CN108849890A - Application of the rice sesquiterpene synthases TPS46 gene in terms of preventing and treating pink rice borer - Google Patents

Abstract

The invention discloses application of the rice sesquiterpene synthases TPS46 gene in terms of preventing and treating pink rice borer.The invention also includes a kind of pink rice borer attractant, the pink rice borer attractant includes the transgenic paddy rice after the rice sesquiterpene synthases TPS46 gene or its overexpression.Pink rice borer attractant of the present invention also includes the mixture of (E)-β-farnesene, cresotinic acid, (E)-α-bergmot oil alkene, (E)-β-carypohyllene, α-Humuleno, linalool, α-bisabolene and limonene.First identified of the present invention goes out a kind of overexpression in can significantly lure the abiogenous rice TPS46 gene of pink rice borer on rice plant, field experiments have shown that harm of the pink rice borer for non-transgenic control rice can be significantly reduced in the rice for overexpressing the TPS46 gene, be beneficial to pink rice borer field trap and integrated control.

Description

Application of the rice sesquiterpene synthases TPS46 gene in terms of preventing and treating pink rice borer

Technical field

The invention belongs to field of biotechnology, and in particular to rice sesquiterpene synthases TPS46 gene is in terms of preventing and treating pink rice borer Application, be more particularly to TPS46 gene transgenic overexpression crop after and its regulation mixture new application.

Background technique

During long-term coevolution, for the feeding harm for resisting pest, plant has gradually formed a set of complicated effective Defense system.After pest is caused harm, plant can enhance the specific herbivore-induced volatile (Herbivore-Induced of release Plant Volatiles, HIPVs), on the one hand these HIPVs can directly resist pest, including poisoning or repellent action, separately On the one hand it can be identified and be utilized by natural enemy, important work is waved in the indirect defenses of plant as " emergency " signal of plant again With.In recent years, effect and its molecular regulation mechanism of the HIPVs in host plant-pest-natural enemy tritrophic interactions, and The excavation of its application potential is pest-important research direction of natural enemy chemical ecological regulation.Rice is that the first generalized grain of China makees Object, eating rice production is to ensuring that China's grain security is of great significance.For a long time, striped rice borer (Chilo suppressalis Walker) and the harm of the snout moth's larvas such as pink rice borer (Sesamia inferens Walker) is always main insect pest in Rice Production Problem seriously affects rice good quality and high output and stable yields, and seriously limits the volume increase potential of rice.Currently, chemical prevention is snout moth's larva The main preventions in worm field, but the generation of drug resistance is densely largely accelerated using insecticide, and cause society The strong worry for the problems such as public is for grain security and environmental pollution.In addition, just brill moth in the short time after snout moth's larva egg hatching Rice stem, and its entire larval phase and pupa time are completed in rice straw, cause conventional biological or chemical insecticide anti-to it It controls extremely difficult.Therefore, there is an urgent need to develop go out efficient, low toxicity, prevent and treat bio-safety and environmental-friendly snout moth's larva in production Measure.

Plant sesquiterpene synthases TPS family is a median size gene family and terpenes volatile matter biosynthesis Key gene is usually made of 20-150 gene.In recent years, with the development and plant of sequencing and Protocols in Molecular Biology The successive announcement of object genomic data, a variety of plants such as arabidopsis, rice, grape, tomato, selaginella tamariscina, apple, poplar and eucalyptus TPS family gene is furtherd investigate in object.

Summary of the invention

Goal of the invention：Based on above-mentioned problem, we specify rice TPS46 expression quantity by two by experimental study Change snout moth's larva damage induction and significantly rises (p later<0.05), and transgenosis is constructed using transgenosis overexpression technology to overexpress TPS46 rice, apparent tiller later period-jointing stage overexpression TPS46 rice and its specific volatile matter mixture are compared to control Pink rice borer oviposition and naturally-occurring brill moth (p can significantly be attracted<0.05).And our results of study show (E)-β- farnesene、methyl salicylate、(E)-α-bergamotene、(E)-β-caryophyllene、α-humulene、 Eight kinds of volatile matter mixtures such as linalool, α-bisabolene and limonene are the high tables of TPS46 overexpression rice specificity The volatile matter combination reached.Therefore, the development of transgenic technology and molecular chemistry ecology technology makes us for turning using TPS46 Integrated control provides a kind of completely new strategy in turn for trans-genetic hybrid rice and its product regulation snout moth's larva behavior, and therefore, the present invention is wanted The technical issues of solution the new opplication there is provided rice sesquiterpene synthases TPS46 gene in terms of pink rice borer prevention and treatment.

