Characterization of a strong and constitutive promoter from the Arabidopsis serine carboxypeptidase-like gene AtSCPL30 as a potential tool for crop transgenic breeding

Abstract

BackgroundTransgenic 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.ResultsIn 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.ConclusionsAs 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.Electronic supplementary materialThe online version of this article (10.1186/s12896-018-0470-x) contains supplementary material, which is available to authorized users.

Characterization of a novel LmSAP gene promoter from Lobularia maritima: Tissue specificity and environmental stress responsiveness

“…In consequence, promoters for multiple stress responses, such as PrLmSAP , could support the development of more stress-tolerant rice varieties. Second, plant response to stresses in a target environment must be fine-tuned; in the last decade remarkable progress has been made regarding the comprehension of stress-signalling networks in plants however the practical exploitation of this knowledge for transgene expression in plants is limited, partly due to the shortage of promoters with the desired expression (developmental- and tissue-specific or condition dependent) that allow for the fine-regulation of the target genes expression, especially in monocot species [ 16 , 42 , 76 ]. The results presented in this study position PrLmSAP as a good candidate for directing the expression of target tolerance-genes in rice in a tissue- and condition-specific manner.…”

Glutamine Synthetase (GS): A Key Enzyme for Nitrogen Assimilation in The Macroalga Gracilariopsis lemaneiformis (Rhodophyta)

“…Promoters play important roles in initiating gene transcription and regulating the expression of genes (Jiang et al. ). Thus, analysis of the transcriptional activity and expression pattern of a promoter is an important foundation for transgenic breeding and gene function studies (Hernandez‐Garcia et al.…”

“…The 5 0 end flanking regions of plant GS genes contain light-responsive elements, auxinresponsive elements, binding sites for transcription factors (TFs), and regulatory elements involved in tissue-specific expression (Castro-Rodr ıguez et al 2011a). Promoters play important roles in initiating gene transcription and regulating the expression of genes (Jiang et al 2018). Thus, analysis of the transcriptional activity and expression pattern of a promoter is an important foundation for transgenic breeding and gene function studies (Hernandez-Garcia et al 2009).…”

Biosensors: A Sneak Peek into Plant Cell’s Immunity

“…Expression under strong viral constitutive promoter CaMV 35S can experience silencing or patterns in dividing cells [ 7 ] and is therefore often exchanged with plant constitutive promoters, such as rice actin and maize ubiquitin promoters. Still, variable expression in different species forces the exploration of novel options [ 154 , 155 ]. On the other hand, it is possible to opt for separation of two or more reporters at the protein stage.…”