Wood Property Traits Research Articles

Identification of superior hybrid clones for fibre biometry in Eucalyptus camaldulensis × E. tereticornis using multi trait stability index

Abstract Genotype × environment (G×E) interaction is a major challenge in selecting superior genotypes based on growth traits in Eucalyptus since phenotypic variability is significantly affected by environmental heterogeneity. The aim of the present investigation was to understand the relationship between wood property traits and fibre biometry in the bi-parental mapping population of E. camaldulensis × E. tereticornis across three locations and identify stable genotypes based on multiple traits to improve prediction accuracy in breeding programs. High broad-sense heritability was documented for fibre parameters indicating a good prospect of these traits for genotype selection in hybrid breeding programmes in Eucalyptus. Significant positive correlation of fibre parameters with cellulose, acoustic velocity, DMoE and negative correlation with lignin reiterate that the fibre traits can be improved by the selection of genotype for improved wood property traits. Multi-Trait Stability Index (MTSI) and weighted average of absolute scores of the genotype index (WAASB) short-listed four genotypes (C343, C327, C246 and C161) with improved wood property traits and the mean of selected genotypes for all traits was significantly higher than the grand mean of the overall genotypes. The identified superior and stable genotypes with improved wood properties and fibre biometry can be used in plantation programs or as genitors in breeding programs.

Selection for Both Growth and Wood Properties in Chinese Fir Breeding Parents Based on a 6-Year Grafted Clone Test

With the growing demand for high-quality timber, selection processes for both growth and wood properties are needed for multi-trait breeding programs in Chinese fir (Cunninghamia lanceolata (Lamb.) Hook.). The present study examined the variation and correlation of growth (tree height, diameter at breast height, stem volume, crown-width) and wood properties (wood basic density, hygroscopicity, and heart-wood ratio) traits of 201 Chinese fir breeding parents, aiming to select better parents for future multi-trait improvement. The results showed that significant differences (p < 0.01) regarding growth and wood property traits were observed among clones in an individual site and in a two-site joint analysis. The repeatability of the tested traits varied from 0.22 to 0.87. Strong and positive (p < 0.01) correlations were detected among the four growth traits, while wood basic density had a significant negative correlation (p < 0.01 or 0.05) with the growth traits. A set of parent clones was shortlisted with substantial realized gains (ranging from 4.59% to 83.77%) in growth and wood traits. It was suggested that these selected parents could be used to improve the growth and wood quality of Chinese fir.

High-density genetic linkage mapping reveals low stability of QTLs across environments for economic traits in Eucalyptus.

Eucalyptus urophylla, E. tereticornis and their hybrids are the most important commercial forest tree species in South China where they are grown for pulpwood and solid wood production. Construction of a fine-scale genetic linkage map and detecting quantitative trait loci (QTL) for economically important traits linked to these end-uses will facilitate identification of the main candidate genes and elucidate the regulatory mechanisms. A high-density consensus map (a total of 2754 SNPs with 1359.18 cM) was constructed using genotyping by sequencing (GBS) on clonal progenies of E. urophylla × tereticornis hybrids. QTL mapping of growth and wood property traits were conducted in three common garden experiments, resulting in a total of 108 QTLs. A total of 1052 candidate genes were screened by the efficient combination of QTL mapping and transcriptome analysis. Only ten QTLs were found to be stable across two environments, and only one (qSG10Stable mapped on chromosome 10, and associated with lignin syringyl-to-guaiacyl ratio) was stable across all three environments. Compared to other QTLs, qSG10Stable explained a very high level of phenotypic variation (18.4-23.6%), perhaps suggesting that QTLs with strong effects may be more stably inherited across multiple environments. Screened candidate genes were associated with some transcription factor families, such as TALE, which play an important role in the secondary growth of plant cell walls and the regulation of wood formation. While QTLs such as qSG10Stable, found to be stable across three sites, appear to be comparatively uncommon, their identification is likely to be a key to practical QTL-based breeding. Further research involving clonally-replicated populations, deployed across multiple target planting sites, will be required to further elucidate QTL-by-environment interactions.

Genetic architecture behind developmental and seasonal control of tree growth and wood properties in Norway spruce.

