CN108467868A - The application of soybean sucrose transporter important gene GmSWEET6 - Google Patents
The application of soybean sucrose transporter important gene GmSWEET6 Download PDFInfo
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Abstract
The present invention discloses a kind of application of soybean sucrose transporter important gene GmSWEET6, is related to plant genetic engineering and biotechnology.The present invention identifies the sucrose transporter gene GmSWEET6 of a mycorhiza induced expression by real time fluorescence quantifying PCR method in soybean, the regulation and control that the expression of the gene is infected by mycorrhizal fungi.Under mycorrhizal fungi inoculation condition, there is regulating plant growth and phosphorus to absorb by excess or interference expression GmSWEET6 in Transgenic soybean plants, there is significant impact to the beneficial symbiosis of soybean and arbuscular mycorrhizal fungi, this is to illustrating biological function of the SWEET genes in legume and mycorrhizal fungi symbiosis, to regulate and control the beneficial symbiosis important in inhibiting between plant and mycorrhizal fungi.
Description
Technical field
The present invention relates to plant genetic engineerings and biotechnology, and in particular to a kind of soybean sucrose transporter is important
The application of gene GmSWEET6.
Background technology
Higher plant is the autotroph being made of autotrophic organ and heterotrophism organ.Heterotrophism organ needs the light of autotrophic organ
Product is closed to maintain growth and development.The photoassimilates 80% synthesized in plant maturation blade etc. " source " organ all pass through plant
Skeleton transports root, flower, fruit etc. " library " organ.Sucrose is one of green plants photoassimilates, and most
Photosynthate trasport and the principal mode of distribution in higher plant body (Yang Shuan etc., 2005；Bai Xuemei etc., 2006).Sucrose is being planted
The mode of orientation transport and distribution not only regulates and controls the entire growth and development process of plant in strain, also determines the yield and product of crop
Matter.SWEET (Sugars Will Eventually be Exported Transporters) is as a kind of new sugar transport egg
In vain, have the function of arranging outside sucrose and monosaccharide, this may be the key point that sucrose is unloaded to the libraries such as root organ from bast.
There are numerous studies to show that SWEET genes participate in various physiological processes, including phloem loading, influence reproductive development, participation place
The disease resistance response and Stress responses of master-pathogen interaction and ion transport etc..It has been reported that it is relevant with rhizobium symbiosis
MtSWEET11；12 StSWEETs genes up-regulated expressions after Arbuscular Mycorrhizal Fungi in potato, this shows that SWEET turns
Fortune albumen may participate in the process that sucrose arranges sugar to outside yeast-like symbiont.
Before 400,000,000 5 thousand ten thousand years, arbuscular mycorrhizal fungi (Arbuscular Mycorrhiza Fungi, AMF) and vegetal inspired
At symbiosis (Wang et al., 2006).Now, there is about 80% or more terrestrial plant that can be formed with AM fungies
Cogeneration system (Smith et al., 2008).The spore of AM fungies is sprouted in the soil, and mycelia invaded plants root system is thin to cortex
In born of the same parents, clump branch structure is then formed.The outer mycelia of the root of AM fungi prosperities can reach the region that root system of plant cannot contact and inhale
Nutriment (especially phosphorus) is received, and plant is passed to by the mycorhiza approach of cogeneration system, to improve the nutrient effect of host
Rate.Moreover, the heavy metallic poisons such as aluminium, manganese can also be effectively relieved in the formation of cogeneration system, and raising plant is resisted abiotic
The ability of stress.As return, plant provides carbon source, including carbohydrate for AM fungies, for fungi clump branch, mycelia, spore and
Being normally carried out of the growth and development of the basic structures such as vesicle and life cycle (Varma et al., 1995；Read et al.,
1997).Therefore, in cogeneration system, sucrose transporter also necessarily plays vital effect.
Soybean is the important grain and oil crop in China.The soybean of countries in the world cultivation is all directly or indirectly to be propagated by China
(He Jinshang, 2009) to go out.Identify 53 GmSWEETs gene families member (Patil et altogether in soybean at present
al.,2015).Up to the present, in mycorrhizal plants, there are no finding to be responsible for that yeast-like symbiont will be discharged to outside sucrose, mycorhiza lures
SWEET transport proteins are led, and there are no the correlative studys of the SWEET transport proteins induced about mycorhiza in soybean.
