Effects of Low-Temperature Stress on Physiological Characteristics and Microstructure of Stems and Leaves of Pinus massoniana L.

Pinus massoniana L. is one of the most important conifer species in southern China and is the mainstay of the forest ecosystem and timber production, yet low temperatures limit its growth and geographical distribution. This study used 30-day-old seedlings from families of varying cold-tolerance to examine the morphological traits of needles and stems, chlorophyll fluorescence characteristics, protective enzymes, and changes in starch and lignin under different low-temperature stresses in an artificial climate chamber. The results showed that the seedlings of Pinus massoniana exhibited changes in phenotypic morphology and tissue structure under low-temperature stress. Physiological and biochemical indexes such as protective enzymes, osmoregulatory substances, starch, and lignin responded to low-temperature stress. The cold-tolerant family increased soluble sugars, starch grain, and lignin content as well as peroxidase activity, and decreased malondialdehyde content by increasing the levels of actual photochemical efficiency (ΦPSII), electron transport rate (ETR), and photochemical quenching (qP) to improve the cold tolerance ability. This study provides a reference for the selection and breeding of cold-tolerant genetic resources of Pinus massoniana and the mechanism of cold-tolerance, as well as the analysis of the mechanism of adaptation of Pinus massoniana in different climatic regions of China.

Transcriptomic Identification of Potential C2H2 Zinc Finger Protein Transcription Factors in Pinus massoniana in Response to Biotic and Abiotic Stresses.

Biotic and abiotic stresses have already seriously restricted the growth and development of Pinus massoniana, thereby influencing the quality and yield of its wood and turpentine. Recent studies have shown that C2H2 zinc finger protein transcription factors play an important role in biotic and abiotic stress response. However, the members and expression patterns of C2H2 TFs in response to stresses in P. massoniana have not been performed. In this paper, 57 C2H2 zinc finger proteins of P. massoniana were identified and divided into five subgroups according to a phylogenetic analysis. In addition, six Q-type PmC2H2-ZFPs containing the plant-specific motif 'QALGGH' were selected for further study under different stresses. The findings demonstrated that PmC2H2-ZFPs exhibit responsiveness towards various abiotic stresses, including drought, NaCl, ABA, PEG, H2O2, etc., as well as biotic stress caused by the pine wood nematode. In addition, PmC2H2-4 and PmC2H2-20 were nuclear localization proteins, and PmC2H2-20 was a transcriptional activator. PmC2H2-20 was selected as a potential transcriptional regulator in response to various stresses in P. massoniana. These findings laid a foundation for further study on the role of PmC2H2-ZFPs in stress tolerance.

Effects of thinning and understory removal on soil phosphorus fractions in subtropical pine plantations.

Forest management changes the physical environments and nutrient dynamics and then regulates the forest productivity. Soil phosphorus (P) availability is critical for productivity in tropical and subtropical forests. However, it was still poorly understood how soil P content and fraction respond to various forest management practices in these regions. Here, we measured the soil total P, available P, and Hedley's P fractions, including inorganic and organic P (Pi and Po), in subtropical pine plantations treated with understory removal (UR), non-dominant species thinning (NDST) and dominant species thinning (DST) after nine years. Compared to plantations without management (CK), treatments such as UR, NDST, and DST decreased soil total P at 0-10 cm and soil available P at 0-10 cm and 10-20 cm. Increases in resin-Pi, NaOH-Pi, and C.HCl-Pi resulted in a higher total Pi in 0-10 cm (p < 0.05) in treated plots (UR, NDST, and DST) than in CK plots. UR, NDST, and DST treatments increased NaHCO3-Po and NaOH-Po (p < 0.05) but decreased C.HCl-Po at a depth of 0-10 cm. Regardless of management treatments, soil total P, available P, and P fractions in 0-10 cm showed higher contents than those in 10-20 cm. There were positive relationships between total P and total Po (p < 0.01) and between available P and total Pi. There were also positive relationships between total P, available P, NaHCO3-Pi, and NaOH-Pi (p < 0.05). In conclusion, forest management such as UR, NDST, and DST decreased soil total P and available P, and transforming soil P fractions to available P will meet the P demand following management in the pine plantations of subtropical China.

