Characterization of the Mitochondrial Genome of subsp. Rousi Based on High-Throughput Sequencing and Elucidation of Its Evolutionary Mechanisms.

ssp. Rousi a species of significant ecological and economic value that is native to the Qinghai-Tibet Plateau and arid/semi-arid regions. Investigating the mitochondrial genome can elucidate stress adaptation mechanisms, population genetic structure, and hybrid evolutionary history, offering molecular insights for ecological restoration and species conservation. However, the genetic information and evolutionary mechanisms of its mitochondrial genome remain poorly understood. This study aimed to assemble the complete mitochondrial genome of L. ssp. using Illumina sequencing, uncovering its structural features, evolutionary pressures, and environmental adaptability and addressing the research gap regarding mitochondrial genomes within the genus. The study assembled a 454,444 bp circular mitochondrial genome of ssp. , with a GC content of 44.86%. A total of 73 genes and 3 pseudogenes were annotated, with the notable absence of the gene, which is present in related species. The genome exhibits significant codon usage bias, particularly with high-frequency use of the alanine codon GCU and the isoleucine codon AUU. Additionally, 449 repetitive sequences, potentially driving genome recombination, were identified. Our evolutionary pressure analysis revealed that most genes are under purifying selection, while genes such as and exhibit positive selection. A nucleotide diversity analysis revealed that the gene exhibits the highest variation, whereas is the most conserved. Meanwhile, phylogenetic analysis showed that ssp. from China is most closely related to , with extensive homologous sequences (49.72% of the chloroplast genome) being identified between the chloroplast and mitochondrial genomes, indicating active inter-organellar gene transfer. Furthermore, 539 RNA editing sites, primarily involving hydrophilic-to-hydrophobic amino acid conversions, were predicted, potentially regulating mitochondrial protein function. Our findings establish a foundation for genetic improvement and research on adaptive evolutionary mechanisms in the genus, offering a novel case study for plant mitochondrial genome evolution theory.