Research On The Role Of Arabidopsis Thaliana CircR11208 In Resisting Biotic And Abiotic Stresses

Circular RNAs(circRNAs)are a class of endogenous noncoding RNAs without 5' end caps and 3' poly(A)tails,which are widely present in eukaryotic cells and are conservative and stable.circRNAs perform multiple biological functions in biological processes.There have been some investigations on circRNAs in plants,but not many,and little is known about their functions in resisting biotic and abiotic stresses.So it is very important to predict,identify and study the function of circRNAs in resisting biotic and abiotic stresses.In this study,we identified circR11208 and formation mechanism obtained by using high-throughput sequencing and bioinformatics analysis on the Arabidopsis thaliana by PCR and Sanger sequencing,we build the relevant transgenic plants,which laid a good foundation for the research of circR11208.Phenotype observation and study function,founding that circR11208 had little effect on the growth of Arabidopsis thaliana plants,but participated in the resistance process to Botrytis cinerea in Arabidopsis thaliana,and negatively regulate the growth of lateral roots of Arabidopsis thaliana under salt stress.Specific research results are as follows.1.The successful construction of circR11208 transgenic plants.In this study,the predicted circR11208 was firstly verified by PCR using RNase R treatment and poly(A)enriched RNA as the template by designing an divergent primers,and the correctness of the prediction was preliminarily determined,and further Sanger sequencing confirmed that it was an exon backsplicing product formed by non-classical splicing signals recognition.Secondly,vector of circR11208 overexpression and silencing was constructed,and transgenic plants were obtained by Agrobacterium tumefaciens mediated dip-flower transformation and used Colombian wild-type as the background.Moreover,the expression level of circR11208 was detected in the transgenic plants,and it was confirmed that the constructed vector did affect the expression of circR11208,and the homozygous transgenic plants were obtained.Phenotypic observation showed that circR11208 did not affect the growth of Arabidopsis thaliana.The acquisition of transgenic plants laid a good foundation for the functional research of circR11208 in the later stage.2.circR11208 is involved in the resistance process to Botrytis cinerea in Arabidopsis thaliana and negatively regulates the growth of lateral roots of Arabidopsis thaliana under salt stress.In this study,according to the parent gene involved in salt stress,drought stress,programmed cell death and basal resistance to Botrytis cinerea in Arabidopsis thaliana,it is speculated that circR11208 may also have certain biological functions.Therefore,the plants were infected with Botrytis cinerea,it was found that overexpression of circR11208 transgenic plants can increase the resistance of Arabidopsis thaliana to Botrytis cinerea under low spore infection conditions.The HR response and ion permeability related to programmed cell death were detected,and the correlation between circR11208 and programmed cell death was not significant.The measurement of callose and reactive oxygen species accumulation showed that circR11208 could not regulate the immune response of Arabidopsis thaliana to pathogenic bacteria.Under salt stress and drought stress treatment,and it was found that circR11208 negatively regulated the lateral root growth of Arabidopsis thaliana under salt stress,but had little correlation with drought stress.The results indicate that circR11208 plays an important role in resisting biotic and abiotic stresses,and is closely related to plant stress resistance.It enriches the functional research content of circRNAs and provides clues for the in-depth study of circRNAs.