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Drought stress poses a serious threat to Pinus massoniana seedling growth in southern China. Trichoderma species, as beneficial microorganisms, have been widely used in agriculture to enhance plant growth and drought tolerance, but the interaction mechanisms remain unclear. To investigate the effect of drought-resistant Trichoderma longibrachiatum inoculation on P. massoniana growth under drought stress, the plant physiological indicators and rhizosphere microbiome diversity were measured to identify Trichoderma-activated mechanisms. Trichoderma longibrachiatum inoculation significantly promoted P. massoniana growth under drought treatment, and enhanced nitrogen, phosphorus, and potassium absorption compared with those of non-inoculated seedlings. Trichoderma longibrachiatum treatment alleviated the damage to cell membranes and needle tissue structure, and significantly increased antioxidant enzyme activities, osmotic substance contents, and photosynthesis in P. massoniana in response to drought stress. Soil nutrient contents, activities of sucrase, phosphatase, and urease as well as the relative abundances of the dominant genera Burkholderia, Rhodanobacter, and Trichoderma were elevated in the rhizosphere soil of P. massoniana inoculated with T. longibrachiatum under drought stress. A network analysis showed that certain crucial dominant taxa driven by T. longibrachiatum inoculation, including Penicillium, Trichoderma, Simplicillium, Saitozyma, Burkholderia, Bradyrhizobium, Sinomonas, and Mycobacterium, had more correlations with other microorganisms in the soil. Trichoderma longibrachiatum enhanced P. massoniana seedling growth under drought stress by regulating physiological responses and soil microbial community. [ABSTRACT FROM AUTHOR]

Studies on the dynamics of non-structural carbohydrates (NSCs) play an important role in understanding the mechanisms of plant responses to drought stress. The objective of this study was to assess the influence of ectomycorrhizal fungi (ECMF) on the content and distribution of NSCs in Pinus massoniana seedlings under different drought intensities and to further explore the possible mechanism by which ECMF enhances the stress resistance of host plants. We conducted a pot experiment using P. massoniana seedlings that were inoculated (M) or non-inoculated (NM) with Suillus luteus (Sl) under well-watered, moderate, and severe drought stress conditions. The results showed that drought significantly reduced the photosynthetic capacity of P. massoniana seedlings and inhibited their growth rate. P. massoniana could respond to different degrees of drought stress by increasing the accumulation of NSCs and increasing WUE. However, compared with well-watered treatment, NSCs consumption began to appear in the roots of NM due to the decrease in starch content under severe drought, whereas NSCs content in M seedlings was higher than that in the well-watered treatment, showing that the ability to maintain C balance was higher in M seedlings. Compared with NM, inoculation with Sl increased the growth rate and biomass of roots, stems, and leaves under moderate and severe drought. In addition, Sl can also improve the gas exchange parameters (net photosynthetic rate, transpiration rate, intercellular CO2 concentration and stomatal conductance) of P. massoniana seedlings compared with NM seedlings, which was conducive to the hydraulic regulation of seedlings and improved their C fixation capacity. Meanwhile, the content of NSCs in M seedlings was higher. Moreover, the soluble sugar content and SS/St ratio of leaves, roots, and whole plants were higher under drought stress after Sl inoculation, indicating that Sl could also change the C distribution mode, regulate more soluble sugar to respond to drought stress, which was conducive to improving the osmotic adjustment ability of seedlings, and providing more available C sources for plant growth and defense. Overall, inoculation with Sl could enhance the drought resistance of seedlings and promote their growth under drought stress by improving NSCs storage, increasing soluble sugar distribution, and improving the plant water balance of P. massoniana seedlings. [ABSTRACT FROM AUTHOR]

