Cell Wall Proteome of Sugarcane Young and Mature Leaves and Stems

Abstract

By characterizing the cell wall proteomes of different sugarcane organs (leaves and stems) at two developmental stages (young vs mature/apical vs basal), it is possible to address unique characteristics in each of them. Four-month-old leaves show a higher proportion of oxido-reductases and proteins related to lipid metabolism (LM), besides a lower proportion of proteins acting on polysaccharides, in comparison to 4-month-old internodes. It is possible to note that sugarcane leaves and young stems have the highest LM rate than all species, which is assumed to be linked to cuticle formation. The data generated enrich the number of cell wall proteins (CWPs) identified in sugarcane, reaching 277. To our knowledge, sugarcane has now the second higher coverage of monocot CWP in plants.Plant biomass is one of the most abundant renewable resources [1] and it has been assumed that plants devote more than 10% of their genome to the biogenesis of cell walls. [2] During plant growth and development, plant cell walls are continuously modified by enzyme action [3] which are important players in the remodeling of cell wall components. The identification of cell wall proteins (CWPs) has been performed in various plant organs mainly in model plants such as Arabidopsis thaliana and Brachypodium distachyon, but also in crops. [4,5] These data could also be used as a source of information to identify possible targets for plant engineering to produce tailored-cell walls more easily amenable to saccharification and production of ethanol of second generation (E2G). [6] In this work, we have established the CWP atlas of leaves and stems of 4-month-old sugarcane plants. DOI: 10.1002/pmic.201700129The aim was to identify genes encoding proteins of interest to produce tailored cell walls allowing more efficient production of E2G from sugarcane residues. Pooling samples from leaves and stems internodes at different developmental stages from sugarcane variety SP80-3280 were used. The samples were named according the classification on the work of Kuijper, 1915 [cited in [7] ] in which growth in length of the internodes and of the leaves are completed at stages 4 and 0 respectively (see Figure 1). Basal internodes (BI) referring to 5-7 internodes, apical internodes (AI) referring to 1-3 internodes, mature leaves (ML) referring to 1-3 leaves, and young leaves (YL) referring to -3, -2, and -1 leaves. Plants were acclimated in a greenhouse at controlled temperature (26°C) with daily irrigation. Internodes and leaves were collected after 4 months of growth and their proteins were extracted.For the extraction of CWPs a nondestructive method was used, based on [8] and adapted by [9] . Three biological replicates were performed. It is important to mention that the infiltration protocol mostly recovers weakly-bound and labile proteins, and it was chosen because of the already proven high efficiency in recovering sugarcane CWPs compared to destructive techniques. [9] Protein identification was performed by MS subsequently and protein digest were f...

Plant Cell Wall Proteomics: A Focus on Monocot Species, Brachypodium distachyon, Saccharum spp. and Oryza sativa

“…is a crop of special economic interest, as it is one of the major sources of sugar and bioethanol. For this crop, ten different plant samples were surveyed, including suspension cells culture, young and mature leaves, and basal and apical internodes at different developmental stages [37,38,39,40,48]. O. sativa was one of the first plant species with a sequenced genome and it is an economically relevant crop worldwide.…”

“…Over the course of time, several other studies have been performed with model plants and crops: Brassica [23,24], Zea mays [25], Cicer arietinum [26], Vitis vinifera [27], Oriza sativa (rice) [28,29,30,31], Nicotiana tabacum [32], Brachypodium distachyon (stiff brome) [33,34,35,36], Saccharum spp. (sugarcane) [37,38,39,40], among others. Due to this functional diversity, CWPs data present differences between monocots and dicots related to the cell wall structure and composition [41], probably due to both metabolisms’ specificities.…”

Cell Wall Proteome of Wheat Grain Endosperm and Outer Layers at Two Key Stages of Early Development

“…To analyze the cell wall proteome, we performed cell wall enrichment and the proteins were extracted with CaCl 2 and LiCl buffers using a protocol successfully used on different plant species and tissues [25,26,28,29,37,41,42]. In this work, we identified a total of 2454 proteins, 734 of them being predicted as secreted proteins present in the cell wall or the apoplast, and called CWPs.…”

“…The identification and functional understanding of these protein actors are essential to gain access to cell wall dynamics. To date, only a few experiments have been reported on grass cell wall proteomes: Brachypodium distachyon [24][25][26], Saccharum spp (sugarcane) [27][28][29], Oryza sativa (rice) [30][31][32][33], and Triticum aestivum (wheat) [34]. Transcriptomic data on the developing wheat grain allowed for the identification of GH genes [15], and recently, we reported the first data on the cell wall proteome of endosperm and the outer layers of wheat grain at a given developmental stage [34].…”

Cell Wall Proteome Investigation of Bread Wheat (Triticum Aestivum) Developing Grain in Endosperm and Outer Layers

“…To date, Arabidopsis thaliana has the best documented with several sub‐proteomes published . Cell wall proteomes of some monocot species like Oryza sativa, Brachypodium distachyon , or Saccharum officinarum have also been investigated but few studies have focused on their grains . Recently, the cell wall proteome of the B. distachyon grain, a model plant for Poaceae , has been published .…”