Light Competition Contributes to the Death of Masson Pines of Coniferous-Broadleaf Mixed Forests in Subtropical China

In the process of subtropical forest succession, it has long been recognized that population decline of Masson pines in coniferous-broadleaf mixed forest is caused by shading from broadleaf trees. However, little is known about the mechanism underlying the interaction between them. Here, we first chose two sets of Masson pine plots approximately aged 60 years in subtropical mountainous areas in eastern China (i.e., pure coniferous forest vs. coniferous-broadleaf mixed forest). Then, we measured and compared tree height, diameter at breast height, first branch height (FBH), live crown ratio (LCR) of Masson pines between the two sets of plots, and also determined the difference in growth performance of Masson pines relative to their neighboring broadleaf trees in the mixed forest stand. Compared with plots in pine forests, Masson pines in mixed plots had lower tree height and crown breadth, higher FBH, lower LCR, and leaf area. Furthermore, the difference of mean FBH between reference trees (Masson pines) and their neighboring trees (i.e., broadleaf trees) in mixed forest plots was greater than that in pine forest plots, and the ratio of LCR between Masson pines and their neighbors (0.46) in mixed forest was significantly smaller than in pine forest (1.05), indicating that those broadleaf trees around Masson pines probably affected their growth. The mean distance between Masson pines and neighboring trees (1.59 m) in mixed forest plots was significantly shorter than in pine forest plots (2.77 m) (p < 0.01), suggesting that strong competition may occur between reference trees and their neighbors. There was a significant difference in the ratio of crown volume between reference tree Masson pine and its neighboring trees in mixed forests (p < 0.01), indicating that the ratio of biomass synthesis to consumption of pines was much lower than their nearby broadleaf trees in mixed forest. Our results have demonstrated for the first time that Masson pines’ population decline is affected by shade-tolerant broadleaf late-successional species, which can be primarily attributed to the distinctive light transmittance of dominant species nearby (pure pine vs. mixed forest). This study provides a new perspective for future studies on the mechanism of forest succession.

Multi-Omics of Pine Wood Nematode Pathogenicity Associated With Culturable Associated Microbiota Through an Artificial Assembly Approach

Pinewood nematode (PWN), the causal agent of pine wilt disease (PWD), causes massive global losses of Pinus species each year. Bacteria and fungi existing in symbiosis with PWN are closely linked with the pathogenesis of PWD, but the relationship between PWN pathogenicity and the associated microbiota is still ambiguous. This study explored the relationship between microbes and the pathogenicity of PWN by establishing a PWN-associated microbe library, and used this library to generate five artificial PWN–microbe symbiont (APMS) assemblies with gnotobiotic PWNs. The fungal and bacterial communities of different APMSs (the microbiome) were explored by next-generation sequencing. Furthermore, different APMSs were used to inoculate the same Masson pine (Pinus massoniana) cultivar, and multi-omics (metabolome, phenomics, and transcriptome) data were obtained to represent the pathogenicity of different APMSs at 14 days post-inoculation (dpi). Significant positive correlations were observed between microbiome and transcriptome or metabolome data, but microbiome data were negatively correlated with the reactive oxygen species (ROS) level in the host. Five response genes, four fungal genera, four bacterial genera, and nineteen induced metabolites were positively correlated with the ROS level, while seven induced metabolites were negatively correlated. To further explore the function of PWN-associated microbes, single genera of functional microbes (Mb1–Mb8) were reloaded onto gnotobiotic PWNs and used to inoculate pine tree seedlings. Three of the genera (Cladophialophora, Ochroconis, and Flavobacterium) decreased the ROS level of the host pine trees, while only one genus (Penicillium) significantly increased the ROS level of the host pine tree seedlings. These results demonstrate a clear relationship between associated microbes and the pathogenicity of PWN, and expand the knowledge on the interaction between PWD-induced forest decline and the PWN-associated microbiome.

