CN106397557A - Sweet potato adversity-tolerant associated protein IbFBA2, coding gene, recombinant vector and application - Google Patents
Sweet potato adversity-tolerant associated protein IbFBA2, coding gene, recombinant vector and application Download PDFInfo
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Abstract
The invention discloses a sweet potato adversity-tolerant associated protein IbFBA2, a coding gene, a recombinant vector and application. The IbFBA2 protein is a protein formed by an amino acid sequence expressed as a sequence 2; the coding gene is a DNA molecule expressed by a sequence 1; the recombinant vector containing the coding gene is a recombinant vector obtained by inserting DNA molecules expressed by 28-1584 nucleotides of 5' terminal of the sequence 1 between two attR loci of pWGB12. The IbFBA2 protein and the coding gene and the recombinant vector thereof can regulate plants to respond to ABA stress; a transgenosis plant capable of responding to ABA treatment can be obtained by introducing the DNA molecule coding the IbFBA2 protein into a target plant. The IbFBA2 protein and the coding gene thereof have important application value in improvement of response to adversity stress of the plants; the foundation is established for cultivation of adversity-tolerant transgenosis plants.
Description
Technical field
The invention belongs to genetic engineering field and in particular to sweet potato conversion-resisting resisting related protein IbFBA2, encoding gene, restructuring carry
Body and application.
Background technology
In agricultural production, stable high yield is the effective way of Ensuring Food Safety.Day with Global Ecological such as weathers
Benefit deteriorates, and the extreme environment such as arid, salt marsh usually causes extensive damage to the yield and quality of crops.Abscisic acid
(absicsic acid, ABA) is a kind of important plant hormone, in the eubolism and growth and development process of plant
Participate in multi-signal regulatory pathway.Abscisic acid is also referred to as " adversity plant hormone ", (includes dry resisting external environment stress
The biotic such as the abiotic stress such as drought, low temperature, salt marsh, infiltration and pest and disease damage) during play vital effect.Cause
This, the molecular mechanism that understanding plant responding ABA is processed, and then the resistance of reverse of plant is regulated and controled by the method for genetic improvement, become
The emphasis studied for current molecular biology and Agricultural Scientist and focus.
By Protocols in Molecular Biology, excavate plant responding ABA and process correlation function gene, inverse stable yields crop resistance to cultivation
New varieties have important using value.
Content of the invention
First purpose of the present invention is to provide and sweet potato conversion-resisting resisting related protein IbFBA2.
For achieving the above object, IbFBA2 albumen provided by the present invention, from sweet potato [Ipomoea batatas
(L.) Lam], for the protein of following (1) or (2)：
(1)SEQ ID NO：The protein of the amino acid sequence composition shown in 2；
(2)SEQ ID NO：Amino acid sequence shown in 2 is through the replacement of one or several amino acid residues and/or disappearance
And/or add and by (1) derived from the protein related to plant stress tolerance.
SEQ ID NO in sequence table：Amino acid sequence shown in 2 is made up of 398 amino acid residues.
In above-mentioned protein, the replacement of one or several amino acid residue and/or disappearance and/or interpolation refer to few
Replacement and/or disappearance and/or interpolation in ten amino acid residues.
Second object of the present invention is to provide the gene encoding described IbFBA2 albumen.
For achieving the above object, the encoding gene of IbFBA2 albumen provided by the present invention, is any in following (1)-(4)
A kind of DNA molecular：
(1)SEQ ID NO：1 from 5 ' ends the DNA molecular shown in the 97th to the 1293rd nucleotides；
(2)SEQ ID NO：DNA molecular shown in 1；
(3) the DNA molecular hybridization limiting with (1) or (2) under strict conditions and coding and plant stress tolerance correlative protein
DNA molecular；
(4) DNA sequence dna being limited to (1) or (2) is had more than 90% homology and encodes and the related egg of plant stress tolerance
White DNA molecular.
Described stringent condition is with 6 × SSC, the solution of 0.5%SDS, hybridizes at 65 DEG C, then with 2 × SSC, 0.1%
SDS and 1 × SSC, 0.1%SDS respectively wash film once.
Wherein, sequence SEQ ID NO：1 length is 1617bp, and its exploitation reading frame (ORF) is from 5 ' ends the 97th
To the 1293rd bit base, its encoding amino acid sequence is SEQ ID NO in sequence table for position：Albumen shown in 2.
