Evergreen Broad-leaved Trees Research Articles

Biomass burning CO emissions: exploring insights through TROPOMI-derived emissions and emission coefficients

Abstract. Emissions from biomass burning are a significant source of air pollution, which can adversely impact air quality and ecosystems thousands of kilometres downwind. These emissions can be estimated by a bottom-up approach that relies on fuel consumed and standardized emission factors. Emissions are also commonly derived with a top-down approach, using satellite-observed fire radiative power (FRP) as a proxy for fuel consumption. Biomass burning emissions can also be estimated directly from satellite trace gas observations, including carbon monoxide (CO). Here, we explore the potential of satellite-derived CO emission rates from biomass burning and provide new insights into the understanding of satellite-derived fire CO emissions globally, with respect to differences in regions and vegetation type. Specifically, we use the TROPOMI (Tropospheric Monitoring Instrument) high-spatial-resolution satellite datasets to derive burning CO emissions directly for individual fires between 2019 and 2021 globally. Using synthetic data (with known emissions), we show that the direct emission estimate methodology has a 34 % uncertainty for deriving CO emissions (and a total uncertainty of 44 % including wind and CO column uncertainty). From the TROPOMI-derived CO emissions, we derive biome-specific emission coefficients (emissions relative to FRP) by combining the direct emission estimates and the satellite-observed FRP from the Moderate Resolution Imaging Spectrometer (MODIS). These emission coefficients are used to establish annual top-down CO emission inventories from biomass burning, showing that Southern Hemisphere Africa has the highest CO biomass burning emissions (over 25 % of global total of 300–390 Mt(CO) yr−1 between 2003–2021), and almost 25 % of global CO biomass burning emissions are from broadleaved evergreen tree fires. A comprehensive comparison between direct estimates, top-down and bottom-up approaches, provides insight into the strengths and weaknesses of each method: FINN2.5 has higher CO emissions, by a factor between 2 and 5, than all other inventories assessed in this study. Trends over the past 2 decades are examined for different regions around the globe, showing that global CO biomass burning emissions have, on the whole, decreased (by 5.1 to 8.7 Mt(CO) yr−1), where some regions experience increased and others decreased emissions.

Xylem embolism induced by freeze-thaw and drought are influenced by different anatomical traits in subtropical montane evergreen angiosperm trees.

Subtropical evergreen broadleaved forests distributed in montane zones of southern China experience seasonal droughts and winter frost. Previously, studies have recognized that xylem anatomy is a determinant of its vulnerability to embolism caused by drought and freezing events. We hypothesized that there is a coordination of xylem resistance to freeze-thaw and drought-induced embolism for the subtropical montane evergreen broadleaved tree species because they are influenced by common xylem structural traits (e.g., vessel diameter). We examined the branch xylem anatomy, resistance to drought-induced embolism (P50), and the percent loss of branch hydraulic conductivity after a severe winter frost (PLCwinter) for 15 evergreen broadleaved tree species in a montane forest in South China. Our results showed that P50 of the studied species ranged from -2.81 to -5.13 MPa, which was not associated with most xylem anatomical properties except for the axial parenchyma-to-vessel connectivity. These tree species differed substantially in PLCwinter, ranging from 0% to 76.41%. PLCwinter was positively related to vessel diameter and negatively related to vessel density, vessel group index, and vessel-to-vessel connectivity, but no coordination with P50. This study suggests that hydraulic adaptation to frost is important to determine the distributional limit of subtropical montane evergreen woody angiosperms.

Tree-Climbing Behavior of a Forest-Dwelling Ungulate: The Formosan Serow.

Ungulates are terrestrial herbivores, basically adapted to running fast on the ground; tree-climbing behavior has been reported only in seven species, and five of them live in open habitats (Capra hircus, C. aegagrus, C. falconeri, C. cylindricornis, Oreotragus oreotragus). Tree-climbing behavior may also be evolved in ungulates inhabiting dense forests with abundant trees; however, this has rarely been reported in such species (Moschus leucogaster, M. moschiferus), probably due to the difficulty of observing in the wild. The numerous publicly available records in social networks hold potentially valuable information on the atypical behaviors of wild ungulates. Here, we explored the tree-climbing behavior of a forest-dwelling ungulate, the Formosan serow in Taiwan, a subtropical island, by extracting information from online social media platforms. We researched images and videos of Formosan serows through Facebook and YouTube and collected a total of 15 tree-climbing events. In these materials, Formosan serows climbed 10 tree species, including evergreen coniferous and broad-leaved trees, and a variety of parts, ranging in height from 0.6 to 4 m, and from branches of shrubs to trunks of tall trees. Tree-climbing behavior was recorded throughout Taiwan and from lowlands to subalpine zones, suggesting that tree climbing may be a common behavior in this species. Foraging while climbing trees was frequently observed (53.3%), suggesting that the purpose or benefit for climbing is to obtain additional food other than plants growing near the ground surface. In contrast to other tree-climbing ungulates, Formosan serows climbed trees not only in winter, but also in other seasons, when food is relatively abundant. This is the first scientific report of tree-climbing behavior in the Formosan serow that is typically a forest dweller.