Invention content
In order to overcome the disadvantages and deficiencies of the prior art, the purpose of the present invention is to provide a kind of soybean sucrose transporters
The application of important gene GmSWEET6.
The purpose of the invention is achieved by the following technical solution：
The present invention provides a kind of application of soybean sucrose transporter important gene GmSWEET6 in terms of Mycorrhizal Symbiosiss.
The nucleotides sequence of the soybean sucrose transporter important gene GmSWEET6 is classified as Glyma.04G198600 institutes
Show, the amino acid sequence of the protein of GmSWEET6 gene codes is shown in Glyma.04G198600.
By real time fluorescence quantifying PCR method, the sucrose that a mycorhiza induced expression is identified in soybean turns the present invention
Transport protein gene GmSWEET6, the regulation and control that the expression of the gene is infected by mycorrhizal fungi.Under mycorrhizal fungi inoculation condition, transgenosis
There is regulating plant growth and phosphorus to absorb by excess or interference expression GmSWEET6 in soybean plant strain, to regulation and control plant and mycorrhizal fungi
Between beneficial symbiosis be of great significance.
The present invention also provides a kind of applications of soybean sucrose transporter important gene GmSWEET6 in plant breeding, into
One step, the application in cultivating genetically modified plants.
The plant is dicotyledonous legume.
The dicotyledonous legume is soybean.
Gene GmSWEET6 and albumen provided by the invention can regulate and control the growth of the transfer-gen plant comprising it and phosphorus is inhaled
It receives.
The primer pair for expanding above-mentioned GmSWEET6 full length genes or its any segment belongs to the scope of protection of the present invention.
The present invention also provides the recombinant expression carriers containing above-mentioned GmSWEET6 genes, can use existing plant expression vector
Build the recombinant expression carrier containing GmSWEET6 genes.The plant expression vector includes double base agrobacterium vector etc., such as
PTF101.1 (Wang et al., 2009) or other derivative plant expression vectors.
The present invention also provides a kind of genetic engineering bacteriums, contain above-mentioned recombinant expression carrier.
The invention further relates to cells, and it includes the GmSWEET6 genes or recombinant expression carrier of the present invention.The cell can
To be plant cell, such as legume cell or microbial cell, such as bacterium or fungal cell.The cell can be with
It is separation, in vitro, part culture or that be plant.
The invention further relates to plant or plant parts, vegetable material, and vegetable seeds, it includes the cells of the present invention.Institute
It can be legume, such as Kidney bean and soybean to state plant, can also be other plants, such as monocotyledon such as rice, small
Wheat, barley, corn, sorghum, sugarcane, oat, rye etc. or other dicotyledons for example tobacco, sunflower, beet, capsicum,
Potato, tomato etc..Further relate to the transgenic seed from the plant.
The invention further relates to the method for production plant, this method includes：It is planted from the Plant cell regeneration transgenosis of the present invention
Object, or by plant and another plant hybridization of the present invention.
The invention further relates to the plants of the method for present invention production.
The present invention also provides a kind of soybean sucrose transporter important gene GmSWEET6 in regulation and control plant and mycorrhizal fungi
Application in being formed beneficial to symbiosis；Further, regulation and control plant and the application in mycorrhizal fungi symbiosis preparation are being prepared.
The plant is dicotyledonous legume.
The dicotyledonous legume is soybean.
It is beneficial altogether with mycorrhizal fungi in regulation and control plant that the invention further relates to the GmSWEET6 genes of the present invention or recombinant vector
Raw relationship formed in purposes, including prepare transgenosis plant and prepare to change plant and formed with mycorrhizal fungi and closed beneficial to symbiosis
The preparation of system.
The invention further relates to regulation and control plants and mycorrhizal fungi to form the method beneficial to symbiosis, and this method includes preparing to contain
There are the GmSWEET6 genes of the present invention or the plant of recombinant vector.For example, the method may include the plant from the present invention is thin
Born of the same parents' regenerating plants or by plant and another plant hybridization of the present invention.
A preferred embodiment provided by the present invention is that said gene GmSWEET6 is imported soybean hypocotyl injection
In the root system of induction, transgenic line is obtained；Apparent become occurs for the growth of the transgenic line, including biomass and phosphorus content
Change.
The gene GmSWEET6 can import receptor soybean hypocotyl root system by the recombinant expression carrier.