Technical solution：It is big in prevention and treatment that in order to solve the above-mentioned technical problems, the present invention provides rice sesquiterpene synthases genes Application in terms of snout moth's larva.

The sesquiterpene synthases gene order for deriving from rice is gene TPS46, the cDNA sequence overall length of the gene 1641bp encodes 546 amino acid.

Wherein, the application includes the spy that the rice sesquiterpene synthases TPS46 gene is regulated and controled in overexpression rice Application of the anisotropic metabolite in terms of preventing and treating pink rice borer.The gene regulates and controls in the plant that overexpression rice plant lures pink rice borer 8 kinds of metabolites.The cDNA sequence of gene TPS46 gene such as SEQ ID NO：Shown in 1, the amino acid sequence such as SEQ ID NO：Shown in 2.

Wherein, the specific metabolic product is (E)-β-farnesene ((E)-β-farnesene), methyl Salicylate (cresotinic acid), (E)-α-bergamotene ((E)-α-bergmot oil alkene), (E)-β-caryophyllene ((E)-β-carypohyllene), α-humulene (α-Humuleno), linalool (linalool), α-bisabolene (α-bisabolene) And it is one or more of in limonene (limonene).

Preferably, the specific metabolic product is (E)-β-farnesene ((E)-β-farnesene), methyl Salicylate (cresotinic acid), (E)-α-bergamotene ((E)-α-bergmot oil alkene), (E)-β-caryophyllene ((E)-β-carypohyllene), α-humulene (α-Humuleno), linalool (linalool), α-bisabolene (α-bisabolene) And the mixture in limonene (limonene).

The positive rice (Oe strain) of TPS46 transgenosis overexpression and its 8 species specific volatile matter products of the invention Cultivate, identification be detailed in article that we deliver early period (Sun et al., 2017, Frontiers in plant science, 3: 8-110), more than 15 Oe strains are obtained.

The specific metabolic product regulated and controled in overexpression rice about the rice sesquiterpene synthases gene is being prevented and treated The concrete measure of application in terms of pink rice borer is as follows：

(1) on the basis of building early period TPS46 transgenosis overexpression rice, selection growth T2 is for transgenic positive water Rice；

(2) T2 is planted for TPS46 transgenic positive rice in field at random, determines the table of different lines rice TPS46 gene The amount of influence up to to(for) pink rice borer natural incidence (%)；

(3) plantation T2 connects worm and tests clear different lines rice for 3-4 age pink rice borer for TPS46 transgenic positive rice The selectivity of larva influences；

(4) for field planting T2 for TPS46 transgenic positive rice, cage determines different lines rice for the pink rice borer of release The influence of egg laying amount.

The content of present invention further includes a kind of pink rice borer attractant, and the pink rice borer attractant includes the rice sesquiterpene synthases Product after TPS46 gene or its overexpression.

Wherein, the pink rice borer attractant includes that the rice sesquiterpene synthases TPS46 gene is adjusted in overexpression rice The specific metabolic product of control.

Wherein, the specific metabolic product is (E)-β-farnesene ((E)-β-farnesene), methyl Salicylate (cresotinic acid), (E)-α-bergamotene ((E)-α-bergmot oil alkene), (E)-β-caryophyllene ((E)-β-carypohyllene), α-humulene (α-Humuleno), linalool (linalool), α-bisabolene (α-bisabolene) And it is one or more of in limonene (limonene).

