Application of methyltransferase gene EFS in regulation and control of plant salt stress resistance

Abstract

The invention discloses application of a methyltransferase gene EFS in regulation and control of plant salt stress resistance, and belongs to the technical field of plant molecular biology. The methyltransferase gene EFS is separated from arabidopsis thaliana, the full-length CDS of the methyltransferase gene EFS is 5418bp, the methyltransferase gene EFS has three conservative protein function structural domains which are respectively an SET structural domain, an AWS structural domain and a ZnfCW structural domain, and the protein coded by the methyltransferase gene EFS contains 1805 amino acids. After the methyltransferase gene EFS is knocked out, 100mM NaCl and 125mM NaCl are used for treating a mutant, so that the sensitivity of the mutant to the salt stress is increased, which indicates that the mutant plays an important role in the response of the plant to the salt stress, and therefore, the gene can be applied to the salt stress resistance of the plant.

Description

Technical Field

[0001] The present invention relates to the technical field of plant molecular biology, and particularly relates to the application of a methyltransferase gene EFS in regulating plant salt stress resistance. Background Art

[0002] Salt stress occurs when plants grow in a high-salt environment and are affected by high osmotic potential, and its essence is osmotic stress. High salt concentration can cause plants to be unable to effectively absorb water at the roots, resulting in a "salt osmosis" effect. The increase in salt content around the roots will cause plants to lose water, thus triggering water stress and affecting the normal growth of plants. After sodium ions (Na+) and chloride ions (Cl-) in the salt accumulate in plants, they will be toxic to plant cells, interfere with the stability of cell membranes, affect the activity of enzymes, and damage the physiological functions of plants. Salt stress usually significantly reduces the growth rate, root development, and yield of plants. Under natural conditions, plants are mainly stressed by sodium salts. Existing genes that can regulate plant salt stress resistance include: GhWRKY41 gene in cotton, GmNHX1 gene in soybean, and TaNHX1-2 gene in wheat, etc., but it has not been reported that the methyltransferase gene EFS can regulate plant salt stress resistance. Summary of the Invention

[0003] In order to solve the above-mentioned deficiencies existing in the prior art, the purpose of the present invention is to provide the application of a methyltransferase gene EFS in regulating plant salt stress resistance.

[0004] The technical solution of the present invention to solve the above technical problems is as follows: Provide the application of a methyltransferase gene EFS in regulating plant salt stress resistance, wherein the nucleotide sequence of the methyltransferase gene EFS is as shown in SEQ ID NO.1.

[0005] Further, the plant is Arabidopsis thaliana.

[0006] Further, knocking out the methyltransferase gene EFS reduces the salt stress resistance performance of plants.

[0007] A preparation for regulating plant salt stress resistance includes the above-mentioned methyltransferase gene EFS or the protein encoded by this gene.

[0008] A method for regulating plant salt stress resistance includes the following steps:

[0009] Regulating the salt stress resistance performance of plants by knocking out or overexpressing the methyltransferase gene EFS.

[0010] The present invention has the following beneficial effects:

[0011] The methyltransferase gene EFS in the present invention was isolated from Arabidopsis thaliana. Its full-length CDS is 5418 bp, and it has three conserved protein functional domains, namely the SET domain, the AWS domain, and the Znf_CW domain. The protein encoded by it contains 1805 amino acids.

[0012] After knocking out the methyltransferase gene EFS and treating the mutants with 100 mM NaCl and 125 mM NaCl, the sensitivity of the mutants to salt stress increased, indicating that it plays an important role in the response of plants to salt stress. Therefore, this gene can be applied to the application of plant salt stress resistance. Description of the Drawings

[0013] Figure 1 It is the phenotypic result of the EFS gene knockout mutant after salt stress treatment. Detailed Embodiments

[0014] The following examples are only used to explain the present invention and are not intended to limit the scope of the present invention. For those not specified in the examples, they are carried out according to the conventional conditions or the conditions recommended by the manufacturer. For the reagents or instruments not specified for the manufacturer, they are all conventional products that can be obtained through commercial purchase.

