Secondary Wall Thickening Research Articles

Stellate Trichomes in Dionaea muscipula Ellis (Venus Flytrap) Traps, Structure and Functions

The digestive organs of carnivorous plants have external (abaxial) glands and trichomes, which perform various functions. Dionaea muscipula Ellis (the Venus flytrap) is a model carnivorous plant species whose traps are covered by external trichomes. The aim of the study was to fill in the gap regarding the structure of the stellate outer trichomes and their immunocytochemistry and to determine whether these data support the suggestions of other authors about the roles of these trichomes. Light and electron microscopy was used to show the trichomes' structure. Fluorescence microscopy was used to locate the carbohydrate epitopes that are associated with the major cell wall polysaccharides and glycoproteins. The endodermal cells and internal head cells of the trichomes were differentiated as transfer cells, and this supports the idea that stellate trichomes transport solutes and are not only tomentose-like trichomes. Trichome cells differ in the composition of their cell walls, e.g., the cell walls of the internal head cells are enriched with arabinogalactan proteins (AGPs). The cell walls of the outer head cells are poor in both low and highly homogalacturonans (HGs), but the immature trichomes are rich in the pectic polysaccharide (1-4)-β-D-galactan. In the immature traps, young stellate trichomes produce mucilage which may protect the trap surface, and in particular, the trap entrance. However, the role of these trichomes is different when the outer head cells collapse. In the internal head cells, a thick secondary wall cell was deposited, which together with the thick cell walls of the outer head cells played the role of a large apoplastic space. This may suggest that mature stellate trichomes might function as hydathodes, but this should be experimentally proven.

Diverse forms of xylem-Like cells and strand formation in Xylogenic Eucalyptus bosistoana callus culture

In vitro xylem induction system is a basic tool in physiological, biochemical, and molecular studies of secondary cell wall formation, lignin biosynthesis and deposition associated with tracheary element formation. Eucalyptus bosistoana is a Class 1 durable hardwood tree species, selected by the New Zealand Dryland Forest Initiative for good quality wood and high adaptability to the NZ growing conditions. Xylogenic E. bosistoana callus culture was established and up to 40% of the callus cells were xylem-like cells (XLCs) which may have differentiated from small, cytoplasmically dense or compact dividing, and exhibited increased lignin contents during culture. The eucalyptus XLCs showed diverse sizes, patterns of secondary cell wall thickenings similar to the xylem cells in the young shoots and organized development including cell–cell connections of the XLCs to form xylem strands. This is the first report of the organized development of XLCs in E. bosistoana callus culture.

FvNST1b NAC Protein Induces Secondary Cell Wall Formation in Strawberry.

Secondary cell wall thickening plays a crucial role in plant growth and development. Diploid woodland strawberry (Fragaria vesca) is an excellent model for studying fruit development, but its molecular control of secondary wall thickening is largely unknown. Previous studies have shown that Arabidopsis NAC secondary wall thickening promoting factor1 (AtNST1) and related proteins are master regulators of xylem fiber cell differentiation in multiple plant species. In this study, a NST1-like gene, FvNST1b, was isolated and characterized from strawberry. Sequence alignment and phylogenetic analysis showed that the FvNST1b protein contains a highly conserved NAC domain, and it belongs to the same family as AtNST1. Overexpression of FvNST1b in wild-type Arabidopsis caused extreme dwarfism, induced ectopic thickening of secondary walls in various tissues, and upregulated the expression of genes related to secondary cell wall synthesis. In addition, transient overexpression of FvNST1b in wild-type Fragaria vesca fruit produced cells resembling tracheary elements. These results suggest that FvNST1b positively regulates secondary cell wall formation as orthologous genes from other species.

The herbaceous peony transcription factor WRKY41a promotes secondary cell wall thickening to enhance stem strength.