Single-nucleotide polymorphisms and copy number variations drive adaptive evolution to freezing stress in a subtropical evergreen broad-leaved tree: Hexaploid wild Camellia oleifera

Subtropical evergreen broad-leaved trees are usually vulnerable to freezing stress, while hexaploid wild Camellia oleifera shows strong freezing tolerance. As a valuable genetic resource of woody oil crop C. oleifera, wild C. oleifera can serve as a case for studying the molecular bases of adaptive evolution to freezing stress. Here, 47 wild C. oleifera from 11 natural distribution sites in China and 4 relative species of C. oleifera were selected for genome sequencing. “Min Temperature of Coldest Month” (BIO6) had the highest comprehensive contribution to wild C. oleifera distribution. The population genetic structure of wild C. oleifera could be divided into two groups: in cold winter (BIO6 ≤ 0 °C) and warm winter (BIO6 > 0 °C) areas. Wild C. oleifera in cold winter areas might have experienced stronger selection pressures and population bottlenecks with lower Ne than those in warm winter areas. 155 single-nucleotide polymorphisms (SNPs) were significantly correlated with the key bioclimatic variables (106 SNPs significantly correlated with BIO6). Twenty key SNPs and 15 key copy number variation regions (CNVRs) were found with genotype differentiation > 50% between the two groups of wild C. oleifera. Key SNPs in cis-regulatory elements might affect the expression of key genes associated with freezing tolerance, and they were also found within a CNVR suggesting interactions between them. Some key CNVRs in the exon regions were closely related to the differentially expressed genes under freezing stress. The findings suggest that rich SNPs and CNVRs in polyploid trees may contribute to the adaptive evolution to freezing stress.

Predicting the Potential Distribution of Quercus oxyphylla in China under Climate Change Scenarios

Global climate changes are expected to profoundly shape species distribution. Quercus oxyphylla, a valuable evergreen broad-leaved tree species, is rigorously conserved and managed in China owing to its substantial scientific, economic, and ecological value. However, the impact of projected climate change on its future distribution and potential climatic drivers remains unclear. Here, a maximum entropy model (MaxEnt) was used to explore the distribution of Q. oxyphylla in China under current conditions and three future scenarios (SSP1-2.6, SSP2-4.5, and SSP5-8.5) for the 2050s and 2070s. We optimized the model using the ‘ENMeval’ package to obtain the best parameter combination (RM = 1, FC = LQHPT), and multiple evaluation metrics (AUC ≥ 0.9; TSS ≥ 0.6; Kappa ≥ 0.75) verified the high accuracy of the model and the reliability of the prediction results. We found the following: (1) The potential distribution of Q. oxyphylla spans across 28 provinces in China under current climatic conditions, predominantly in southern regions, with Sichuan exhibiting the largest suitable area for survival. The total suitable habitat covers 244.98 × 104 km2, comprising highly, moderately, and poorly suitable habitats of 51.66 × 104 km2, 65.98 × 104 km2, and 127.34 × 104 km2, respectively. (2) Under future climate conditions, the overall geographical boundaries of Q. oxyphylla are predicted to remain similar to the present one, with an increase of 10.29% in the 2050s and 11.31% in the 2070s. In the 2050s, the total suitable habitats for Q. oxyphylla under the three scenarios (SSP1-2.6, SSP2-4.5, and SSP5-8.5) might increase by 8.83%, 9.62%, and 12.42%, while in the 2070s they might increase by 10.39%, 17.21%, and 6.33%, respectively. (3) Moreover, the centroid of the suitable area is expected to migrate southwestward under the three scenarios in the future. (4) Annual precipitation, isothermality, and temperature annual range emerged as the main factors influencing the distribution of Q. oxyphylla, with contributions of 55.9%, 25.7%, and 13.5%, respectively. Our findings refined the spatial arrangement of Q. oxyphylla growth and revealed its climate resilience. This suggested that under climate change, Sichuan and Shaanxi are the optimal regions for cultivation and management, while appropriate conservation strategies should be formulated in Tibet and Hubei.

