Effects of uniconazole with or without micronutrient on the lignin biosynthesis,lodging resistance,and winter wheat production in semiarid regions

Lodging stress results in grain yield and quality reduction in wheat. Uniconazole, a potential plant growth regulator significantly enhances lignin biosynthesis and thus provides mechanical strength to plants in order to cope with lodging stress. A field study was conducted during the 2015–2016 and 2016–2017 growing seasons, to investigate the effects of uniconazole sole application or with micronutrient on the lignin biosynthesis, lodging resistance, and production of winter wheat. In the first experiment, uniconazole at concentrations of 0(CK), 15(US1), 30(US2), and 45(US3) mg L^-1 was applied as sole seed soaking, while in the second experiment with manganese(Mn) at concentration of 0.06 g L^-1 Mn, 0.06 g L^-1 Mn+ 15 mg L^-1 uniconazole(UMS1), 0.06 g L^-1 Mn+30 mg L^-1 uniconazole(UMS2), and 0.06 g L^-1 Mn+45 mg L^-1 uniconazole(UMS3), respectively. Uniconazole sole application or with micronutrient significantly increased the lignin content by improving the lignin-related enzyme activities of phenylalanine ammonia-lyase, cinnamyl alcohol dehydrogenase, tyrosine ammonialyase, and peroxidase, ameliorating basal internode characteristics, and breaking strength. The spike length, spike diameter, spikes/plant, weight/spike, yield/spike, and grain yield increased and then decreased with uniconazole application at a higher concentration, where their maximum values were recorded with UMS2 and US2 treatments. The lignin accumulation was positively correlated with lignin-related enzyme activities and breaking strength while, negatively correlated with lodging rate. Uniconazole significantly improved the lignin biosynthesis, lodging resistance, and grain yield of winter wheat and the treatments which showed the greatest effects were uniconazole seed soaking with micronutrient at a concentration of 30 mg L^-1 and 0.06 g L^-1 , and uniconazole sole seed soaking at a concentration of 30 mg L^-1 .

Effect of shade stress on lignin biosynthesis in soybean stems

To clarify how shade stress affects lignin biosynthesis in soybean stem, two varieties, Nandou 12（shade tolerant） and Nan 032-4（shade susceptible） grew under normal light and shade conditions（the photosynthetically active radiation and the ratio of red：far-red were lower than normal light condition）. Lignin accumulation, transcripts of genes involved in lignin biosynthesis, and intermediates content of lignin biosynthesis were analyzed. Both soybean varieties suffered shade stress had increased plant heights and internode lengths, and reduced stem diameters and lignin accumulation in stems. The expression levels of lignin-related genes were significantly influenced by shade stress, with interactions between the light environment and variety. The gene of 3-hydroxylase（C3H）, cinnamoyl-Co A reductase（CCR）, caffeoylCoAO-methyltransferase（CCoAOMT）, and peroxidase（POD） attributed to lignin biosynthesis under shade stress, and the down-regulation of these genes resulted in lower caffeic, sinapic, and ferulic acid levels, which caused a further decrease in lignin biosynthesis. Under shade stress, the shade tolerant soybean variety（Nandou 12） showed stiffer stems, higher lignin content, and greater gene expression level and higher metabolite contents than shade susceptible one. So these characteristics could be used for screening the shade-tolerant soybean for intercropping.

Upgraded durian genome reveals the role of chromosome reshuffling during ancestral karyotype evolution,lignin biosynthesis regulation,and stress tolerance

Durian(Durio zibethinus)is a tropical fruit that has a unique flavor and aroma.It occupies a significant phylogenetic position within the Malvaceae family.Extant core-eudicot plants are reported to share seven ancestral karyotypes that have undergone reshuffling,resulting in an abundant genomic diversity.However,the ancestral karyotypes of the Malvaceae family,as well as the evolution trajectory leading to the28 chromosomes in durian,remain poorly understood.Here,we report the high-quality assembly of the durian genome with comprehensive comparative genomic analyses.By analyzing the collinear blocks between cacao and durian,we inferred 11 Malvaceae ancestral karyotypes.These blocks were present in a single-copy form in cacao and mainly in triplicates in durian,possibly resulting from a recent whole genome triplication(WGT)event that led to hexaploidization of the durian genome around 20(17–24)million years ago.A large proportion of the duplicated genes in durian,such as those involved in the lignin biosynthesis module for phenylpropane biosynthesis,are derived directly from whole genome duplication,which makes it an important force in reshaping its genomic architecture.Transcriptome studies have revealed that genes involved in feruloyl-Co A formations were highly preferentially expressed in fruit peels,indicating that the thorns produced on durian fruit may comprise guaiacyl and syringyl lignins.Among all the analyzed transcription factors(TFs),members of the heat shock factor family(HSF)were the most significantly upregulated under heat stress.All subfamilies of genes encoding heat shock proteins(HSPs)in the durian genome appear to have undergone expansion.The potential interactions between HSF Dzi05.397 and HSPs were examined and experimentally verified.Our study provides a high-quality durian genome and reveals the reshuffling mechanism of ancestral Malvaceae chromosomes to produce the durian genome.We also provide insights into the mechanism underlying lignin biosynthesis and heat stress tolerance.