Stem bending or lodging caused by insufficient stem strength is an important limiting factor for plant production. Secondary cell walls play a crucial role in plant stem strength, but whether WRKY transcription factors can positively modulate secondary cell wall thickness are remain unknown. Here, we characterized a WRKY transcription factor PlWRKY41a from herbaceous peony (Paeonia lactiflora), which was highly expressed in stems. PlWRKY41a functioned as a nucleus-localized transcriptional activator and enhanced stem strength by positively modulating secondary cell wall thickness. Moreover, PlWRKY41a bound to the promoter of the XYLOGLUCAN ENDOTRANSGLUCOSYLASE/HYDROLASE4 (PlXTH4) and activated the expression of PlXTH4. PlXTH4-overexpressing tobacco (Nicotiana tabacum) had thicker secondary cell walls, resulting in enhanced stem strength, while PlXTH4-silenced P. lactiflora had thinner secondary cell walls, showing decreased stem strength. Additionally, PlWRKY41a directly interacted with PlMYB43 to form a protein complex, and their interaction induced the expression of PlXTH4. These data support that the PlMYB43-PlWRKY41a protein complex can directly activate the expression of PlXTH4 to enhance stem strength by modulating secondary cell wall thickness in P. lactiflora. The results will enhance our understanding of the formation mechanism of stem strength and provide a candidate gene to improve stem straightness in plants.

CEF3 is involved in membrane trafficking and essential for secondary cell wall biosynthesis and its mutation enhanced biomass enzymatic saccharification in rice

BackgroundAs one of the most important staple food crops, rice produces large of agronomic biomass residues that contain lots of secondary cell walls (SCWs). Membrane trafficking plays key roles in SCWs biosynthesis, but information association membrane trafficking and SCWs formation in plants is limited.ResultsIn this study, we report the function characterization of a rice mutant, culm easily fragile 3 (cef3), that exhibits growth retardation and fragile culm phenotype with significantly altered cell wall composition and reduced secondary wall thickness. Map-based cloning revealed that CEF3 encodes a homologous protein of Arabidopsis STOMATAL CYTOKINESIS DEFECTIVE2 (SCD2). The saccharification assays revealed that CEF3 mutation can improve biomass enzymatic saccharification. Expression pattern analysis indicated that CEF3 is ubiquitously expressed in many organs at different developmental stages. Subcellular localization revealed that CEF3 is a Golgi-localized protein. The FM4-64 uptake assay revealed CEF3 is involved in endocytosis. Furthermore, mutation of CEF3 not only affected cellulose synthesis-related genes expression, but also altered the abundance of cellulose synthase catalytic subunit 9 (OsCESA9) in the PM and in the endomembrane systems.ConclusionsThis study has demonstrated that CEF3 participates in the membrane trafficking that is essential for normal cellulose and other polysaccharides biosynthesis of the secondary cell wall, thereby manipulation of CEF3 could alter cellulose content and enhance biomass enzymatic saccharification in rice plants. Therefore, the study of the function of CEF3 can provide a strategy for genetic modification of SCWs in bioenergy crops.

Anatomical adaptions of pits in two types of ray parenchyma cells in Populus tomentosa during the xylem differentiation

Pits in ray parenchyma cells are important to understand the functional anatomy of the ray parenchyma network in the xylem but have been less studied. Herein, pits in two types of ray parenchyma cells, contact cells and isolation cells, across different developmental stages were qualitatively studied using 48-year-old Populus tomentosa trees. The timing of differentiation and death was determined by histochemical staining and polarized light microscopy. The dimension, shape and density of pits as well as cell wall thickness were measured using SEM and optical microscopy images of semi-thin radial sections and macerated ray parenchyma cells, and analyzed by multi-factor analyses of variance. Results showed that secondary wall thickening and lignification of contact cells begun near the cambium, contrarily those of isolation cells have started until the transition zone. But even in the sapwood, contact cell walls were still much thinner than isolation cell walls. Moreover, district anatomical adaptions of pits during the xylem differentiation were present between horizontal walls and tangential walls, between contact cells and isolation cells. Ray pits were simple to slightly bordered, whereas sieve-like pits were only shown on tangential walls of isolation cells. Pit density of horizontal walls was similar between contact cells and isolation cells, nevertheless greater pits were present on tangential walls, especially for isolation cells. In addition, pits of ray parenchyma cells in the heartwood were smaller and more bordered than those in the sapwood, particularly on the horizontal walls. Moreover, isolation cells had pits with the smaller dimensions, greater pits on the tangential walls, more bordered pits on horizontal walls, as well as longer and narrower cell morphology with much thicker cell walls than contact cells. To a certain extent, all these anatomical adaptations were developed to ensure distinct functions of the two types of ray parenchyma cells in the xylem and finally to support tree growth in demand.

