Yuya Takashima

The breeding of a fast-growing tree species, Acacia mangium Willd. has entered the third-generation cycle in Indonesia. To fulfill timber industry needs, wood traits, as well as growth traits, should be improved by conducting appropriate tree breeding programs for this species. In the present study, growth and wood traits were measured for the 10-year-old third-generation A. mangium from 40 half-sib families originating from four sub-lines from two provenances. The mean values of ten families in each sub-line ranged from 17.7 to 21.8 cm for stem diameter, 14.3 to 16.2 m for tree height, 3.60 to 3.77 km/s for stress-wave velocity, and 19.9 to 20.9 mm for Pilodyn penetration, respectively. The broad-sense heritability values in each sub-line ranged from 0.014 to 0.444 for stem diameter, 0.000 to 0.208 for tree height, 0.261 to 0.715 for stress-wave velocity, and 0.171 to 0.484 for Pilodyn penetration, respectively. These results suggested that families with higher density and Young’s modulus of wood, suitable for solid wood production, can be selected from the third-generation A. mangium. Stem diameter significantly correlated with the other three measured traits, while phenotypic correlation coefficients between stem diameter and stress-wave velocity or Pilodyn penetration were very low. In addition, no significant phenotypic correlations were found between other measured traits. Furthermore, positive- or non-significant genetic correlations were found between growth and wood traits in all sub-lines. Thus, it is concluded that the selection of families with good growth characteristics will not always reduce the wood properties of this species.

As climate change progresses, the breeding of drought-tolerant forest trees is necessary. Breeding drought-tolerant trees requires screening for drought stress using a large number of individuals and a high-throughput phenotyping method. The aim of this study was therefore to establish high-throughput methods for evaluating the clonal stress responses to drought stress using infrared thermography and chlorophyll fluorescence methods in Cryptomeria japonica. The stomatal conductance index (Ig), maximum photochemical quantum yield of photosystem II (Fv/Fm), and axial growth of four plus-tree clones of C. japonica planted in pots were measured weekly for 85 days after irrigation was stopped. The phenotypic trait responsivity to drought stress was estimated by a nonlinear mixed model and by introducing the cumulative water index, which considers the past history of the soil water environment. These methods and procedures enabled us to evaluate the clonal stress responses in C. japonica and could be applied to large-scale clone materials to promote the breeding program for drought tolerance.

Along with progress in sequencing technology and accumulating knowledge of genome and gene sequences, molecular breeding techniques have been developed for predicting the genetic potential of individual genotypes and for selecting superior individuals. For Japanese cedar (Cryptomeria japonica (L.f.) D.Don), which is the most common coniferous species in Japanese forestry, we constructed a custom primer panel for target amplicon sequencing in order to simultaneously determine 3034 informative single nucleotide polymorphisms (SNPs). We performed primary evaluation of the custom primer panel with actual sequencing and in silico PCR. Genotyped SNPs had a distribution over almost the entire region of the C. japonica linkage map and verified the high reproducibility of genotype calls compared to SNPs obtained by genotyping arrays. Genotyping was performed for 576 individuals of the F₁ population, and genomic prediction models were constructed for growth and wood property-related traits using the genotypes. Amplicon sequencing with the custom primer panel enables efficient obtaining genotype data in order to perform genomic prediction, manage clones, and advance forest tree breeding.

To clarify inheritance of static bending properties in relation to elastic and plastic regions, air-dry density, microfibril angle of the S₂ layer in latewood tracheid (MFA), and static bending properties (modulus of elasticity [MOE], modulus of rupture [MOR] and bending work) were examined for juvenile wood of 18 full-sib families in 20-year-old Cryptomeria japonica. Heritability of all traits ranged from 0.12 (bending work) to 0.51 (air-dry density). Based on the results from principal component analysis (PCA) and cluster analysis, the families were classified into four groups with different types of load-deflection curves, suggesting that both elastic properties and deflection in plastic region differed among families. Furthermore, families included in a group were produced from specific parents, suggesting that deflection in plastic region as well as elastic property is inheritable. It can be concluded that mating parents may affect elasticity and plasticity of offspring.

The plant hormone gibberellin (GA) is known to regulate elongating growth, seed germination, and the initiation of flower bud formation, and it has been postulated that GAs originally had functions in reproductive processes. Studies on the mechanism of induction of flowering by GA have been performed in Arabidopsis and other model plants. In coniferous trees, reproductive organ induction by GAs is known to occur, but there are few reports on the molecular mechanism in this system. To clarify the gene expression dynamics of the GA induction of the male strobilus in Cryptomeria japonica, we performed comprehensive gene expression analysis using a microarray. A GA-treated group and a nontreated group were allowed to set, and individual trees were sampled over a 6-week time course. A total of 881 genes exhibiting changed expression was identified. In the GA-treated group, genes related to ‘stress response’ and to ‘cell wall’ were initially enriched, and genes related to ‘transcription’ and ‘transcription factor activity’ were enriched at later stages. This analysis also clarified the dynamics of the expression of genes related to GA signaling transduction following GA treatment, permitting us to compare and contrast with the expression dynamics of genes implicated in signal transduction responses to other plant hormones. These results suggested that various plant hormones have complex influences on the male strobilus induction. Additionally, principal component analysis (PCA) using expression patterns of the genes that exhibited sequence similarity with flower bud or floral organ formation-related genes of Arabidopsis was performed. PCA suggested that gene expression leading to male strobilus formation in C. japonica became conspicuous within one week of GA treatment. Together, these findings help to clarify the evolution of the mechanism of induction of reproductive organs by GA.

The present study was performed to unravel the mechanisms of systemic acquired resistance (SAR) establishment and resistance signaling pathways against the canker-rot fungus (Inonotus obliquus strain IO-U1) infection in Japanese birch plantlet No.8. Modulation of protein-profile induced by salicylic acid (SA)-administration was analyzed, and SA-responsive proteins were identified. In total, 5 specifically expressed, 3 significantly increased, and 3 significantly decreased protein spots were identified using liquid chromatography/tandem mass spectrometry (LC/MS/MS) and the sequence tag method. These proteins were malate dehydrogenase, succinate dehydrogenase, phosphoglycerate kinase, diaminopimalate decarboxylase, arginase, chorismate mutase, cyclophilin, aminopeptidase, and unknown function proteins. These proteins are considered to be involved in SAR-establishment mechanisms in the Japanese birch plantlet No 8.