A knockout mutation in the lignin biosynthesis gene CCR1 explains a major QTL for acid detergent lignin content in Brassica napus seeds

Brassica napus meal contains high levels of lignin, which is one of the most important compositional factors affecting feed utilization by ruminants. We attempted to modify the concentration and composition of lignin in B. napus plants using the RNAi approach. Four genes were targeted for silencing by this approach either independently or in combination; caffeic acid O-methyltransferase (COMT), cinnamate 4-hydroxylase (C4H); coumarate 3-hydroxylase (C3H); ferulic acid 5-hydroxylase (F5H). We successfully developed transgenic B. napus lines expressing CaMV35S:C3H-C4H RNAi, CaMV35S:F5H-COMT RNAi, and Cruciferin:COMT RNAi that contained up to 40% less seed lignin in the transgenic seeds compared to the control. Despite successfully achieving suppression of these lignin biosynthesis genes and reduction in lignin content in B. napus seeds, we observed minor phenotypic effects on the transgenic plants. In lines carrying the cruciferin:COMT RNAi construct we observed a decrease in lignin content (40%) in the seed and anatomical variations when stem sections were examined. While our silencing had no major negative effect on plant growth it resulted in deformation of vessel elements, and minor changes in S-units. Taken together, these results clearly show that by employing RNAi strategy, it is possible to alter seed lignin content and composition in a manner non-detrimental to B. napus plants.Key words: Brassica napus, cruciferin, lignin, COMT, RNAi

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A major QTL on chromosome C05 significantly reduces acid detergent lignin (ADL) content and increases seed oil and protein content in oilseed rape (Brassica napus L.)

Theoretical and Applied Genetics ◽

10.1007/s00122-018-3167-6 ◽

2018 ◽

Vol 131 (11) ◽

pp. 2477-2492 ◽

Cited By ~ 5

Author(s):

Nina Behnke ◽

Edy Suprianto ◽

Christian Möllers

Keyword(s):

Brassica Napus ◽

Protein Content ◽

Oilseed Rape ◽

Seed Oil ◽

Brassica Napus L ◽

Acid Detergent Lignin ◽

Major Qtl

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Overexpression of Cinnamoyl-CoA Reductase 2 in Brassica napus Increases Resistance to Sclerotinia sclerotiorum by Affecting Lignin Biosynthesis

Frontiers in Plant Science ◽

10.3389/fpls.2021.732733 ◽

2021 ◽

Vol 12 ◽

Author(s):

Dongxiao Liu ◽

Jian Wu ◽

Li Lin ◽

Panpan Li ◽

Saifen Li ◽

...

Keyword(s):

Brassica Napus ◽

Sclerotinia Sclerotiorum ◽

Lignin Content ◽

Lignin Biosynthesis ◽

Normal Growth ◽

Economic Losses ◽

Post Inoculation ◽

Infection Period ◽

Cinnamoyl Coa Reductase ◽

Coa Reductase

Sclerotinia sclerotiorum causes severe yield and economic losses for many crop and vegetable species, especially Brassica napus. To date, no immune B. napus germplasm has been identified, giving rise to a major challenge in the breeding of Sclerotinia resistance. In the present study, we found that, compared with a Sclerotinia-susceptible line (J902), a Sclerotinia-resistant line (J964) exhibited better xylem development and a higher lignin content in the stems, which may limit the invasion and spread of S. sclerotiorum during the early infection period. In addition, genes involved in lignin biosynthesis were induced under S. sclerotiorum infection in both lines, indicating that lignin was deposited proactively in infected tissues. We then overexpressed BnaC.CCR2.b, which encodes the first rate-limiting enzyme (cinnamoyl-CoA reductase) that catalyzes the reaction of lignin-specific pathways, and found that overexpression of BnaC.CCR2.b increased the lignin content in the stems of B. napus by 2.28–2.76% under normal growth conditions. We further evaluated the Sclerotinia resistance of BnaC.CCR2.b overexpression lines at the flower-termination stage and found that the disease lesions on the stems of plants in the T2 and T3 generations decreased by 12.2–33.7% and 32.5–37.3% compared to non-transgenic control plants, respectively, at 7days post-inoculation (dpi). The above results indicate that overexpression of BnaC.CCR2.b leads to an increase in lignin content in the stems, which subsequently leads to increased resistance to S. sclerotiorum. Our findings demonstrate that increasing the lignin content in the stems of B. napus is an important strategy for controlling Sclerotinia.

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Targeting hydroxycinnamoyl CoA: shikimate hydroxycinnamoyl transferase for lignin modification in Brachypodium distachyon

Biotechnology for Biofuels ◽

10.1186/s13068-021-01905-1 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Juan Carlos Serrani-Yarce ◽

Luis Escamilla-Trevino ◽

Jaime Barros ◽

Lina Gallego-Giraldo ◽

Yunqiao Pu ◽

...