[0015] Example 1:

[0016] The methyltransferase gene EFS in the present invention was isolated from Arabidopsis thaliana. Its full-length CDS is 5418 bp, and the nucleotide sequence is shown in SEQ ID NO.1. The protein encoded by it contains 1805 amino acids, and the specific sequence is shown in SEQ ID NO.2.

[0017] ATGGACTGTAAGGAAAACGGTGTTGGTGACGCTTCTGGGTGCAATATTGATGCTAACTCTCTGGCCTCAAATCTTGCGATGAATACCAATGAAGATTTCTATGAGAAATTGAGTTCTCGTGGGCAAAATTTAGATTCAGTCAGTAGTTTGGAGATTCCTCAGACTGCTTCATCTGTAAACCATACGATTGAAGGCCAAAGGAAGTGCTTTACAGAGATTGAGCAGATGGGATACGGGAACAGTAATAGTCAAGAAGATGCTGGAAACACTGATGATGATCTATATGTTTGTTACAATGCTGATGACACTCAGGAGCAGGGAGTGGTTTCAGGTGAGCTTGAACAAAGTCAAGAATTAATTTGTGATACTGATTTACTGGTAAATTGCAACAAGCTGGATGATGGAAAGGAAAGTCAGGACACAAATGTGTCCCTTGTATCCATTTTTTCTGGGAGTATGCAAGAGAAAGAAG

