Institute of Biology, Leiden University, Leiden, The Netherlands.
Microbiologyopen. 2021 Mar;10(2):e1180. doi: 10.1002/mbo3.1180.
We report here the complete genome sequence of the Rhizobium rhizogenes (formerly Agrobacterium rhizogenes) strain LBA9402 (NCPPB1855rifR), a pathogenic strain causing hairy root disease. To assemble a complete genome, we obtained short reads from Illumina sequencing and long reads from Oxford Nanopore Technology sequencing. The genome consists of a 3,958,212 bp chromosome, a 2,005,144 bp chromid (secondary chromosome) and a 252,168 bp Ri plasmid (pRi1855), respectively. The primary chromosome was very similar to that of the avirulent biocontrol strain K84, but the chromid showed a 724 kbp deletion accompanied by a large 1.8 Mbp inversion revealing the dynamic nature of these secondary chromosomes. The sequence of the agropine Ri plasmid was compared to other types of Ri and Ti plasmids. Thus, we identified the genes responsible for agropine catabolism, but also a unique segment adjacent to the TL region that has the signature of a new opine catabolic gene cluster including the three genes that encode the three subunits of an opine dehydrogenase. Our sequence analysis also revealed a novel gene at the very right end of the TL-DNA, which is unique for the agropine Ri plasmid. The protein encoded by this gene was most related to the succinamopine synthases of chrysopine and agropine Ti plasmids and thus may be involved in the synthesis of the unknown opine that can be degraded by the adjacent catabolic cluster. The available sequence will facilitate the use of R. rhizogenes and especially LBA9402 in both the laboratory and for biotechnological purposes.
我们在此报告根瘤农杆菌(以前称为根癌农杆菌)菌株 LBA9402(NCPPB1855rifR)的完整基因组序列,该菌株是引起毛状根病的致病菌株。为了组装完整的基因组,我们从 Illumina 测序中获得了短读序列,从 Oxford Nanopore Technology 测序中获得了长读序列。该基因组由一个 3958212bp 的染色体、一个 2005144bp 的染色质(次级染色体)和一个 252168bp 的 Ri 质粒(pRi1855)组成。主染色体与无毒生物防治菌株 K84 的染色体非常相似,但染色质显示出 724kbp 的缺失,同时伴随着一个 1.8Mbp 的倒位,揭示了这些次级染色体的动态性质。农杆菌 Ri 质粒的序列与其他类型的 Ri 和 Ti 质粒进行了比较。因此,我们确定了负责农杆碱分解的基因,还鉴定了与 TL 区相邻的独特片段,该片段具有新的(opine)分解代谢基因簇的特征,包括编码三个亚基的三个基因三丁胺脱氢酶。我们的序列分析还揭示了 TL-DNA 非常右端的一个新基因,该基因是农杆碱 Ri 质粒所特有的。该基因编码的蛋白质与 chrysopine 和 agropine Ti 质粒的琥珀酸酰基三丁胺合成酶最相关,因此可能参与了未知 opine 的合成,该 opine 可被相邻的分解代谢簇降解。现有的序列将促进根瘤农杆菌,特别是 LBA9402 在实验室和生物技术方面的应用。