Genetic control of tree growth and wood formation varies depending on the age of the tree and the time of the year. Single-locus, multi-locus, and multi-trait genome-wide association studies (GWAS) were conducted on 34 growth and wood property traits in 1,303 Norway spruce individuals using exome capture to cover ~130K single-nucleotide polymorphisms (SNPs). GWAS identified associations to the different wood traits in a total of 85 gene models, and several of these were validated in a progenitor population. A multi-locus GWAS model identified more SNPs associated with the studied traits than single-locus or multivariate models. Changes in tree age and annual season influenced the genetic architecture of growth and wood properties in unique ways, manifested by non-overlapping SNP loci. In addition to completely novel candidate genes, SNPs were located in genes previously associated with wood formation, such as cellulose synthases and a NAC transcription factor, but that have not been earlier linked to seasonal or age-dependent regulation of wood properties. Interestingly, SNPs associated with the width of the year rings were identified in homologs of Arabidopsis thaliana BARELY ANY MERISTEM 1 and rice BIG GRAIN 1, which have been previously shown to control cell division and biomass production. The results provide tools for future Norway spruce breeding and functional studies.

Integrating genome-wide association mapping of additive and dominance genetic effects to improve genomic prediction accuracy in Eucalyptus.

Genome-wide association studies (GWAS) is a powerful and widely used approach to decipher the genetic control of complex traits. Still, a significant challenge for dissecting quantitative traits in forest trees is statistical power. This study uses a population consisting of 1,123 samples derived from two successive generations of crosses between Eucalyptus grandis (W. Hill) and E. urophylla (S.T. Blake). All samples have been phenotyped for growth and wood property traits and genotyped using the EuChip60K chip, yielding 37,832 informative single nucleotide polymorphisms (SNPs). We use multi-locus GWAS models to assess additive and dominance effects to identify markers associated with growth and wood property traits in the eucalypt hybrids. Additive and dominance association models identified 78 and 82 significant SNPs across all traits, respectively, which captured between 39 and 86% of the genomic-based heritability. We also used SNPs identified from the GWAS and SNPs using less stringent significance thresholds to evaluate predictive abilities in a genomic selection framework. Genomic selection models based on the top 1% SNPs captured a substantially greater proportion of the genetic variance of traits compared with when we used all SNPs for model training. The prediction ability of estimated breeding values improved significantly for all traits when using either the top 1% SNPs or SNPs identified using a relaxed p value threshold (p<10-3 ). This study also highlights the added value of incorporating dominance effects for identifying genomic regions controlling growth traits in trees. Moreover, integrating GWAS results into genomic selection method provides enhanced power relative to discrete associations for identifying genomic variation potentially valuable for forest tree breeding.

Genetic variation analysis and comprehensive evaluation of wood property traits of 20-year-old Chinese fir clone

Genetic variation analysis and comprehensive evaluation of wood property traits of 20-year-old Chinese fir clone

Targeted re-sequencing and genome-wide association analysis for wood property traits in breeding population of Eucalyptus tereticornis × E. grandis

Globally, Eucalyptus plantations occupy 22 million ha area and is one of the preferred hardwood species due to their short rotation, rapid growth, adaptability and wood properties. In this study, we present results of GWAS in parents and 100 hybrids of Eucalyptus tereticornis × E. grandis using 762 genes presumably involved in wood formation. Comparative analysis between parents predicted 32,202 polymorphic SNPs with high average read depth of 269-562× per individual per nucleotide. Seventeen wood related traits were phenotyped across three diverse environments and GWAS was conducted using 13,610 SNPs. A total of 45 SNP-trait associations were predicted across two locations. Seven large effect markers were identified which explained more than 80% of phenotypic variation for fibre area. This study has provided an array of candidate genes which may govern fibre morphology in this genus and has predicted potential SNPs which can guide future breeding programs in tropical Eucalyptus.

Association Study and Mendelian Randomization Analysis Reveal Effects of the Genetic Interaction Between PtoMIR403b and PtoGT31B-1 on Wood Formation in Populus tomentosa.

