Cloning and Analysis of Stress Response Pattern of SlWRKY31 Gene Promoter from Tomato
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    Abstract:

    In this study, the wildtype tomato (Solanum lycopersicum) was used as the experimental material. The 1 849 bp promoter fragment in the upstream of the start codon of the SlWRKY31 gene from tomato was isolated by PCR cloning method. This promoter was then used to drive the GUS gene expression in wildtype tomato, and the obtained transgenic tomato was analysed by GUS histochemical staining and quantitative assay of GUS activity after different stress treatments. These results showed that: (1) sequence analysis revealed that the length of this promoter was 1 849 bp, and contained a large number of abiotic stess and phytohormone responsive elements, such as heat stess responsive element (HSE), droughtinducibility element (MBS), defense and stess responsive element (TCrich repeats), woundresponsive element (WUNmotif), abscisic acid responsive element (AREB), and salicylic acid responsive element (TGAelement). (2) Quantitative realtime PCR (qRTPCR) showed that the SlWRKY31 gene was expressed ubiquitously in all tissues examined, but its expression level was significantly increased under Mannitol, NaCl, SA, ABA, and heat stress (42 ℃) treatments. (3) A plant expression vector was constructed using the SlWRKY31 promoter to drive the GUS reporter gene (SlWRKY31Pro::GUS), and was transformed into wildtype tomato by Agrobacteriummediated transformation. GUS histochemical staining analysis of the transgenic tomato plants showed that the GUS gene was expressed in all tissues of tomato, indicating that the SlWRKY31 promoter is a constitutive promoter. (4) The GUS histochemical staining and the quantitative assay of transgenic plants after different stress treatments showed that the SlWRKY31 promoter was significantly induced by NaCl, Mannitol, SA, ABA and heat stress (42 ℃), indicating that the SlWRKY31 promoter is an inducible promoter that can respond to multiple stresses.

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GAO Yongfeng, YANG Fengming, LI Qinzhong, ZHANG Guoyan, WANG Dan, YAO Yinan, LIU Jikai. Cloning and Analysis of Stress Response Pattern of SlWRKY31 Gene Promoter from Tomato[J]. Acta Botanica Boreali-Occidentalia Sinica,2018,38(12):2155-2164

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  • Online: January 24,2019
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