CN109852618A - A kind of section melon WRKY class transcription factor gene CqWRKY1 and its application - Google Patents
A kind of section melon WRKY class transcription factor gene CqWRKY1 and its application Download PDFInfo
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- CN109852618A CN109852618A CN201811541655.4A CN201811541655A CN109852618A CN 109852618 A CN109852618 A CN 109852618A CN 201811541655 A CN201811541655 A CN 201811541655A CN 109852618 A CN109852618 A CN 109852618A
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- cqwrky1
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- factor gene
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- FBLNYDYPCLFTSP-IXOXFDKPSA-N Ser-Phe-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O FBLNYDYPCLFTSP-IXOXFDKPSA-N 0.000 description 1
- DINQYZRMXGWWTG-GUBZILKMSA-N Ser-Pro-Pro Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 DINQYZRMXGWWTG-GUBZILKMSA-N 0.000 description 1
- FLONGDPORFIVQW-XGEHTFHBSA-N Ser-Pro-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CO FLONGDPORFIVQW-XGEHTFHBSA-N 0.000 description 1
- HHJFMHQYEAAOBM-ZLUOBGJFSA-N Ser-Ser-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O HHJFMHQYEAAOBM-ZLUOBGJFSA-N 0.000 description 1
- SRSPTFBENMJHMR-WHFBIAKZSA-N Ser-Ser-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SRSPTFBENMJHMR-WHFBIAKZSA-N 0.000 description 1
- JCLAFVNDBJMLBC-JBDRJPRFSA-N Ser-Ser-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JCLAFVNDBJMLBC-JBDRJPRFSA-N 0.000 description 1
- OSFZCEQJLWCIBG-BZSNNMDCSA-N Ser-Tyr-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O OSFZCEQJLWCIBG-BZSNNMDCSA-N 0.000 description 1
- UKKROEYWYIHWBD-ZKWXMUAHSA-N Ser-Val-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O UKKROEYWYIHWBD-ZKWXMUAHSA-N 0.000 description 1
- KEGBFULVYKYJRD-LFSVMHDDSA-N Thr-Ala-Phe Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 KEGBFULVYKYJRD-LFSVMHDDSA-N 0.000 description 1
- LVHHEVGYAZGXDE-KDXUFGMBSA-N Thr-Ala-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)N1CCC[C@@H]1C(=O)O)N)O LVHHEVGYAZGXDE-KDXUFGMBSA-N 0.000 description 1
- LIXBDERDAGNVAV-XKBZYTNZSA-N Thr-Gln-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O LIXBDERDAGNVAV-XKBZYTNZSA-N 0.000 description 1
- DJDSEDOKJTZBAR-ZDLURKLDSA-N Thr-Gly-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O DJDSEDOKJTZBAR-ZDLURKLDSA-N 0.000 description 1
- JKGGPMOUIAAJAA-YEPSODPASA-N Thr-Gly-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O JKGGPMOUIAAJAA-YEPSODPASA-N 0.000 description 1
- CYVQBKQYQGEELV-NKIYYHGXSA-N Thr-His-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N)O CYVQBKQYQGEELV-NKIYYHGXSA-N 0.000 description 1
- FWTFAZKJORVTIR-VZFHVOOUSA-N Thr-Ser-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O FWTFAZKJORVTIR-VZFHVOOUSA-N 0.000 description 1
- IVDFVBVIVLJJHR-LKXGYXEUSA-N Thr-Ser-Asp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O IVDFVBVIVLJJHR-LKXGYXEUSA-N 0.000 description 1
- KPMIQCXJDVKWKO-IFFSRLJSSA-N Thr-Val-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O KPMIQCXJDVKWKO-IFFSRLJSSA-N 0.000 description 1
- MVHHTXAUJCIOMZ-WDSOQIARSA-N Trp-Arg-Lys Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCCN)C(=O)O)N MVHHTXAUJCIOMZ-WDSOQIARSA-N 0.000 description 1
- JWGXUKHIKXZWNG-RYUDHWBXSA-N Tyr-Gly-Gln Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)NCC(=O)N[C@@H](CCC(=O)N)C(=O)O)N)O JWGXUKHIKXZWNG-RYUDHWBXSA-N 0.000 description 1
- NXRGXTBPMOGFID-CFMVVWHZSA-N Tyr-Ile-Asn Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O NXRGXTBPMOGFID-CFMVVWHZSA-N 0.000 description 1
- VTCKHZJKWQENKX-KBPBESRZSA-N Tyr-Lys-Gly Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O VTCKHZJKWQENKX-KBPBESRZSA-N 0.000 description 1
- GQVZBMROTPEPIF-SRVKXCTJSA-N Tyr-Ser-Asp Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O GQVZBMROTPEPIF-SRVKXCTJSA-N 0.000 description 1
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Abstract
The invention discloses a kind of section melon WRKY class transcription factor gene CqWRKY1, and nucleotide sequence is as shown in SEQ ID NO:1.And the coding albumen of section melon WRKY class transcription factor gene CqWRKY1, amino acid sequence is as shown in SEQ ID NO:2.The invention also discloses application of the above-mentioned section melon WRKY class transcription factor gene CqWRKY1 in terms of regulation saves melon fusaric acid resistance.
