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Cloning and functional assessment of the protein disulfide isomerase (PDI-V) gene and its promoter sequences from Haynaldia villosa

Protein disulfide isomerase (PDI) and PDI-like genes encode key foldase enzymes that play an important role in disulfide bond formation, isomerization, and many other metabolic functions in plants. Our previous results indicate that the PDI-V expression is tissue-specific and that it is upregulated by powdery mildew and other abiotic stress treatments. | Turkish Journal of Botany Turk J Bot (2018) 42: 159-171 © TÜBİTAK doi:10.3906/bot-1701-34 http://journals.tubitak.gov.tr/botany/ Research Article Cloning and functional assessment of the protein disulfide isomerase (PDI-V) gene and its promoter sequences from Haynaldia villosa 1 1 1 1 2 1, Muhammad FAHEEM , Muhammad ARSHAD , Jin XIAO , Haiyan WANG , Yingbo LI , Xiue WANG * 1 The State Key Laboratory of Crop Genetics and Germplasm Enhancement, Cytogenetics Institute, Nanjing Agricultural University/JCIC-MCP, Nanjing, Jiangsu, P.R. China 2 Biotech Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai Key Laboratory of Agricultural Genetics and Breeding, Shanghai, P.R. China Received: 19.01.2017 Accepted/Published Online: 27.09.2017 Final Version: 20.03.2018 Abstract: Protein disulfide isomerase (PDI) and PDI-like genes encode key foldase enzymes that play an important role in disulfide bond formation, isomerization, and many other metabolic functions in plants. Our previous results indicate that the PDI-V expression is tissue-specific and that it is upregulated by powdery mildew and other abiotic stress treatments. In the current study, the complete PDI-V gene and its promoter sequence were cloned to identify gene structure and the important cis-regulating motifs responsible for various genes’ activities. Sequence analysis revealed the size of the cloned genomic sequence and promoter region as 5728 bp and 2086 bp, respectively. The PDI-V gene had nine exons and eight introns; this showed a highly conserved gene structure with that of wheat TaPDIL-5 in terms of exon size and number of exons/introns. The phylogenetic analysis of PDI-V with orthologue sequences from the A, B, and D genome of wheat, barley, and rice clearly demonstrated that the gene was highly conserved across the grass family. In silico analysis of the PDI-V promoter revealed a large number of elements responsible for endosperm or embryo specific expression, and .