CN106520799A - Non-specific phospholipase C gene GhNPC1b of upland cotton and applications of non-specific phospholipase C gene GhNPC1b - Google Patents
Non-specific phospholipase C gene GhNPC1b of upland cotton and applications of non-specific phospholipase C gene GhNPC1b Download PDFInfo
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Abstract
The invention discloses a non-specific phospholipase C gene GhNPC1b of upland cotton. The cDNA nucleotide sequence of the gene is shown as SEQ ID No.1, and the amino acid sequence encoded by the gene is shown as SEQ ID No.2. The invention further discloses the applications of the gene and a plant expression vector pCAMBIA1300-GhNPC1b-EPSP containing the gene in improving the stress resistance of cotton. The experiment proves that the expression of the gene provided by the invention is induced by the stress of low phosphorous content, salt and drought, and the genetically modified cotton experiment shows that the adverse-resistant characteristic of the genetically modified cotton can be obviously improved, which indicates that the GhNPC1b gene has the great potential when used for culturing adverse-resistant genetically modified crops.
Description
Technical field
The present invention relates to a kind of Gossypium hirsutum L. non-specificity phospholipase C gene GhNPC1b and its improve Cotton Gossypii resistance in
Application.Belong to plant genetic engineering field.
Background technology
Cotton Gossypii is most important natural fiber in the world, and the abiotic stress such as arid, saline and alkaline affects the growth of Cotton Gossypii, finally
Output of cotton is caused to reduce.In the case where cultivated area only subtracts and do not increase, some places occur in that grain and cotton strives the phenomenon on ground.In people
The long korneforos of class development of civilization, wears the clothes and has a meal and be of equal importance.It is that cultivation resistance is stronger that solution grain and cotton strives the best approach on ground
Elite cotton kind, and Efficient Development utilize arid biogeographic zone, salt-soda soil or soil leanness soil.
Non-specific phospholipase C, mainly with phosphatidylcholine as substrate, generates diacylglycerol and phosphocholine.Arabidopsiss
In, there are 6 members (Nakamura et al.2005) in non-specific phospholipase C gene family.So far, there is in succession text within 2005
Offer the signal transduction and degeneration-resistant process (Nakamura et of report plant non-specificity phospholipase C involved in plant hormone
al.2005；Peters et al.2010；Wimalasekera et al.2010；Kocourkova et al.2011；
Pokotylo et al.2013；Nakamura 2014；Peters et al.2014；et al.2015；Pejchar
et al.2015；Hong et al.2016).However, object of study is model organism arabidopsiss mostly, non-spy in other species
The functional study of different in nature phospholipase C is very few.In order to be best understood from the function of plant non-specificity phospholipase C, to the non-spy of Cotton Gossypii
Different in nature phospholipase C gene family carries out research and is very important, but retrieves discovery so far, and no any Cotton Gossypii is non-specific
Property phospholipase C related report.
The content of the invention
For the deficiencies in the prior art, it is an object of the invention to provide a kind of Gossypium hirsutum L. non-specificity phospholipase C gene
GhNPC1b and its application in Cotton Gossypii resistance is improved.
Gossypium hirsutum L. non-specificity phospholipase C gene of the present invention, it is characterised in that：The unnamed gene is that Gossypium hirsutum L. is non-
Specific phospholipase C gene GhNPC1b, the cDNA nucleotide sequences of the gene as shown in SEQ ID No.1, its coding amino
Acid sequence is as shown in SEQ ID No.2.
The present invention clones Gossypium hirsutum L. non-specificity phospholipase C gene GhNPC1b first, and experimental analysiss confirm：GhNPC1b
On A03 chromosomes, the number of aminoacid is 486, containing 3 exons, 2 introns for gene mapping.MEME analyses find,
GhNPC1b has 17 motif.Motif to obtaining further goes to analyze in ExPASy, and it is phosphoric acid that discovery has 6 motif annotations
Ester structure domain.With the conserved domain of NCBI Conserved Domain search and SMART on-line analyses GhNPC1b, send out
Existing GhNPC1b contains phosphate ester structure domain.Promoter cis acting element analysis is carried out to GhNPC1b, finds opening for GhNPC1b
Containing the cis acting element that or hormone related to stress is related in mover.Including MBS (drought stress is related), HSE (heat stresses
It is related), LTR (low temperature stress is related), Box-W1 (fungus induction is related) ABRE and CE3 (abscisic acid is related), GARE-motif
(gibberellins are related), TCA-element (salicylic acid is related), TGACG-motif and CGTCA-motif (methyl jasmonate phases
Close).
