Shanghai Botanical Garden, Shanghai, China.
Sci Rep. 2020 Feb 12;10(1):2445. doi: 10.1038/s41598-020-58745-6.
There exist differences in the heat tolerance of Chinese rose varieties, and high temperature in summer can lead to failure of blooming in non-heat-tolerant Chinese rose varieties. We cloned a heat shock protein 70 gene (designated RcHSP70) from heat-tolerant varieties of Chinese rose (Rosa hybrida L.) to elucidate the molecular mechanism of heat tolerance and improve the quality of Chinese rose. Degenerate primers were designed for RcHSP70 according to the 5'- and 3'-end sequences of HSP70 genes in apple and tea. RcHSP70 was cloned from heat-tolerant Chinese rose varieties after heat shock. The heat shock-induced expression patterns of RcHSP70 in different Chinese rose varieties were analyzed by RT-PCR. Following heat shock (38 °C/3 h), RcHSP70 was highly expressed in the heat-tolerant varieties but not in the non-heat-tolerant varieties, indicating a close relationship between RcHSP70 and heat resistance in Chinese rose. To verify the function of RcHSP70, we constructed a prokaryotic expression recombinant vector for this gene and transformed it into Escherichia coli BL21. The tolerance of recombinant strains to abiotic stresses, including high temperature, low temperature, high salt, heavy metals, high pH, and oxidation, was evaluated. Additionally, RcHSP70 was transformed into tobacco plants. Because of the overexpression of this gene, transgenic tobacco plants improved their tolerance to high temperature and cold. In addition, transgenic tobacco showed better photosynthetic performance, relative electrical conductivity and proline content than wild tobacco after heat stress and cold stress. Our findings indicate that RcHSP70 is involved in the resistance of Chinese rose to abiotic stresses.
存在中国玫瑰品种耐热性的差异，夏季高温会导致不耐热的中国玫瑰品种开花失败。我们从耐热品种的中国玫瑰（Rosa hybrida L.）中克隆了一个热休克蛋白 70 基因（命名为 RcHSP70），以阐明耐热性的分子机制并提高中国玫瑰的品质。根据苹果和茶的 HSP70 基因的 5'和 3'端序列，为 RcHSP70 设计了简并引物。耐热中国玫瑰品种经热激后克隆 RcHSP70。通过 RT-PCR 分析不同中国玫瑰品种中 RcHSP70 的热激诱导表达模式。热激（38°C/3 h）后，耐热品种中 RcHSP70 高度表达，而非耐热品种中不表达，表明 RcHSP70 与中国玫瑰耐热性密切相关。为验证 RcHSP70 的功能，我们构建了该基因的原核表达重组载体并转化大肠杆菌 BL21。评估了重组菌株对非生物胁迫的耐受性，包括高温、低温、高盐、重金属、高 pH 值和氧化。此外，RcHSP70 被转化到烟草植物中。由于该基因的过表达，转基因烟草植物提高了对高温和低温的耐受性。此外，与野生型烟草相比，热激和冷激后转基因烟草表现出更好的光合性能、相对电导率和脯氨酸含量。我们的研究结果表明，RcHSP70 参与了中国玫瑰对非生物胁迫的抗性。
