Abstract

BackgroundSugarcane is a subtropical crop that produces large amounts of biomass annually. It is a key agricultural crop in many countries for the production of sugar and other products. Residual bagasse following sucrose extraction is currently underutilized and it has potential as a carbohydrate source for the production of biofuels. As with all lignocellulosic crops, lignin acts as a barrier to accessing the polysaccharides, and as such, is the focus of transgenic efforts. In this study, we used RNAi to individually reduce the expression of three key genes in the lignin biosynthetic pathway in sugarcane. These genes, caffeoyl-CoA O-methyltransferase (CCoAOMT), ferulate 5-hydroxylase (F5H) and caffeic acid O-methyltransferase (COMT), impact lignin content and/or composition.ResultsFor each RNAi construct, we selected three events for further analysis based on qRT-PCR results. For the CCoAOMT lines, there were no lines with a reduction in lignin content and only one line showed improved glucose release. For F5H, no lines had reduced lignin, but one line had a significant increase in glucose release. For COMT, one line had reduced lignin content, and this line and another released higher levels of glucose during enzymatic hydrolysis. Two of the lines with improved glucose release (F5H-2 and COMT-2) also had reduced S:G ratios.ConclusionsAlong with improvements in bagasse quality for the production of lignocellulosic-based fuels, there was only one line with reduction in juice sucrose extraction, and three lines with significantly improved sucrose production, providing evidence that the alteration of sugarcane for improved lignocellulosic ethanol production can be achieved without negatively impacting sugar production and perhaps even enhancing it.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-016-0683-y) contains supplementary material, which is available to authorized users.

Highlights

  • Sugarcane is a subtropical crop that produces large amounts of biomass annually

  • Generation of RNA interference (RNAi) constructs The sequence for caffeic acid O-methyltransferase (COMT) was available (AY365419; AJ231133), and caffeoyl-CoA O-methyltransferase (CCoAOMT) and ferulate 5-hydroxylase (F5H) were generated from available sugarcane expressed sequence tags (EST) databases (CCoAOMT: CA168805, CA071322, CA159865, CA180815, CA159865, CF575000, CA279207, CA179873; F5H: CA185931, CA134666, CA135938, CA287472, CA278023, CA253395, CA103877)

  • CCoAOMT and F5H RNAi plants both showed the greatest reduction of targeted gene expression in maturing tissue with little to no downregulation observed in leaf or young internodes

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Summary

Introduction

Sugarcane is a subtropical crop that produces large amounts of biomass annually It is a key agricultural crop in many countries for the production of sugar and other products. We used RNAi to individually reduce the expression of three key genes in the lignin biosynthetic pathway in sugarcane. After the extraction of the juice for which it is grown, the remaining plant material is often used inefficiently for the production of energy through burning. This lignocellulosic material could be used for the production of biofuels, adding value to an existing commodity [1]. Previous work has shown that the downregulation of CCoAOMT results in decreases in G monomers

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