The expression pattern analysis of Different Organs are carried out to GhNPC1b, and real-time fluorescence quantitative PCR result shows that GhNPC1b is in leaf
Expression in piece is of a relatively high.Carry out low-phosphorous (5 μM), salt (200mM NaCl), arid (20% to four leaf stage Cotton Gossypii respectively
PEG6000) and ABA (200 μM) Stress treatment, respectively process 0,1,2,3,6,9 and 12h when drawn materials, position of drawing materials
For root.Extract RNA and be reversed to cDNA, real-time fluorescence quantitative PCR result shows, under low-phosphorus stress and salt stress, GhNPC1b
Expression is raised, wherein, during low-phosphorus stress 6h, the phenomenon for showing to lower of GhNPC1b；Drought stress process 1h and
During 2h, GhNPC1b expressions are raised, and show the phenomenon of slight downward after 2h；Under ABA Stress treatments, GhNPC1b expression water
It is flat to lower.The expression of GhNPC1b genes is by low-phosphorous, salt and the stress-inducing of arid.
Present invention also offers a kind of plant expression containing above-mentioned Gossypium hirsutum L. non-specificity phospholipase C gene GhNPC1b
Carrier pCAMBIA1300-GhNPC1b-EPSP.
Applications of the Gossypium hirsutum L. non-specificity phospholipase C gene GhNPC1b of the present invention in Cotton Gossypii resistance is improved.
Plant expression vector pCAMBIA1300-GhNPC1b-EPSP of the present invention answering in Cotton Gossypii resistance is improved
With.
Wherein：The resistance refers to Salt And Alkali Tolerance, drought resisting and/or Tolerant to low P.The cotton variety is spring cotton, summer cotton, often
Rule cotton or hybrid cotton.
When GhNPC1b overexpression structures are built, the cDNA nucleotide sequences of GhNPC1b are inserted into into plant expression vector
In pCAMBIA1300-EPSP, plasmid pCAMBIA1300-GhNPC1b-EPSP is obtained.After enzyme action identification is correct, plasmid is imported
In agrobacterium tumefaciens AGL1 or LBA4404, for Genetic Transformation in Higher Plants.
By agriculture bacillus mediated transgenic method, target gene is turned by receptor of the shoot apical meristem cell of Cotton Gossypii
Enter in cotton cells, that is, obtain transfer-gen plant.Determined by Molecular Detection and plant resistance and select the stable table of genes of interest
Up to and the transfer-gen plant that significantly improves of plant resistance.By pot experiment and land for growing field crops resistance determination test, you can filter out
The transgenic breeding material that resistance is significantly improved.The elite clone of acquisition is expected to be used for cotton breeding or production practices.
The invention has the beneficial effects as follows：
The present invention clones non-specific phospholipase C gene GhNPC1b first from Gossypium hirsutum L..The expression of the gene is by low
The induction of phosphorus, salt and drought stress.The present invention successfully constructs pCAMBIA1300-GhNPC1b-EPSP plant expression vectors simultaneously,
The plant expression vector is transformed in Cotton Gossypii and is confirmed, gene of the present invention can significantly improve the degeneration-resistant spy of transgene cotton
Property.Indication, GhNPC1b genes are applied to cultivation resistant transgenic crops and have great potentiality.
Description of the drawings
Fig. 1：The structural representation of plasmid pCAMBIA1300-GhNPC1b-EPSP.
Fig. 2：GhNPC1b gene organizations specificity analyses
Wherein：TR, main root；LR, lateral root；SM, stem；CL, cotyledon；SL, ageing leaves；EL, young leaflet tablet；ST, stem apex；
BR, bract；PE, petal；OV, ovule.