Description
Technical field
The invention belongs to save melon technical field, and in particular to a kind of section melon WRKY class transcription factor gene CqWRKY1 and its
Using.
Background technique
Cucurbits fusarium wilt (FusarumoxysporusSchl.f.sp.cucumerirum Owen.) is also known as dead arm, withers
It withers disease, is a kind of worldwide soil-borne disease, is generally caused by Fusarium oxysporum (Fusariumoxysporum), main harm is yellow
Other Curcurbitaceae category crops such as melon, wax gourd, section melon.
Section melon (Benincasa hispida Cogn.var.Chieh-qua How) is the important vegetable of Curcurbitaceae Benincasa
Dish crop and the famous-brand and high-quality melon vegetables of China's Lingnan area, are loved by consumers.In recent years, the morbidity of section cucurbit wilt becomes
Gesture is increasingly sharpened, and the production of section melon is seriously hindered.Since section melon genetic background is narrow, anti-blight resource is less, not yet divides
Excellent fusarium wilt disease resistance gene is separated out, and resistance molecule mechanism study lacks.
WRKY class transcription factor is the important transcription regulatory factor of one kind being widely present in plant kingdom, and N- contains at end
The highly conserved amino acid sequence of WRKYGQK participates in plant pair by adjusting the expression of the gene of the element containing W-box in promoter
The processes such as defense response, abiotic stress, the growth and development of pathogen especially play important work in Plant defense responses
With.However, there is not been reported for the research about WRKY class transcription factor in terms of saving cucurbit wilt resistance at present.Therefore, carry out
Research of the melon WRKY class transcription factor in terms of saving cucurbit wilt resistance is saved, WRKY ginseng can be not only disclosed in terms of genetic level
With the effect of section cucurbit wilt resistance, theory support also is provided for section cucurbit wilt resistance molecule assistant breeding, there is important grind
Study carefully value.
Summary of the invention
The purpose of the present invention is to provide a kind of section melon WRKY class transcription factor gene CqWRKY1 and its coding albumen.
The object of the invention is also to provide the applications of above-mentioned section melon WRKY class transcription factor gene CqWRKY1.
Above-mentioned first purpose of the invention is achieved through the following technical solutions: a kind of transcription of section melon WRKY class because
Subbase is because of CqWRKY1, and nucleotide sequence is as shown in SEQ ID NO:1.
The coding albumen of above-mentioned section melon WRKY class transcription factor gene CqWRKY1, amino acid sequence such as SEQ ID NO:2
It is shown.
Section melon WRKY class transcription factor gene CqWRKY1 of the present invention is the total serum IgE reverse transcription that will save melon seedling true leaf
At cDNA, using cDNA as template, PCR amplification acquisition is carried out with the primer pair as shown in SEQ ID NO:3 and SEQ ID NO:4.
Wherein primer pair specifically:
CqWRKY1-F:ATGGCCTCCTCTTCCGGGAG
CqWRKY1-R:CTAACATAGGAGAGATTGGA
Above-mentioned second purpose of the invention is achieved through the following technical solutions: above-mentioned section melon WRKY class transcription
Application of the factor gene CqWRKY1 in terms of regulation saves melon fusaric acid resistance.
The present invention obtains pEarleyGate101- in CqWRKY1 gene cloning to 101 carrier of pEarleyGate
CqWRKY1 plant over-express vector.
