Effects Of Hemocyanin’s Genetic On Embryonic Development In The Migratory Locust

The migratory locust Locusta migratoria (Linnaeus) is widespread pest that isdistributed in Europe, Asia, Africa and Australia. It is widely distributed and frequentlyoutbreaked in China, also caused serious influence for agricultural production andpeople’s life. The key period for locust outbreaks is in embryonic period, however, inthe embryonic period of locusts, tracheal system has not yet formed. Hemocyanin asone of three kinds of respiratory function protein, bear the oxygen transfer function.RNA interference (RNAi) as a kind of specific inhibition mechanism of geneexpression in the body, its technology has become one of the main methods of genefunction research insects. In order to explore the locust hemocyanin function, our studyestablished the migratory locust embryo microinjection technology system, using atechnique called RNA interference to explore the locust hemocyanin gene function.Hemocyanins are respiratory proteins exiting in the hemolymph of arthropods andmolluscs which contain copper atoms. Most researches of arthropod hemocyaninfocused on the hemocyanin from Crustacean, Myriapoda, Merostomata andArachnoidea. Considering hexapods possess a tracheal system that enables thetransport of oxygen to the inner organs, respiratory proteins have long been ignored.However hemocyanins were found to be broadly existent in insecta recently.We changed typical non-holometabolous insects-locusta migratoria-as experimental subjects. In this study, we used the RNA interference method combinedwith hypoxia and an improved method of microinjection (survival80%after injected),to surpress the expression of locust hemocyanin subunit1. By real-time PCR detected,the difference is outstanding in two days, four days, even sis days after injectHC1dsRNA. Morphologicalobservation showed that compared with the control group,the treatment group embryonic development slowed down, the proportion of abnormalincreased and hatchability came down.Embryonic development showed that the locustegg’s development more slowed down under hypoxia. By comparision with the controlgroup, he treatment group slowed down than development period, some eggs oftreatment group stayed in the early blastokinesis period. This indicate that thehemocyanin may participate the embryonic development of locust.Based on our preliminary findings that hemocyanin with oxygen-carryingcapacity in the terrestrial insect locusta migratoria. By using the RNA interferencemethod, we provide the proof the the hemocyanin participated in the process of locustembryo’s development, and lay the foundation to further research about respiratoryprotein and trachea formation, embryonic development and insect hypoxia adaptation.