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| Functional analysis of stressrelated transcription factor gene PsDREB from Paeonia suffruticosa |
| LIU Huichun*, MA Guangying, ZHU Kaiyuan, ZOU Qingcheng, ZHOU Jianghua, TIAN Danqing |
| (Flower Research and Development Center, Zhejiang Academy of Agricultural Sciences, Hangzhou 311202,China) |
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Abstract Peony (Paeonia suffruticosa) is one of the most famous traditional flowers with profound cultural connotation in China. With the fast development of gardenflower industry, the application of peony increased dramatically for the purpose of landscaping or medicinal uses. However, stress resistance of peony has been poor due to artificial domestication for thousands of years, which greatly restricted the development of peony. With the significant interest of adding new vitality into the flower industry by successfully introducing and cultivating peony in southern China, improving the tolerance of peony cultivars to stress has become crucial. The dehydration responsive element binding protein (DREB) gene is one of the most important transcription factors that was found to be related to adversity resistance. The objective of this study was to analyze the biological function of DREBgene from peony, named as PsDREB, with methods of yeast onehybrid assay and transgenic technology.
In this study, the ability of PsDREB protein binding with DRE and its transcriptional activation activity were tested by yeast onehybrid assay. Results showed that the PsDREB protein was able to combine specifically with the DRE, but unable to combine with the mutated DRE (mDRE), showing a strong transcriptional activity. Subsequently, the PsDREB gene was transferred into tobacco and was mediated by Agrobacterium to determine its biological function in response to adversity. Thereafter, the tolerance ability of the transgenic tobacco was detected under manipulated drought, low temperature, high salt, and abscisic acid (ABA) treatments, as compared to controls.
In conclusion, the PsDREB protein has strong binding and transcriptional activities. The overexpression of PsDREBcan obviously enhance plant resistance to abiotic stresses, especially for drought and high salt treatments. These results lay the foundation for further research on the function mode of PsDREB transcription factor, and also supply theoretical and material evidence for excavating more possible stressresistant genes from peony so as to breed out highly stresstolerant transgenic peony.
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Received: 15 May 2015
Published: 31 August 2016
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牡丹抗逆转录因子基因PsDREB的功能解析
通过酵母单杂交试验，对牡丹抗逆转录因子基因PsDREB的DNA结合能力及转录激活功能进行分析。结果显示：PsDREB蛋白能够特异性地与顺式作用元件DRE（dehydration responsive element，干旱应答元件）结合，但不能与突变的DRE(mDRE)结合；该蛋白能够激活下游报告基因His的表达，从而使酵母细胞在缺失His的培养基上能够正常生长。说明PsDREB具有转录激活功能。为了进一步研究该转录因子基因PsDREB在植物抗逆反应中的功能，利用农杆菌介导法将其转化到模式植物烟草中，对获得的转基因烟草进行低温、干旱、高盐、脱落酸等胁迫处理。结果发现：与对照相比，超量表达PsDREB基因能够明显提高烟草对逆境胁迫的抗性，尤其是在抗干旱和高盐方面效果显著。
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