Preferably, the pink rice borer attractant includes (E)-β-farnesene ((E)-β-farnesene), methyl Salicylate (cresotinic acid), (E)-α-bergamotene ((E)-α-bergmot oil alkene), (E)-β-caryophyllene ((E)-β-carypohyllene), α-humulene (α-Humuleno), linalool (linalool), α-bisabolene (α-bisabolene) And the mixture in limonene (limonene).

Beneficial effect：First identified of the present invention goes out a kind of overexpression in can significantly lure pink rice borer to send out naturally on rice plant Raw rice TPS46 gene, field experiments have shown that pink rice borer can be significantly reduced for non-in the rice for overexpressing the TPS46 gene The harm of non-transgenic control rice, be beneficial to pink rice borer field trap and integrated control.

Detailed description of the invention

Fig. 1 T2 is for different lines rice plant of tillering stage TPS46 gene expression amount test map, Oe6, Oe7, Oe9, Oe 11 TPS46 transgenosis overexpresses rice；OWt non-transgenic control strain rice；

The natural incidence (%) of Fig. 2 different lines Rice ragged stunt virus, Oe6, Oe7, Oe9, Oe11 are the super table of TPS46 transgenosis Up to rice；OWt non-transgenic control strain rice；

Fig. 3 different lines rice attracts percentage for 3-4 age pink rice borer larva, and Oe6, Oe7, Oe9, Oe 11 turns for TPS46 Gene overexpresses rice；OWt non-transgenic control strain rice；

Cage investigates pink rice borer Adult worms producting eggs amount on Fig. 4 different lines rice, and Oe6, Oe7, Oe9, Oe11 are TPS46 transgenosis Overexpress rice；OWt non-transgenic control strain rice.

Specific embodiment

Below with reference to specific embodiment, the present invention is further explained.It should be understood that these embodiments are merely to illustrate this hair It is bright rather than limit the scope of the invention.It should also be understood that after reading the content taught by the present invention, art technology Personnel can make various changes or modifications the present invention, and such equivalent forms equally fall within application the appended claims and limited Range.

Unless stated otherwise, the present invention uses reagent, method and apparatus for the art conventional reagent, method and are set It is standby.

Embodiment 1

1, the cultivation and plantation of T2 generation positive TPS46 transgenosis overexpression rice：

The cultivation that T0 overexpresses positive rice (Oe strain) for TPS46 transgenosis is detailed in the article (Sun that we deliver early period et al.,2017,Frontiers in plant science,3:8-110), more than 15 Oe strains are obtained, wherein Oe6, 7, the overexpression effect of 9 and 11 strains is best, is the main experimental materials of the application, while Oe false positive strain is as OWt pairs According to.T0, T1 and T2 use sterile water vernalization in illumination box for transgenosis overexpression TPS46 rice paddy seed first, are catalyzed Condition is 22-28 DEG C of temperature, (daytime photoperiod：Night) 16h：8h, relative humidity 50 ± 5%.Then, Oe6, Oe7, Oe9, Oe11 And each plant single-strain planting of OWt strain is placed among the greenhouse of Jiangsu Province Agriculture Science Institute (24-36 DEG C) in plastic tub, Each strain plants 50 basins (13 centimetres of 11 cm x height of diameter under upper 14 cm x of diameter).T0, T1 are planted for transgenic positive The strain plantation complete generation simultaneously collects seed, and T2 carries out expression quantity detection and related experiment for transgenic paddy rice length to tiller later period.

Conclusion：T2 is smoothly obtained for tillering stage Oe6, Oe7, Oe9 and Oe11 to be measured of transgenosis overexpression TPS46 gene Rice strain.

