CN114437192B - Tomato leaf miner chromatin remodelling factor ISWI, and encoding gene and application thereof - Google Patents

Abstract

The application relates to the field of insect control, in particular to a tomato leaf miner chromatin remodelling factor ISWI, and a coding gene and application thereof. The amino acid sequence of the chromatin remodeling factor ISWI is shown in SEQ ID NO: 2. The application clones and obtains the chromatin remodelling factor ISWI gene from the new important invasive pest tomato leaf miner for the first time, and the invasive pest ovum injection of the gene dsRNA leads to the ovarian malformation, the remarkable reduction of spawning quantity and the remarkable shortening of the adult life. The obtained result provides a new way for preventing and controlling new invasion pests by utilizing RNAi.

Description

A tomato leafminer chromatin remodeling factor ISWI and its encoding gene and application

Technical field

The invention relates to the field of insect control, and specifically relates to a tomato leafminer chromatin remodeling factor ISWI and its encoding gene and application.

Background technique

The tomato leafminer Tuta absoluta is native to Peru, South America. It is a quarantine pest in the world and causes devastating disasters to tomatoes and other crops. Tomato leafminer mainly causes damage as larvae. Female adults lay eggs on newly unfolded leaves. Once the larvae hatch, they sneak into the tissue of the host plant and feed on the leaves. If control measures are not taken, this pest can reduce tomato yields by 80-100%. At the same time, with the continued use of chemical pesticides, their resistance to pesticides continues to increase, and they can quickly develop resistance to various pharmaceutical targets.

The phenomenon of RNA interference (RNAi) is an evolutionarily conserved defense mechanism against invasion by transgenics or foreign viruses. RNAi exists widely in the biological world. RNAi technology can silence certain genes in insects, thereby enhancing or losing certain abilities of insects. It can also inhibit the expression of functional genes at a specific time to stop the development of insects at a certain stage. Because RNA interference technology is insect-selective and gene-specific, it is not only an important method for studying gene function, but also the most likely bioengineering technology to be applied to pest control so far. One of the key steps in using RNAi technology for biological pest control is to screen target genes.

Chromatin remodeling is one of the important components of epigenetics. There are a series of specific chromatin remodeling factors in eukaryotic cells that can promote the dynamic combination of DNA and proteins in chromatin. These factors are multi-subunit complexes that use the energy generated by ATP hydrolysis to change the state of chromatin by sliding nucleosomes, thereby regulating the expression of specific genes. Chromatin remodeling plays a key role in the expression regulation of genes related to development and reproduction of organisms. The chromatin remodeling factor ISWI (imitation SWI) can participate in the regulation of development and reproduction of organisms and plays an important role in regulating the expression of genes related to development and reproduction. ISWI silence results in shortened lifespan, ovarian malformation, and reduced egg production. Therefore, the present invention provides a new molecular target for pest control based on RNA interference technology, and is expected to provide new ideas for the prevention and control of tomato leafminer and achieve effective prevention and control effects.

Contents of the invention

The purpose of the present invention is to provide a tomato leafminer chromatin remodeling factor.

Another object of the present invention is to provide a gene encoding the tomato leafminer chromatin remodeling factor ISWI.

Another object of the present invention is to provide applications of the above-mentioned encoding genes.

According to the specific embodiment of the present invention, the tomato leafminer chromatin remodeling factor ISWI was cloned for the first time, and its full-length cDNA nucleotide sequence is shown in SEQ ID NO: 1:

