From Genome Sequencing to CRISPR-Based Genome Editing for Climate-Resilient Forest Trees

Due to the economic and ecological importance of forest trees, modern breeding and genetic manipulation of forest trees have become increasingly prevalent. The CRISPR-based technology provides a versatile, powerful, and widely accepted tool for analyzing gene function and precise genetic modification in virtually any species but remains largely unexplored in forest species. Rapidly accumulating genetic and genomic resources for forest trees enabled the identification of numerous genes and biological processes that are associated with important traits such as wood quality, drought, or pest resistance, facilitating the selection of suitable gene editing targets. Here, we introduce and discuss the latest progress, opportunities, and challenges of genome sequencing and editing for improving forest sustainability.

Differences in wood properties among Eucalyptus grandis and Eucalyptus grandis x Eucalyptus urophylla with different degrees of ploidy

We compared the anatomy, density, chemical contents, and bioenergy values of Eucalyptus grandis and hybrids of Eucalyptus grandis x Eucalyptus urophylla wood originating from diploids, triploids and tetraploids. We hypothesize that Eucalyptus grandis and hybrids of Eucalyptus grandis x Eucalyptus urophylla with different degrees of ploidy have variations as a result of different sets of chromosomes producing different phenotypic expressions and chemical constituents, such as variation in cell size and frequency, which would directly influence wood quality. Twenty-year-old trees were cut, eight for each ploidy: diploids and tetraploids are E. grandis; triploids are E. grandis x E. urophylla. We use standardized techniques. Our hypothesis was confirmed. Triploid and tetraploid trees presented wider trunks, taller trees with longer stems and wider crowns compared to diploid trees. Wood density showed significant radial variation only in diploids, while triploid and tetraploid trees were more homogeneous. In polyploid trees, the anatomical features did not clearly present a radial pattern. Triploid and tetraploid trees presented higher density wood than diploid trees. The chemical constituents varied from pith to bark in the three ploidies, but no differences between ploidies were found. For energy generation purposes, diploid and triploid trees are more desirable than tetraploid trees.

A Talk between Flavonoids and Hormones to Reorient the Growth of Gymnosperms

Plants reorient the growth of affected organs in response to the loss of gravity vector. In trees, this phenomenon has received special attention due to its importance for the forestry industry of conifer species. Sustainable management is a key factor in improving wood quality. It is of paramount importance to understand the molecular and genetic mechanisms underlying wood formation, together with the hormonal and environmental factors that affect wood formation and quality. Hormones are related to the modulation of vertical growth rectification. Many studies have resulted in a model that proposes differential growth in the stem due to unequal auxin and jasmonate allocation. Furthermore, many studies have suggested that in auxin distribution, flavonoids act as molecular controllers. It is well known that flavonoids affect auxin flux, and this is a new area of study to understand the intracellular concentrations and how these compounds can control the gravitropic response. In this review, we focused on different molecular aspects related to the hormonal role in flavonoid homeostasis and what has been done in conifer trees to identify molecular players that could take part during the gravitropic response and reduce low-quality wood formation.

Timber Properties and Cellulose Crystallites Size in Pine Wood Cut in Different Sawing Patterns after Pretreatment with CH3COOH and H2O2 and Densification

One process to improve wood quality is densification or wood surface compression. Our study analyzed the changes in some basic properties of pine wood, including its anatomical structure, density, modulus of elasticity (MOE), and dimensions of cellulose crystallites, after densification following soaking pretreatment in CH3COOH and H2O2 at a concentration of 20%. Samples were sawn in radial and tangential directions for analysis of the wood. The results showed a change in the shape of tracheid cells from hexagonal to oval, as well as damage to the ray cell constituents on the tangential surface. The thickness decrease of the samples was in accordance with the target, which meant that spring-back was short. In general, the tangential boards had a higher density than the radial boards, with a lower MOE and crystallite dimensions. Our findings showed that the densified tangential board was stronger than the radial board.

Seed Germination of Enterolobium contortisiliquum (Vell.) Morong. (Fabaceae) under Different Temperatures and Drying Method

Enterolobium contortisiliquum (Vell.) Morong a tree commonly known as anglerfish, monkey ear and timbaúva. Due to its wood quality, this tree species is widely used in naval and civil constructions, toys and furniture frames, and can also be used for urban afforestation and restoration of degraded areas. The objective of this work was to standardize the best temperature conditions for conducting the germination tests of E. contortisiliquum seeds under laboratory conditions, considering the fact that the seeds come from coastal tropical Brazil, and to evaluate the desiccation tolerance. The experiments were carried out at the Plant Propagation Laboratory of the Agricultural Engineering and Sciences Campus (CECA) of the Federal University of Alagoas (UFAL), located in the municipality of Rio Largo, Alagoas, Brazil. The design of the experiment was fully randomized, with four repetitions of 25 seeds per treatment. In test I: The newly harvested seeds were subjected to constant temperatures of 5, 10, 15, 20, 25, 30, 35 and 40ºC and alternating temperatures of 20-30ºC, with an eight-hour photoperiod, simulated by four fluorescent lamps 20W daylight type. In trial II: The seeds were divided into two lots. The first was the control (initial moisture content). The second was subjected to slow drying on silica gel. A constant temperature of 30ºC is recommended for the germination and vigor test of E. contortisiliquum seeds. The critical means water content for seeds of this species is below 5%.