Growth-climate relationships of four tree species in the subtropical evergreen broad-leaved forests in Southwest China

Subtropical forests are an important carbon sink, yet our understanding on tree growth response to inter-annual climate variability remains limited. This study investigated the growth-climate relationships of four evergreen broad-leaved tree species (Stewartia pteropetiolata W. C. Cheng, Schima noronhae Reinw. ex Bl. Bijdr, Machilus gamblei King ex J. D. Hooker, and Machilus yunnanensis Lec) in the subtropical evergreen broad-leaved forests (EBFs) in the Ailao Mountains of Yunnan province, southwest China. We constructed tree ring-width residual chronologies for each species and correlated them with daily and monthly climate data. Ring-width chronologies of S. pteropetiolata and S. noronhae were positively correlated with precipitation, relative humidity, and self-calibrated Palmer drought index (scPDSI) during autumn of the previous year and spring-to-early summer of the current year, indicating that these two Theaceae species were sensitive to moisture availability. Tree radial growth of two Machilus species was primarily limited by excessive precipitation and dense cloud cover during the rainy season in this middle montane cloud forest. Moving correlations between climate variables and radial growth of all species were temporally unstable, with a weakening response to climate variability in recent decades. We highlight the dendroclimatic potential of evergreen broad-leaved tree species in moist subtropical forests. The findings offer a pertinent perspective, emphasizing the necessity of separately considering distinct ecological indicators for trees in order to obtain a comprehensive understanding of climate-growth responses within highly diverse subtropical forests.

Mapping the potential distribution of Asian elephants: Implications for conservation and human–elephant conflict mitigation in South and Southeast Asia

Asian elephants play a pivotal role in their ecosystem. Understanding the potential distribution area of this species is vital for effective conservation efforts and mitigation of human-elephant conflicts. In this study, we used the maximum entropy to simulate the potential distribution area of Asian elephants across South and Southeast Asia, leveraging Maximum Entropy (MaxEnt) and presence data sourced from the Global Biodiversity Information Facility (GBIF). The analysis revealed that the potential distribution area of Asian elephants spans 530,418 km2 (10.59% of the study area), with significant potential distribution areas observed in Indonesia (136,890 km2) and Malaysia (119,497 km2). Vegetation type emerged as the dominant environmental factor influencing model outcomes, encompassing aspects such as broadleaved evergreen tree coverage, broadleaved deciduous closed tree coverage and EVI. The potential distribution area of Asian elephants overlaps with regions inhabited by 55.25 million people, with 6.07 million people residing in highly suitable habitats. India and Malaysia have high potential for human-elephant conflict (HEC) due to the high number of people living in potential and highly suitable habitats for elephants. Bangladesh and Nepal, on the other hand, have fewer people living in these habitats suitable for elephants, but they face relatively high human population density in these areas.

Selection and Validation of Reference Genes for Gene Expression Studies in Euonymus japonicus Based on RNA Sequencing.

Euonymus japonicus is one of the most low-temperature-tolerant evergreen broad-leaved tree species in the world and is widely used in urban greening. However, there are very few molecular biology studies on its low-temperature tolerance mechanism. So far, no researcher has selected and reported on its reference genes. In this study, 21 candidate reference genes (12 traditional housekeeping genes and 9 other genes) were initially selected based on gene expression and coefficient of variation (CV) through RNA-Seq (unpublished data), and qRT-PCR was used to detect the expression levels of candidate reference genes in three different groups of samples (leaves under different temperature stresses, leaves of plants at different growth stages, and different organs). After further evaluating the expression stability of these genes using geNorm, NormFinder, Bestkeeper, and RefFind, the results show that the traditional housekeeping gene eIF5A and the new reference gene RTNLB1 have good stability in the three different groups of samples, so they are reference genes with universality. In addition, we used eIF5A and RTNLB1 as reference genes to calibrate the expression pattern of the target gene EjMAH1, which confirmed this view. This article is the first to select and report on the reference gene of E. japonicus, laying the foundation for its low-temperature tolerance mechanism and other molecular biology research.

Core microbiota drive multi-functionality of the soil microbiome in the Cinnamomum camphora coppice planting