Fig. 3：Expression pattern analysis of the GhNPC1b genes under abiotic stress.
Specific embodiment
Embodiments of the invention described below.It should be noted that, embodiments of the invention only play illustration to the present invention
Without restriction effect.If no special instructions, it is conventional method.Test material used in following embodiments, such as without special
Illustrate, be routine biochemistry reagent.Quantitative test in following examples, is respectively provided with three repetitions and tests, results averaged.
1. high-fidelity PCR amplification GhNPC1b genes of interest of embodiment
The tender blade of 9807 four leaf stages children and root in Gossypium hirsutum L. is taken, with reference to plant total RNA extraction reagent box (TIANGEN, north
Capital) description, plant total serum IgE is extracted, with PrimeScript RT Reagent Kit (TaKaRa, Dalian) reverse transcription reagent box
Reverse transcription obtains cDNA.High-fidelity PCR amplification is carried out with special primer.Specific primer sequences are as follows：
GhNPC1b-F：GGTACCATGGAAAACCGTTCTTTTGA
GhNPC1b-R：GAGCTCCTAATAGGCTTCCACATGTT
PCR reacts cumulative volume for 25 μ L, and reaction system is：5 μ L 5 × PS Buffer, 2 μ L dNTP, 0.5 μ L forward directions draw
Thing, 0.5 μ L reverse primers, 0.25 μ L Prime STARase, 1 μ L dilute ten times of cDNA templates, ddH2O polishings to 25 μ L.
PCR amplification conditions, 95 DEG C of 3min of denaturation, 35 circulations：98 DEG C, 10sec；56 DEG C, 5sec；72 DEG C, 90sec,
Last 72 DEG C of extensions 6min.
PCR primer Jing agarose gel DNA QIAquick Gel Extraction Kits recovery (TIANGEN, Beijing) after purification, are connected to
On pEASY-Blunt Cloning vector (TRANSGEN, Beijing).Then convert escherichia coli TransT1 (TRANSGEN,
Beijing), carry out sequencing analysis.By above-mentioned steps, the cDNA of Gossypium hirsutum L. non-specificity phospholipase C gene GhNPC1b is obtained
Nucleotide sequence.
The grand result of GhNPC1b gene cDNA nucleotide sequence lek is as follows：
The cDNA nucleotides sequences that GhNPC1b genes are obtained by high-fidelity enzymatic amplification are classified as 1461bp, encode 486 amino
Acid, as shown in SEQ ID No.1, the aminoacid sequence of the GhNPC1b albumen of its coding is such as the cDNA nucleotide sequences of GhNPC1b
Shown in SEQ ID No.2.
Embodiment 2.GhNPC1b gene biological bioinformatics analysis
Using information such as ExPASy proteomics server database estimation GhNPC1b molecular weight, isoelectric point, IPs；
GhNPC1b gene structures are analyzed using Gene Structure Display Server 2.0；GhNPC1b is analyzed using MEME
Conservative motif, and with the ScanProsite instruments in ExPASy to obtain motif annotate；Using NCBI
The conserved domain of Conserved Domain search and SMART on-line analyses GhNPC1b, is analyzed using Plant CARE
The promoter of GhNPC1b.
Through bioinformatic analysis, GhNPC1b genes are found, is positioned on A03 chromosomes, the number of aminoacid is
486, containing 3 exons, 2 introns.MEME analyses find that GhNPC1b has 17 motif.Motif to obtaining enters one
Step goes to analyze in ExPASy, and it is phosphate ester structure domain that discovery has 6 motif annotations.With NCBI Conserved Domain
The conserved domain of search and SMART on-line analyses GhNPC1b, it is found that GhNPC1b contains phosphate ester structure domain.It is right
GhNPC1b carries out promoter cis acting element analysis, containing or hormone phase related to stress in the promoter of discovery GhNPC1b
The cis acting element of pass.MBS (drought stress is related), HSE (heat stress is related), LTR (low temperature stress is related), Box-W1
(fungus induction is related) ABRE and CE3 (abscisic acid is related), GARE-motif (gibberellins are related), TCA-element (salicylic acid
It is related), TGACG-motif and CGTCA-motif (methyl jasmonate is related).
