Here, we describe a method for the combined metabolomic, proteomic, transcriptomic and genomic analysis from one single sample as a major step for multilevel data integration strategies in systems biology. While extracting proteins and DNA, this protocol also allows the separation of metabolites into polar and lipid fractions, as well as RNA fractionation into long and small RNAs, thus allowing a broad range of transcriptional studies. The isolated biomolecules are suitable for analysis with different methods that range from electrophoresis and blotting to state-of-the-art procedures based on mass spectrometry (accurate metabolite profiling, shot-gun proteomics) or massive sequencing technologies (transcript analysis). The low amount of starting tissue, its cost-efficiency compared with the utilization of commercial kits, and its performance over a wide range of plant, microbial, and algal species such as Chlamydomonas, Arabidopsis, Populus, or Pinus, makes this method a universal alternative for multiple molecular isolation from plant tissues.
- MeSH
- Arabidopsis genetika metabolismus MeSH
- borovice genetika metabolismus MeSH
- Chlamydomonas reinhardtii genetika metabolismus MeSH
- DNA rostlinná izolace a purifikace MeSH
- genomika metody MeSH
- metabolomika metody MeSH
- Populus genetika metabolismus MeSH
- proteomika metody MeSH
- reprodukovatelnost výsledků MeSH
- RNA rostlin izolace a purifikace MeSH
- rostlinné proteiny izolace a purifikace MeSH
- rostliny * genetika metabolismus MeSH
- systémová biologie metody MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cork oak (Quercus suber L.) is a research priority in the Mediterranean area and because of cork oaks' distribution these stands are experiencing daily stress. Based on projections of intensifying climate change and considering the key role of exploring the recovery abilities, cork oak seedlings were subjected to a cumulative temperature increase from 25°C to 55°C and subsequent recovery. CO2 assimilation rate, chlorophyll fluorescence, anthocyanins, proline and lipid peroxidation were used to evaluate plant performance, while the relative abundance of seven genes encoding for proteins of cork oak with a putative role in thermal/stress regulation (POX1, POX2, HSP10.4, HSP17a.22, CHS, MTL and RBC) was analyzed by qPCR (quantitative Polymerase Chain Reaction). A temperature change to 35°C showed abundance alterations in the tested genes; at 45°C, the molecular changes were associated with an antioxidant response, possibly modulated by anthocyanins. At 55°C, HSP17a.22, MTL and proline accumulation were evident. After recovery, physiological balance was restored, whereas POX1, HSP10.4 and MTL abundances were suggested to be involved in increased thermotolerance. The data presented here are expected to pinpoint some pathways changes occurring during such stress and further recovery in this particular Mediterranean species.
- MeSH
- anthokyaniny metabolismus MeSH
- antioxidancia metabolismus MeSH
- chlorofyl metabolismus MeSH
- dub (rod) genetika fyziologie MeSH
- fotosyntéza MeSH
- fyziologický stres * MeSH
- komplementární DNA genetika MeSH
- oxid uhličitý analýza metabolismus MeSH
- peroxidace lipidů MeSH
- prolin metabolismus MeSH
- regulace genové exprese u rostlin * MeSH
- RNA rostlin genetika MeSH
- rostlinné proteiny genetika MeSH
- vysoká teplota MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
