She Linwei, Cheng Xuejiao, Jiang Peng, Shen Simin, Dai Fangxiu, Run Yonghang, Zhu Mengting, Tavakoli Mahmoud, Yang Xueming, Wang Xiu-E, Xiao Jin, Chen Caiyan, Kang Zhenhui, Huang Jian, Zhang Wenli
State Key Laboratory of Crop Genetics & Germplasm Enhancement and Utilization, CIC-MCP, Nanjing Agricultural University, Nanjing, Jiangsu, China.
School of Biology & Basic Medical Science, Soochow University, Suzhou, Jiangsu, China.
Plant Biotechnol J. 2025 Apr;23(4):1139-1152. doi: 10.1111/pbi.14573. Epub 2025 Feb 19.
Genotype restriction poses a significant bottleneck to stable transformation in the vast majority of plant species, thereby severely impeding advancement in plant bioengineering, particularly for crops. Nanoparticles (NPs) can serve as effective carriers for the transient delivery of nucleic acids, facilitating gene overexpression or silencing in plants in a genotype-independent manner. However, the applications of NP-mediated transient systems in comprehensive genomic studies remained underexplored in plants, especially in crops that face challenges in genetic transformation. Consequently, there is an urgent need for efficient NP-mediated delivery systems capable of generating whole plants or seedlings with uniformly transformed nucleic acids. We have developed a straightforward and efficient modified carbon dot (MCD)-mediated transient transformation system for delivering DNA plasmids into the seeds of wheat, which is also applicable to other plant species. This system facilitates the generation of whole seedlings that contain the transferred DNA plasmids. Furthermore, our study demonstrates that this system serves as an excellent platform for conducting functional genomic studies in wheat, including the validation of gene functions, protein interactions and regulation, omics studies, and genome editing. This advancement significantly enhances functional genomic research for any plants or crops that face challenges in stable transformation. Thus, our study provides for the first time evidence of new applications for MCDs in functional genomics and epigenomic studies, and bioengineering potentially leading to the improvement of desirable agronomic traits in crops.
基因型限制在绝大多数植物物种的稳定转化中构成了重大瓶颈，从而严重阻碍了植物生物工程的进展，尤其是对农作物而言。纳米颗粒（NPs）可作为核酸瞬时递送的有效载体，以不依赖基因型的方式促进植物中的基因过表达或沉默。然而，NP介导的瞬时系统在植物全面基因组研究中的应用，尤其是在面临遗传转化挑战的农作物中，仍未得到充分探索。因此，迫切需要能够产生具有均匀转化核酸的完整植株或幼苗的高效NP介导递送系统。我们开发了一种简单高效的修饰碳点（MCD）介导的瞬时转化系统，用于将DNA质粒递送至小麦种子中，该系统也适用于其他植物物种。该系统有助于生成含有转移DNA质粒的完整幼苗。此外，我们的研究表明，该系统是在小麦中进行功能基因组学研究的优秀平台，包括基因功能验证、蛋白质相互作用与调控、组学研究以及基因组编辑。这一进展显著增强了对任何在稳定转化方面面临挑战的植物或作物的功能基因组学研究。因此，我们的研究首次提供了MCD在功能基因组学和表观基因组学研究以及生物工程中的新应用证据，这些应用可能会改善作物中理想的农艺性状。