MicroRNAs (miRNAs), important posttranscriptional regulators of gene expression, play a crucial role in plant growth and development. A single miRNA can regulate numerous target genes, making the determination of its function and interaction with targets challenging. We identified PtomiR403b target to PtoGT31B-1, which encodes a galactosyltransferase responsible for the biosynthesis of cell wall polysaccharides. We performed an association study and epistasis and Mendelian randomization (MR) analyses to explore how the genetic interaction between PtoMIR403b and its target PtoGT31B-1 underlies wood formation. Single nucleotide polymorphism (SNP)-based association studies identified 25 significant associations (P < 0.01, Q < 0.05), and PtoMIR403b and PtoGT31B-1 were associated with five traits, suggesting a role for PtomiR403b and PtoGT31B-1 in wood formation. Epistasis analysis identified 93 significant pairwise epistatic associations with 10 wood formation traits, and 37.89% of the SNP-SNP pairs indicated interactions between PtoMIR403b and PtoGT31B-1. We performed an MR analysis to demonstrate the causality of the relationships between SNPs in PtoMIR403b and wood property traits and that PtoMIR403b modulates wood formation by regulating expression of PtoGT31B-1. Therefore, our findings will facilitate dissection of the functions and interactions with miRNA-targets.

Population Divergence along a Genetic Line of Least Resistance in the Tree Species Eucalyptus globulus.

The evolutionary response to selection depends on the distribution of genetic variation in traits under selection within populations, as defined by the additive genetic variance-covariance matrix (G). The structure and evolutionary stability of G will thus influence the course of phenotypic evolution. However, there are few studies assessing the stability of G and its relationship with population divergence within foundation tree species. We compared the G-matrices of Mainland and Island population groups of the forest tree Eucalyptus globulus, and determined the extent to which population divergence aligned with within-population genetic (co)variation. Four key wood property traits exhibiting signals of divergent selection were studied—wood density, extractive content, and lignin content and composition. The comparison of G-matrices of the mainland and island populations indicated that the G-eigenstructure was relatively well preserved at an intra-specific level. Population divergence tended to occur along a major direction of genetic variation in G. The observed conservatism of G, the moderate evolutionary timescale, and close relationship between genetic architecture and population trajectories suggest that genetic constraints may have influenced the evolution and diversification of the E. globulus populations for the traits studied. However, alternative scenarios, including selection aligning genetic architecture and population divergence, are discussed.

Genome Cytosine Methylation May Affect Growth and Wood Property Traits in Populations of Populus tomentosa

Growth and wood formation are crucial and complex biological processes during tree development. These biological regulatory processes are presumed to be controlled by DNA methylation. However, there is little direct evidence to show that genes taking part in wood regulation are affected by cytosine methylation, resulting in phenotypic variations. Here, we detected epimarkers using a methylation-sensitive amplification polymorphism (MSAP) method and performed epimarker–trait association analysis on the basis of nine growth and wood property traits within populations of 432 genotypes of Populus tomentosa. Tree height was positively correlated with relative full-methylation level, and 1101 out of 2393 polymorphic epimarkers were associated with phenotypic traits, explaining 1.1–7.8% of the phenotypic variation. In total, 116 epimarkers were successfully sequenced, and 96 out of these sequences were linked to putative genes. Among them, 13 candidate genes were randomly selected for verification using quantitative real-time PCR (qRT-PCR), and it also showed the expression of nine putative genes of PtCYP450, PtCpn60, PtPME, PtSCP, PtGH, PtMYB, PtWRKY, PtSTP, and PtABC were negatively correlated with DNA methylation level. Therefore, it suggested that changes in DNA methylation might contribute to regulating tree growth and wood property traits.

Effects of cutting size on the growth and wood property traits of short-rotation coppice willows

Short-rotation coppice willows, which are commonly propagated with cuttings, are promising bioenergy resource crops. In this study, a field trial with a completely randomized block design was carried out with 787 clones from three full-sib pedigrees involving two shrub willow species. A total of 7083 cuttings were planted in the field, and the mass of each cutting was recorded correspondingly. The mortality rate, growth traits, and wood property traits of each cutting-propagated plant were measured throughout a complete growing season. The obtained data showed that heavier cuttings produced shoots with lower mortality rate, higher height, thicker ground diameter, and larger biomass production than lighter cuttings. The cutting mass significantly improved plant performance, especially at the early developmental stages. However, the cutting mass had no effect on the basic density and the main chemical composition (i.e., cellulose, hemicellulose, and lignin) of willow wood. This study provided essential information for improving the establishment of short-rotation coppice willow plantations.