As a result, it has been found that turning base under fusaric acid (Fusaricacid, FA, Sclerotium rolfsii secretion are commercially available) stress
Because the resistance of plant is markedly less than control, illustrate that save melon WRKY class transcription factor gene CqWRKY1 resists in regulation section cucurbit wilt
Property aspect have certain negative role.Therefore, it in subsequent production practices, can be transcribed by knocking out the WRKY class in section melon
Factor gene CqWRKY1, to improve the anti-blight performance of section melon.
The present invention has the advantage that the present invention identifies the coding of section melon CqWRKY1 using the separation of electronic cloning technology
Sequence information；Have studied expression of the CqWRKY1 under different environment stresses using special primer, discovery section melon WRKY class transcription because
Subbase can be by various abiotic stress factor inducing expression because of CqWRKY1；The section melon WRKY class transcription factor gene CqWRKY1 mistake of building
The fusaric acid resistance for expressing plant reduces, and illustrates the fusarium wilt disease resistance of the gene negative regulation section melon, thus can further exist
It, can be by knocking out the WRKY class transcription factor gene CqWRKY1 in section melon, to improve section melon in subsequent production practices
Anti-blight performance.
Detailed description of the invention
Fig. 1 is the clone that melon CqWRKY1 gene is saved in embodiment 1, and wherein 1-3 is the CDS amplification knot of CqWRKY1 gene
Fruit, M are DNA Marker bands；
Fig. 2 is that expression of the melon CqWRKY1 gene under different environment stresses is saved in embodiment 2；
Fig. 3 is the detection for saving melon CqWRKY1 in embodiment 3 and being overexpressed positive plant；
Fig. 4 is that the fusaric acid resistance that CqWRKY1 is overexpressed plant in embodiment 4 reduces, and wherein A figure represents fusaric acid
The upgrowth situation of plant before handling, B figure are the growing state of plant after fusaric acid processing, and OE5,6,13 respectively represent overexpression
The positive homozygous lines of (Over expressing:OE) CqWRKY1 gene.
Specific embodiment
A specific embodiment of the invention is specifically further illustrated below with reference to example.
1 section melon WRKY class transcription factor gene CqWRKY1 of embodiment and its acquisition for encoding albumen
(1) RNA extracts the clone with cDNA
The section melon kind of selection is the homozygosis that the spontaneous pollination offspring of new farmer section melon kind comes by pedigree method breeding
It is selfed based material.
It hots water treatment of seeds seed 6-8 hours, then vernalization 2-3 days in 30-32 DEG C of incubator, is broadcast when seed shows money or valuables one carries unintentionally
Kind.After seedling grows to a leaf wholeheartedly, mixed sampling is carried out to the true leaf of 3 plants of seedling, be added after liquid nitrogen with mortar cracking tissue and
Cell, and be transferred to rapidly in the centrifuge tube of 2.0mL, using plant total RNA extraction reagent box (RNAprep pure Tissue
Kit, TIANGEN)；The concentration of RNA, while the integrality of agarose gel electrophoresis detection RNA are detected with spectrophotometer.It connects down
Come, using Dalian treasured biotech firm Reverse Transcriptase kit (PrimeScript Reverse Transcriptase kit,
Takara), reverse transcription is at cDNA.
(2) amplification of CqWRKY1 gene
To save the total cDNA of melon as template, with CqWRKY1-F (atggcctcctcttccgggag, such as SEQ ID NO:3 institute
Show) and CqWRKY1-R (ctaacataggagagattgga, as shown in SEQ ID NO:4) obtained for positive and reverse primer, clone
Melon CqWRKY1 genetic fragment must be saved.
PCR response procedures are as follows: 94 DEG C initial denaturation 5 minutes；94 DEG C be denaturalized 30 seconds, 58 DEG C renaturation 2 minutes, 72 DEG C extend 45
Second, 72 DEG C 8 minutes after 34 circulations.Final PCR product is sequenced, and the sequence of CqWRKY1 is obtained, as shown in Figure 1, wherein
1-3 is the CDS amplification of CqWRKY1 gene, and M is DNA Marker band.
It is compared and is found by NCBI-Blast and MEGA6.0 software, the sequence and watermelon WRKY1-like
(HM030879.1), the homologys such as cucumber WRKY33-like (NM_001280728.1) are higher, and contain PolyA structure, show
CqWRKY1 is the gene of a coding WRKY class transcription factor.