BackgroundCinnamomum camphora (L.) Presl (C. camphora) is an evergreen broad-leaved tree cultivated in subtropical China. The use of C. camphora as clonal cuttings for coppice management has become popular recently. However, little is known about the relationship between soil core microbiota and ecosystem multi-functionality under tree planting. Particularly, the effects of soil core microbiota on maintaining ecosystem multi-functionality under C. camphora coppice planting remained unclear.Materials and methodsIn this study, we collected soil samples from three points (i.e., the abandoned land, the root zone, and the transition zone) in the C. camphora coppice planting to investigate whether core microbiota influences ecosystem multi-functions.ResultsThe result showed a significant difference in soil core microbiota community between the abandoned land (AL), root zone (RZ), and transition zone (TZ), and soil ecosystem multi-functionality of core microbiota in RZ had increased significantly (by 230.8%) compared to the AL. Soil core microbiota played a more significant influence on ecosystem multi-functionality than the non-core microbiota. Moreover, the co-occurrence network demonstrated that the soil ecosystem network consisted of five major ecological clusters. Soil core microbiota within cluster 1 were significantly higher than in cluster 4, and there is also a higher Copiotrophs/Oligotrophs ratio in cluster 1. Our results corroborated that soil core microbiota is crucial for maintaining ecosystem multi-functionality. Especially, the core taxa within the clusters of networks under tree planting, with the same ecological preferences, had a significant contribution to ecosystem multi-functionality.ConclusionOverall, our results provide further insight into the linkage between core taxa and ecosystem multi-functionality. This enables us to predict how ecosystem functions respond to the environmental changes in areas under the C. camphora coppice planting. Thus, conserving the soil microbiota, especially the core taxa, is essential to maintaining the multiple ecosystem functions under the C. camphora coppice planting.

Phylogenetic diversity patterns of woody species across different tree sizes and spatial scales in a tropical forest community on Con Dao Islands

Understanding the maintenance of diversity and the assembly of communities is a primary concern in community ecology. This study explored the phylogenetic structure of an evergreen broadleaved tree community in Con Dao National Park, Vietnam. The survey, conducted in December 2023, encompassed all tree individuals with a diameter at breast height (DBH) ≥ 2.5 cm within a 4-ha study plot. These individuals were identified by species name, and their DBH was measured. Subsequently, a community phylogenetic tree was constructed using the Phylomatic online platform. The 4-ha study plot was subdivided into there different subplots based on three spatial scales (25 m × 25 m, 50 m × 50 m, and 100 m × 100 m) and all trees were classified into three different DBH classes (2.5 cm ≤ DBH < 10 cm representing small trees; 10 cm ≤ DBH < 20 cm as medium trees; and DBH ≥ 20 cm as large trees). The net relatedness index (NRI) and net nearest taxon index (NTI) were utilized to evaluate the phylogenetic structure and infer ecological processes. Our findings revealed a consistent decrease in both NRI and NTI with increasing spatial scales and tree sizes. These results suggest an overdispersed phylogenetic structure within the community across different spatial scales and tree sizes. Additionally, negative density dependence was found to have a pronounced effect on the phylogenetic structure, with a more significant impact on tree individuals from small and medium DBH classes than large ones. This study underscores the significance of phylogenetic density dependence as a primary mechanism governing species diversity and shaping the community structure of evergreen broadleaved forests in Vietnam.

Development of the Site Index Curves for <italic>Quercus acuta</italic> Stands

Background and objective: The purpose of this study is to develop a model for estimating the site index (SI), which is the criterion for determining the site productivity of the <i>Quercus acuta</i> stand as known as Evergreen broad-leaved tree (EBLT) species.Methods: The materials used in the analysis were data from 63 sample plots, and the sample plots data were collected from the National Forest Inventory (NFI) and other sites. Since the site index is determined by the height and stand age of the dominant trees, the Schumacher model, the Chapman-Richards model, and the Weibull model were applied to derive the optimal equation for estimating the dominant tree height.Results: The three models derived the parameters and went through a statistical test. Among the three models, the Chapman-Richards model was found to be optimal considering the fitness index, bias, and residuals. By transforming this model, site index estimation equations with the reference age of 30 years were created, and site index curves were derived. According to the analysis of the collected data, the site index of the <i>Quercus acuta</i> stand in Korea was found to be in the range of 10 to 16.Conclusion: As the growth distribution of <i>Quercus acuta</i> is expected to move north due to climate change, the derivation of the site index representing site productivity will be helpful in making policy and management decisions for this species.