Third object of the present invention is to provide the recombinant vector of the encoding gene containing described IbFBA2 albumen.
The recombinant vector that the present invention provides is by SEQ ID NO：1 from shown in 5 ' the 28th to the 1584th nucleotides in end
DNA molecular is inserted into the recombinant vector obtain between two attR sites of pGWB12.
Fourth object of the present invention is to provide described IbFBA2 albumen, encoding gene and the weight containing described encoding gene
Application in regulation and control plant stress tolerance for the group carrier.
In above-mentioned application, the patience that described resistance of reverse is coerced for ABA；Described plant is dicotyledon, described dicotyledonous
Plant can be arabidopsis (Arabidopsis thaliana)；Described regulation and control plant stress tolerance is regulation and control plant responding ABA stress,
Described regulation and control plant responding ABA coerces the growing way for controlling plant root on the 1/2MS culture medium containing 20mmol ABA；Described
Growing way be that the quantity of the statistics length of plant roots and root embodies.
5th purpose of the present invention is to provide described IbFBA2 albumen, encoding gene and the weight containing described encoding gene
Application in cultivating genetically modified plants for the group carrier.Specifically, described genetically modified plants have following proterties：The length of root is more right
Relatively compare more according to longer, root quantity；Described plant is arabidopsis.
6th purpose of the present invention is to provide a kind of method cultivating the genetically modified plants that response ABA is processed.
The method cultivating genetically modified plants provided by the present invention, is to lead the DNA molecular encoding above-mentioned IbFBA2 albumen
Enter purpose plant, obtain genetically modified plants, described genetically modified plants are than purpose plant more resistant to environment stress.
In said method, the described DNA molecular encoding above-mentioned IbFBA2 albumen is to import mesh by above-mentioned recombinant vector
Plant in.
In said method, described purpose plant is wildtype Arabidopsis thaliana (Col_0).
The invention provides a kind of IbFBA2 albumen and its encoding gene, by this channel genes wildtype Arabidopsis thaliana, obtain
To turning IbFBA2 Arabidopsis plant, result proves that IbFBA2 can promote purpose plant responding ABA process, increases resistance of reverse,
The length of root is considerably longer than and compares, and the quantity of root is significantly more than and compares.Illustrate that IbFBA2 albumen and its encoding gene ring in plant
Answer in ABA processing procedure and play an important role.The IbFBA2 albumen that the present invention provides and its encoding gene are resistance in raising plant
Inverse property aspect has important theory significance and using value.
Brief description
Fig. 1 is the PCR testing result figure turning IbFBA2 Arabidopsis plant；
Fig. 2 is to turn IbFBA2 arabidopsis to respond the phenotypic results figure that ABA is processed；
Fig. 3 is to turn IbFBA2 gene arabidopsis long comparison diagram of root under different ABA concentration；
Fig. 4 is to turn IbFBA2 gene arabidopsis radical amount comparison diagram under different ABA concentration.
Specific embodiment
With instantiation, the present invention is described in further detail below in conjunction with the accompanying drawings.
In following examples, experimental technique used if no special instructions, is conventional method.
Material used, reagent etc. in following examples, if no special instructions, all commercially obtain.
Embodiment 1 responds, with sweet potato, the acquisition that ABA processes GAP-associated protein GAP IbFBA2 and its encoding gene
Experiment material：By sweet potato variety Xu's 781 potato wedgess (being provided by Xuzhou Sweetpotato Center) in seedbed vernalization, treat
Growth takes the fresh blade of 1-2 piece, liquid nitrogen flash freezer, -80 DEG C of preservations after about 30 days.
1st, blade Total RNAs extraction and purifying
RNA extract used by solution, vessel etc. all through the specially treated of DEPC water, its operation in strict accordance with《Molecular cloning
Experiment guide (second edition)》The requirement of (2005-3-1, J. Pehanorm Brooker etc., Science Press) relevant RNA operation is carried out, sternly
Anti- RNase pollution.
