Cloning and characterization of vacuolar Na+/H+ antiporter gene promoter from wheat (Triticum aestivum) cv. Kharchia Local

Soil salinity is one of the significant abiotic stresses in plants. Few efficient stress-inducible promoters from plants, especially from monocotyledonous crops like wheat (Triticum aestivum L.) are available to drive stress-inducible expression of transgene. Therefore, an experiment was conducted at ICAR-National Institute for Plant Biotechnology, New Delhi, during 2017–18 to clone and characterize stress-inducible promoter from wheat. Results found a 418 bp long NHX1 promoter (TaUNHX) from a bread wheat landrace, Kharchia Local. In silico analysis of TaUNHX predicted several cis-acting regulatory elements, including CAAT, DOF, GATA motifs and some essential stress-responsive elements. To analyze the activity of TaUNHX, Agrobacterium-mediated transient GUS assay in tobacco (Nicotiana tabacum L.) leaves and stems was studied under various abiotic stresses like salt (300 mM NaCl), drought (20% PEG) and ABA (100 μM). Stress-responsive nature of this promoter can be used to drive the expression of transgene following exposure to various stresses. The study's finding has significant implications as the characterized promoter can be used to develop transgenics where the transgene expression can be induced following exposure to stress.

[1]  Xue-Mei Kang,et al.  An efficient Agrobacterium-mediated transient transformation system and its application in gene function elucidation in Paeonia lactiflora Pall , 2022, Frontiers in Plant Science.

[2]  I. Muhammad,et al.  Cloning and functional analysis of the promoter of a stress-inducible gene (Zmap) in maize , 2019, PloS one.

[3]  Ke Zhang,et al.  Isolation and Functional Validation of Salinity and Osmotic Stress Inducible Promoter from the Maize Type-II H+-Pyrophosphatase Gene by Deletion Analysis in Transgenic Tobacco Plants , 2016, PloS one.

[4]  B. Jha,et al.  Functional Characterization of the Tau Class Glutathione-S-Transferases Gene (SbGSTU) Promoter of Salicornia brachiata under Salinity and Osmotic Stress , 2016, PloS one.

[5]  Dan Li,et al.  Molecular cloning of Phosphoethanolamine N-methyltransferase (PEAMT) gene and its promoter from the halophyte Suaeda liaotungensis and their response to salt stress , 2016, Acta Physiologiae Plantarum.

[6]  J. Verdeil,et al.  The promoter of the AlSAP gene from the halophyte grass Aeluropus littoralis directs a stress-inducible expression pattern in transgenic rice plants , 2015, Plant Cell Reports.

[7]  N. Tuteja,et al.  Cloning and functional characterization of the promoter of PsSEOF1 gene from Pisum sativum under different stress conditions using Agrobacterium-mediated transient assay , 2014, Plant signaling & behavior.

[8]  N. Tuteja,et al.  Isolation and functional characterization of the promoter of a DEAD-box helicase Psp68 using Agrobacterium-mediated transient assay , 2014, Plant signaling & behavior.

[9]  Yan Yan,et al.  GhWRKY40, a Multiple Stress-Responsive Cotton WRKY Gene, Plays an Important Role in the Wounding Response and Enhances Susceptibility to Ralstonia solanacearum Infection in Transgenic Nicotiana benthamiana , 2014, PloS one.

[10]  M. Fromm,et al.  Different gene-specific mechanisms determine the ‘revised-response’ memory transcription patterns of a subset of A. thaliana dehydration stress responding genes , 2014, Nucleic acids research.

[11]  E. Goyal,et al.  Isolation and functional characterization of Salt overly sensitive 1 (SOS1) gene promoter from Salicornia brachiata , 2013, Biologia Plantarum.

[12]  Chen Wang,et al.  Isolation and Characterization of an Endosperm-Specific Promoter from Wheat (Triticum aestivum L.) , 2012, Zeitschrift fur Naturforschung. C, Journal of biosciences.

[13]  K. Shinozaki,et al.  An ABRE promoter sequence is involved in osmotic stress-responsive expression of the DREB2A gene, which encodes a transcription factor regulating drought-inducible genes in Arabidopsis. , 2011, Plant & cell physiology.

[14]  S. Datta,et al.  Inducibility of three salinity/abscisic acid-regulated promoters in transgenic rice with gusA reporter gene , 2011, Plant Cell Reports.

[15]  Jukon Kim,et al.  Analysis of five novel putative constitutive gene promoters in transgenic rice plants , 2010, Journal of experimental botany.

[16]  M. Qi,et al.  In vivo analysis of plant promoters and transcription factors by agroinfiltration of tobacco leaves. , 2000, The Plant journal : for cell and molecular biology.

[17]  Q. Shen,et al.  Modular nature of abscisic acid (ABA) response complexes: composite promoter units that are necessary and sufficient for ABA induction of gene expression in barley. , 1996, The Plant cell.

[18]  Q. Shen,et al.  Functional dissection of an abscisic acid (ABA)-inducible gene reveals two independent ABA-responsive complexes each containing a G-box and a novel cis-acting element. , 1995, The Plant cell.

[19]  H. Pelham,et al.  Heat shock regulatory elements function as an inducible enhancer in the xenopus hsp70 gene and when linked to a heterologous promoter , 1986, Cell.

[20]  N. Li,et al.  Isolation and characterization of a tonoplast Na+/H+ antiporter from the halophyte Nitraria sibirica , 2015, Biologia Plantarum.

[21]  B. Jha,et al.  The transcriptional regulatory mechanism of the peroxisomal ascorbate peroxidase (pAPX) gene cloned from an extreme halophyte, Salicornia brachiata. , 2014, Plant & cell physiology.

[22]  Yoshiyuki Tanaka,et al.  Molecular and functional analyses of rice NHX-type Na+/H+ antiporter genes , 2010, Planta.