CN109575112B - Application of Phyllostachys pubescens PhJAZ1 gene in pest resistance difference of bamboo leaves in large and small years - Google Patents
Application of Phyllostachys pubescens PhJAZ1 gene in pest resistance difference of bamboo leaves in large and small years Download PDFInfo
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Abstract
The invention discloses a moso bambooPhJAZThe application of 1 gene in the difference of the insect resistance of the bamboo leaves in the year and the year belongs to the field of biotechnology. The invention clones to obtain the moso bambooPhThe gene sequence of the JAZ1 gene is shown as SEQ ID No.1, and the amino acid sequence thereof is shown as SEQ ID No. 2. Construction ofPhOverexpression vector of JAZ1 gene, and transformation of Arabidopsis thaliana by overexpression vectorjaz1Mutant to obtain Phyllostachys pubescensPhJAZ1And (3) gene overexpression plants.PhJAZ1The expression level of the gene in the bamboo of the old is obviously higher than that of the gene in the bamboo of the young. Through moso bambooPhJAZ1The gene overexpression inhibits the synthesis of tannin as an insect-resistant secondary metabolite in plants, and the characteristic is utilized to be beneficial to controlling the major outbreak of moso bamboo leaf pests, so that the gene overexpression has important significance on reasonable management and management of moso bamboo forests.
Description
Technical Field
The invention belongs to the field of biotechnology, and particularly relates to a biological enzyme inhibitorRelates to a moso bambooPhJAZ1The application of the gene in the difference of insect resistance of bamboo leaves in the year of old and young.
Background
The growth of moso bamboos has an extremely obvious phenomenon of growing in big and small years, bamboo forests in China are managed according to a big and small year system, the phenomenon of growing in big and small years of moso bamboos is considered to be related to nutritional conditions, meteorological conditions, plant diseases and insect pests of forest lands all the time, and the difference between the moso bamboos in big and small years is less concerned. On the basis, researches show that continuously eating the annual bamboo leaves in the field or the laboratory is obviously beneficial to the survival and development of important leaf-eating pests of moso bamboos, namely the bamboo moth, compared with eating the young bamboo leaves. Therefore, different pest defense measures are taken by moso bamboos in the big and small years, and the old bamboos take the measures of 'tolerance' to the leaf-eating pests, so that the pests are more likely to occur in bamboo forests in the big year.
When fed by insects, a series of changes occur in plants, such as: osmotic pressure, calcium ion channels, endogenous hormone content, etc., to achieve defense against different insects. The Jasmonic Acid (JA) signal pathway is the basis of the response process of plants to feeding insects, wherein the regulation of the transcription inhibitor JAZ protein in the jasmonic acid signal pathway is a key step. The research shows that JA is not directly involved in the regulation of the expression of plant defense genes, but is converted into JA-Ile in an activated state under the catalysis of a plurality of enzymes, and the expression of the plant defense related genes is activated through a COI1-JAZ-MYC2 complex after the JA enters a cell nucleus. A large amount of JA-Ile is firstly combined with SCF (Skip/Cullin/Fbox) protein E3 ubiquitination ligase F-box subunit COI1 protein (SCFCOI 1), and is combined with SCFCOI1 to be specifically combined with jasmonic acid ZIM functional domain protein (JAZ) so that JAZ1 is ubiquitinated or degraded under the action of 26S proteasome. The JAZ1 protein inhibits the expression of downstream plant insect-resistant genes by combining with MYC2 when not degraded, and MYC2 is combined with MED25 after degraded, so that the downstream insect-resistant genes are expressed and a large amount of secondary substances including phenols, flavonoids and tannins are synthesized. The function of the JAZ protein family is diverse and has certain differences among species. Therefore, the application of the gene in moso bamboos is beneficial to controlling the major outbreak of moso bamboo leaf pests, and has important significance for reasonable management and management of moso bamboo forests.
