Yoshitake Fujisawa

Transcriptome sequencing and profiling of expressed genes in cambial zone and differentiating xylem of Japanese cedar ( Cryptomeria japonica )

BackgroundForest trees have ecological and economic importance, and Japanese cedar has highly valued wood attributes. Thus, studies of molecular aspects of wood formation offer practical information that may be used for screening and forward genetics approaches to improving wood quality.ResultsAfter identifying expressed sequence tags in Japanese cedar tissue undergoing xylogenesis, we designed a custom cDNA microarray to compare expression of highly regulated genes throughout a growing season. This led to identification of candidate genes involved both in wood formation and later cessation of growth and dormancy. Based on homology to orthologous protein groups, the genes were assigned to functional classes. A high proportion of sequences fell into functional classes related to posttranscriptional modification and signal transduction, while transcription factors and genes involved in the metabolism of sugars, cell-wall synthesis and lignification, and cold hardiness were among other classes of genes identified as having a potential role in xylem formation and seasonal wood formation.ConclusionsWe obtained 55,051 unique sequences by next-generation sequencing of a cDNA library prepared from cambial meristem and derivative cells. Previous studies on conifers have identified unique sequences expressed in developing xylem, but this is the first comprehensive study utilizing a collection of expressed sequence tags for expression studies related to xylem formation in Japanese cedar, which belongs to a different lineage than the Pinaceae. Our characterization of these sequences should allow comparative studies of genome evolution and functional genetics of wood species.

Soft X-ray observation of water distribution in the stem of Cryptomeria japonica D. Don I: General description of water distribution

Water distribution in green stems ofCryptomeria japonica D. Don was observed by soft X-ray photography. In the sapwood, much water was present and evenly distributed. In the intermediate wood (the white zone), little water was present. The intermediate wood appeared in all cross sections of the stem and separated the heartwood from the sapwood in the intertracheid water connection. Maldistribution of water was generally observed in the heartwood, and three types of water presence were distinguishable: a “wet area” with accumulated water, a “dry area” with little water, and a “moderate moisture area” with intermediate accumulation. The distribution pattern and amount of water in the heartwood varied dramatically among and even within trees. Separation of the heartwood from the sapwood in the intertracheid water connection suggested that the presence of water in the heartwood was caused by rewetting of the tracheid lamina that occurred after heartwood formation. The maldistribution of water in the heartwood suggested that a difference in the process of rewetting causes both uneven distribution and the various types of water presence.

Effects of Clonal Selection by Microfibril Angle on the Genetic Improvement of Stiffness in Cryptomeria japonica D. Don

Summary The microfibril angle (MFA) of latewood tracheids and its radial variation at breast height in Cryptomeria japonica D. Don (sugi) were investigated with twelve clones collected from three sites in the Kyushu region, Japan. Large variations both between clones and between sites were observed. The MFAs were well correlated to the stiffness of the logs collected from the sample trees. A simulation of clonal selection according to the ranking of the clones in MFA demonstrated that the log stiffness of the selected population was much improved even when the selection relied on MFAs in the second ring from the pith. The improvement in log stiffness by MFA selection was not different from the selection by log stiffness itself. The results indicate that early selection by MFA is very effective in improving log stiffness in this species.

Soft X-ray observation of water distribution in the stem of Cryptomeria japonica D. Don II: Types found in wet-area distribution patterns in transverse sections of the stem

Using soft X-ray photography, two-dimensional observations were made of the water distribution within the green stem ofCryptomeria japonica D. Don. Variations in the distribution pattern of the “wet area” in the horizontal plane of the lower stem were described. The distribution patterns showed extreme variation among individual trees, with broad wet-area distribution types appearing, namely, the regularly distributed wet area and the irregularly distributed wet area. We defined five basic types of wet-area distribution patterns on the basis of their regularity or irregularity. It was concluded that the between-tree variation in the wet-area distribution causes the between-tree variation in the mean moisture content of the heartwood. The distribution patterns of the wet area were similar for individual trees within each plus-tree clone.

Changes in water distribution in heartwood along stem axes in Cryptomeria japonica

Abstract Three-dimensional distribution of water in the heartwood of Cryptomeria japonica D. Don was observed by soft X-ray photography. The within-tree variation in the distribution of “wet areas” (water-accumulated areas in heartwood) was enormous, as was the variation among trees. Although we found no universal pattern of changes in wet area distributions along the stem axes of all trees, similarities among individual trees within each cultivar and clone were observed. The difference between the two kinds of wetwood in C. japonica – genetically defined wetwood and secondarily induced wetwood – is discussed, as is the use of soft X-ray photography in Japanese tree breeding programs.

