Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory on Molecular Safety Assessment of Agri-GMO, MARA, Beijing, 100081, China.
BMC Plant Biol. 2024 Sep 4;24(1):829. doi: 10.1186/s12870-024-05548-w.
In research to improve the quality of transgenic crops, it is often necessary to introduce multiple functionally related genes into recipient plants simultaneously to improve crop genetic traits effectively. Compared with unidirectional promoters, bidirectional promoters simultaneously regulate the expression of multiple genes and improve the efficiency of biotechnology. Therefore, in this study, bidirectional gene pairs were systematically analyzed in Gossypium hirsutum TM-1, and the structure, function and evolutionary relationships of the bidirectional genes were analyzed. The endogenous bidirectional promoters of cotton were mined, and their specific regulatory elements and biological functions were explored to provide useful promoter resources and a theoretical basis for cultivating new cotton germplasms with excellent fiber quality.
Using an improved search model, a total of 1,383 bidirectional transcript pairs were identified in the Gossypium hirsutum TM-1 genome, and their gene structure and functional annotations were systematically analyzed. Thirty bidirectional intergenic sequences were randomly screened for promoter activity analysis via a transient expression system, and 25 intergenic sequences were found to have bidirectional promoter activity. Comparative analysis of the bidirectional gene profiles of the four cotton subspecies revealed that these subspecies presented abundant bidirectional gene pairs with high homology and that the bidirectional genes in the cotton subspecies were more similar in terms of their molecular functions, cellular components and biological processes. In addition, parallel analysis of bidirectional genes in dicotyledons and monocotyledons revealed that abundant bidirectional gene pairs exist in different species. Although the total number of orthologous bidirectional genes was similar, there was a significant difference in the number of orthologous bidirectional gene pairs between dicotyledons and monocotyledons. This evolutionary analysis of the function and structure of homologous bidirectional gene pairs in different varieties and different subspecies of the same species revealed potential pathways by which these gene pairs originated, which may be necessary for the evolution of a new species.
In this study, many bidirectional gene pairs in Gossypium hirsutum TM-1 were identified using computer programming, and systematic analysis was conducted to explore their functions and evolutionary relationships. In addition, the promoter activity of the bidirectional intergenic sequences was verified. The combination of computer programming screening, experimental validation and other methods is expected to provide preferred bidirectional promoters for transgenic breeding work via multigene cotransformation methods, and this information is valuable for genetic engineering research and applications.
在提高转基因作物质量的研究中，通常需要同时将多个功能相关的基因导入受体植物中，以有效改善作物遗传特性。与单向启动子相比，双向启动子可以同时调控多个基因的表达，提高生物技术的效率。因此，本研究在海岛棉 TM-1 中系统分析了双向基因对，分析了双向基因的结构、功能和进化关系。挖掘了棉花内源性双向启动子，探讨了其特异调控元件和生物学功能，为培育具有优良纤维品质的新型棉花种质资源提供了有用的启动子资源和理论依据。
利用改进的搜索模型，在海岛棉 TM-1 基因组中总共鉴定到 1383 个双向转录对，并对其基因结构和功能注释进行了系统分析。通过瞬时表达系统随机筛选了 30 个双向基因间序列进行启动子活性分析，发现 25 个基因间序列具有双向启动子活性。对四个棉亚种的双向基因图谱进行比较分析表明，这些亚种具有丰富的高同源性的双向基因对，棉亚种的双向基因在分子功能、细胞成分和生物学过程方面更为相似。此外，对双子叶植物和单子叶植物的双向基因进行平行分析表明，不同物种中存在丰富的双向基因对。虽然同源双向基因对的总数相似，但双子叶植物和单子叶植物之间的同源双向基因对的数量存在显著差异。对不同物种和同一物种不同亚种的同源双向基因对的功能和结构进行进化分析，揭示了这些基因对起源的潜在途径，这可能是新物种进化所必需的。
本研究利用计算机编程在海岛棉 TM-1 中鉴定到许多双向基因对，并进行了系统分析，以探讨其功能和进化关系。此外，验证了双向基因间序列的启动子活性。计算机编程筛选、实验验证等方法的结合，有望通过多基因共转化方法为转基因育种工作提供首选的双向启动子，为遗传工程研究和应用提供有价值的信息。
