Clint Chapple

Rewriting the lignin roadmap

Considerable interest in lignin biosynthesis has been fueled by the many roles that lignin plays in development and in resistance to biotic and abiotic stress, as well as its importance to industry and agriculture. Although the pathway leading to the lignin polymer has been studied for decades, new insights into the enzymes of the pathway have required a complete re-evaluation of how we think lignin precursors are synthesized. Although free hydroxycinnamic acids have long been thought to be key intermediates, it has become apparent that many of the hydroxylation and methylation steps in the pathway occur instead at the level of hydroxycinnamic acid esters, and their corresponding aldehydes and alcohols.

Improvement of biomass through lignin modification.

Lignin, a major component of the cell wall of vascular plants, has long been recognized for its negative impact on forage quality, paper manufacturing, and, more recently, cellulosic biofuel production. Over the last two decades, genetic and biochemical analyses of brown midrib mutants of maize, sorghum and related grasses have advanced our understanding of the relationship between lignification and forage digestibility. This work has also inspired genetic engineering efforts aimed at generating crops with altered lignin, with the expectation that these strategies would enhance forage digestibility and/or pulping efficiency. The knowledge gained from these bioengineering efforts has greatly improved our understanding of the optimal lignin characteristics required for various applications of lignocellulosic materials while also contributing to our understanding of the lignin biosynthetic pathway. The recent upswing of interest in cellulosic biofuel production has become the new focus of lignin engineering. Populus trichocarpa and Brachypodium distachyon are emerging as model systems for energy crops. Lignin research on these systems, as well as on a variety of proposed energy crop species, is expected to shed new light on lignin biosynthesis and its regulation in energy crops, and lead to rational genetic engineering approaches to modify lignin for improved biofuel production.

The origin and evolution of lignin biosynthesis.

SUMMARY Lignin, a phenolic polymer derived mainly from hydroxycinnamyl alcohols, is ubiquitously present in tracheophytes. The development of lignin biosynthesis has been considered to be one of the key factors that allowed land plants to flourish in terrestrial ecosystems. Lignin provides structural rigidity for tracheophytes to stand upright, and strengthens the cell wall of their water-conducting tracheary elements to withstand the negative pressure generated during transpiration. In this review, we discuss a number of aspects regarding the origin and evolution of lignin biosynthesis during land plant evolution, including the establishment of its monomer biosynthetic scaffold, potential precursors to the lignin polymer, as well as the emergence of the polymerization machinery and regulatory system. The accumulated knowledge on the topic, as summarized here, provides us with an evolutionary view on how this complex metabolic system emerged and developed.

The Phenylpropanoid Pathway in Arabidopsis

The phenylpropanoid pathway serves as a rich source of metabolites in plants, being required for the biosynthesis of lignin, and serving as a starting point for the production of many other important compounds, such as the flavonoids, coumarins, and lignans. In spite of the fact that the phenylpropanoids and their derivatives are sometimes classified as secondary metabolites, their relevance to plant survival has been made clear via the study of Arabidopsis and other plant species. As a model system, Arabidopsis has helped to elucidate many details of the phenylpropanoid pathway, its enzymes and intermediates, and the interconnectedness of the pathway with plant metabolism as a whole. These advances in our understanding have been made possible in large part by the relative ease with which mutations can be generated, identified, and studied in Arabidopsis. Herein, we provide an overview of the research progress that has been made in recent years, emphasizing both the genes (and gene families) associated with the phenylpropanoid pathway in Arabidopsis, and the end products that have contributed to the identification of many mutants deficient in the phenylpropanoid metabolism: the sinapate esters.

Loosening lignin's grip on biofuel production.

Reducing lignin content in alfalfa can increase ethanol yields and decrease processing inputs.

Plant cell walls as targets for biotechnology.