Disclosure of Invention
The invention aims to provide moso bamboos, aiming at the phenomenon that the moso bamboos are easy to cause insect damage in the annual management process of the moso bamboosPhJAZ1The application of the gene in the difference of insect resistance of bamboo leaves in the year of old and young. Explore the Phyllostachys pubescensPhJAZ1The gene is applied to moso bamboos in an age-level pest defense mechanism, is favorable for controlling the major outbreak of moso bamboo leaf pests, and has important significance for reasonable management and management of moso bamboo forests.
In order to achieve the purpose, the invention adopts the following technical scheme:
moso bambooPhJAZ1Gene:
Phyllostachys pubescensPhJAZ1obtaining of genes: according to Arabidopsis thalianaAtJAZ1The homologous gene sequences in the moso bamboo official website (http:// www.bamboogdb.org /) are aligned, the gene with the highest homology (PH 01000360G 1030) is selected as a template, and PCR primers are designed by virtue of Primer 5:
the forward primer SEQ ID No. 3F: 5'-ATGGAGATGTCTGCGTCCGCGA-3';
the reverse primer SEQ ID No. 4R: 5'-TTGGCTGCATTCTGTGTTCAAGC-3'.
The total RNA of the moso bamboo is extracted by taking moso bamboo leaves as raw materials. And (3) performing reverse transcription by taking the total RNA of the moso bamboo as a template to synthesize the cDNA of the moso bamboo. Using Mao bamboo cDNA as template, utilizing the above-mentioned forward primer SEQ ID No.3 and reverse primer SEQ ID No.4 to make PCR amplificationPhJAZ1A gene. And (4) recovering a PCR product, transforming after connection, and selecting a positive clone for sequencing. Obtained by sequencing and cloningPhJAZ1The gene length is 726bp, and the nucleotide sequence is shown as SEQ ID No. 1.
Moso bambooPhJAZ1The amino acid sequence of the gene coded protein is shown as SEQ ID No. 2.
Mao bamboo-containing materialPhJAZ1An overexpression vector of the gene.
Mao bamboo-containing materialPhJAZ1The construction method of the gene overexpression vector comprises the following steps: obtained by amplificationPhJAZ1Gene linked to vector carrying GFP green fluorescent protein tag by Infusion ligaseConstruction on pCambia3301PhJAZ1A gene overexpression vector.
Using moso bambooPhJAZ1Gene overexpression vector transformation of Arabidopsis thalianajaz1Mutant to obtain Phyllostachys pubescensPhJAZ1And (3) gene overexpression plants.
Moso bambooPhJAZ1The application of the gene in the difference of insect resistance of bamboo leaves in the year of old and young.
Mao bamboo-containing materialPhJAZ1The application of the gene over-expression vector in the pest resistance difference of the bamboo leaves in the year and year.
Moso bambooPhJAZ1Application of gene in pest resistance difference of young and old bamboo leaves, moso bambooPhJAZ1The gene is over-expressed, and synthesis of insect-resistant secondary metabolites in plants is inhibited.
Moso bambooPhJAZ1Application of gene in pest resistance difference of young and old bamboo leaves, moso bambooPhJAZ1The gene is over-expressed to inhibit the synthesis of tannin as an insect-resistant secondary metabolite in plants.
Moso bambooPhJAZ1Application of gene in pest resistance difference of young and old bamboo leaves, moso bambooPhJAZ1The expression level of the gene in the bamboo of the older year is higher than that of the gene in the bamboo of the younger year.
Moso bambooPhJAZ1Application of gene in pest resistance difference of young and old bamboo leaves, moso bambooPhJAZ1The leaves of the gene over-expression plants are fed with the cotton bollworm larvae, so that the survival rate of the cotton bollworm larvae can be improved, and the weight of the cotton bollworm larvae can be increased.