ATGTCACAACCTGATGAAGCTATGGATGTAGGGGACGTGGAAGAAAACTCTAATGAGTCGTCCAGCGATACCACGTCCTCAAAGGGTAAAGAAGGAGACTTCGAAAGTAAAATTGAGACCGATCGTTCCAAGCGTTTTGATTTTTTGTTGAAACAAACTGAAATATTCTCGCATTTCATGACCAATGCGCCTAAAACCAGTAGTCCTCCTAAGGCTAAAGCCGGTCGACCTAAAAAGGTGAAAGAAGATCTGCCTGAGCAGTC TGAGGATGCTTCTGCAGCAGACCATCGGCACCGCAAGACTGAACAGGAAGAAGATGAGGAACTCCTTGCTGAGACAAACGCTAAGATTAAGCCAATATTTCGCTTTGAAGCATCACCTCCTTACATCAAGAATGGAGAAATGAGAGATTATCAAGTGAGAGGTCTCAACTGGATGATATCTCTCTATGAGAATGGTATTAATGGTATCTTGGCTGATGAGATGGGTCTGGGTAAAACATTACAAACCATTTCTCTTCTTGGC TACATGAAAAATTTCAGAAATATCCCTGGTCCACACATAGTAATTGTGCCAAAATCAACCCTAACCAACTGGATGAATGAGTTCAAAAAATGGTGTCCTTCCCTAAGAGCAGTGTGTCTTATTGGAGATCAGGAAACAAGGAACACATTCATCCGTGAAGTACTGATGCCTGGCAACTGGGATGTGTGCATCACCTCTTACGAGATGATTATCCGTGAGAGATCAGTATTTAAGAAGTTCACTTGGCGCTACATGGTGATTGACGA AGCCCATCGAATCAAGAATGAGAAGTCAAAGTTGTCAGAGCTTCTCCGGGAGTTCAAGAGTATGAACCGGCTGCTGCTCACTGGCACACCGTTGCAAAACAATCTTCACGAGCTGTGGGCACTTCTTAACTTCTTGTTACCAGATGTATTCAACAGCTCTGATGATTTTGACTCTTGGTTCAACCAATGCAGCTCTCGGTGATAATCAGCTGGTGTCCCGGTTGCATGCTGTGTTGAGACCGTTCTTACTCAGACG TCTCAAATCTGAAGTAGAAAAGAAATTGAAACCCAAAAAGGAACTCAAAGTTTATGTTGGATTGAGTAAGATGCAAAGGGAGTGGTATACTAAAGTACTCATGAAGGATATTGATATTGTAAACGGCGCGGGCAAAGTCGAAAAGATGCGTCTCCAGAACATCCTGATGCAGCTCCGCAAGTGCTGCAACCACCCGTACCTGTTCGACGGCGCGGAGCCCGGCCCGCCCTACACCACGGACGAGCATCTCGTCTACAACTGCGGGA AACTCGCCATACTCGACAAACTGCTGCCCAAGCTGCAGCAGCAGGACTCCAGGGTGCTCATCTTCTCGCAGATGACCAGGATGCTGGATATTCTGGAGGATTATTGCCTGTGGAGGCAGTACAAGTACTGCCGCTTAGACGGTCAGACCCCACATGAAGACCGCAACCGACAGATCGAGGAGTATAACGCGGAAGGCTCCGAGAAGTTCGTGTTCATGTTGTCCACCCGCGCCGGCGGGCTGGGGATCAACCTCACCTCGG CAGATGTCGTCATCATATACGACTCCGACTGGAACCCGCAGATGGACCTGCAGGCGATGGACCGCGCGCATCGTATTGGGCATGAAACAAGTGCGCGTATTCAGACTGATCACGGAGAACACGGTGGAAGAGAAGATAGTGGAGCGCGCGGAAGTGAAACTACGCCTAGACAAGCTCGTAATCCAGTCCGGCCGGCTCGTGGACCAGAAGAACCAGCTCAACAAGGACGAAATGCTCAACATGATCAGGCACGGGGGCCAACC ACGTCTTCTCCTCGAAGGACTCCGAGATCACCGATGAGGATATTGACTCTATTCTAGCCAAAGGAGAGACCAAGACAGAAGAATTGAAACAGAAGTTGGAGAGCCTGGAGGAGTCATCTCTGAGAGCGTTCTCGATGGACACCCCCGGCGCGACCGACTCCGTCTACCAATTCGAAGGCGAGGACTACCGGGAGAAGCAGAAGTCGCACCCGCTGGGCAGCTGGATCGAGCCGCCGAAGCGCGAGCGCAAGGCCA ACTACGCGGTGGACGCGTACTTCCGCGAGGCTGCGCGTCTCCGAGCCCAAGCGCCCGAAAGTGCAGGCGCCGCGCCCCCCCAAGCAGCCCATCGTGCAGGACTTCCAGTTCTTCCCGCCGAGGCTGTTCGAGCTGCTGGACCAGGAGATCTACCACTACAGAAAGACATTGGGGTACAAAGTGCCGCGTAACCCGGAGCTGGGCCCCGACGCGGCCAAGATACAGAGAGAGGAGCAGAGGAAGATCGACGACGCCGAA GCGCTCACCGAGGAGGAGGTTGCTGAGAAAGAAAACCTTCTCACTCAGGGTTTCACAAACTGGACAAAACGCGACTTCAACCAGTTCATCAAAGCTAATGAGAAGTATGGAAGGGATGACATCGAAAATATTGCAAAAGATGTCGAAGGAAAAACGCCCGAAGAGGTGATGGAATACTCGGCAGTGTTCTGGGAGCGCTGCCACGAGCTGCAGGACGTGGACCGGATCATGGGGCAGATCGAGCGCGGCGAGGCCAAGATACAGC GCCGCGCCTCCATCAAGAAGGCGCTCGACGCCAAGATGGCGCGCTACCGGGCCGCCCTTCCACCAGCTCAGGATATCCTACGGGACTAACAAGGGGAAGAACTATGTCGAAGAAGAAGACAGGTTCCTGGTGTGCATGCTGCACAAGCTGGGCTTCGACAAGGAGAACGTGTACGAGGAGCTGCGCGCCGCCGTGCACGCCGCGCCGCAGTTCCGCTTCGACTGGTTCCTCAAGTCGCGCACGGCCGTGGAGCT GCAGCGCAGATGCAACACATTGATCACACTCATCGAAAGAGAGAACCAAGAGCTCGAAGAGAAGGAGCGCGCCGAGAAGAAAAAGAAGAGCGGCAGCGCCAACCAGAACACGCCCGGCGCCGCCAGCAAGGGCGCCGGCGCCGGCAAGCGCAAGGCAGACGCCGCGCAGGACTCCGCGCAGAAACAGAAGAAGAAGAAGAAATGA

