Chen Fangyuan, Wang Lai, Huang Qixiu, Jiang Run, Li Wenhui, Hou Xianfei, Tan Zihan, Lei Zhonghua, Li Qiang, Zeng Youling
Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830017, China.
Crop Research Institute, Xinjiang Uygur Autonomous Region Academy of Agricultural Sciences, Urumqi 830091, China.
Plants (Basel). 2025 Aug 4;14(15):2412. doi: 10.3390/plants14152412.
Sunflower ( L.) is an important oilseed crop in Northwest China, exhibiting resistance to salt and drought. Mining its excellent tolerance genes can be used for breeding. However, the current platforms for identifying gene function in sunflower is inadequate. The transient transformation system, which can rapidly validate gene function, shows promising prospects in research. In this study, we established an efficient transient expression transformation system for sunflower using three methods: -mediated infiltration, injection, and ultrasonic-vacuum. The detailed procedures were as follows: GV3101 carrying a reporter gene on the pBI121 vector with an OD of 0.8 as the bacterial suspension and 0.02% Silwet L-77 as the surfactant were utilized in all three approaches. For the infiltration method, seedlings grown hydroponically for 3 days were immersed in a bacterial suspension containing 0.02% Silwet L-77 for 2 h; for the injection method, the same solution was injected into the cotyledons of seedlings grown in soil for 4 to 6 days. Subsequently, the seedlings were cultured in the dark at room temperature for three days; for the ultrasonic-vacuum method, seedlings cultured in Petri dishes for 3 days were first subjected to ultrasonication at 40 kHz for 1 min, followed by vacuum infiltration at 0.05 kPa for 5-10 min. -mediated transient transformation efficiency achieved by the three methods exceeded 90%, with gene expression being sustained for at least 6 days. Next, we employed the infiltration-based sunflower transient transformation technology with the stable transformation platform to confirm salt and drought stress tolerance of candidate gene from sunflower responding to various abiotic stresses. Altogether, this study successfully established an -mediated transient transformation system for sunflower using these three methods, which can rapidly identify gene function and explore the molecular mechanisms underlying sunflower's resistance traits.
向日葵（L.）是中国西北地区一种重要的油料作物，具有耐盐和耐旱性。挖掘其优良的耐受基因可用于育种。然而，目前用于鉴定向日葵基因功能的平台并不完善。能够快速验证基因功能的瞬时转化系统在研究中显示出广阔的前景。在本研究中，我们采用三种方法建立了一种高效的向日葵瞬时表达转化系统：农杆菌介导的浸润法、注射法和超声-真空法。具体步骤如下：在所有三种方法中，均使用携带pBI121载体上报告基因的GV3101，其OD值为0.8作为细菌悬浮液，并使用0.02%的Silwet L-77作为表面活性剂。对于浸润法，将水培3天的幼苗浸入含有0.02% Silwet L-77的细菌悬浮液中2小时；对于注射法，将相同溶液注射到土培4至6天的幼苗子叶中。随后，将幼苗在室温黑暗条件下培养3天；对于超声-真空法，将在培养皿中培养3天的幼苗先在40 kHz下超声处理1分钟，然后在0.05 kPa下真空浸润5至10分钟。三种方法实现的农杆菌介导的瞬时转化效率均超过90%，基因表达持续至少6天。接下来，我们将基于浸润的向日葵瞬时转化技术与稳定转化平台相结合，以确认向日葵候选基因对各种非生物胁迫的盐和干旱胁迫耐受性。总之，本研究成功地利用这三种方法建立了一种向日葵农杆菌介导的瞬时转化系统，该系统可以快速鉴定基因功能并探索向日葵抗性性状的分子机制。
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