Plants are the sources of major food, feed, and fiber products that are used globally. This past year has seen advances in our understanding of the enzymes that modify wall architecture, the cloning of the first cellulose synthase gene, and revisions to the lignin biosynthetic pathway. These discoveries have facilitated the development of new strategies to alter cell wall properties in transgenic plants.

Understanding Lignification: Challenges Beyond Monolignol Biosynthesis

Lignin, a major component of vascular plant cell wall, provides mechanical support for plants to stand upright and enables xylems to withstand the negative pressure generated during water transport. Although important for plant growth, the presence of lignin limits access to cell wall

50 years of Arabidopsis research: highlights and future directions

922 I. 922 II. 922 III. 925 IV. 925 V. 926 VI. 927 VII. 928 VIII. 929 IX. 930 X. 931 XI. 932 XII. 933 XIII. Natural variation and genome-wide association studies 934 XIV. 934 XV. 935 XVI. 936 XVII. 937 937 References 937 SUMMARY: The year 2014 marked the 25(th) International Conference on Arabidopsis Research. In the 50 yr since the first International Conference on Arabidopsis Research, held in 1965 in Göttingen, Germany, > 54 000 papers that mention Arabidopsis thaliana in the title, abstract or keywords have been published. We present herein a citational network analysis of these papers, and touch on some of the important discoveries in plant biology that have been made in this powerful model system, and highlight how these discoveries have then had an impact in crop species. We also look to the future, highlighting some outstanding questions that can be readily addressed in Arabidopsis. Topics that are discussed include Arabidopsis reverse genetic resources, stock centers, databases and online tools, cell biology, development, hormones, plant immunity, signaling in response to abiotic stress, transporters, biosynthesis of cells walls and macromolecules such as starch and lipids, epigenetics and epigenomics, genome-wide association studies and natural variation, gene regulatory networks, modeling and systems biology, and synthetic biology.

36 Secondary Metabolism in Arabidopsis

Plants cannot remove themselves from environments where they may be subjected to various insults, such as attack by pathogens, irradiation by UV light, or damage by herbivores. It is thought that to provide protection against the adverse effects of their environment, plants have acquired the ability to accumulate secondary metabolites. Many of these same compounds are also important to humans because of their pharmaceutical, insecticidal, and organoleptic qualities. Arabidopsis has been reported to accumulate 36 secondary metabolites belonging to four distinct classes. Flavonoids and hydroxycinnamic acid esters are derived from the general phenylpropanoid pathway (Fig. 1). The other two classes of secondary metabolites known to be produced by Arabidopsis are the glucosinolates and the indole phytoalexins. More than 15,000 plant secondary metabolites have been identified, and this may represent only 5–10% of those that occur in nature (Wink 1988). How this diversity has arisen is unknown, as is the extent to which this diversity is represented within a single species, although some hypotheses have been offered (Williams et al. 1989; Jones and Firn 1991). For example, the pathways devoted to cyanogenic glycoside and glucosinolate biosynthesis appear to be closely related, yet they diverge to produce very different secondary metabolites. Cyanogenic glycosides have been found in more than 200 species of angiosperms (Conn 1980) but have never been found in species such as Arabidopsis that accumulate glucosinolates. What has led to this specialization and mutual exclusivity? Are the genes for cyanogenic glycoside biosynthesis present in Arabidopsis, but unexpressed? If so,...

Manipulation Of Lignin Biosynthesis To Maximize Ethanol Production From Populus Feedstocks

Our research focuses on transgenic strategies for modifying lignification to improve biomass quality, without leading to deleterious effects on plant performance. In order to accomplish this objective, we designed molecular strategies and selected appropriate transgenes for manipulating the expression of lignification-associated genes; we generated poplar engineered for altered lignin content and/or monomer composition, and field-tested them for fitness; we analyzed the impact of these transgenic strategies on metabolism in general and lignin biosynthesis in particular; and evaluated the ease with which cell wall deconstruction can be accomplished using both chemical and enzymatic means using wild-type and high syringyl poplar.