Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD 4072, Australia.
School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD 4072, Australia.
Plant Physiol. 2021 Oct 5;187(2):886-899. doi: 10.1093/plphys/kiab303.
Topical application of double-stranded RNA (dsRNA) can induce RNA interference (RNAi) and modify traits in plants without genetic modification. However, delivering dsRNA into plant cells remains challenging. Using developing tomato (Solanum lycopersicum) pollen as a model plant cell system, we demonstrate that layered double hydroxide (LDH) nanoparticles up to 50 nm in diameter are readily internalized, particularly by early bicellular pollen, in both energy-dependent and energy-independent manners and without physical or chemical aids. More importantly, these LDH nanoparticles efficiently deliver dsRNA into tomato pollen within 2-4 h of incubation, resulting in an 89% decrease in transgene reporter mRNA levels in early bicellular pollen 3-d post-treatment, compared with a 37% decrease induced by the same dose of naked dsRNA. The target gene silencing is dependent on the LDH particle size, the dsRNA dose, the LDH-dsRNA complexing ratio, and the treatment time. Our findings indicate that LDH nanoparticles are an effective nonviral vector for the effective delivery of dsRNA and other biomolecules into plant cells.
双层氢氧化物（LDH）纳米颗粒可高效递送入花粉细胞并诱导基因沉默
双层氢氧化物（LDH）纳米颗粒作为一种非病毒载体，可将双链 RNA（dsRNA）及其它生物分子递送入植物细胞
摘要：将双链 RNA（dsRNA）进行局部施用能够在不进行基因修饰的情况下诱导 RNA 干扰（RNAi）并改变植物的特性。然而，将 dsRNA 递送入植物细胞仍然具有挑战性。本研究以发育中的番茄（Solanum lycopersicum）花粉作为植物细胞模型系统，证明了直径高达 50nm 的层状双氢氧化物（LDH）纳米颗粒能够以能量依赖和非依赖的方式被轻易地内吞，尤其是早期的二细胞花粉，而无需物理或化学辅助。更为重要的是，这些 LDH 纳米颗粒能够在孵育 2-4 小时内将 dsRNA 高效递送入番茄花粉中，与相同剂量的裸露 dsRNA 相比，处理 3 天后早期二细胞花粉中转基因报告 mRNA 水平降低了 89%。靶基因沉默依赖于 LDH 纳米颗粒的粒径、dsRNA 的剂量、LDH-dsRNA 复合物的比例和处理时间。我们的研究结果表明，LDH 纳米颗粒是一种有效的非病毒载体，可将 dsRNA 和其它生物分子有效递送入植物细胞。
