Lignified Cell Research Articles

Development and diversity of lignin patterns.

Different patterns of lignified cell walls are associated with diverse functions in a variety of plant tissues. These functions rely on the stiffness and hydrophobicity that lignin polymers impart to the cell wall. The precise pattern of subcellular lignin deposition is critical for the structure-function relationship in each lignified cell type. Here, we describe the role of xylem vessels as water pipes, Casparian strips as apoplastic barriers, and the role of asymmetrically lignified endocarp b cells in exploding seed pods. We highlight similarities and differences in the genetic mechanisms underpinning local lignin deposition in these diverse cell types. By bringing together examples from different developmental contexts and different plant species, we propose that comparative approaches can benefit our understanding of lignin patterning mechanisms.

Early Nitrogen Applications Promote the Initiation of Sweetpotato Storage Roots

A number of published studies have focused on the effects of N application timing on sweetpotato yield. However, none of them examined the influence of the timing of N fertilisation on the initiation of storage roots (SRs), which largely determines the number of tubers per plant and substantially affect the yield. A pot experiment was conducted in sand culture to evaluate the influence of N application timing (N applied on the planting day, 3, 7 and 14 days after transplanting (DAT), denoted T1, T3, T7 and T14, respectively, and a no N applied control T0) on the plant acquisition of N and the initiation of SRs of ‘Orleans’ cultivar. A previously-determined optimal N rate for the SR formation (Hoagland solution with 100mg/L N) in the experimental system was used. Rapid SR formation was found to require N during the first week after transplanting. No or late N application delayed the initiation of SRs and reduced the number of SRs. No or delayed N application inhibited the formation of regular vascular cambium (RVC) and anomalous cambium (AC) and led to an increase in lignification of stele cells leading to significantly higher proportion of lignified roots. By contrast, plants supplied with N within 7 DAT had a higher rate of SRs and lower rate of lignified roots. Also, earlier N application promoted total plant and root growth measured as higher biomass and storage root weight, as well as higher N accumulation in plants and higher nitrogen recovery efficiency (NRE). Results suggested that the SR formation required N during the first week after transplanting. Therefore, moderate N should be available in soil before or on planting day.

Advances in Understanding Microbial Deterioration of Buried and Waterlogged Archaeological Woods: A Review

This review provides information on the advances made leading to an understanding of the micromorphological patterns produced during microbial degradation of lignified cell walls of buried and waterlogged archaeological woods. This knowledge not only serves as an important diagnostic signature for identifying the type(s) of microbial attacks present in such woods but also aids in the development of targeted methods for more effective preservation/restoration of wooden objects of historical and cultural importance. In this review, an outline of the chemical and ultrastructural characteristics of wood cell walls is first presented, which serves as a base for understanding the relationship of these characteristics to microbial degradation of lignocellulosic cell walls. The micromorphological patterns of the three different types of microbial attacks—soft rot, bacterial tunnelling and bacterial erosion—reported to be present in waterlogged woods are described. Then, the relevance of understanding microbial decay patterns to the preservation of waterlogged archaeological wooden artifacts is discussed, with a final section proposing research areas for future exploration.

Monolignol export by diffusion down a polymerization-induced concentration gradient.

Lignin, the second most abundant biopolymer, is a promising renewable energy source and chemical feedstock. A key element of lignin biosynthesis is unknown: how do lignin precursors (monolignols) get from inside the cell out to the cell wall where they are polymerized? Modeling indicates that monolignols can passively diffuse through lipid bilayers, but this has not been tested experimentally. We demonstrate significant monolignol diffusion occurs when laccases, which consume monolignols, are present on one side of the membrane. We hypothesize that lignin polymerization could deplete monomers in the wall, creating a concentration gradient driving monolignol diffusion. We developed a two-photon microscopy approach to visualize lignifying Arabidopsis thaliana root cells. Laccase mutants with reduced ability to form lignin polymer in the wall accumulated monolignols inside cells. In contrast, active transport inhibitors did not decrease lignin in the wall and scant intracellular phenolics were observed. Synthetic liposomes were engineered to encapsulate laccases, and monolignols crossed these pure lipid bilayers to form polymer within. A sink-driven diffusion mechanism explains why it has been difficult to identify genes encoding monolignol transporters and why the export of varied phenylpropanoids occurs without specificity. It also highlights an important role for cell wall oxidative enzymes in monolignol export.

