Sanya Institute of China Agricultural University, Sanya, Hainan, 572025, China.
Department of Plant Pathology, Beijing Key Laboratory of Seed Disease Testing and Control, MOA Key Lab of Pest Monitoring and Green Management, China Agricultural University, Beijing, 100193, China.
BMC Plant Biol. 2024 Jul 8;24(1):645. doi: 10.1186/s12870-024-05319-7.
Melia azedarach is a species of enormous value of pharmaceutical industries. Although the chloroplast genome of M. azedarach has been explored, the information of mitochondrial genome (Mt genome) remains surprisingly limited. In this study, we used a hybrid assembly strategy of BGI short-reads and Nanopore long-reads to assemble the Mt genome of M. azedarach. The Mt genome of M. azedarach is characterized by two circular chromosomes with 350,142 bp and 290,387 bp in length, respectively, which encodes 35 protein-coding genes (PCGs), 23 tRNA genes, and 3 rRNA genes. A pair of direct repeats (R1 and R2) were associated with genome recombination, resulting in two conformations based on the Sanger sequencing and Oxford Nanopore sequencing. Comparative analysis identified 19 homologous fragments between Mt and chloroplast genome, with the longest fragment of 12,142 bp. The phylogenetic analysis based on PCGs were consist with the latest classification of the Angiosperm Phylogeny Group. Notably, a total of 356 potential RNA editing sites were predicted based on 35 PCGs, and the editing events lead to the formation of the stop codon in the rps10 gene and the start codons in the nad4L and atp9 genes, which were verified by PCR amplification and Sanger sequencing. Taken together, the exploration of M. azedarach gap-free Mt genome provides a new insight into the evolution research and complex mitogenome architecture.
苦楝是一种具有巨大药用价值的物种。尽管已经对苦楝的叶绿体基因组进行了探索,但线粒体基因组(Mt 基因组)的信息仍然非常有限。在本研究中,我们使用了 BGI 短读和 Nanopore 长读的混合组装策略来组装苦楝的 Mt 基因组。苦楝的 Mt 基因组由两个圆形染色体组成,分别长 350142bp 和 290387bp,编码 35 个蛋白编码基因(PCGs)、23 个 tRNA 基因和 3 个 rRNA 基因。一对直接重复(R1 和 R2)与基因组重组相关,导致基于 Sanger 测序和 Oxford Nanopore 测序的两种构象。比较分析确定了 Mt 和叶绿体基因组之间的 19 个同源片段,最长片段为 12142bp。基于 PCGs 的系统发育分析与被子植物系统发育组的最新分类一致。值得注意的是,基于 35 个 PCGs 预测了总共 356 个潜在的 RNA 编辑位点,编辑事件导致 rps10 基因中的终止密码子和 nad4L 和 atp9 基因中的起始密码子的形成,通过 PCR 扩增和 Sanger 测序验证了这些编辑事件。总之,对无间隙苦楝 Mt 基因组的探索为进化研究和复杂的线粒体基因组结构提供了新的见解。