Pruning is an important technique in culturing good knot-free timber. However, to make more accurate pruning plans, it is necessary to consider the growing status of trees and set reasonable pruning intensities based on this. In a seven-year-old Pinus massoniana Lamb. plantation, we carried out pruning twice with a time interval of two years. The treatments included one unpruned treatment (CK) and five pruned treatments (from P1 to P5, representing the live branch height being kept at 68%, 55%, 60%, 55% and 45% of the tree height). CK, P1 and P2 were conducted in the first pruning in March 2019, and the remaining treatments were conducted in the second pruning in January 2021. The growth investigations were carried out in March 2019, December 2019, December 2020 and December 2021. Another investigation was carried out only for the measurement of live branch heights and crown widths in January 2021 just after the second pruning. The results showed that pruning resulted in a 15.08% to 60.62% increase in diameter growth and an 10.28% to 29.87% increase in volume growth. The stem form was also improved. Significant differences were recorded in live branch heights after green pruning but gradually recovered to the same level in two years by branch senescence. Pruning also resulted in a faster extension of the crown width with an enlarged growing space by the removal of green branches. We highlighted that trees with different growth statuses responded differently to pruning intensities: trees of weak growth statuses grew better under a light pruning intensity, while those with a strong growth status grew better under a severe pruning intensity. Overall, for the seven-year-old young mason pine plantation, keeping four rounds of branches in trees with diameters of less than 8 cm and keeping three rounds of branches in trees with diameters of more than 8 cm were appropriate measures. [ABSTRACT FROM AUTHOR]

Nutrient resorption and stoichiometry ratios are vital indicators to explore nutrient transfer and use efficiency for plants, particularly under the condition of nutrient limitation. However, the changing rules about nutrient resorption and ecological stoichiometry homeostasis are still unclear with the development of plantations. We determined carbon (C), nitrogen (N), and phosphorus (P) concentrations in soil and in fresh and senesced needles along a chronosequence of Pinus massoniana plantations (10, 20, 30, and 36 years old) in Guizhou Province, China. We also calculated the N and P resorption efficiency (NRE and PRE, respectively) and the homeostasis coefficient. The results showed that fresh and senesced needles' C and N concentrations maintained an increasing trend, whereas their P concentrations decreased initially and subsequently increased as the plantations' ages increased. Fresh needles' N:P ratios indicated that N limitation existed before 20 years old, while P limitation appeared in the 30-year-old plantations. The NRE and PRE showed patterns of increasing initially and decreasing subsequently along the chronosequence of P. massoniana plantations, which was coupled with weak stoichiometric homeostasis to reduce nutrient deficiency. Therefore, the appropriate nutrient management measurements should be induced to promote tree growth and the sustainable development of P. massoniana plantations. [ABSTRACT FROM AUTHOR]

Ectomycorrhiza (ECM) function has been well studied; however, there is little detailed information regarding the establishment of ECM symbioses. We investigated the morphological and transcriptional changes that occur during the establishment of the Pinus massoniana–Suillus bovinus ECM. S. bovinus promoted the growth of P. massoniana via the release of volatile organic compounds and exudates during the pre-symbiotic stage. Exudate-induced effects showed host plant specificity. At seven days post-inoculation (dpi), the mycelium started to penetrate P. massoniana roots. At 28 dpi, the Hartig net and mantle formed. At the pre-symbiotic stage, most differentially expressed genes in P. massoniana roots were mapped to the biosynthesis of secondary metabolites, signal transduction, and carbohydrate metabolism. At the symbiotic stage, S. bovinus colonization induced the reprogramming of pathways involved in genetic information processing in P. massoniana, particularly at the Hartig net and mantle formation stage. Phenylpropanoid biosynthesis was present at all stages and was regulated via S. bovinus colonization. Enzyme inhibitor tests suggested that hydroxycinnamoyl-CoA shikimate/quinate transferase is involved in the development of the Hartig net. Our findings outline the mechanism involved in the P. massoniana–S. bovinus ECM. Further studies are needed to clarify the role of phenylpropanoid biosynthesis in ECM formation. [ABSTRACT FROM AUTHOR]

The spatial heterogeneity on a regional scale of forest biomass is caused by multiple biotic and abiotic factors. However, the contributions of biotic and abiotic factors to the spatial heterogeneity of forest biomass remain unclear. Based on the data of the National Forest Continuous Inventory (NFCI), digital elevation model (DEM), and meteorological data of Guizhou Province in 2015, we studied the spatial heterogeneity of the aboveground forest biomass in Guizhou province and evaluated the contribution rates of its influencing factors using Moran's I, semivariogram, distance-based Moran's eigenvector maps (dbMEMs), and variance partitioning. The results showed that the forest biomass in Guizhou province had strong spatial heterogeneity. Biotic and abiotic factors explained 34.4% and 19.2% of the spatial variation in forest biomass, respectively. Among the biotic factors, the average height of the stand had the greatest influence on forest biomass, while annual precipitation had the greatest influence on forest biomass among abiotic factors. Spatial factors only explained 0.7% of the spatial variation of forest biomass, indicating that the contribution of spatial factors can be explained by some measured abiotic factors. This study provided an effective approach to understand the underlying mechanisms of spatial allocation of forest biomass. [ABSTRACT FROM AUTHOR]