Production of reactive oxygen species by PuRBOHF is critical for stone cell development in pear fruit

The production of reactive oxygen species (ROS) by NADPH oxidase, which is also referred to as respiratory burst oxidase homolog (RBOH), affects several processes in plants. However, the role of RBOHs in cell wall lignification is not well understood. In this study, we show that PuRBOHF, an RBOH isoform, plays an important role in secondary wall formation in pear stone cells. ROS were closely associated with lignin deposition and stone cell formation according to microscopy data. In addition, according to the results of an in situ hybridization analysis, the stage-specific expression of PuRBOHF was higher in stone cells than in cells of other flesh tissues. Inhibitors of RBOH activity suppressed ROS accumulation and stone cell lignification in pear fruit. Moreover, transient overexpression of PuRBOHF caused significant changes in the amount of ROS and lignin that accumulated in pear fruit and flesh calli. We further showed that PuMYB169 regulates PuRBOHF expression, while PuRBOHF-derived ROS induces the transcription of PuPOD2 and PuLAC2. The findings of this study indicate that PuRBOHF-mediated ROS production, which is regulated by a lignin-related transcriptional network, is essential for monolignol polymerization and stone cell formation in pear fruit.

A systems genetics approach reveals PbrNSC as a regulator of lignin and cellulose biosynthesis in stone cells of pear fruit

BackgroundStone cells in fruits of pear (Pyrus pyrifolia) negatively influence fruit quality because their lignified cell walls impart a coarse and granular texture to the fruit flesh.ResultsWe generate RNA-seq data from the developing fruits of 206 pear cultivars with a wide range of stone cell contents and use a systems genetics approach to integrate co-expression networks and expression quantitative trait loci (eQTLs) to characterize the regulatory mechanisms controlling lignocellulose formation in the stone cells of pear fruits. Our data with a total of 35,897 expressed genes and 974,404 SNPs support the identification of seven stone cell formation modules and the detection of 139,515 eQTLs for 3229 genes in these modules. Focusing on regulatory factors and using a co-expression network comprising 39 structural genes, we identify PbrNSC as a candidate regulator of stone cell formation. We then verify the function of PbrNSC in regulating lignocellulose formation using both pear fruit and Arabidopsis plants and further show that PbrNSC can transcriptionally activate multiple target genes involved in secondary cell wall formation.ConclusionsThis study generates a large resource for studying stone cell formation and provides insights into gene regulatory networks controlling the formation of stone cell and lignocellulose.

Biochemical response of Sonneratia alba Sm. branches infested by a wood boring moth (Gazi Bay, Kenya).

Infestation by a moth woodborer species is causing mortality of Sonneratia alba Sm. mangrove by tunneling through the inner bark, cambium and conductive tissue. Infestation leads to death of some infested branches, whereas in other cases infested branches have been observed to recover from infestation. We have used Fourier transform infrared spectroscopy (FTIR) to investigate the differences in macromolecule (polysaccharide and lignin) content present in branches that died (D) of the infestation, those that recovered (R) from the infestation and control branches (C) that were not subject to any infestation. Wood samples were taken from four sampling plots (A, B, C and D) in Gazi Bay (Kenya). From each of the four plots, 15 S. alba branches were taken from five trees, from which 1 cm thick discs were cut from each of these branches to be used as samples. To identify the most characteristic FTIR bands for the three groups of samples, Principal Component Analysis (PCA) was applied on the transposed data matrix. Furthermore, canonical discriminant analysis was applied on the data considering the main FTIR band that would be identified through the PCA factors. Finally, One-way ANOVA and post hoc test were used to verify the significance of the observed trends. Branches that recovered from infestation had higher relative abundance of lignified cells. We conclude that insect-infested S. alba undergo changes related to the lignocellulosic contents. The infestation induces a decrease of the proportion of the polysaccharide content and an increase of the proportion of the lignin contents.