Transcriptome and microRNA Sequencing Identified miRNAs and Target Genes in Different Developmental Stages of the Vascular Cambium in Cryptomeria fortunei Hooibrenk

Cryptomeria fortunei Hooibrenk is an important fast-growing coniferous timber species that is widely used in landscaping. Recently, research on timber quality has gained substantial attention in the field of tree breeding. Wood is the secondary xylem formed by the continuous inward division and differentiation of the vascular cambium; therefore, the development of the vascular cambium is particularly important for wood quality. In this study, we analyzed the transcriptomes of the cambial zone in C. fortunei during different developmental stages using Illumina HiSeq sequencing, focusing on general transcriptome and microRNA (miRNA) data. We performed functional annotation of the differentially expressed genes (DEGs) in the different stages identified by transcriptome sequencing and generated 15 miRNA libraries yielding 4.73 Gb of clean reads. The most common length of the filtered miRNAs was 21nt, accounting for 33.1% of the total filtered reads. We annotated a total of 32 known miRNA families. Some miRNAs played roles in hormone signal transduction (miR159, miR160, and miR166), growth and development (miR166 and miR396), and the coercion response (miR394 and miR395), and degradome sequencing showed potential cleavage sites between miRNAs and target genes. Differential expression of miRNAs and target genes and functional validation of the obtained transcriptome and miRNA data provide a theoretical basis for further elucidating the molecular mechanisms of cellular growth and differentiation, as well as wood formation in the vascular cambium, which will help improve the wood quality of C. fortunei.

Dimensional Stability of Treated Sengon Wood by Nano-Silica of Betung Bamboo Leaves

Sengon (Falcataria moluccana Miq.) is one of the fastest growing wood that is broadly planted in Indonesia. Sengon wood has inferior wood properties, such as a low density and dimensional stability. Therefore, sengon wood requires a method to improve its wood quality through wood modification. One type of wood modification is wood impregnation. On the other hand, Betung Bamboo leaves are considered as waste. Betung Bamboo leaves contain silica. Based on several researches, nano-SiO2 could improve fast-growing wood qualities. According to its perfect solubility in water, monoethylene glycol (MEG) is used in the study. The objectives are to evaluate the impregnation treatment (MEG and nano-silica originated from betung bamboo leaves) in regard to the dimensional stability and density of 5-year-old sengon wood and to characterize the treated sengon wood. MEG, MNano-Silica 0.5%, MNano-Silica 0.75%, and MNano-Silica 1% were used as impregnation solutions. The impregnation method was started with 0.5 bar of vacuum for 60 min, followed by 2.5 bar of pressure for 120 min. The dimensional stability, density, and characterization of the samples were studied through the use of scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy, X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). The results show that the treatment had a significant effect on the dimensional stability and density of sengon wood. Alterations in the morphology of treated sengon wood were observed through the full coverage of the pits on the vessel walls (SEM analysis results) and the detection of ethylene (FTIR analysis results) and silica (XRD and FTIR analysis results). Overall, the 0.75% MNano-Silica treatment was the most optimal treatment for increasing the dimensional stability and density of 5-year-old sengon wood.

Genome-Wide Identification of Long Non-Coding RNAs and Their Potential Functions in Poplar Growth and Phenylalanine Biosynthesis

Poplar is an important bioenergy tree species. lncRNAs play important roles in various biological regulatory processes, and their expression pattern is more tissue-specific than mRNAs. In this study, P. deltoides “Danhong” (Pd) and P. simonii “Tongliao1” (Ps) with different growth rates and wood quality were used as experimental materials, and the transcriptomes of their shoot apical meristem, xylem, and phloem were sequenced. Furthermore, high-throughput RNA sequencing analysis revealed that the expression patterns of genes and lncRNAs are different between the two genotypes. 6,355 lncRNAs were identified. Based on target prediction, lncRNAs and target genes were involved in ADP binding, oxidoreductase activity, phenylpropanoid biosynthesis, and cyanoamino acid metabolism. The DElncRNAs in two poplars were co-expressed with transcription factors and structural genes of lignin and flavonoid pathways. In addition, we found the potential target lncRNAs of miRNA. This result provides basic evidence for a better understanding of the regulatory role of lncRNAs in regulating phenylalanine molecular pathways and wood formation.

Mahogany (Swietenia macrophylla King.) as urban tree: tree growth and wood quality variation in a progeny test

Mahogany (Swietenia macrophylla) is a tree species widely planted not only for timber production but also as a shade tree in urban areas both on the side of the highway (green belt) and other green open areas. The objective of this study was to analyze the genetic variation of growth and wood quality of a mahogany progeny test at the Parung Panjang, Bogor, which will be converted into a seedling seed orchard for high quality seed production. The progeny test was established using a randomized complete block design involving 96 families originating from seven populations (land races) in Java Island, i.e., Cianjur, Sumedang, Tasikmalaya, Magelang, Gundih, Madiun, and Jember. The results showed that the family had a significant effect on all the growth and wood quality parameters. Individual and family heritability for all growth and wood quality traits were categorized into the high, i.e., 0.33 - 0.96 and 0.35 - 0.56, respectively. The selection simulation showed that the selection percentage of 44.68% (45 best families) gave the optimum genetic gain based on growth and wood quality traits.