PagERF81 regulates lignin biosynthesis and xylem cell differentiation in poplar

Lignin is a major component of plant cell walls and is essential for plant growth and development. Lignin biosynthesis is controlled by a hierarchical regulatory network involving multiple transcription factors. In this study, we showed that the gene encoding an APETALA 2/ethylene-responsive element binding factor(AP2/ERF) transcription factor, PagERF81,from poplar 84 K(Populus alba × P. glandulosa) is highly expressed in expanding secondary xylem cells. Two independent homozygous Pagerf81 mutant lines created by gene editing, produced significantly more but smaller vessel cells and longer fiber cells with more lignin in cell walls, while PagERF81 overexpression lines had less lignin,compared to non-transgenic controls. Transcriptome and reverse transcription quantitative PCR data revealed that multiple lignin biosynthesis genes including Cinnamoyl CoA reductase 1(PagCCR1),Cinnamyl alcohol dehydrogenase 6(PagCAD6), and 4-Coumarate-CoA ligase-like 9(Pag4CLL9) were upregulated in Pagerf81 mutants, but down-regulated in PagERF81 overexpression lines. In addition, a transient transactivation assay revealed that PagERF81 repressed the transcription of these three genes.Furthermore, yeast one hybrid and electrophoretic mobility shift assays showed that PagERF81 directly bound to a GCC sequence in the PagCCR1 promoter. No known vessel or fiber cell differentiation related genes were differentially expressed, so the smaller vessel cells and longer fiber cells observed in the Pagerf81 lines might be caused by abnormal lignin deposition in the secondary cell walls. This study provides insight into the regulation of lignin biosynthesis, and a molecular tool to engineer wood with high lignin content, which would contribute to the lignin-related chemical industry and carbon sequestration.

Lignin Biosynthesis Studies in Plant Tissue Cultures

Lignin, a phenolic polymer abundant in cell walls of certain cell types, has given challenges to scientists studying its structure or biosynthesis. In plants lignified tissues are distributed between other, non-lignified tissues, Characterization of native lignin in the cell wall has been difficult due to the highly cross-linked nature of the wall components. Model systems, like plant tissue cultures with tracheary element differentiation or extracellular lignin formation, have provided useful information related to lignin structure and several aspects of lignin formation. For example, many enzyme activities in the phenylpropanoid pathway have been first identified in tissue cultures. This review focuses on studies where the use of plant tissue cultures has been advantageous in structural and biosynthesis studies of lignin, and discusses the validity of tissue cultures as models for lignin biosynthesis.

Overexpression of the peroxidase gene ZmPRX1 increases maize seedling drought tolerance by promoting root development and lignification

Drought is a main abiotic stress factor hindering plant growth,development,and crop productivity.Therefore,it is crucial to understand the mechanisms by which plants cope with drought stress.Here,the function of the maize peroxidase gene ZmPRX1 in drought stress tolerance was investigated by measurement of its expression in response to drought treatment both in a ZmPRX1 overexpression line and a mutant line.The higher root lignin accumulation and seedling survival rate of the overexpression line than that of the wild type or mutant support a role for ZmPRX1 in maize drought tolerance by regulating root development and lignification.Additionally,yeast one-hybrid,Dule luciferase and ChIP-qPCR assays showed that ZmPRX1 is negatively regulated by a nuclear-localized ZmWRKY86 transcription factor.The gene could potentially be used for breeding of drought-tolerant cultivars.