The amino acid sequence of tomato leafminer chromatin remodeling factor ISW is shown in SEQ ID NO:2:

MSQPDEAMDVGDVEENSNESSSDTTSSKGKEGDFESKIETDRSKRFDFLLKQTEIFSHFMTNAPKTSSPPKAKAGRPKKVKEDLPEQSEDASAADHRHRKTEQEEDEELLAETNAKIKPIFRFEASPPYIKNGEMRDYQVRGLNWMISLYENGINGILADEMGLGKTLQTISLLGYMKNFRNIPGPHIVIVPKSTLTNWMNEFKKWCPSLRAVCLIGDQ ETRNTFIREVLMPGNWDVCITSYEMIIRERSVFKKFTWRYMVIDEAHRIKNEKSKLSELLREFKSMNRLLLTGTPLQNNLHELWALLNFLLPDVFNSSDDFDSWFNTNAALGDNQLVSRLHAVLRPFLLRRLKSEVEKKLKPKKELKVYVGLSKMQREWYTKVLMKDIDIVNGAGKVEKMRLQNILMQLRKCCNHPYLFDGAEPGPPYTTDEHLVY NCGKLAILDKLLPKLQQQDSRVLIFSQMTRMLDILEDYCLWRQYKYCRLDGQTPHEDRNRQIEEYNAEGSEKFVFMLSTRAGGLGINLTSADVVIIYDSDWNPQMDLQAMDRAHRIGQMKQVRVFRLITENTVEEKIVERAEVKLRLDKLVIQSGRLVDQKNQLNKDEMLNMIRHGANHVFSSKDSEITDEDIDSILAKGETKTE ELKQKLESLEESSLRAFSMDTPGATDSVYQFEGEDYREKQKSHPLGSWIEPPKRERKANYAVDAYFREALRVSEPKAPKVQAPRPPKQPIVQDFQFFPPRLFELLDQEIYHYRKTLGYKVPRNPELGPDAAKIQREEQRKIDDAEALTEEEVAEKENLLTQGFTNWTKRDFNQFIKANEKYGRDDIENIAKDVEGKTPEEVMEYSAVFWERCHE LQDVDRIMGQIERGEAKIQRRASIKKALDAKMARYRAPFHQLRISYGTNKGKNYVEEEDRFLVCMLHKLGFDKENVYEELRAAVHAAPQFRFDWFLKSRTAVELQRRCNTLITLIERENQELEEKERAEKKKKSGSANQNTPGAASKGAGAGKRKADAAQDSAQKQKKKKK