2, T0, T1 and T2 are detected for the expression quantity of transgenosis overexpression TPS46 trans-genetic hybrid rice：

T2 uses SV for the Total RNAs extraction of tiller later period Oe6, Oe7, Oe9, Oe11 and OWt system rice leaf and leaf sheath Total total serum IgE separation system (Promega company, operating procedure is referring to instructions book), concentration application NanoDrop 1000 ultramicrospectrophotometers are detected (NanoDrop company).Then the synthesis of cDNA template originates in 1 μ g total serum IgE, Carry out related experiment (Promega company) using MMLV reverse transcriptase, carrying out RNase H processing after synthesis rapidly, (TaKaRa is public Department), and application 1000 ultramicrospectrophotometer of NanoDrop detection starting cDNA concentration.Quantitative fluorescent PCR tests (qPCR) Agents useful for same uses SYBR Premix Ex Taq Kit (TaKaRa company), and instrument is iCycler iQ (Bio-Rad company), Analysis software is version 3.0a (Bio-Rad company).Quantitative fluorescent PCR is reacted with RNase-free water (TaKaRa company) For negative control, originating cDNA concentration is about 80-100ng, 5 repetitions of each sample, and reaction condition is 3 footworks, recurring number 40 It is a.TPS46 gene expression amount qPCR detection primer sequence be TPS46-F (5 '-TGAAGAGGCACTAGGTCCAAAC-3 ') and TPS46-R(5′-CCATCCCAACTAAAGAAGCACA-3′).With rice housekeeping gene EF-1 α (Genbank Serial No.: It AK061464) is internal standard gene, sequence is EF-1 α-F (5 '-AGACGCACATCAACATCG-3 ') and EF-1 α-R (5 '- GAACTTCCACAGGGCAATA-3′).Primer uses 5.0 software design of Primer, and is had by the raw work bioengineering share in Shanghai The synthesis of limit company combining unit.The TPS46 gene that transgenosis overexpression rice strain Oe6, Oe7, Oe9, Oe11 are compareed relative to OWt Relative expression quantity changes with 2^-ΔΔCtMethod calculates (Livak and Schmittgen, 2001), the significance of difference of relative expression quantity Analysis is using the DuncanShi duncan's new multiple range method in statistical software SAS (SAS v 9.0).

Conclusion：As shown in Figure 1, tillering stage Oe6, Oe7, Oe9 and Oe11 rice strain to be measured that T2 is overexpressed for transgenosis The expression quantity of middle TPS46 gene is significantly higher than OWt control strain (p<0.05), the expression quantity of transgenic line TPS46 gene is extremely Reach 200 times or more of control strain less.

3, T2 is for 8 species specificity volatile matter quantitative test of tiller later period Oe and OWt strain rice TPS46

T2 for tiller later period Oe positive rice (Oe6,7,9,11) and OWt strain rice single plant be respectively placed in two it is identical Cylindrical glass cylinder (diameter × height：25cm × 50cm) among, every 1 basin of cylinder, remaining port sealing in addition to inlet, outlet port.Greatly Gas sampling instrument regulates and controls air-flow 1ml/min (atmosphere sampling instrument QC-3 type buy in Beijing City Inst. of Labour Safety Science), with containing After having an adsorption column absorptive collection 6h of Tenax-TA adsorption stuffing 60mg (filler is bought in Shanghai ANPEL Scientific Instruments Corporation), N-hexane 300 μ l of the adsorption column containing internal standard ethyl caprate 2.586ng is rinsed.When volatile materials acquisition time is Beijing Between 11:00-17:00, test is repeated 3 times.8 kinds of T2 are used for TPS46 overexpression rice specificity volatile matter quantitative test to be compared Two-phase gas chromatography mass spectrometry system GCxGC-ToF (Pegasus4D type, LECO company of the U.S.), sample volume 1ul, temperature program and 8 kinds of spies The retention of anisotropic volatile matter be detailed in article early period (Sun et al., 2017, Frontiers in plant science, 3: 8-110), Volatility Levels are repeated 3 times by inner mark method ration, test, and multiple using 9.0 statistical software Duncan formula of SAS Compare the analysis for carrying out volatile substance content difference.T2 is waved for tiller later period Oe positive rice and 8 species specificity of OWt strain rice See Table 1 for details for the content of hair object.