Effect of Rhizoctonia spp. on Polyphenol Oxidase Enzyme Activity in Alfalfa Seedling

Soil borne diseases cause significant losses on quantity and quality of many crop species annually. Rhizoctonia is a widespread and ecologically diverse soilborne fungus causing different types of diseases in many plant species including alfalfa (Medicago sativa). Rhizoctonia species have been traditionally identified based on the cell nuclear condition. Polyphenol oxidases (PPOs) are ubiquitous copper-containing enzymes that are widely occurring enzymes among plants. PPOs are involved in the oxidation of polyphenols into quinones (antimicrobial compounds) and lignification of plant cells that contribute to the formation of defense barriers against pathogens. The study was conducted with the aim to determine the effect of indigenous isolates of a multinucleate (Rhizoctonia solani AG-4) and nineteen isolates of binucleate Rhizoctonia on PPO activity in alfalfa (cv. Gea) seedling under in vitro conditions. The activity of PPO enzyme was determined in inoculated and uninoculated control alfalfa plants after ten days from inoculation. There was a significant increase in the activity of PPO after treatment of alfalfa seedling with isolates of Rhizoctonia. Among Rhizoctonia isolates, highest induction of PPO activity was recorded with pathogenic R. solani AG-4. In present study, increased amounts of PPO were also observed in plants that were challenged with Rhizoctonia spp. PPO has a role in catalyzing phenolic oxidation in limiting disease development. PPO may therefore be involved in induction of defense resistance against plant diseases.

Effects of moisture and heat coupling on xylem growth of Pinus tabuliformis in Shenyang, China

Based on the micro-core technology, we can accurately examine the tree cambium phenology and the radial growth at the cellular scale, and reveal the relationship between tree growth and climate. Pinus tabuliformis is one of the constructive species in forests of northern China. We investigated the growth pattern of P. tabuliformis in Shenyang and the changes in cambium and xylem cells throughout the growing season (April to November) in 2020. Results showed that the dividing activity in cambium started in early April and ended at the end of September. Xylem began to grow from the appearance of enlargement cells (mid April) to the end of the disappearance of lignified cells (late October), with a growth trajectory of 'S' shape curve. Approximately 53 xylem cells per row in radical direction were produced in 2020. The maximum growth rate (0.55 ind/row/day) occurred at the end of May, while the change of earlywood and latewood cells occurred at the end of July. When the minimum temperature above 0 ℃ in Shenyang, the cambium began to divide. The minimum critical temperature that affected the beginning and ending of xylem growth was 2-3 ℃. Precipitation promoted the growth in the growing season. The high temperature and insufficient water supply at the end of July were the main factors driving the differentiation of xylem cells to form earlywood and latewood.

Analysis of PRX Gene Family and Its Function on Cell Lignification in Pears (Pyrus bretschneideri)

Class III peroxidases (PRXs) are plant-specific enzymes that play key roles in the responses to biotic and abiotic stress during plant growth and development. In addition, some peroxidases also play roles in plant lignification. In this study, a total of 114 PRX (designated PbPRXs) genes were identified in the pear (Pyrus bretschneideri Rehd) genome based on systematic analysis. These PRX genes were divided into 12 groups based on their phylogenetic relationships. We performed systematic bioinformatics analysis of the PRX genes, including analysis of gene structures, conserved motifs, phylogenetic relationships, and gene expression patterns during pear fruit growth. The PbPRXs are unevenly distributed on the 17 pear chromosomes and some of them on other scaffolds. Gene duplication event analysis indicated that whole-genome duplication (WGD) and segmental duplication play key roles in PRX gene amplification. Ka/Ks analysis suggested that most duplicated PbPRXs experienced purifying selection, with limited functional divergence during the duplication events. Furthermore, the analysis indicated that those highly expressed genes might play significant roles in the lignification of cells to form stone cells in pear fruit. We examined the expression of those highly expressed genes during fruit growth using quantitative real-time PCR (qRT-PCR), verifying differential expression patterns at different stages of fruit. This study provides useful information for further functional analysis of the PRX gene family in pears.

