Division of Applied Life Science (BK21 FOUR Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju 660-701, Korea.
Department of Horticultural Bioscience, Pusan National University, Miryang 50463, Korea.
Int J Mol Sci. 2021 Feb 13;22(4):1878. doi: 10.3390/ijms22041878.
Tomato is one of the major vegetable crops consumed worldwide. (TYLCV) and fungal sp. are devastating pathogens causing yellow leaf curl disease and powdery mildew. Such viral and fungal pathogens reduce tomato crop yields and cause substantial economic losses every year. Several commercial tomato varieties include () and () locus that carries the susceptibility (-gene) factors for TYLCV and powdery mildew, respectively. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) is a valuable genome editing tool to develop disease-resistant crop varieties. In this regard, targeting susceptibility factors encoded by the host plant genome instead of the viral genome is a promising approach to achieve pathogen resistance without the need for stable inheritance of CRISPR components. In this study, the CRISPR/Cas9 system was employed to target the and for trait introgression in elite tomato cultivar BN-86 to confer host-mediated immunity against pathogens. -knockout lines were successfully generated, carrying the biallelic indel mutations. The pathogen resistance assays in mutant lines confirmed the suppressed accumulation of TYLCV and restricted the spread to non-inoculated plant parts. Generated knockout lines for the showed complete resistance to powdery mildew fungus. Overall, our results demonstrate the efficiency of the CRISPR/Cas9 system to introduce targeted mutagenesis for the rapid development of pathogen-resistant varieties in tomato.
番茄是全球消费的主要蔬菜作物之一。（TYLCV）和真菌 sp. 是毁灭性的病原体，导致黄卷叶病和白粉病。这些病毒和真菌病原体每年都会降低番茄作物的产量并造成巨大的经济损失。几种商业番茄品种包括携带对 TYLCV 和白粉病敏感（-基因）因素的 () 和 () 基因座。成簇规律间隔短回文重复 (CRISPR)/CRISPR 相关蛋白 (Cas) 是一种有价值的基因组编辑工具，可用于开发抗病作物品种。在这方面，针对宿主植物基因组编码的易感性因素而不是病毒基因组是一种有前途的方法，可以在不需要 CRISPR 成分稳定遗传的情况下实现病原体抗性。在这项研究中，CRISPR/Cas9 系统被用于靶向 BN-86 优良番茄品种中的 和 ，以赋予宿主对病原体的介导免疫。成功生成了 -敲除系，携带双等位基因缺失突变。在 -突变系中的病原体抗性测定证实了 TYLCV 的积累受到抑制，并限制了其在未接种植物部分的传播。生成的 敲除系对白粉病真菌表现出完全抗性。总的来说，我们的结果表明 CRISPR/Cas9 系统能够有效地进行靶向诱变，从而快速开发番茄的抗病品种。
