Plant Cytogenomics Laboratory, University of Trás-os-Montes and Alto Douro (UTAD), Laboratorial Complex, Room A1.09, Quinta de Prados, 5000-801 Vila Real, Portugal.
Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro (UTAD), Laboratorial Complex, Quinta de Prados, 5000-801 Vila Real, Portugal.
Genes (Basel). 2024 Sep 30;15(10):1283. doi: 10.3390/genes15101283.
"Touriga Franca" (TF) and "Touriga Nacional" (TN) are grapevine varieties cultivated in the 'Douro Superior' subregion (Northern Portugal) that experience stressful environmental conditions during the summer.
Aiming to profile the expression of stress-responsive genes by quantitative real-time PCR (qPCR) in TF and TN plants growing naturally, three candidate reference genes were first tested under controlled conditions.
To simulate a summer's day, TF and TN in vitro plants were exposed to 32 °C-3 h (heat acclimation) and 42 °C-1 h (severe heat stress, HS) followed by two recovery periods (32 °C-3 h and 24 °C-24 h). Leaf samples were collected at the end of each phase. Control plants were kept at 24 °C.
Among the candidate reference genes, the and pair showed the highest stability. The suitability of these genes for qPCR was validated by () gene profiling. The expression was up-regulated in both varieties and all experimental phases except in TF control plants. TN showed the highest relative expression ratio after severe HS.
TN responded faster than TF to the induced heat shocks. The , , and genes revealed to be suitable for further qPCR assays in TF and TN grapevine varieties.
“Touriga Franca”（TF）和“Touriga Nacional”（TN）是在葡萄牙北部“Douro Superior”子产区种植的葡萄品种，它们在夏季会经历恶劣的环境条件。
旨在通过定量实时 PCR（qPCR）对自然生长的 TF 和 TN 植物中的应激响应基因表达进行分析，首先在受控条件下对三个候选参考基因进行了测试。
为了模拟夏日，将 TF 和 TN 体外植物暴露于 32°C-3 h（热驯化）和 42°C-1 h（严重热应激，HS），随后进行两个恢复期（32°C-3 h 和 24°C-24 h）。在每个阶段结束时采集叶片样本。对照植物保持在 24°C。
在候选参考基因中，和 对表现出最高的稳定性。通过（）基因分析验证了这些基因用于 qPCR 的适用性。在所有实验阶段，除了 TF 对照植物外，两种品种中的表达均上调。TN 在严重 HS 后表现出最高的相对表达比率。
TN 比 TF 对诱导的热冲击反应更快。在 TF 和 TN 葡萄品种中，、和基因被证明适用于进一步的 qPCR 分析。
