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Identification of upstream transcription factors and an interacting PP2C protein of dehydrin WZY2 gene in wheat.

ABSTRACT: As functional proteins dehydrins are found in many maturing seeds and vegetable tissue under adverse environmental conditions. However, the regulation of dehydrin expression remains unclear.To explore regulatory mechanisms of wheat dehydrin WZY2 expression under abiotic stresses, we constructed a cDNA library from PEG- and cold-treated wheat seedlings and performed yeast one-hybrid assay and yeast two-hybrid assay to identify the upstream transcription factors and protein interacting with dehydrin WZY2 gene. Yeast one-hybrid assay illustrated that bHLH49-like (GenBank NO. XM_020296294), zinc finger A20 and AN1 domain-containing stress-associated protein 6-like (GenBank NO. XM_020341647), and bHLH47-like (GenBank NO. XM_020313116) proteins can bind and interact with the promoter of the WZY2, these sequences are Aegilops tauschii transcription factors, the diploid progenitor of the D genome of hexaploid wheat (Triticum aestivum, genomes AABBDD). Real-time PCR analyzes unraveled the stress responsive expression of XM_020296294, XM_020341647, and XM_020313116 in wheat. XM_020296294 and XM_020341647 showed a similar expression patterns with WZY2. Yeast two-hybrid assay indicated that PP2C (GenBank NO. XM_020293398) protein can interact with WZY2. These results provided evidences that WZY2 could be positively regulated by XM_020296294 and XM_020341647 transcription factors, and WZY2 may also play an important role in the ABA signaling pathway through interaction with PP2C to regulate stress-responsive genes expression in wheat. The obtained results contribute for provide a better understanding of the regulatory mechanism of dehydrin expression under abiotic stresses in wheat.

SUBMITTER: Liu H

PROVIDER: S-EPMC6866685 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Identification of upstream transcription factors and an interacting PP2C protein of dehydrin <i>WZY2</i> gene in wheat.

Liu Hao H Yang Ying Y Zhang Linsheng L

Plant signaling & behavior 20191012 12

As functional proteins dehydrins are found in many maturing seeds and vegetable tissue under adverse environmental conditions. However, the regulation of dehydrin expression remains unclear.To explore regulatory mechanisms of wheat dehydrin WZY2 expression under abiotic stresses, we constructed a cDNA library from PEG- and cold-treated wheat seedlings and performed yeast one-hybrid assay and yeast two-hybrid assay to identify the upstream transcription factors and protein interacting with dehydr ...[more]

PMID: 31608778

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Project description:RWP-RKs represent a small family of transcription factors (TFs) that are unique to plants and function particularly under conditions of nitrogen starvation. These RWP-RKs have been classified in two sub-families, NLPs (NIN-like proteins) and RKDs (RWP-RK domain proteins). NLPs regulate tissue-specific expression of genes involved in nitrogen use efficiency (NUE) and RKDs regulate expression of genes involved in gametogenesis/embryogenesis. During the present study, using in silico approach, 37 wheat RWP-RK genes were identified, which included 18 TaNLPs (2865 to 7340 bp with 4/5 exons), distributed on 15 chromosomes from 5 homoeologous groups (with two genes each on 4B,4D and 5A) and 19 TaRKDs (1064 to 5768 bp with 1 to 6 exons) distributed on 12 chromosomes from 4 homoeologous groups (except groups 1, 4 and 5); 2-3 splice variants were also available in 9 of the 37 genes. Sixteen (16) of these genes also carried 24 SSRs (simple sequence repeats), while 11 genes had targets for 13 different miRNAs. At the protein level, MD simulation analysis suggested their interaction with nitrate-ions. Significant differences were observed in the expression of only two (TaNLP1 and TaNLP2) of the nine representative genes that were used for in silico expression analysis under varying levels of N at post-anthesis stage (data for other genes was not available for in silico expression analysis). Differences in expression were also observed during qRT-PCR, when expression of four representative genes (TaNLP2, TaNLP7, TaRKD6 and TaRKD9) was examined in roots and shoots of seedlings (under different conditions of N supply) in two contrasting genotypes which differed in NUE (C306 with low NUE and HUW468 with high NUE). These four genes for qRT-PCR were selected on the basis of previous literature, level of homology and the level of expression (in silico study). In particular, the TaNLP7 gene showed significant up-regulation in the roots and shoots of HUW468 (with higher NUE) during N-starvation; this gene has already been characterized in Arabidopsis and tobacco, and is known to be involved in nitrate-signal transduction pathway.

Dataset Information

Identification of upstream transcription factors and an interacting PP2C protein of dehydrin WZY2 gene in wheat.

Publications

Identification of upstream transcription factors and an interacting PP2C protein of dehydrin <i>WZY2</i> gene in wheat.

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