[0018] CTCCTCAG

[0019] GCTAAAGAAGATGAAGGTTATGGTGGCACAACATTGCCTATAGGGGG

[0020] CAGTGGAATAGATACGGAATCAACTTTTGTGAATGATGCACCAGAAC

[0021] AATTCGAGTCTTTGGAAACCACAAAGCACATAAAACCTGATGAAGTT

[0022] GAAAGTGATGGTATTAGTTATAGGTTTGATGATGGAGGTAAGGAAGG

[0023] AAGAAATGGACCATCGAGTGATCTAGATACTGGTTCTTCTGATGATAT

[0024] ATCTCTCAGTCAGAGTTTTTCGTTTCCAGATTCGTTATTGGACTCTAGT

[0025] GTATTTGGCTGTAGCGCCACAGAAAGTTATCTGGAGGATGCAATTGAT

[0026] ATTGAAGGCAATGGGACTATTGTTGTATCCCCTTCATTAGCTATCACA

[0027] GAAATGTTAAATAATGATGATGGTGGCCTATGTTCACATGATCTTAAC

[0028] AAGATCACAGTCACAGAGACGATCAATCCAGATTTGAAGTTGGTACG

[0029] TGAGGATAGGCTTGACACTGATCTTTCCGTGATGAATGAAAAGATGCT

[0030] AAAAAACCATGTTGGGGATTCATCAAGTGAAAGTGCCGTGGCCGCTT

[0031] TGAGTATGAATAATGGTATGGCTGCTGATCTGAGAGCTGAAAATTTCA

[0032] GTCAGAGTTCACCCATAGACGAAAAAACTTTAGATATGGAAGCCAAT

[0033] TCCCCTATTACTGACTCTTCTTTGATATGGAATTTTCCATTGAACTTTG

[0034] GAAGTGGAGGAATCGAAGTCTGTAATCCTGAAAATGCAGTTGAGCCA

[0035] CTCAGAATAGTAGATGACAATGGCAGAATAGGTGGTGAAGTTGCTTC

[0036] TGCATCAGGGAGTGATTTTTGTGAAGCTGGCATGTCTTCTTCCCGAAG

[0037] GAAGGCCCGAGACGGGAAACAATGTAAAGTGGTGCAGACAAAAACG

[0038] AGTGCTCGCCATCTGAGAAAATCCTCTAGAAAGAAACAATCAGAAAG

[0039] AGACATTGAGTCAATTTTCAAGTGCTCAAAGCAGAAGAGGAGCTCTC

[0040] TTTTAAAAACAAGTCGTTCATCTGAGTGGGGATTACCAAGCAAGACC

[0041] ACTGAAATCTTCTTACAGAGCAATAATATTCCCTATGATGGACCTCCG

[0042] CATCATGAACCACAGAGATCTCAGGGAAATCTTAATAATGGAGAGCA

[0043] TAATAGAAGTTCTCATAACGGGAATGTAGAAGGATCTAACAGAAACA

[0044] TCCAAGCATCAAGTGGCTCTTGCCTTCGTTTGAAAGTTAAATTTGGTA

[0045] AATCAGGTGGCCAAAATCCTCTGAAC

[0046] ATTACAGTCTCTAAGGTCAGTGGTAACTCCTTACCTGGTAATGGTATT

[0047] GTAAAAGCAGGAACATGTTTAGAATTGCCAGGATCAGCACATTTTGG

[0048] TGAGGATAAAATGCAAACTGTGGAAACTAAAGAGGACTTGGTAGAGA

[0049] AAAGCAACCCTGTGGAGAAAGTTTCATATCTTCAGTCATCTGATTCTA

[0050] TGAGGGATAAGAAATATAACCAAGACGCTGGGGGTTTATGTAGGAAG

[0051] GTGGGTGGTGATGTTTTAGATGATGACCCACACCTTTCCTCTATTAGA

[0052] ATGGTTGAGGAGTGCGAGAGGGCCACTGGAACGCAGTCCCTGGACGC

[0053] TGAAACTTCACCAGATTCTGAAGTTATCAACTCTGTGCCTGATTCCAT

[0054] TGTCAATATTGAACATAAAGAGGGTCTGCATCATGGTTTTTTTAGCAC

[0055] TCCAGAAGATGTGGTCAAGAAGAACAGAGTATTAGAAAAAGAAGAT

[0056] GAATTGCGTGCTTCAAAATCTCCTTCAGAAAATGGTTCACATCTAATT

[0057] CCCAATGCAAAAAAAGCTAAACATCCGAAGTCTAAAAGTAATGGAAC