The nucleotide sequence of section melon WRKY class transcription factor gene CqWRKY1 is specific as follows:
atggcctcctcttccgggagcgtagacacctctgctaattctcatccttctttcactttctctactca
tccttttatgacttcttactctgaccttctcgcttcctccaacaacgatcctccttcctccgccgcaccccacgct
tctctccgcggttccggtactggggttcctaaattcaaatccctccctcctccttctctccctctctctcctcctc
ccatgtctccttcttcctttttcgctattcctcctggcttgagtcccgctgagcttcttgattcccctgttcttct
cagtgcttctcatgttctgccgtcgccgactaccgggagtttcccgtctcagtcgttgaattggaagagcaattct
gggtgtaatcagcagagcattaaggaagaaaacaaatatttgtccaatttctcatttcaaactcaatcgtcaaagt
ttccgccgacgtcttttcagccttcatccaccacagctcctacgactcagggatggagttttcaagaacagcggaa
gaaagaggacggtttctcgtcggagaagaatatggtgaagccggagttcggatcgatgcggagcttctcaccggaa
tatggggttgttcaaaaccagagccaaaacaacggcagcggggagttgcagtctgactacggcaataattaccctc
aacaatctcagacgttgaatcgaaggtcggacgacggctacaactggagaaaatacggccaaaaacaggttaaagg
aagtgaaaatccgagaagctattataagtgcactttccccaattgcccaactaagaaaaaggttgaaagatcctta
gatggacagatcacggagattgtttacaaaggcagccataaccatgccaagccccaatccacgaggaggtcgtcac
tgtcgtcagccggttcttcccacgccctggtggctttgattccggctactaatgagatggcggaccaatcatttac
aacccaaggcagcggccaattcgacggcgttgcaacgccggagaattcctcaatttcaatcggcgacgacgacttc
gatcgaagctctcaaaagagcaaatccggaggggacgattttgatgaggacgaaccagaggctaagagatggcgaa
gggaaggtgacaacaatgaaggtatttcggcggtcggtagccggacggtgagagagccgagagtcgtcgtccaaac
caccagcgacatcgacattctggacgatggttaccggtggaggaagtacggccagaaagttgtgaagggaaaccca
aatccaaggagttactacaaatgcacaaatccaggatgtccagtaagaaagcacgtggagagagcttcacatgatc
taagagcagtgatcacaacttatgaagggaagcacaaccatgatgttccaccagcacgtggcagtggaagccattc
cctcagccgtccattccccagcaacgaccctccggccacggcgatccgcccatcggcagtgacccatcagtcaaac
aacggggggcatttgcaaggtctaaggctgcagcaatcttcagagtcccaaacagcattcacagtggaaatggtgc
aaaatgggaatggattttcattcccagaatttggaaactcaatgggaatgggagtaggatcctacatcaaccaaac
acagcctaatgacaatttgttaatcagagctaaagaagagccaagagatcatgacatgttcatccaatctctccta
tgttag。
The amino acid sequence of the coding albumen of section melon WRKY class transcription factor gene CqWRKY1 is specific as follows:
MASSSGSVDTSANSHPSFTFSTHPFMTSYSDLLASSNNDPPSSAAPHASLRGSGTGVPKFKSLPPPSL
PLSPPPMSPSSFFAIPPGLSPAELLDSPVLLSASHVLPSPTTGSFPSQSLNWKSNSGCNQQSIKEENKYLSNFSFQ
TQSSKFPPTSFQPSSTTAPTTQGWSFQEQRKKEDGFSSEKNMVKPEFGSMRSFSPEYGVVQNQSQNNGSGELQSDY
GNNYPQQSQTLNRRSDDGYNWRKYGQKQVKGSENPRSYYKCTFPNCPTKKKVERSLDGQITEIVYKGSHNHAKPQS
TRRSSLSSAGSSHALVALIPATNEMADQSFTTQGSGQFDGVATPENSSISIGDDDFDRSSQKSKSGGDDFDEDEPE
AKRWRREGDNNEGISAVGSRTVREPRVVVQTTSDIDILDDGYRWRKYGQKVVKGNPNPRSYYKCTNPGCPVRKHVE
RASHDLRAVITTYEGKHNHDVPPARGSGSHSLSRPFPSNDPPATAIRPSAVTHQSNNGGHLQGLRLQQSSESQTAF
TVEMVQNGNGFSFPEFGNSMGMGVGSYINQTQPNDNLLIRAKEEPRDHDMFIQSLLC-。
Expression study of the embodiment 2CqWRKY1 under different environment stresses
According to the cDNA sequence of CqWRKY1, real-time quantitative PCR (qRT-PCR) primer: CqWRKY1-Q-F is designed:
CCTCCTCCTTCTCTCCCTCT；CqWRKY1-Q-R:AACGACTGAGACGGGAAACT.Internal control primer used is section melon
Actin primer, Actin-Q-F:TCAACCCAAAGGCTAACAG；Actin-Q-R:CTTGTCCATCAGGCAGTTC.