High-transcriptional activation ability of bamboo SECONDARY WALL NAC transcription factors is derived from C-terminal domain.

The secondary cell wall, which is mainly composed of cellulose, hemicellulose, and lignin, constitutes woody tissues and gives physical strength and hydrophobic properties for resistance against environmental stresses. We cloned and functionally analyzed the homologous transcription factor (TF) genes of SECONDARY WALL NAC (SWN) proteins from Hachiku bamboo (Phyllostachys nigra; PnSWNs). An RT-PCR analysis showed that PnSWNs are expressed in young tissues in bamboo. Their transcriptional activation activities were higher than that of the Arabidopsis NAC SECONDARY WALL THICKENING PROMOTING FACTOR 3 (NST3) TF, which was equivalent to SWN TFs in monocot. PnSWNs preferred to activate the genes related to secondary cell wall formation but not the genes related to programmed cell death. When PnSWNs were expressed in Arabidopsis, they highly induced secondary cell wall formation, like previously-shown rice SWN1. Dissection analysis revealed that this high activity largely depends on C-terminal domain. These results demonstrate that the cloned bamboo SWNs function as regulators of secondary cell wall formation with strong activation ability derived from C-terminal domain, and could be served as new genetic tools for secondary cell wall manipulation.

ETHYLENE RESPONSE FACTOR 34 promotes secondary cell wall thickening and strength of rice peduncles.

Cellulose and lignin are critical cell wall components for plant morphogenesis and adaptation to environmental conditions. The cytoskeleton supports cell wall deposition, but much of the underpinning regulatory components remain unknown. Here, we show that an APETALA2/ETHYLENE RESPONSE FACTOR (ERF) family transcription factor, OsERF34, directly promotes the expression of the actin- and microtubule-binding protein Rice Morphology Determinant (RMD) in rice (Oryza sativa) peduncles. OsERF34 and RMD are highly expressed in sclerenchymatous peduncle cells that are fortified by thick secondary cell walls (SCWs) that provide mechanical peduncle strength. erf34 and rmd-1 mutants contained lower cellulose and lignin contents and thinner SCWs, while ERF34 over-expressing (OE) lines maintained high cellulose and lignin content with thicker SCWs. These characteristics impacted peduncle mechanical strength, that is, reduced strength in erf34 and rmd-1 and increased strength of ERF34 OE plants. Taken together, our results demonstrate that the OsERF34-RMD cascade positively regulates SCW synthesis and mechanical strength in rice peduncles, which is important for yield, and provide a potential guide for improved peduncle breeding efforts in rice.

Identification and Expression Analysis of NAC Gene Family in Weeping Trait of Lagerstroemia indica

Lagerstroemia indica is a widely used ornamental plant in summer gardens because of its desirable plant shape. The weeping traits of plants are related to secondary cell wall thickness and hormone signaling. NAC (NAM-ATAF1/2-CUC2), as one of the plant-specific transcription factors, is a switch for the secondary cell wall and also involved in leaf senescence, phytohormone signaling, and other growth processes. We identified a total of 21 LiNAC genes from the transcriptome data, which we divided into 14 subgroups and 2 groups. The physicochemical characteristics of amino acids, subcellular localization, transmembrane structure, GO and KEGG enrichment, and expression patterns were also examined. The qRT-PCR analysis showed that the expressions of LiNAC8 and LiNAC13 in upright L. indica ‘Shaoguifei’ and weeping L. indica ‘Xiariwuniang’ were significantly higher from the beginning to the end of growth stage (S1–S3), and the expressions of ‘Shaoguifei’ were always higher than those of ‘Xiariwuniang’. However, LiNAC2 showed a downward trend in S1–S3 and the relative expression level of ‘Shaoguifei’ was lower than that of ‘Xiariwuniang’. It is hypothesized that these LiNAC genes may be involved in the regulation of weeping traits in L. indica. The results of this study provide a basis for analyzing the functions of LiNAC genes and help to explore the molecular regulatory mechanisms related to the weeping traits in L. indica.