Estimation of changes in carbon sequestration and its economic value with various stand density and rotation age of Pinus massoniana plantations in China.

Plantations actively participate in the global carbon cycle and play a significant role in mitigating global climate change. However, the influence of forest management strategies, especially planting density management, on the biomass carbon storage and production value of plantations for ensuring carbon sink benefits is still unclear. In this study, we estimated the carbon sequestration and economic value of Pinus massoniana plantations with various stand densities and rotation ages using a growth model method. The results revealed that with increasing stand age, low-density plantations at 2000 trees·ha-1 (358.80 m3·ha-1), as well as high-density plantations at 4500 trees·ha-1 (359.10 m3·ha-1), exhibited nearly identical standing volumes, which indicated that reduced inter-tree competition intensity favors the growth of larger trees during later stages of development. Furthermore, an increase in planting density led to a decrease in the average carbon sequestration rate, carbon sink, and number of trees during the rapid growth period, indicating that broader spacing between trees is favorable for biomass carbon accumulation. Further, extending the rotation period from 15 to 20 years or 25 years and reducing the optimal planting density from 3000 to 2000 trees·ha-1 increased the overall benefits of combined timber and carbon sink income by 2.14 and 3.13 times, respectively. The results highlighted that optimizing the planting density positively impacts the timber productivity and carbon sink storage of Pinus massoniana plantations and boosts the expected profits of forest managers. Thus, future afforestation initiatives must consider stand age and planting density management to shift from a scale-speed pattern to a quality-benefit design.

Predicting the potential distribution of Dendrolimus punctatus and its host Pinus massoniana in China under climate change conditions.

Introduction: Dendrolimus punctatus, a major pest endemic to the native Pinus massoniana forests in China, displays major outbreak characteristics and causes severe destructiveness. In the context of global climate change, this study aims to investigate the effects of climatic variations on the distribution of D. punctatus and its host, P. massoniana. Methods: We predict their potential suitable distribution areas in the future, thereby offering a theoretical basis for monitoring and controlling D. punctatus, as well as conserving P. massoniana forest resources. By utilizing existing distribution data on D. punctatus and P. massoniana, coupled with relevant climatic variables, this study employs an optimized maximum entropy (MaxEnt) model for predictions. With feature combinations set as linear and product (LP) and the regularization multiplier at 0.1, the model strikes an optimal balance between complexity and accuracy. Results: The results indicate that the primary climatic factors influencing the distribution of D. punctatus and P. massoniana include the minimum temperature of the coldest month, annual temperature range, and annual precipitation. Under the influence of climate change, the distribution areas of P. massoniana and its pests exhibit a high degree of similarity, primarily concentrated in the region south of the Qinling-Huaihe line in China. In various climate scenarios, the suitable habitat areas for these two species may expand to varying degrees, exhibiting a tendency to shift toward higher latitude regions. Particularly under the high emission scenario (SSP5-8.5), D. punctatus is projected to expand northwards at the fastest rate. Discussion: By 2050, its migration direction is expected to closely align with that of P. massoniana, indicating that the pine forests will continue to be affected by the pest. These findings provide crucial empirical references for region-specific prevention of D. punctatus infestations and for the rational utilization and management of P. massoniana resources.

Community assembly of ectomycorrhizal fungal communities in pure and mixed Pinus massoniana forests.