CDNA template is diluted 5 times, prepares the reaction system of 20 μ L: 10 μ L SYBR Premix Ex Taq II (2 ×),
0.8 μ LForward Primer, 0.8 μ LReverse Primer, 2 μ LcDNA, 0.4 μ LRox Reference Dye II, 6.0
μl ddH2Then O carries out expanding on fluorescence quantitative PCR instrument (ABI Prism 7500HT, Applied Biosystems) anti-
It answers.
Response procedures are as follows: 95 DEG C, 30sec, 95 DEG C, 5sec, 60 DEG C, 34sec, 40 recycle in total.Then fixed in fluorescence
It is carried out amplification reaction in amount PCR instrument (ABI Prism 7500HT, Applied Biosystems), in Ubiqutin gene work
Ginseng, calculates the relative expression quantity of gene.
Each sample is arranged 3 biology and repeats, and each biology repeats to include that 3 technologies repeat quantitatively.Respectively just
Under the conditions of often, ABA, 6-BA, GA, IAA, high temperature detect the expression quantitative change of CqWRKY1 under the conditions of low temperature and fusaric acid (FA)
Change.
As a result as shown in Fig. 2, CqWRKY1 is most expressed vulnerable to FA stress-inducing as can be drawn from Figure 2.
The building of embodiment 3CqWRKY1 over-express vector and the detection of positive plant
(1) building of CqWRKY1 over-express vector
Using the cDNA of CqWRKY1 gene as template, the code area of PCR amplification 1746bp, pcr amplification product recovery purifying,
CqWRKY1 gene is connected on 101 plasmid of over-express vector pEarleyGate using Gateway technology, is selected after conversion
Positive colony is sequenced, and correct plasmid converts Agrobacterium GV3101, using the Kan of the Rif of 50mg/L and 50mg/L as resisting
Property screening, the monoclonal of acquisition is accredited as the positive by PCR.Pass through agriculture stalk bacterium mediated method, conversion section melon material.
(2) it is overexpressed plant detection
According to over-express vector sequence, (101 carrier of pEarleyGate is by Vegetables Inst., Guangdong Academy of Agricultural Sciences
There is provided, for preparing over-express vector in (1)) and CqWRKY1 code area (CDS) sequence design specific primer
CqWRKY1-F:ATGGCCTCCTCTTCCGGGAG；CqWRKY1-R:CTAACATAGGAGAGATTGGA)).To convert blank load
Body is control, and pcr amplification product is detected with 1% agarose electrophoresis, and can amplify target stripe is positive plant；Another party
Face, is control with the expression quantity of the CqWRKY1 gene of transformation receptor, and CqWRKY1 gene expression dose significantly rises
Plant is positive plant, the respectively positive homozygous lines OE5 of overexpression (Over expressing:OE) CqWRKY1 gene,
OE6 and OE13 (Fig. 3).
Embodiment 4 is overexpressed the fusaric acid stress resistance identification of plant
By the T2 pure lines hairy squash seed for being overexpressed CqWRKY1 gene, (T0 obtains T1 generation by pollination self for positive plant
Plant, then in T1 generation, obtains T2 for plant by pollination self, detects acquisition pure lines by qPCR and is overexpressed seed) and control kind
Son carries out vernalization under the same conditions, is sowed in the matrix of sterilizing later.When growing to a leaf wholeheartedly, seedling is removed into matrix
And root soil is cleaned up, it is put it into culture bottle after scratching root with blade, the reaping hook that concentration is 200mg/L is added
Bacterium acid solution (Fusaricacid, FA, Sclerotium rolfsii secretion are commercially available), makes seedling root be completely submerged in fusaric acid molten
In liquid.The growing state of plant is observed at any time, until difference occurs between control and transgenic plant in phenotype.In each culture bottle
6 plants are put, does 3 repetitions in parallel every time.