Total RNA extraction reagent box (the animal total serum IgE rapid extraction kit (centrifugal column being produced using Shanghai JaRa company
Type), catalogue is GK3016) carry out the extraction of RNA, comprise the following steps that：
1) by sample grind into powder in liquid nitrogen, in the centrifuge tube of transferase 45 0.0mg to 1.5ml RNase-Free, and
Plus 1.0ml RnaEX (Trizol)；
2) vortex oscillation 30sec, room temperature stands 5min, with abundant lysate sample；
3) 200.0 μ l chloroforms, vortex oscillation 30sec are added in ventilating kitchen, room temperature stands 5min；
4) on desk centrifuge, 12000rpm, 4 DEG C of centrifugation 10min, so that solution is fully layered；
5) directly draw the supernatant of about 500.0 μ l to centrifugal column, add the absolute ethyl alcohol of 200.0 μ l, and fully mix,
Room temperature standing 2min (might have flocculent deposit to produce)；
6), under normal temperature condition, 8000rpm is centrifuged 1min；
7) carefully take out pillar, outwell filtrate, and pillar is put back in collection post, add 600.0 μ l buffer RWA,
8000rpm, room temperature is centrifuged 30sec, outwells filtrate；
8) step repeating 7；
9) outwell filtrate, pillar is put back to collecting pipe, 12000rpm is centrifuged 1min；
10) carefully take out pillar, put in the 1.5ml centrifuge tube of new RNase-Free, add 50.0 μ in post film central authorities
The DEPC-H2O of l, places 2min at 60 DEG C；
11) 12000rpm centrifugation 1min, the solution in collecting pipe is RNA sample, can immediately using or -80 DEG C of preservations.
Take 6 μ l RNA for electrophoresis detection, take 2 μ l RNA for concentration mensuration, remaining RNA saves backup in -80 DEG C.
Quality (OD with UV spectrophotometer measuring RNA260) and purity (OD260/OD280)；And use 1.2% Denaturing Agarose Gel
Electrophoresis detection RNA integrality.
2nd, the synthesis of cDNA first chain
The total serum IgE that will extract, is become by M-MLV reverse transcriptase (Promega, catalog number is M1701) reverse transcription
First-strand cDNA, comprises the following steps that：
1) take total serum IgE 2.0 μ g in PCR pipe, add the Oligo (dT) 18 (0.3 μ g/ μ l) of 2.9 μ l, use RNase-
Free H2O complements to 15.0 μ l, mixes centrifuge tube, and of short duration centrifugation；
2) utilize PCR instrument temperature control, heat 70 DEG C of centrifuge tube, 5min, be immediately placed in cooled on ice 2min；
3) it is subsequently adding following reagent：5.0 μ l M-MLV 5 × Reaction Buffer, 1.2 μ l dNTP mix
(10mM), 0.6 μ l RNase Inhibitor (40U/ μ l), 1.1 μ l M-MLV reverse transcriptase (200.0U/ μ l), plus RNase-
Free H2It is 25.0 μ l that O is settled to cumulative volume；
4) gentle centrifugation after mixing, 42 DEG C of incubation 60min in PCR instrument, you can obtain First-strand cDNA；
5) take in 4.0 μ l to 0.2ml centrifuge tubes, and dilute 10 times for subsequent experimental or -20 DEG C of preservations of placement.
3rd, the cDNA coded sequence clone of IbFBA2 albumen
According to the result of sweet potato transcript profile sequencing, the cDNA sequence finding IbFBA2 albumen is compared by homologous sequence, and
Design forward primer 1 and reverse primer 2 carry out IbFBA2 albumen cDNA coded sequence clone.Primer sequence is as follows：
Primer 1：5’–GAGGAGATAAGATCGAATAGTAAAA-3’(SEQ ID NO：3)
Primer 2：5’–GGAATACAAGATAAACCATAACTGA-3’(SEQ ID NO：4)
The ready cDNA with step 2, as template, enters performing PCR reaction with forward primer 1 and reverse primer 2.
PCR reaction system is as follows：10.0 μ l 5 × PrimeSTAR GXLbuffer, 4.0 μ l dNTPs (2.5mM), 1.0 μ
L PrimeSTAR GXL archaeal dna polymerase (1.25U/ μ l) (purchased from Dalian TAKARA company, catalogue is R050Q), 100ng
CDNA, the forward and reverse primer of 1.0 μ l (10.0mM), ddH2O is supplemented to final volume 50.0 μ l.Thermal circulation parameters are：94℃3min；
94 DEG C of 30sec, 57 DEG C of 30sec, 72 DEG C of 1min30sec, 35 circulations；72 DEG C of 10min, 1 circulation；4 DEG C preservation, reaction be
Complete in Eppendorf (Mastercycler Nexus) PCR instrument.Amplified production 1.2% agarose on Horizontal electrophoresis tank coagulates
Gel electrophoresis detect, obtain the amplified fragments of 1557bp length.