Establishment of a Transient Transformation Protocol in Cinnamomum camphora

Cinnamomum camphora is an excellent evergreen broad-leaved tree species with strong stress tolerance, but its molecular character revelation as well as ecological and economic value improvement were limited due to the lack of a genetic transformation system. To establish a simple and efficient transient transformation system for uncovering the molecular mechanism of plant tolerating stresses and promoting the selective breeding of good varieties, the infection method, co-cultivation time, infection solution concentration, and growth density of Agrobacterium tumefaciens containing green fluorescent protein (GFP)-based calmodulin protein 3 gene (GCaMP3) were identified by monitoring the fluorescence emitted from GCaMP3 bound to Ca2+. Meanwhile, the transient transformation effects were evaluated via cytoplasmic Ca2+ concentration variations at high temperatures of 35 °C and 40 °C. When C. camphora leaves were infected with A. tumefaciens containing GCaMP3 via injection and soaking, no significant difference was detected in the fluorescence intensity over 48 h, indicating that the two infection methods had the same transient transformation efficiency. By prolonging the co-cultivation time, the fluorescence intensity gradually increased, reached its strongest at the 48th h, and then gradually declined. For the infection solution concentration, an OD600 of 0.7 led to the strongest fluorescence intensity, with an increase of 42.2%, 13.7%, 4.2%, and 14.2%, respectively, compared to that at OD600 of 0.5, 0.6, 0.8, and 0.9. When A. tumefaciens growth density OD600 was 0.5–0.7, the strongest fluorescence intensity was detected after transient transformation. Combining these optimum conditions, GCaMP3 was transferred into C. camphora, which indicated the variations in cytoplasmic Ca2+ concentration at high temperatures, with the fluorescence intensity at 35 °C and 40 °C increasing by 12.6% and 30.6%, respectively, in contrast to that at 28 °C. Therefore, it should be an efficient transient transformation system for C. camphora, with A. tumefaciens growth density OD600 of 0.5–0.7, infection solution concentration OD600 of 0.7, and co-cultivation time of 48 h by using both injection and soak infection methods, which is beneficial for uncovering the Ca2+ signal transduction in the plant tolerating stresses and promoting its molecular biology development and selective breeding of good varieties.

Urban Treetop Detection and Tree-Height Estimation from Unmanned-Aerial-Vehicle Images

Individual tree detection for urban forests in subtropical environments remains a great challenge due to the various types of forest structures, high canopy closures, and the mixture of evergreen and deciduous broadleaved trees. Existing treetop detection methods based on the canopy-height model (CHM) from UAV images cannot resolve commission errors in heterogeneous urban forests with multiple trunks or strong lateral branches. In this study, we improved the traditional local-maximum (LM) algorithm using a dual Gaussian filter, variable window size, and local normalized correlation coefficient (NCC). Specifically, we adapted a crown model of maximum/minimum tree-crown radii and an angle strategy to detect treetops. We then removed and merged the pending tree vertices. Our results showed that our improved LM algorithm had an average user accuracy (UA) of 87.3% (SD± 4.6), an average producer accuracy (PA) of 82.8% (SD± 4.1), and an overall accuracy of 93.3% (SD± 3.9) for sample plots with canopy closures less than 0.5. As for the sample plots with canopy closures from 0.5 to 1, the accuracies were 78.6% (SD± 31.5), 73.8% (SD± 10.3), and 68.1% (SD± 12.7), respectively. The tree-height estimation accuracy reached more than 0.96, with an average RMSE of 0.61 m. Our results show that the UAV-image-derived CHM can be used to accurately detect individual trees in mixed forests in subtropical cities like Shanghai, China, to provide vital tree-structure parameters for precise and sustainable forest management.

Genetic differentiation and evolution of broad-leaved evergreen shrub and tree varieties of Daphniphyllum macropodum (Daphniphyllaceae).

Tree form evolution is an important ecological specialization for woody species, but its evolutionary process with adaptation is poorly understood, especially on the microevolutionary scale. Daphniphyllum macropodum comprises two varieties: a tree variety growing in a warm temperate climate with light snowfall and a shrub variety growing in a cool temperate climate with heavy snowfall in Japan. Chloroplast DNA variations and genome-wide single-nucleotide polymorphisms across D. macropodum populations and D. teijsmannii as an outgroup were used to reveal the evolutionary process of the shrub variety. Population genetic analysis indicated that the two varieties diverged but were weakly differentiated. Approximate Bayesian computation analysis supported a scenario that assumed migration between the tree variety and the southern populations of the shrub variety. We found migration between the two varieties where the distributions of the two varieties are in contact, and it is concordant with higher tree height in the southern populations of the shrub variety than the northern populations. The genetic divergence between the two varieties was associated with snowfall. The heavy snowfall climate is considered to have developed since the middle Quaternary in this region. The estimated divergence time between the two varieties suggests that the evolution of the two varieties may be concordant with such paleoclimatic change.

