Abstract

Salinity and drought are two major abiotic stresses that limit grape productivity. Responses to stress in grape are known to be regulated by several families of transcription factors. However, little is known about the role of grape Squamosa promoter binding protein (SBP)-box transcription factor genes in response to abiotic stress. To better understand the functions of the grape SBP-box genes in abiotic stress tolerance, a full-length complementary DNA (cDNA) sequence of the putative SBP-box transcription factor gene, VpSBP16 was amplified from Chinese wild grapevine Vitis pseudoreticulata clone “Baihe-35-1”. We observed that the VpSBP16 protein fused to the green fluorescent protein (GFP) reporter accumulated in the nucleus when transiently expressed in onion epidermal cells. Moreover, VpSBP16 was shown to have transcriptional activation activity using a yeast trans-activation assay. We performed a VpSBP16 functional analysis through the characterization of transgenic Arabidopsis thaliana plants constitutively over-expressing VpSBP16. The transgenic lines had longer roots and the seeds had a higher germination rate than the wild type (WT) under osmotic stress. In addition, the accumulation of reactive oxygen species (ROS) of transgenic seedlings was significantly lower than WT in the transgenic lines, as was electrolyte leakage. VpSBP16 overexpression also elevated expression levels of stress-response genes involved in the salt overly sensitive (SOS) pathway. These results indicate that overexpression VpSBP16 in A. thaliana enhances tolerance of salt and drought stress during seed germination, as well in seedlings and mature plants, by regulating SOS and ROS signaling cascades.

Highlights

  • Plants frequently encounter stressful environmental conditions, such as extreme temperatures, drought, and high salinity, which can severely limit growth and development and greatly reduce the quality and yield of crops

  • We describe the functional evaluation of an Squamosa promoter binding protein (SBP) gene, VpSBP16, from a Vitis wild species following its over expression in A. thaliana, and present data that support a role of its role enhancing abiotic stress resistance

  • Wang clone ‘Baihe-35-1’, using the primers based on the complementary DNA (cDNA) sequence of VvSBP16 obtained from the Grape Genome Database

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Summary

Introduction

Plants frequently encounter stressful environmental conditions, such as extreme temperatures, drought, and high salinity, which can severely limit growth and development and greatly reduce the quality and yield of crops. Transcriptional control of the expression of stress-responsive genes is a crucial part of plant responses to abiotic stress and many transcription factors have been identified that regulate responses to environmental cues by activating or repressing multiple target genes. MYB2, MYB15 [1,2]; bZIP24 [3]; WRKY25, WRKY33, WRKY63/ABO3, WRKY18, WRKY60 [4,5,6] from Arabidopsis thaliana, OsMPS [7] from rice (Oryza sativa) and SlAREB [8] from tomato (Solanum lycopersicum) have all been reported to regulate responses to some abiotic stresses by activating or repressing transcription of multiple target genes, such as RD29A, COR15, KIN1. We describe the functional evaluation of an SBP gene, VpSBP16, from a Vitis wild species following its over expression in A. thaliana, and present data that support a role of its role enhancing abiotic stress resistance

Cloning and Sequence Analysis of VpSBP16
F: AFG:GACGAGCCCATCTCTTACATCTACACTCACACATAGCAAGC
F: CCF:ACTCGAGTTGAGATTAGAATAGGACATAGCCTTGATCCTTGTTCCTTATTCTAT R
Plant Materials and Growth Conditions
Isolation and Analysis of the VpSBP16 cDNA
Subcellular Localization and Trans-Activation Assay
Semi-Quantitative RT-PCR Analysis
Germination Assays
Determination of the Water Loss Rate and Electrolyte Leakage
4.10. Quantitative Real-Time RT-PCR Analysis
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