Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Isolation and Characterization of Erianthus arundinaceus Phosphate Transporter 1 (PHT1) Gene Promoter and 5′ Deletion Analysis of Transcriptional Regulation Regions under Phosphate Stress in Transgenic Tobacco

Version 1 : Received: 3 October 2023 / Approved: 5 October 2023 / Online: 5 October 2023 (12:04:13 CEST)

A peer-reviewed article of this Preprint also exists.

Naveenarani, M.; Swamy, H.K.M.; Surya Krishna, S.; Mahadevaiah, C.; Valarmathi, R.; Manickavasagam, M.; Arun, M.; Hemaprabha, G.; Appunu, C. Isolation and Characterization of Erianthus arundinaceus Phosphate Transporter 1 (PHT1) Gene Promoter and 5′ Deletion Analysis of Transcriptional Regulation Regions under Phosphate Stress in Transgenic Tobacco. Plants 2023, 12, 3760. Naveenarani, M.; Swamy, H.K.M.; Surya Krishna, S.; Mahadevaiah, C.; Valarmathi, R.; Manickavasagam, M.; Arun, M.; Hemaprabha, G.; Appunu, C. Isolation and Characterization of Erianthus arundinaceus Phosphate Transporter 1 (PHT1) Gene Promoter and 5′ Deletion Analysis of Transcriptional Regulation Regions under Phosphate Stress in Transgenic Tobacco. Plants 2023, 12, 3760.

Abstract

Phosphorus deficiency highly interferes with plant growth and development. Plants respond to the persistent P deficiency by coordinating the expression of genes involved in the alleviation of the stress. Promoters of phosphate transporter genes are of great choice for the development of genetically modified plants with enhanced phosphate uptake abilities, which improve crop yields in phosphate-deficient soils. In our previous study, the sugarcane phosphate transporter PHT1;2 gene, showed a significantly high expression under salinity stress. In this study, the Erianthus arundinaceus EaPHT1;2 gene was isolated and characterized using various in-silico tools. The deduced 542 amino acid residues have 10 transmembrane domains, with a molecular weight and isoelectric point of 58.9 kDa and 9.80, respectively. They displayed 71–96% similarity with Arabidopsis thaliana, Zea mays, and Saccharum hybrid. To elucidate the function of the 5’ regulatory region, the 1.1kb promoter was isolated and validated in tobacco transgenics under Pi stress. The EaPHT1;2 promoter activity was detected using the β-glucuronidase (GUS) assay. While the EaPHT1;2 promoter showed 3 to 4.2-fold higher expression than the most widely used CaMV35S promoter. The 5’ deletion analysis with and without 5’UTR regions, revealed that a small size 374 bp fragment with the highest promoter activity among 5′ truncated fragments, which was 2.7 and 4.2 times higher than the well-used CaMV35S promoter under normal and Pi-deprivation conditions, respectively. The strong and short promoter of EaPHT1;2 with 374 bp showed significant expression in low Pi-stress conditions and it could be a valuable source for the development of stress-tolerant transgenic crops

Keywords

Erianthus arundinaceus; High-affinity phosphate transporter; PHT1; 2; Promoter analysis; Tobacco transgenic; Pi stress

Subject

Environmental and Earth Sciences, Soil Science

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