Identification And Distribution Of Sensory Neuron Member Protein In Dalia Antiqua

Delia antiqua(Meigen)is also can be called "onion fly".It is one of improtant Chinese underground pests.The larvae is called "garlic maggot" that is similar with the larvae of B.odoriphaga but larger parasites.Larvae drills and eats the underground part of the host plant,causing root rot,serious condition the garlic maggot can eat all of the underground part,leading plant death.The onion fly belong the underground pests,feeding the roots.It is different to prevent and control.In the long term,using the chemical pesticide bring more and more disadvantages not only increase the cost of planting,but also easily lead to environmental pollution.Insects perceive the sensitize pheromones of the same species incets and host plant volatiles released by the sensitive sense of smell.Using of insect olfactory function to control pests has been practiced on a variety of insects.1.Cloning the Sequence of DantSNMP2 analyzing the transcriptome of Onion fly,we got four fragments of DantSNMP2.According to these fragments,designing their premers primers,augmenting to got a sequence about 1000 bp.The results showed that the length of the fragment was 1192 bp,which contained the 5 ’ terminus sequence of the open reading frame of the onion fly,but the 3’ terminus was still unknown.To obtain the complete open reading frame of the onion fly,the the 3’ terminus fragment of the DantSNMP2 was amplified by RACE,getting the 775 bp fragment.Splicing two pieces of the genes,and finally getting full onion fly SNMP2(DantSNMP2)open reading frame.The DantSNMP2 open reading frame is consists of 1551 bases,and can be translated into 516 amino acid residues,with a molecular weight of 58.22 kD and a theoretical isoelectric point of 8.51.Searching the sequences of SNMP1 and SNMP2 from Culex quinquefasciatus and Bactrocera dorsalis and comparing with the DantSNMP2,the results showed that the five SNMPs from three different species contained 6 conserved cysteines participate,which can form three disulfide bonds.Using the obtained DantSNMP2 with the SNMP from other 36 kings of insects to construct phylogenetic trees.Dant SNMP2 is clustered together with other SNMP2.Analyzing the hydrophilia,the results showed that DantSNMP2 contained two hydrophobic regions and ahydrophilic cyclic structure,which proving that DantSNMP2 is a double transmembrane protein.2.Analyzing the sequence of DantSNMP2 By analyzing the hydrophily of DantSNMP2,the result showed that DantSNMP2 contained two hydrophobic regions and a hydrophilic cyclic structure,which confirmed that DantSNMP2 was a double transmembrane protein.Selecting the sequences of SNMP1 and SNMP2 from Culex quinquefasciatus and Bactrocera dorsalis were compared with DantSNMP2,the results showed that these five SNMPs contain6 conserved cysteine residues.We speculated that the 6 cysteine residues may be involved in formatting three disulfide bond,capable of stabilizing the spatial structure of Dant SNMP2.DantSNMP2 and 36 SNMPs to construct phylogenetic trees.The results showed that the SNMPs of insects were classified into two groups: SNMP1 and SNMP2.The DantSNMP2 was clustered together with the SNMP2 of other Diptera insects,relatively distant relationship with Lepidoptera.3.The distribution of DantSNMP2.2.To study the expression of DantSNMP2 in female and male tissues and larvae by RT-PCR.The results showed that DantSNMP2 is expressed in the larvae and adult.The expression patterns of DantSNMP2 is showed the same condition in different tissues of the female and male adults.The expression of DantSNMP2 was not detected in the head,chest,abdomen and wings.Diptera insects olfactory sense is main distributed in the antenna,meanwhile also distributing in the foot.The distribution of DantSNMP2 is consistent with olfactory sensilla.