Differential Analysis Of MicroRNA Of Bursaphelenchus Xylophilus Cultured At Two Temperatures And Study On The Function Of Mog-2 Gene

Bursaphelenchus xylophilus is the pathogen causing pine wood nematode disease and is listed as a worldwide quarantine nematode.High fecundity is an important invasion mechanism of B.xylophilus,and temperature is an important factor affecting various phenotypes of B.xylophilus.It is worth studying how the regulatory factor microRNA of B.xylophilus regulates genes in response to temperature and thus affects various phenotypes of B.xylophilus.In this experiment,the microRNA characteristics of B.xylophilus cultured at15℃ and 25℃were analyzed,and the function of the sex-determining gene mog-2 was analyzed.The main results are as follows:There were significant differences in the characteristics of microRNAs of B.xylophilus cultured at 15℃ and 25℃.Among the 198 microRNAs identified in B.xylophilus,18 microRNAs were only expressed in nematodes cultured at 15℃ ambient temperature,and 37 A variety of microRNAs were only expressed in nematodes cultured at an ambient temperature of 25℃,and there were 3 microRNAs that were significantly differentially expressed in B.xylophilus cultured at 15℃ and 25℃.Target gene predictions for these three microRNAs were carried out,and corresponding candidate target genes GO enrichment analysis showed that these three microRNAs with significant differences were mainly involved in mitochondrial biological processes,such as mitochondrial transmembrane transport,mitochondrial transport,and protein targeting mitochondrial processes.It can be seen that temperature can affect the expression of microRNA in B.xylophilus,and the microRNA shows obvious temperature response characteristics.Predict the microRNAs that regulate the sex-determining genes of B.xylophilus,and analyze and predict the functions of the genes regulated by them.The functions of these sexdetermining genes are mainly concentrated in cell component differentiation,germ stem cell differentiation,cell activity,sex determination,metabolic activity,cell component Formation,postembryonic morphogenesis,signal transduction,spermatogenesis,etc.Analysis of the significantly down-regulated microRNA and the gene pathway regulated by lgi-mi R-34 at 25℃ showed that lgi-mi R-34 could regulate the B.xylophilus spliceosome pathway.In this pathway,the sex-determining gene mog-2 is in the m RNA precursor.plays an important role in the splicing process.Through mog-2 RNAi,it was found that the interference of mog-2 can cause slow peristalsis of B.xylophilus,affecting the feeding of B.xylophilus,resulting in a longer growth cycle,a decrease in male-to-female ratio,and distortion of the uterus and uterine sac.The RNAi effect is heritable and appears to have some effect on the subphenotype,but the F4 phenotype effect disappears.It is considered that mog-2 is involved in the peristaltic behavior,feeding behavior,growth and development of B.xylophilus,and has an effect on the male-to-female ratio of B.xylophilus,and plays an important role in the development of B.xylophilus uterus and scrotum.