Ectomycorrhizal (EcM) fungi play an important role in nutrient cycling and community ecological dynamics and are widely acknowledged as important components of forest ecosystems. However, little information is available regarding EcM fungal community structure or the possible relationship between EcM fungi, soil properties, and forestry activities in Pinus massoniana forests. In this study, we evaluated soil properties, extracellular enzyme activities, and fungal diversity and community composition in root and soil samples from pure Pinus massoniana natural forests, pure P. massoniana plantations, and P. massoniana and Liquidambar gracilipes mixed forests. The mixed forest showed the highest EcM fungal diversity in both root and bulk soil samples. Community composition and co-occurrence network structures differed significantly between forest types. Variation in the EcM fungal community was significantly correlated with the activities of ß-glucuronidase and ß-1,4-N-acetylglucosaminidase, whereas non-EcM fungal community characteristics were significantly correlated with ß-1,4-glucosidase and ß-glucuronidase activities. Furthermore, stochastic processes predominantly drove the assembly of both EcM and non-EcM fungal communities, while deterministic processes exerted greater influence on soil fungal communities in mixed forests compared to pure forests. Our findings may inform a deeper understanding of how the assembly processes and environmental roles of subterranean fungal communities differ between mixed and pure plantations and may provide insights for how to promote forest sustainability in subtropical areas.

The Growth-Promoting and Colonization of the Pine Endophytic Pseudomonas abietaniphila for Pine Wilt Disease Control.

In this study, we focused on evaluating the impact of Pseudomonas abietaniphila BHJ04 on the growth of Pinus massoniana seedlings and its biocontrol efficacy against pine wilt disease (PWD). Additionally, the colonization dynamics of P. abietaniphila BHJ04 on P. massoniana were examined. The growth promotion experiment showed that P. abietaniphila BHJ04 significantly promoted the growth of the branches and roots of P. massoniana. Pot control experiments indicated that strain BHJ04 significantly inhibited the spread of PWD. There were significant changes in the expression of several genes related to pine wood nematode defense in P. massoniana, including chitinase, nicotinamide synthetase, and triangular tetrapeptide-like superfamily protein isoform 9. Furthermore, our results revealed significant upregulation of genes associated with the water stress response (dehydration-responsive proteins), genetic material replication (DNA/RNA polymerase superfamily proteins), cell wall hydrolase, and detoxification (cytochrome P450 and cytochrome P450 monooxygenase superfamily genes) in the self-regulation of P. massoniana. Colonization experiments demonstrated that strain BHJ04 can colonize the roots, shoots, and leaves of P. massoniana, and the colonization amount on the leaves was the greatest, reaching 160,000 on the 15th day. However, colonization of the stems lasted longer, with the highest level of colonization observed after 45 d. This study provides a preliminary exploration of the growth-promoting and disease-preventing mechanisms of P. abietaniphila BHJ04 and its ability to colonize pines, thus providing a new biocontrol microbial resource for the biological control of plant diseases.

Comparative metabolomics and transcriptomics analyses provide insights into branching changes induced by heterografting in Pinus massoniana seedlings.

The investigation of dwarfing rootstocks for the establishment of high-generation seed orchards is a prospective avenue of research. In this investigation, Pinus massoniana, Pinus yunnanensis var. pygmaea (P. pygmaea), and P. elliottii seedlings were used as rootstocks for grafting with P. massoniana scions. Grafting P. massoniana onto P. pygmaea rootstock resulted in observable phenotypic alterations in lateral branches, apical buds, and needle length. Certain characteristic metabolites of rootstocks, such as fatty acyls, pregnenolones, steroids, and steroid derivatives, were found to be highly expressed in scions after grafting. RNA-seq analysis revealed MYB-related, SBP, and bHLH demonstrating a significant positive correlation, while C2H2 and Orphans exhibited negative correlations with the differential intensity of metabolites related to lipids and lipid-like molecules. This study offers valuable insights for the establishment of rootstock breeding programs.

Intraspecific transitioning of ecological strategies in Pinus massoniana trees across restoration stages.

