Downregulation of Cinnamyl Alcohol Dehydrogenase (CAD) Leads to Improved Saccharification Efficiency in Switchgrass

Keshwani DR, Cheng JJ (2009) Switchgrass for bioethanol and other value-added applications: a review. Bioresour Technol 100:1515–1523

McLaughlin SB, Kszos LA (2005) Development of switchgrass (Panicum virgatum) as a bioenergy feedstock in the United States. Biomass Bioenergy 28:515–535

Bouton JH (2007) Molecular breeding of switchgrass for use as a biofuel crop. Curr Opin Genet Dev 17:553–558

Schmer MR, Vogel KP, Mitchell RB, Perrin RK (2008) Net energy of cellulosic ethanol from switchgrass. Proc Natl Acad Sci USA 105:464–469

Dien BS, Jung H-JG, Vogel KP, Casler MD, Lamb JFS, Iten L et al (2006) Chemical composition and response to dilute-acid pretreatment and enzymatic saccharification of alfalfa, reed canarygrass, and switchgrass. Biomass Bioenergy 30:880–891

Chen F, Dixon RA (2007) Lignin modification improves fermentable sugar yields for biofuel production. Nat Biotechnol 25:759–761

Chapple C, Ladisch M, Meilan R (2007) Loosening lignin’s grip on biofuel production. Nat Biotechnol 25:746–748

Sticklen MB (2008) Plant genetic engineering for biofuel production: towards affordable cellulosic ethanol. Nat Rev Genet 9:433–443

Ralph J, Akiyama T, Kim H, Lu F, Schatz PF, Marita JM et al (2006) Effects of coumarate 3-hydroxylase down-regulation on lignin structure. J Biol Chem 281:8843–8853

Hisano H, Nandakumar R, Wang Z-Y (2009) Genetic modification of lignin biosynthesis for improved biofuel production. In Vitro Cell Dev Biol Plant 45:306–313

Rogers LA, Campbell MMC (2004) The genetic control of lignin deposition during plant growth and development. New Phytol 164:30

Baucher M, Monties B, Van Montagu M, Boerjan W (1998) Biosynthesis and genetic engineering of lignin. Crit Rev Plant Sci 17:125–197

Bernard-Vailhe MA, Besle JM, Maillot MP, Cornu A, Halpin C, Knight M (1998) Effect of down-regulation of cinnamyl alcohol dehydrogenase on cell wall composition and on degradability of tobacco stems. J Sci Food Agric 76:505–514

Yahiaoui N, Marque C, Myton KE, Negrel J, Boudet AM (1998) Impact of different levels of cinnamyl alcohol dehydrogenase down-regulation on lignins of transgenic tobacco plants. Planta 204:8–15

Baucher M, Bernard Vailhe MA, Chabbert B, Besle JM, Opsomer C, Van Montagu M et al (1999) Down-regulation of cinnamyl alcohol dehydrogenase in transgenic alfalfa (Medicago sativa L.) and the effect on lignin composition and digestibility. Plant Mol Biol 39:437–447

Jackson L, Shadle G, Zhou R, Nakashima J, Chen F, Dixon R (2008) Improving saccharification efficiency of alfalfa stems through modification of the terminal stages of monolignol biosynthesis. Bioenerg Res 1:180–192

Baucher M, Chabbert B, Pilate G, Tollier MT, Petit Conil M, Cornu D et al (1996) Red xylem and higher lignin extractability by down-regulating a cinnamyl alcohol dehydrogenase in poplar. Plant Physiol 112:1479–1490

Lapierre C, Pollet B, Petit Conil M, Toval G, Romero J, Pilate G et al (1999) Structural alterations of lignins in transgenic poplars with depressed cinnamyl alcohol dehydrogenase or caffeic acid O-methyltransferase activity have an opposite impact on the efficiency of industrial kraft pulping. Plant Physiol 119:153–163

Pilate G, Guiney E, Holt K, PetitConil M, Lapierre C, Leple JC et al (2002) Field and pulping performances of transgenic trees with altered lignification. Nat Biotechnol 20:607–612

Valério L, Carter D, Rodrigues JC, Tournier V, Gominho J, Marque C et al (2003) Down regulation of cinnamyl alcohol dehydrogenase, a lignification enzyme, in Eucalyptus camaldulensis. Mol Breed 12:157–167

Wróbel-Kwiatkowska M, Starzycki M, Zebrowski J, Oszmianski J, Szopa J (2007) Lignin deficiency in transgenic flax resulted in plants with improved mechanical properties. J Biotechnol 128:919–934

Chen L, Auh C, Dowling P, Bell J, Chen F, Hopkins A et al (2003) Improved forage digestibility of tall fescue (Festuca arundinacea) by transgenic down-regulation of cinnamyl alcohol dehydrogenase. Plant Biotechnol J 1:437–449

Sattler SE, Saathoff AJ, Haas EJ, Palmer NA, Funnell-Harris DL, Sarath G et al (2009) A nonsense mutation in a cinnamyl alcohol dehydrogenase gene is responsible for the sorghum brown midrib6 phenotype. Plant Physiol 150:584–595