Microfibril angle and density of hinoki (Chamaecyparis obtusa) trees in 15 half-sib families in a progeny test stand in Kyushu, Japan

It was previously believed in Japan that the wood qualities of hinoki (Chamaecyparis obtusa) were superior to sugi (Cryptomeria japonica). However, few studies of wood properties such as MFA (microfibril angle of S2 layer in secondary wall of tracheid) have been completed for hinoki. Some reports have found that hinoki plus tree families have similar mechanical properties to sugi. Here we report the characteristics of MFA and density of hinoki half-sib families in a progeny test stand. There were significant differences in MFA and density between families. The wood properties of two families, Nakatsu 3 and Kanzaki 5, are stable in radial pattern and suitable for structural use. Early selection of hinoki families by MFA and density may be difficult. Effects of MFA and density on Ed (dynamic modulus of elasticity) of logs differed between families. The effects of growth rate on MFA and density differed between families and also between juvenile and mature wood. The faster growth rate in Nakatsu 3 appeared to improve wood properties and increase Ed of logs, although in many other families, faster growth rate had negative effects on desirable wood properties for structural use.

Termite resistance and color of heartwood of hinoki (Chamaecyparis obtusa) trees in 5 half-sib families in a progeny test stand in Kyushu, Japan

Hinoki (Chamaecyparis obtusa) wood has been used as a structural material in Japan because of its superior mechanical properties, its excellent durability and the beautiful color of its heartwood. Variations of termite resistance and compositions of extractives among hinoki trees have been reported. However, genetic variation of termite resistance and the effect of heartwood color on termite resistance remain unknown. In this study, we report the characteristics of termite resistance and color indexes (L*, a* and b*) of heartwood of hinoki half-sib families in a progeny test stand. The survival days of termites and the mass loss of samples of hinoki heartwood differed significantly among hinoki families. Families with red-color heartwood had larger termite resistance than families with yellow-color heartwood. The termite resistances of individual samples from two families with yellow-color heartwood were as small as that of Pinus densiflora. Larger a* and smaller b* induced larger termite resistance of heartwood. The effect of DBH (diameter at breast height) of hinoki trees on termite resistance of hinoki families was small.

Species characteristics and intraspecific variation in growth and photosynthesis of Cryptomeria japonica under elevated O 3 and CO 2

In order to predict the effects of future atmospheric conditions on forest productivity, it is necessary to clarify the physiological responses of major forest tree species to high concentrations of ozone (O3) and carbon dioxide (CO2). Furthermore, intraspecific variation of these responses should also be examined in order to predict productivity gains through tree improvements in the future. We investigated intraspecific variation in growth and photosynthesis of Cryptomeria japonica D. Don, a major silviculture species in Japan, in response to elevated concentrations of O3 (eO3) and CO2 (eCO2), separately and in combination. Cuttings of C. japonica were grown and exposed to two levels of O3 (ambient and twice-ambient levels) in combination with two levels of CO2 (ambient and 550 µmol mol-1 in the daytime) for two growing seasons in a free-air CO2 enrichment experiment. There was no obvious negative effect of eO3 on growth or photosynthetic traits of the C. japonica clones, but a positive effect was observed for annual height increments in the first growing season. Dry mass production and the photosynthetic rate increased under eCO2 conditions, while the maximum carboxylation rate decreased. Significant interaction effects of eO3 and eCO2 on growth and photosynthetic traits were not observed. Clonal effects on growth and photosynthetic traits were significant, but the interactions between clones and O3 and/or CO2 treatments were not. Spearmans rank correlation coefficients between growth traits under ambient conditions and for each treatment were significantly positive, implying that clonal ranking in growth abilities might not be affected by either eO3 or eCO2. The knowledge obtained from this study will be helpful for species selection in afforestation programs, to continue and to improve current programs involving this species, and to accurately predict the CO2 fixation capacity of Japanese forests.

Erratum to: Transcriptome sequencing and profiling of expressed genes in cambial zone and differentiating xylem of Japanese cedar (Cryptomeria japonica)[BMC Genomics, 15, (2014) (219)], DOI: 10.1186/1471-2164-15-219

Background: Forest trees have ecological and economic importance, and Japanese cedar has highly valued wood attributes. Thus, studies of molecular aspects of wood formation offer practical information that may be used for screening and forward genetics approaches to improving wood quality. Results: After identifying expressed sequence tags in Japanese cedar tissue undergoing xylogenesis, we designed a custom cDNA microarray to compare expression of highly regulated genes throughout a growing season. This led to identification of candidate genes involved both in wood formation and later cessation of growth and dormancy. Based on homology to orthologous protein groups, the genes were assigned to functional classes. A high proportion of sequences fell into functional classes related to posttranscriptional modification and signal transduction, while transcription factors and genes involved in the metabolism of sugars, cell-wall synthesis and lignification, and cold hardiness were among other classes of genes identified as having a potential role in xylem formation and seasonal wood formation. Conclusions: We obtained 55,051 unique sequences by next-generation sequencing of a cDNA library prepared from cambial meristem and derivative cells. Previous studies on conifers have identified unique sequences expressed in developing xylem, but this is the first comprehensive study utilizing a collection of expressed sequence tags for expression studies related to xylem formation in Japanese cedar, which belongs to a different lineage than the Pinaceae. Our characterization of these sequences should allow comparative studies of genome evolution and functional genetics of wood species.