Effect of the suppression of BpAP1 on the expression of lignin related genes in birch

Lignin is an integral part of secondary cell walls in plants and plays important roles in maintaining the strength of stems,enhancing transport ability of stems,and providing resistance to multiple stresses.Lignin biosynthesis has become one of the hotspots in molecular forest biology research.The AP1 transcription factor plays important roles in plant fower development.However,in this study,suppression of BpAP1 altered the transcription profiles of white birch and RNA-seq was used to find that suppression of BpAP1 changed the expression of lignin pathway-related genes;C4 H/CYP73A,POD were down-regulated and HCT,CCoAOMT,REF1 and CAD were up-regulated.Cell walls of the suppressed transgenic birch were significantly thinner than the wild type of birch,and Bp AP1-repressed birch contained less lignin.In addition to regulation of foral development,BpAP1 might play a role in regulating the expression of genes in lignin biosynthesis of birch.This study could provide a new insight into the function of AP1 genes in woody species.

The application of Fourier transform mid-infrared(FTIR) spectroscopy to identify variation in cell wall composition of Setaria italica ecotypes

Cell wall composition in monocotyledonous grasses has been identified as a key area of research for developing better feedstocks for forage and biofuel production.Setaria viridis and its close domesticated relative Setaria italica have been chosen as suitable monocotyledonous models for plants possessing the C4 pathway of photosynthesis including sorghum,maize,sugarcane,switchgrass and Miscanthus×giganteus.Accurate partial least squares regression（PLSR）models to predict S.italica stem composition have been generated,based upon Fourier transform mid-infrared（FTIR）spectra and calibrated with wet chemistry determinations of ground S.italica stem material measured using a modified version of the US National Renewable Energy Laboratory（NREL）acid hydrolysis protocol.The models facilitated a high-throughput screening analysis for glucan,xylan,Klason lignin and acid soluble lignin（ASL）in a collection of 183 natural S.italica variants and clustered them into classes,some possessing unique chemotypes.The predictive models provide a highly efficient screening tool for large scale breeding programs aimed at identifying lines or mutants possessing unique cell wall chemotypes.Genes encoding key catalytic enzymes of the lignin biosynthesis pathway exhibit a high level of conservation with matching expression profiles,measured by RT-q PCR,among accessions of S.italica,which closely mirror profiles observed in the different developmental regions of an elongating internode of S.viridis by RNASeq.

Simultaneously disrupting AtPrx2,AtPrx25 and AtPrx71 alters lignin content and structure in Arabidopsis stem

Plant class III heme peroxidases catalyze lignin polymerization. Previous reports have shown that at least three Arabidopsis thaliana peroxidases, AtPrx2, AtPrx25 and AtPrx71, are involved in stem lignification using T-D NA insertion mutants, atprx2, atprx25, and atprx71. Here, we generated three double mutants, atprx2/atprx25, atprx2/atprx71, and atprx25/atprx71, and investigated the impact of the simultaneous deficiency of these peroxidases on lignins and plant growth. Stem tissue analysis using the acetyl bromide method and derivatization followed by reductive cleavage revealed improved lignin characteristics, such as lowered lignin content and increased arylglycerol-β-aryl （β-O-4） linkage type, especially β-O-4 linked syringyl units, in lignin, supporting the roles of these genes in lignin polymerization. In addition, none of the double mutants oexhibited severe growth defects, such as shorter plant stature, dwarfing, or sterility, and their stems had improved ceil wall degradability. This study will contribute to progress in lignin bioengineering to improve iignocellulosic biomass.

Functional Genomics of Wood Quality and Properties

Genomics promises to enrich the investigations of biology and biochemistry. Current advancements in genomics have major implications for genetic improvement in animals, plants, and microorganisms, and for our understanding of cell growth, development, differentiation, and communication. Significant progress has been made in the understanding of plant genomics in recent years, and the area continues to progress rapidly. Functional genomics offers enormous potential to tree improvement and the understanding of gene expression in this area of science worldwide. In this review we focus on functional genomics of wood quality and properties in trees, mainly based on progresses made in genomics study of Pinus and Populus. The aims of this review are to summarize the current status of functional genomics including: (1) Gene discovery; (2) EST and genomic sequencing; (3) From EST to functional genomics; (4) Approaches to functional analysis; (5) Engineering lignin biosynthesis; (6) Modification of cell wall biogenesis; and (7) Molecular modelling. Functional genomics has been greatly invested worldwide and will be important in identifying candidate genes whose function is critical to all aspects of plant growth, development, differentiation, and defense. Forest biotechnology industry will significantly benefit from the advent of functional genomics of wood quality and properties.