Molecular Mechanisms Of Chemical Senses In Apolygus Lucorum (Meyer-Diir)

With the commercial planting of BT cotton in China, Apolygus lucorum and other sucking insectsdominate as the major pests of cotton. It was meaningful of agricultural and economic to understand theinformation transmission between A.lucorum and cotton. Olfactory and gustatory are the main way ofA.lucorum to get the information of cotton.Use molecular biology to resolve the olfactory and gustatorymechanisms in molecular level has become a hot spot. In this study, by means of molecular biology thechemical senses mechanisms researchs were carried out in A.lucorum, the main results are as follows:1. Proboscises and legs cDNA library were created from3day old male and female.2. Following bioinformatics analysis of EST sequences of the library, identified four odorant bindingproteins (AlucOBP3-6)(GenBank accession number: JN573221-JN573224) and four chemosensoryproteins (AlucCSP1-4)(GenBank accession number: JN573217-JN573220).3. we quantitatively analyzed the expressions of these eight genes in different tissues of male and female byreal-time PCR and found that, AlucOBP3and AlucCSP1specifically expressed in female antennae,AlucOBP4AlucCSP2and AlucCSP3were specifically in female wing,AlucOBP5only expressed in thefemale abdomen and male leg(male leg> female abdomen), AlucOBP6had high expressions in maleproboscis and wings (proboscises> wings), AlucCSP4were high expression levels in male wings and alsoexpressed in other tissues.4. AlucOBP3-6were expressed using prokaryotic expression system, and then purified using GSTrap FF.The Binding characteristics of AlucOBPs to58kinds of chemical molecules and5kinds of cottonsecondary metabolites were further studied using bis-ANS as fluorescent probes. The study showed thatAlucOBP3could bind to benzyl alcohol, benzyl acetate, phenethyl acetate, ethylbenzene, limonene oxide,limonene oxide, linalool, tannin (Ka=16.1±1.2μM)and rutin (Ka=26.2±1.8μM); AlucOBP4bound withphenethyl alcohol,3-hexenol, ethyl butyrate, butyric acid-trans-2-hexene ester, hexyl butyrate, heptylacetate, methyl salicylate, tannin (Ka=12.3±0.3μM), catechin (Ka=12.3±0.1μM), quercetin (Ka=19.7±0.3μM), and rutin hydrate (Ka=16.43±0.9μM); AlucOBP5bound with phenylacetaldehyde (Ka=33.1±2.1μM),1-hexanol(Ka=21.4±1.2μM),cis-3-hexenal(Ka=32.7±2.4μM),cis-2-hexen-1-ol,naphthalene,heptylacetate,trans-2-hexenylbutyrate,betaionone(Ka=34.5±2.2μM),gossypol(Ka=3.2±0.1μM),tannins(Ka=3.4±0.2 μM), quercetin(Ka=7.2±0.8μM), rutin hydrate(Ka=10.3±1.5μM); AlucOBP6bound with benzyl acetate,phenylethyl acetate, carvone, linalool, eucalyptol, indole, gossypol (Ka=7.5±0.5μM), tannin (Ka=10.1±0.4μM), catechin (Ka=51.5±2.3μM), quercetin (Ka=17.9±0.9μM), and rutin hydrate (Ka=15.2±1.2μM).5. AlucCSP1-4were expressed using prokaryotic expression system, and then purified using GSTrap FF.The Binding characteristics of AlucCSPs to58kinds of chemical molecules and5kinds of cottonsecondary metabolites were further studied using bis-ANS as fluorescent probes. The study showed thatAlucCSP1could bind to geranyl acetate, butyrate-trans-2-hexene ester, catechin,gossypol (Ka=13.4±1.4μM), tannin (Ka=32.7±2.4μM), quercetin (Ka=19.7±0.7μM), and rutin hydrate (Ka=38.5±1.5μM);AlucCSP2bound with trans-2-hexenal,2-heptanone, alpha-ionol, hexyl butyrate, heptyl acetate,cis-3-hexenylhexanoate,gossypol(Ka=39.4±2.4μM), quercetin(Ka=31.8±0.1μM), tannins(Ka=33.6±2.1μM),rutin hydrate(Ka=37.9±1.8μM);AlucCSP3bound with phenylacetaldehyde,benzylalcohol,(R)-(+)-Limo-nene,gossypol(Ka=26.3±2.4μM),quercetin(Ka=35.5±2.2μM),tannins(Ka=33.6±2.1μM),rutinhydrate(Ka=44.9±1.8μM);AlucCSP4bound with gossypol(Ka=16.8±1.5μM), quercetin(Ka=35.5±0.9μM), tannins, rutinhydrate, catechin.