Conclusion：As shown in table 1,8 species specificity volatile matters in the tillering stage Oe rice strain that T2 is overexpressed for transgenosis (E)-β-farnesene, cresotinic acid, (E)-α-bergmot oil alkene, (E)-β-carypohyllene, α-Humuleno, linalool, α-opopanax Alkene and limonene are all remarkably higher than OWt control strain (p<0.05), show that this 8 kinds of volatile matters are T2 for transgenosis overexpression Oe water Rice mainly functions substance.

4, the natural parasitic rate (%) of field pink rice borer：

T2 is randomly placed for tiller later period Oe6, Oe7, Oe9, Oe11 and OWt strain rice positive single plant in agriculture section of Jiangsu Province Institute experimental plot, individual line per treatment select 5 plants of positive rice, are repeated on the different location in experimental plot, each strain for five times 25 plants, all rice are 125 plants total, and to investigate field insect pest, a situation arises.24 DEG C -32 DEG C of field temperature, the photoperiod is close 14h：10h, for relative humidity close to 50%, what periphery pest worm sources were verified has pink rice borer S.inferens and brown paddy plant hopper N.Lugens Deng.The test data sheet of pest parasitic rate is in daily 15:00-17:00, and instituted an inquiry from the tillering stage of rice and extend to jointing Phase.

Conclusion：As shown in Fig. 2, Field Pests naturally-occurring percentage the result shows that, T2 for transgenosis overexpress TPS46 Pink rice borer natural incidence is significantly higher than the parasitic rate of OWt control on the jointing stage of gene Oe6, Oe7, Oe9 and Oe11 rice to be measured (p<0.05), the incidence of transgenosis overexpression strain pink rice borer is at least up to 80%, and OWt control strain only has 20% or less. The rice that this result discloses transgenosis overexpression TPS46 gene can significantly attract pink rice borer parasitism (p naturally compared to control< 0.05)。

5, selection percentage (%) of the interior 3-4 age pink rice borer larva S.inferens for different rice strains：

T2 is crosslinked with OWt rice leaf respectively for tiller later period Oe6, Oe7, Oe9 and Oe11 strain single plant, fixed.Then Single head 3-4 age pink rice borer S.inferens larva, is placed between two plants of rice.Experiment (every plant of Oe strain rice, 1 plant of OWt every time Rice), 20 3-4 age pink rice borer S.inferens larvas are tested, are repeated three times, 60 test worms are amounted to.5 rice of test are planted every time Strain, Oe the and OWt strain more renewed, indoor air temperature is maintained at 26 DEG C -28 DEG C when test, the raw test of pink rice borer S.inferens larva It tests and is carried out in daily 10:00-16:00.

Conclusion：As shown in figure 3, significantly having a preference for TPS46 transgenosis overexpression rice compared to OWt control 3-4 age pink rice borer larva Strain (p<0.05).The rice that this result discloses transgenosis overexpression TPS46 gene can significantly attract pink rice borer compared to control 3-4 instar larvae (p<0.05).

6, egg laying amount of the cage measurement pink rice borer S.inferens on different rice strains：

T2 is for tiller later period Oe6, Oe7, Oe9 and Oe11 positive rice each 1 plant and the total eight plants of tillers of four plants of OWt system rice Later period rice random alignment is placed in cuboid cage (80 mesh gauzes, 80 cm x of cage length and width are 120 centimetres high).For examination Pink rice borer larva is collected in academy of agricultural sciences of Jiangsu Province rice test field, is raised indoors with the insect box equipped with wild rice stem to (reference of pupating Korea Spro is superfine, and 2012, applied entomology report, 49：The method of 281-285), the adult after 10 pairs of emergence is transferred in mating cage, to Its of short duration mating is released in cage to its oviposition, and raises and supply its feeding with 10% hydromel, and test is repeated 5 times.Oviposition Afterwards, by produce have ovum blade and leaf sheath mark, to Adult worms producting eggs after, by production there is the blade of ovum and leaf sheath to cut, then will Tissue with ovum, which is placed in the glass culture dish (diameter 9cm) for being covered with wet filter paper, carries out moisturizing, investigates egg laying amount under microscope.