The invention has the advantages that: the method clones and obtains the moso bamboos related to JA signal path regulation in the response process of the moso bamboos to insect feedingPhJAZ1The gene has obvious difference in expression amount in moso bamboos of different ages, and the expression level of the gene in bamboo of old age is differentPhJAZ1The expression level is higher than that of the young bamboo. Mao bambooPhJAZ1Application of gene in pest resistance difference of young and old bamboo leaves through moso bambooPhJAZ1The gene is over-expressed, and the synthesis of tannin as an insect-resistant secondary metabolite in plants is inhibited; using moso bambooPhJAZ1The leaves of the gene over-expression plants are fed with the cotton bollworm larvae, so that the survival rate of the cotton bollworm larvae can be improved, and the weight of the cotton bollworm larvae can be increased. Mao bambooPhJAZ1The gene has an age-level pest defense mechanism, is favorable for controlling the major outbreak of moso bamboo leaf pestsThe reasonable management of the bamboo forest is of great significance.
Drawings
FIG. 1 Mao bambooPhJAZ1Gene PCR amplified electrophoretogram. Marker: DNA Marker;PhJAZ1: mao bambooPhJAZ1A gene.
FIG. 2 Mao bambooPhJAZ1The gene overexpression is carried with ACT2, and the structure of GFP-PhJAZ1 is shown schematically. ACT2 promoter: ACT2 promoter gene sequence; GFP: a green fluorescent protein gene sequence; BamH 1: a specific enzyme cutting site;PhJAZ1: mao bambooPhJAZ1A gene sequence; 35S Terminator: 35S terminator.
FIG. 3 Phyllostachys PubescensPhJAZ1Gene overexpression vector transformation of Arabidopsis thalianajaz1Western Blot result chart of mutant positive plants.PhJAZ1/jaz1#1- PhJAZ1/jaz1#5Positive plants;jaz1：arabidopsis thalianajaz1Mutation; col-0: wild type Arabidopsis thaliana Col-0.
FIG. 43 survival Rate of Heliothis armigera larvae.jaz1: feeding foodjaz1Mutant plant leaves;PhJAZ1/jaz1: feeding foodPhJAZ1/jaz1Overexpressing plant leaves; col-0: feeding leaves of wild Col-0 plants; a, B the different capitalized letters in the figure show significant differences (P＜0.01）。
FIG. 53 is a weight chart of young bollworm at age.jaz1: feeding foodjaz1Mutant plant leaves;PhJAZ1/jaz1: feeding foodPhJAZ1/jaz1Overexpressing plant leaves; col-0: feeding leaves of wild Col-0 plants; a, B the different capitalized letters in the figure show significant differences (P＜0.01）。
FIG. 6 PhJAZ1 relative expression map. A, B the different capitalized letters in the figure show significant differences (P< 0.01), different lower case letters of a, b and c represent significant differences (P＜0.05）。
FIG. 7 is a graph showing measurement of tannin content.jaz1： jaz1Tannin content in the leaves of the mutant plants;PhJAZ1/jaz1： PhJAZ1/jaz1overexpresses the tannin content in the plant leaves; col-0: tannin content in leaves of wild Col-0 plants; a, B different capital letters in the figureShows that the difference is extremely significant (P＜0.01）。
Detailed Description
EXAMPLE 1 obtaining JAZ1 loss-of-function mutants
The JAZ1 function-deficient mutant is usedjaz1Mutants, ordered from The Arabidopsis resource center (ABRC, The Ohio State University Right Hall 1060 Carmack Road Columbus, OH 43210 USA).jaz1The mutant is numbered SALK _011957, and is inserted into wild type Arabidopsis thaliana Col-0 by T-DNAjaz1The gene is deleted, thereby deleting the function of JAZ 1.
Example 2PhJAZ1Obtaining of genes
According to Arabidopsis thalianaAtJAZ1The homologous gene sequences in the moso bamboo official website (http:// www.bamboogdb.org /) are aligned, the gene with the highest homology (PH 01000360G 1030) is selected as a template, and PCR primers are designed by virtue of Primer 5:
the forward primer SEQ ID No. 5F: 5'-ATGGAGATGTCTGCGTCCGCGA-3';
the reverse primer SEQ ID No. 6R: 5'-TTGGCTGCATTCTGTGTTCAAGC-3'.