In recent years, global forests have been facing an increase in tree mortality owing to increasing droughts. However, the capacity for plants to adjust their physiology and biochemistry during extreme drought and subsequent recovery is still unclear. Here, we used 1.5-year-old Pinus massoniana Lamb. seedlings and simulated drought conditions to achieve three target stress levels (50%, 85%, and 100% loss of stem hydraulic conductivity (PLC)), followed by rehydration. Needle water status, gas exchange, and biochemical parameters were assessed during drought and recovery. The results showed that drought had significantly negative impacts on needle water status and gas exchange parameters, with gas exchange declining to 0 after PLC85 was achieved. Soluble protein concentration (SPC), soluble sugar concentration (SSC), malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, and needle water-use efficiency showed fluctuations. The activity of antioxidant enzymes and the values of osmotic regulators were then gradually decreased as the physiological and biochemical functions of seedlings were disturbed. Seedlings showed a stronger ability to recover from PLC50 than PLC85 and PLC100. We conclude that the physiological and biochemical recovery of P. massoniana seedlings is more likely to be inhibited when plants experience increasing drought stress that induces 85% and greater loss of hydraulic conductance. [ABSTRACT FROM AUTHOR]

Pines have heteroblastic foliage (primary and secondary needles) during seedling stage, but how heteroblastic foliage affects carbon storage and biomass accumulation, contributing to seedling quality, is unclear. We investigated the influences of heteroblastic foliage on photosynthetic physiological characteristics, non-structural carbohydrate (NSC) and biomass accumulation in current-year seedlings; the key factors determining biomass accumulation were mainly determined by principal component screening, Spearman correlation, and path analysis. The results indicated that (1) primary needles have high photosynthetic pigments (chlorophyll a and total chlorophyll), net photosynthetic rates (Pn), the potential maximum photochemical efficiency (Fv/Fm), and leaf instantaneous water use efficiency (WUEi), whereas higher non-photochemical quenching (NPQ) suggested that sudden light increases induce the initiation of quenching mechanism in primary needles; additionally, secondary needles had a lower transpiration rate (Tr), limiting stomata (Ls), and light saturation point. (2) Secondary needles promoted soluble sugar (fructose and glucose) increases in leaves compared to that of primary needles and increased the leaf biomass accumulation (from 47.06% to 54.30%), enhancing the overall ability of photosynthetic organs; additionally, secondary needles can enhance the proportion of starch storage in the roots, and NSC accumulation was significantly increasing in the seedling leaves and roots. (3) Photosynthetic pigments (carotenoids, chlorophyll a, and total chlorophyll) had direct positive effects on primary needle seedling (PNS) biomass and promoted biomass by indirectly increasing soluble sugar synthesis in the stems. The Pn was the main physiological factor determining PNS biomass accumulation. In addition, the WUEi, Ls, and NPQ had direct negative effects on PNS biomass accumulation, inhibiting photosynthesis to limit seedling growth. Considering the functional traits in heteroblastic foliage is necessary when assessing different leaf types of Pinus massoniana (Lamb.) seedlings, in particular those threats implicated in light, water, and temperature relations. Our results can be beneficial to guide the establishment of seedling management and afforestation measures. [ABSTRACT FROM AUTHOR]

With increasing age, plants will cause changes in soil physicochemical properties. The objective of this study was to investigate differences in the soil physicochemical properties in different-aged Masson pine forest plantations (i.e., 10, 20, 40, and 60 years old). Soil samples were collected in a pure Masson pine forest plantation in Southwest China. The soil determination indexes included organic carbon, nitrogen, phosphorus and potassium contents, water content, bulk density, and pH. The soil pH of a 20-year-old forest was significantly (p < 0.05) higher than that of a 10, 40, and 60-year-old forest. In addition, soil-available phosphorus in a 60-year-old forest was significantly (p < 0.05) higher than that in the other three age forest groups. With increasing forest age, available phosphorus increased, while available nitrogen decreased at 20 years old and then increased at 40 years old. There was a significant positive correlation (p < 0.05) between total nitrogen and available potassium; no significant correlation (p> 0.05) between total phosphorus and total potassium, organic carbon, bulk density, and pH; and a significant negative correlation (p < 0.05) between available phosphorus and the water content. The availability and utilization efficiency of soil nutrients in young forests were higher than those in old forests and the intermediate forest age was an important time point that affected the soil properties. To improve the availability of soil nutrients and ensure the sustainable utilization of soil resources, it is necessary to increase the input of nitrogen and especially phosphorus. More attention should be given to the phytochemometric response with respect to the age of plantations. [ABSTRACT FROM AUTHOR]