[0058] AAAGAAAGGTAAATCCAAGTTTTCCGAATCTGCCAAAGACGGGAGAA

[0059] AAAATGAATCACATGAAGGGGTAGAGCAACGCAAATCCCTGAATACC

[0060] AGTATGGGAAGGGATGATAGTGATTATCCTGAAGTAGGGAGAATAGA

[0061] GTCTCACAAAACAACAGGTGCCCTTCTAGATGCTGATATTGGGAAAA

[0062] CCAGTGCCACTTATGGTACCATATCATCAGATGTGACCCATGGGGAAA

[0063] TGGTTGTGGATGTTACTATTGAAGATAGCTATTCCACAGAGAGTGCCT

[0064] GGGTTCGATGTGATGATTGCTTTAAATGGCGACGAATACCTGCTTCTG

[0065] TTGTAGGATCAATTGACGAGAGCTCTAGATGGATCTGTATGAACAACT

[0066] CAGATAAAAGATTTGCGGATTGCTCAAAATCTCAAGAGATGTCAAAT

[0067] GAAGAAATTAATGAAGAGTTGGGCATAGGACAGGATGAAGCAGATG

[0068] CATATGATTGTGATGCGGCTAAAAGAGGGAAAGAAAAGGAACAGAA

[0069] GAGCAAGCGTTTGACAGGTAAGCAAAAGGCGTGCTTCAAGGCTATAA

[0070] AAACAAACCAGTTCCTTCATCGCAATCGTAAATCTCAAACAATTGACG

[0071] AGATAATGGTTTGTCACTGCAAACCATCACCTGATGGTAGGTTGGGTT

[0072] GTGGAGAAGAATGCCTCAATAGAATGCTTAACATTGAATGTCTTCAA

[0073] GGTACCTGTCCAGCTGGCGATTTGTGTTCAAATCAGCAGTTTCAAAAA

[0074] CGGAAGTATGTTAAGTTTGAGAGATTCCAATCCGGTAAGAAGGGTTA

[0075] TGGCCTGAGATTGCTCGAGGATGTACGAGAGGGGCAATTCCTAATTG

[0076] AATATGTTGGAGAGGTGCTTGATATGCAATCCTATGAGACTCGCCAAA

[0077] AAGAATATGCTTTCAAGGGTCAGAAACATTTCTATTTCATGACACTAA

[0078] ATGGGAATGAGGTAATAGATGCTGGTGCAAAGGGAAACCTAGGGCGT

[0079] TTCATTAACCATAGCTGTGAACCAAACTGCCGTACTGAAAAGTGGATG

[0080] GTGAATGGTGAAATTTGCGTTGGAATATTCTCCATGCAAGACCTTAAG

[0081] AAGGGTCAAGAGTTGACATTTGACTACAACTATGTGAGGGTTTTTGGT

[0082] GCTGCCGCCAAAAAGTGCTATTGTGGATCATCA

[0083] CATTGCAGAGGGTATATTGGGGGAGATCCTCTGAATGGTGATGTAATT

[0084] ATTCAAAGTGATTCAGATGAAGAGTATCCTGAACTTGTGATCCTTGAT

[0085] GATGATGAAAGTGGAGAAGGAATCTTAGGTGCAACATCTAGGACCTT

[0086] CACTGATGACGCTGACGAGCAAATGCCACAGAGCTTTGAAAAGGTTA

[0087] ATGGTTACAAGGACCTTGCTCCTGATAATACCCAAACACAGAGCTCA

[0088] GTATCTGTAAAACTTCCAGAGAGAGAAATTCCTCCACCTCTTCTTCAG

[0089] CCAACTGAAGTTTTGAAGGAACTTTCGTCAGGCATATCTATTACTGCT

[0090] GTCCAGCAAGAGGTTCCCGCTGAAAAGAAGACTAAAAGCACATCTCC

[0091] CACGTCCAGTTCTCTTAGCAGAATGTCCCCGGGTGGTACAAATTCTGA

[0092] TAAGACGACAAAGCATGGATCAGGAGAAGATAAAAAGATACTTCCAC

[0093] GTCCTCGTCCTCGTATGAAAACTTCTCGCTCATCTGAGTCTAGTAAGC

[0094] GAGACAAAGGAGGTATTTATCCTGGTGTTAACAAAGCACAGGTTATA

[0095] CCAGTTAATAAGTTGCAACAGCAGCCCATCAAATCTAAAGGATCAGA