The plant expression vector for carrying the gene GmSWEET6 of the present invention can be for example, by Agrobacterium-medialed transformation method
It is transformed into soybean hypocotyl root system.
The present invention has the following advantages and effects with respect to the prior art：
Though the SWEET gene families belonging to gene GmSWEET6 have been cloned and have reported in arabidopsis and clover etc., its
It is not known beneficial to the biological function in terms of symbiosis in legume and arbuscular mycorrhizal fungi.The gene GmSWEET6 of the present invention
There is significant impact to the beneficial symbiosis of soybean and arbuscular mycorrhizal fungi, this is to illustrating SWEET genes in legume and bacterium
Biological function in mycorrhiza fungi symbiosis, to regulate and control the beneficial symbiosis important in inhibiting between plant and mycorrhizal fungi.
Description of the drawings
Fig. 1 is reaction of the GmSWEETs family genes in root to AM fungal infections；Plant is selected using sand training experiment
Two different soybean genotype BD2 and BX10, in low-phosphorous (LP, 50 μM of KH2PO4) or high phosphorus (HP, 500 μM of KH2PO4)
It is harvested after under treatment conditions, not being inoculated with (NM) or Arbuscular Mycorrhizal Fungi Ri 45d.Ri：Rhizophagus
irregularis.a-h：Expression quantity of the GmSWEETs genes in root；The average value that data in figure repeat for four secondary pollutants ±
SE.Asterisk (*) indicates under same gene type and the processing of same phosphorus that plant connects bacterium and do not connect significant difference between bacterium.
Fig. 2 is that the GUS expression of GmSWEET6 promoters driving and the common location of mycorhiza structure are analyzed；Wherein, Fig. 2 a are
It is the fluorescing matter of the mycorhiza structure after WGA488 is dyed that GmSWEET6 promoters, which drive the expression of GUS, Fig. 2 b,.
Fig. 3 is the Subcellular Localization of GmSWEET6；First row is the Tobacco Epidermis for converting empty carrier in figure, tightly
Followed by the tobacco cell of conversion GFP-GmSWEET6 carriers, figure is observation shooting GFP and film positioning under Laser Scanning Confocal Microscope
The fluorescent co-location of Marker mCherry.
Fig. 4 is excessive and the expression quantity detection of interference expression GmSWEET6 transfer-gen plants.Using hypocotyl injecting method,
Over-express vector GmSWEET6-OX-pTF101.1s and interference vector GmSWEET6-pFGC5941 are transferred to phosphorus efficiency kind
In YC03-3, in low-phosphorous (50 μM of KH2PO4) under conditions of, it is not inoculated with (NM) or inoculation clump branch bacterium (Ri) is handled.A, b are on the ground
The gene expression amount data in portion；C, d are root data.Wherein, CK1 indicates the control for being transferred to pTF101.1s zero loads；6-OX is indicated
Overexpression GmSWEET6 transfer-gen plants；CK2 indicates the control for being transferred to pFGC5941 zero loads；6-RNAi indicates interference expression
GmSWEET6 transfer-gen plants.Data are the average value and standard error that five secondary pollutants repeat in figure.* it indicates same and connects bacterium processing
Significant difference (P between lower difference transgenic line<0.05).
Fig. 5 is excessive and interference expression GmSWEET6 is to Soybean transgenic plant phosphorus content (a, b) and plant strain growth situation
The influence of (c, d).Using hypocotyl injecting method, by over-express vector GmSWEET6-OX-pTF101.1s and interference vector
GmSWEET6-pFGC5941 is transferred in phosphorus efficiency kind YC03-3, in low-phosphorous (50 μM of KH2PO4) under conditions of, it is not inoculated with (NM)
Or inoculation clump branch bacterium (Ri) is handled.A, b are root phosphorus content；C, d are root biomass.Wherein, CK1 expressions are transferred to
The control of pTF101.1s zero loads；6-OX indicates overexpression GmSWEET6 transfer-gen plants；CK2 expressions are transferred to pFGC5941 skies
The control of load；6-RNAi indicates interference expression GmSWEET6 transfer-gen plants.Data are being averaged for five secondary pollutants repetition in figure
Value and standard error.* it indicates same and meets under bacterium processing significant difference (P between different transgenic lines<0.05).