The above amino acid sequence has the typical structural characteristics of ISWI protein, including N-terminal ATPase domain (DEXDc and HELICc) and C-terminal HAND-SANT-SLIDE domain.

The present invention provides the application of the above tomato leafminer chromatin remodeling factor ISWI. RNAi was performed on tomato leafminers, and the results showed that the ovaries of tomato leafminers that fed on dsISWI developed malformations and the number of eggs laid by adults was significantly reduced, indicating that the ISWI gene plays a key role in the reproductive development of tomato leafminers. The dsRNA synthesized based on the tomato leafminer ISWI gene fragment of the present invention promotes ovarian malformation, significantly reduces adult egg laying, and significantly shortens the adult lifespan. It can be used as a molecular target for RNAi control of tomato leafminer and has the ability to prevent and control this invasive pest. important practical significance.

For the first time, the present invention cloned the chromatin remodeling factor ISWI gene from the newly emerging important invasive pest tomato leafminer. Injection of gene dsRNA into the eggs of the above-mentioned invasive pest resulted in ovarian malformation, significant reduction in egg production, and significantly shortened adult lifespan. The results obtained provide new ways to use RNAi to control emerging invasive pests.

Description of the drawings

Figure 1 shows the changes in the ovary of female adults under the condition of injection of ISWI gene dsRNA and dsEGFP;

Figure 2 Changes in ISWI gene expression profiles at different developmental stages;

Figure 3 shows the changes in the expression of ISWI under the conditions of injection of ISWI gene dsRNA and dsEGFP;

Figure 4 shows the changes in the longevity of tomato leafminer adults treated with ISWI gene dsRNA injection;

Figure 5 shows the changes in the number of eggs laid by tomato leafminer adults treated with ISWI gene dsRNA;

Figure 6 shows the changes in the hatching rate of tomato leafminer eggs treated with ISWI gene dsRNA injection.

Detailed ways

Example 1: Cloning of full-length cDNA sequence of ISWI gene of tomato leafminer

Take 4 tomato leafminer larvae and put them into a 1.5 mL centrifuge tube. After freezing in liquid nitrogen, grind them into powder using a grinding rod. Then extract the RNA and store it at -80°C for later use. According to the instructions of TransScript All-in-OneFirst-Strand cDNA Synthesis SuperMix for PCR, the extracted RNA was reverse transcribed to synthesize cDNA. Use cDNA as a template to design primers and perform PCR amplification. Design primers as shown in Table 1:

Table 1 Primer sequences for cloning the full-length cDNA of TaISWI gene

Using the primers in Table 1, the full-length cDNA sequence of the ISWI gene was obtained through PCR amplification, which is 3078 bp. The obtained gene has a nucleotide sequence as shown in SEQ ID NO: 1, and the gene encodes 1025 nucleotides as shown in SEQ ID NO: The amino acid sequence shown in 2. The conserved domain analysis of the amino acid sequence encoded by the cloned gene showed that it has the typical structural characteristics of ISWI proteins: it contains N-terminal ATPase domain (DEXDc and HELICc) and C-terminal HAND-SANT-SLIDE domain.

Example 2: Analysis of the effect of ISWI gene on reproductive development of tomato leafminer

1. Synthesis of dsRNA

(1) Design and synthesize the primer sequence with T7 promoter:

T7+dsISWI-F：5’-TAATACGACTCACTATAGGGTATGGATGTAGGGGACG-3’

T7+dsISWI-R: 5’-TAATACGACTCACTATAGGGCCAGTTGGTTAGGGTTG-3’.