The total RNA of the moso bamboo is extracted by using a ZYMO Research Quick-RNA MicroPrep kit R1051 (Fast RNA isolation Kits for total RNA and small/miRNA from cells and soft tissue) RNA extraction kit by taking moso bamboo leaves as raw materials. Mu.g of total Mao bamboo RNA was used as a template, and the cDNA of Mao bamboo was synthesized by reverse transcription using the Prime script RT reagent Kit with gDNA Eraser (CAT: RR 047A) from Takara. PCR amplification is carried out by taking PrimeSTAR HS DNA Polymerase high-fidelity enzyme, moso bamboo cDNA as template, and forward primer SEQ ID No.5 and reverse primer SEQ ID No.6 as primersPhJAZ1A gene. The PCR reaction system is as follows: PrimeSTAR HS DNA polymerase 0.5 μ L, dNTP Mixture 4 μ L, PrimeSTAR Buffer 25 μ L, template DNA 2 μ L, forward primer SEQ ID No.5 (20 μ M) 1 μ L, reverse primer SEQ ID No.6 (20 μ M) 1 μ L, ddH2O was added to 50 to 50 . mu.L. PCR reaction procedure: 2min at 98 ℃; 10sec at 98 ℃, 15sec at 55 ℃, 1min at 72 ℃ and 30 cycles; 72 deg.C for 10min, 16 deg.CAnd + ∞. The results of the PCR are shown in FIG. 1,PhJAZ1the size of the gene target band is about 700 bp. And (4) recovering a PCR product, transforming after connection, and selecting a positive clone for sequencing. Obtained by sequencing and cloningPhJAZ1The gene length is 726bp, and the gene sequence is shown in SEQ ID No. 1; the amino acid sequence is shown in SEQ ID No. 2.
Example 3 PhJAZ1Construction of Gene overexpression vectors
Obtained by amplificationPhJAZ1The gene and the vector pCambia3301 carrying GFP green fluorescent protein label are digested with BamH1 specific nucleotidase respectively, and then Infusion ligase (In-Fusion HD Cloning Kit) is used for digesting the gene and the vectorPhJAZ1The gene was ligated to pCambia3301 vector to constructPhJAZ1A gene overexpression vector. The connecting system is as follows:PhJAZ11 μ L of gene fragment, 1 μ L of pCambia3301 vector, and 0.5 μ L of Infusion ligase; the reaction conditions are as follows: 50 ℃ for 15 min. After connection, escherichia coli DH5 alpha is transformed, after shake cultivation for 1 hour at 37 ℃, spread on an LB solid medium plate containing kanamycin resistance with the final concentration of 50 ng/mu L, after overnight cultivation, positive monoclonal bacteria are selected, PCR is identified, and then sequencing is carried out by a sequencing company, wherein sequencing primers are as follows:
SEQ ID No.7：pCambia3301(BamH1)-seqF：5’-TGCCCGACAACCACTACCTG-3’，
SEQ ID No.8：pCambia3301(BamH1)-seqR：5’-TATGCTCAACACATGAGCGAAAC-3’。
performing overnight amplification and shaking culture on positive bacteria with correct sequencing result in LB liquid culture medium containing kanamycin resistance with the final concentration of 50 ng/mu L, and extracting plasmids by using a Tiangen plasmid miniprep kit to obtainPhJAZ1The gene overexpression vector ACT2 is GFP-PhJAZ1, and the structural schematic diagram of the vector is shown in figure 2.
Example 4 transformation of plants
Plant transformation is carried out by adopting an agrobacterium-mediated floral dip method. Firstly, the constructed overexpression vector ACT2, GFP-PhJAZ1 is transferred into agrobacterium AgL0 through a freeze-thaw method to obtain agrobacterium AgL0 carrying the overexpression vector ACT2, GFP-PhJAZ 1. The agrobacterium AgL0 carrying the overexpression vector ACT2 GFP-PhJAZ1 is infected by a floral dip methodjaz1Mutants, co-infected twice, one week apart. Harvesting of transformed plant speciesScreening the seed by taking herbicide Basta as a screening marker to obtain a transformant plant PhJAZ1-jaz1. Taking wild type Arabidopsis thaliana Col-0 as a positive control, andjaz1the mutant was a negative control. Positive plants were obtained by Western Blot (protein immunoblotting) verification using PhJAZ1-GFP fusion protein as the target band, and the Western Blot results are shown in fig. 3.