Intraspecific variation in plant functional traits and ecological strategies is typically overlooked in most studies despite its pivotal role at the local scales and along short environmental gradients. While CSR theory has been used to classify ecological strategies (competitive C; stress-tolerant, S; ruderal, R) in different plant species, its ability to explain intraspecific variation in ecological strategies remains uncertain. Here, we sought to investigate intraspecific variation in ecological strategies for Pinus massoniana, a pioneer conifer tree for ecological restoration in Changting County, southeast China. By measuring key leaf traits and canopy height of 252 individuals at different ontogenetic stages from three plots spanning distinctive stages along early ecological restoration and calculating their C, S, and R scores, we constructed an intraspecific CSR system. All individual strategies shifted across three restoration stages, with adults from higher S component to higher C component while juveniles from higher S component to higher R component. Our results suggest that while strategies of all P. massoniana individuals start with tolerance to environmental stress, as restoration proceeds, adult transition towards completion for light, whereas juveniles shift to an acquisitive resource use. The study reveals an intraspecific pattern of strategy variation during forest restoration, contributing to our understanding of how plants adapt to diverse environments.

Functional Characterization of PmDXR, a Critical Rate-Limiting Enzyme, for Turpentine Biosynthesis in Masson Pine (Pinus massoniana Lamb.).

As one of the largest and most diverse classes of specialized metabolites in plants, terpenoids (oprenoid compounds, a type of bio-based material) are widely used in the fields of medicine and light chemical products. They are the most important secondary metabolites in coniferous species and play an important role in the defense system of conifers. Terpene synthesis can be promoted by regulating the expressions of terpene synthase genes, and the terpene biosynthesis pathway has basically been clarified in Pinus massoniana, in which there are multiple rate-limiting enzymes and the rate-limiting steps are difficult to determine, so the terpene synthase gene regulation mechanism has become a hot spot in research. Herein, we amplified a PmDXR gene (GenBank accession no. MK969119.1) of the MEP pathway (methyl-erythritol 4-phosphate) from Pinus massoniana. The DXR enzyme activity and chlorophyll a, chlorophyll b and carotenoid contents of overexpressed Arabidopsis showed positive regulation. The PmDXR gene promoter was a tissue-specific promoter and can respond to ABA, MeJA and GA stresses to drive the expression of the GUS reporter gene in N. benthamiana. The DXR enzyme was identified as a key rate-limiting enzyme in the MEP pathway and an effective target for terpene synthesis regulation in coniferous species, which can further lay the theoretical foundation for the molecularly assisted selection of high-yielding lipid germplasm of P. massoniana, as well as provide help in the pathogenesis of pine wood nematode disease.

Changes and driving factors of microbial community composition and functional groups during the decomposition of Pinus massoniana deadwood.

Clarifying changes in the microbial community in deadwood at different stages of decomposition is crucial for comprehending the role of deadwood in the biogeochemical processes and the sustainability of forest development. However, there have been no reports on the dynamics of microbial community during the decomposition of Pinus massoniana. We used the "space-for-time" substitution to analyze the characteristics of microbial community changes and the key influencing factors in the P. massoniana deadwood during different decomposition stages by 16S and ITS rRNA gene sequencing. The results suggest that the microbial community structure of the early decomposition (decay class I) was significantly different from the other decay classes, while the diversity and richness of the microbial community were the highest in the late decomposition (decay class V). The Linear Discriminant Analysis Effect Size analysis revealed that most bacterial and fungal taxa were significantly enriched in decay classes I and V deadwood. During the initial stages of decomposition, the relative abundance of the bacterial functional group responsible for carbohydrate metabolism was greater than the later stages. As decomposition progressed, the relative abundance of saprophytic fungi gradually decreased, and there was a shift in the comparative abundance of mixed saprophytic-symbiotic fungi from low to high before eventually decreasing. Total organic carbon, total nitrogen, carbon-to-nitrogen ratio, total potassium, total phenol, condensed tannin, lignin, and cellulose were significantly correlated with microbial community structure, with the carbon-to-nitrogen ratio having the greatest effect. Our results indicate that the physicochemical properties of deadwood, microbial community structural composition and functional group changes were related to the decay class, among which the carbon-to-nitrogen ratio may be an important factor affecting the composition and diversity of microbial communities.

Comparative analysis of differential gene expression reveals novel insights into the heteroblastic foliage functional traits of Pinus massoniana seedlings.

