Analysis Of Phenotype And Genome And Studies On Detection Method Of Wohlfahrtiimonas Chitiniclastica

Wohlfahrtiimonas chitiniclastica is a kind of newfound Gram-negative conditioned pathogen which has caused some infection cases in six countrys.W.chitinclastica can cause sepsis and infection in some tissues of parasitifers including human being,Odocoileus virginianus,Delphinus delphis and Pangasius,which has already been found in China in 2008,but people know a little about it.In this study,W.chitiniclastica SH04 was used as the objective strain,and rounded analysis on physiological-biochemical characteristic and the genomic sequence of the strain were carried out for obtaining more data.Then a specificity detection method for W.chitiniclastica was established based on the comparative genomics research.In this study,the optimum medium components were determined for W.chitiniclastica as yeast extract 35 g/L,NaCl 10 g/L,and the optimum condition of culture,pH 6.7-7.0,temperature 37℃.Then,the physiological-biochemical characteristics of strain W.chitiniclastica SHO4 were analyzed and the results indicated that it was a knid of Gram-negative bacteria according to the BIOLOG GN and it can certainty utilize 24 carbon source,and 6 carbon source were uncertainty in 95 carbon source.It can generate oxidase,catalase,protease and chitinase.W.chitiniclastica SHO4 had strong resistance to vancomycin,clindamycin and benzene azole penicillin among 30 kinds of common antibiotics.Its whole-cell hydrolysate contained meso-DAP(meso-diaminopimelic acid)and PG,PE and PS were the main polar lipids in cell hydrolysate.There were 1971 CDS in the genomic sequence of strain W.chitiniclastica SHO4 and the length of the total genes is up to 1.88 Mb,the average length about 1002 bp.The average G+C mol%of the genes was 44.21%and the length of the total genes accounted for 98.27%of the length of the genome.By annotation,1601 CDS were predicted with biological function and there were a set of 16S/23S/5S rDNA operon,45 tRNA genes,50 tandem repeat sequences,48 transposon,18 IS and 2 CRISPR in the genome.Via performed comparative genomics research on two genomic sequence of W.chitiniclastica,we found that the two genome have well colinearity and there were 1786 CDS in the core genome.At last,we established a rapid real-time PCR methods to detect W.chitiniclastica and the method possessed better specificity and repeatability.19 CFU was available to be detected in 20 μL reaction system.