EXAMPLE 5 Heliothis armigera feeding
The first-hatched 1-instar larva of bollworm is obtained from Tianjia Gonggong 807 insect-breeding chamber of Fujian agriculture and forestry university, and fed with positive transformed plant from the first day of first hatchingPhJAZ1/jaz1Leaves, death was periodically recorded daily and new leaves were fed until the first day of three instars, larvae survival and larvae weight were counted. Feeding wild type Arabidopsis thaliana Col-0 leaf as positive control groupjaz1Mutant leaf is negative control group. The feeding results are shown in fig. 4 and 5. FIG. 4 shows the results obtained withPhJAZ1/jaz1The survival rate of the cotton bollworms fed by plant leaves at the 3 rd age is obviously higher than that of the cotton bollworms fed by plant leavesjaz1The survival rate of the mutant plant leaves fed with the cotton bollworms at the 3 rd age is higher. FIG. 5 shows the results obtained inPhJAZ1/jaz1The weight of the bollworm fed by the plant leaves in 3-year age is remarkably higher than that of the bollworm fed by the plant leaves in 3-year agejaz1Mutant and Col-0 plant leaves were fed with the body weight of bollworms at 3 years of age.
Example 6 bamboo leaves of different agesPhJAZ1Determination of Gene expression level
(1) The source of bamboo leaves
The new bamboo, the old bamboo and the young bamboo leaf are all taken from the mountain village of Ou City of Fujian province, the new bamboo is the first-year-old bamboo, the second-year-old bamboo is the old bamboo, and the new bamboo is the young bamboo at intervals of one year, so that the period of the old bamboo and the young bamboo is cycled every two years.
(2) Analysis of PhJAZ1 Gene expression quantity by qPCR relative quantitative method
Total RNA from Phyllostachys pubescens leaves was extracted using ZYMO Research Quick-RNA MicroPrep Kits R1051 (Fast RNA isolation Kits for total RNA and small/miRNA from cells and soft tissue) RNA extraction kit. Each material was sampled with 1.0. mu.g of RNA, and cDNA of PhJAZ1 was obtained using Prime script RT reagent from Takarathe first strand of the cDNA was synthesized by reverse transcription using the Kit t with gDNA Eraser (CAT: RR 047A). Primer design for PhJAZ1 real-time fluorescent quantitative PCR Primer design used Primer3 (http:// Primer3plus. com/cgi-bin/dev/Primer3plus. cgi) with TIP41 (GenBank accession No. 242384689) as the internal reference gene; real-time fluorescent quantitative PCR was performed according to the instructions of the Hieff qPCR SYBR Green Master Mix (Low Rox Plus) (CAT: 11202ES 08) (Shanghai assist san Biotech Co., Ltd.) and the QuantStudio 6 Flex PCR apparatus. Four biological replicates per sample, three technical replicates were performed.PhJAZ1The gene qPCR primer is SEQ ID No.9 qPhJAZ1# 1-F: 5'-ATGGAGATGTCTGCGTCCG-3' and SEQ ID No.10 qPhJAZ1# 1-R: 5'-GGAACAGCTCCATGGTCTCC-3', respectively; the primer sequence of the reference gene TIP41 is as follows: SEQ ID No.11 TIP 41-F: 5'-AAAATCATTGTAGGCCATTGTCG-3' and SEQ ID No.12 TIP 41-R: 5'-ACTAAATTAAGCCAGCGGGAGTG-3' are provided.
The relative expression amounts of PhJAZ1 genes in the leaves of New bamboo, old bamboo and young bamboo were determined by qPCR (real-time fluorescent quantitative PCR), and the results are shown in FIG. 6. The results of FIG. 6 show that the relative expression level of PhJAZ1 gene in the annual bamboo leaves is significantly higher than that of the young bamboo leaves.