Maxent Modeling for Predicting the Potential Geographical Distribution of Castanopsis carlesii under Various Climate Change Scenarios in China

Castanopsis carlesii (Hemsl.) Hayata. 1917 is an established subtropical evergreen broad-leaved tree species with rapid growth rates and a strong plasticity to environmental changes. It is widely distributed in East Asia; however, it is unclear how climate change influences the distribution of this tree species. Based on 210 valid occurrence records and 10 environmental variables, we used maximum entropy model (Maxent) to predict its potential geographical distribution under present and three future climate scenarios (SSP126, SSP245 and SSP585) in both the 2050s and 2070s, and determined the influence of climate on the distribution of C. carlesii. The area under the curve (AUC) value of the simulated training and the test were 0.949 and 0.920, respectively, indicating an excellent forecast. The main climatic factors affecting the distribution of C. carlesii are mainly precipitation, especially that of the driest month (Bio14, 75.5%), and annual precipitation (Bio12, 14.3%); its total contribution rate is 89.8%. However, the impact of average mean temperature is lesser in comparison (Bio1, 5.7%). According to the present-day predictions, C. carlesii has a suitable habitat of 208.66 × 104 km2 across most of the tropical and subtropical regions south of the Yangtze River. The medium and high suitability areas are mainly in Taiwan, Fujian, Jiangxi, Guangdong, Hainan and Guangxi Provinces. With the climate projected to warm in the future, the distribution area of C. carlesii exhibited a tendency of northward expansion along the Qinling–Huaihe line, mainly manifested as the increase in low and medium suitable areas. The area of high-suitable areas decreased significantly under the three climate scenarios both for the 2050s and 2070s, and only a few areas showed contraction of suitable areas. Therefore, expansion areas can be used for cultivation or introduction trials, while contraction areas require enhanced preservation and collection of genetic resources. Our findings provide a theoretical basis for formulating the adaptation and protection strategies to cope with future climate change as well as theoretical guidance for the introduction, cultivation and sustainable development of C. carlesii.

Seasonal changes in gas exchange, water and macro-nutrient content differ between Citrus cultivars

Citrus is an evergreen broad-leaved fruit tree including numerous species and cultivars with an extensive global planting area. However, the relationship between physiological parameters and seasonal fruit ripening of different citrus cultivars has not been reported. With the aim to obtain information on this relationship, we selected two late-maturing (Newhall Navel and Ponkan) and two middle-maturing citrus cultivars (Fertile orange and Tarocco) and compared the seasonal patterns of foliar CO2 and H2O gas exchange, leaf water content and macro-nutrient concentrations in leaves and roots. Our results showed that seasonal changes of foliar CO2 and H2O gas exchange and water content greatly differed between cultivars at defined developmental stages, but especially during fruit ripening. For example, in December, the net rates of photosynthetic CO2 fixation (Pn) of the late-maturing cultivars were significantly higher than in middle-maturing cultivars. Seasonal fluctuations of total nitrogen (N), total phosphorus (P), and inorganic phosphorus (Pi), as well as C/N, C/P, and N/P ratios in leaves and roots differed between middle and late-maturing cultivars. These differences were most obvious at fruit ripening compared to other developmental stages and were significantly higher in leaves than in roots. We conclude that seasonal differences in foliar Pn and macro-nutrient contents between leaves of citrus cultivars are largely determined by seasonal fruit ripening. In addition, seasonal changes in root macro-nutrient contents were also affected by seasonal fruit ripening of citrus cultivars. However, this effect was much lower compared to the leaves, but still indicates particular shoot-root interactions during fruit ripening.

First Report of Colletotrichum theobromicola Causing Anthracnose on Euonymus japonicus in China.