Through sequencing, this PCR primer has SEQ ID NO：Nucleotides shown in 1, the coding of the gene shown in this sequence
Area is SEQ ID NO in sequence table：1 from 5 ' the 97th 1293 nucleotides in end；SEQ ID NO in sequence table：1 by 1617 alkali
Base forms；The albumen of this gene code is named as IbFBA2, and the amino acid sequence of this albumen is the SEQ ID NO in sequence table：
2；SEQ ID NO in sequence table：2 are made up of 398 amino acid residues.
Application in improving plant ABA stress tolerance for the embodiment 2IbFBA2 albumen
1st, the structure of plant expression vector
Coded sequence according to sweet potato IbFBA2 albumen cDNA and the Gateway pCR8/GW/TOPO TA of carrier construction
Cloning kit (Invitrogen, catalog number is 45-0642) technology, the PCR primer that embodiment 1 is obtained carries out pure
Change, and mix with pCR8/GW/TOPO carrier, proceed to competent escherichia coli cell Trans-T1 (biological purchased from the full formula in Beijing gold
Technology Co., Ltd., catalog number is CD501) in, it is coated onto the solid LB plating medium containing spectinomycin (Spe) resistance
On, under the conditions of 37 DEG C, overnight incubation treats that bacterium colony grows, and selects that monoclonal makees bacterium colony forward primer 1 and reverse primer 2 enters performing PCR
Reaction.PCR reaction system is as follows：10.0 μ l health are 2 × Taq mix (is ShiJi Co., Ltd purchased from Beijing health, catalog number is
CW0716A), 1.0 μ l monoclonal diluting colonies liquid, 1.0 μ l forward primers (SEQ ID NO in sequence table：3), reverse primer (sequence
SEQ ID NO in list：4) (10.0mM), ddH2O is supplemented to final volume 20.0 μ l.Thermal circulation parameters are：94℃3min；94℃
30sec, 55 DEG C of 30sec, 72 DEG C of 1min, 35 circulations；72 DEG C of 10min, 1 circulation；4 DEG C of preservations, reaction is in Eppendorf
Complete in (Mastercycler Nexus) PCR instrument.Amplified production 1.2% agarose gel electrophoresis inspection on Horizontal electrophoresis tank
Survey.Select that PCR detection has a band and the very bright corresponding monoclonal of band serves sea raw work sequencing, determine the direction of fragment insertion with
And the sequence of fragment no makes a variation.
By being sequenced, errorless carrier is named as pCR8/GW/TOPO::IbFBA2, by carrier pCR8/GW/TOPO::IbFBA2
Carry out LR reaction with plant expression vector pGWB12 (being provided by sweet potato research institute of the Chinese Academy of Agricultural Sciences), obtain recombinant vector
pGWB12:IbFBA2, as plant expression vector.
By above-mentioned recombinant vector pGWB12:IbFBA2 conversion E. coli competent Trans-T1 is (purchased from the full formula in Beijing gold
Bioisystech Co., Ltd, catalog number is CD501), it is coated onto the culture of the solid LB flat board containing kanamycins (Kan) resistance
On base, 37 DEG C of culture 16h, to recombinant vector pGWB12:IbFBA2 enters performing PCR amplification to be identified and carries out sequence verification.Sequencing knot
Fruit shows, recombinant vector pGWB12:IbFBA2 is by SEQ ID NO in sequence table：1 holds the 28th to shown in the 1584th from 5 '
The carrier obtaining between two attR sites of nucleotides inserted pGWB12.