Figure 4, it is seen that the resistance of transgenic plant is markedly less than control, illustrates that CqWRKY1 exists under FA stress
There is certain negative role in terms of regulation section cucurbit wilt resistance.But it can be knocked out by gene editing technology in production
CqWRKY1 gene, or by RNA perturbation technique, the expression quantity of CqWRKY1 gene is reduced, to reach raising section cucurbit wilt
The effect of resistance.
The above is only non-limiting embodiment of the invention, for those of ordinary skill in the art, not
Under the premise of being detached from the invention design and not making creative work, various modifications and improvements can be made, these are all
It belongs to the scope of protection of the present invention.
Sequence table
<110>Vegetables Inst., Guangdong Academy of Agricultural Sciences
<120>a kind of section melon WRKY class transcription factor gene CqWRKY1 and its application
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1746
<212> DNA
<213>melon (Benincasa hispida Cogn. var. Chieh-qua How) is saved
<400> 1
atggcctcct cttccgggag cgtagacacc tctgctaatt ctcatccttc tttcactttc 60
tctactcatc cttttatgac ttcttactct gaccttctcg cttcctccaa caacgatcct 120
ccttcctccg ccgcacccca cgcttctctc cgcggttccg gtactggggt tcctaaattc 180
aaatccctcc ctcctccttc tctccctctc tctcctcctc ccatgtctcc ttcttccttt 240
ttcgctattc ctcctggctt gagtcccgct gagcttcttg attcccctgt tcttctcagt 300
gcttctcatg ttctgccgtc gccgactacc gggagtttcc cgtctcagtc gttgaattgg 360
aagagcaatt ctgggtgtaa tcagcagagc attaaggaag aaaacaaata tttgtccaat 420
ttctcatttc aaactcaatc gtcaaagttt ccgccgacgt cttttcagcc ttcatccacc 480
acagctccta cgactcaggg atggagtttt caagaacagc ggaagaaaga ggacggtttc 540
tcgtcggaga agaatatggt gaagccggag ttcggatcga tgcggagctt ctcaccggaa 600
tatggggttg ttcaaaacca gagccaaaac aacggcagcg gggagttgca gtctgactac 660
ggcaataatt accctcaaca atctcagacg ttgaatcgaa ggtcggacga cggctacaac 720
tggagaaaat acggccaaaa acaggttaaa ggaagtgaaa atccgagaag ctattataag 780
tgcactttcc ccaattgccc aactaagaaa aaggttgaaa gatccttaga tggacagatc 840
acggagattg tttacaaagg cagccataac catgccaagc cccaatccac gaggaggtcg 900
tcactgtcgt cagccggttc ttcccacgcc ctggtggctt tgattccggc tactaatgag 960
atggcggacc aatcatttac aacccaaggc agcggccaat tcgacggcgt tgcaacgccg 1020
gagaattcct caatttcaat cggcgacgac gacttcgatc gaagctctca aaagagcaaa 1080
tccggagggg acgattttga tgaggacgaa ccagaggcta agagatggcg aagggaaggt 1140
gacaacaatg aaggtatttc ggcggtcggt agccggacgg tgagagagcc gagagtcgtc 1200
gtccaaacca ccagcgacat cgacattctg gacgatggtt accggtggag gaagtacggc 1260
cagaaagttg tgaagggaaa cccaaatcca aggagttact acaaatgcac aaatccagga 1320
tgtccagtaa gaaagcacgt ggagagagct tcacatgatc taagagcagt gatcacaact 1380
tatgaaggga agcacaacca tgatgttcca ccagcacgtg gcagtggaag ccattccctc 1440
agccgtccat tccccagcaa cgaccctccg gccacggcga tccgcccatc ggcagtgacc 1500
catcagtcaa acaacggggg gcatttgcaa ggtctaaggc tgcagcaatc ttcagagtcc 1560
caaacagcat tcacagtgga aatggtgcaa aatgggaatg gattttcatt cccagaattt 1620
ggaaactcaa tgggaatggg agtaggatcc tacatcaacc aaacacagcc taatgacaat 1680
ttgttaatca gagctaaaga agagccaaga gatcatgaca tgttcatcca atctctccta 1740
tgttag 1746
<210> 2
<211> 581
<212> PRT
<213>melon (Benincasa hispida Cogn. var. Chieh-qua How) is saved