[0096] GAAAGTTTCTCCCAGTATCGAAACATTTGAAGGGAAACTGAACGAGT

[0097] TACTAGATGCCGTAGGAGGGATAAGCAAGCGGAGGGATTCAGCAAAA

[0098] GGCTACTTGAAACTTCTGCTGCTCACTGCCGCTTCGCGAGGCACGGAT

[0099] GAAGAAGGAATTTATAGCAATCGAGATCTTTCAATGATTCTTGATGCC

[0100] CTTTTGAAGACAAAGTCAAAATCTGTTTTAGTGGACATAATCAACAAG

[0101] AATGGTCTGCAAATGTTACATAATATCATGAAACAATACCGGGGTGA

[0102] TTTTAAAAGGATCCCTATAATCCGGAAACTTCTGAAGGTATTAGAGTA

[0103] TCTTGCCACAAGGAAAATCCTTGCACTGGAGCATATAATCAGAAGGC

[0104] CTCCTTTTGCAGGGATGGAAAGCTTTAAGGACTCTGTTCTATCATTCA

[0105] CCGAGCATGACGACTATACGGTTCATAATATTGCGCGGAGCTTCCGAG

[0106] ACAGATGGATACCTAAGCATTTTAGAAAACCTTGGCGCATCAACAGG

[0107] GAGGAGAGATCGGAGTCTATGAGGTCACCTATAAACAGAAGGTTCAG

[0108] AGCATCACAAGAACCTCGATATGATCATCAGTCCCCAAGACCTGCAG

[0109] AACCAGCTGCGTCTGTCACATCATCAAAGGCTGCAACTCCTGAAACA

[0110] GCATCTGTATCTGAGGGATATTCAGAACCTAATTCCGGTCTTCCTGAG

[0111] ACAAATGGACGCAAGCGCAAAAGCAGATGGGACCAGCCGTCTAAGA

[0112] CAAAAGAACAAAGAATCATGACTATCTTGTCCCAACAAACAGATGAA

[0113] ACTAATGGAAATCAGGATGTCCAAGATGACCTTCCACCCGGGTTTTCA

[0114] TCACCCTGCACAGATGTGCCTGATGCAATTACTGCACAGCCGCAACAA

[0115] AAGTTCCTTTCTCGCTTACCAGTTTCCTATGGGATTCCACTTAGCATTG

[0116] TTCATCAATTTGGTTCACCTGGCAAAGAGGACCCGACCACCTGGTCTG

[0117] TTGCTCCTGGCATGCCGTTCTATCCATTTCCACCTCTACCGCCAGTGTC

[0118] TCATGGTGAGTTTTTTGCTAAGAGAAACGTAAGAGCCTGTTCCTCGTC

[0119] CATGGGAAACCTGACTTACTCCAATGAGATCTTACCAGCTACACCTGT

[0120] GACTGACTCGACCGCTCCAACCCGTAAGAGAGAGTTGTTTTCATCTGA

[0121] TATAGGGACAACTTACTTTCGGCAACAGAAACAGAGTGTTCCTCCATG

[0122] GTTGCGGAACAATGGGGGGGAGAAAACAGCAAACAGCCCTATACCTGGAAATCTAACTTTAGAGAAGAAGCTCAACAGTTAA(SEQ ID NO.1);

[0123] MDCKENGVGDASGCNIDANSLASNLAMNTNEDFYEKLSSRGQNLDSVSSLEIPQTASSVNHTIEGQRKCFTEIEQMGYGNSNSQEDAGNTDDDLYVCYNADDTQEQGVVSGELEQSQELICDTDLLVNCNKLDDGKESQDTNVSLVSIFSGSMQEKEAPQAKEDEGYGGTTLPIGGSGIDTESTFVNDAPEQFESLETTKHIKPDEVESDGISYRFDDGGKEGRNGPSSDLDTGSSDDISLSQSFSFPDSLLDSSVFGCSATESYLEDAIDIEGNGTIVVSPSLAITEMLNNDDGGLCSHDLNKITVTETINPDLKLVREDRLDTDLSVMNEKMLKNHVGDSSSESAVAALSMNNGMAADLRAENFSQSSPIDEKTLDMEANSPITDSSLIWNFPLNFGSGGIEVCNPENAVEPLRIVDDNGRIGGEVASASGSDFCEAGMSSSRRKARDGKQCKVVQTKTSARHLRKSSRKKQSERDIESIFKCSKQKRSSLLKTSRSSEWGLPSKTTEIFLQSNNIPYDGPPHHEPQRSQGNLNNGEHNRSSHNGNVEGSNRNIQASSGSCLRLKVKFGKSGGQNPLNITVSKVSGNSLPGNGIVK