Contribution to the identification of the species Myrcia hatschbachii D. Legrand (Myrtaceae): anatomical and histochemical analyses

The genus Myrcia is used in folk medicine to treat diabetes. The plants used in folk medicine require morphological and anatomical references to attest to its authenticity. This is the first report of the microscopic study of Myrcia hatschbachii. In this sense, the aim of the present study was to describe the anatomical characteristics, in order to contribute to the species' identification. For anatomical and surface analyses free-hand sections were prepared and observed using optical microscopy, simultaneously some materials were processed and observed under scanning electron microscopy. In addition, histochemical tests were performed. The anatomical features described here correspond with previously reported features found in other species of the genus Myrcia. Furthermore, the following anatomical markers were observed in Myrcia hatschbachii: druses in the spongy parenchyma, concave-convex midrib, heart-shaped petiole, oval-shaped stem, C-shaped vascular bundle in the midrib and open arc shape with invaginated ends in the petiole; phenolic compounds in the phloem and lignified cells in the pith. The results of anatomical and histochemical analyses provide micromorphological and microchemical features that can help in the taxonomy and proper identification of the species.

Effects of Fruit Load on Sugar/Acid Quality and Puffiness of Delayed-Harvest Citrus

Delayed harvesting technology is believed to improve the citrus fruit flavor, but improper tree fruit load under delayed harvest might cause puffiness and reduce fruit quality. In order to find out an optimum tree fruit load level to obtain better flavor quality as well as reduce puffiness in delayed-harvest citrus under protected cultivation, experiments were conducted in the present study between 2019 and 2020 to determine the effect of different fruit loads and fruit-bearing per single branch on the soluble sugars and organic acids metabolism in the peel and flesh, the anatomical structure of the matured fruit peel, and fruit texture-related indexes. The results suggested significant negative correlations between leaf N level and flesh sucrose and glucose contents, and between branch P level and flesh citric acid contents; no significant correlation between NPK levels and flesh texture; relatively lower leaf N and branch P under relatively higher load can increase flesh sucrose and glucose accumulation and slow down citric acid degradation to the greater extent, thus optimizing the sugar/acid ratio of fruits during delayed harvest. The lignification of parenchyma cells closely around peel secretory cavities due to ascorbic acid deficiency might be the primary cause for puffiness under low-load treatments.

Transcriptome and metabolomic analysis to reveal the browning spot formation of \u2018Huangguan\u2019 pear

BackgroundBrowning spot (BS) disorders seriously affect the appearance quality of ‘Huangguan’ pear and cause economic losses. Many studies on BS have mainly focused on physiological and biochemical aspects, and the molecular mechanism remains unclear.ResultsIn the present study, the structural characteristics of ‘Huangguan’ pear with BS were observed via scanning electron microscopy (SEM), the water loss and brown spots were evaluated, and transcriptomic and metabolomics analyses were conducted to reveal the molecular mechanism underlying ‘Huangguan’ pear skin browning disorder. The results showed that the occurrence of BS was accompanied by a decrease in the wax layer and an increase in lignified cells. Genes related to wax biosynthesis were downregulated in BS, resulting in a decrease in the wax layer in BS. Genes related to lignin were upregulated at the transcriptional level, resulting in upregulation of metabolites related to phenylpropanoid biosynthesis. Expression of calcium-related genes were upregulated in BS. Cold-induced genes may represent the key genes that induce the formation of BS. In addition, the results demonstrated that exogenous NaH2PO4·2H2O and ABA treatment could inhibit the incidence of BS during harvest and storage time by increasing wax-related genes and calcium-related genes expression and increasing plant resistance, whereas the transcriptomics results indicated that GA3 may accelerate the incidence and index of BS.ConclusionsThe results of this study indicate a molecular mechanism that could explain BS formation and elucidate the effects of different treatments on the incidence and molecular regulation of BS.

Insights Into the Mechanisms Implicated in Pinus pinaster Resistance to Pinewood Nematode.