Specific implementation mode
Present invention will now be described in further detail with reference to the embodiments and the accompanying drawings, but embodiments of the present invention are unlimited
In this.
Test method used in following embodiments is conventional method unless otherwise specified；Used material, examination
Agent etc., unless otherwise specified, for the reagent and material commercially obtained.
Embodiment 1
Expression pattern analysis of the GmSWEETs gene families in soybean root system：Soybean SWEET gene families share 53
Member (Patil et al., 2015), according to the affinity in evolution, screening is transported with sucrose and microbe symbiotic is relevant
Third props up 28 members as candidate gene, then by quantitative PCR technique, determines GmSWEETs family member's genes in difference
Soybean varieties, the different expressions connect under bacterium processing and different P levels.
Using 2 soybean varieties, phosphorus efficiency Brazil 10 (BX10) and inefficient local 2 (BD2) of phosphorus are arranged 2 AMF and are inoculated with
Processing is not inoculated with (NM) and Inoculation of Arbuscular Mycorrhizal Fungi Rhizophagus irregularis (Ri) respectively.Two phosphorus of setting are handled, and 50
μM KH2PO4As low-phosphorous (LP) and 500 μM of KH2PO4As high phosphorus (HP), the K of low-phosphorous processing+Use K2SO4Polishing.Using 1/
2Hoagland nutrient solution sands cultivate plant.It selects the red plastic of 2kg specifications to justify basin, quartz sand 1.6kg is packed into per basin, it is acid
Native 0.2kg, sand is through 121 DEG C of high pressure sterilization 40min, and cycle is twice.Corresponding AMF microbial inoculums or-AMF simulation inoculations are added when filling basin
Object 0.2kg (10%) is soaked with the corresponding nutrient solutions of 150mL after complete mixing, and soya seeds are gone out with the method for chlorine hot air sterilization
Then bacterium mends the corresponding nutrient solutions of 100mL again per basin 3 seeds of program request per basin.Thinning after emerging 1 week retains 1 plant per basin.
The 1/2Hoagland nutrient solutions for pouring corresponding (LP/HP) weekly are primary.Each processing is arranged 4 biology and repeats.Routine Management, 7
It is harvested after week, extracts overground part and root RNA, reverse transcription further uses the expression mould of quantitative PCR detection GmSWEETs at cDNA
Formula.The house-keeping gene EF-1a of soybean is as internal reference.Primer for quantitative PCR detection gene expression amount is respectively：
The primer of soybean EF-1a genes is：
EF-1a F：5′-TGAACCACCCTGGTCAGATT-3′(SEQ ID NO:1)
EF-1a R：5′-TCCAGCATCACCATTCTTCA-3′(SEQ ID NO:2)
The primer of GmSWEET6 genes is：
GmSWEET6F：5′-CGTTCGGGAGCGTAACATAG-3′(SEQ ID NO:3)
GmSWEET6R：5′-TCGGACCAAAAGCGTAGAGT-3′(SEQ ID NO:4)
Other GmSWEETs gene quantification primer sequences are shown in the paper of (2015) Patil etc..
QPCR reaction systems：10μLPowerUpTM Green Master Mix, 0.6 μ L Primer R,
0.6 μ L Primer F, 2 μ L cDNA, ddH2O Up to 20μL。
Wherein cDNA is used to test after diluting 20 times, takes cDNA stostes at random from each different experiment process, makes
Mark song ensures that each handles at least one and is used for mixing mark song, and mark song is at 5 times of gradient dilution.
PCR reaction conditions are 95 DEG C and are denaturalized 1 minute, are then denaturalized 15 seconds for 95 DEG C, 60 DEG C are annealed and extended 60 seconds, carry out 40
Secondary cycle.
Data analysis：Suitable threshold value is selected in ABI Step OnePlus biosystems makes standard curve
R2Reach 1, E values as possible between 85%~110%, ABI Step One Plus biosystems softwares are according to standard curve
Automatically the CT values of target gene and reference gene are calculated and obtains an expression quantity, the concentration of target gene and the table of reference gene
Ratio up to amount is the relative expression quantity of target gene.In order to ensure the accuracy of experimental result, each processing is at least arranged 4
A biology repeats, it is ensured that reaction tendency is consistent.The results are shown in Figure 1, and (unlisted gene is by low-phosphorous and/or mycorhiza in figure
Induction is not expressed).