Synthesized by Shanghai Sangon Bioengineering Technology Services Co., Ltd.

(2) Total RNA extraction and cDNA synthesis: Same as Example 1.

(3) T7 primer PCR amplification and product purification. The purified PCR product is the template for synthesizing dsRNA. Use the kit to synthesize and purify dsRNA, and follow the instructions of the kit.

2. dsRNA injection method

(1) Collect the mature female pupae of the tomato leafminer before they emerge, arrange them and adhere them to the surface of a coverslip that has been previously affixed with breathable double-sided tape. Using a Femtojet Express (Eppendorf) microinjector, dsISWI (injection volume: 0.6 μg dsRNA), dsEGFP and ddH ₂ O (0.1 μl) were injected into the internodes. After injection, place the female pupa single-headed in a breathable 1.5mL centrifuge tube. Once it emerges, pair it with the newly emerged wild-type male adult and place it in a plastic box alone, and provide enough fresh tomato leaves for it to feed. The number of eggs laid and lifespan of adult worms were counted. At the same time, adult worms that had emerged for 4 days were randomly selected, their ovaries were dissected, and their morphological characteristics were observed. In order to detect the efficiency of RNAi interference, 4 tomato leafminers 72 hours after injection were used to detect expression levels, and 4 biological replicates were set for each treatment. As shown in Figure 1, it shows the changes in female adult ovaries under the conditions of injection of ISWI gene dsRNA and dsEGFP. Compared with the control treatment, the number of mature eggs in the female adult ovaries after injection of ISWI gene dsRNA was significantly reduced.

(2) The relative expression of genes was calculated by the 2 ^-ΔΔCT method. The results are shown in Figure 2, which shows the changes in ISWI gene expression profiles at different developmental stages. The ISWI gene has the highest expression in the two developmental stages of egg and pupa. Figure 3 shows the changes in ISWI expression under the conditions of injection of ISWI gene dsRNA and dsEGFP. The results show that injection of dsISWI dsRNA can significantly knock down the expression of ISWI gene.

SAS 9.4 statistical software was used to analyze the egg laying volume (Figure 5) and female life span (Figure 4) of tomato leafminer adults after injection of different solutions. The results showed that the total egg laying volume of tomato leafminer adults fed dsISWI gene dsRNA and life span were significantly lower than those of the dsEGFP injection group (P<0.05). Figure 6 shows the change in hatching rate of tomato leafminer eggs after injection of ISWI gene dsRNA. Compared with the control, injection of dsISWI can significantly inhibit the hatching of tomato leafminer eggs.

At the same time, BLAST at NCBI (http://blast.ncbi.nlm.nih.gov/) shows that the injected target sequence fragment is a sequence unique to the ISWI gene, thus ensuring that the interference effect is for the ISWI gene of tomato leafminer Therefore, it shows that the ISWI gene of the present invention plays a key role in the development and reproduction of tomato leafminer.

The present invention clones the full-length cDNA of the ISWI gene from the tomato leafminer. Fluorescence quantitative PCR shows that the expression level of the ISWI gene is significantly reduced after injecting double-stranded RNA of the target gene; finally, by injecting the ISWI gene dsRNA, the female tomato leafminer is The number of eggs laid and lifespan of insects are significantly reduced. According to the specific embodiment of the present invention, the test results clarify that the ISWI gene plays a key role in the development and reproduction of the South American tomato leafminer. The present invention lays a foundation for the research on effective prevention and control of tomato leafminer using RNAi, and provides a methodological basis for reducing the damage of tomato leafminer through related research on chromatin remodeling factors in the future.

