RESUMO
Hymenopteran parasitoids generally show a haplo-diploid sex determination system. Haploid males are produced from unfertilized eggs, whereas diploid females develop from fertilized eggs (arrhenotokous). In some cases, diploid females develop from unfertilized eggs (thelytokous). Diglyphus wani (Hymenoptera: Eulophidae) is a biological control agent for agromyzid leafminers and have arrhenotokous and thelytokous strains. However, the morphological characteristics of two strains of D. wani are so similar that it is difficult to accurately distinguish them based on morphology. Here, a rapid molecular identification method was developed based on the mitochondrial gene cytochrome c oxidase I (COI) and one-step multiplex PCR. Two primer combinations, PC1 (Ar-F1/Th-F1/WR2) and PC2 (Ar-F1/Th-F4/WR2), were designed and repeatedly screened to distinguish two strains simultaneously, of which two special forward primers Th-F1/Th-F4 were used for the thelytokous strain and one special forward primer Ar-F1 was used for the arrhenotokous strain. In addition, a common reverse primer, WR2, was used for both strains. The PC1 and PC2 PCR assays were effective in distinguishing the two strains at different developmental stages and field colonies. This method provides a reliable, highly sensitive, and cost-effective tool for the rapid identification of the two strains of D. wani.
Assuntos
Himenópteros , Vespas , Feminino , Masculino , Animais , Himenópteros/genética , Reação em Cadeia da Polimerase Multiplex , Agentes de Controle BiológicoRESUMO
Diglyphus species are ecologically and economically important on agromyzid leafminers. In 2018, a thelytokous species, Diglyphuswani Liu, Zhu & Yefremova, was firstly reported and described. Subsequently, the arrhenotokous D.wani were discovered in Yunnan and Guizhou Provinces of China. We compared the morphological characteristics of thelytokous and arrhenotokous strains. However, the females of two strains had a strongly similar morphology and showed subtle differences in fore- and hind-wings. The difference was that forewing of arrhenotokous female was with denser setae overall, showing that costal cell with 2 ~ 4 rows of setae on dorsal surface and the setae of basal cell with 15 ~ 21 hairs and forewing of thelytokous female was with two rows of setae on dorsal surface and basal cell with 10 ~ 15 hairs generally. The setation beneath the marginal vein of the hind-wing of arrhenotokous female is denser than the same area of thelytokous female. To explore the genetic divergence between thelytokous and arrhenotokous strains of D.wani, the mitochondrial and nuclear gene were applied and sequenced. The polygenic analyses revealed that two strains can be distinguished by COI, ITS1 and ITS2. The mean sequence divergence between the two strains was 0.052, 0.010 and 0.007, respectively. Nevertheless, the 28S gene was unfeasible due to its containing a sharing haplotype between different strains. The two strains of D.wani are dominant parasitoids against agromyzid leafminers and such effective discernible foundation provides future in-depth studies on biological characteristics, along with insight into field application of two strains of D.wani.
RESUMO
Diglyphus species are dominant biocontrol agents for suppressing outbreaks of agromyzid leafminers in fields. In July 2015, we collected a new thelytokous species of Diglyphus in Qinghai, China. The wasp is here named as D. wani Liu, Zhu & Yefremova sp. nov., based on morphological and molecular analyses. The life history and biocontrol potential of the wasp were studied in the laboratory and by providing Liriomyza sativae larvae. The intrinsic rate of increase, finite rate of increase, and mean generation time were 0.2373 d-1, 1.2678 d-1, and 15.9 d, respectively. The wasps showed three types of host-killing behaviour, namely parasitism, host feeding and host stinging, resulting in 47.6, 94.4 and 3.4 host larvae killed per wasp over a lifetime, respectively. The finite total host-killing rate was 1.0071 and the wasp showed strong synovigeny. The wasp may be a crucial biocontrol agent against agromyzid leafminers.