Pinus massoniana needles, rich in medicinal polysaccharides and flavonoids, undergo heteroblastic foliage, transitioning from primary needles (PN) to secondary needles (SN) during growth, resulting in altered functional traits. Despite its significance, the molecular regulatory mechanisms governing these traits remain unclear. This study employs Iso-Seq and RNA-Seq analyses to explore differentially expressed genes (DEGs) associated with functional traits throughout the main growth season of heteroblastic foliage. Co-expression network analysis identified 34 hub genes and 17 key transcription factors (TFs) influencing light-harvesting antenna, photosystem I and II, crucial in photosynthesis regulation. Additionally, 14 genes involved in polysaccharide metabolism pathways, synthesizing sucrose, glucose, UDP sugars, and xylan, along with four genes in flavonoid biosynthesis pathways, regulating p-coumaroyl-CoA, quercetin, galangin, and myricetin production, exhibited differential expression between PN and SN. Further analysis unveils a highly interconnected network among these genes, forming a pivotal cascade of TFs and DEGs. Therefore, heteroblastic changes significantly impact needle functional traits, potentially affecting the pharmacological properties of PN and SN. Thus, these genomic insights into understanding the molecular-level differences of heteroblastic foliage, thereby establishing a foundation for advancements in the pharmaceutical industry related to needle-derived products.

Analysis on metabolic functions of rhizosphere microbial communities of Pinus massoniana provenances with different carbon storage by Biolog Eco microplates.

Introduction: Rhizosphere microorganisms are influenced by vegetation. Meanwhile, they respond to vegetation through their own changes, developing an interactive feedback system between microorganisms and vegetation. However, it is still unclear whether the functional diversity of rhizosphere soil microorganisms varies with different carbon storage levels and what factors affect the functional diversity of rhizosphere soil microorganisms. Methods: In this study, the Biolog-Eco microplate technique was used to analyze the metabolic diversity of carbon source of rhizosphere soil microorganisms from 6 Pinus massoniana provenances with three levels of high, medium and low carbon storage. Results: The results showed that the average well color development(AWCD) value of rhizosphere microorganisms was significantly positive correlated with carbon storage level of Pinus massoniana (p < 0.05). The AWCD value, Simpson and Shannon diversity of high carbon sequestrance provenances were 1.40 (144h incubation) 0.96 and 3.24, respectively, which were significantly higher (p < 0.05) than those of other P. massoniana provenances. The rhizosphere microbial AWCD, Shannon and Simpson diversity of the 6 provenances showed the same variation trend (SM>AY>QJ>SX>HF>SW). Similarly, microbial biomass carbon (MBC) content was positively correlated with carbon storage level, and there were significant differences among high, medium and low carbon storage provenances. The PCA results showed that the differences in the carbon source metabolism of rhizosphere microorganisms were mainly reflected in the utilization of amino acids, carboxylic acids and carbohydrates. Pearson correlation analysis showed that soil organic carbon (SOC), total nitrogen (TN) and pH were significantly correlated with rhizosphere AWCD (p < 0.05). Conclusion: Soil properties are important factors affecting rhizosphere microbial carbon source metabolism. The study confirmed that the microorganisms of high carbon storage provenances had relatively high carbon metabolic activity. Among them, the carbon metabolic activity of rhizosphere microorganisms of SM provenance was the highest, which was the preferred provenances in effective ecological service function.

[Influencing mechanism of stand age to the accumulation of microbial residue carbon in the Pinus masso-niana plantations]. / 林龄对马尾松人工林微生物残体碳积累的影响机制.