Conclusion：As shown in figure 4, pink rice borer cage egg laying amount statistical result shows T2 for transgenosis overexpression TPS46 gene Pink rice borer egg laying amount is significantly higher than OWt control (p on tiller later period Oe6, Oe7, Oe9 and Oe11 rice to be measured<0.05).This result is taken off Show that the rice of transgenosis overexpression TPS46 gene can significantly attract pink rice borer Adult worms producting eggs (p compared to control<0.05).

In production, we pass through pink rice borer Incidence investigation the result shows that：Due to abiogenous dispersibility, lead to pink rice borer field Relative difficulty is prevented and treated using chemical pesticide and natural enemy, therefore produces and is badly in need of identifying the product that can significantly trap pink rice borer. TPS46 gene mainly its metabolite that works functions, therefore its 8 kinds of metabolites play a significant role.The present invention The result shows that TPS46 transgenic paddy rice and its 8 kinds of metabolites of specificity can significantly lure the naturally-occurring of pink rice borer, can Significantly to protect maternal control rice, and it can largely trap the concentration harm of pink rice borer.This gain effect shows that the present invention has Important potential using value, and significantly the trapping for pink rice borer initial population in production, and then concentrate biology, chemical synthesis Prevention and treatment is of great significance.

Sequence table

<110>Jiangsu Province Agriculture Science Institute

<120>Application of the rice sesquiterpene synthases TPS46 gene in terms of preventing and treating pink rice borer

<160> 6

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1641

<212> DNA

<213>Rice sesquiterpene synthases gene (rice terpene synthases)