The walnut shell network: 3D visualisation of symplastic and apoplastic transport routes in sclerenchyma tissue

Main conclusionHigh symplastic connectivity via pits was linked to the lignification of the developing walnut shell. With maturation, this network lessened, whereas apoplastic intercellular space remained and became relevant for shell drying.The shell of the walnut (Juglans regia) sclerifies within several weeks. This fast secondary cell wall thickening and lignification of the shell tissue might need metabolites from the supporting husk tissue. To reveal the transport capacity of the walnut shell tissue and its connection to the husk, we visualised the symplastic and apoplastic transport routes during shell development by serial block face-SEM and 3D reconstruction. We found an extensive network of pit channels connecting the cells within the shell tissue, but even more towards the husk tissue. Each pit channel ended in a pit field, which was occupied by multiple plasmodesmata passing through the middle lamella. During shell development, secondary cell wall formation progressed towards the interior of the cell, leaving active pit channels open. In contrast, pit channels, which had no plasmodesmata connection to a neighbouring cell, got filled by cellulose layers from the inner cell wall lamellae. A comparison with other nut species showed that an extended network during sclerification seemed to be linked to high cell wall lignification and that the connectivity between cells got reduced with maturation. In contrast, intercellular spaces between cells remained unchanged during the entire sclerification process, allowing air and water to flow through the walnut shell tissue when mature. The connectivity between inner tissue and environment was essential during shell drying in the last month of nut development to avoid mould formation. The findings highlight how connectivity and transport work in developing walnut shell tissue and how finally in the mature state these structures influence shell mechanics, permeability, conservation and germination.

A protoxylem pathway to evolution of pith? An hypothesis based on the Early Devonian euphyllophyte Leptocentroxyla.

The Early Devonian (Emsian, 400-395 Ma) tracheophyte Leptocentroxyla tetrarcha Bickner et Tomescu emend. Tomescu et McQueen combines plesiomorphic Psilophyton-type tracheid thickenings with xylem architecture intermediate between the plesiomorphic basal euphyllophyte haplosteles and the complex actinosteles of Middle Devonian euphyllophytes. We document xylem development in Leptocentroxyla based on anatomy and explore its implications, which may provide a window into the evolution of pith. Leptocentroxyla is preserved by permineralization in the Battery Point Formation (Quebec, Canada). Serial sections obtained using the cellulose acetate peel technique document branching pattern, anatomy of trace divergence to appendages, protoxylem architecture, and variations in tracheid size and wall thickening patterns. Leptocentroxyla has opposite decussate pseudo-whorled branching and mesarch protoxylem, and represents the earliest instance of central histological differentiation in a euphyllophyte actinostele. Tracheids at the centre of xylem exhibit simplified Psilophyton-type wall thickenings and are similar in size (at the axis centre) or smaller than the surrounding metaxylem tracheids (at the centre of appendage traces). The position and developmental attributes of the simplified Psilophyton-type tracheids suggest they may have been generated by the protoxylem developmental pathway. This supports the delayed and shortened protoxylem differentiation hypothesis, which explains the evolution of pith by (1) delay in the onset of differentiation and lengthening of cell growth duration in a central protoxylem strand; and (2) shortening of the interval of differentiation of those tracheids, leading to progressive simplification (and eventual loss) of secondary wall thickenings, and replacement of tracheids with a central parenchymatous area. NAC domain transcription factors and their interactions with abscisic acid may have provided the regulatory substrate for the developmental changes that led to the evolution of pith. These could have been orchestrated by selective pressures associated with the expansion of early vascular plants into water-stresses upland environments.

Study on histological structure and antimicrobial activity of Lilium candidum L

ABSTRACT Multidrug-resistant (MDR) bacterial infections have become a serious therapeutic problem recently, that can be solved by using of herbal drugs with antibacterial effects. The stem, leaf, tepal, anther, and ovary of Lilium candidum L. (white lily) were selected for histological and microbiological studies. For the microbial study, the stem, leaf, and tepal were extracted with methanol, n-hexane, chloroform, ethyl acetate, butanol, and water, and minimal inhibitory concentrations (MIC) were determined against standard and clinical strains (Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Salmonella Typhimurium, Pseudomonas aeruginosa, and Acinetobacter baumannii) by microdilution method after evaporation of the solvents. Unicellular non-glandular hairs were observed on the stem surface, and mesomorphic stomata were present in the stem and the leaf. The mesophyll of the leaf and the tepal is homogenous. Uneven secondary cell wall thickening was found in the endothecium of the anther. Placentation of the 3-locular ovary was axile. The butanol and water extracts of the stem showed an inhibitory activity on P. aeruginosa, while the butanol extract of the leaf inhibited the growth of S. aureus and E. coli, and its aqueous fraction inhibited S. aureus. The tepal extracts made with butanol and water showed an inhibitory effect on each tested strain. The observed antimicrobial activity of white lily yielded promising results, suggesting that further microbiological and phytochemical analyses are needed to explore the potential therapeutic use of the species.