Clarifying the accumulation pattern of soil microbial residue carbon and its contribution to soil organic carbon (SOC) across stand age is helpful to understand the mechanism underlying soil carbon cycling. In this study, we analyzed the differences of amino sugar content, physicochemical properties and microbial composition in surface soil (0-10 cm) in young (6 a), middle-aged (13 a), near-mature (29 a), mature (38 a) and over-mature (57 a) Pinus massoniana plantations of subtropical China, quantified the microbial residue carbon content and its contribution to SOC, and discussed the mechanism. The results showed that SOC, total nitrogen, amorphous iron oxide and leucine aminopeptidase contents in the middle-aged plantation were significantly lower than those in the mature plantation. Soil pH and fungal/bacteria in young plantation were significantly higher than those in other age groups. Across the stand age gradient, the ranges of microbial, fungal and bacterial residue carbon were 7.52-14.63, 4.03-8.00 and 3.48-6.63 g·kg-1, respectively. The contents of all the residue carbon were significantly higher in the mature plantation than that of the middle-aged plantation, which were positively affected by soil total nitrogen content. The contribution of microbial, fungal, and bacterial residue carbon to SOC was 59.7%-72.3%, 33.4%-45.6%, and 24.3%-30.8%, respectively. The contribution of fungal residue carbon to SOC in young plantation was significantly higher than that in other age groups, and the contribution of bacterial residue carbon to SOC in middle-aged plantation was significantly higher than that in young and near-mature plantations, both of which were affected by soil inorganic nitrogen. Fungal residue carbon content was 1.2-1.7 times as that of bacterial residue carbon content, and dominated for the accumulation of microbial residue carbon. Results of the partial least squares model showed that stand age, soil environmental factors (such as leucine aminopeptidase, amorphous iron oxide, pH, and total nitrogen), bacterial residue carbon, fungal residue carbon and the contribution of bacterial residue carbon to SOC had total effects on the contribution of fungal residue carbon to SOC (-0.37, -1.16, 0.90, 1.09, and 0.83, respectively). In conclusion, stand age promoted the accumulation of microbial residue carbon but did not increase its contribution to SOC.

[Effects of mixed broadleaved tree species with pure Pinus massoniana plantation on soil microbial necromass carbon and organic carbon fractions]. / é©¬å°¾æ¾çº¯æž—æ··äº¤æ”¹é€ å¯¹åœŸå£¤å¾®ç”Ÿç‰©æ®‹ä½“ç¢³å’Œæœ‰æœºç¢³ç»„åˆ†çš„å½±å“.

Mixing native broadleaved tree species is a widely used method for renovating Pinus massoniana plantations. Soil microbial necromass carbon and organic carbon fractions are important parameters for evaluating the impacts of tree species mixing and soil organic carbon (SOC) stability. However, their responses to the mixing and renovation of P. massoniana plantation has not been understood yet. Here, we selected a pure P. massoniana plantation (PP) and a mixed P. massoniana and Castanopsis hystrix plantation, with ages of 16 (MP16) and 38 years (MP38), respectively, as the research objects. We quantified soil physical and chemical properties, microbial necromass carbon content, and organic carbon components at different soil layers to reveal whether and how the introduction of C. hystrix into P. massoniana plantation affected soil microbial necromass carbon and organic carbon components. The results showed that the mixed P. massoniana and C. hystrix plantation significantly reduced fungal necromass carbon content and the ratio of fungal/bacterial necromass carbon in the 0-20 cm and 20-40 cm soil layers. There were no significant differences in microbial necromass carbon contents, bacterial necromass carbon contents, and their contributions to SOC among the different plantations. The contribution of fungal necromass carbon to SOC was higher than that of bacterial necromass carbon in all plantation types. The contribution of soil mineral-associated organic carbon (MAOC) to SOC was higher than that of occluded particulate organic carbon (oPOC) and light-free particulate organic carbon (fPOC) for all plantation types. Mixing the precious broadleaved tree species (i.e., C. hystrix) with coniferous species (P. massoniana) significantly increased MAOC content and the contribution of MAOC, oPOC, and fPOC to SOC in the 0-20 cm and 20-40 cm soil layers. The MAOC of MP38 was significantly higher than that of PP in all soil layers and the MAOC of MP38 stands were significantly higher than MP16 stands in the 20-40 cm, 40-60 cm, and 60-100 cm soil layers, indicating that hybridization enhanced SOC stability and that the SOC of MP38 stands were more stable than MP16 stands. SOC and total nitrogen contents were the main environmental factors driving the changes in soil microbial necromass carbon, while soil total nitrogen and organically complexed Fe-Al oxides were the primary factors affecting organic carbon fraction. Therefore, SOC stability can be enhanced by introducing native broadleaved species, such as C. hystrix, during the management of the P. massoniana plantation.