[0124] AG

[0125] TCLELPGSAHFGEDKMQTVETKEDLVEKSNPVEKVSYLQSSDSMRDKKY

[0126] NQDAGGLCRKVGGDVLDDDPHLSSIRMVEECERATGTQSLDAETSPDSE

[0127] VINSVPDSIVNIEHKEGLHHGFFSTPEDVVKKNRVLEKEDELRASKSPSEN

[0128] GSHLIPNAKKAKHPKSKSNGTKKGKSKFSESAKDGRKNESHEGVEQRKS

[0129] LNTSMGRDDSDYPEVGRIESHKTTGALLDADIGKTSATYGTISSDVTHGE

[0130] MVVDVTIEDSYSTESAWVRCDDCFKWRRIPASVVGSIDESSRWICMNNS

[0131] DKRFADCSKSQEMSNEEINEELGIGQDEADAYDCDAAKRGKEKEQKSKR

[0132] LTGKQKACFKAIKTNQFLHRNRKSQTIDEIMVCHCKPSPDGRLGCGEECL

[0133] NRMLNIECLQGTCPAGDLCSNQQFQKRKYVKFERFQSGKKGYGLRLLED

[0134] VREGQFLIEYVGEVLDMQSYETRQKEYAFKGQKHFYFMTLNGNEVIDAG

[0135] AKGNLGRFINHSCEPNCRTEKWMVNGEICVGIFSMQDLKKGQELTFDYN

[0136] YVRVFGAAAKKCYCGSSHCRGYIGGDPLNGDVIIQSDSDEEYPELVILDD

[0137] DESGEGILGATSRTFTDDADEQMPQSFEKVNGYKDLAPDNTQTQSSVSV

[0138] KLPEREIPPPLLQPTEVLKELSSGISITAVQQEVPAEKKTKSTSPTSSSLSRM

[0139] SPGGTNSDKTTKHGSGEDKKILPRPRPRMKTSRSSESSKRDKGGIYPGVN

[0140] KAQVIPVNKLQQQPIKSKGSEKVSPSIETFEGKLNELLDAVGGISKRRDSA

[0141] KGYLKLLLLTAASRGTDEEGIYSNRDLSMILDALLKTKSKSVLVDIINKN

[0142] GLQMLHNIMKQYRGDFKRIPIIRKLLKVLEYLATRKILALEHIIRRPPFAG

[0143] MES

[0144] FKDSVLSFTEHDDYTVHNIARSFRDRWIPKHFRKPWRINREERSESMRSPI

[0145] NRRFRASQEPRYDHQSPRPAEPAASVTSSKAATPETASVSEGYSEPNSGLP

[0146] ETNGRKRKSRWDQPSKTKEQRIMTILSQQTDETNGNQDVQDDLPPGFSSP

[0147] CTDVPDAITAQPQQKFLSRLPVSYGIPLSIVHQFGSPGKEDPTTWSVAPGM

[0148] PFYPFPPLPPVSHGEFFAKRNVRACSSSMGNLTYSNEILPATPVTDSTAPT

[0149] RKRELFSSDIGTTYFRQQKQSVPPWLRNNGGEKTANSPIPGNLTLEKKLNS(SEQ ID NO.2).

[0150] After knocking out the methyltransferase gene EFS in Arabidopsis thaliana, an EFS gene knockout mutant was obtained (this mutant was purchased from the TAIR Arabidopsis thaliana database, and the mutant number is Salk_065480). The sterilized seeds of the Arabidopsis thaliana EFS gene knockout mutant were evenly spread on 1 / 2 MS solid medium. After 3 days of dark treatment at 4°C, the plates were placed vertically under the growth condition of long-day (16 hours of light, 8 hours of darkness) at 21°C for 3 days. Subsequently, the seedlings were transferred to 1 / 2 MS solid medium containing 100 mM NaCl and 125 mM NaCl and blank 1 / 2 MS solid medium with toothpicks, and were placed under the growth condition of long-day at 21°C. After 7 - 10 days of vertical growth, the root lengths were measured and photographed. The specific results are shown in Figure 1 .

[0151] It can be seen from Figure 1 that under normal conditions, that is, on the blank 1 / 2 MS solid medium, there was no obvious change in the root lengths of the wild-type Col and the mutant efs. After treatment with 100 mM NaCl and 125 mM NaCl, the growth of the root length of the mutant efs was significantly inhibited.

[0152] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. Use of methyltransferase gene EFS in regulating plant salt stress resistance, wherein, The nucleotide sequence of the methyltransferase gene EFS is shown in SEQ ID NO.

1.

2. The application according to claim 1, wherein The plant is Arabidopsis thaliana.

3. The application according to claim 1, wherein Knocking out the methyltransferase gene EFS reduces the salt stress resistance performance of the plant.

4. A preparation for regulating plant resistance to salt stress, characterized in that, It includes the methyltransferase gene EFS described in claim 1 or the protein encoded by this gene.

5. A method for regulating plant resistance to salt stress, characterized in that, It includes the following steps: Regulating the salt stress resistance performance of plants by knocking out or overexpressing the methyltransferase gene EFS.

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