peak, it is then on a declining curve.These known genes play important regulating and controlling effect, 200mM NaCl solution in ABA signal transductions
After processing, these gene great expressions reach the effect of the sprouting for suppressing soybean, finally by promoting ABA biosynthesis
Show as salt stress phenomenon.
2.4 fluorine pyridine ketone (Fluridone, FL) are to the lower recovery for sprouting delay of NaCl processing
Three groups of processing of Setup Experiments (ultra-pure water, the processing of 150mMNaCl solution, 150mM NaCl+0.1 μm ol/L FL) exist
Cultivated in illumination box.The ND-12 sprouting situations (Fig. 6) of observational study ABA synthetic inhibitors FL processing.
From Fig. 6 a, the processing containing FL can alleviate the inhibition that NaCl sprouts to ND-12.During 24h, 150mM
NaCl+0.1 μm of ol/L FL processing ND-12 germination rate is higher by 26.25% than the germination rate that same concentration NaCl solution handles ND-12.
In Fig. 6 b, root appearance of the 150mM NaCl+0.1 μm ol/L FL processing ND-12 roots length with handling ND-12 with concentration NaCl solution
Than not notable, it is contemplated that the main component of herbicide is FL in agricultural production, may suppress the growth of plant, although low concentration
FL can promote Germination of Soybean Seed, but facilitation is had no to root length and fresh weight isophenous data.
3. discuss
Soybean Seeds Germination is the process of a hormone metabolism and various relevant enzymes synergy.Different processing sides
Method, has differences to the influence that seed is sprouted, and the factor for promoting seed to sprout may sprout institute with kind of a transdermally, activated seed
Required enzyme, plant hormone metabolism etc. are relevant.Change more than the current research to soybean germination in heavy metal ion, chemicals etc.
In terms of the physical factor such as factor aspect and temperature, moisture, and for soybean its Endogenous Hormone Contents in Vitro under condition of salt stress
Research is not yet seen in report.This experiment elaborates soybean under salt stress by strengthening ABA synthesis and signal in soya seeds
Intensity, suppresses the mechanism of Germination of Soybean Seed.And regulate and control seed and sprout related another important hormone gibberellin due to correlation
Signal path is more complicated, and this research not yet draws corresponding conclusion.This would is that this experiment group following research topic it
One.
With the increase of NaCl concentration for the treatment of, the inhibition of soybean germination is more obvious.In active oxygen relevant enzyme content,
POD, SOD, CAT enzyme content are substantially less than control group；MDA MDA content Initial changes are not obvious in cell, this be because
Changed for MDA content by enzyme effect, 48h no significant differences compared with the control before institute, MDA contents show after sprouting 72 hours
Write and be higher than control group.On molecular level, the up-regulation of ABA synthetic gene GmNCED5 and GmNCED9 expression, Transcription-related gene
GmABI4, GmABI5 expression are raised.Soybean germination experiment is then carried out jointly with ABA inhibitor FL and NaCl, is as a result demonstrate,proved
Bright NaCl regulation and control ABA synthesis and the sprouting for transcribing and suppressing soya seeds.
This research is proved NaCl regulation and control ABA synthesis and transcribes and suppress the sprouting of soya seeds, and is tested by FL
It was found that low concentration FL can alleviate suppression of the NaCl to Germination of Soybean Seed.
Some preferred embodiments of invention have shown and described in described above, but as previously described, it should be understood that invention is not
Form disclosed herein is confined to, the exclusion to other embodiment is not to be taken as, and available for various other combinations, modification
And environment, and can be carried out in invention contemplated scope described herein by the technology or knowledge of above-mentioned teaching or association area
Change., then all should be in the appended power of invention and the change and change that those skilled in the art are carried out do not depart from the spirit and scope of invention
In the protection domain that profit is required.
SEQUENCE LISTING
<110>Sichuan Agricultural University
<120>A kind of method of the lower Germination of Soybean Seed of raising NaCl stress
<130> 2016
<160> 18
<170> PatentIn version 3.3
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Claims (3)
1. a kind of method of the lower Germination of Soybean Seed of raising NaCl stress, it is characterised in that comprise the following steps：
1) Seeds preprocess：Selection size is homogeneous after selection natural air drying, and full grains, the soya seeds of no disease and pests harm are standby；
2) soya seeds surface sterilization is carried out with pasteurization liquid, then medicining liquid dipping is cleaned 4 times with the distilled water of sterilizing；
3) under constant temperature non-illuminated conditions, with fluorine pyridine ketone solution processed soybeans seed 12-24h；
4) by step 3) soya seeds after processing are planted in soil, and cultivation temperature is 20-25 DEG C, during germination, and timing is fixed
The appropriate running water of amount sprinkling, seed is sprouted through 2~10d.
2. the method for the lower Germination of Soybean Seed of raising NaCl stress according to claim 1, it is characterised in that the Pasteur
The mass concentration of thimerosal is 5%-15%, and soak time is 5-15 minutes.
3. the method for the lower Germination of Soybean Seed of raising NaCl stress according to claim 1, it is characterised in that the fluorine pyridine
The concentration of ketone solution is 0.05-0.15 μm of ol/L；Fluorine pyridine ketone solution treatment temperature is 20-25 DEG C.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108513760A (en) * | 2018-04-19 | 2018-09-11 | 四川农业大学 | A method of promoting Germination of Soybean Seed under Submergence stress |
| CN113207884A (en) * | 2021-04-15 | 2021-08-06 | 湖南农业大学 | Application of fluazinone as strigolactone inhibitor |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105659975A (en) * | 2016-01-22 | 2016-06-15 | 曾少兰 | Method for increasing germination rate of soybean seeds |
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2017
- 2017-03-23 CN CN201710179195.4A patent/CN106941810A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105659975A (en) * | 2016-01-22 | 2016-06-15 | 曾少兰 | Method for increasing germination rate of soybean seeds |
Non-Patent Citations (3)
| Title |
|---|
| RUNQIANG YANG 等: "Effects of ABA and CaCl2 on GABA accumulation in fava bean germinating under hypoxia-NaCl stress", 《BIOSCIENCE，BIOTECHNOLOGY，AND BIOCHEMISTRY》 * |
| 董钻: "《大豆产量生理》", 30 November 2000, 中国农业出版社 * |
| 魏开发 等: "ABA信号转运调节的基因表达与源库动力学分析", 《云南植物研究》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108513760A (en) * | 2018-04-19 | 2018-09-11 | 四川农业大学 | A method of promoting Germination of Soybean Seed under Submergence stress |
| CN113207884A (en) * | 2021-04-15 | 2021-08-06 | 湖南农业大学 | Application of fluazinone as strigolactone inhibitor |
| CN113207884B (en) * | 2021-04-15 | 2022-06-21 | 湖南农业大学 | Application of fluazinone as strigolactone inhibitor |
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