<400> 2
Met Ala Ser Ser Ser Gly Ser Val Asp Thr Ser Ala Asn Ser His Pro
1 5 10 15
Ser Phe Thr Phe Ser Thr His Pro Phe Met Thr Ser Tyr Ser Asp Leu
20 25 30
Leu Ala Ser Ser Asn Asn Asp Pro Pro Ser Ser Ala Ala Pro His Ala
35 40 45
Ser Leu Arg Gly Ser Gly Thr Gly Val Pro Lys Phe Lys Ser Leu Pro
50 55 60
Pro Pro Ser Leu Pro Leu Ser Pro Pro Pro Met Ser Pro Ser Ser Phe
65 70 75 80
Phe Ala Ile Pro Pro Gly Leu Ser Pro Ala Glu Leu Leu Asp Ser Pro
85 90 95
Val Leu Leu Ser Ala Ser His Val Leu Pro Ser Pro Thr Thr Gly Ser
100 105 110
Phe Pro Ser Gln Ser Leu Asn Trp Lys Ser Asn Ser Gly Cys Asn Gln
115 120 125
Gln Ser Ile Lys Glu Glu Asn Lys Tyr Leu Ser Asn Phe Ser Phe Gln
130 135 140
Thr Gln Ser Ser Lys Phe Pro Pro Thr Ser Phe Gln Pro Ser Ser Thr
145 150 155 160
Thr Ala Pro Thr Thr Gln Gly Trp Ser Phe Gln Glu Gln Arg Lys Lys
165 170 175
Glu Asp Gly Phe Ser Ser Glu Lys Asn Met Val Lys Pro Glu Phe Gly
180 185 190
Ser Met Arg Ser Phe Ser Pro Glu Tyr Gly Val Val Gln Asn Gln Ser
195 200 205
Gln Asn Asn Gly Ser Gly Glu Leu Gln Ser Asp Tyr Gly Asn Asn Tyr
210 215 220
Pro Gln Gln Ser Gln Thr Leu Asn Arg Arg Ser Asp Asp Gly Tyr Asn
225 230 235 240
Trp Arg Lys Tyr Gly Gln Lys Gln Val Lys Gly Ser Glu Asn Pro Arg
245 250 255
Ser Tyr Tyr Lys Cys Thr Phe Pro Asn Cys Pro Thr Lys Lys Lys Val
260 265 270
Glu Arg Ser Leu Asp Gly Gln Ile Thr Glu Ile Val Tyr Lys Gly Ser
275 280 285
His Asn His Ala Lys Pro Gln Ser Thr Arg Arg Ser Ser Leu Ser Ser
290 295 300
Ala Gly Ser Ser His Ala Leu Val Ala Leu Ile Pro Ala Thr Asn Glu
305 310 315 320
Met Ala Asp Gln Ser Phe Thr Thr Gln Gly Ser Gly Gln Phe Asp Gly
325 330 335
Val Ala Thr Pro Glu Asn Ser Ser Ile Ser Ile Gly Asp Asp Asp Phe
340 345 350
Asp Arg Ser Ser Gln Lys Ser Lys Ser Gly Gly Asp Asp Phe Asp Glu
355 360 365
Asp Glu Pro Glu Ala Lys Arg Trp Arg Arg Glu Gly Asp Asn Asn Glu
370 375 380
Gly Ile Ser Ala Val Gly Ser Arg Thr Val Arg Glu Pro Arg Val Val
385 390 395 400
Val Gln Thr Thr Ser Asp Ile Asp Ile Leu Asp Asp Gly Tyr Arg Trp
405 410 415
Arg Lys Tyr Gly Gln Lys Val Val Lys Gly Asn Pro Asn Pro Arg Ser
420 425 430
Tyr Tyr Lys Cys Thr Asn Pro Gly Cys Pro Val Arg Lys His Val Glu
435 440 445
Arg Ala Ser His Asp Leu Arg Ala Val Ile Thr Thr Tyr Glu Gly Lys
450 455 460
His Asn His Asp Val Pro Pro Ala Arg Gly Ser Gly Ser His Ser Leu
465 470 475 480
Ser Arg Pro Phe Pro Ser Asn Asp Pro Pro Ala Thr Ala Ile Arg Pro
485 490 495
Ser Ala Val Thr His Gln Ser Asn Asn Gly Gly His Leu Gln Gly Leu
500 505 510
Arg Leu Gln Gln Ser Ser Glu Ser Gln Thr Ala Phe Thr Val Glu Met
515 520 525
Val Gln Asn Gly Asn Gly Phe Ser Phe Pro Glu Phe Gly Asn Ser Met
530 535 540
Gly Met Gly Val Gly Ser Tyr Ile Asn Gln Thr Gln Pro Asn Asp Asn
545 550 555 560
Leu Leu Ile Arg Ala Lys Glu Glu Pro Arg Asp His Asp Met Phe Ile
565 570 575
Gln Ser Leu Leu Cys
580
<210> 3
<211> 20
<212> DNA
<213>melon (Benincasa hispida Cogn. var. Chieh-qua How) is saved
<400> 3
atggcctcct cttccgggag 20
<210> 4
<211> 20
<212> DNA
<213>melon (Benincasa hispida Cogn. var. Chieh-qua How) is saved
<400> 4
ctaacatagg agagattgga 20
Claims (4)
1. a kind of section melon WRKY class transcription factor gene CqWRKY1, it is characterized in that: its nucleotide sequence such as SEQ ID NO:1 institute
Show.
