Dipartimento di Scienze Agrarie e Forestali, Università degli Studi della Tuscia, Via S. Camillo de Lellis snc, Viterbo, 01100, Italy.
CIHEAM-Mediterranean Agronomic Institute of Bari, Via Ceglie 9, Valenzano, 70010, Italy.
Environ Microbiol Rep. 2022 Apr;14(2):274-285. doi: 10.1111/1758-2229.13048. Epub 2022 Feb 2.
Pseudomonas savastanoi pv. savastanoi, the causal agent of olive knot disease, is a fluorescent Gram-negative bacterium classified, according to the specific LOPAT profile, as Ib. However, during the 90s, a number of atypical non-fluorescent levan-positive strains of Pseudomonas savastanoi pv. savastanoi have been unexpectedly isolated from olive knots in Central Italy. Since its first report, several studies were conducted on this species variant, but its genome sequence has never been reported. The complete genome sequence and two additional plasmids of PVFi1, a representative strain, were here obtained using a hybrid sequencing approach with both Oxford Nanopore Technology and Illumina sequencing. A thorough genomic analysis unravelled several genetic features of this peculiar strain, showing a transposase insertion downstream a fragmented copy of the levansucrase gene. The same features were previously reported on levan-negative Pseudomonas savastanoi pv. savastanoi strains. In addition, a second copy of the levansucrase gene fully equipped for a gene expression and comparable to the levan-positive Pseudomonas savastanoi pv. glycinea, may explain the levan-positive test. This result provides a solid genetic demonstration that the bacterial species Pseudomonas savastanoi contains either levan-positive or levan-negative strains, providing insights for an update of the related LOPAT classification.
橄榄瘿多胞菌 pv. savastanoi,引起橄榄瘤病的病原体,是一种荧光革兰氏阴性细菌,根据特定的 LOPAT 图谱,被归类为 Ib。然而,在 90 年代,一些非荧光的蔗聚糖阳性的不典型橄榄瘿多胞菌 pv. savastanoi 菌株出乎意料地从意大利中部的橄榄瘤中分离出来。自首次报道以来,对这种变异株进行了多项研究,但从未报道过其基因组序列。本研究采用 Oxford Nanopore Technology 和 Illumina 测序的混合测序方法,获得了代表性菌株 PVFi1 的完整基因组序列和两个额外的质粒。对该特殊菌株的深入基因组分析揭示了其一些遗传特征,表明在蔗糖酶基因的不完整拷贝下游有一个转座酶插入。以前在蔗聚糖阴性的橄榄瘿多胞菌 pv. savastanoi 菌株中也报道过类似的特征。此外,蔗糖酶基因的第二个完全具备基因表达能力的拷贝,与蔗聚糖阳性的大豆橄榄瘿多胞菌 pv. glycinea 相当,可能解释了该菌株的蔗聚糖阳性测试。这一结果为细菌物种橄榄瘿多胞菌包含蔗聚糖阳性或蔗聚糖阴性菌株提供了可靠的遗传证据,为相关 LOPAT 分类的更新提供了依据。