Feng Shan, Chen Jin-Da, Bao Ai-Ke
State Key Laboratory of Herbage Improvement and Grassland Agro-Ecosystems, Key Laboratory of Grassland Livestock Industry Innovation, Ministry of Agriculture and Rural Affairs, College of Pastoral Agriculture Science and Technology, Lanzhou University, Lanzhou, 730000, China.
Plant Methods. 2025 Aug 7;21(1):107. doi: 10.1186/s13007-025-01427-z.
, a halophytic model plant exhibiting remarkable tolerance to abiotic stress, possesses valuable genetic resources for crop improvement. However, the absence of a reliable and stable transformation systems has impeded the functional characterization of candidate genes in this species. In this study, we established an efficient virus-induced gene silencing (VIGS) system in by constructing a tobacco rattle virus (TRV)-based vector targeting the phytoene desaturase gene (), whose silencing induces an easily recognizable photobleaching phenotype. Through comparative analyses of inoculation materials and methods for VIGS implementation in , we found that inoculating germinated seeds with suspension (OD = 0.8) carrying TRV2: vectors through vacuum-assisted agroinfiltration (0.5 kPa, 10 min) further achieving an average silencing efficiency of approximately 16.4%. Systemic photobleaching phenotypes appeared in newly emerged leaves at about 15 days post-inoculation, accompanied by a 40–80% reduction in transcript abundance, as confirmed by qRT-PCR. Furthermore, we successfully validated this VIGS system by silencing two aquaporin genes ( and ), achieving 60.3–69.5% knockdown efficiency, which confirmed the broad applicability of this system in . In summary, our method provides a reliable reverse-genetics platform that facilitates the study of stress-resistance mechanisms in , effectively overcoming previous technical limitations in genetic research on this species.
The online version contains supplementary material available at 10.1186/s13007-025-01427-z.
作为一种对非生物胁迫具有显著耐受性的盐生模式植物，拥有用于作物改良的宝贵遗传资源。然而，缺乏可靠且稳定的转化系统阻碍了该物种中候选基因的功能表征。在本研究中，我们通过构建基于烟草脆裂病毒（TRV）的靶向八氢番茄红素去饱和酶基因（）的载体，在中建立了一种高效的病毒诱导基因沉默（VIGS）系统，该基因的沉默会诱导出易于识别的光漂白表型。通过对中VIGS实施的接种材料和方法进行比较分析，我们发现通过真空辅助农杆菌浸润（0.5 kPa，10分钟）用携带TRV2：载体的悬浮液（OD = 0.8）接种发芽种子，进一步实现了约16.4%的平均沉默效率。接种后约15天，新出现的叶片出现系统性光漂白表型，qRT-PCR证实转录本丰度降低了40 - 80%。此外，我们通过沉默两个水通道蛋白基因（和）成功验证了该VIGS系统，实现了60.3 - 69.5%的敲低效率，这证实了该系统在中的广泛适用性。总之，我们的方法提供了一个可靠的反向遗传学平台，有助于研究中的抗逆机制，有效克服了该物种遗传研究中先前的技术限制。
在线版本包含可在10.1186/s13007-025-01427-z获取的补充材料。