The above embodiments are only used to explain the technical solutions of the present application and do not limit the protection scope of the present application.

sequence list

<110> Institute of Plant Protection, Chinese Academy of Agricultural Sciences

<120> A tomato leafminer chromatin remodeling factor ISWI and its encoding gene and application

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<170> SIPOSequenceListing 1.0

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<213> Tomato leafminer (Tuta absoluta)

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<212> PRT

<213> Tomato leafminer (Tuta absoluta)

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Asp Phe Glu Ser Lys Ile Glu Thr Asp Arg Ser Lys Arg Phe Asp Phe

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Leu Leu Lys Gln Thr Glu Ile Phe Ser His Phe Met Thr Asn Ala Pro

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Lys Ser Thr Leu Thr Asn Trp Met Asn Glu Phe Lys Lys Trp Cys Pro

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Ser Leu Arg Ala Val Cys Leu Ile Gly Asp Gln Glu Thr Arg Asn Thr

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Phe Ile Arg Glu Val Leu Met Pro Gly Asn Trp Asp Val Cys Ile Thr

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Ser Tyr Glu Met Ile Ile Arg Glu Arg Ser Val Phe Lys Lys Phe Thr

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Trp Arg Tyr Met Val Ile Asp Glu Ala His Arg Ile Lys Asn Glu Lys

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Ser Lys Leu Ser Glu Leu Leu Arg Glu Phe Lys Ser Met Asn Arg Leu

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Leu Leu Thr Gly Thr Pro Leu Gln Asn Asn Leu His Glu Leu Trp Ala

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Leu Leu Asn Phe Leu Leu Pro Asp Val Phe Asn Ser Ser Asp Asp Phe

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Asp Ser Trp Phe Asn Thr Asn Ala Ala Leu Gly Asp Asn Gln Leu Val

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Ser Arg Leu His Ala Val Leu Arg Pro Phe Leu Leu Arg Arg Leu Lys

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Ser Glu Val Glu Lys Lys Leu Lys Pro Lys Lys Glu Leu Lys Val Tyr

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Val Gly Leu Ser Lys Met Gln Arg Glu Trp Tyr Thr Lys Val Leu Met

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Lys Asp Ile Asp Ile Val Asn Gly Ala Gly Lys Val Glu Lys Met Arg

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Leu Gln Asn Ile Leu Met Gln Leu Arg Lys Cys Cys Asn His Pro Tyr

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Leu Phe Asp Gly Ala Glu Pro Gly Pro Pro Tyr Thr Thr Asp Glu His

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Leu Val Tyr Asn Cys Gly Lys Leu Ala Ile Leu Asp Lys Leu Leu Pro

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Lys Leu Gln Gln Gln Asp Ser Arg Val Leu Ile Phe Ser Gln Met Thr

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Arg Met Leu Asp Ile Leu Glu Asp Tyr Cys Leu Trp Arg Gln Tyr Lys

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Tyr Cys Arg Leu Asp Gly Gln Thr Pro His Glu Asp Arg Asn Arg Gln

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Ile Glu Glu Tyr Asn Ala Glu Gly Ser Glu Lys Phe Val Phe Met Leu

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Ser Thr Arg Ala Gly Gly Leu Gly Ile Asn Leu Thr Ser Ala Asp Val

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Val Ile Ile Tyr Asp Ser Asp Trp Asn Pro Gln Met Asp Leu Gln Ala

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Met Asp Arg Ala His Arg Ile Gly Gln Met Lys Gln Val Arg Val Phe

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Val Asp Gln Lys Asn Gln Leu Asn Lys Asp Glu Met Leu Asn Met Ile

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Arg His Gly Ala Asn His Val Phe Ser Ser Lys Asp Ser Glu Ile Thr

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Asp Glu Asp Ile Asp Ser Ile Leu Ala Lys Gly Glu Thr Lys Thr Glu

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Glu Leu Lys Gln Lys Leu Glu Ser Leu Glu Glu Ser Ser Leu Arg Ala

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Phe Ser Met Asp Thr Pro Gly Ala Thr Asp Ser Val Tyr Gln Phe Glu

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Gly Glu Asp Tyr Arg Glu Lys Gln Lys Ser His Pro Leu Gly Ser Trp

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Ile Glu Pro Pro Lys Arg Glu Arg Lys Ala Asn Tyr Ala Val Asp Ala