Comparative Transcriptome Analysis Reveals Candidate Genes and Pathways for Potential Branch Growth in Elm (Ulmus pumila) Cultivars.

Simple SummaryElm (Ulmus pumila) is a strong essential wood, and it is widely used in cabinets, sculptures, and ship making. In the present study, phenotypic and comparative transcriptomic analyses were performed in elm fast- (UGu17 and UZuantian) and slow-growing cultivars (U81-07 and U82-39). Phenotypic observation showed that the thickness of secondary xylem of 2-year-old fast-growing branches was greater compared with slow-growing cultivars. Comparative transcriptome analysis predicted that many pathways were involved in vascular development and transcriptional regulation in elm, such as “plant-type secondary cell wall biogenesis”, “cell wall thickening”, and “phenylpropanoid biosynthesis”. NAC domain transcriptional factors (TFs) and their master regulators, cellulose synthase catalytic subunits (CESAs), xylan synthesis, and secondary wall thickness were presumed to function in the thickening mechanism of elm branches. Our results indicated that the general phenylpropanoid pathway and lignin metabolism had vital functions in the growth of elm branches.Wood plays a vital role in human life. It is important to study the thickening mechanism of tree branches and explore the mechanism of wood formation. Elm (Ulmus pumila) is a strong essential wood, and it is widely used in cabinets, sculptures, and ship making. In the present study, phenotypic and comparative transcriptomic analyses were performed in U. pumila fast- (UGu17 and UZuantian) and slow-growing cultivars (U81-07 and U82-39). Phenotypic observation showed that the thickness of secondary xylem of 2-year-old fast-growing branches was greater compared with slow-growing cultivars. A total of 9367 (up = 4363, down = 5004), 7159 (3413/3746), 7436 (3566/3870), and 5707 (2719/2988) differentially expressed genes (DEGs) were identified between fast- and slow-growing cultivars. Moreover, GO and KEGG enrichment analyses predicted that many pathways were involved in vascular development and transcriptional regulation in elm, such as “plant-type secondary cell wall biogenesis”, “cell wall thickening”, and “phenylpropanoid biosynthesis”. NAC domain transcriptional factors (TFs) and their master regulators (VND1/MYB26), cellulose synthase catalytic subunits (CESAs) (such as IRX5/IRX3/IRX1), xylan synthesis, and secondary wall thickness (such as IRX9/IRX10/IRX8) were supposed to function in the thickening mechanism of elm branches. Our results indicated that the general phenylpropanoid pathway (such as PAL/C4H/4CL) and lignin metabolism (such as HCL/CSE/CCoAOMT/CCR/F5H) had vital functions in the growth of elm branches. Our transcriptome data were consistent with molecular results for branch thickening in elm cultivars.

Anatomy of the hinge region of the petiole of prostrate leaves of the wintergreen fern Polystichum acrostichoides (Dryopteridaceae)

The reorientation of leaves of Polystichum acrostichoides (Michx.) Schott to a prostrate position is achieved by forming a hinge region near the base of the petiole. Anatomical details of the distal transition zone (DTZ), folding zone (FZ), and proximal transition zone (PTZ) of the hinge region were studied using light microscopy. Data showed that the vascular bundles (VBs) are intact throughout the hinge region. Towards the FZ, the DTZ and PTZ have an increasing number of cortical cells that underwent deformation and protoplast degeneration, and cell nuclei are apparent in strongly deformed cortical cells. In the FZ, the petiole is compressed, having a network of cell walls of dead cortical cells with patches of turgid cortical cells, and largely intact endodermis surrounding the VBs. The helical secondary wall thickenings of the protoxylem and scalariform thickenings of the metaxylem may contribute to the flexibility of the VBs. This study provided evidence for the deformation and degeneration of cortical cells, which is responsible for the loss of turgidity of the hinge region while their cell walls may provide cushioning during bending and insulation for the living tissues under the snowpack, illustrating a unique strategy of ferns to adapt to a cold environment.

