Down-regulation of p-coumaroyl quinate/shikimate 3′-hydroxylase (C3′H) and cinnamate 4-hydroxylase (C4H) genes in the lignin biosynthetic pathway of Eucalyptus urophylla × E. grandis leads to improved sugar release

Biotechnology for Biofuels

Robert W. Sykes²  Erica L. Gjersing²  Kirk Foutz¹  William H. Rottmann¹  Sean A. Kuhn¹  Cliff E. Foster⁴  Angela Ziebell²  Geoffrey B. Turner³  Stephen R. Decker³  Maud A. W. Hinchee¹  Mark F. Davis² 

[1] ArborGen Inc., 2011 Broadbank Ct., Ridgeville 29472, SC, USA

[2] National Bioenergy Center, National Renewable Energy Laboratory, 15013 Denver West Parkway, Golden 80401-3393, CO, USA

[3] Biosciences Center, National Renewable Energy Laboratory, 15013 Denver West Parkway, Golden 80401-3393, CO, USA

[4] Great Lakes Bioenergy Research Center, Michigan State University, East Lansing 48824, MI, USA

关键词: Pretreatment;    Lignin biosynthesis;    Genetic modification;    Recalcitrance;    E. grandis;    × ;    Eucalyptus urophylla ;   

Others  :  1228152
DOI  :  10.1186/s13068-015-0316-x

 received in 2015-04-21, accepted in 2015-08-13,  发布年份 2015

【 摘 要 】

Background

Lignocellulosic materials provide an attractive replacement for food-based crops used to produce ethanol. Understanding the interactions within the cell wall is vital to overcome the highly recalcitrant nature of biomass. One factor imparting plant cell wall recalcitrance is lignin, which can be manipulated by making changes in the lignin biosynthetic pathway. In this study, eucalyptus down-regulated in expression of cinnamate 4-hydroxylase (C4H, EC 1.14.13.11) or p-coumaroyl quinate/shikimate 3′-hydroxylase (C3′H, EC 1.14.13.36) were evaluated for cell wall composition and reduced recalcitrance.

Results

Eucalyptus trees with down-regulated C4H or C3′H expression displayed lowered overall lignin content. The control samples had an average of 29.6 %, the C3′H reduced lines had an average of 21.7 %, and the C4H reduced lines had an average of 18.9 % lignin from wet chemical analysis. The C3′H and C4H down-regulated lines had different lignin compositions with average S/G/H ratios of 48.5/33.2/18.3 for the C3′H reduced lines and 59.0/39.8/1.2 for the C4H reduced lines, compared to the control with 65.9/33.2/1.0. Both the C4H and C3′H down-regulated lines had reduced recalcitrance as indicated by increased sugar release as determined using enzymatic conversion assays utilizing both no pretreatment and a hot water pretreatment.

Conclusions

Lowering lignin content rather than altering sinapyl alcohol/coniferyl alcohol/4-coumaryl alcohol ratios was found to have the largest impact on reducing recalcitrance of the transgenic eucalyptus variants. The development of lower recalcitrance trees opens up the possibility of using alternative pretreatment strategies in biomass conversion processes that can reduce processing costs.

【 授权许可】

   
2015 Sykes et al.

【 图 表 】

Fig.1.

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Fig.1.

Fig.2.

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