Genomics Study Of Recombinant Hansenula Anomala Ar_han0458 By Low Energy Ion Beam

Hansenula anomala has strong ester-producing and aroma-producing ability,strong fermentation and esterification ability under high temperature conditions.It is the main source of aroma in brewing industry,and is also a commonly used exogenous gene expression vector for ion implantation mediated transgenic studies.In this study,PacBio RSII sequencing technology was used to perform genomic de novo sequencing of the ion beam recombination Hansenula anomala Ar_Han0458 and its original strain As2340,and bioinformatics was used to study the genomics of Hansenula anomala before and after low-energy ion implantation from the aspects of genome profile,genome structure and components,gene function and phylogenetic evolution.The results of genome de novo sequencing showed that before low energy ion implantation,the genome size of As2340 was 13,760,069 bp,the length of Contig N50 was 1,883,332 bp and the GC content was 34.53%.1,897 tandem repeats with a total length of 238,164 bp were detected,interspersed repeats were 108,253 bp,and the repeat length accounted for 2.46% of the genome.932 non-coding RNA genes were predicted.In the 6,664 predicted coding genes,98.62% had homologous protein genes,and their genetic integrity reached 91.7%.The number of protein coding genes annotated to KOG,KEGG and GO databases were 4,391,2,119 and 3,580,respectively,and 3,937 GO functions were enriched.The results of de novo sequencing of Ar_Han0458,a Hansenula anomala recombination by low energy ion implantation,showed that its genome size was 13,646,815 bp and the length of Contig N50 was 1,346,975 bp,which were 113,254 bp and 536,357 bp less than the original strain respectively,while the GC content was the same as the original strain.In the genome of recombinant strain Ar_Han0458,1,874 tandem repeats with a total length of 227,825 bp were detected,and 107,439 bp of interspersed repeats.The length of repetitive sequences accounted for 2.52% of the genome.The number of predicted non-coding RNA genes and coding genes were 25 and 38 less than the original strain,respectively,while 44 GO function was missing.Using the genome of the original strain As2340 as the reference genome,the mutation of the recombinant strain Ar_Han0458 was analyzed by comparative genomics.The results showed that the number of SNP in the genome accounted for 0.15% of the genome,of which 34.50% occurred in the genome,17,779 loci were inserted or deleted,and the number of InDel in the gene was 5,997.It accounted for 33.69% of the total genome.The results of cluster analysis of gene families of Ar_Han0458 and its original strain As2340 and seven other yeasts by ion beam recombination showed that above 9 yeasts were clustered into 7,588 gene families,and 1,789 single copy homologous genes were used to construct phylogenetic tree.The results showed that the most closely related to the a Hansenula anomala was Wickerhamomyces anomalus,and the furthest was Schizosaccharomyces pombe.These findings enrich our understanding of the Hansenula anomala genome,deepen our comprehension of the mechanism of low-energy ion implantation-mediated microbial genomic variation,and also provided basic data for the subsequent studies on secondary metabolism regulation and genetic diversity of ion beam recombinant yeast.