Evolution and Functional Divergence of the Fructokinase Gene Family in Populus.

New kinase has emerged throughout evolution, but how new kinase evolve while maintaining their functions and acquiring new functions remains unclear. Fructokinase (FRK), the gateway kinase to fructose metabolism, plays essential roles in plant development, and stress tolerance. Here, we explored the evolution of FRK gene family in 20 plant species (from green algae to angiosperms) and their functional roles in Populus. We identified 125 putative FRK genes in the 20 plant species with an average of 6 members per species. Phylogenetic analysis separated these 125 genes into 8 clades including 3 conserved clades and 5 specific clades, the 5 of which only exist in green algae or angiosperms. Evolutionary analysis revealed that FRK genes in ancient land plants have the largest number of functional domains with the longest amino acid sequences, and the length of FRK genes became shorter during the transition to vascular plants. This was accompanied by loss, acquisition, and diversification of functional domains. In Populus, segmental duplication appears to be the main mechanism for the expansion of FRK genes. Specially, most FRK genes duplicated in salicoids are regulated by Populus-specific microRNAs. Furthermore, compared with common FRKs, Populus-specific FRKs have showed higher expression specificity and are associated with fewer growth and wood property traits, which suggests that these FRKs may have undergone functional divergence. Our study explores the specific roles of FRKs in the Populus genome and provides new insights for functional investigation of this gene family.

Linkage-linkage disequilibrium dissection of the epigenetic quantitative trait loci (epiQTLs) underlying growth and wood properties in Populus.

Increasing evidence indicates that DNA methylation is heritable and serves as an essential marker contributing to phenotypic variation. Linkage-linkage disequilibrium mapping was used to decipher the epigenetic architecture underlying nine growth and wood property traits in a linkage population (550 F1 progeny) and a natural population (435 unrelated individuals) of Populus using methylation-sensitive amplification polymorphism (MSAP)-based analysis. The interactions between genetic and epigenetic variants in the causative genes was further unveiled using expression quantitative trait methylation (eQTM) and nucleotide (eQTN) mapping strategies. A total of 163 epigenetic quantitative trait loci (epiQTLs; LOD≥3.0), explaining 1.7-44.5% of phenotypic variations, were mapped to a high-resolution epigenetic map with 19 linkage groups, which was supported by the significant MSAP associations (P<0.001) in the two populations. There were 23 causal genes involved in growth regulation and wood formation, whose markers were located in epiQTLs and associated with the same traits in both populations. Further eQTN and eQTM mapping showed that causal genetic and epigenetic variants within the 23 candidate genes may interact more in trans in gene expression and phenotype. The present study provides strategies for investigating epigenetic architecture and the interaction between genetic and epigenetic variants modulating complex traits in forest trees.

Mapping quantitative trait loci for growth and wood property traits in Cryptomeria japonica across multiple environments

Genomic regions which affected tree growth and wood property traits were investigated in the major plantation tree of Japan, Cryptomeria japonica, in three replicated common garden experiments planted in contrasting environments in Kyushu and Honshu, Japan. Phenotypic traits measured were stem diameter at breast height, tree height, wood strength (Young’s modulus), heartwood density, sapwood density, heartwood moisture content, and sapwood moisture content. Quantitative trait locus (QTL) analysis identified an average of 53 QTLs across the three environments. There were two QTLs which affected the same traits across all three environments. These stable QTLs were identified as being associated with sapwood density and Young’s modulus and explained 3.5–11.3% and 2.1–18.7% of the total genotypic variation, respectively. In contrast, the majority of QTLs detected were unique to only one environment, a finding which is consistent with QTL mapping studies of other forest trees, indicating a substantial contribution of environmental effects on the mapping progenies. Nonetheless, the two stable QTLs identified in this study could be important genomic regions to target for further research aimed at maximizing breeding efficiency and wood quality of C. japonica across wide environmental gradients.