Embodiment 3.GhNPC1b tissue specificity is analyzed
Gossypium hirsutum L. (kind be in 9807) is planted in greenhouse after seed is sterilized, and growth conditionss are：30 DEG C/25 DEG C, phase
It is 60-70% to humidity, illumination 14h, dark 10h, photon flux density are 800 μm of ol m-2s-1.When growing to four leaf stage, take
, in liquid nitrogen, -80 DEG C preserve for main root, lateral root, stem, stem apex, cotyledon, ageing leaves and young leaflet tablet；Florescence is grown to, is taken away
In liquid nitrogen, -80 DEG C preserve bract, petal and ovule when spending first day.Plant total serum IgE is extracted respectively, and reverse transcription is obtained
CDNA, 10 times of dilution are analyzed for real-time fluorescence quantitative PCR afterwards.Gossypium hirsutum L. Histone genes (GenBank accession
Number NC_006639) it is internal standard.Quantitative fluorescent PCR reaction kit is tried for the SYBR Premix Ex Taq II of TAKARA
Agent box, reactsCarry out in 96System fluorescent quantitation instruments, carry out the repetition of 3 secondary pollutants, experimental result phase
To quantitative 2ΔCtMethod calculates the expression (Schmittgen and Livak 2008) of GhNPC1b genes.Primer sequence is：
QGhNPC1b-F:TCCCCACTCTCTTGGTCTCTCG
QGhNPC1b-R:GAAGGGTAAGTATTGAGGACA
As a result：Expressions of the GhNPC1b in blade of a relatively high (Fig. 1).
Expression pattern analysis of the embodiment 4.GhNPC1b gene under abiotic stress
Gossypium hirsutum L. (kind be in 9807) is planted in greenhouse after seed is sterilized, and growth conditionss are：30 DEG C/25 DEG C, phase
It is 60-70% to humidity, illumination 14h, dark 10h, photon flux density are 800 μm of ol m-2s-1.When growing to four leaf stage, point
Low-phosphorous (5 μM), salt (200mM NaCl), the Stress treatment of arid (20%PEG6000) are not carried out, respectively in process 0,1,2,3,
Drawn materials when 6,9 and 12h, position is drawn materials for root.Extract total serum IgE and be reversed to cDNA, 10 times of dilution is used for glimmering in real time afterwards
Fluorescent Quantitative PCR is analyzed.Gossypium hirsutum L. Histone genes (GenBank accession number NC_006639) are internal standard.It is glimmering
SYBR Premix Ex Taq II test kit of the Fluorescent Quantitative PCR reaction kit for TAKARA, reacts
Carry out in 96System fluorescent quantitation instruments, carry out the repetition of 3 secondary pollutants, experimental result relative quantification 2-ΔΔCtMethod is calculated
The expression (Schmittgen and Livak 2008) of GhNPC1b genes.Primer sequence is：
QGhNPC1b-F:TCCCCACTCTCTTGGTCTCTCG
QGhNPC1b-R:GAAGGGTAAGTATTGAGGACA
As a result show, under low-phosphorus stress and salt stress, GhNPC1b expressions are raised, wherein, during low-phosphorus stress 6h,
The phenomenon for showing to lower of GhNPC1b；When drought stress processes 1h and 2h, GhNPC1b expressions are raised, table after 2h
Reveal the phenomenon of slight downward；Under ABA Stress treatments, GhNPC1b expressions are lowered.The expression of GhNPC1b genes by it is low-phosphorous,
The stress-inducing of salt and arid.