Brassinosteroids (BRs) are known to be essential regulators for wood formation in herbaceous plants and poplar, but their roles in secondary growth and xylem development are still not well-defined, especially in pines. Here, we treated Pinus massoniana seedlings with different concentrations of exogenous BRs, and assayed the effects on plant growth, xylem development, endogenous phytohormone contents and gene expression within stems. Application of exogenous BR resulted in improving development of xylem more than phloem, and promoting xylem development in a dosage-dependent manner in a certain concentration rage. Endogenous hormone determination showed that BR may interact with other phytohormones in regulating xylem development. RNA-seq analysis revealed that some conventional phenylpropanoid biosynthesis- or lignin synthesis-related genes were downregulated, but the lignin content was elevated, suggesting that new lignin synthesis pathways or other cell wall components should be activated by BR treatment in P. massoniana. The results presented here reveal the foundational role of BRs in regulating plant secondary growth, and provide the basis for understanding molecular mechanisms of xylem development in P. massoniana. [ABSTRACT FROM AUTHOR]

Flavonoids in Rosa sterilis were studied. The flavonoids in Rosa sterilis were extracted by ultrasonic method, and the extraction conditions were modeled and optimized by response the surface methodology and the artificial intelligence method. The results show that the ultrasonic method can effectively extract total flavonoids, and the extraction rate is close to the prediction value of ANN-GA algorithm, which proves the rationality of the model. The order of the effects of the parameters on the experiment was material liquid ratio > extraction power > extraction time > ethanol concentration. In addition, the scavenging effects of flavonoids on DPPH, O2−· and ·OH were also determined, and these indicated that flavonoids have strong antioxidant activities. The kinetics of the extraction process was studied by using the data of the extraction process, and it was found that the extraction process conformed to Fick's first law. [ABSTRACT FROM AUTHOR]

Light intensity is a critical factor regulating photosynthetic capacity in plants. However, the effects of varying light intensity on morphological and photoprotective mechanisms in Pinus massoniana seedlings have not been explored in depth, especially those in the first seedling growing season. We measured the growth, photosynthetic physiology, biochemistry, and chlorophyll fluorescence of P. massoniana seedlings at four light gradients: 100% relative irradiance (RI, full sunlight), 70% RI, 50% RI, and 20% RI. The seedling height at 70% RI was 9.27% higher than that at 100% RI. However, seedling height was inhibited under low light intensity; at 20% RI, all seedlings died. The decreasing light intensity inhibited ground diameter growth but increased the height-diameter ratio. The secondary needle emergence rate was 53.4% higher at 70% RI than at 100% RI but was only 2% at 50% RI. The chlorophyll and carotenoid contents increased significantly with decreasing light intensity. The increased Chl b and Car contents promoted the photoreceptor potential of the violet (400~420 nm), blue (440~480 nm), and yellow-orange (597~655 nm) regions in leaves. Among the chlorophyll fluorescence parameters, Fv/Fm, Fv′/Fm′, Y(II), qp, and ETR all reached maximum values at 70% RI but were significantly lower at 50% RI than at 100% RI. However, decreasing the light intensity caused a reduction in NPQ. The 70% RI level increased POD and SOD activity and the contents of osmotic regulation substances and slowed MDA accumulation. Seedlings at 70% RI had a higher growth rate, higher photosynthetic activity and potential, and significantly greater stress resistance than the other seedlings. Therefore, appropriate shading measures were beneficial to the cultivation of vigorous seedlings. Furthermore, spectral reflectance indexes were found to be a suitable tool for monitoring the photosynthetic physiological characteristics, stress resistance characteristics, and growth status of P. massoniana seedlings in real time. [ABSTRACT FROM AUTHOR]