PICEAdatabase: a web database for Picea omics and phenotypic information.

Picea belongs to the Pinaceae family and is a famous commercial tree species because of its straight trunk and excellent timber traits. Recently, omics have been widely used for fundamental and mechanism studies on Picea plants. To improve the accessibility to omics and phenotypic data and facilitate further studies, we compiled the sequences of 2 chloroplast genomes (Picea crassifolia and Picea asperata) and 32 complete omics data sets, including 20 transcriptomes, 4 proteomes, 2 degradomes and 6 microRNAs from P. crassifolia, P. asperata, Picea balfouriana and Picea abies tissues under different treatments, in PICEAdatabase. In addition, phenotypic data on plant growth and wood property traits were collected from two field trials of P. crassifolia. PICEAdatabase also includes useful analysis tools, such as BLAST, DESeq2 and JBrowse, to assist with analyses.

Genome-Wide Association Studies to Improve Wood Properties: Challenges and Prospects.

Wood formation is an excellent model system for quantitative trait analysis due to the strong associations between the transcriptional and metabolic traits that contribute to this complex process. Investigating the genetic architecture and regulatory mechanisms underlying wood formation will enhance our understanding of the quantitative genetics and genomics of complex phenotypic variation. Genome-wide association studies (GWASs) represent an ideal statistical strategy for dissecting the genetic basis of complex quantitative traits. However, elucidating the molecular mechanisms underlying many favorable loci that contribute to wood formation and optimizing GWAS design remain challenging in this omics era. In this review, we summarize the recent progress in GWAS-based functional genomics of wood property traits in major timber species such as Eucalyptus, Populus, and various coniferous species. We discuss several appropriate experimental designs for extensive GWAS in a given undomesticated tree population, such as omics-wide association studies and high-throughput phenotyping technologies. We also explain why more attention should be paid to rare allelic and major structural variation. Finally, we explore the potential use of GWAS for the molecular breeding of trees. Such studies will help provide an integrated understanding of complex quantitative traits and should enable the molecular design of new cultivars.

Genetic architecture underlying the lignin biosynthesis pathway involves noncoding RNAs and transcription factors for growth and wood properties in Populus.

SummaryLignin provides structural support in perennial woody plants and is a complex phenolic polymer derived from phenylpropanoid pathway. Lignin biosynthesis is regulated by coordinated networks involving transcription factors (TFs), microRNAs (miRNAs) and long noncoding RNAs (lncRNAs). However, the genetic networks underlying the lignin biosynthesis pathway for tree growth and wood properties remain unknown. Here, we used association genetics (additive, dominant and epistasis) and expression quantitative trait nucleotide (eQTN) mapping to decipher the genetic networks for tree growth and wood properties in 435 unrelated individuals of Populus tomentosa. We detected 124 significant associations (P ≤ 6.89E‐05) for 10 growth and wood property traits using 30 265 single nucleotide polymorphisms from 203 lignin biosynthetic genes, 81 TF genes, 36 miRNA genes and 71 lncRNA loci, implying their common roles in wood formation. Epistasis analysis uncovered 745 significant pairwise interactions, which helped to construct proposed genetic networks of lignin biosynthesis pathway and found that these regulators might affect phenotypes by linking two lignin biosynthetic genes. eQTNs were used to interpret how causal genes contributed to phenotypes. Lastly, we investigated the possible functions of the genes encoding 4‐coumarate: CoA ligase and cinnamate‐4‐hydroxylase in wood traits using epistasis, eQTN mapping and enzymatic activity assays. Our study provides new insights into the lignin biosynthesis pathway in poplar and enables the novel genetic factors as biomarkers for facilitating genetic improvement of trees.

The Interactions between the Long Non-coding RNA NERDL and Its Target Gene Affect Wood Formation in Populus tomentosa.