The result shows that：GmSWEET6 is expressed by mycorhiza induced strong, especially under the conditions of low-phosphorous.
As a result imply that GmSWEET6 may play an important role in the beneficial symbiosis of plant and mycorrhizal fungi.
Embodiment 2
1, the clone of GmSWEET6 genes：Using soybean YC03-3 leaf portions cDNA as masterplate, with upstream specific primer 5 '-
ATCGCCCGGGATGTCGTCCCACAGTCATCTAA-3′(SEQ ID NO:And downstream special primer 5 ' -5)
ATCGCCCCGGGTCAAACTTCGCAACTGATCACCC-3′(SEQ ID NO:6) the ORF overall length sequences of GmSWEET6 genes are expanded
864bp is arranged, and comparison is sequenced, GmSWEET6 coded sequences is obtained and sees that Glyma.04G198600, corresponding protein sequence are shown in
Glyma.04G198600。
2, GmSWEET6 gene promoters clone, vector construction and tissue expression positioning analysis：Promoter Analysis expression carries
The structure of body：Conventionally, the leaves genomic DNA for extracting soybean YC03-3 genotype, with soybean leaf portion genomic DNA
For template, with 5 '-CTATGACATGATTAC of upstream specific primerGAATTCCCACCTTGTTATACCTCATT-3′(SEQ ID
NO:And 5 '-GACTGACCTACCCGG of downstream special primer 7)GGATCCGGAATTTCTCTCTCTCTCTCT-3′(SEQ ID
NO:8) expand GmSWEET6 promoter 2176bp segments, PCR fragment recycling after, by EcoR I and BamH I to purpose carrier into
After row double digestion, GmSWEET6 promoter genes are connected to purpose carrier pTF102 by the method for one-step cloning.Conversion is big
Enterobacteria DH10B is sequenced errorless rear conversion Agrobacterium tumefaciems EHA101, is used for the Agrobacterium tumefaciens mediated whole strain of soybean cotyledon node
Conversion (Wang et al., 2009).
The whole strain transformation plant of promoter is selected using sand culture test for GmSWEET6 tissue expression positioning analysis
ProGmSWEET6 is Ri, low-phosphorous 50 μM of KH for examination network of roots mycorrhizal fungi2PO4Nutrient solution.The K of low-phosphorous processing+Use K2SO4It mends
Together.10 biology of setting repeat.Routine Management, florescence collect fresh root sample, clean, and carry out GUS vital stainings.So
The root for dying blue is cut into small root segment afterwards, it is longitudinal sectional after being embedded with low melting point agar, with WGA488 to the Infection structure of AM fungies
It is dyed, is then observed and taken pictures at light field and 488nm wavelength respectively with fluorescence microscope.As a result see Fig. 2.The result shows that：
GmSWEET6 is induced to express in the clump branch structure of mycorhiza.
Embodiment 3
The structure of subcellular localization experimental vehicle：Using soybean YC03-3 leaf portions cDNA as masterplate, with upstream specific primer 5 '-
GGGGacaagtttgtacaaaaaagcaggcttcATGTCGTCCCACAGTCATCTAA-3′(SEQ ID NO:9) and downstream is special
Different primer 5 '-GGGGaccactttgtacaagaaagctgggtcTCAAACTTCGCAACTGATCACCC-3 ' (SEQ ID NO:
10) the ORF full length sequences 864bp of amplification GmSWEET6 genes, PCR recycle segment, by Gatway technologies by GmSWEET6 bases
Because being connected to purpose carrier pMDC43.Escherichia coli DH10B is converted, errorless rear conversion Agrobacterium GV3101 is sequenced, for carrying out
Subcellular localization is tested.
For the Subcellular Localization of GmSWEET6, using the method for Tobacco Epidermis instantaneous conversion.It first will fusion
Have the GV3101 bacterium solutions of GmSWEET6 genes and be transferred to film Marker plasmids 1008 GV3101 bacterium solutions shake bacterium stay overnight, then from
The heart, with including 10mM MES, 10mM MgCl2With the re-suspension liquid resuspended bacterium solution of 150 μM of AS.22~24 DEG C of 3~4h of light culture,
After the two is mixed in equal volume, in conversion to the tobacco leaf of 3~4 week old, after converting 3 days, in laser co-focusing 488nm
(GFP) and at 587nm (mCherry) fluorescence is observed.Experimental result is shown in Fig. 3.The result shows that：GmSWEET6 is located in plasma membrane and core
On film.