Euonymus japonicus, a broad-leaved evergreen tree, is widely planted in the parks and landscapes in the world (Huang et al. 2016). In 2019, anthracnose lesions were found on leaves of E. japonicus with 4~15% incidence from the Wufulinglong park (116°28' E, 39°94' N) in the Haidian District of Beijing, China. Irregular chlorotic spots appeared on the surface of the leaves, then larger necrotic spots with numerous acervuli gradually formed, and finally the infected leaves dried and dropped, detracting from the aesthetic of the landscape. To identify the pathogen, six representative leaves with typical symptoms in Wufulinglong park from three plants were collected and diseased tissue was cut into 2 mm × 2 mm pieces, disinfested in 75% ethanol for 20 s, rinsed twice in sterile water, plated onto potato dextrose agar (PDA), which were then incubated at 25℃ under 12 h photoperiod. A total of 24 morphologically identical isolates were obtained from the samples. Two of these isolates were randomly selected for further analysis to confirm their identity. The cultures (HDwfll1907HY and HDwfll1908HY) were deposited in the culture collection of the Beijing Academy of Agriculture and Forestry Sciences, China. Furthermore, genomic DNA was extracted and the sequences of internal transcribed spacer (ITS) regions, calmodulin (CAL), beta tubulin (TUB2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin (ACT) and chitin synthase (CHS) were amplified using the primers ITS1/ITS4 (Gardes et al. 1993, White et al. 1990), CL1C/CL2C (Weir et al. 2012), T1/Bt2b (Glass et al. 1995, O'Donnell et al. 1997), GDF1/GDR1 (Guerber et al. 2003), ACT-512F/783R and CHS-79F/345R (Carbone et al. 1999), respectively. Newly obtained sequences were deposited into GenBank with accession numbers MZ229612~13 and MZ305422~31. Phylogenetic tree based on the assessed gene loci revealed that both strains clustered closely with C. theobromicola. The strain HDwfll1907HY was randomly selected for morphological description and pathogenicity assay. The colony was initially light gray with dense aerial mycelia on PDA, which later turn to gray with grayish green on the reverse side of plates. Conidiogenous cells were cylindrical, arising from swollen hyphae. Conidia were single-celled, hyaline, subcylindrical to clavate, often with broadly rounded ends, 13.78-21.99×3.47-5.44 µm (n=50). Both phylogenetic analysis and morphology support the identification of two strains as C. theobromicola. Pathogenicity tests were performed on three healthy one-year-old E. japonicus plants using the randomly selected isolate HDwfll1907HY. The leaves were sprayed with 20 mL of conidial suspension (107 conidia/mL), sterilized water inoculation under the same condition was used as control. All the treated plants were incubated in the incubator at 25°C and 90% relative humidity under 12 h photoperiod. After 18 d, all the leaves treated with the conidial suspension showed typical symptoms of anthracnose, similar to those in the field, whereas the control leaves remained symptomless. The disease assay was replicated three times for consistency. The same fungus was reisolated from the infected leaves and identified as C. theobromicola. This is the first report of C. theobromicola causing anthracnose on E. japonicus, which provides the foundation for the management of anthracnose on E. japonicus.

First report of Colletotrichum siamense causing leaf spot on Machilus pauhoi in China.

Machilus pauhoi Kaneh. is an excellent evergreen broad-leaved tree species widely grown in China for its ornamental and economic value (He et al. 2022). In September 2021, a leaf spot was observed on M. pauhoi plants on Guantian forest farm (27°06'15.6″N, 114°34'20.72″E) in ji' an city, Jiangxi province, China. The disease incidence was estimated to be above 20%. The symptoms began as brown irregular spots, then the spots gradually expand over time, with a gray-to-brown center and dark brown-to-black edges. Small infected tissues (3 to 5 mm2) were surface-sterilized in 70% ethanol for 30 s and 2% NaClO for 60 s, and rinsed three times with sterile water (Ju et al. 2021). Tissues were placed on potato dextrose agar (PDA) and incubated at 25°C. Pure cultures were obtained by transferring hyphal tips to new PDA plates. Twenty-two isolates of Colletotrichum ssp. were obtained (isolation frequency about 78%). Three representative single-spore isolates (PN-1, PN-4, and PN-9) were used for morphological studies and phylogenetic analyses. Colonies on the PDA of the three isolates were white to gray with cottony mycelia and grayish-white on the undersides of the culture. Conidia were single-celled, straight, hyaline, cylindrical, clavate, and measured 11.4-16.8 ×4.1-5.5 µm (13.2 ± 1.0 × 4.4 ± 0.3 µm, n = 100). Appressoria were brown to dark brown, ovoid to clavate, slightly irregular to irregular, and ranged from 5.2-8.8 × 4.1-6.2 µm (6.7 ± 0.2 × 5.1 ± 0.3 µm, n=100). Morphological features were similar to Colletotrichum gloeosporioides species complex (Weir et al. 2012). The internal transcribed spacer (ITS) regions, actin (ACT), calmodulin (CAL), beta-tubulin 2 (TUB2), chitin synthase (CHS-1), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were amplified from genomic DNA for the three isolates using primers ITS1/ITS4, ACT-512F/ACT-783R, CL1/CL2, T1/Bt2b, CHS-79F/CHS-354R and GDF/GDR (Weir et al. 2012), respectively. All sequences were deposited into GenBank (ITS, ON176154 - ON176156; ACT, ON185554 - ON185556; GAPDH, ON185563 - ON185565; TUB2, ON185566 - ON185568; CHS-1, ON185560 - ON185562; CAL, ON185557 - ON185559). A maximum likelihood and Bayesian posterior probability analyses using IQtree v. 1.6.8 and Mr. Bayes v. 3.2.6 with the concatenated sequences placed PN-1, PN-4, and PN-9 in the clade of C. siamense. Based on the multi-locus phylogeny and morphology, three isolates were identified as C. siamense. The pathogenicity of three isolates was tested on nine M. pauhoi plants, which were grown in the field. Healthy leaves were wounded with a sterile needle and inoculated with 10 µL of spore suspension (106 conidia/mL). The spore suspension of each isolate was inoculated onto six leaves. Another three plants inoculated with ddH2O served as the control (Wan et al. 2022). All the inoculated leaves were covered with plastic bags to keep them moist for 2 days (relative humidity > 80%). All the inoculated leaves showed similar symptoms to those observed in the field, whereas control leaves were asymptomatic for 7 days. C. siamense was reisolated from the lesions, whereas no fungus was isolated from control leaves. Up to now, Pestalotiopsis chamaeropis, Corynespora cassiicola and Arthrinium arundinis could infect M. pauhoi plants (Zhang et al. 2021), and cause leaf spots in China. To our knowledge, this is the first report of C. siamense causing leaf spots on M. pauhoi. This work provided crucial information for epidemiologic studies and appropriate control strategies for this newly emerging disease.