5. turns of GhNPC1b genes of embodiment create Resistance Strain of Cotton against material
One：The structure of pCAMBIA1300-GhNPC1b-EPSP plant expression vectors
When GhNPC1b overexpression structures are built, the cDNA nucleotide sequences of GhNPC1b are inserted into into plant expression vector
In pCAMBIA1300-EPSP, plasmid pCAMBIA1300-GhNPC1b-EPSP is obtained.After enzyme action identification is correct, plasmid is imported
In agrobacterium tumefaciens AGL1 or LBA4404, for Genetic Transformation in Higher Plants.Wherein plant expression vector pCAMBIA1300-EPSP is
Full synthetic EPSP genes (two ends add BamH I, KpnI restriction enzyme sites respectively) is linked to the multiple clone site of pCAMBIA1300
Upper acquisition.PCAMBIA1300 carriers in plant expression vector pCAMBIA1300-EPSP are CAMBIA Products,
GenBank number of registration AF134296；EPSP genes number of registration is AY573186.1.
Two：The acquisition of transgenic cotton plant
Cotton seeds soak 15min with the mercuric chloride of 70% ethanol immersion 1min, 0.1%, then with nothing Jing after sulphuric acid lint
Bacterium water washing 5 times.After sterilization, seed is put into and is covered with the aseptic bottle of three metafiltration paper, adds appropriate amounts of sterilized water, in 28 DEG C of incubators
Sprout.After 2～3 days, hypocotyl length is inserted into germination seed in MS solid mediums and continues culture to 1～2cm, and plant height 7～
It is used for converting during 10cm.
Will (plasmid pCAMBIA1300-GhNPC1b-EPSP carries herbicide resistance gene EPSP and purpose base with carrier
Because of GhNPC1b) agrobacterium tumefaciens (AGL1 or LBA4404) 28 DEG C in the YEP culture medium of additional antibiotic at concussion and cultivate,
Concussion speed is 110r/min, makes antibacterial be in exponential phase.Then it is centrifuged 10 minutes under 3000r/min, abandons supernatant.
Thalline is washed with 1/2MS improvement fluid mediums, is collected by centrifugation.Again by the thalline 1/2MS for adding 100mg/L acetosyringones
Improvement fluid medium suspends, and 5～20 times of dilution is used for converting.In conversion test, 1% surfactant Silwet is added
L-77 (Lehle Seeds, USA) or Tween80.
When kind of a skin comes off or comes off soon, remove kind of skin and a piece of cotyledon, expose seedling stem apex.If the existing lobule of shoot apex
Occur, then peelled off with tweezers.After gently scratching apical meristem, the cotton balls of Agrobacterium bacterium solution will be moistened with (by sterilized non-fat cotton
Prepare) seedling top is put into, cotton-wool is removed after continuing 24h, suck the residual bacterium solution of infection site with aseptic filter paper.Seedling is contaminated
When be placed in vacuum desiccator, be aided with 0.5 × 105The Negative pressure of Pa.The seedling contaminated in 21 DEG C or so light cultures 3 days,
Then flowerpot is transplanted into after cultivating 2～3 days under light.Flowerpot bottom is soil, and top is the thick Vermiculitums of 6～8cm, is poured every other day
1/2MS improves inorganic salt solution.
Three：Transformed plant is screened and field planting
After plant to be transformed grows 3 true leaves, 1.6～1.8 ‰ herbicide glyphosate (transgenic selectable marker genes are sprayed
For EPSP) aqueous solution, with unconverted plant as control.Fountain height falls drop with plant and is advisable.Unconverted plant 3 days after spraying
Stop growing, start within 7～10 days or so dead.Plant after conversion processing, the change of some individualities are similar to adjoining tree, separately
Some individualities then continued propagation, without significant change.Plant grew to for 4～5 leaf phases, was colonized to field.
Four：The identification and utilization of transfer-gen plant
The selfing of transfer-gen plant Post flowering or sisters' knot reality.Seed is sowed in land for growing field crops or greenhouse, in plant four leaf stage
Take blade and extract DNA, using round pcr detection progeny plant whether foreign gene-carrying GhNPC1b, and count exogenous gene and exist
Segregation ratio in filial generation.
As a result show, organize by transgene receptor of aseptic seedling shoot apical meristem, can effectively obtain transfer-gen plant.And
And, in partial transgenic strain offspring, exogenous gene mode of inheritance meets Mendel's law, and stablizes in passing on.Turn base
Because of homozygous line Jing field character determinations and resistant determination, select and keep receptor kind fundamental characteristics and resistance is significantly improved
Transgenic line, and improve for cotton variety.
