Physiological and Biochemical Dynamics of Pinus massoniana Lamb. Seedlings under Extreme Drought Stress and during Recovery

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.

Heteroblastic Foliage Affects the Accumulation of Non-Structural Carbohydrates and Biomass in Pinus massoniana (Lamb.) Seedlings

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.

Diversity and Distribution of Xylophagous Beetles from Pinus thunbergii Parl. and Pinus massoniana Lamb. Infected by Pine Wood Nematode

The vectors of pinewood nematode of Bursaphelenchus xylophilus (Steiner & Bührer, 1934) are mainly known as xylophagous beetles. Understanding the composition and distribution of these xylophagous beetles in host pine trees infected by PWN is critical to control the spread of PWN. In this study, we investigated the community structures of the xylophagous beetles in two main host trees in Fujian and Shandong, Pinus massoniana Lamb. and Pinus thunbergia Parl., in different stages of infection. All beetles were collected by dissecting the whole pine trees and then identified by their morphological characteristics and COI genes. The results showed that the diversity of xylophagous beetles was different not only between the two host pine trees but also among the different infection stages. The diversity of P. massoniana xylophagous beetles was significantly higher than that of P. thunbergii, and there were also significant differences in the different stages of PWN infection. In total, Scolytinae was the most common (53.70%), followed by Curculionidae (18.26%), Cerambycidae (16.31%), and Cleridae (6.04%). Monochamus alternatus, the most important vector of PWN, occupied a large niche and showed different aggregation positions during the three infection stages in both host trees. This result might be related to the resistance of bark beetles to host trees and competition with other xylophagous beetles. The community diversity of xylophagous beetles was jointly affected by both the infection stages of PWN and the spatial niche of xylophagous beetles. Knowledge of the diversity and competitive relationships among xylophagous beetles might help regulate the population dynamics of these beetles.

Transcriptome Analysis of Needle and Root of Pinus Massoniana in Response to Continuous Drought Stress

Pinus massoniana Lamb. is an important coniferous tree species in ecological environment construction and sustainable forestry development. The function of gene gradual change and coexpression modules of needle and root parts of P. massoniana under continuous drought stress is unclear. The physiological and transcriptional expression profiles of P. massoniana seedlings from 1a half-sibling progeny during drought stress were measured and analyzed. As a result, under continuous drought conditions, needle peroxidase (POD) activity and proline content continued to increase. The malondialdehyde (MDA) content in roots continuously increased, and the root activity continuously decreased. The needles of P. massoniana seedlings may respond to drought mainly through regulating abscisic acid (ABA) and jasmonic acid (JA) hormone-related pathways. Roots may provide plant growth through fatty acid β-oxidative decomposition, and peroxisomes may contribute to the production of ROS, resulting in the upregulation of the antioxidant defense system. P. massoniana roots and needles may implement the same antioxidant mechanism through the glutathione metabolic pathway. This study provides basic data for identifying the drought response mechanisms of the needles and roots of P. massoniana.

Photosynthetic Product Allocations to the Organs of Pinus massoniana Are Not Affected by Differences in Synthesis or Temporal Variations in Translocation Rates

Photosynthesis and the allocation of photosynthetic products are the two main factors that determine plant growth. To understand the growth and productivity of Pinus massoniana Lamb., the diurnal changes in photosynthetic rate were continuously monitored. Furthermore, the translocation and allocation of the photosynthetic products synthesized in the morning and afternoon were explored using 13C pulse labeling. The results showed that: (1) on sunny days, the diurnal variation of the net photosynthetic rate showed a “double peak” curve, with an obvious “a depression” when temperatures were highest and humidity lowest. On cloudy days, it showed an irregular “jagged” curve, which was curve consistent with the variations in photosynthetically active radiation (PAR). Meanwhile, the photosynthetic rate changed with the transient changes in environmental factors such as PAR, temperature, and humidity. (2) The mean value of the net photosynthetic rate in the morning was higher than in the afternoon, and the response of the net photosynthetic rate to environmental change (PAR, temperature, humidity, and CO2 concentration) in the morning was greater than that in the afternoon. (3) The translocation of photosynthetic products synthesized in the afternoon was faster than that in the morning. Shortly after synthesis of photosynthetic products, the translocation of products synthesized in the morning tended toward upper organs (including current-year leaves and 1-year leaves), while the translocation of products synthesized in the afternoon decreased in the upper organs. However, after 15 days of 13C pulse labeling, the allocation of the photosynthetic products synthesized in the morning and afternoon tended to be the same. These results indicate that the differences in the photosynthetic products synthesized and the temporal differences in the translocation rates did not affect the final allocation of the photosynthetic products in the various organs of the P. massoniana. These results improve our knowledge of the functional phases of P. massoniana during the diurnal cycle.

Identification, classification, and characterization of AP2/ERF superfamily genes in Masson pine (Pinus massoniana Lamb.)

Transcription factors (TFs) play crucial regulatory roles in controlling the expression of the target genes in plants. APETALA2/Ethylene-responsive factors (AP2/ERF) are part of a large superfamily of plant-specific TFs whose members are involved in the control of plant metabolism, development and responses to various biotic and abiotic stresses. However, the AP2/ERF superfamily has not been identified systematically in Masson pine (Pinus massoniana), which is one of the most important conifer in southern China. Therefore, we performed systematic identification of the AP2/ERF superfamily using transcriptome sequencing data from Masson pine. In the current study, we obtained 88 members of the AP2/ERF superfamily. All PmAP2/ERF members could be classified into 3 main families, AP2 (7 members), RAV (7 members), ERF (73 members) families, and a soloist protein. Subcellular localization assays suggested that two members of PmAP2/ERF were nuclear proteins. Based on pine wood nematode (PWN) inoculated transcriptome and qPCR analysis, we found that many members of PmAP2/ERF could respond to PWN inoculation and PWN related treatment conditions in vitro. In general, members of the AP2/ERF superfamily play an important role in the response of Masson pine responds to PWN. Furthermore, the roles of the AP2/ERF superfamily in other physiological activities of Masson pine remain to be further studied.