Shahid Muhammad Shafiq, Sattar Muhammad Naeem, Iqbal Zafar, Raza Amir, Al-Sadi Abdullah M
Department of Plant Sciences, College of Agricultural and Marine Sciences, Sultan Qaboos University, Muscat, Oman.
Central Laboratories, King Faisal University, Hofuf, Saudi Arabia.
Front Microbiol. 2021 Jan 12;11:609376. doi: 10.3389/fmicb.2020.609376. eCollection 2020.
In recent years, next-generation sequencing (NGS) and contemporary Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-CRISPR-associated (Cas) technologies have revolutionized the life sciences and the field of plant virology. Both these technologies offer an unparalleled platform for sequencing and deciphering viral metagenomes promptly. Over the past two decades, NGS technologies have improved enormously and have impacted plant virology. NGS has enabled the detection of plant viruses that were previously undetectable by conventional approaches, such as quarantine and archeological plant samples, and has helped to track the evolutionary footprints of viral pathogens. The CRISPR-Cas-based genome editing (GE) and detection techniques have enabled the development of effective approaches to virus resistance. Different versions of CRISPR-Cas have been employed to successfully confer resistance against diverse plant viruses by directly targeting the virus genome or indirectly editing certain host susceptibility factors. Applications of CRISPR-Cas systems include targeted insertion and/or deletion, site-directed mutagenesis, induction/expression/repression of the gene(s), epigenome re-modeling, and SNPs detection. The CRISPR-Cas toolbox has been equipped with precision GE tools to engineer the target genome with and without double-stranded (ds) breaks or donor templates. This technique has also enabled the generation of transgene-free genetically engineered plants, DNA repair, base substitution, prime editing, detection of small molecules, and biosensing in plant virology. This review discusses the utilities, advantages, applications, bottlenecks of NGS, and CRISPR-Cas in plant virology.
近年来，新一代测序（NGS）技术和当代成簇规律间隔短回文重复序列（CRISPR）-CRISPR相关蛋白（Cas）技术彻底改变了生命科学以及植物病毒学领域。这两种技术都为快速测序和解读病毒宏基因组提供了无与伦比的平台。在过去二十年中，NGS技术有了巨大改进，并对植物病毒学产生了影响。NGS能够检测出传统方法无法检测到的植物病毒，如检疫和考古植物样本中的病毒，并有助于追踪病毒病原体的进化足迹。基于CRISPR-Cas的基因组编辑（GE）和检测技术推动了有效的抗病毒方法的发展。不同版本的CRISPR-Cas已被用于通过直接靶向病毒基因组或间接编辑某些宿主易感性因子，成功赋予对多种植物病毒的抗性。CRISPR-Cas系统的应用包括靶向插入和/或缺失、定点诱变、基因诱导/表达/抑制、表观基因组重塑以及单核苷酸多态性（SNP）检测。CRISPR-Cas工具箱配备了精确的GE工具，可在有或无双链（ds）断裂或供体模板的情况下对目标基因组进行工程改造。这项技术还能够产生无转基因的基因工程植物、进行DNA修复、碱基替换、碱基编辑、小分子检测以及植物病毒学中的生物传感。本综述讨论了NGS和CRISPR-Cas在植物病毒学中的用途、优势、应用及瓶颈。
