Establishing a Virus-Induced Gene Silencing System in <i>Lycoris chinensis</i>
2023
Guanghao Cheng | Xiaochun Shu | Zhong Wang | Ning Wang | Fengjiao Zhang
<i>Lycoris</i> is an important plant with both medicinal and ornamental values. However, it does not have an efficient genetic transformation system, which makes it difficult to study gene function of the genus. Virus-induced gene silencing (VIGS) is an effective technique for studying gene functions in plants. In this study, we develop an efficient virus-induced gene-silencing (VIGS) system using the leaf tip needle injection method. The widely used TRV vector is constructed, and the <i>Cloroplastos Alterados 1</i> (<i>CLA1</i>) and <i>Phytoene Desaturase</i> (<i>PDS</i>) genes are selected as visual indicators in the VIGS system. As a result, it is observed that leaves infected with TRV-<i>LcCLA1</i> and TRV-<i>LcPDS</i> both show a yellowing phenotype (loss of green), and the chlorosis range of TRV-<i>LcCLA1</i> was larger and deeper than that of TRV-<i>LcPDS</i>. qRT-PCR results show that the expression levels of <i>LcCLA1</i> and <i>LcPDS</i> are significantly reduced, and the silencing efficiency of <i>LcCLA1</i> is higher than that of <i>LcPDS</i>. These results indicate that the VIGS system of <i>L. chinensis</i> was preliminarily established, and <i>LcCLA1</i> is more suitable as a gene-silencing indicator. For the monocotyledonous plant leaves with a waxy surface, the leaf tip injection method greatly improves the infiltration efficiency. The newly established VIGS system will contribute to gene functional research in <i>Lycoris</i> species.
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