2nd, plant expression vector conversion Agrobacterium
1) (stepping its biotechnology purchased from Shanghai has to take out the GV3101 competent cell of 100 μ l from -80 DEG C of low temperature refrigerators
Limit company, catalog number is CH5002A), it is placed in thawed on ice, add the plant expression vector that 1.0 μ g above-mentioned steps 1 obtain
pGWB12:IbFBA2, mixes；
2) liquid nitrogen frozen 5min, 37 DEG C of incubation 5min；
3) the LB fluid nutrient medium of 250.0 μ l, 28 DEG C of culture 2h are added；
4) 200.0 μ l bacterium solution are taken in containing antibiotic (100.0 μ g/ml rifampin (Rif), 50.0 μ g/ml kanamycins
(Kan) it is coated with LB solid medium) uniformly, cultivate 1-2 days in 28 DEG C；
5) picking single bacterium colony performing PCR identification of going forward side by side (identifies that primer is primer 1 (SEQ ID NO in sequence table：3) and primer 2
(SEQ ID NO in sequence table：4) bacterium colony), obtaining 1557bp fragment is positive bacterium colony；
6) positive single bacterium colony is inoculated in the LB fluid nutrient medium containing 100.0 μ g/ml Rif and 50 μ g/ml Kan, 28
DEG C, the shaking table culture of 150rpm reach 0.8 to OD value, that is, obtained can be used for the agrobacterium liquid of subsequent transformation work.
3rd, turn the acquisition of IbFBA2 arabidopsis
The coded sequence of IbFBA2cDNA is imported to wild type with agriculture bacillus mediated method and intends in southern plant by conversion, tool
Body method is as follows：
1) by after ready agrobacterium liquid centrifugation in step 2, outwell supernatant, add same volume contain 5.0% sucrose,
The aaerosol solution of 0.02%silwet L-77 surfactant；
2) draw above-mentioned agrobacterium suspension with liquid-transfering gun, drip to and (remove open on the bud of arabidopsis and axillary bud
Flower)；
3) plant handled well is placed in the plastic tub of printing opacity, inwall spray a certain amount of water, and cover preservative film with
Maintain higher levels of humidity, 24h is cultivated in normal illumination；
4) next day removes preservative film, and after normally cultivating 1 month, individual plant harvests and obtains T1In generation, turns IbFBA2 arabidopsis seed；
5) to T1Carry out disinfection for seed, be seeded into hygromycin containing 40mg/L with Top agar (0.1%agar)
(Hyg) screened on 1/2MS culture medium；
6) sprouting situation according to root, the individual plant selecting long root is transplanted in matrix, and after normally cultivating 1 month, PCR checks
Choose transgenic positive plant and individual plant harvests, obtain T2In generation, turns IbFBA2 arabidopsis seed；
4th, turn the ABA tolerance identification of IbFBA2 arabidopsis
Numbering is to sow after T11, T48, T50 transgenic arabidopsis seed and wildtype Arabidopsis thaliana (WT) seed disinfection
To on 1/2MS culture medium, light culture 48-72 hour under the conditions of 4 DEG C, it is placed in culture in illumination box after taking-up, treat two panels
After cotyledon opens completely, select the uniform seedling cultivation of growing way to the square culture of 1/2MS containing 0mM (comparison) and 20mM ABA
On base (100mm × 100mm), vertically cultivate in illumination box, observe arabidopsis growing state.Each strain plants 3 seedlings,
Two process each three biology and repeat, and statistics is averaged.
Extract wildtype Arabidopsis thaliana and T with Edwards Solution respectively2In 3 plants of generation, turns the genome of IbFBA2 arabidopsis
DNA, as template, enters performing PCR detection, and IbFBA2 gene primer used is：Primer 1 (SEQ ID NO：3) and primer 2 (SEQ
ID NO：4) AtTub gene primer, used is：Primer 3：5’-AGCAAATGTGGGACTCCAAG-3’(SEQ ID NO：5)
With primer 4：5’-CACCTTCTTCATCCGCAGTT-3’(SEQ ID NO：6).PCR reaction system is as follows：10.0μl2×Taq
Mix (being ShiJi Co., Ltd purchased from Beijing health, catalog number is CW0716A), 1.0 μ l DNA (50ng/ μ l), 1.0 μ l forward directions are drawn
Thing (SEQ ID NO in sequence table：3), reverse primer (SEQ ID NO in sequence table：4) (10.0mM), ddH2O is supplemented to whole body
Long-pending 20.0 μ l.Thermal circulation parameters are：94℃3min；94 DEG C of 30sec, 55 DEG C of 30sec, 72 DEG C of 1min, 35 circulations；72℃
10min, 1 circulation；4 DEG C of preservations.
