101. Characterization of a strong and constitutive promoter from the Arabidopsis serine carboxypeptidase-like gene AtSCPL30 as a potential tool for crop transgenic breeding
- Author
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Kewei Zhang, Kunpeng Li, Qiuxia He, Ke Zhang, Wendi Li, Jiang Pingping, Wen Cheng, Zhaohua Ding, and Shuangfeng Zhu
- Subjects
0106 biological sciences ,0301 basic medicine ,lcsh:Biotechnology ,Transgene ,Stress treatment ,Molecular Sequence Data ,Arabidopsis ,Gene Expression ,Nicotiana benthamiana ,Carboxypeptidases ,Genetically modified crops ,01 natural sciences ,03 medical and health sciences ,Plasmid ,GUS analysis ,lcsh:TP248.13-248.65 ,Tobacco ,Constitutive promoter ,Promoter Regions, Genetic ,Enhancer ,Gene ,Base Sequence ,biology ,Arabidopsis Proteins ,fungi ,food and beverages ,Promoter ,Plants, Genetically Modified ,biology.organism_classification ,AtSCPL30 ,Cell biology ,Transformation (genetics) ,030104 developmental biology ,CaMV35S ,010606 plant biology & botany ,Biotechnology - Abstract
Background Transgenic technology has become an important technique for crop genetic improvement. The application of well-characterized promoters is essential for developing a vector system for efficient genetic transformation. Therefore, isolation and functional validation of more alternative constitutive promoters to the CaMV35S promoter is highly desirable. Results In this study, a 2093-bp sequence upstream of the translation initiation codon ATG of AtSCPL30 was isolated as the full-length promoter (PD1). To characterize the AtSCPL30 promoter (PD1) and eight 5′ deleted fragments (PD2-PD9) of different lengths were fused with GUS to produce the promoter::GUS plasmids and were translocated into Nicotiana benthamiana. PD1-PD9 could confer strong and constitutive expression of transgenes in almost all tissues and development stages in Nicotiana benthamiana transgenic plants. Additionally, PD2-PD7 drove transgene expression consistently over twofold higher than the well-used CaMV35S promoter under normal and stress conditions. Among them, PD7 was only 456 bp in length, and its transcriptional activity was comparable to that of PD2-PD6. Moreover, GUS transient assay in the leaves of Nicotiana benthamiana revealed that the 162-bp (− 456~ − 295 bp) and 111-bp (− 294~ − 184 bp) fragments from the AtSCPL30 promoter could increase the transcriptional activity of mini35S up to 16- and 18-fold, respectively. Conclusions As a small constitutive strong promoter of plant origin, PD7 has the advantage of biosafety and reduces the probability of transgene silencing compared to the virus-derived CaMV35S promoter. PD7 would also be an alternative constitutive promoter to the CaMV35S promoter when multigene transformation was performed in the same vector, thereby avoiding the overuse of the CaMV35S promoter and allowing for the successful application of transgenic technology. And, the 162- and 111-bp fragments will also be very useful for synthetic promoter design based on their high enhancer activities.
- Published
- 2018