Saballos A, Ejeta G, Sanchez E, Kang C, Vermerris W (2009) A genomewide analysis of the cinnamyl alcohol dehydrogenase family in sorghum [Sorghum bicolor (L.) Moench] identifies SbCAD2 as the brown midrib6 gene. Genetics 181:783–795

Halpin C, Holt K, Chojecki J, Oliver D, Chabbert B, Monties B et al (1998) Brown-midrib maize (bm1) - a mutation affecting the cinnamyl alcohol dehydrogenase gene. Plant J 14:545–553

Wang Z-Y, Ge Y (2006) Recent advances in genetic transformation of forage and turf grasses. In Vitro Cell Dev Biol Plant 42:1–18

Moore KJ, Moser LE, Vogel KP, Waller SS, Johnson BE, Pedersen JF (1991) Describing and quantifying growth stages of perennial forage grasses. Agron J 83:1073–1077

Kumar S, Tamura K, Nei M (2004) MEGA3: Integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5:150–163

Miki D, Shimamoto K (2004) Simple RNAi vectors for stable and transient suppression of gene function in rice. Plant Cell Physiol 45:490–495

Xi Y, Fu C, Ge Y, Nandakumar R, Hisano H, Bouton J et al (2009) Agrobacterium-mediated transformation of switchgrass and inheritance of the transgenes. Bioenerg Res 2:275–283

Ramakers C, Ruijter JM, Deprez RHL, Moorman AFM (2003) Assumption-free analysis of quantitative real-time polymerase chain reaction (PCR) data. Neurosci Lett 339:62–66

Ranasinghe S, Rogers ME, Hamilton JGC, Bates PA, Maingon RDC (2008) A real-time PCR assay to estimate Leishmania chagasi load in its natural sand fly vector Lutzomyia longipalpis. Trans R Soc Trop Med Hyg 102:875–882

Zhang K, Qian Q, Huang Z, Wang Y, Li M, Hong L et al (2006) GOLD HULL AND INTERNODE2 encodes a primarily multifunctional cinnamyl-alcohol dehydrogenase in rice. Plant Physiol 140:972–983

Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

dos Santos WD, MdLL F, Ferrarese-Filho O (2006) High performance liquid chromatography method for the determination of cinnamyl alcohol dehydrogenase activity in soybean roots. Plant Physiol Biochem 44:511–515

Chen L, Auh C, Chen F, Cheng XF, Aljoe H, Dixon RA et al (2002) Lignin deposition and associated changes in anatomy, enzyme activity, gene expression and ruminal degradability in stems of tall fescue at different developmental stages. J Agric Food Chem 50:5558–5565

Hatfield RD, Grabber J, Ralph J, Brei K (1999) Using the acetyl bromide assay to determine lignin concentrations in herbaceous plants: some cautionary notes. J Agric Food Chem 47:628–632

Lapierre C, Pollet B, Rolando C (1995) New insight into the molecular architecture of hardwood lignins by chemical degradative method. Res Chem Intermed 21:397–412

Sarath G, Baird LM, Vogel KP, Mitchell RB (2007) Internode structure and cell wall composition in maturing tillers of switchgrass (Panicum virgatum L.). Bioresour Technol 98:2985–2992

Shen H, Fu C, Xiao X, Ray T, Tang Y, Wang Z et al (2009) Developmental control of lignification in stems of lowland switchgrass variety Alamo and the effects on saccharification efficiency. Bioenerg Res 2:233–245

Huhman DV, Berhow MA, Sumner LW (2005) Quantification of saponins in aerial and subterranean tissues of Medicago truncatula. J Agric Food Chem 53:1914–1920

Ghose TK (1987) Measurement of cellulase activities. Pure Appl Chem 59:257–268

Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356

Kim S-J, Kim M-R, Bedgar DL, Moinuddin SGA, Cardenas CL, Davin LB et al (2004) Functional reclassification of the putative cinnamyl alcohol dehydrogenase multigene family in Arabidopsis. Proc Natl Acad Sci USA 101:1455–1460

Saathoff AJ, Tobias CM, Sattler SE, Haas EJ, Twigg P, Sarath G (2010) Switchgrass contains two cinnamyl alcohol dehydrogenases involved in lignin formation. Bioenerg Res. doi: 10.1007/s12155-010-9106-2

Bernard-Vailhe MA, Cornu A, Robert D, Maillot MP, Besle JM (1996) Cell wall degradability of transgenic tobacco stems in relation to their chemical extraction and lignin quality. J Agric Food Chem 44:1164–1169

Palmer N, Sattler S, Saathoff A, Funnell D, Pedersen J, Sarath G (2008) Genetic background impacts soluble and cell wall-bound aromatics in brown midrib mutants of sorghum. Planta 229:115–127

Reddy MSS, Chen F, Shadle G, Jackson L, Aljoe H, Dixon RA (2005) Targeted down-regulation of cytochrome P450 enzymes for forage quality improvement in alfalfa (Medicago sativa L.). Proc Natl Acad Sci USA 102:16573–16578

Niggeweg R, Michael AJ, Martin C (2004) Engineering plants with increased levels of the antioxidant chlorogenic acid. Nat Biotechnol 22:746–754

Boudet AM, Kajita S, Grima-Pettenati J, Goffner D (2003) Lignins and lignocellulosics: a better control of synthesis for new and improved uses. Trends Plant Sci 8:576–581