<400> 1

atgtcatcga cacctgcagc taatttctcc aatgaagacg atgagcgaaa ggctcctacc 60

ggcttccacc ccagcctctg gggcgatttc ttcatcagtt accagccacc tactgcacct 120

aagcatgctt acatgaaaga aagggctgaa gtgctaaaag aagaagttag aaaggtagta 180

aagggctcaa atgaagtacc agagatattg gatcttgtga tcacgctgca acgacttgga 240

ttggatagtt actacaaggc cgagatagat gagcttctct gcactgttta caacaccgac 300

tacaacgata aagatttaca cctagtttct cttcgatttt accttctacg gaagaacggc 360

tatgatgtgt catctgacat atttcaacat tttaaagaca aagaggggag ttttgttgct 420

gatgacacaa gaagtctttt aagcttatat aatgcggcat atatgaggac tcatggtgaa 480

aaagtacttg atgaagcagt tgttttcact actaaccgcc taagatcaga attgaaacat 540

ttgaaatctc cggtagccga tgaagtgtct cttgctcttg atacaccact attccggagg 600

gttagaataa tagaaacaca aaactatatt cctatttatg aaagcgcgac tacacgaaat 660

gaagccatat tagaatttgc aaagttgaat gtcaatctcc ttcaacttat ttattgtgag 720

gagttaaaaa ctatcacacg atggtggaaa gagcttaacg ttgaatccaa cttaagcttc 780

attcgggata gaatagtgga aatgcatttt tggatgacag gagcatgctc ggagccccat 840

tattctcttt tacgaattat acttacaaag atgacagcat ttatcaccat tctagatgat 900

atatttgata catatgctac aaccgaggag agcatgatgc ttgccaaagc gatatatatg 960

tgcaatgaat ctgcaacagt tctacttcca aaatacatga aggatttcta cttatattat 1020

ttaaagacat ttgattcatt tgaagaggca ctaggtccaa acaaaagtta tcgagtgctt 1080

tatttcaaag agctgttcaa gatattaatt aaaggatact ctgaagagat aaaatggcgt 1140

gatgatcatt acattccaaa aacaatagag gaacacctgg aactttccag aatgactgtt 1200

ggtgcctttc aactagcatg tgcttcttta gttgggatgg gtgattttat aacagaggat 1260

actctcgatt atcttttgac ttatccaaaa cttattaagt cttacacaac atgtgtacgg 1320

ctctcaaatg atattgcttc aacaaagcgt gaacaagcag gggaccacta tgcttccaca 1380

attcagtgtt acatgctgca gcatggtaca acaatacatg aagcatgcat tgggataaag 1440

gagttgatag aagattcgtg gaaggatatg atgaaagaat accttgcacc aacaaacctt 1500

cagccaaaaa tcgtggcaag aacagttatt gattttgcac gaactgggga ttatatatac 1560

aagcaagcag attcattcac cttttcacat acaatcaagg atatgatagc atcactttac 1620

gttgagccat atagcattta a 1641

<210> 2

<211> 546

<212> PRT

<213>Rice sesquiterpene synthases gene (rice terpene synthases)

