Analysis Of Four Rice Circular RNAs Gene Targeted Knockout Mutants

Circular RNA?circRNA?is a type of covalently closed long-chain non-coding RNA that has no free ends.It is connected to the"end"of the 5'upstream sequence through the3'downstream sequence to form a circular structure,which is not easy due to its special structure.It is cleaved by exonuclease to maintain high stability.Circular RNA plays an important regulatory role in biological processes,and research on circular RNA in plants is still scarce.In this study,four circular RNAs?Os02circ25329,Os06circ02797,Os03circ00204,Os05circ02465?were used to knock out the mutants.The agronomic characters of the mutants,abiotic stress,parent/flanking gene expression and transcriptome analysis were used to preliminary analyze the four Biological functions of rice circular RNAs genes.The main research contents and results are as follows:1.The identification of homozygous mutant materials without vectors showed that Os02circ25329 obtained two genotype materials-578bp/-578bp?pZJP053-03-03,pZJP053-27-01?and-584bp/-584bp?pZJP053-29-16?,Os06circ02797 obtained two genotype materials-328bp/-328bp?pZJP054-07-01-21?and-332bp/-332bp?pZJP054-32-03?,Os03circ00204 obtained-684bp/-684bp?pZJP055-76-04?and-661bp/-661bp?pZJP055-78-01?A variety of genotype materials,Os05circ02465 obtained-843bp/-843bp?pZJP057-23-06?and-799,?1bp/-799,?1bp?pZJP055-78-04?two genotype materials.2.The results of agronomic trait analysis showed that:Os02circ25329,Os06circ02797,Os03circ00204,Os05circ02465 four circular RNAs mutants showed no significant difference in particle length,particle width and thousand kernel weight from WT.3.The results of abiotic stress treatment showed that the seeds of Os02circ25329,Os06circ02797 and Os03circ00204 were sensitive to salt during seed germination,and the seeds of Os05circ02465 mutant were salt-tolerant;The four circular RNAs mutants showed the same performance as WT under salt,Cu²⁺and drought?PEG?stress.4.The results of parent/flanking gene expression analysis showed that:Os02circ25329 and Os06circ02797 knockouts did not affect the normal expression of parental genes,Os03circ00204 targeted knockout significantly increased the left gene expression,Os05circ02465 knockout did not affect normal expression of flanking genes.5.Transcriptome analysis showed that Os02circ25329 affected the differential expression of 70 genes and 4 miRNAs,Os06circ02797 affected the differential expression of 135 genes and 15 miRNAs,Os03circ00204 affected the differential expression of 98 genes and 7 miRNAs,Os05circ02465 affected 54 Differential expression of genes and 2 miRNAs.