Promoter activity analysis and transcriptional profile of Ginkgo biloba 1-Deoxy-D- Xylulose 5-Phosphate reductoisomerase gene (GbDXR) under abiotic stresses

Abstract

Terpene trilactones (TTL) is a pharmacological ingredient in Ginkgo biloba and its content has become one of the key indices for medicinal value evaluation of ginkgo. 1-Deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) catalyzes the first step specific for isopentenyl diphosphate production in methylerythritol phosphate pathway, which provide the basic structure required for TTLs biosynthesis. To understand the mechanism controlling the GbDXR gene expression, the GbDXR promoter sequence was isolated and subjected to transient expression with the green fluorescent protein (GFP) in tobacco plants. Characteristic analysis revealed various cis-acting elements that related to light-regulated transcription, hormone signaling (auxin, ethylene), adversity stress and defense signaling (heat/dehydration stress) in the GbDXR promoter region. In transient expression assay, deletion of different portions of the upstream GbDXR promoter identified that the promoter region -3230bp to -865bp conserve the positive regulation function, which could promote the expression of GFP in the cytoplasm of tobacco leaf epidermal cells. The regulation function of the promoter region -865bp to -262bp remained to be elucidated. EMSA analysis suggested possible interactions of GbERF10 and GbERF17 with the ERF-binding elements in the upstream of GbDXR promoter. For abiotic stresses treatment, the expression of GbDXR gene could be significantly induced by UV-B and drought stress. In general, the GbDXR gene expressed differently in different ginkgo tissues but exhibited the highest transcriptional level in the root, with the maximum TTLs content simultaneously. The positive relationship between gene expression level and TTLs content indicated that the GbDXR is responsible for TTLs biosynthesis in G. biloba.

Treatment of Ginkgo biloba with Exogenous Sodium Selenite Affects Its Physiological Growth, Changes Its Phytohormones, and Synthesizes Its Terpene Lactones

“…DXR catalyzes DXP to generate MEP, which is an important rate limiting enzyme in the MEP pathway and an ideal target for regulating the MEP pathway [ 8 ]. In a study focusing on the DXR gene promoter of G. biloba , we found that WRKY, MYB, MYC, and AP2/ERF binding sites exist in its upstream regulatory elements, and AP2/ERF are important TFs regulating the expression of the DXR gene [ 39 ]. WRKYs were found to be involved in the regulation of other terpenoid synthesis structure genes in G. biloba .…”