Shivashakarappa Kuber, Marriboina Sureshbabu, Yadegari Zeinab, Paduri Vikas Reddy, Sachan Ritesh, Dumenyo Korsi, Taheri Ali
Department of Agricultural Science and Engineering, College of Agriculture, Tennessee State University, Nashville, TN, United States.
Department of Life and Physical Sciences, Fisk University, Nashville, TN, United States.
Front Chem. 2025 Mar 19;13:1542504. doi: 10.3389/fchem.2025.1542504. eCollection 2025.
Agriculture and food security face significant challenges due to population growth, climate change, and biotic and abiotic stresses. Enhancing crop productivity and quality through biotechnology is crucial in addressing these challenges. Genome engineering techniques, including gene cassette delivery into plant cells, aim to meet these demands. However, conventional biomolecule delivery methods have limitations such as poor efficacy, low regeneration capability, and potential cell damage. Nanoparticles, known for their success in drug delivery in animals, hold promise as DNA nanocarriers in plant sciences. This study explores the efficacy of carbon dots (CDs), synthesized rapidly and cost-effectively from citric acid monohydrate and β-alanine using a microwave-assisted method, as carriers for plasmid DNA delivery into plant tissues. The detailed characterization of carbon dots, evaluation of their binding ability with plasmid DNA, and phytotoxicity assessments were systematically conducted. The delivery and expression of plasmid DNA were successfully demonstrated in canola leaves via needleless syringe infiltration and in soybean root cells and protoplasts through passive diffusion. Additionally, the particle bombardment method facilitated the efficient delivery of plasmid DNA of varying sizes (4 kb, 11 kb, and 17 kb) into onion epidermal cells, as well as the successful delivery of plasmid DNA containing the GUS reporter gene into soybean embryos, using carbon dots as a binding agent between plasmid DNA and tungsten microcarrier. To our knowledge, this is the first study to report the use of carbon dots as a substitute for spermidine in such applications. Overall, our research presents a rapidly synthesized, cost-effective platform for efficient plasmid DNA delivery, establishing a foundation for using carbon dots as carriers for CRISPR and RNAi constructs in gene knockout and knockdown applications in plant tissues, with a comparison of their transformation efficiency against traditional delivery techniques.
由于人口增长、气候变化以及生物和非生物胁迫，农业和粮食安全面临重大挑战。通过生物技术提高作物生产力和品质对于应对这些挑战至关重要。基因组工程技术，包括将基因盒导入植物细胞，旨在满足这些需求。然而，传统的生物分子递送方法存在局限性，如效率低下、再生能力差以及潜在的细胞损伤。纳米颗粒在动物药物递送方面取得了成功，有望成为植物科学中的DNA纳米载体。本研究探索了通过微波辅助法由一水柠檬酸和β-丙氨酸快速且经济高效地合成的碳点作为质粒DNA递送至植物组织的载体的功效。系统地对碳点进行了详细表征、评估了它们与质粒DNA的结合能力以及进行了植物毒性评估。通过无针注射器浸润成功地在油菜叶片中证明了质粒DNA的递送和表达，并且通过被动扩散在大豆根细胞和原生质体中实现了递送和表达。此外，粒子轰击法利用碳点作为质粒DNA与钨微载体之间的结合剂，促进了不同大小（4 kb、11 kb和17 kb）的质粒DNA高效递送至洋葱表皮细胞，以及将含有GUS报告基因的质粒DNA成功递送至大豆胚胎中。据我们所知，这是第一项报道在这类应用中使用碳点替代亚精胺的研究。总体而言，我们的研究提出了一个快速合成、经济高效的高效质粒DNA递送平台，为在植物组织的基因敲除和敲低应用中使用碳点作为CRISPR和RNAi构建体的载体奠定了基础，并将其转化效率与传统递送技术进行了比较。