Variability in Soil Macronutrient Stocks across a Chronosequence of Masson Pine Plantations

Plantations play a vital role in the global nutrient cycle because they have large stocks of soil macronutrients. However, the impacts of plantations on soil macronutrient stocks combined with stand age and soil physicochemical properties have not been well quantified. We compared soil macronutrient stocks at soil depths of 0−20 and 20−40 cm across a 7-, 14-, 25-, and 30-year chronosequence of Masson pine (Pinus massoniana Lamb.) plantations. The results showed that the nitrogen (N), phosphorus (P), and potassium (K) stocks first increased and then decreased with stand age. The highest N and P stocks were observed in the 14-year-old plantation, and the 25-year-old plantation displayed the highest K stock. The C, N, and P stocks declined with increasing soil depth across all sites, whereas the reverse trend was found in the K stock. Carbon stocks were highest for all plantations, followed by the K, N, and P stocks. Plantation soils exhibited a higher C:P ratio and a lower P:K ratio at various soil depths. The dominant controlling factors for the soil macronutrient stocks varied significantly at different stand ages and soil depths according to statistical analysis. For the total soil system, the C stock was affected by the available nutrients, organic matter, and stoichiometry; the available nutrients and organic matter were the determinant factors of the N and P stocks. Aggregate stability could be the primary parameter affecting the K stock. Organic matter explained most of the variation in soil macronutrient stocks, followed by the P:K ratio and available K. Collectively, our results suggest that the response of soil macronutrient stocks to stand age and soil depth will be dependent on different soil physicochemical properties, and P and K may be important limiting factors in Masson pine plantation ecosystems.

PmMYB4, a Transcriptional Activator from Pinus massoniana, Regulates Secondary Cell Wall Formation and Lignin Biosynthesis

Wood formation originates in the biosynthesis of lignin and further leads to secondary cell wall (SCW) biosynthesis in woody plants. Masson pine (Pinus massoniana Lamb) is an economically important industrial timber tree, and its wood yield affects the stable development of the paper industry. However, the regulatory mechanisms of SCW formation in Masson pine are still unclear. In this study, we characterized PmMYB4, which is a Pinus massoniana MYB gene involved in SCW biosynthesis. The open reading frame (ORF) of PmMYB4 was 1473 bp, which encoded a 490 aa protein and contained two distinctive R2 and R3 MYB domains. It was shown to be a transcription factor, with the highest expression in semi-lignified stems. We overexpressed PmMYB4 in tobacco. The results indicated that PmMYB4 overexpression increased lignin deposition, SCW thickness, and the expression of genes involved in SCW formation. Further analysis indicated that PmMYB4 bound to AC-box motifs and might directly activate the promoters of genes (PmPAL and PmCCoAOMT) involved in SCW biosynthesis. In addition, PmMYB4-OE(over expression) transgenic lines had higher lignin and cellulose contents and gene expression than control plants, indicating that PmMYB4 regulates SCW mainly by targeting lignin biosynthetic genes. In summary, this study illustrated the MYB-induced SCW mechanism in Masson pine and will facilitate enhanced lignin and cellulose synthesis in genetically engineered trees.

Management of Needle-Eating Caterpillars Associated with Pinus massoniana and P. merkusii in Vietnam

Pine tussock moth (Dasychira axutha Collenette (Lymantriidae)) and masson pine caterpillar (Dendrolimus punctatus Walker (Lasiocampidae)) cause serious damage to Pinus massoniana and P. merkusii plantations in Vietnam. An integrated pest management (IPM) program is required to control these pests. Therefore, this study was undertaken to identify damage thresholds and to evaluate control options for implementing IPM in the Northeast and North Central regions of Vietnam. Three damage thresholds were considered: <25%, 25–50% and >50% loss of leaf area. Control options were manual collection of eggs and pupae and the use of light traps and biological agents. An initial IPM model was developed for each pest and these gave control efficacies of 82.4% (Da. axutha on P. massoniana) and 77.8% (De. punctatus on P. merkusii). Six extended IPM models were undertaken by forest companies giving pest control efficacies of 79.2–85.9%. The collection of pine resin provides an important source of income to local communities and the resin yield in the IPM models increased by 20.9–22.7% compared to untreated control plots. The IPM protocols would help in developing policies that promote a more sustainable management of forest pests in Vietnam, thus reducing dependence on the use of insecticides of environmental and public health concerns.