The preferred embodiment of the present invention described in detail above.It should be appreciated that the ordinary skill of this area is without the need for wound
The property made work just can make many modifications and variations with design of the invention.Therefore, all technical staff in the art
Pass through the available technology of logical analysis, reasoning, or a limited experiment under this invention's idea on the basis of existing technology
Scheme, all should be in the protection domain being defined in the patent claims.
Sequence table
<110>Shandong University
<120>A kind of Gossypium hirsutum L. non-specificity phospholipase C gene GhNPC1b and its application
<141>2016-12-7
<160>2
<210>1
<211>1461
<212>cDNA
<213>Cotton Gossypii（Gossypiumspp）
<221>The cDNA nucleotide sequences of Gossypium hirsutum L. non-specificity phospholipase C gene GhNPC1b
<222>（1）…（1461）
<400>1
atggaaaacc gttcttttga ccatcttttg ggttggctca aatcgacccg acccgacata 60
gacggtctct ccggcacaga gtcaaaccca gtcaacgtcg ccgaccccaa ctcccccttt 120
atctctgtct ccgacgatgc tctctttgtc gactccgacc caggccactc ttttcaagcg 180
atcagagaac agattttcgg gtcgaacgac agctctgctg actcggctcc tatgaacggc 240
ttcgcccaac aagcggagag catgggtgaa ggaatgggta gaaccgtgat gagcggattt 300
aaaccgagtc ggttaccggt ttacacgaag ttagcgaacg agttcggcgt tctcgaccgg 360
tggtttgctt cggttccggc ttcgactcaa cctaacaggt tttacgttca ttcagcaacg 420
tcgtttgggg cgacgagtaa tgtcaagaaa gacctcatcc atggattccc ccaaaagacg 480
attttcgatt cgttggacga aaacggcctc agcttcggca tttattacca aaacatccct 540
gccacccttt tcttcaaaag cctaaggaaa ttaaagttct tgaccaaatt ccacaactac 600
gctttgaagt tccggctcca cgcgcggctt gggaagctgc cgaattacgt ggtggtggag 660
cagcgttact tcgacgtgaa ggagtttccg gcgaacgacg accacccgtc gcatgacgtg 720
gcgcgtgggc agaggttcgt gaaggaggtg tacgagatac tgagaagtag cccgcagtgg 780
aaagagatgg cgcttctgat cacgtacgat gagcacggag ggttttatga tcacgttccg 840
acacctgtct cgggtgttcc taacccggac ggaataattg gacccgaccc gttttatttc 900
aagttcaata ggcttggtgt tagggtcccc actctcttgg tctctcgctg gatcgataag 960
gcaactgtga tccacgagcc aactgggcca acaccgtctt cccaatttga acattcttcc 1020
atccctgcaa ctgtgaagaa gctcttcaac ctgaattcaa atttcctgac aaagagggat 1080
gcctgggctg ctacatttga aaattatttt aagctgcgta ctactccacg aactgactgt 1140
cctgaaactc ttccagaggt gacgacttca ttgaggccat gggggccaca agaagatgct 1200
agcctctcag aattccaagt tgagctggtt cagcttgcat cacagctcaa tggtgattat 1260
gtcctcaata cttacccttc tattgggaaa agcatgcgag taggtgaagc caaccgatac 1320
gtagaagatg cagtcaagag gttcctggaa gccggaaagg ctgctataag agccggagct 1380
aatgaatctg caattgttac aatgaggcct tctcttacca gtcgaatcga ggatcggagt 1440
caacatgtgg aagcctatta g 1461
<210>2
<211>486
<212>PRT
<213>Artificial sequence
<221>The aminoacid sequence of Gossypium hirsutum L. non-specificity phospholipase C gene GhNPC1b codings
<222>（1）…（486）
<400>2
MENRSFDHLL GWLKSTRPDI DGLSGTESNP VNVADPNSPF ISVSDDALFV DSDPGHSFQA 60
IREQIFGSND SSADSAPMNG FAQQAESMGE GMGRTVMSGF KPSRLPVYTK LANEFGVLDR 120
WFASVPASTQ PNRFYVHSAT SFGATSNVKK DLIHGFPQKT IFDSLDENGL SFGIYYQNIP 180
ATLFFKSLRK LKFLTKFHNY ALKFRLHARL GKLPNYVVVE QRYFDVKEFP ANDDHPSHDV 240
ARGQRFVKEV YEILRSSPQW KEMALLITYD EHGGFYDHVP TPVSGVPNPD GIIGPDPFYF 300