Example 7 determination of tannin
Extracting the transformed plant by referring to tannin content test box (manufacturer: Jianglai biological, product number: YX-C-A516-96)PhJAZ1/jaz1And wild type Col-0 andjaz1tannin content of the leaves of the seedlings of the mutant plants was determined by visible spectrophotometry at 760nm, and the results are shown in FIG. 7. FIG. 7 results showjaz1The tannin content in the mutant plant is obviously higher than that of the transformed plantPhJAZ1/jaz1And tannin content in wild type Col-0, indicatingPhJAZ1The over-expression of (a) inhibits the synthesis of tannin.
The above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made in accordance with the claims of the present invention should be covered by the present invention.
SEQUENCE LISTING
<110> Fujian agriculture and forestry university
Application of <120> Phyllostachys Pubescens PhJAZ1 gene in difference of insect resistance of bamboo leaves in large and small years
<130> 12
<160> 12
<170> PatentIn version 3.3
<210> 1
<211> 726
<212> DNA
<213> Gene sequence of Phyllostachys pubescens PhJAZ1 Gene
<400> 1
atggagatgt ctgcgtccgc gaggcccggg gagaggacga gcagcttcgc catggcctgc 60
agcctcctca gccgctacgt ccgccagaac ggcgccgccg cggccgagct cggcctcggg 120
atcaaaggtg aggccgagcc gcagagggcg ccggcgacga tgagcttgtc gcccggagcg 180
gagggcgagg aggcggagag gatgaaggag accatggagc tgttcccgca gagcaccggc 240
ttcggcgtgc aggatgctgc cgcccctgat gctacaaata gggagcaaga gaagcgtcag 300
ctgaccatct tctacggtgg gaaggtgctc gtgttcaacg acttcccagc caacaaggca 360
aagggcctga tgcagctggc tagcaagggc agccctgtcg tccataacgt ctctgcaccc 420
acagctgcta ctgacaacac catggtccac acggccgtgc cgtccccagc gagtagcttg 480
cctgctgtcc aggttgatgc tcagaagcct gctcgtccga atgcttctga tctgcctatt 540
gctaggaagg cgtcacttca tcgattcctt gagaagagaa aggatcgtct caatgcaaag 600
acaccatatc aagcttctcc ctcagatgca acagcagtca agaaagagcc tgagagccag 660
ccatggcttg gactagggcc gaatgccgtg aagcccaagc tgagcttgaa cacagaatgc 720
agccaa 726
<210> 2
<211> 242
<212> PRT
<213> amino acid sequence of Phyllostachys pubescens PhJAZ1 gene
<400> 2
Met Glu Met Ser Ala Ser Ala Arg Pro Gly Glu Arg Thr Ser Ser Phe
1 5 10 15
Ala Met Ala Cys Ser Leu Leu Ser Arg Tyr Val Arg Gln Asn Gly Ala
20 25 30
Ala Ala Ala Glu Leu Gly Leu Gly Ile Lys Gly Glu Ala Glu Pro Gln
35 40 45
Arg Ala Pro Ala Thr Met Ser Leu Ser Pro Gly Ala Glu Gly Glu Glu
50 55 60
Ala Glu Arg Met Lys Glu Thr Met Glu Leu Phe Pro Gln Ser Thr Gly
65 70 75 80
Phe Gly Val Gln Asp Ala Ala Ala Pro Asp Ala Thr Asn Arg Glu Gln
85 90 95
Glu Lys Arg Gln Leu Thr Ile Phe Tyr Gly Gly Lys Val Leu Val Phe
100 105 110
Asn Asp Phe Pro Ala Asn Lys Ala Lys Gly Leu Met Gln Leu Ala Ser
115 120 125
Lys Gly Ser Pro Val Val His Asn Val Ser Ala Pro Thr Ala Ala Thr
130 135 140
Asp Asn Thr Met Val His Thr Ala Val Pro Ser Pro Ala Ser Ser Leu
145 150 155 160
Pro Ala Val Gln Val Asp Ala Gln Lys Pro Ala Arg Pro Asn Ala Ser
165 170 175
Asp Leu Pro Ile Ala Arg Lys Ala Ser Leu His Arg Phe Leu Glu Lys
180 185 190
Arg Lys Asp Arg Leu Asn Ala Lys Thr Pro Tyr Gln Ala Ser Pro Ser
195 200 205
Asp Ala Thr Ala Val Lys Lys Glu Pro Glu Ser Gln Pro Trp Leu Gly
210 215 220
