Identification of Winter-Responsive Proteins in Bread Wheat Using Proteomics Analysis and Virus-Induced Gene Silencing (VIGS)

Abstract

Proteomic approaches were applied to identify protein spots involved in cold responses in wheat. By comparing the differentially accumulated proteins from two cultivars (UC1110 and PI 610750) and their derivatives, as well as the F 10 recombinant inbred line population differing in cold-tolerance, a total of 20 common protein spots representing 16 unique proteins were successfully identified using 2-DE method. Of these, 14 spots had significantly enhanced abundance in the cold-sensitive parental cultivar UC1110 and its 20 descendant lines when compared with the cold-tolerant parental cultivar PI 610750 and its 20 descendant lines. Six protein spots with reduced abundance were also detected. The identified protein spots are involved in stress/defense, carbohydrate metabolism, protein metabolism, nitrogen metabolism, energy metabolism, and photosynthesis. The 20 differentially expressed protein spots were chosen for quantitative real-time polymerase chain reaction (qRT-PCR) to investigate expression changes at the RNA level. The results indicated that the transcriptional expression patterns of 11 genes were consistent with their protein expression models. Among the three unknown proteins, Spot 20 (PAP6-like) showed high sequence similarities with PAP6. qRT-PCR results implied that cold and salt stresses increased the expression of PAP6-like in wheat leaves. Furthermore, VIGS (virus-induced gene silencing)-treated plants generated for PAP6-like were subjected to freezing stress, these plants had more serious droop and wilt, an increased rate of relative electrolyte leakage, reduced relative water content (RWC) and decreased tocopherol levels when compared with viral control plants. However, the plants that were silenced for the other two unknown proteins had no significant differences in comparison to the BSMV 0 -inoculated plants under freezing conditions. These results indicate that PAP6-like possibly plays an important role in conferring cold tolerance in wheat. Molecular & Cellular

Effects of T‐2 toxin on the muscle proteins of shrimp (Litopenaeus vannamei) – a proteomics study

“…The reflector mode was adopted, and peptides were selected in the mass range between 500 and 3500 Da with the resolution set to 50 000 dpi. The peptide mass finger printing (PMF) spectrum, which shows the m/z ( x ‐axis) data of different peptides was compared with the NCBI non‐redundant database of the Mascot search program (Matrix Science, London UK) to identify the proteins in the spots . The parameters are shown in Table .…”

“…In recent years, proteomics studies using techniques such as two‐dimensional electrophoresis (2‐DE) and mass spectrometry (MS) have become popular . Such studies provide an insight into the mechanisms associated with changes in muscle protein quality in shrimp . It has been suggested that proteomics could also be used to identify changes in muscle proteins of shrimp to better understand the pathogenesis of T‐2 …”

Temperature-induced changes in the wheat phosphoproteome reveal temperature-regulated interconversion of phosphoforms

“…So far, transcriptome and proteome profiles have been investigated in wheat under heat stress, revealing differences in gene expression and protein levels, respectively (Liu et al , 2015; Ahammed and Yu, 2016; Wang et al , 2016 b ; Zhang et al, 2016, 2017; Lu et al , 2017). However, protein post-translational modifications (PTMs) are linked with plant stresses, and reversible protein phosphorylation in particular functions as a crucial regulatory mechanism in many biological processes (Hashiguchi and Komatsu, 2016), including the regulation of abiotic stress signalling in crops (Kline et al , 2010; Bonhomme et al , 2012; Nguyen et al , 2012; Yang et al , 2013; Zhang et al , 2014a, b; Ahammed and Yu, 2016; Zhen et al , 2017).…”

Virus-induced gene silencing (VIGS) in cape gooseberry (Physalis peruviana L., Solanaceae)

“…However, these techniques have some disadavantages such as being relatively expensive, time-consuming and laborious [12]. Virus-induced gene silencing (VIGS) is a rapid tool for functional analyses used in different crops and non-model species such as wheat [13], tomato [14] and rye [15]. In addition, this method allows silencing different genes simultaneously in different plant organs like roots and leaves [16][17].…”