Santana Israel, Hu Peiguang, Jeon Su-Ji, Castillo Chris, Tu Hann, Giraldo Juan Pablo
Department of Botany and Plant Sciences, University of California, Riverside, CA 92521, USA.
Bio Protoc. 2021 Jun 20;11(12):e4060. doi: 10.21769/BioProtoc.4060.
Plant nanobiotechnology is a flourishing field that uses nanomaterials to study and engineer plant function. Applications of nanotechnology in plants have great potential as tools for improving crop yield, tolerance to disease and environmental stress, agrochemical delivery of pesticides and fertilizers, and genetic modification and transformation of crop plants. Previous studies have used nanomaterials functionalized with chemicals, including biocompatible polymers with charged, neutral, or hydrophobic functional groups, to improve nanomaterial uptake and localization in plant cells. Recently, the use of biorecognition motifs such as peptides has been demonstrated to enable the targeted delivery of nanoparticles in plants ( Santana , 2020 ). Herein, we describe a bio-protocol to target nanoparticles with chemical cargoes to chloroplasts in plant leaves and assess targeting efficiency using advanced analytical tools, including confocal microscopy and elemental analysis. We also describe the use of isothermal titration calorimetry to determine the affinity of nanomaterials for their chemical cargoes. Nanotechnology-based methods for targeted delivery guided by conserved plant molecular recognition mechanisms will provide more robust plant bioengineering tools across diverse plant species. Graphic abstract: Targeted delivery of nanomaterials with chemical cargoes to chloroplasts enabled by plant biorecognition.
植物纳米生物技术是一个蓬勃发展的领域,它利用纳米材料来研究和设计植物功能。纳米技术在植物中的应用作为提高作物产量、抗病和抗环境胁迫能力、农药和化肥的农用化学品递送以及作物植物的基因修饰和转化的工具具有巨大潜力。先前的研究使用了用化学物质功能化的纳米材料,包括带有带电、中性或疏水官能团的生物相容性聚合物,以提高纳米材料在植物细胞中的摄取和定位。最近,已经证明使用肽等生物识别基序能够在植物中实现纳米颗粒的靶向递送(桑塔纳,2020年)。在此,我们描述了一种生物协议,用于将带有化学货物的纳米颗粒靶向植物叶片中的叶绿体,并使用先进的分析工具(包括共聚焦显微镜和元素分析)评估靶向效率。我们还描述了使用等温滴定量热法来确定纳米材料与其化学货物的亲和力。基于保守植物分子识别机制的靶向递送纳米技术方法将为不同植物物种提供更强大的植物生物工程工具。图形摘要:通过植物生物识别实现将带有化学货物的纳米材料靶向递送至叶绿体。