KFNRLGVRVP TLLVSRWIDK ATVIHEPTGP TPSSQFEHSS IPATVKKLFN LNSNFLTKRD 360
AWAATFENYF KLRTTPRTDC PETLPEVTTS LRPWGPQEDA SLSEFQVELV QLASQLNGDY 420
VLNTYPSIGK SMRVGEANRY VEDAVKRFLE AGKAAIRAGA NESAIVTMRP SLTSRIEDRS 480
QHVEAY 486
Claims (6)
1. a kind of Gossypium hirsutum L. non-specificity phospholipase C gene, it is characterised in that：The unnamed gene is Gossypium hirsutum L. non-specificity phosphorus
Lipase C gene GhNPC1b, as shown in SEQ ID No.1, the aminoacid sequence of its coding is such as the cDNA nucleotide sequences of the gene
Shown in SEQ ID No.2.
2. a kind of plant expression vector containing Gossypium hirsutum L. non-specificity phospholipase C gene GhNPC1b described in claim 1
pCAMBIA1300-GhNPC1b-EPSP。
3. applications of the Gossypium hirsutum L. non-specificity phospholipase C gene GhNPC1b described in claim 1 in Cotton Gossypii resistance is improved.
4. plant expression vector pCAMBIA1300-GhNPC1b-EPSP described in claim 2 improve Cotton Gossypii resistance in should
With.
5. the application as described in claim 3 or 4, it is characterised in that：The resistance refers to Salt And Alkali Tolerance, drought resisting and/or resistance to low
Phosphorus.
6. the application as described in claim 3 or 4, it is characterised in that：The cotton variety is spring cotton, summer cotton, normal conon or miscellaneous
Hand over cotton.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611213220.8A CN106520799A (en) | 2016-12-23 | 2016-12-23 | Non-specific phospholipase C gene GhNPC1b of upland cotton and applications of non-specific phospholipase C gene GhNPC1b |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611213220.8A CN106520799A (en) | 2016-12-23 | 2016-12-23 | Non-specific phospholipase C gene GhNPC1b of upland cotton and applications of non-specific phospholipase C gene GhNPC1b |
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| CN106520799A true CN106520799A (en) | 2017-03-22 |
Family
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115725615A (en) * | 2022-08-17 | 2023-03-03 | 甘肃农业大学 | Upland cotton GhABC1K14-A09 gene and application thereof in drought resistance and salt tolerance of upland cotton |
| CN117660523A (en) * | 2024-02-02 | 2024-03-08 | 河南大学三亚研究院 | Application of GhTSD7 gene in improving drought stress tolerance of plants |
-
2016
- 2016-12-23 CN CN201611213220.8A patent/CN106520799A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| CARLOTTA ANTONETTA PETERS: "Functional Characterization of Putative Non-Specific Phospholipase C (NPC) in Arabidopsis thaliana", 《DISSERTATIONS》 * |
| NCBI: "NCBI Reference Sequence：XM_016885196.1", 《NCBI》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115725615A (en) * | 2022-08-17 | 2023-03-03 | 甘肃农业大学 | Upland cotton GhABC1K14-A09 gene and application thereof in drought resistance and salt tolerance of upland cotton |
| CN117660523A (en) * | 2024-02-02 | 2024-03-08 | 河南大学三亚研究院 | Application of GhTSD7 gene in improving drought stress tolerance of plants |
| CN117660523B (en) * | 2024-02-02 | 2024-04-16 | 河南大学三亚研究院 | Application of GhTSD7 gene in improving drought stress tolerance of plants |
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