Embodiment 4
1, the structure of overexpression GmSWEET6 carriers：Using the leaf portion cDNA of soybean YC03-3 genotype as template, use
Swim 5 '-GCGAGCTCGGTA of special primerCCCGGGATGTCGTCCCACAGT CATCTAA-3′(SEQ ID NO:And downstream 11)
5 '-CTCTAGAGGAT of special primerCCCCGGGTCAAACTTCGCAACTGATCACCC-3′(SEQ ID NO:12) it expands
After PCR recycles segment, digestion is carried out by Sma I to purpose carrier by the ORF full length sequences 864bp of GmSWEET6 genes, will
GmSWEET6 genes are connected to purpose carrier pTF101.1s, obtain over-express vector GmSWEET6-OX-pTF101.1s.Conversion
Errorless rear transforming agrobacterium rhizogenes K599 is sequenced in Escherichia coli DH10B, the soybean hypocotyl injection mediated for agrobacterium rhizogenes
Conversion (Li et al., 2014).
2, interfere the structure of GmSWEET6 carriers：Using the leaf portion cDNA of soybean YC03-3 genotype as template, with upstream spy
Different 5 '-TTACCATGG of primerGGCGCGCCGATATCTATGTTACGCTCCC GA-3′(SEQ ID NO:13) and downstream is special
5 '-CATCGATTG of primerGGCGCGCCAACTT CGCAACTGATCACCCTTTC-3′(SEQ ID NO:14) amplification 390bp is special
Different target fragment, after carrying out digestion to purpose carrier using Asc I, with the method for one-step cloning by GmSWEET6 gene forward direction pieces
Section is connected to purpose carrier pFGC5941.Then upstream specific primer 5 '-is used again
GCAGGTATTTGGATCCAACTTCGCAACTGATCACCCTTTC-3′(SEQ ID NO:And downstream special primer 5 ' -15)
GACTCACCTAGGATCCGATATCTATGTTACGCTCCCGA-3′(SEQ ID NO:16) reversed segment is expanded, BamH is used
After I carries out digestion to purpose carrier, it includes that the purpose of positive segment carries to be connected to reversed segment with the method for one-step cloning
In body pFGC5941, interference vector GmS WEET6-pFGC5941 are obtained, convert Escherichia coli DH10B, errorless rear conversion is sequenced
Agrobacterium rhyzogenesK599, the soybean hypocotyl injection conversion (Li et al., 2014) mediated for agrobacterium rhizogenes.
3, the acquisition of Soybean transgenic plant：By the excess built and interference GmSWEET6 expression vector plasmids conversion
Into agrobacterium rhizogenes, the soybean hypocotyl injection conversion mediated using agrobacterium rhizogenes obtains transgenosis chimeric plant (Li et
Al., 2014), subsequent phenotypic evaluation uses transgenosis to be fitted into strain.
4, the detection of Soybean transgenic plant
Excessive or interference GmSWEET6 hypocotyls injection converting material detection：Clip part single plant hypocotyl wound director
The adventitious root gone out extracts RNA and is reversed to cDNA as template, and Plasmid DNA is as positive control, the root system of Non-transgenic soybean
CDNA is as negative control, with upstream specific primer 5 '-CAACCACTACATCGAGACAAGCA-3 ' (SEQ ID NO:17) and
5 '-TCATCAGATCTCGGTGACGGG-3 ' of downstream special primer (SEQ ID NO:18) herbicide resistance gene (Bar bases are expanded
Cause) segment progress PCR detections.In addition, further with quantitative PCR detection is excessive or the effect of interference, with big in quantitative PCR assays
Beans house-keeping gene EF-1a as described above is reference gene (primer sequence：SEQ ID NO:1,2), relative expression quantity is purpose base
Because of GmSWEET6 (primer sequences：SEQ ID NO:3,4) ratio of expression quantity and house-keeping gene expression quantity.It is detected by PCR
And quantitative PCR confirms to obtain effective different transgenic lines.As a result see Fig. 4.
5, the influence of excess or interference GmSWEET6 to Soybean transgenic plant phosphorus content：Harvest Arbuscular Mycorrhizal Fungi
45 days different transgenic lines simultaneously measure phosphorus content, including：Overground part phosphorus content and root phosphorus content etc..
