The gypsy moth—Lymantria dispar (Linnaeus)—is a worldwide forest defoliator and is of two types: the European gypsy moth and the Asian gypsy moth. Because of multiple invasions of the Asian gypsy moth, the North American Plant Protection Organization officially approved Regional Standards for Phytosanitary Measures No. 33. Accordingly, special quarantine measures have been implemented for 30 special focused ports in the epidemic areas of the Asian gypsy moth, including China, which has imposed great inconvenience on export trade. The Asian gypsy moth and its related species (i.e., Lymantria monocha and Lymantria xylina) intercepted at ports are usually at different life stages, making their identification difficult. Furthermore, Port quarantine requires speedy clearance. As such, it is difficult to identify the Asian gypsy moth and its related species only by their morphological characteristics in a speedy measure. Therefore, this study aimed to use molecular biology technology to rapidly identify the Asian gypsy moth and its related species based on the consistency of mitochondrial DNA in different life stages. We designed 10 pairs of specific primers from different fragments of the Asian gypsy moth and its related species, and their detection sensitivity met the need for rapid identification. In addition, we determined the optimal polymerase chain reaction amplification temperature of the 10 pairs of specific primers, including three pairs of specific primers for the Asian gypsy moth (L. dispar asiatic), four pairs of specific primers for the nun moth (L. monocha), and three pairs of specific primers for the casuarina moth (L. xylina). In conclusion, using our designed primers, direct rapid identification of the Asian gypsy moth and its related species is possible, and this advancement can help improve export trade in China.
Associations between Sternorrhyncha insects and intracellular bacteria are common in nature. Mealybugs are destructive pests that seriously threaten the production of agriculture and forestry. Mealybugs have evolved intimate endosymbiotic relationships with bacteria, which provide them with essential amino acids, vitamins, and other nutrients. In this study, the divergence of five mealybugs was analyzed based up the sequences of the mitochondrial cytochrome oxidase I (mtCOI). Meanwhile, the distinct regions of the 16S rRNA gene of primary symbionts in the mealybugs were sequenced. Finally, high‐throughput sequencing (HTS) techniques were used to study the microbial abundance and diversity in mealybugs. Molecular phylogenetic analyses revealed that these five mealybugs were subdivided into two different clusters. One cluster of mealybugs ( Dysmicoccus neobrevipes , Pseudococcus comstocki , and Planococcus minor ) harbored the primary endosymbiont “ Candidatus Tremblaya princeps,” and another cluster ( Phenacoccus solenopsis and Phenacoccus solani ) harbored “ Ca . Tremblaya phenacola.” The mtCOI sequence divergence between the two clusters was similar to the 16S rRNA sequence divergence between T. princeps and T. phenacola . Thus, we concluded that the symbiont phylogeny was largely concordant with the host phylogeny. The HTS showed that the microbial abundance and diversity within P. solani and P. solenopsis were highly similar, and there was lower overall species richness compared to the other mealybugs. Among the five mealybugs, we also found significant differences in Shannon diversity and observed species. These results provide a theoretical basis for further research on the coevolution of mealybugs and their symbiotic microorganisms. These findings are also useful for research on the effect of symbiont diversity on the pest status of mealybugs in agricultural systems.
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