Keyword(s):

Negative Impact ◽

Lignin Content ◽

Brachypodium Distachyon ◽

Lignin Biosynthesis ◽

Reverse Direction ◽

Wall Structure ◽

Kinetic Properties ◽

Loss Of Function ◽

Down Regulation ◽

Lignin Modification

Abstract Background Hydroxycinnamoyl CoA: shikimate hydroxycinnamoyl transferase (HCT) is a central enzyme of the so-called “esters” pathway to monolignols. As originally envisioned, HCT functions twice in this pathway, to form coumaroyl shikimate and then, in the “reverse” direction, to convert caffeoyl shikimate to caffeoyl CoA. The discovery of a caffeoyl shikimate esterase (CSE) that forms caffeic acid directly from caffeoyl shikimate calls into question the need for the reverse HCT reaction in lignin biosynthesis. Loss of function of HCT gives severe growth phenotypes in several dicot plants, but less so in some monocots, questioning whether this enzyme, and therefore the shikimate shunt, plays the same role in both monocots and dicots. The model grass Brachypodium distachyon has two HCT genes, but lacks a classical CSE gene. This study was therefore conducted to evaluate the utility of HCT as a target for lignin modification in a species with an “incomplete” shikimate shunt. Results The kinetic properties of recombinant B. distachyon HCTs were compared with those from Arabidopsis thaliana, Medicago truncatula, and Panicum virgatum (switchgrass) for both the forward and reverse reactions. Along with two M. truncatula HCTs, B. distachyon HCT2 had the least kinetically unfavorable reverse HCT reaction, and this enzyme is induced when HCT1 is down-regulated. Down regulation of B. distachyon HCT1, or co-down-regulation of HCT1 and HCT2, by RNA interference led to reduced lignin levels, with only modest changes in lignin composition and molecular weight. Conclusions Down-regulation of HCT1, or co-down-regulation of both HCT genes, in B. distachyon results in less extensive changes in lignin content/composition and cell wall structure than observed following HCT down-regulation in dicots, with little negative impact on biomass yield. Nevertheless, HCT down-regulation leads to significant improvements in biomass saccharification efficiency, making this gene a preferred target for biotechnological improvement of grasses for bioprocessing.

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A High-Density Genetic Map Identifies a Novel Major QTL for Boron Efficiency in Oilseed Rape (Brassica napus L.)

PLoS ONE ◽

10.1371/journal.pone.0112089 ◽

2014 ◽

Vol 9 (11) ◽

pp. e112089 ◽

Cited By ~ 34

Author(s):

Didi Zhang ◽

Yingpeng Hua ◽

Xiaohua Wang ◽

Hua Zhao ◽

Lei Shi ◽

...

Keyword(s):

Brassica Napus ◽

Genetic Map ◽

Oilseed Rape ◽

High Density ◽

Brassica Napus L ◽

Boron Efficiency ◽

Major Qtl

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Effects of the Harvest Stage of Maize Hybrids on the Chemical Composition of Plant Fractions: An Analysis of the Different Types of Silage

Agriculture ◽

10.3390/agriculture11080786 ◽

2021 ◽

Vol 11 (8) ◽

pp. 786

Author(s):

Egon Henrique Horst ◽

Valter Harry Bumbieris Junior ◽

Mikael Neumann ◽

Secundino López

Keyword(s):

Chemical Composition ◽

Lignin Content ◽

Neutral Detergent Fiber ◽

Maturity Stage ◽

Important Distinction ◽

Maize Hybrids ◽

Acid Detergent Lignin ◽

Whole Plant ◽

Grain Formation ◽

Early Harvest

The chemical composition of plant components of three maize hybrids harvested at the beginning of six reproductive stages of maturity was compared. The hybrids evaluated included Maximus VIP3, Defender VIP and Feroz VIP, which were evaluated at each of following stages: R1 (grain formation), R2 (milky grain), R3 (pasty grain), R4 (floury grain), R5 (hard grain) and R6 (ripe grain). The advancement in maturation was linearly related to the crude protein (CP) content of the stem, whole plant, and leaves, and there was a difference among the hybrids. Between R4 and R5 stages, Maximus and Defender presented the highest CP contents for husk (6.58 and 5.42% for Maximus; 5.54 and 5.17% for Defender). The neutral detergent fiber (NDF) of the leaves showed a quadratic relationship with the advancement of maturation but did not differ among the hybrids. For all the hybrids, the NDF content in the husk and cobs increased linearly during the reproductive stages (>77 and 78%, respectively, for the three hybrids in R6). Defender had the lowest NDF content of the cob in R3. The acid detergent lignin contents did not differ among stages in the stems, and showed a linear decrease throughout the whole plant, though the contents did not differ among the hybrids. Due to the differences observed, recommendations for harvest based on the maturity stage for each hybrid should be taken into consideration. There seems to be no important distinction among hybrids for harvesting and use of straw. Despite the reduction in grain yield, an early harvest for earlage or snaplage can provide lower lignin content in husk and cob, as well as higher protein content in the husk, favoring the nutritional value of the vegetative fraction (husk and/or cob).