<400> 2

Met Ser Ser Thr Pro Ala Ala Asn Phe Ser Asn Glu Asp Asp Glu Arg

1 5 10 15

Lys Ala Pro Thr Gly Phe His Pro Ser Leu Trp Gly Asp Phe Phe Ile

20 25 30

Ser Tyr Gln Pro Pro Thr Ala Pro Lys His Ala Tyr Met Lys Glu Arg

35 40 45

Ala Glu Val Leu Lys Glu Glu Val Arg Lys Val Val Lys Gly Ser Asn

50 55 60

Glu Val Pro Glu Ile Leu Asp Leu Val Ile Thr Leu Gln Arg Leu Gly

65 70 75 80

Leu Asp Ser Tyr Tyr Lys Ala Glu Ile Asp Glu Leu Leu Cys Thr Val

85 90 95

Tyr Asn Thr Asp Tyr Asn Asp Lys Asp Leu His Leu Val Ser Leu Arg

100 105 110

Phe Tyr Leu Leu Arg Lys Asn Gly Tyr Asp Val Ser Ser Asp Ile Phe

115 120 125

Gln His Phe Lys Asp Lys Glu Gly Ser Phe Val Ala Asp Asp Thr Arg

130 135 140

Ser Leu Leu Ser Leu Tyr Asn Ala Ala Tyr Met Arg Thr His Gly Glu

145 150 155 160

Lys Val Leu Asp Glu Ala Val Val Phe Thr Thr Asn Arg Leu Arg Ser

165 170 175

Glu Leu Lys His Leu Lys Ser Pro Val Ala Asp Glu Val Ser Leu Ala

180 185 190

Leu Asp Thr Pro Leu Phe Arg Arg Val Arg Ile Ile Glu Thr Gln Asn

195 200 205

Tyr Ile Pro Ile Tyr Glu Ser Ala Thr Thr Arg Asn Glu Ala Ile Leu

210 215 220

Glu Phe Ala Lys Leu Asn Val Asn Leu Leu Gln Leu Ile Tyr Cys Glu

225 230 235 240

Glu Leu Lys Thr Ile Thr Arg Trp Trp Lys Glu Leu Asn Val Glu Ser

245 250 255

Asn Leu Ser Phe Ile Arg Asp Arg Ile Val Glu Met His Phe Trp Met

260 265 270

Thr Gly Ala Cys Ser Glu Pro His Tyr Ser Leu Leu Arg Ile Ile Leu

275 280 285

Thr Lys Met Thr Ala Phe Ile Thr Ile Leu Asp Asp Ile Phe Asp Thr

290 295 300

Tyr Ala Thr Thr Glu Glu Ser Met Met Leu Ala Lys Ala Ile Tyr Met

305 310 315 320

Cys Asn Glu Ser Ala Thr Val Leu Leu Pro Lys Tyr Met Lys Asp Phe

325 330 335

Tyr Leu Tyr Tyr Leu Lys Thr Phe Asp Ser Phe Glu Glu Ala Leu Gly

340 345 350

Pro Asn Lys Ser Tyr Arg Val Leu Tyr Phe Lys Glu Leu Phe Lys Ile

355 360 365

Leu Ile Lys Gly Tyr Ser Glu Glu Ile Lys Trp Arg Asp Asp His Tyr

370 375 380

Ile Pro Lys Thr Ile Glu Glu His Leu Glu Leu Ser Arg Met Thr Val

385 390 395 400

Gly Ala Phe Gln Leu Ala Cys Ala Ser Leu Val Gly Met Gly Asp Phe

405 410 415

Ile Thr Glu Asp Thr Leu Asp Tyr Leu Leu Thr Tyr Pro Lys Leu Ile

420 425 430

Lys Ser Tyr Thr Thr Cys Val Arg Leu Ser Asn Asp Ile Ala Ser Thr

435 440 445

Lys Arg Glu Gln Ala Gly Asp His Tyr Ala Ser Thr Ile Gln Cys Tyr

450 455 460

Met Leu Gln His Gly Thr Thr Ile His Glu Ala Cys Ile Gly Ile Lys

465 470 475 480

Glu Leu Ile Glu Asp Ser Trp Lys Asp Met Met Lys Glu Tyr Leu Ala

485 490 495

Pro Thr Asn Leu Gln Pro Lys Ile Val Ala Arg Thr Val Ile Asp Phe

500 505 510

Ala Arg Thr Gly Asp Tyr Ile Tyr Lys Gln Ala Asp Ser Phe Thr Phe

515 520 525

Ser His Thr Ile Lys Asp Met Ile Ala Ser Leu Tyr Val Glu Pro Tyr

530 535 540

Ser Ile

545

<210> 3

<211> 22

<212> DNA

<213>Artificial sequence TPS46-F (Artificial Sequence)

<400> 3

tgaagaggca ctaggtccaa ac 22

<210> 4

<211> 22

<212> DNA

<213>Artificial sequence TPS46-R (Artificial Sequence)

<400> 4

ccatcccaac taaagaagca ca 22

<210> 5

<211> 18

<212> DNA

<213>Artificial sequence EF-1 α-F (Artificial Sequence)

<400> 5

agacgcacat caacatcg 18

<210> 6

<211> 19

<212> DNA

<213>Artificial sequence EF-1 α-R (Artificial Sequence)

<400> 6

gaacttccac agggcaata 19

Claims (7)

1. application of the rice sesquiterpene synthases TPS46 gene in terms of preventing and treating pink rice borer.

2. application according to claim 1, which is characterized in that the application includes rice sesquiterpene synthases TPS46 gene Application of the specific metabolic product regulated and controled in overexpression rice in terms of preventing and treating pink rice borer.

3. application according to claim 2, which is characterized in that the specific metabolic product is (E)-β-farnesene, first One in base salicylic acid, (E)-α-bergmot oil alkene, (E)-β-carypohyllene, α-Humuleno, linalool, α-bisabolene and limonene Kind is several.

4. application according to claim 2, which is characterized in that the application specifically includes following steps：

(1) on the basis of the TPS46 transgenosis of building early period overexpresses rice, selection growth T2 is for TPS46 transgenic positive Rice；

(2) T2 is planted for TPS46 transgenic positive rice in field at random, determines the expression of different lines rice TPS46 gene Amount, and apparent TPS46 overexpresses influence of the positive rice for pink rice borer natural incidence；

(3) plantation T2 connects worm and tests clear different lines rice for 3-4 age pink rice borer larva for TPS46 transgenic positive rice Selectivity influence；

(4) for field planting T2 for TPS46 transgenic positive rice, cage determines that different lines rice produces the pink rice borer of release The influence of ovum amount.

5. a kind of pink rice borer attractant, which is characterized in that the pink rice borer attractant is synthesized comprising rice terpenes described in claim 1 Product after enzyme TPS46 gene or its overexpression.

6. pink rice borer attractant according to claim 5, which is characterized in that the pink rice borer attractant includes the rice terpenes The specific metabolic product that synzyme TPS46 gene is regulated and controled in overexpression rice.

7. pink rice borer attractant according to claim 6, which is characterized in that the pink rice borer attractant includes (E)-β-Fa Ni Alkene, cresotinic acid, (E)-α-bergmot oil alkene, (E)-β-carypohyllene, α-Humuleno, linalool, α-bisabolene and limonene One or more of.