A Bursaphelenchus xylophilus effector BxICD1 inducing plant cell death, concurrently contributes to nematode virulence and migration.

The migratory endoparasitic phytonematodes Bursaphelenchus xylophilus is the causal agent of pine wilt disease and causes significant economic damage to pine forests in China. Effectors play a key role in the successful parasitism of plants by phytonematodes. In this study, 210 genes obtained by transcriptomics analyses were found to be upregulated in B. xylophilus infecting Pinus massoniana that were not functionally annotated nor reported previously in B. xylophilus infecting P. thunbergii. Among these differentially expressed genes, a novel effector, BxICD1, that could induce cell death in the extracellular space of Nicotiana benthamiana was identified. BxICD1 was upregulated in the early stages of infection, as shown by RT-qPCR analyses. In situ hybridization analysis showed that BxICD1 was expressed in the esophageal gland of nematodes. The yeast signal sequence trap system indicated that BxICD1 possessed an N-terminal signal peptide with secretion functionality. Using an Agrobacterium-mediated transient expression system, it was demonstrated that the cell death-inducing activity of BxICD1 was dependent on N. benthamiana brassinosteroid-insensitive 1-associated kinase 1 (NbBAK1). Finally, BxICD1 contributed to B. xylophilus virulence and migration in host pine trees, as demonstrated by RNAi silencing assays. These findings indicate that BxICD1 both induces plant cell death and also contributes to nematode virulence and migration in P. massonian.

The influence of stand composition and season on canopy structure and understory light environment in different subtropical montane Pinus massoniana forests.

Canopy structure and understory light have important effects on forest productivity and the growth and distribution of the understory. However, the effects of stand composition and season on canopy structure and understory light environment (ULE) in the subtropical mountain Pinus massoniana forest system are poorly understood. In this study, the natural secondary P. massoniana-Castanopsis eyrei mixed forest (MF) and P. massoniana plantation forest (PF) were investigated. The study utilized Gap Light Analyzer 2.0 software to process photographs, extracting two key canopy parameters, canopy openness (CO) and leaf area index (LAI). Additionally, data on the transmitted direct (Tdir), diffuse (Tdif), and total (Ttot) radiation in the light environment were obtained. Seasonal variations in canopy structure, the ULE, and spatial heterogeneity were analyzed in the two P. massoniana forest stands. The results showed highly significant (P < 0.01) differences in canopy structure and ULE indices among different P. massoniana forest types and seasons. CO and ULE indices (Tdir, Tdif, and Ttot) were significantly lower in the MF than in the PF, while LAI was notably higher in the MF than in the PF. CO was lower in summer than in winter, and both LAI and ULE indices were markedly higher in summer than in winter. In addition, canopy structure and ULE indices varied significantly among different types of P. massoniana stands. The LAI heterogeneity was lower in the MF than in the PF, and Tdir heterogeneity was higher in summer than in winter. Meanwhile, canopy structure and ULE indices were predominantly influenced by structural factors, with spatial correlations at the 10 m scale. Our results revealed that forest type and season were important factors affecting canopy structure, ULE characteristics, and heterogeneity of P. massoniana forests in subtropical mountains.

Identification and Expression Patterns of WOX Transcription Factors under Abiotic Stresses in Pinus massoniana.

