College of Food and Biochemical Engineering, Guangxi Science and Technology Normal University, Guangxi Laibin, 546199, People's Republic of China.
Key Laboratory for Zhuang and Yao Pharmaceutical Quality Biology, Guangxi Science and Technology Normal University, Laibin, 546199, People's Republic of China.
Plant Cell Rep. 2024 Feb 22;43(3):77. doi: 10.1007/s00299-023-03126-2.
We reported the mitochondrial genome of Ventilago leiocarpa for the first time. Two and one sites lead to the generation of stop and stat codon through editing were verified. Ventilago leiocarpa, a member of the Rhamnaceae family, is frequently utilized in traditional medicine due to the medicinal properties of its roots. In this study, we successfully assembled the mitogenome of V. leiocarpa using both BGI short reads and Nanopore long reads. This mitogenome has a total length of 331,839 bp. The annotated results showed 36 unique protein-coding, 16 tRNA and 3 rRNA genes in this mitogenome. Furthermore, we confirmed the presence of a branched structure through the utilization of long reads mapping, PCR amplification, and Sanger sequencing. Specifically, the ctg1 can form a single circular molecule or combine with ctg4 to form a linear molecule. Likewise, ctg2 can form a single circular molecule or can be connected to ctg4 to form a linear molecule. Subsequently, through a comparative analysis of the mitogenome and cpgenome sequences, we identified ten mitochondrial plastid sequences (MTPTs), including two complete protein-coding genes and five complete tRNA genes. The existence of MTPTs was verified by long reads. Colinear analysis showed that the mitogenomes of Rosales were highly divergent in structure. Finally, we identified 545 RNA editing sites involving 36 protein-coding genes by Deepred-mt. To validate our findings, we conducted PCR amplification and Sanger sequencing, which confirmed the generation of stop codons in atp9-223 and rps10-391, as well as the generation of a start codon in nad4L-2. This project reported the complex structure and RNA editing event of the V. Leiocarpa mitogenome, which will provide valuable information for the study of mitochondrial gene expression.
我们首次报道了榕属(Ventilago leiocarpa)的线粒体基因组。通过编辑验证了两个和一个导致产生终止和起始密码子的位点。榕属(Rhamnaceae 科)的一员,由于其根部的药用特性,经常被用于传统医学。在这项研究中,我们使用 BGI 短读和纳米孔长读成功组装了 V. leiocarpa 的线粒体基因组。这个线粒体基因组总长为 331839bp。注释结果表明,这个线粒体基因组中有 36 个独特的蛋白质编码基因、16 个 tRNA 和 3 个 rRNA 基因。此外,我们通过利用长读测序、PCR 扩增和 Sanger 测序证实了存在分支结构。具体来说,ctg1 可以形成一个单一的圆形分子或与 ctg4 结合形成一个线性分子。同样,ctg2 可以形成一个单一的圆形分子,也可以与 ctg4 连接形成一个线性分子。随后,通过对线粒体基因组和 cpgenome 序列的比较分析,我们鉴定了 10 个线粒体质体序列(MTPTs),包括两个完整的蛋白质编码基因和五个完整的 tRNA 基因。MTPTs 的存在通过长读验证。共线性分析表明,蔷薇科的线粒体基因组在结构上高度分化。最后,我们通过 Deepred-mt 鉴定了 545 个涉及 36 个蛋白质编码基因的 RNA 编辑位点。为了验证我们的发现,我们进行了 PCR 扩增和 Sanger 测序,证实了在 atp9-223 和 rps10-391 产生终止密码子,以及在 nad4L-2 产生起始密码子。本项目报道了榕属线粒体基因组的复杂结构和 RNA 编辑事件,为线粒体基因表达研究提供了有价值的信息。