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Exploration And Functional Analysis Of Odorant Degrading Enzymes Related Genes In Green Plant Bug,Apolygus Lucorum

Posted on:2013-07-22Degree:MasterType:Thesis
Country:ChinaCandidate:C ShenFull Text:PDF
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Heteroptera:Miridae belongs to the Hemiptera blindPentatomidae, is an important pest in cotton production in China,causing huge losses every year all over the world cotton production.Various cotton-growing areas in China have been found in cottonblind bugs more than 20 species, of which the more serious types ofharm, lucorum, black bugs and alfalfa plant bugs, three-point bugsand tarnished plant bug. With the extensive planting of Bt cotton in1997, effectively control the hazards of the the bollworm mainLepidoptera pests, cotton Miridae populations occur to theincreasing number of secondary pest of cotton rose to one of themajor pests. Lucorum has become the most serious of dominantpopulations in China’s Yellow River, the Yangtze River Valleycotton-growing areas.To combat the bugs mainly rely on chemicalpesticides, which not only easily lead to the Miridae drug resistance,but also huge pollution and damage caused to the environment,which makes the development of green and environment-friendlyprevention lucorum new methods and new technology seemsparticularly important. At present, the sense of smell to interferewith the target insects to its effective control is the control ofinsect behavior hot spots. Insect identification spouse release the sex pheromone or plantvolatile odor related protein located in the insect olfactory sensilla,including odor-binding proteins (odorant binding proteins, OBPs)and chemosensory proteins (chemosensory proteins, CSPs)olfactory receptors (olfactory receptors, Ors), odor-degradingenzyme odorant degrading enzymes (ODEs) and the olfactory nerveyuan membrane protein (sensory neuron membrane. proteinsSNMPs). Insect olfactory responses, including a series ofconsecutive reactions: a combination of scent molecules, odorantmolecules in the sensilla lymph transport odor molecules andreceptor combined with the conversion of the signal transductionand termination. , plays a very important role in signal terminationin the olfactory response, not only to avoid olfactory organs bycontinuous chemical stimulation also reduces the interference ofthe signal saturation.Lucorum, Lygus lucorum antennae cDNA library by SMARTtechnology was successfully constructed and detailedbioinformatics analysis to identify a large number ofolfactory-related genes, filter out the smell of degradation-relatedgenes, the result was glutathione -S-transferase (GST), genesequences for a total of 27 esterase (EST) gene sequences of 12, 31cytochrome P450 reductase gene sequences. SMARTer RACEtechniques combined with cloning-related odor degrading enzymes(ODEs) gene was successfully cloned the full-length sequence of anew the lucorum esterase gene, a total length of 1810bp openreading frame of 1713bp encoding 571 amino acids. Of thissequence named for Aluc EST1, Gene bank accession number JQ715614. Predict the molecular weight of molecular weight63.44Kd, isoelectric point (PI) as 5.99. Laid the foundation forbiological function of the lucorum odor degrading enzymes.Subsequently cloned two lucorum reference gene, and RT-PCRsemi-quantitative and real-time QPCR technique analysis Aluc EST1expression profiles in various tissues. The study found that thetentacles of the esterase gene in lucorum, head, chest, abdomen,feet, wings various parts have expression,which is the highestexpression in the chest and abdomen, the gene abundantlyexpressed in the antennae. Interaction between enzyme andsubstrate for determination of enzyme activity, the study ofalpha-naphthyl acetate as substrate, fast blue RR salt aschromogenic reagent reaction, determination of enzyme activity inAluc EST1 is 1.224868 (m OD min-1 mg-1). Through the substratetheα-/β- naphthyl acetate, lucorum esterase map analysis and foundthat in the lucorum adult, you can see two clear bands, and thesame bar band in the lucorum nymphs can be also seen where thebands appear darker in the male, showing the enzyme activity inmales may be higher.The above studies laid a theoretical foundation for the system toclarify the lucorum sense of smell to identify the molecularmechanism of the prevention and the treatment of the lucorum .This may become an important theoretical basis for furtherprevention and control field lucorum.
Keywords/Search Tags:Lygus lucorum Meyer-Dür, Odor degradingenzymes, Aluc EST1esterase, Expression profiling, enzyme activity, Esterase isozyme electrophoresis
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