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Tyr Phe Arg Glu Ala Leu Arg Val Ser Glu Pro Lys Ala Pro Lys Val

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Gln Ala Pro Arg Pro Pro Lys Gln Pro Ile Val Gln Asp Phe Gln Phe

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Phe Pro Pro Arg Leu Phe Glu Leu Leu Asp Gln Glu Ile Tyr His Tyr

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Arg Lys Thr Leu Gly Tyr Lys Val Pro Arg Asn Pro Glu Leu Gly Pro

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Asp Ala Ala Lys Ile Gln Arg Glu Glu Gln Arg Lys Ile Asp Asp Ala

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Glu Ala Leu Thr Glu Glu Glu Val Ala Glu Lys Glu Asn Leu Leu Thr

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Gln Gly Phe Thr Asn Trp Thr Lys Arg Asp Phe Asn Gln Phe Ile Lys

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Ala Asn Glu Lys Tyr Gly Arg Asp Asp Ile Glu Asn Ile Ala Lys Asp

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Val Glu Gly Lys Thr Pro Glu Glu Val Met Glu Tyr Ser Ala Val Phe

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Trp Glu Arg Cys His Glu Leu Gln Asp Val Asp Arg Ile Met Gly Gln

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Ile Glu Arg Gly Glu Ala Lys Ile Gln Arg Arg Ala Ser Ile Lys Lys

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Ala Leu Asp Ala Lys Met Ala Arg Tyr Arg Ala Pro Phe His Gln Leu

885 890 895

Arg Ile Ser Tyr Gly Thr Asn Lys Gly Lys Asn Tyr Val Glu Glu Glu

900 905 910

Asp Arg Phe Leu Val Cys Met Leu His Lys Leu Gly Phe Asp Lys Glu

915 920 925

Asn Val Tyr Glu Glu Leu Arg Ala Ala Val His Ala Ala Pro Gln Phe

930 935 940

Arg Phe Asp Trp Phe Leu Lys Ser Arg Thr Ala Val Glu Leu Gln Arg

945 950 955 960

Arg Cys Asn Thr Leu Ile Thr Leu Ile Glu Arg Glu Asn Gln Glu Leu

965 970 975

Glu Glu Lys Glu Arg Ala Glu Lys Lys Lys Lys Ser Gly Ser Ala Asn

980 985 990

Gln Asn Thr Pro Gly Ala Ala Ser Lys Gly Ala Gly Ala Gly Lys Arg

995 1000 1005

Lys Ala Asp Ala Ala Gln Asp Ser Ala Gln Lys Gln Lys Lys Lys Lys

1010 1015 1020

Lys

1025

Claims (6)

1. Tomato leafminer chromatin remodeling factor ISWI , characterized in that its amino acid sequence is shown in SEQ ID NO: 2. 2. Tomato leafminer chromatin remodeling factor gene, characterized in that the gene encodes the tomato leafminer chromatin remodeling factor ISWI according to claim 1. 3. The tomato leafminer chromatin remodeling factor gene according to claim 2, characterized in that the nucleotide sequence of the tomato leafminer chromatin remodeling factor gene is shown in SEQ ID NO: 1. 4. A recombinant expression vector comprising the tomato leafminer chromatin remodeling factor gene according to claim 2. 5. A recombinant strain comprising the tomato leafminer chromatin remodeling factor gene according to claim 2. 6. Application of the tomato leafminer chromatin remodeling factor gene of claim 2 as an RNAi interference target that interferes with the following traits, by feeding the tomato leafminer chromatin of claim 2 dSRNA fragment of the remodeling factor gene, (1) Reduce the number of eggs laid by female adults of tomato leafminer; (2) Shorten the lifespan of female adults of tomato leafminer, The dSRNA fragment of the tomato leafminer chromatin remodeling factor gene was obtained by the following primers: T7+ds ISWI -F：5'-TAATACGACTCACTATAGGGTATGGATGTAGGGGACG-3' T7+ds ISWI -R: 5'-TAATACGACTCACTATAGGGCCAGTTGGTTAGGGTTG-3'.