Cloning and Functional Analysis of the Soybean GmRIQ2 Promoter

Binbin Zhang,Qian Zhang,Xiaohui Song,Huayi Yin,Jing Deng,Zhihui Sun,Dongmei Li

doi:10.3390/agronomy12010227

Binbin Zhang, Qian Zhang + Show 5 more

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https://doi.org/10.3390/agronomy12010227

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Abstract

The RIQ gene can optimize the structure of light-harvesting complex II by regulating the degree of granum stacking, and ultimately participates in plant light stress responses. The GmRIQ2 gene contributes to plant photoprotection and may be involved in negative regulation of yield. To understand the regulatory mechanisms of GmRIQ2 expression, we isolated the GmRIQ2 promoter (PGmRIQ2) from the KenFeng 16 soybean genome. Sequence analysis showed that several cis-acting elements were involved in hormone responsiveness, seed-specific regulation, anaerobic induction and light responsiveness. PGmRIQ2 was expressed in soybean roots. GUS activity was detected in Arabidopsis thaliana rosette leaves, stems and roots, indicating that PGmRIQ2 tended to drive expression in rosette leaf vascular tissue and had spatiotemporal expression characteristics at the early growth stage. Three promoter deletion fragments of 450 bp, 284 bp and 194 bp were obtained by 5′ deletion. These three fragments all drove GUS gene expression in Arabidopsis thaliana rosette leaves, stems and roots. GUS gene expression decreased with deletion fragment truncation. In addition, the PGmRIQ2 auxin (IAA), abscisic acid (ABA) and methyl jasmonate (MeJA) response elements were located at base pairs −1661~−450, −450~−284 and −284~−194, respectively, and the promoter was induced by IAA, ABA, MeJA and light.

Highlights

IntroductionResearchers have focused on inducible promoters and tissue-specific promoters, hoping that they can replace constitutive promoters as regulators [3]
The results showed that there were four core promoter regions in the PGmRIQ2 sequence, which were located from base pairs −103 to −153, −516 to −566, −843 to −893, and −983 to −1033
Present were some cis-regulatory elements involved in abiotic stress responses, such as those involved in light responses; elements related to hormone responses (cis-acting elements involved in abscisic acid (ABA) responsiveness (ABA response elements, ABREs), a cis-acting regulatory element involved in IAA responsiveness, and cis-acting regulatory elements involved in methyl jasmonate (MeJA) responsiveness (CDTCA- and TGACG-motifs)); a cis-acting regulatory element essential for anaerobic induction; and a cis-acting regulatory element involved in seed-specific regulation

Summary

Introduction

Researchers have focused on inducible promoters and tissue-specific promoters, hoping that they can replace constitutive promoters as regulators [3] The former regulate the expression of foreign genes only in the presence of inducing conditions or factors and can be purposefully regulated according to desired outcomes [13,14]. The use of the latter can effectively alleviate unnecessary waste caused by overexpression because expression occurs only during a specific period or in a specific location [14,15,16]

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