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Distinct and overlapping functions of Miscanthus sinensis MYB transcription factors SCM1 and MYB103 in lignin biosynthesis

10.1101/629709 ◽

2019 ◽

Author(s):

Philippe Golfier ◽

Faride Unda ◽

Emily K. Murphy ◽

Jianbo Xie ◽

Feng He ◽

...

Keyword(s):

Cell Wall ◽

Transcriptional Regulation ◽

Secondary Cell Wall ◽

Lignin Content ◽

Lignin Biosynthesis ◽

Miscanthus Sinensis ◽

Cell Wall Formation ◽

Transcriptional Responses ◽

Secondary Cell Wall Formation ◽

Wall Formation

AbstractCell wall recalcitrance is a major constraint for the exploitation of lignocellulosic biomass as renewable resource for energy and bio-based products. Transcriptional regulators of the lignin biosynthetic pathway represent promising targets for tailoring lignin content and composition in plant secondary cell walls. A wealth of research in model organisms has revealed that transcriptional regulation of secondary cell wall formation is orchestrated by a hierarchical transcription factor (TF) network with NAC TFs as master regulators and MYB factors in the lower tier regulators. However, knowledge about the transcriptional regulation of lignin biosynthesis in lignocellulosic feedstocks, such as Miscanthus, is limited. Here, we characterized two Miscanthus MYB TFs, MsSCM1 and MsMYB103, and compared their transcriptional impact with that of the master regulator MsSND1. In Miscanthus leaves MsSCM1 and MsMYB103 are expressed at growth stages associated with lignification. Ectopic expression of MsSCM1 and MsMYB103 in tobacco leaves was sufficient to trigger secondary cell wall deposition with distinct sugar and lignin composition. Moreover, RNA-seq analysis revealed that the transcriptional responses to MsSCM1 and MsMYB103 overexpression showed extensive overlap with the response to MsSND1, but were distinct from each other, underscoring the inherent complexity of secondary cell wall formation. Together, MsSCM1 and MsMYB103 represent interesting targets for manipulations of lignin content and composition in Miscanthus towards tailored biomass.

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Gains achieved by molecular approaches in the area of lignification

Pure and Applied Chemistry ◽

10.1351/pac200173030561 ◽

2001 ◽

Vol 73 (3) ◽

pp. 561-566 ◽

Cited By ~ 5

Author(s):

Alain-M. Boudet ◽

Matthieu Chabannes

Keyword(s):

Molecular Biology ◽

Lignin Content ◽

Ectopic Expression ◽

Lignin Biosynthesis ◽

Cell Types ◽

Molecular Approaches ◽

Regulatory Circuits ◽

Recent Developments ◽

Spatio Temporal ◽

Different Cell Types

In this article we highlight the contribution of molecular biology and lignin genetic engineering toward a better understanding of lignin biosynthesis and spatio-temporal deposition of lignin. Specific examples from the literature and from our laboratory will serve to underline the chemical flexibility of lignins, the complexity of the regulatory circuits involved in their synthesis, and the specific behavior of different cell types within the xylem. We will also focus on strategies aiming to reduce the lignin content or to modify the lignin composition of plants and present their impact on plant development. We will show that the ectopic expression of a specific transgene may have a different impact, depending on the genetic background, and that plants with a severe reduction in lignin content may undergo normal development. Lignification is currently benefiting enormously from recent developments in molecular biology and transgenesis, and the progress made opens the way for future developments to study how the walls of lignified plant cells are built and organized.

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Patatin-Related Phospholipase AtpPLAIIIα Affects Lignification of Xylem in Arabidopsis and Hybrid Poplars

Plants ◽

10.3390/plants9040451 ◽

2020 ◽

Vol 9 (4) ◽

pp. 451 ◽

Cited By ~ 1

Author(s):

Jin Hoon Jang ◽

Ok Ran Lee

Keyword(s):

Cell Walls ◽

Lignin Content ◽

Lignin Biosynthesis ◽

Biofuel Production ◽

Amide Bonds ◽

Hybrid Poplars ◽

Plant Lipids ◽

Lipid Acyl Hydrolase ◽

Specific Regulation

Lipid acyl hydrolase are a diverse group of enzymes that hydrolyze the ester or amide bonds of fatty acid in plant lipids. Patatin-related phospholipase AIIIs (pPLAIIIs) are one of major lipid acyl hydrolases that are less closely related to potato tuber patatins and are plant-specific. Recently, overexpression of ginseng-derived PgpPLAIIIβ was reported to be involved in the reduced level of lignin content in Arabidopsis and the mature xylem layer of poplar. The presence of lignin-polysaccharides renders cell walls recalcitrant for pulping and biofuel production. The tissue-specific regulation of lignin biosynthesis, without altering all xylem in plants, can be utilized usefully by keeping mechanical strength and resistance to various environmental stimuli. To identify another pPLAIII homolog from Arabidopsis, constitutively overexpressed AtpPLAIIIα was characterized for xylem lignification in two well-studied model plants, Arabidopsis and poplar. The characterization of gene function in annual and perennial plants with respect to lignin biosynthesis revealed the functional redundancy of less lignification via downregulation of lignin biosynthesis-related genes.

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