Transcriptome Analysis Reveals the Heat Stress Response Genes by Fire Stimulation in Michelia macclurei Dandy

Heat stress due to external heat sources such as fire is an ecological problem for plants. When forest plants suffer from fire, high temperatures cause an array of morphological, physiological, and biochemical changes, which affect growth and development. Michelia macclurei Dandy is an evergreen broad-leaved tree species with the characteristics of fast growth, strong adaptability, and good fire-resistance. Some studies have improved the understanding of how fire behavior affects physiology, function and mortality, but the extreme heat response genes and mechanisms need improved understanding. In this study, we conducted a fire experiment (slight and severe) and RNA-Seq in M. macclure. The de novo assembly obtained 104,052 unigenes, and 48.46% were annotated in at least one public database. Specifically, 4458 and 4810 differentially expressed genes (DEGs) were identified in slight and severe fire treatment groups, respectively. In two treatment groups, 612 unigenes were differentially expressed, which were enriched in ‘oxidoreductase activity’ in the molecular function (MF) category of Gene Ontology (GO) enrichment analysis, suggesting the core role of oxidoreductase activity in response to extremely high temperatures in M. macclurei. In KEGG enrichment analysis of DEGs, the ‘plant hormone signal transduction’ is overrepresented, suggesting that this process plays an important role during heat response in M. macclurei. In the pathways of cytokinine and salicylic acid, some vital DEGs were enriched, which were related to cell division, shoot initiation, and disease resistance, and the potential interactions during heat stress were discussed. Moreover, the DEGs linked to heat stress response were identified, including heat shock factors, stress enhanced protein, signal transduction, photosystem, and major transcription factors. The qRT-PCR examination of various tissues, expression dynamics, and treatments revealed that the genes coding for the heat shock protein HSF30, stress enhanced protein, and photosystem I reaction center subunit II exhibited particularities in leaf tissue. Genes coding for heat shock proteins displayed a distinct expression pattern between fire treatment and conventional heat stress, which could signify the distinctive function of HSPs and the mechanism of heat responses. Altogether, these may interact to respond to fire stress through alterations in cellular processes, signaling transduction, and the synthesis and degradation of response proteins in M. macclurei. The results of this study provide a crucial transcriptional profile influenced by heat stress in M. macclurei, and could be of great use to explore the fire prevention mechanisms of fire-resistant tree species.

Effects of Cinnamomum camphora coppice planting on soil fertility, microbial community structure and enzyme activity in subtropical China.

Cinnamomum camphora (C. camphora) is a broad-leaved evergreen tree cultivated in subtropical China. Currently, the use of C. camphora clonal cuttings for coppice management has become popular. However, the effects of C. camphora coppice planting on soil abiotic and biotic variances remained unclear. In this study, we collected soil from three points in the seven-year C. camphora coppice planting land: under the tree canopy (P15), between trees (P50), and abandoned land (Control) to investigate the effects of C. camphora coppice planting on soil fertility, microbial community structure and enzyme activity. The results revealed that C. camphora coppice planting significantly increased soil fertility in the point under the tree canopy (P15) and point between trees (P50), and P15 had more significant effects than P50. Meanwhile, in P15 and P50, soil bacterial, fungal alpha-diversity were improved and microbial community structures were also changed. And the changes of soil organic carbon and total nitrogen promote the transformation of soil bacterial, fungal community structures, respectively. In addition, C. camphora coppice planting significantly (p < 0.05) increased soil urease (UE), polyphenol oxidase, and peroxidase activities, while significantly decreased soil ACP activity. This study demonstrated that the C. camphora coppice planting could improve soil fertility in subtropical China, which promoted the transformation of soil microbial community from oligotrophs (K-strategist) to copiotrophs (r-strategist). Thus, this work can provide a theoretical basis for soil nutrient variation and productive management of C. camphora coppice plantation in subtropical China.