Glutathione S-transferase (GST) genes from transcripts of Vitis flexuosa leaves infected with Elsinoe ampelina were characterized and analyzed for their expression using primers based on specific regions. Comparison of deduced amino acid sequences from GST transcripts of V. flexuosa showed that the score of the deduced amino acid identity ranged from 43.38% (VfGST26625 and VfGST774) to 6.67% (Vf GST13892 and Vf GST774). Primary and secondary structure analysis was performed using the ProtParam and Self-Optimized Prediction Method with Alignment software. A phylogenetic tree was constructed from the GST proteins by the neighbor joining method using MEGA 6.0 to investigate the relationship among Vf GST, VvGST, and At GST proteins. To evaluate the differential expression pattern of GST genes by real-time polymerase chain reaction (PCR), primers specific to unique regions in each gene were obtained by alignment of the sequences. Real-time PCR revealed that GST genes were expressed differentially in the leaves of V. flexuosa infected with Botrytis cinerea, E. ampelina, and Rhizobium vitis. The expression of VfGST26625 was up-regulated, while that of others were down-regulated among five GSTs in all grapevine leaves inoculated with each pathogen. The results provided herein improve our understanding of defense responses to various pathogen attacks in grapevines.
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The differential expression of β-1,3-glucanase genes in the leaves of Vitis flexuosa induced by fungal and bacterial pathogen infections was investigated. The nucleotide and deduced amino acid sequences of β-1,3-glucanase genes from the transcripts of V. flexuosa were compared. The percentage similarity of deduced amino acid ranged from 22.0% between VfGlu34359 and VfGlu48103 to 96.9% in VvGlu2735 and VvGlu48103. To demonstrate the differential expression pattern of β-1,3-glucanase genes, primers specific to unique regions in each gene were obtained by alignment of the sequences. Accumulation patterns of β-1,3-glucanase mRNAs in the leaves of V. flexuosa were induced differentially and were dependent on the pathogens used including Botrytis cinerea, Colletotrichum acutatum, Elsinoe ampelina, and Rhizobium vitis. This study provides useful information that will improve our understanding of grapevine defense responses to various pathogen attacks.
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