Functional Analysis Of 33E05,34B08 And Hg-exp-1 Gene From The Soybean Cyst Nematode(Heterodera Glycines) And Research On Molecular Detection Technology Of Sugarbeet Cyst Nematode(Heterodera Schachtii)

Soybean cyst nematode(Heterodera glycines)is a devastating disease of soybean.This disease has a large range of hosts and distribution,diversity of disease transmition,resulting in serious economic losses.It’s a recognized soybean disease that is extremely difficult to control.Chemical control is the main way to prevent soybean cyst nematode.Through the study of the feeding tube related gene,identify the formation mechanism and functionary mechanism of the feeding tube,will provide an important theoretical basis for the development of new prevention and control technologies.In this study,the 33E05 gene of soybean cyst nematode was researched.The expression characteristics of 33E05 gene was analyzed by developmental expression analysis and subcellular localization.The in vitro RNAi research was carried out to explore the function of the 33E05 gene.Subcellular localization assays revealed that 33E05 and 34B08 was localized in the cell membrane,the localization experiment of protoplasts determined that it was located in the cell membrane but not the cell wall,it was speculated that the effector proteins 33E05,34B08 secreted by soybean cyst nematode may participate in the formation and maintenance of the nematode feeding tube by acting on the cell membrane and endoplasmic reticulum,etc.Gene expression analysis showed that 33E05 genes transcriptions were expressed effectively at all the developmental stage of soybean cyst nematode,and the highest expression was found in pre-parasitic and parasitic second-stage juveniles.Speculate 33E05 genes may play an important role in the process of the feeding tube formation of soybean cyst nematode.The function of 33E05 and 34B08 was studied by in vitro RNAi.qRT-PCR showed that the transcription level of target genes of second-stage juveniles was significantly decreased after dsRNA soaking treatment.In the soybean inoculation experiment,the number of nematodes in host root has no significant difference between soaking treatment and the control.It was speculated that the gene family may be related to the development of nematodes and had no effect on the nematode infection ability.Based on the preceding studies,Functional analysis of soybean cyst nematode expansion protein gene hg-exp-1 was carried out.After Hg-exp-1 gene was silenced,the number of nematodes infected in soybean root and females decreased by 38.3% and 43.4% than control.The result proved that Hg-exp-1 played an important role in the early parasitism of soybean cyst nematode.Sugar beet cyst nematode(Heteradera schachtii)is an important disease source in the production of beet,and it is also an important pathogen of entry quarantine in China.In this experiment,the samples of sugar beet cyst nematode were extracted by DNA and the RAPD primers were used for PCR amplification and sequencing,finally we designed SCAR primers specific to sugar beet nematode,which can identify sugar beet nematode in the molecular level rapidly and sensitively,and the specific fragment could be clearly identified when the dilution was 1/40000 of a cyst or 1/320 of a J2 for all replicates.It is beneficial to the rapid and effective plant quarantine in China and is also beneficial to the prevention and control of the nematode disease in China.