Pine wilt disease (PWD), caused by the plant–parasitic nematode Bursaphelenchus xylophilus, has become a severe environmental problem in the Iberian Peninsula with devastating effects in Pinus pinaster forests. Despite the high levels of this species' susceptibility, previous studies reported heritable resistance in P. pinaster trees. Understanding the basis of this resistance can be of extreme relevance for future programs aiming at reducing the disease impact on P. pinaster forests. In this study, we highlighted the mechanisms possibly involved in P. pinaster resistance to PWD, by comparing the transcriptional changes between resistant and susceptible plants after infection. Our analysis revealed a higher number of differentially expressed genes (DEGs) in resistant plants (1,916) when compared with susceptible plants (1,226). Resistance to PWN is mediated by the induction of the jasmonic acid (JA) defense pathway, secondary metabolism pathways, lignin synthesis, oxidative stress response genes, and resistance genes. Quantification of the acetyl bromide-soluble lignin confirmed a significant increase of cell wall lignification of stem tissues around the inoculation zone in resistant plants. In addition to less lignified cell walls, susceptibility to the pine wood nematode seems associated with the activation of the salicylic acid (SA) defense pathway at 72 hpi, as revealed by the higher SA levels in the tissues of susceptible plants. Cell wall reinforcement and hormone signaling mechanisms seem therefore essential for a resistance response.

Lignification and cell wall thickening of ray parenchyma cells in Scots pine sapwood

Abstract Scots pine exhibits variations in ray anatomy, which are poorly understood. Some ray parenchyma cells develop thick and lignified cell walls before heartwood formation. We hypothesized that some stands and trees show high numbers of lignified and thick-walled parenchyma cells early in the sapwood. Therefore, a microscopic analysis of Scots pine sapwood from four different stands in Northern Europe was performed on Safranin — Astra blue-stained tangential micro sections from outer and inner sapwood areas. Significant differences in lignification and cell wall thickening of ray parenchyma cells were observed in the outer sapwood between all of the stands for the trees analyzed. On a single tree level, the relative lignification and cell wall thickening of ray parenchyma cells ranged from 4.3% to 74.3% in the outer sapwood. In the inner sapwood, lignification and cell wall thickening of ray parenchyma cells were more frequent. In some trees, however, the difference in lignification and cell wall thickening between inner and outer sapwood was small since early lignification, and cell wall thickening was already more common in the outer sapwood. Ray composition and number of rays per area were not significantly different within the studied material. However, only one Scottish tree had a significantly higher number of ray parenchyma cells per ray. The differences discovered in lignification and cell wall thickening in ray parenchyma cells early in the sapwood of Scots pine are relevant for wood utilization in general and impregnation treatments with protection agents in particular.

Over-immunity mediated abnormal deposition of lignin arrests the normal enlargement of the root tubers of Rehmannia glutinosa under consecutive monoculture stress

The rapid accumulation of lignin in the cell wall is one of important immune defense mechanism in response to adversity stress in plants. In this study, we found that the enlargement of the root tubers of Rehmannia glutinosa (R. glutinosa) is arrested under consecutive monoculture stress, and this process is accompanied by abnormal accumulation of lignin. Meanwhile, the function of key catalytic enzyme genes in lignin biosynthetic pathway under consecutive monoculture stress was systematically analyzed, of which roles of core genes were validated using reverse genetics. We elucidated that an abnormal deposition of lignin in R. glutinosa roots, induced by consecutive monoculture stress, and arrested the enlargement of root tubers. Additionally, by manipulating the key catalytic enzyme gene RgCCR6, we were able to alter lignin content of roots of R. glutinosa, thereby affecting tuber enlargement. We speculate that cell lignification is an important defense strategy in resistance against consecutive monoculture stress, but the overreacted defense hindered the normal enlargement of root tubers. The findings provide new insights for effectively improving yield reductions of root crops subjected to environmental stress.

Growth-defense trade-offs and yield loss in plants with engineered cell walls.