Fig. 5 a, b are excessive or influences of the GmSWEET6 to Soybean transgenic plant phosphorus content is expressed in interference.Wherein plant is adopted
It is trained and is tested with sand, in low-phosphorous (50 μM of KH2PO4) under conditions of, after not being inoculated with (NM) or being inoculated with clump branch bacterium (Ri) processing 45 days
Harvest, the K of low-phosphorous processing+Use K2SO4Polishing.As a result it shows：In the case where being inoculated with Ri, compared with the control, overexpression
After GmSWEET6, root phosphorus content is remarkably decreased, and after interfering GmSWEET6, root phosphorus content significantly rises.
6, the influence of excess or interference expression GmSWEET6 gene pairs Soybean transgenic plant biomass：
Biomass measures：1 percent balances weigh overground part and root samples fresh weight, all samples are dried at 105 DEG C
Case water-removing is placed on 75 DEG C in 30 minutes, and drying to constant weight, weighs dry weight.
Fig. 5 c, d are excessive or influences of the GmSWEET6 to Soybean transgenic plant biomass is expressed in interference.Wherein plant is adopted
It is trained and is tested with sand, in low-phosphorous (50 μM of KH2PO4) under conditions of, after not being inoculated with (NM) or being inoculated with clump branch bacterium (Ri) processing 45 days
Harvest, the K of low-phosphorous processing+Use K2SO4Polishing.As a result it shows：In the case where being inoculated with Ri, compared with the control, overexpression
After GmSWEET6, root biomass is remarkably decreased, and after interfering GmSWEET6, root biomass significantly rises.Show in plant
With the carbon distribution of Mycorrhizal Symbiosiss process, GmSWEET6 may play vital effect, may be planted by regulating and controlling host
Carbon input of the object into Mycorrhizal Symbiosiss system, influences plant and is absorbed to the mycorhiza approach of phosphorus, to influence host plant growth.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, it is other it is any without departing from the spirit and principles of the present invention made by changes, modifications, substitutions, combinations, simplifications,
Equivalent substitute mode is should be, is included within the scope of the present invention.
Sequence table
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<120>The application of soybean sucrose transporter important gene GmSWEET6
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Claims (7)
1. applications of the soybean sucrose transporter important gene GmSWEET6 in terms of Mycorrhizal Symbiosiss.
2. application according to claim 1, it is characterised in that：Soybean sucrose transporter important gene GmSWEET6 is planting
Application in object breeding.
3. application according to claim 1, it is characterised in that：Soybean sucrose transporter important gene GmSWEET6 is being trained
Educate the application in genetically modified plants.
4. application according to claim 1, it is characterised in that：Soybean sucrose transporter important gene GmSWEET6 is being adjusted
Control the application during plant is formed with mycorrhizal fungi beneficial to symbiosis.
5. application according to claim 1, it is characterised in that：Soybean sucrose transporter important gene GmSWEET6 is making
Standby regulation and control plant and the application in mycorrhizal fungi symbiosis preparation.
6. according to claim 2~4 any one of them application, it is characterised in that：
The plant is dicotyledonous legume.
7. application according to claim 6, it is characterised in that：
The dicotyledonous legume is soybean.
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| CN110144358A (en) * | 2019-05-28 | 2019-08-20 | 华中农业大学 | A kind of clover SWEET1b gene and its application in promotion clover root system mycorhiza rate |
| CN110669782A (en) * | 2019-10-10 | 2020-01-10 | 南京农业大学 | Application of soybean sugar transporter gene GmSWEET39 |
| CN111154772A (en) * | 2020-02-09 | 2020-05-15 | 南京农业大学 | Pear sugar transport gene PbSWEET4 and application thereof |
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| CN110669782A (en) * | 2019-10-10 | 2020-01-10 | 南京农业大学 | Application of soybean sugar transporter gene GmSWEET39 |
| CN110669782B (en) * | 2019-10-10 | 2022-11-01 | 南京农业大学 | Application of soybean sugar transporter gene GmSWEET39 |
| CN111154772A (en) * | 2020-02-09 | 2020-05-15 | 南京农业大学 | Pear sugar transport gene PbSWEET4 and application thereof |
| CN111154772B (en) * | 2020-02-09 | 2022-10-04 | 南京农业大学 | Pear sugar transport gene PbSWEET4 and application thereof |
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