Leu Gly Pro Asn Ala Val Lys Pro Lys Leu Ser Leu Asn Thr Glu Cys
225 230 235 240
Ser Gln
<210> 3
<211> 22
<212> DNA
<213> Artificial sequence
<400> 3
atggagatgt ctgcgtccgc ga 22
<210> 4
<211> 23
<212> DNA
<213> Artificial sequence
<400> 4
ttggctgcat tctgtgttca agc 23
<210> 5
<211> 22
<212> DNA
<213> Artificial sequence
<400> 5
atggagatgt ctgcgtccgc ga 22
<210> 6
<211> 23
<212> DNA
<213> Artificial sequence
<400> 6
ttggctgcat tctgtgttca agc 23
<210> 7
<211> 20
<212> DNA
<213> pCambia3301(BamH1)-seqF
<400> 7
tgcccgacaa ccactacctg 20
<210> 8
<211> 23
<212> DNA
<213> pCambia3301(BamH1)-seqR
<400> 8
tatgctcaac acatgagcga aac 23
<210> 9
<211> 19
<212> DNA
<213> qPhJAZ1#1-F
<400> 9
atggagatgt ctgcgtccg 19
<210> 10
<211> 20
<212> DNA
<213> qPhJAZ1#1-R
<400> 10
ggaacagctc catggtctcc 20
<210> 11
<211> 23
<212> DNA
<213> TIP41-F
<400> 11
aaaatcattg taggccattg tcg 23
<210> 12
<211> 23
<212> DNA
<213> TIP41-R
<400> 12
actaaattaa gccagcggga gtg 23
Claims (10)
1. Moso bambooPhJAZ1The gene is characterized in that the nucleotide sequence is shown as SEQ ID No. 1.
2. Used for amplifying moso bambooPhJAZ1A primer for a gene, characterized in that: the nucleotide sequences of the primers are shown as SEQ ID No.3 and SEQ ID No.4。
3. Moso bamboo as claimed in claim 1PhJAZ1A gene-encoded protein characterized by: the amino acid sequence is shown in SEQ ID No. 2.
4. An overexpression vector comprising the gene of claim 1.
5. Moso bamboo as claimed in claim 1PhJAZ1The application of the gene in the difference of insect resistance of bamboo leaves in the year of old and young.
6. The use of the overexpression vector of claim 4 for the pest resistance difference of bamboo leaves in the year of old and young.
7. Phyllostachys pubescens as claimed in claim 5PhJAZ1The application of the gene in the insect resistance difference of the bamboo leaves in the year and the year is characterized in that: mao bambooPhJAZ1The gene is over-expressed, and synthesis of insect-resistant secondary metabolites in plants is inhibited.
8. Phyllostachys pubescens as claimed in claim 5PhJAZ1The application of the gene in the insect resistance difference of the bamboo leaves in the year and the year is characterized in that: mao bambooPhJAZ1The gene is over-expressed to inhibit the synthesis of tannin as an insect-resistant secondary metabolite in plants.
9. Phyllostachys pubescens as claimed in claim 5PhJAZ1The application of the gene in the insect resistance difference of the bamboo leaves in the year and the year is characterized in that: mao bambooPhJAZ1The expression level of the gene in the bamboo of the older year is higher than that of the gene in the bamboo of the younger year.
10. Phyllostachys pubescens as claimed in claim 5PhJAZ1The application of the gene in the insect resistance difference of the bamboo leaves in the year and the year is characterized in that: mao bambooPhJAZ1The leaves of the gene over-expression plants are fed with the cotton bollworm larvae, so that the survival rate of the cotton bollworm larvae can be improved, and the weight of the cotton bollworm larvae can be increased.
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