Characterization of Erysiphe necator-Responsive Genes in Chinese Wild Vitis quinquangularis

Abstract

Powdery mildew (PM), caused by fungus Erysiphe necator, is one of the most devastating diseases of grapevine. To better understand grapevine-PM interaction and provide candidate resources for grapevine breeding, a suppression subtractive hybridization (SSH) cDNA library was constructed from E. necator-infected leaves of a resistant Chinese wild Vitis quinquangularis clone “Shang-24”. A total of 492 high quality expressed sequence tags (ESTs) were obtained and assembled into 266 unigenes. Gene ontology (GO) analysis indicated that 188 unigenes could be assigned with at least one GO term in the biological process category, and 176 in the molecular function category. Sequence analysis showed that a large number of these genes were homologous to those involved in defense responses. Genes involved in metabolism, photosynthesis, transport and signal transduction were also enriched in the library. Expression analysis of 13 selected genes by qRT-PCR revealed that most were induced more quickly and intensely in the resistant material “Shang-24” than in the sensitive V. pseudoreticulata clone “Hunan-1” by E. necator infection. The ESTs reported here provide new clues to understand the disease-resistance mechanism in Chinese wild grapevine species and may enable us to investigate E. necator-responsive genes involved in PM resistance in grapevine germplasm.

Differential expression of molecular rust resistance components have distinctive profiles in Coffea arabica - Hemileia vastatrix interactions

“…Chalcone synthase and polyubiquitin were the other genes upregulated during incompatible interaction with defense or cell maintenance functions (Fernandez et al 2004). No ABC types of resistance protein transporter genes were induced in both interactions despite these proteins were reported in different coffee cultivars (Guzzo et al 2009) and Vitis quinquangularis against Erysiphe necator (M. Gao et al 2012).…”

“…which could route to activation of other signaling pathways as seen in A. thaliana (Kovtun et al 2000). Different types of kinases and GTPbinding proteins known to characterize upregulated libraries in both interactions as seen in different cultivars and other plants (Guzzo et al 2009;Medeiros et al 2009;Gao et al 2012). A large number of genes involved in signal transduction were downregulated at 12 and 24 h.a.i.…”

Overexpression of a SBP-Box Gene (VpSBP16) from Chinese Wild Vitis Species in Arabidopsis Improves Salinity and Drought Stress Tolerance

“…Quantitative RT-PCR was conducted using SYBR green (Takara Biotechnology) with an IQ5 real time PCR machine (Bio-Rad, Hercules, CA, USA). Each reaction was performed in triplicate and data were analyzed as previously described [ 54 ]. The expression of stress-related genes such as AtFRY1, AtSAD1, AtADH, AtP5CS1, AtRD29B, AtCDPK1 and AtCDPK2 were measured by RT-PCR in WT and T3 transgenic lines.…”

The SBP-Box Gene VpSBP11 from Chinese Wild Vitis Is Involved in Floral Transition and Affects Leaf Development

“…The 25 µL PCR reaction contained 12.5 µL of SYBR ® Premix Ex Taq TM II (2×), 1 µL of PCR forward primer (10 µm), 1 µL of PCR reverse primer (10 µm), 2 µL of 10× diluted cDNA, and 8.5 µL of ddH 2 O. Cycling parameters were 95 °C for 30 s, 40 cycles of 95 °C for 5 s, and 60 °C for 30 s. For dissociation curve analysis, a program including 95 °C for 15 s, followed by a constant increase from 60 to 95 °C was included after the PCR cycles. Each reaction was done in triplicate and was analyzed using the protocol described by Gao et al [ 42 ].…”