Quantitative analysis of proteome extracted from barley crowns grown under different drought conditions

Abstract

Barley cultivar Amulet was used to study the quantitative proteome changes through different drought conditions utilizing two-dimensional difference gel electrophoresis (2D-DIGE). Plants were cultivated for 10 days under different drought conditions. To obtain control and differentially drought-treated plants, the soil water content was kept at 65, 35, and 30% of soil water capacity (SWC), respectively. Osmotic potential, water saturation deficit, 13C discrimination, and dehydrin accumulation were monitored during sampling of the crowns for proteome analysis. Analysis of the 2D-DIGE gels revealed 105 differentially abundant spots; most were differentially abundant between the controls and drought-treated plants, and 25 spots displayed changes between both drought conditions. Seventy-six protein spots were successfully identified by tandem mass spectrometry. The most frequent functional categories of the identified proteins can be put into the groups of: stress-associated proteins, amino acid metabolism, carbohydrate metabolism, as well as DNA and RNA regulation and processing. Their possible role in the response of barley to drought stress is discussed. Our study has shown that under drought conditions barley cv. Amulet decreased its growth and developmental rates, displayed a shift from aerobic to anaerobic metabolism, and exhibited increased levels of several protective proteins. Comparison of the two drought treatments revealed plant acclimation to milder drought (35% SWC); but plant damage under more severe drought treatment (30% SWC). The results obtained revealed that cv. Amulet is sensitive to drought stress. Additionally, four spots revealing a continuous and significant increase with decreasing SWC (UDP-glucose 6-dehydrogenase, glutathione peroxidase, and two non-identified) could be good candidates for testing of their protein phenotyping capacity together with proteins that were significantly distinguished in both drought treatments.

Barley (Hordeum vulgare L.) Improvement Past, Present and Future

“…The analysis indicated that the differentially regulated proteins were related to photosynthesis, detoxification, energy metabolism and protein biosynthesis. Barley, also, used to identify the quantitative proteome changes under different drought conditions by Vítámvás et al [184]. They cultivated plants for 10 days under different drought conditions that the soil water content was held at 65, 35 and 30% of soil water capacity (SWC), respectively.…”

Physiology and proteomics of two maize genotypes with different drought resistance

“…Furthermore, at the molecular level, drought stress affects gene expression level of different pathways related to stress perception, signal transduction, regulators, and the synthesis of stress-related proteins (Kakumanu et al 2012). Proteomic changes of many plants have been studied under drought stress, including wheat (Faghani et al 2015), barley (Vítámvás et al 2015), cotton (Deeba et al 2012), tobacco (Xie et al 2016), rapeseeds (Urban et al 2016), peanut (Katam et al 2016), grapevine (Król and Weidner 2017), and maize (Benesova et al 2012, Hu et al 2011, Liu et al 2012. Studies of proteins in maize (Kakumanu et al 2012) and wheat (Caruso et al 2009) show that a very close relationship exists between drought resistance and gene expression.…”

Optimized Western Blotting Exploration of H3K27 Methylation Changes under Waterlogging in Barley

“…Likewise, the quality of the total protein extracted from the same plant organ using different methods may differ to a certain extent (Saravanan and Rose, 2004;Abdullah et al, 2017). Although research studies have extracted protein from barley, these studies have mainly relied on twodimensional gel electrophoresis (Vítámvás et al, 2015;Guo et al, 2016). Moreover, these reported methods are costly and time-consuming (Ghosh et al, 2014;Wu et al, 2014), easy and stable protein extraction methods for western blotting remain unexplored in barley.…”