See that (swimming lane M is Marker to Fig. 1 from electrophoresis detection amplification；Swimming lane P is positive control (recombinant plasmid
pGWB12::IbFBA2)；W is blank；WT is negative control (Col_0 arabidopsis)；Swimming lane T11, T48 and T50 are to turn
IbFBA2 gene arabidopsis) as can be seen that obtain 1557bp for positive T2In generation, turns IbFBA2 arabidopsis, shows IbFBA2 gene
It has been incorporated in arabidopsis gene group, and proved that these regeneration plants are positive transgenic plant.
Result is as shown in Figures 2 and 3 it can be seen that numbering on the culture medium of 20mM ABA is turning of T11, T48, T50
The length of gene arabidopsis root long significant elongation than the root of adjoining tree, shows that its resistance of reverse is remarkably reinforced.As can be seen from Figure 4
On the culture medium of 20mM ABA, numbering be the transgenic arabidopsis root of T11, T48, T50 number relatively compare more.
Above-mentioned test result indicate that, IbFBA2 albumen and its encoding gene can strengthen the patience of plant responding ABA stress,
Strengthen plant stress tolerance.
Claims (9)
1. a kind of IbFBA2 albumen is it is characterised in that this albumen source is in sweet potato, for the protein of following (1) or (2)：
(1)SEQ ID NO：The protein of the amino acid sequence composition shown in 2；
(2)SEQ ID NO：Amino acid sequence shown in 2 through one or several amino acid residues replacement and/or disappearance and/
Or add and by (1) derived from the protein related to plant stress tolerance.
2. the encoding gene of the IbFBA2 albumen described in claim 1.
3. the encoding gene described in claim 2 is it is characterised in that described gene is that any one of (1)-(4) DNA divides as follows
Son：
(1)SEQ ID NO：1 from 5 ' ends the DNA molecular shown in the 97th to the 1293rd nucleotides；
(2)SEQ ID NO：DNA molecular shown in 1；
(3) the DNA molecular hybridization limiting with (1) or (2) under strict conditions and the DNA of coding and plant stress tolerance correlative protein
Molecule；
(4) DNA sequence dna being limited with (1) or (2) is had more than 90% homology and encodes and plant stress tolerance correlative protein
DNA molecular.
4. the recombinant vector containing encoding gene described in Claims 2 or 3.
5. recombinant vector according to claim 4 is it is characterised in that described recombinant vector is by SEQ ID NO：1 from 5 '
The DNA molecular shown in the 28th to the 1584th nucleotides is held to be inserted into the restructuring obtain between two attR sites of pGWB12
Carrier.
6. described in the IbFBA2 albumen described in claim 1 and the encoding gene described in Claims 2 or 3 and claim 4 or 5
Recombinant vector regulation and control plant stress tolerance in application；The patience that described resistance of reverse is coerced for ABA；Described plant is dicotyledonous
Plant, described dicotyledon is arabidopsis；Described regulation and control plant stress tolerance is regulation and control plant responding ABA stress, described regulation and control
Plant responding ABA coerces the growing way for controlling plant root on the 1/2MS culture medium containing 20mmol ABA；The growing way of described is
The quantity of the length of statistics plant roots and root embodies.
7. described in the IbFBA2 albumen described in claim 1 and the encoding gene described in Claims 2 or 3 and claim 4 or 5
Recombinant vector cultivate response ABA process genetically modified plants in application；Described plant is arabidopsis.
8. a kind of method cultivating genetically modified plants is it is characterised in that the method is by albumen described in coding claim 1
DNA molecular imports purpose plant, obtains genetically modified plants；Described genetically modified plants are than described purpose Plant Tolerance environment stress；
Described purpose plant is col_0 arabidopsis.
9. cultivation genetically modified plants method according to claim 8 is it is characterised in that described in described coding claim 1
The DNA molecular of albumen is to be imported in described purpose plant by the recombinant vector described in claim 4 or 5.
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Cited By (1)
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| CN112725367A (en) * | 2021-03-01 | 2021-04-30 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | Sweet potato sucrose invertase gene IbINV and application thereof |
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| CN112725367A (en) * | 2021-03-01 | 2021-04-30 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | Sweet potato sucrose invertase gene IbINV and application thereof |
| CN112725367B (en) * | 2021-03-01 | 2022-07-15 | 江苏徐淮地区徐州农业科学研究所(江苏徐州甘薯研究中心) | Sweet potato sucrose invertase gene IbINV and application thereof |
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