WUSCHEL-related homeobox (WOX) transcription factors (TFs) play a crucial role in regulating plant development and responding to various abiotic stresses. However, the members and functions of WOX proteins in Pinus massoniana remain unclear. In this study, a total of 11 WOX genes were identified, and bioinformatics methods were used for preliminary identification and analysis. The phylogenetic tree revealed that most PmWOXs were distributed in ancient and WUS clades, with only one member found in the intermediate clade. We selected four highly conserved WOX genes within plants for further expression analysis. These genes exhibited expressions across almost all tissues, while PmWOX2, PmWOX3, and PmWOX4 showed high expression levels in the callus, suggesting their potential involvement in specific functions during callus development. Expression patterns under different abiotic stresses indicated that PmWOXs could participate in resisting multiple stresses in P. massoniana. The identification and preliminary analysis of PmWOXs lay the foundation for further research on analyzing the resistance molecular mechanism of P. massoniana to abiotic stresses.

Response mechanism of carbon metabolism of Pinus massoniana to gradient high temperature and drought stress.

BACKGROUND: The carbon metabolism pathway is of paramount importance for the growth and development of plants, exerting a pivotal regulatory role in stress responses. The exacerbation of drought impacts on the plant carbon cycle due to global warming necessitates comprehensive investigation into the response mechanisms of Masson Pine (Pinus massoniana Lamb.), an exemplary pioneer drought-tolerant tree, thereby establishing a foundation for predicting future forest ecosystem responses to climate change. RESULTS: The seedlings of Masson Pine were utilized as experimental materials in this study, and the transcriptome, metabolome, and photosynthesis were assessed under varying temperatures and drought intensities. The findings demonstrated that the impact of high temperature and drought on the photosynthetic rate and transpiration rate of Masson Pine seedlings was more pronounced compared to individual stressors. The analysis of transcriptome data revealed that the carbon metabolic pathways of Masson Pine seedlings were significantly influenced by high temperature and drought co-stress, with a particular impact on genes involved in starch and sucrose metabolism. The metabolome analysis revealed that only trehalose and Galactose 1-phosphate were specifically associated with the starch and sucrose metabolic pathways. Furthermore, the trehalose metabolic heat map was constructed by integrating metabolome and transcriptome data, revealing a significant increase in trehalose levels across all three comparison groups. Additionally, the PmTPS1, PmTPS5, and PmTPPD genes were identified as key regulatory genes governing trehalose accumulation. CONCLUSIONS: The combined effects of high temperature and drought on photosynthetic rate, transpiration rate, transcriptome, and metabolome were more pronounced than those induced by either high temperature or drought alone. Starch and sucrose metabolism emerged as the pivotal carbon metabolic pathways in response to high temperature and drought stress in Masson pine. Trehalose along with PmTPS1, PmTPS5, and PmTPPD genes played crucial roles as metabolites and key regulators within the starch and sucrose metabolism.

Physiological and Transcriptome Responses of Pinus massoniana Seedlings Inoculated by Various Ecotypes of the Ectomycorrhizal Fungus Cenococcum geophilum during the Early Stage of Drought Stress.

The impact of drought stress on plant growth in arid regions is a critical concern, necessitating the exploration of strategies to enhance plant drought resistance, particularly during the early stages of drought stress. This study focuses on the ectomycorrhizal fungus Cenococcum geophilum, renowned for its extensive genetic diversity and broad host compatibility, making it a crucial ally for host plants facing external stresses. We utilized Pinus massoniana seedlings inoculated with different ecotypic strains of C. geophilum under drought stress. The results showed that the inoculation of most strains of C. geophilum enhanced the drought resistance of P. massoniana seedlings under the early stages of drought stress, by influencing the water content, photosynthesis, accumulation of osmotic adjustment substances, and antioxidant enzyme activities in both shoots and roots of seedlings. Transcriptome analysis showed that mycorrhizal seedlings mainly regulated energy metabolism and reduction-oxidation reaction to resist early drought stress. Notably, the level of drought resistance observed in mycorrhizal seedlings was irrespective of the level of drought tolerance of C. geophilum strains. This study contributes essential data for understanding the drought response mechanisms of mycorrhizal P. massoniana seedlings inoculated by distinct C. geophilum ecotypes and guidance on selecting candidate species of ectomycorrhizal fungi for mycorrhizal afforestation in drought areas.