Melatonin enhances stem strength by increasing lignin content and secondary cell wall thickness in herbaceous peony.

Cut flower quality is severely restrained by stem bending due to low stem strength. Melatonin has been shown to function in many aspects of plant growth and development, yet whether it can enhance stem strength, and the corresponding underlying mechanisms remain unclear. We investigated the role of melatonin in enhancement of stem strength in herbaceous peony (Paeonia lactiflora Pall.) by applying exogenous melatonin and changing endogenous melatonin biosynthesis. Endogenous melatonin content positively correlated with lignin content and stem strength in various P. lactiflora cultivars. Supplementation with exogenous melatonin significantly enhanced stem strength by increasing lignin content and the S/G lignin compositional ratio, up-regulating lignin biosynthetic gene expression. Moreover, overexpression of TRYPTOPHAN DECARBOXYLASE GENE (TDC) responsible for the first committed step of melatonin biosynthesis in tobacco, significantly increased endogenous melatonin, which further increased the S/G ratio and stem strength. In contrast, silencing PlTDC in P. lactiflora decreased endogenous melatonin, the S/G ratio and stem strength. Finally, manipulating the expression of CAFFEIC ACID O-METHYLTRANSFERASE GENE (COMT1), which is involved in both melatonin and lignin biosynthesis, showed even greater effects on melatonin, the S/G ratio and stem strength. Our results suggest that melatonin has a positive regulatory effect on P. lactiflora stem strength.

Analysis of WRKY Resistance Gene Family in Boehmeria nivea (L.) Gaudich: Crosstalk Mechanisms of Secondary Cell Wall Thickening and Cadmium Stress.

A total of 60 WRKY family genes of ramie were identified in the ramie. The genes were unevenly distributed across 14 chromosomes in the specie and highly concentrated (72%) in the distal telomeric region. Phylogenetic analysis placed these genes into seven distinct subfamilies groups: I, II (a, b, c, d, e), and III, with group IIc containing only the variant of heptapetide sequence (WRKYGKK). Segmental duplication events (41.7%) was found to be the main driver of BnGWRKY evolution. Thirty eight from among the genes showed collinear relationships with WRKY genes from Arabidopsis thaliana, Cannabis sativa, Oryza sativa, and Zea mays. The number and density of stress and hormone responsives cis-acting elements were comparably higher than other elements, with abundant ARE and rare LTR cis-acting elements indicating the long-standing adaptability of ramie to its natural environment. GO and KEGG enrichment analysis of the WRKY target genes revealed their involvement in response to stimuli, immune system processes, transporter protein activity and antioxidant activity. Expression analysis show that most WRKYs were activated by the cadmium stress, more especially the BnGWRKY2, BnGWRKY15, BnGWRKY20, BnGWRKY50 and BnGWRKY58. Combining transcriptome, orthologous gene relationships and qPCR result, we established the possible involvement of BnGWRKY50 and BnGWRKY58 in crosstalk mechanism between secondary cell wall thickening and Cd2+ stress. This provided information into the role of BnGWRKY proteins in ramie secondary wall development and cadmium stress response to, and could serve as basis for improvement of the ramie.

Comparative transcriptome analysis reveals the regulation network for fiber strength in cotton.

Determine the effect of secondary cell wall (SCW) thickness and microcrystalline cellulose content (MCC) on mature fiber strength (FS) and reveal through comparative transcriptome analysis the molecular regulation network governing FS in cotton. Transmission electron microscope (TEM) analysis of two parent varieties, Prema with elite FS and 86-1 with weak fiber, revealed significant difference in the SCW but not in MCC. Transcriptome analysis revealed that genes differentially expressed during SCW thickening (20 DPA) are highly related to FS; in particular, up-regulated genes such as UDPG, CESA2, and NAC83 were important in SCW thickening, likely contributing to higher FS. GO and KEGG enrichment analysis revealed the common up-regulated genes to be enriched in carbon metabolism and terms relating to the cell wall. We developed two recombinant inbred lines with elite FS, selected from the filial generation of Prema and 86-1. By comparing transcriptomic data, we revealed the gene expression network governing SCW thickness in mature fiber. Our results provide solid insights into the relationship of the SCW and FS.