Long non-coding RNAs (lncRNAs) are important regulatory factors for plant growth and development, but little is known about the allelic interactions of lncRNAs with mRNA in perennial plants. Here, we analyzed the interaction of the NERD (Needed for RDR2-independent DNA methylation) Populus tomentosa gene PtoNERD with its putative regulator, the lncRNA NERDL (NERD-related lncRNA), which partially overlaps with the promoter region of this gene. Expression analysis in eight tissues showed a positive correlation between NERDL and PtoNERD (r = 0.62), suggesting that the interaction of NERDL with its putative target might be involved in wood formation. We conducted association mapping in a natural population of P. tomentosa (435 unrelated individuals) to evaluate genetic variation and the interaction of the lncRNA NERDL with PtoNERD. Using additive and dominant models, we identified 30 SNPs (P < 0.01) associated with five tree growth and wood property traits. Each SNP explained 3.90–8.57% of phenotypic variance, suggesting that NERDL and its putative target play a common role in wood formation. Epistasis analysis uncovered nine SNP-SNP association pairs between NERDL and PtoNERD, with an information gain of -7.55 to 2.16%, reflecting the strong interactions between NERDL and its putative target. This analysis provides a powerful method for deciphering the genetic interactions of lncRNAs with mRNA and dissecting the complex genetic network of quantitative traits in trees.

Identification and allelic dissection uncover roles of lncRNAs in secondary growth of Populus tomentosa.

Long non-coding RNAs (lncRNAs) function in various biological processes. However, their roles in secondary growth of plants remain poorly understood. Here, 15,691 lncRNAs were identified from vascular cambium, developing xylem, and mature xylem of Populus tomentosa with high and low biomass using RNA-seq, including 1,994 lncRNAs that were differentially expressed (DE) among the six libraries. 3,569 cis-regulated and 3,297 trans-regulated protein-coding genes were predicted as potential target genes (PTGs) of the DE lncRNAs to participate in biological regulation. Then, 476 and 28 lncRNAs were identified as putative targets and endogenous target mimics (eTMs) of Populus known microRNAs (miRNAs), respectively. Genome re-sequencing of 435 individuals from a natural population of P. tomentosa found 34,015 single nucleotide polymorphisms (SNPs) within 178 lncRNA loci and 522 PTGs. Single-SNP associations analysis detected 2,993 associations with 10 growth and wood-property traits under additive and dominance model. Epistasis analysis identified 17,656 epistatic SNP pairs, providing evidence for potential regulatory interactions between lncRNAs and their PTGs. Furthermore, a reconstructed epistatic network, representing interactions of 8 lncRNAs and 15 PTGs, might enrich regulation roles of genes in the phenylpropanoid pathway. These findings may enhance our understanding of non-coding genes in plants.

The interactive effect of root disease and climate on wood properties in halfsibling Douglas-fir families

Certain tree phenotypes enhance wood value properties and promote adaptation to climate extremes. However, the disciplines of wood science, physiology, and pathology rarely intersect to elucidate how survival, productivity, and wood property traits relate. Reaction to biotic and abiotic stress agents expressed in trees as disease resistance or tolerance were studied at the cell and tissue level. Five field-grown 21–22-year-old maternal halfsibling Douglas-fir Pseudotsuga menziesii var. glauca (Beissn.) Franco families were inoculated with Armillaria ostoyae (Romagn.) Herink to determine the interaction between disease, climate, and family on wood properties obtained from cores. The families originated from a lower elevation ecosystem and were categorized a priori as fungal disease resistant, tolerant or susceptible from a field study. Cell property changes suggestive of hydraulic adaptation to low precipitation were similar but lesser than the changes resulting from fungal root infection. The greatest increase in wood density occurred in disease resistant families after infection through a combination of thicker cell walls, smaller tracheid radial diameter, and reduced earlywood width. Microfibril angle and modulus of elasticity were affected differently with respect to changes in atmospheric relative humidity. Disease resistant compared to susceptible and tolerant families had inherent differences regardless of infection status. The sudden and localized changes in wood density associated with a disease resistance response could negatively impact on product quality, uniformity, and growth; on the other hand, denser wood and reduced growth associated with disease resistance may enhance drought survival. Understanding the interaction of these traits is important for adaptation to environmental stressors.