Upgrading pectin methylation for consistently enhanced biomass enzymatic saccharification and cadmium phytoremediation in rice Ospmes site-mutants.

Crop straws provide enormous biomass residues applicable for biofuel production and trace metal phytoremediation. However, as lignocellulose recalcitrance determines a costly process with potential secondary waste liberation, genetic modification of plant cell walls is deemed as a promising solution. Although pectin methylation plays an important role for plant cell wall construction and integrity, little is known about its regulation roles on lignocellulose hydrolysis and trace metal elimination. In this study, we initially performed a typical CRISPR/Cas9 gene-editing for site mutations of OsPME31, OsPME34 and OsPME79 in rice, and then determined significantly upgraded pectin methylation degrees in the young seedlings of three distinct site-mutants compared to their wild type. We then examined distinctively improved lignocellulose recalcitrance in three mutants including reduced cellulose levels, crystallinity and polymerization or raised hemicellulose deposition and cellulose accessibility, which led to specifically enlarged biomass porosity either for consistently enhanced biomass enzymatic saccharification under mild alkali pretreatments or for cadmium (Cd) accumulation up to 2.4-fold. Therefore, this study proposed a novel model to elucidate how pectin methylation could play a unique enhancement role for both lignocellulose enzymatic hydrolysis and Cd phytoremediation, providing insights into precise pectin modification for effective biomass utilization and efficient trace metal exclusion.

Incomplete genome doubling enables to consistently enhance plant growth for maximum biomass production by altering multiple transcript co-expression networks in potato.

KEY MESSAGE: Cytochimera potato plants, which mixed with diploid and tetraploid cells, could cause the highest and significantly increased biomass yield than the polyploid and diploid potato plants. Polyploidization is an important approach in crop breeding for agronomic trait improvement, especially for biomass production. Cytochimera contains two or more mixed cells with different levels of ploidy, which is considered a failure in whole genome duplication. Using colchicine treatment with diploid (Dip) potato (Solanum chacoense) plantlets, this study generated tetraploid (Tet) and cytochimera (Cyt) lines, which, respectively, contained complete and partial cells with genome duplication. Compared to the Dip potato, we observed remarkably enhanced plant growth and biomass yields in Tet and Cyt lines. Notably, the Cyt potato straw, which was generated from incomplete genome doubling, was of significantly higher biomass yield than that of the Tet with a distinctively altered cell wall composition. Meanwhile, we observed that one layer of the tetraploid cells (about 30%) in Cyt plants was sufficient to trigger a gene expression pattern similar to that of Tet, suggesting that the biomass dominance of Cyt may be related to the proportion of different ploidy cells. Further genome-wide analyses of co-expression networks indicated that down-regulation (against Dip) of spliceosomal-related transcripts might lead to differential alternative splicing for specifically improved agronomic traits such as plant growth, biomass yield, and lignocellulose composition in Tet and Cyt plants. In addition, this work examined that the genome of Cyt line was relatively stable after years of asexual reproduction. Hence, this study has demonstrated that incomplete genome doubling is a promising strategy to maximize biomass production in potatoes and beyond.

Modified lignocellulose and rich starch for complete saccharification to maximize bioethanol in distinct polyploidy potato straw.

Potato is a major food crop with enormous biomass straw, but lignocellulose recalcitrance causes a costly bioethanol conversion. Here, we selected the cytochimera (Cyt) potato samples showing significantly-modified lignocellulose and much increased soluble sugars and starch by 2-4 folds in mature straws. Under two pretreatments (8 min liquid hot water; 5% CaO) at minimized conditions, the potato Cyt straw showed complete enzymatic saccharification. Further performing yeast fermentation with all hexoses released from soluble sugars, starch and lignocellulose in the Cyt straw, this study achieved a maximum bioethanol yield of 24 % (% dry matter), being higher than those of other bioenergy crops as previously reported. Hence, this study has proposed a novel mechanism model on the reduction of major lignocellulose recalcitrance and regulation of carbon assimilation to achieve cost-effective bioethanol production under optimal pretreatments. This work also provides a sustainable strategy for utilization of potato straws with minimum waste release.

Quantum dots are conventionally applicable for wide-profiling of wall polymer distribution and destruction in diverse cells of rice.

Plant cell walls represent enormous biomass resources for biofuels, and it thus becomes important to establish a sensitive and wide-applicable approach to visualize wall polymer distribution and destruction during plant growth and biomass process. Despite quantum dots (QDs) have been applied to label biological specimens, little is reported about its application in plant cell walls. Here, semiconductor QDs (CdSe/ZnS) were employed to label the secondary antibody directed to the epitopes of pectin or xylan, and sorted out the optimal conditions for visualizing two polysaccharides distribution in cell walls of rice stem. Meanwhile, the established QDs approach could simultaneously highlight wall polysaccharides and lignin co-localization in different cell types. Notably, this work demonstrated that the QDs labeling was sensitive to profile distinctive wall polymer destruction between alkali and acid pretreatments with stem tissues of rice. Hence, this study has provided a powerful tool to characterize wall polymer functions in plant growth and development in vivo, as well as their distinct roles during biomass process in vitro.

Miscanthus accessions distinctively accumulate cadmium for largely enhanced biomass enzymatic saccharification by increasing hemicellulose and pectin and reducing cellulose CrI and DP.

In this study, total eight distinct Miscanthus accessions were collected from the cadmium (Cd)-supplied soil pots, and mild alkali pretreatments (0.5%, 1% NaOH) were then performed to enhance biomass enzymatic saccharification. Due to large Cd accumulation, all Miscanthus accessions showed significantly reduced cellulose levels and features (CrI, DP) with much increased hemicellulose and pectin contents in the mature stems. Under mild alkali pretreatments, all Miscanthus samples exhibited largely increased hexoses yields released from enzymatic hydrolysis, and one desirable accession had an almost complete biomass saccharification with the hexoses yield at 100% (% cellulose). Notably, the biomass residues remained from enzymatic hydrolysis upon 1% NaOH pretreatment could absorb 73-96% Cd (% of total), suggesting an applicable approach for Cd phyto-remediation. Hence, a hypothetic model was proposed to elucidate that the enhanced biomass saccharification should be mainly due to much reduced cellulose CrI and DP in the Cd-accumulated Miscanthus accessions.

Silica distinctively affects cell wall features and lignocellulosic saccharification with large enhancement on biomass production in rice.

Rice is a typical silicon-accumulating crop with enormous biomass residues for biofuels. Silica is a cell wall component, but its effect on the plant cell wall and biomass production remains largely unknown. In this study, a systems biology approach was performed using 42 distinct rice cell wall mutants. We found that silica levels are significantly positively correlated with three major wall polymers, indicating that silica is associated with the cell wall network. Silicon-supplied hydroculture analysis demonstrated that silica distinctively affects cell wall composition and major wall polymer features, including cellulose crystallinity (CrI), arabinose substitution degree (reverse Xyl/Ara) of xylans, and sinapyl alcohol (S) proportion in three typical rice mutants. Notably, the silicon supplement exhibited dual effects on biomass enzymatic digestibility in the mutant and wild type (NPB) after pre-treatments with 1% NaOH and 1% H2SO4. In addition, silicon supply largely enhanced plant height, mechanical strength and straw biomass production, suggesting that silica rescues mutant growth defects. Hence, this study provides potential approaches for silicon applications in biomass process and bioenergy rice breeding.