Ectopic expression of Populus MYB10 promotes secondary cell wall thickening and inhibits anthocyanin accumulation

Secondary cell wall (SCW) formation is regulated by a multilevel transcriptional regulatory network, in which MYB transcription factors (TFs) play key roles. In woody plants, hundreds of MYB TFs have been identified, most of which have unknown functions in wood SCW biosynthesis. Here, we characterized the function of a Populus MYB gene, PtoMYB10. PtoMYB10 was found to encode an R2R3-MYB TF and exhibit dominant expression in xylem tissues. PtoMYB10 was determined to be located in the nucleus with the ability to activate transcription. Overexpression of PtoMYB10 in Populus resulted in a drastic increase in SCW thickening in xylem fiber cells as well as ectopic deposition of lignin in cortex cells. The expression of genes associated with lignin biosynthesis was induced in PtoMYB10 overexpressing plants, whereas repressed gene expression was found with the anthocyanin biosynthesis pathway. Lignin and anthocyanin are both produced from metabolites of the phenylpropanoid pathway. Accordingly, the anthocyanin content of Populus overexpressing PtoMYB10 decreased by more than 68%. These results indicate that PtoMYB10 can positively regulate xylary fiber SCW thickening, accompanied by the reprogramming of phenylpropanoid metabolism, which redirects metabolic flux from anthocyanin biosynthesis to monolignol biosynthesis.

A natural mutation of the NST1 gene arrests secondary cell wall biosynthesis in the seed coat of a hull-less pumpkin accession.

Hull-less pumpkins (Cucurbita pepo L.) are naturally occurring novel variants known as oilseed or naked-seeded pumpkins, and are characterized by the absence of a normal lignified seed coat. Due to a specialized seed coat structure, these variants serve as a good model for studying seed coat formation and simplify the processing of pumpkin seeds. However, causal genes for this hull-less trait still remain unknown. Here, by bulked segregant analysis and fine mapping, we found that mutation of a single gene, NAC SECONDARY WALL THICKENING PROMOTING FACTOR 1 (NST1), accounts for the hull-less trait. A 14-bp sequence insertion in the CpNST1 gene causes premature termination of CpNST1 translation, leading to lack of secondary cell wall (SCW) biosynthesis in hull-less seed coats. In situ hybridization analysis provided further evidence for the role of CpNST1 in pumpkin seed coat SCW biosynthesis. Interestingly, through secondary cell wall compositional analysis, we found that the main SCW components differed among cell layers in the seed coat. RNA-seq analysis indicated an upstream role of CpNST1 in the SCW biosynthesis network. Collectively, our findings provide mechanistic insight into seed coat SCW biosynthesis, and a target gene for breeders to introduce this hull-less trait for commercial exploitation.

The role of senescence-associated gene101 (PagSAG101a) in the regulation of secondary xylem formation in poplar.

Wood is produced by the accumulation of secondary xylem via proliferation and differentiation of the cambium cells in woody plants. Identifying the regulators involved in this process remains a challenging task. In this study, we isolated PagSAG101a, the homolog of Arabidopsis thaliana SAG101, from a hybrid poplar (Populus alba × Populus glandulosa) clone 84K and investigated its role in secondary xylem development. PagSAG101a was expressed predominantly in lignified stems and localized in the nucleus. Compared with non-transgenic 84K plants, transgenic plants overexpressing PagSAG101a displayed increased plant height, internode number, stem diameter, xylem width, and secondary cell wall thickness, while opposite phenotypes were observed for PagSAG101a knock-out plants. Transcriptome analyses revealed that differentially expressed genes were enriched for those controlling cambium cell division activity and subsequent secondary cell wall deposition during xylem formation. In addition, the tandem CCCH zinc finger protein PagC3H17, which positively regulates secondary xylem width and secondary wall thickening in poplar, could bind to the promoter of PagSAG101a and mediate the regulation of xylem differentiation. Our results support that PagSAG101a, downstream of PagC3H17, functions in wood development.