| Bacterial non-coding small RNA(sRNA)is a kind of nucleic acids that length is 50-500bp,which can modulate the expression of the target gene by complementary pairing with the mRNA sequence of the target gene.The sRNA of the majority of the bacteria is encoded by the 5' and 3' ends of the protein-encoding gene between the untranslated region or the gene region.sRNA is an important regulatory factor in vivo,involved in many biological activities.In order to screen the non-coding small RNAs associated with pathogenicity in Xanthomonas oryzae pv.oryzae PX099A,PX099A was cultured in the common growth medium PSA and hrp gene cluster induction medium XOM2,respectively,total RNA was extracted and sent to the company for high-throughput sequencing.For high-throughput sequencing results were analyzed,and the construct the Xoo PXO99A sRNA library of rice bacterial leaf blight.The sRNAs were divided into cis-and trans-two categories according to the coding region and the position information in the genome.Among them,cis-sRNA has 337,and trans-sRNA has 264.Compared with the expression level of sRNA in two kinds of culture,the obvious difference expression of 10 sRNA were screened out.The results of high throughput sequencing were verified by qRT-PCR,and the results were basically consistent;The target genes of differentially expressed sRNA were predicted by TargetRNA software,and their functions were preliminarily understood,for example,trans3747 might regulate cell division and membrane protein formation.10 differentially expressed sRNA molecules were obtained by comparing the expression of sRNA in the pathogenesis-related gene-inducing medium XOM2 and the common growth medium PSA by high-throughput sequencing.In this study,the sRNA gene of PXO99A was knocked out by using the method of dual-exchange of homologous arm and the gene was repaired by cDNA,and the strain of PX099A ?sRNA and C-sRNA was obtained.The pathologic function of sRNA was studied by measuring the growth rate of sRNA deletion mutants,the pathogenicity of host rice,and the allergic reaction on non-host tobacco.The results showed that the growth rate of sRNA deletion mutants was slowed down in the PSA medium compared with the wild type,and the effect of different sRNA deletion on the strain of PX099A was different.The pathogenicity of different sRNA deletion mutants on the sunny rice was also The expression of trans217 and trans3287 was significantly different from that of wild type,and trans197 and trans3287 had the same effect on the pathogenicity of rice.The results showed that trans191 had no effect on the pathogenicity of rice.The bacterial content of leaves infected with different sRNA deletion mutants was not the same as that of wild type PX099A.After sRNA was repaired,the contents of PXO99A Pathogenicity and its own growth rate;sRNA deletion mutants caused by tobacco allergic reactions are not the same,trans217 and trans3287 which can not cause allergic reactions,and trans191 and wild-type PX099A-induced allergic reactions almost no difference.The Arabidopsis cDNA libraries were screened using HpalXoo as a bait and six interacting proteins were found including plant water channel protein PIP1;4.HpalXoo,but not ANT(deletion mutant,removed HpalXoo N-terminal 53 amino acids),could interact with PIP1;4 by bimolecular fluorescence complementation and fluorescence dys observation.And the interaction obviously localized at the cell membrane.Leaf photosynthesis rates are increased and the plant growth is enhanced in contrast to the normal process without HpalXoo-AtPIP1;4 interaction.In Arabidopsis,HpalXoo-enhanced growth associates with photosynthetic physiology and is attributable to increases of mesophyll conductance(gm)to CO2 and net photosynthesis(AN)rate,indicating the functional linkage of Hpa1Xoo to PIPs that may facilitate CO2 transport. |
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