Japanese larch (Larix kaempferi) as a successful exotic species has become one of the most important economic and ecological conifers in China. In order to broaden the genetic resource of Larix kaempferi, an effort was made in 1996 to introduce 128 families from seven seed orchards in Japan, with which to establish two progeny trials in climatically different environments. The experiment was aimed to determine the strategy of early selection, particularly important for long-rotated Japanese larch, and the optimal breeding program for specific environments. Growth trajectories revealed different growth performances of stem height (HGT) and diameter at breast height (DBH) in two different environments, Hubei and Liaoning. In both sites, there were marked variabilities in HGT, DBH and volume (VOL) among families at each year. The trends of individual and family heritability and age-age correlations were found to follow a certain dynamic pattern. Based on these trends, the optimum selection age was determined at four years for HGT and five years for DBH in Hubei and Liaoning. Genetic gains for VOL were 34.4 and 6.04 % in Hubei and Liaoning respectively when selection ratio was 10 % at age 16. Type-B correlations were less than 0.67 and rank correlations of breeding value were less than 0.4 for HGT, DBH and VOL between the two sites, revealing that there exist pronounced family-by-site interactions for the growth traits of Larix kaempferi. Early selection for Larix kaempferi is an effective strategy to overcome its long rotation age. In early selection, dual growth trait selection is more effective than single one. Regionalization deployment should be considered in Larix. kaempferi breeding program based on different environmental factors.

Hybrids of larch (Larix kaempferi × Larix olgensis) are important afforestation species in northeastern China. They are routinely propagated via rooted stem cuttings. Despite the importance of rooting, little is known about the regulation of adventitious root development in larch hybrids. 454 GS FLX Titanium technology represents a new method for characterizing the transcriptomes of non-model species. This method can be used to identify differentially expressed genes, and then two-dimensional difference gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF/TOF MS) analyses can be used to analyze their corresponding proteins. In this study, we analyzed semi-lignified cuttings of two clones of L. kaempferi × L. olgensis with different rooting capacities to study the molecular basis of adventitious root development. We analyzed two clones; clone 25-5, with strong rooting capacity, and clone 23-12, with weak rooting capacity. We constructed four cDNA libraries from 25-5 and 23-12 at two development stages. Sequencing was conducted using the 454 pyrosequencing platform. A total of 957832 raw reads was produced; 95.07% were high-quality reads, and were assembled into 45137 contigs and 61647 singletons. The functions of the unigenes, as indicated by their Gene Ontology annotation, included diverse roles in the molecular functions, biological processes, and cellular component categories. We analyzed 75 protein spots (-fold change ≥2, P ≤0.05) by 2D-DIGE, and identified the differentially expressed proteins using MALDI-TOF/TOF MS. A joint analysis of transcriptome and proteome showed genes related to two pathways, polyamine synthesis and stress response, might play an important role on adventitious root development. These results provide fundamental and important information for research on the molecular mechanism of adventitious root development. We also demonstrated for the first time the combined use of two important technologies as a powerful approach to advance research on non-model, but otherwise important, larch species.

New simple sequence repeat (SSR) markers were developed in the Japanese larch (Larix kaempferi) using unigene sequences for further genetic diversity studies and the genetic improvement of breeding programs. One thousand two handred and thirty five (1235) primer pairs were tested and 165 successfully identified in L. kaempferi. Out of the amplified candidate markers, 145 (90.6%) exhibited polymorphism among 24 individuals of L. kaempferi, with the number of alleles per locus (N_a), observed heterozygosity (H_o), expected heterozygosity (H_e) and polymorphic information content (PIC) averaging at 4.510, 0.487, 0.518 and 0.459, respectively. Cross-species amplification of randomly selection of 30 genic-SSRs among the 145 polymorphic ones showed that 80.0% of the SSRs could be amplified in Larix olgensis, 86.7% could be amplified in Larix principi-rupprechtii and 83.0% could be amplified in Larix gmelinii. High rates of cross-species amplification were observed. The genic-SSRs developed herein would be a valuable resource for genetic analysis of Larix kaempferi and related species, and also have the potential to facilitate the genetic improvement and breeding of larch.

The increase in the atmospheric carbon dioxide concentration promotes its accumulation in trees by regulating the synthesis and transportation genes for endogenous hormones, such as IAA and GA, which are key factors in regulating various life activities, including growth rings. To explore the impact of changes in endogenous hormone levels such as IAA and GA on the growth of tree rings, and to provide a basis for improving the management of hybrid larch clonal forests, we investigated the effects of exogenous indole-3-acetic acid (IAA), gibberellic acid 3 (GA₃), and their combination on tree-ring growth in hybrid larch. IAA, GA₃, and a combination treatment were sprayed on the leaves of one clone of a hybrid larch seedling every three days. Small blocks were collected at the base stems for sequential anatomical observations. The phytohormone type, instead of the concentration, had a more significant effect on wood formation. Specifically, IAA treatment at 300 mg L⁻¹ significantly increased latewood (LW) layers until 90 days after treatment (DAT). The 500 mg L⁻¹ treatment significantly increased the wall radial thickness (WRT) of latewood (LW) cells. GA₃ treatment at 100 mg L⁻¹ significantly decreased the layers and width of total wood (TW), LW, and earlywood (EW). The 300 mg L⁻¹ treatment significantly increased the WRT of EW. The IAA 100 mg L⁻¹ + GA₃ 100 mg L⁻¹ combination treatment significantly increased the layers and width of TW and LW by inducing cambium activity and increasing the rate of wood cell development. The WRT and lumen radial diameter (LRD) of EW or LW in this treatment were similar to those observed with the corresponding single phytohormone treatment. These results indicate that combination treatment at 100 mg L⁻¹ + 100 mg L⁻¹ was a better way to promote tree-ring growth. Our study suggests that changes in phytohormone levels and ratios are important factors that affect tree-ring formation. Hormone levels and ratios should be regarded as important indicators to guide the improvement of management practices in hybrid larch clonal plantations.