Plant Biotechnology, Council of Scientific and Industrial Research-Central Institute of Medicinal and Aromatic Plants, Lucknow, 226015, India.
Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201002, India.
Protoplasma. 2021 Sep;258(5):1155-1162. doi: 10.1007/s00709-021-01631-3. Epub 2021 Mar 11.
Kalmegh [Andrographis paniculata (Burm.f.) Wall. ex Nees] is one of the most studied medicinal plants for pharmaceutical properties and phytochemistry. However, functional genomics studies in kalmegh are so far limited due to the unavailability of a robust tool for gene silencing. Here, we tested the application of virus-induced gene silencing (VIGS) in kalmegh using the well-known Tobacco rattle virus (TRV)-based vectors and achieved targeted silencing of phytoene desaturase (ApPDS) which is essential in plants for carotenoid biosynthesis that protects chlorophyll from photooxidation. ApPDS silencing in kalmegh leaves developed a typical photobleaching phenotype. The silencing of ApPDS was confirmed by analysing ApPDS transcript level and determining chlorophyll content in the leaves of VIGS seedlings. The analysis revealed ~30% reduction in chlorophyll content, and 40 to 60% reduction in ApPDS transcript level in the leaves of VIGS seedlings. These findings clearly demonstrated the applicability of VIGS in kalmegh using TRV-based vectors. The VIGS protocol presented in this study might be useful for studying gene function related to medicinal and agricultural traits in kalmegh.
穿心莲[穿心莲（Burm.f.）Wall. ex Nees]是研究最多的药用植物之一，具有药物特性和植物化学特性。然而，由于缺乏有效的基因沉默工具，穿心莲的功能基因组学研究迄今为止还很有限。在这里，我们使用众所周知的烟草脆裂病毒（TRV）载体测试了病毒诱导的基因沉默（VIGS）在穿心莲中的应用，并实现了对类胡萝卜素生物合成中必不可少的脱植烯酶（ApPDS）的靶向沉默，该酶可保护叶绿素免受光氧化。穿心莲叶片中的 ApPDS 沉默导致了典型的光漂白表型。通过分析 VIGS 幼苗叶片中的 ApPDS 转录本水平和测定叶绿素含量，证实了 ApPDS 的沉默。分析表明，VIGS 幼苗叶片中的叶绿素含量降低了约 30%，ApPDS 转录本水平降低了 40%至 60%。这些发现清楚地表明，使用基于 TRV 的载体，VIGS 可应用于穿心莲。本研究中提出的 VIGS 方案可能有助于研究与穿心莲药用和农业特性相关的基因功能。
