Molecular Cloning Of The Rhyzopertha Dominica And Antennal Sensilla Of Sitophilus Granarius,Tribolium Castaneum And Their Host Range

Rhyzopertha dominica,Tribolium castaneum and Sitophilus granarius are most important stored food insect pests in the world,and have serious threat to grain production.Predominantly chemical pesticide control measures,has caused such problems as pest resistance increase and pollution.Olfactory play important role during the host plants of insect,and if we could clear the olfactory receptor mechanism of these pests in the host location,it will provide a theoretical basis for using the olfactory behavior regulation of these insect pests.In the current research,the effect of broken grain seed on reproduction of Sitophilus granarius,Rhyzopertha dominica and Tribolium castaneum were studied,the characteristic of antennae sensilla of Sitophilus granarius(L.)and Tribolium castaneum were studied by transmission electron microscopy(TEM).And also,the olfactory relevant protein genes,CSPs and PBPs of Rhyzopertha dominica,were cloned,and the expression levels of PBPs1 and PBPs2 were checked by q PCR.The main results are as follows:1.Effect of grain cracked on reproduction of Sitophilus granarious,Rhyzopertha dominica and Tribolium castaneum on different hostgrainsThe effects of different host grains and the broken grains on the development,life cycle,sex ratio,adult weight and population of Sitophilus granarius,Rhyzopertha dominica and Tribolium castaneum were studied.The results showed that the total number of S.granarius F1 adult beetles emergence on wheat was much higher than those from maize,barley and rice,while the number of R.dominica and T.castaneum were similar on four different grains.The number of three pest females emergence on four grains were higher than the numbers of males.The adults weight of three pests emerged on diverse hostgrains were similar.2.Ultrastructural observations on antennal sensilla of Sitophilus granariusThe ultra structure of antennae sensilla of Sitophilus granarius(L.)were studied by transmission electron microscopy(TEM).The results of TEM micrographs showed that sensilla basiconica of S.granaries are characterized by strongly corrugated pores around the cuticle,while micrographs of sensilla coeloconica longitudinal sections showed smooth-surfaced pegs and flat-tipped manner an apical pore indicated that they role in gustatory function.TEM micrographs of sensilla chaetica longitudinal sections showed dendrite branches and cuticular pore arrow heads that possibly occupied in the observation of moisture,heat and carbon dioxide.These studies are providing information play important roles of the sensilla types in the host location behavior of S.granarius.3.Ultrastructural observations on antennal sensilla of Tribolium castaneumThe ultra structure of antennae sensilla of Sitophilus granarius(L.)Tribolium castaneum(Coleoptera: Tenebrionidae)were studied by transmission electron microscopy(TEM).Transmission electron microscopy of the antennal sensilla of Tribolium castaneum showed sensilla trichodea to have a thick cuticular wall and regular point-shaped pores around the cuticular wall.Sensilla basiconica possessed a thin wall and continuous pores and dendrites.The pores on the walls of the sensilla indicate that they participate in a function in sensing chemical stimuli and in olfactory functions.4.Molecular cloning of CSP2 s,PBP1 and PBP2 genes of Rhyzopertha dominicaBase on the information of antenna c DNA library of R.dominica,three olfactory protein genes,including Rhydomica CSPs2,Rhydomica PBPs1,Rhydomica PBPs2 genes(GenBank accessions are KJ186798.1,KJ186830.1,KJ186831.1 separately.),were successfully cloned.For sequencing and phylogenetic analysis Rhydomica CSPs1 and Rhydomica CSPs2 belonged to Minus-C CSPs showed that have 4 conserved cysteine residues,while Rhydomica PBPs1 and Rhydomica PBPs2 showed conserved amino acids in all PBPs six conserved cysteine residues.The results of transcription expression level of PBPs1 and PBPs2 of R.dominica showed that the expression level of Rhydomnica PBP2 is much higher than that of Rhydomnica PBP1.The variation transcription level at the different developmental time suggested the PBP1,and PBP2 had their particularly job in searching food sources,mates and oviposition sites.