Molecular Identification Of Interspecific Differences And Male Polymorphism Of Fig Wasps Based On DNA Sequence

In this paper, we reproducted the sequence of mitochondrion cytochrome oxidase I (COI), cytochrome b (Cytb) and internal transcribed spacer2(ITS2) in two species of Ficus altissima wasps(Eupristina altissima and Eupristina sp.), Ficus virens two species of unpollinating wasps(Camarothorax bimasculinus and Otitesella minima) and Ficus superba unpollinating wasps(Otitesella ako). Using the methods of base composition analysis, base substation statistics, genetic distance calculations and building phylogenetic tree to analysis the results of sequences. The results were as follows:1. The analysis results of COI sequences were as follows:①Base composition analysis:Eupristina genus wasps both female and male showed that the average content of GC were24.2%and24.7%;The average content of GC in COI gene sequences of Ficus virens wasps (C. bimasculinus and O. minima) and Ficus superba unpollinating wasps (O. ako) were22.0%23.4%、22.4%respectively. The average content of GC in different samples showed lower than AT;②Base substation analysis showed that the main base substation of the COI sequences in two species Eupristina genus wasps were transversion, although in O. minima wasps and O. ako wasps the main base substation were transition, in C. bimasculinus wasps transversion and transition are equality.③The average K2P distance between the female wasps of E. altissima and E. sp.was0.0969, and the average K2P distance between the male wasps of the two species was0.1086, was as10times as the two groups separately; the average K2P distance among the four species of O. minima male wasps was0.0253; the average K2P distance among the four species of O. ako male wasps was0.0145; and the average K2P distance between the two species of C. bimasculinus male wasps was0.0154. The average K2P distance among the different intraspecific male morphs are slight, and there were no more than6.8%.(4)Using NJ and UPGMA methods to analyze:The female and male E. altissima clustered into the same lineage first and then clustered with E. sp. Ultimately clustered with Ficus microcarpa pollinating wasps (E. verticillata). The female and male O. minima and O. ako clustered into the same lineage respectively and then clustered together because all of them belong to Otitesella genus. They clustered with group Walkerella micrcoarpae from Ficus microcarpa finally. The male and female of C. bimasculinus from Ficus virens clustered into the same lineage showed that the genetic relationship between them was close.2. Cytb sequences analysis were as followed:①Base composition analysis: Eupristina genus wasps both female and male showed that the average content of GC were25.8%and26%;the average content of GC in O. minima, O. ako and C. bimasculinus were22.1%、22.3%and24.4%respectively. The average content of GC in different samples showed lower than AT;(2)On the bases substation analysis:Eupristina genus wasps of two females and two males Cytb bases substation is given priority to transversion; Three species O. minima, O. ako and C. bimasculinus of male and female wasps the substation of Cytb sequences are given priority to transition.③The average K2P distance between the female wasps of E. altissima and E. sp. was0.1514, and the average K2P distance between the male wasps of the two species was0.1701, both of them were6%higher; The average K2P distance among the four species of O. minima male wasps was0.0166; The average K2P distance among the four species of O. ako male wasps was0.004; and the average K2P distance between the two species of C. bimasculinus male wasps was0. All of them were6%lower.④Using NJ and UPGMA methods to analyze:The female and male E. altissima and E. sp. clustered into the same lineage respectively first, and then the wasps of E. sp. clustered with the wasps of E.verticillata from Ficus microcarpa. Ultimately the wasps of Eupristina genus clustered together; The clustering results of three kinds of the female and male wasps(O. minima、 O. ako、C. bimasculinus)were consistent with the analysis COI conclusion above.3. Ribosome ITS2analysis results:①Base composition analysis:In the female and male of Eupristina genus wasps the ribosome ITS2fragments the average content of GC were53.7%and53.6%, both of them were slightly50%higher; the average content of GC in O. minima, O. ako and C. bimasculinus were56.1%、56.7%and51.1%respectively, all of them were higher than AT.②Base substation analysis showed that two females and two malesof Euprislina genus wasps in ITS2sequences the transition rate and the rate of transversion were equal. In O. minima wasps the main base substation were transversion although in C. bimasculinus and O. ako wasps transversion and transition were equality.③The average K2P distance between the female wasps of the two species of Eupristina genus was0.1380, and the average K2P distance between the male wasps of the two spices was0.1344; The average K2P distance among the four species of O. minima male wasps was0.0284; The average K2P distance among the four species of O. ako male wasps was0.0103; and the average K2P distance between the two species of C. bimasculinus male wasps was0.0132.④Using NJ and UPGMA methods to analyze:The clustering results were consistent with the analysis COI conclusion above.The results of the COI、Cytb、ITS2showed that the two species wasps of Eupristina genus which lived in Ficus altissima were already reach the degree of diversity, and the diversity is interspecific variation; the different morphology O. minima, O. ako and C. bimasculinus male wasps were male polymorphism.