2. the coding albumen of melon WRKY class transcription factor gene CqWRKY1 is saved described in claim 1, it is characterized in that: its amino acid
Sequence is as shown in SEQ ID NO:2.
3. section melon WRKY class transcription factor gene CqWRKY1 according to claim 1, it is characterized in that: the section melon WRKY
Class transcription factor gene CqWRKY1 is that will save the total serum IgE reverse transcription of melon seedling true leaf at cDNA, using cDNA as template, with such as SEQ
Primer pair shown in ID NO:3 and SEQ ID NO:4 carries out PCR amplification acquisition.
4. section melon WRKY class transcription factor gene CqWRKY1 described in claim 1 is in terms of regulation saves melon fusaric acid resistance
Application.
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Cited By (5)
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| CN111217896A (en) * | 2020-02-28 | 2020-06-02 | 天津大学 | Application of ginseng PgWRKY4X transcription factor in regulating and controlling ginsenoside compound content in ginseng |
| CN111548398A (en) * | 2020-05-25 | 2020-08-18 | 海南大学 | Anthrax bacterium transcription factor CsATF1 and application |
| CN111635904A (en) * | 2020-06-29 | 2020-09-08 | 山东农业大学 | A gene CsWRKY10 enhancing resistance to target spot disease of cucumber and its application |
| CN112341530A (en) * | 2020-11-23 | 2021-02-09 | 中国科学院华南植物园 | Dendrobium WRKY transcription factor gene DoWRKY69 and application thereof |
| CN117625684A (en) * | 2023-12-06 | 2024-03-01 | 河北省农林科学院经济作物研究所 | Application of SmWRKY33 protein of red sage root and its coding gene in regulating and controlling salt tolerance of plant |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111217896A (en) * | 2020-02-28 | 2020-06-02 | 天津大学 | Application of ginseng PgWRKY4X transcription factor in regulating and controlling ginsenoside compound content in ginseng |
| CN111217896B (en) * | 2020-02-28 | 2022-04-29 | 天津大学 | Application of ginseng PgWRKY4X transcription factor in regulating the content of ginsenosides in ginseng |
| CN111548398A (en) * | 2020-05-25 | 2020-08-18 | 海南大学 | Anthrax bacterium transcription factor CsATF1 and application |
| CN111548398B (en) * | 2020-05-25 | 2022-02-11 | 海南大学 | An anthracis transcription factor CsATF1 and its application |
| CN111635904A (en) * | 2020-06-29 | 2020-09-08 | 山东农业大学 | A gene CsWRKY10 enhancing resistance to target spot disease of cucumber and its application |
| CN112341530A (en) * | 2020-11-23 | 2021-02-09 | 中国科学院华南植物园 | Dendrobium WRKY transcription factor gene DoWRKY69 and application thereof |
| CN117625684A (en) * | 2023-12-06 | 2024-03-01 | 河北省农林科学院经济作物研究所 | Application of SmWRKY33 protein of red sage root and its coding gene in regulating and controlling salt tolerance of plant |
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Application publication date: 20190607 |