An Avirulent Ralstonia Solanacearum Strain Undergoes Phenotype Conversion from a Pathogenic Strain Under Natural Environment

De-ju Chen, Haifeng Zhang, Yanli Li, Yanping Chen, Xuefang Zheng, Jieping Wang, Jiamei Che, Bo Liu
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Abstract

An avirulent R. solanacearum strain named FJAT-1458 was isolated from living tomato vessel and it showed no toxicity to tomato, pepper and eggplant. Multilocus sequence analysis (MLSA) based on eight genes (egl, hrpB, mutS, pehA, recA, rpoA, rpoB and rpoC) and whole genome average nucleotide identity (ANI) analysis suggested that strain FJAT-1458 belong to phylotype I. Genome sequence of the strain FJAT-1458 revealed a circular chromosome and a circular megaplasmid with whole genome size of 6,059,899 bp and GC content of 66.78%. Functional annotation of FJAT-1458 showed a total of 5,442 genes, with 5,166 protein-encoding genes, 202 pseudogenes and 74 noncoding RNA genes. Among which, 3,938 protein-coding genes can be assigned to 23 COG families, and 1,521 of them had KEGG orthologs. Prophage prediction using PHASTER revealed 12 prophages, including 7 intact, 1 questionable and 4 incomplete prophages. Comparative genome analyses between GMI1000 and FJAT-1458 showed that most of the virulence factors were well conserved and only small portion of them were distinct between them. Two genes, including a methyltransferase and an ISL3 family transposase genes, were identified to be inserted immediately upstream (141 bp) of phcA gene, which assumed to be responsible for avirulence of strain FJAT-1458. It is suggested that strain FJAT-1458 was originated from a wild-type pathogenic strain through an accident phenotype conversion, which is like those when cultured under experimental conditions. Our study provides new insight into the evolution of virulence in R. solanacearum strain under natural environment.
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在自然环境下,一株无毒的茄青枯菌由致病菌株进行表型转化
从活番茄容器中分离到一株无毒性番茄红僵菌FJAT-1458,该菌株对番茄、辣椒和茄子均无毒性。基于8个基因(egl、hrpB、mutS、pehA、recA、rpoA、rpoB和rpoC)的多位点序列分析(MLSA)和全基因组平均核苷酸同源性(ANI)分析表明,菌株FJAT-1458属于ⅰ种型。菌株基因组序列显示环状染色体和环状巨质粒,全基因组大小为6059,899 bp, GC含量为66.78%。FJAT-1458的功能注释共显示5442个基因,其中蛋白编码基因5166个,假基因202个,非编码RNA基因74个。其中,3938个蛋白编码基因可归属于23个COG家族,其中1521个具有KEGG同源基因。利用PHASTER进行噬菌体预测,共发现12个噬菌体,其中完整的7个,可疑的1个,不完整的4个。GMI1000与FJAT-1458的基因组比较分析表明,大部分毒力因子保守性较好,只有一小部分毒力因子存在差异。两个基因,包括一个甲基转移酶和一个ISL3家族转座酶基因,被发现插入phcA基因的上游(141 bp),这被认为是菌株FJAT-1458无毒的原因。推测菌株FJAT-1458是由野生型致病菌株通过意外表型转化而产生的,与在实验条件下培养时的表型转化相同。本研究为自然环境下茄青霉毒力的进化提供了新的认识。
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