As a major component of plant secondary cell walls, lignin provides structural integrity and rigidity, and contributes to primary defense by providing a physical barrier to pathogen ingress. Genetic modification of lignin biosynthesis has been adopted to reduce the recalcitrance of lignified cell walls to improve biofuel production, tree pulping properties and forage digestibility. However, lignin-modification is often, but unpredictably, associated with dwarf phenotypes. Hypotheses suggested to explain this include: collapsed vessels leading to defects in water and solute transport; accumulation of molecule(s) that are inhibitory to plant growth or deficiency of metabolites that are critical for plant growth; activation of defense pathways linked to cell wall integrity sensing. However, there is still no commonly accepted underlying mechanism for the growth defects. Here, we discuss recent data on transcriptional reprogramming in plants with modified lignin content and their corresponding suppressor mutants, and evaluate growth-defense trade-offs as a factor underlying the growth phenotypes. New approaches will be necessary to estimate how gross changes in transcriptional reprogramming may quantitatively affect growth. Better understanding of the basis for yield drag following cell wall engineering is important for the biotechnological exploitation of plants as factories for fuels and chemicals.

PbCSE1 promotes lignification during stone cell development in pear (Pyrus bretschneideri) fruit

Pear [Pyrus bretschneideri cv. Dangshan Su] fruit quality is not always satisfactory owing to the presence of stone cells, and lignin is the main component of stone cells in pear fruits. Caffeoyl shikimate esterase (CSE) is a key enzyme in the lignin biosynthesis. Although CSE-like genes have been isolated from a variety of plant species, their orthologs are not characterized in pear. In this study, the CSE gene family (PbCSE) from P. bretschneideri was identified. According to the physiological data and quantitative RT-PCR (qRT-PCR), PbCSE1 was associated with lignin deposition and stone cell formation. The overexpression of PbCSE1 increased the lignin content in pear fruits. Relative to wild-type (WT) Arabidopsis, the overexpression of PbCSE1 delayed growth, increased the lignin deposition and lignin content in stems. Simultaneously, the expression of lignin biosynthetic genes were also increased in pear fruits and Arabidopsis. These results demonstrated that PbCSE1 plays an important role in cell lignification and will provide a potential molecular strategy to improve the quality of pear fruits.

CcBLH6, a bell-like homeodomain-containing transcription factor, regulates the fruit lignification pattern.

CcBLH6 is a bell-like homeodomain-containing transcription factor that plays an important role of lignin biosynthesis in the control of fruit lignification pattern in Camellia chekiangoleosa. The fruit of Camellia chekiangoleosa has a unique lignification pattern that features with a thick pericarp containing a low level of lignification. Yet the fruit lignification pattern and the regulatory network of responsible gene transcription are poorly understood. Here, we characterized a bell-like homeodomain-containing (BLH) transcription factor from C. chekiangoleosa, CcBLH6, in the control of fruit lignification. CcBLH6 expression was highly correlated with the unique lignification pattern during fruit development. The ectopic expression of CcBLH6 promoted the lignification process of stem and root in Arabidopsis. We found that expression of genes related to lignin biosynthesis and its transcriptional regulation was altered in transgenic lines. In a Camellia callus-transformation system, overexpression of CcBLH6 greatly enhanced the expression of genes related to lignin biosynthesis and its transcriptional regulation was altered in transgenic lines. In the callus-transformation system, overexpression of CcBLH6 greatly enhanced the lignification of parenchymal cells, and the regulation of several genes involved in lignin accumulation was largely consistent between Arabidopsis and Camellia. Our study reveals a positive role of CcBLH6 in the regulation of lignin biosynthesis during fruit lignification in Camellia.

Subcellular coordination of plant cell wall synthesis.

Organelles of the plant cell cooperate to synthesize and secrete a strong yet flexible polysaccharide-based extracellular matrix: the cell wall. Cell wall composition varies among plant species, across cell types within a plant, within different regions of a single cell wall, and in response to intrinsic or extrinsic signals. This diversity in cell wall makeup is underpinned by common cellular mechanisms for cell wall production. Cellulose synthase complexes function at the plasma membrane and deposit their product into the cell wall. Matrix polysaccharides are synthesized by a multitude of glycosyltransferases in hundreds of mobile Golgi stacks, and an extensive set of vesicle trafficking proteins govern secretion to the cell wall. In this review, we discuss the different subcellular locations at which cell wall synthesis occurs, review the molecular mechanisms that control cell wall biosynthesis, and examine how these are regulated in response to different perturbations to maintain cell wall homeostasis.