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GCEP-sponsored scientists unmask crucial plant gene that could enhance efficient biofuel production

Science Journal Cover image

Enhanced-color confocal microscopy image of a cross section of an Arabidopsis stem
Cover of Science, click for full report

An international research team has identified a crucial gene that could help scientists overcome a major hurdle in the efficient production of low-cost biofuels from switchgrass, poplar and other environmentally friendly crops.

Writing in the Aug. 15 online edition of the journal Science, the team of scientists from VIB and Ghent University in Belgium, the University of Dundee and The James Hutton Institute in the U.K., and the University of Wisconsin-Madison described a gene with a heretofore-unknown role in the development of lignin, a major component of secondary cell walls in plants. Unmasking this hidden gene, called caffeoyl shikimate esterase (CSE), will "necessitate revision of currently accepted models of the lignin biosynthetic pathway," the authors wrote.

The scientists also discovered that deleting the CSE gene quadruples the amount of sugar that can be extracted from the cell wall - a potentially significant finding for the biofuels industry.

"This exciting, fundamental discovery provides an alternative pathway for altering lignin in plants and has the potential to greatly increase the efficiency of energy crop conversion for biofuels," said Sally M. Benson, director of Stanford University's Global Climate and Energy Project (GCEP), which co-funded the study. "We have been so pleased to support this team of world leaders in lignin research and to see the highly successful outcome of these projects."

Lignin cement

In addition to lignin, plant cell walls also contain large amounts of cellulose, a substance made entirely of the simple sugar, glucose. Lignin acts like a cement that embeds the cellulose in place, giving firmness and rigidity to the plant. The evolutionary emergence of lignin allowed the development of vascular land plants, enabling the transport of water and nutrients from the roots to the leaves, the authors noted. Thanks to lignin, even very tall plants can maintain their upright stature,” they said.

Glucose can be fermented into ethanol, a renewable biofuel, and other alcohols. Most of the glucose used to make ethanol comes from starch- and sugar-based feedstocks, such as edible corn and sugar cane. But using valuable cropland to produce fuel instead of food has raised a number of environmental and social concerns.

As an alternative, researchers have turned to so-called “cellulosic feedstocks,” such as grass and wood, which can be grown on abandoned lands with relatively little water and fertilizer. However, lignin’s tight grip on cellulose has been a significant barrier to the development of biofuels from cellulosic plants. Gaining access to glucose in these plants requires removing the lignin cement that holds the cellulose in place via a pretreatment process that is energy-consuming and environmentally unfriendly.

Genetic approach

In the Science study, the research team proposed a genetic approach that could lead to more efficient processing of cellulosic crops on an industrial scale. Using the model plant Arabidopsis thaliana, the researchers discovered that the CSE gene produces an enzyme with a central role in lignin biosynthesis. Removal of the gene resulted in a smaller plant with 36-percent less lignin per gram of stem material than a normal Arabidopsis, and caused the lignin to loosen its cement-like grip on the cellulose.

As a result, the researchers achieved a four-fold increase in the conversion of cellulose to glucose - a 78-percent conversion rate compared to 18 percent for a normal Arabidopsis plant. This dramatic increase in glucose extraction required no pretreatment of the cell walls, a potentially expensive and caustic process.

Plants with lower amounts of lignin, or with lignin that is easier to break down, can be a real benefit for the biofuel, bioplastics and paper industries, the authors said. As a result of the study, scientists can now screen wild populations of cellulosic energy crops - such as poplar, eucalyptus, switchgrass and other grass species -to see if they carry a mutated or non-functional CSE gene. This screening process could contribute to a more efficient conversion of biomass to energy, they said. Alternatively, scientists might be able to genetically engineer the CSE gene to reduce the amount of lignin in the cell walls of these crops.

The co-lead authors of the study are Ruben Vanholme and Igor Cesarino of VIB and Ghent University. Other authors are from the following institutions:

  • University of Dundee: Katarzyna Rataj, Yuguo Xiao, Lydia Welsh, Christopher McClellan, Claire Halpin and Gordon Simpson (also with The James Hutton Institute);
  • VIB and/or Ghent University: Lisa Sundin, Geert Goeminne, Joanna Cross, Kris Morreel, Pedro Araujo, Jurgen Haustraete, Bartel Vanholme and Wout Boerjan;
  • University of Wisconsin-Madison: Hoon Kim and John Ralph.

The work was supported by the multidisciplinary research partnership, Biotechnology for a Sustainable Economy, at Ghent University; the U.S. Department of Energy Great Lakes Bioenergy Research Center; and the following three awards from GCEP:

The Plant Journal Cover image

Visualization of plant cell wall lignification using fluorescence-tagged monolignols.
Cover of The Plant Journal click for full report

In November 2013, The Plant Journal published a related study on the novel fluorescent-tagging technique developed by the lignin research team.

"A big question in plant biosynthesis is how molecules are transported and combined into polymers," said co-author Yuki Tobimatsu of the University of Wisconsin-Madison. The molecules, called monolignols, are lignin precursors. Tagging monolignols with fluorescent dye allows researchers to follow them through the tissues of living plant seedlings as they are absorbed and utilized, Tobimatsu explained.

"The cool thing about Yuki's dye," added Ralph, "is that it only stains plant tissues that are metabolically active, so only the parts of the plant that are actually producing lignin change color."

The study, featured on the cover of the Journal, was also supported by GCEP.

See also: Lignin Review - Highlights from GCEP Contributions

Compiled from press release from VIB (August 15, 2013) and article from Great Lakes Bienergy Research Center (September 16, 2013).

November 19, 2013



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