Large ex situ germplasm collections of plants generally contain significant diversity. A set of 700 well-conserved Chinese fir (Cunninghamia lanceolata (Lamb.) Hook) clones from six provinces in southern China in the ex situ gene bank of Longshan State Forest, was analyzed using 21 simple sequence repeat markers, with the aim of assessing the genetic diversity of these germplasm resources. Genetic analysis revealed extensive genetic variation among the accessions, with an average of 8.31 alleles per locus and a mean Shannon index of 1.331. Excluding loci with null alleles, we obtained a low level of genetic differentiation among provinces, consistent with the interpopulation genetic variation (1%). Three clusters were identified by STRUCTURE, which did not match the individuals’ geographical provenances. Ten traits related to growth and wood properties were quantified in these individuals, and there was substantial variation in all traits across individuals, these provide a potential source of variation for genetic improvement of the Chinese fir. Screening large collections for multiple-trait selective breeding programs is laborious and expensive; a core collection of 300 accessions, representative of the germplasm, was established, based on genotypic and phenotypic data. The identified small, but diverse, collections will be useful for further genome-wide association studies.

To determine the phenotypic variation in 700 ten-year grafted Chinese fir collected from six provinces in southern China, 10 phenotypic traits were investigated: tree height, diameter at breast height, bark thickness, volume of timber, heartwood ratio, density of wood, hygroscopicity, tracheid length, tracheid diameter, and ratio of tracheid length to tracheid diameter. Abundant phenotypic variation was found among the six populations; the phenotypic variation coefficients all exceeded 10%, and the largest was for volume of timber. Significant variation (p < 0.01 or 0.05) in traits was found among the populations, except for diameter at breast height, heartwood ratio, and tracheid diameter, while all traits differed significantly (p < 0.01) within populations. The high value of repeatability (broad-sense heritability) suggested moderate genetic control of the traits. The 10 traits were strongly correlated within the entire population; strong positive correlations (p < 0.01) were observed between growth traits, and significant negative correlations (p < 0.01 or 0.05) were found between the density of wood and most other characteristics, except for heartwood ratio and ratio of tracheid length to tracheid diameter. Using diameter at breast height and density of wood as criteria, 98 relatively fast-growing genotypes with relatively high wood basic density were identified.

The tree species Pinus tabulaeformis Carr. (P. tabulaeformis) is commonly planted in China due to its economic and ecological value. In order to identify one or more ectomycorrhizal (ECM) fungal species for future P. tabulaeformis afforestation, we investigated the effects of five ECM fungal species: Laccaria laccata, Boletus edulis, Gomphidius viscidus, Suillus grevillei, and Suillus luteus on the growth of P. tabulaeformis seedlings under greenhouse conditions. The growth parameters of P. tabulaeformis seedlings were evaluated 90 days following fungal colonisation. The majority of seedlings were significantly affected by ECM inoculation. Mycorrhizal inoculated seedlings were taller, had more lateral roots, and a greater biomass compared with the non-mycorrhizal (CK) seedlings. With the exception of G. viscidus, inoculated seedlings exhibited higher phosphorus, potassium, and nitrogen content compared with the CK seedlings. In addition, ECM colonisation increased the enzymatic activity of catalase, acidic phosphatase, protease, and the urease content in the rhizosphere soil. Our study showed that Laccaria laccata, Suillus grevillei, and Suillus luteus may be useful for improving the growth and cultivation of P. tabulaeformis seedlings. Furthermore, we observed that S. luteus inoculation increased the gas exchange parameters of P. tabulaeformis seedlings under field conditions.

The chemical composition of secondary metabolites is important for the quality control of wood products. In this study, the widely targeted metabolomics approach was used to analyze the metabolic profiles of heartwood and sapwood in the red-heart Chinese fir (Cunninghamia lanceolata), with an ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometry system. A total of 224 secondary metabolites were detected in the heartwood and sapwood, and of these, flavonoids and phenolic acids accounted for 36% and 26% of the components, respectively. The main pathways appeared to be differentially activated, including those for the biosynthesis of phenylpropanoids and flavonoids. Moreover, we observed highly significant accumulation of naringenin chalcone, dihydrokaempferol, pinocembrin, hesperetin, and other important secondary metabolites in the flavonoid biosynthesis pathway. Our results provide insight into the flavonoid pathway associated with wood color formation in Chinese fir that will be useful for further breeding programs.

Robinia pseudoacaciais an important afforestation tree introduced to China in 1878. In the present study, we examined the genetic diversity among 687 strains representing four improved varieties and two secondary provenances, comprising 641 clones and 46 seedlings. Ninety-one simple sequence repeats (SSRs) were selected through segregation analysis and polymorphism characterization, and all sampled individuals were genotyped using well-characterized SSR markers. After excluding loci with non-neutral equilibrium, missing locus data and null alleles, we used 36 primer pairs to assess the genetic diversity of these germplasm resources, revealing vast genetic differentiation among the samples, with an average of 8.352 alleles per locus and a mean Shannon′s index of 1.302. At the population level, the partitioning of variability was assessed using analysis of molecular variance, which revealed 93% and 7% variation within and among collection sites, respectively. Four clusters were detected usingstructureanalysis, indicating a degree of genetic differentiation among the six populations. Insights into the genetic diversity and structure ofR. pseudoacaciaprovide a theoretical basis for the conservation, breeding and sustainable development in China.

Red-heart Chinese fir (Cunninghamia lanceolata) has the advantages of high density and attractive color, making it popular in the market. To date, most studies about stems of woody plants have only been reported at the cytological level because of few living cells. In this study, the xylem was successfully partitioned into three effective sampling areas: sapwood, transition zone, and heartwood. Secondary metabolites, cell survival, and differentially expressed genes in the three sampling areas were, respectively, investigated. First, we identified the phenylpropanoid and flavonoid pathways closely related to color. Based on the chemical structure of secondary metabolites in pathways, two notable directions had been found. Luteolin’s glycosylation products might be the key substances that regulated the color of heartwood in red-heart Chinese fir because of the 1,000-fold difference between red-heart and white-heart. We also found pinocembrin and pinobanksin in Chinese fir, which were rarely reported before. At the cytological level, we believed that the transition zone of red-heart Chinese fir was a critical region for color production because of the fewer living ray parenchyma cells. In addition, transcriptome and quantitative reverse transcription PCR (qRT-PCR) proved that genes regulating the entire phenylpropanoid pathway, upstream of the flavonoid pathway, and some glycosyltransferases were significantly upregulated in the transition zone of red-heart and then colored the heartwood by increasing metabolites. This is the first report on the color-related secondary metabolites regulated by differential genes in red-heart Chinese fir. This study will broaden our knowledge on the effects of metabolites on coloring woody plant xylems.