Masson pine is an important afforestation species in southern China, where seasonal drought is common. The present study focused on the effects of Suillus placidus, an ectomycorrhizal fungus, inoculation on the growth and physiological and biochemical performance of masson pine seedlings under four different watering treatments (well-watered, mild drought, moderate drought, and severe drought) to evaluate the symbiotic relationship between S. placidus and masson pine seedlings. Ectomycorrhizal-inoculated (ECM) and non-inoculated (NM) seedlings were grown in pots and maintained for 60 days using the weighing method. Results showed that seedlings' growth, dry weight, RWC, chlorophyll content, PSII efficiency, and photosynthesis decreased as drought stress intensified in both ECM and NM plants. This suggests that drought stress significantly limits the growth and photosynthetic performance of masson pine seedlings. Nevertheless, increased An/gs and proline contents in both NM and ECM prevented oxidative damage caused by drought stress. In addition, increased peroxidase (POD) activity is an essential defense mechanism of ECM seedling under drought stress. Compared with NM, ECM seedlings showed faster growth, higher RWC, and photosynthetic performance, and lower lipid peroxidation in cell membranes under drought stress, as indicated by higher POD activity and lower proline and malondialdehyde (MDA). Our experiment found that S. placidus inoculation can enhance the drought resistance of masson pine seedlings by increasing antioxidant enzyme activity, water use efficiency, and proline content, thereby enhancing growth under water-deficiency conditions. S. placidus can be used to cultivate high-quality seedlings and improve their survival in regions that experience seasonal droughts. [ABSTRACT FROM AUTHOR]

Global warming significantly impacts forest range areas by increasing soil acidification or aluminum toxicity. Aluminum (Al) toxicity retards plant growth by inhibiting the root development process, hindering water uptake, and limiting the bioavailability of other essential micronutrients. Pinus massoniana (masson pine), globally recognized as a reforestation plant, is resistant to stress conditions including biotic and abiotic stresses. This resistance is linked to the symbiotic relationship with diverse ectomycorrhizal fungal species. In the present study, we investigated the genetic regulators as expressed proteins, conferring a symbiotic relationship between Al-stress resistance and Suillus luteus in masson pine. Multi-treatment trials resulted in the identification of 12 core Al-stress responsive proteins conserved between Al stress conditions with or without S. luteus inoculation. These proteins are involved in chaperonin CPN60-2, protein refolding and ATP-binding, Cu-Zn-superoxide dismutase precursor, oxidation-reduction process, and metal ion binding, phosphoglycerate kinase 1, glycolytic process, and metabolic process. Furthermore, 198 Al responsive proteins were identified specifically under S. luteus-inoculation and are involved in gene regulation, metabolic process, oxidation-reduction process, hydrolase activity, and peptide activity. Chlorophyll a-b binding protein, endoglucanase, putative spermidine synthase, NADH dehydrogenase, and glutathione-S-transferase were found with a significant positive expression under a combined Al and S. luteus treatment, further supported by the up-regulation of their corresponding genes. This study provides a theoretical foundation for exploiting the regulatory role of ectomycorrhizal inoculation and associated genetic changes in resistance against Al stress in masson pine. [ABSTRACT FROM AUTHOR]

Masson pine (Pinus massoniana) is a major fast-growing timber species planted in southern China, a region of seasonal drought. Using a drought-tolerance genotype of Masson pine, we conducted large-scale transcriptome sequencing using Illumina technology. This work aimed to evaluate the transcriptomic responses of Masson pine to different levels of drought stress. First, 3397, 1695 and 1550 unigenes with differential expression were identified by comparing plants subjected to light, moderate or severe drought with control plants. Second, several gene ontology (GO) categories (oxidation-reduction and metabolism) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways (plant hormone signal transduction and metabolic pathways) were enriched, indicating that the expression levels of some genes in these enriched GO terms and pathways were altered under drought stress. Third, several transcription factors (TFs) associated with circadian rhythms (HY5 and LHY), signal transduction (ERF), and defense responses (WRKY) were identified, and these TFs may play key roles in adapting to drought stress. Drought also caused significant changes in the expression of certain functional genes linked to osmotic adjustment (P5CS), abscisic acid (ABA) responses (NCED, PYL, PP2C and SnRK), and reactive oxygen species (ROS) scavenging (GPX, GST and GSR). These transcriptomic results provide insight into the molecular mechanisms of drought stress adaptation in Masson pine. [ABSTRACT FROM AUTHOR]