Temporal profiling of physiological, histological, and transcriptomic dissection during auxin-induced adventitious root formation in tetraploid Robinia pseudoacacia micro-cuttings.

MAIN CONCLUSION: Optimal levels of indole-3-butyric acid (IBA) applied at the stem base promote adventitious root (AR) initiation and primordia formation, thus promoting the rooting of leafy micro-cuttings of tetraploid Robinia pseudoacacia. Tetraploid Robinia pseudoacacia L. is a widely cultivated tree in most regions of China that has a hard-rooting capability, propagated by stem cuttings. This study utilizes histological, physiological, and transcriptomic approaches to explore how root primordia are induced after indole butyric acid (IBA) treatment of micro-cuttings. IBA application promoted cell divisions in some cells within the vasculature, showing subcellular features associated with adventitious root (AR) founder cells. The anatomical structure explicitly showed that AR initiated from the cambium layer and instigate the inducible development of AR primordia. Meanwhile, the hormone data showed that similar to that of indole-3-acetic acid, the contents of trans-zeatin and abscisic acid peaked at early stages of AR formation and increased gradually in primordia formation across the subsequent stages, suggesting their indispensable roles in AR induction. On the contrary, 24-epibrassinolide roughly maintained at extremely high levels during primordium initiation thoroughly, indicating its presence was involved in cell-specific reorganization during AR development. Furthermore, antioxidant activities transiently increased in the basal region of micro-cuttings and may serve as biochemical indicators for distinct rooting phases, potentially aiding in AR formation. Transcriptomic analysis during the early stages of root formation shows significant downregulation of the abscisic acid and jasmonate signaling pathways, while ethylene and cytokinin signaling seems upregulated. Network analysis of genes involved in carbon metabolism and photosynthesis indicates that the basal region of the micro-cuttings undergoes rapid reprogramming, which results in the breakdown of sugars into pyruvate. This pyruvate is then utilized to fuel the tricarboxylic acid cycle, thereby sustaining growth through aerobic respiration. Collectively, our findings provide a time-course morphophysiological dissection and also suggest the regulatory role of a conserved auxin module in AR development in these species.

Genetically-clustered antifungal phytocytokines and receptor protein family members cooperate to trigger plant immune signaling.

Phytocytokines regulate plant immunity by cooperating with cell-surface proteins. Populus trichocarpa RUST INDUCED SECRETED PEPTIDE 1 (PtRISP1) exhibits an elicitor activity in poplar, as well as a direct antimicrobial activity against rust fungi. PtRISP1 gene directly clusters with a gene encoding a leucine-rich repeat receptor protein (LRR-RP), that we termed RISP-ASSOCIATED LRR-RP (PtRALR). In this study, we used phylogenomics to characterize the RISP and RALR gene families, and molecular physiology assays to functionally characterize RISP/RALR pairs. Both RISP and RALR gene families specifically evolved in Salicaceae species (poplar and willow), and systematically cluster in the genomes. Despite a low sequence identity, Salix purpurea RISP1 (SpRISP1) shows properties and activities similar to PtRISP1. Both PtRISP1 and SpRISP1 induced a reactive oxygen species (ROS) burst and mitogen-activated protein kinases (MAPKs) phosphorylation in Nicotiana benthamiana leaves expressing the respective clustered RALR. PtRISP1 also triggers a rapid stomatal closure in poplar. Altogether, these results suggest that plants evolved phytocytokines with direct antimicrobial activities, and that the genes coding these phytocytokines co-evolved and physically cluster with genes coding LRR-RPs required to initiate immune signaling.

Woody plant encroachment drives population declines in 20% of common open ecosystem bird species.

Grassy ecosystems cover more than 40% of the world's terrestrial surface, supporting crucial ecosystem services and unique biodiversity. These ecosystems have experienced major losses from conversion to agriculture with the remaining fragments threatened by global change. Woody plant encroachment, the increase in woody cover threatening grassy ecosystems, is a major global change symptom, shifting the composition, structure, and function of plant communities with concomitant effects on all biodiversity. To identify generalisable impacts of encroachment on biodiversity, we urgently need broad-scale studies on how species respond to woody cover change. Here, we make use of bird atlas, woody cover change data (between 2007 and 2016) and species traits, to assess: (1) population trends and woody cover responses using dynamic occupancy models; (2) how outcomes relate to habitat, diet and nesting traits; and (3) predictions of future occupancy trends, for 191 abundant, southern African bird species. We found that: (1) 63% (121) of species showed a decline in occupancy, with 18% (34) of species' declines correlated with increasing woody cover (i.e. losers). Only 2% (4) of species showed increasing population trends linked with increased woody cover (i.e. winners); (2) Open habitat specialist, invertivorous, ground nesting birds were the most frequent losers, however, we found no definitive evidence that the selected traits could predict outcomes; and (3) We predict open habitat loser species will take on average 52 years to experience 50% population declines with current rates of encroachment. Our results bring attention to concerning region-wide declining bird population trends and highlight woody plant encroachment as an important driver of bird population dynamics. Importantly, these findings should encourage improved management and restoration of our remaining grassy ecosystems. Furthermore, our findings show the importance of lands beyond protected areas for biodiversity, and the urgent need to mitigate the impacts of woody plant encroachment on bird biodiversity.

Insight of endophytic fungi promoting the growth and development of woody plants.

Microorganisms play an important role in plant growth and development. In particular, endophytic fungi is one of the important kinds of microorganisms and has a mutually beneficial symbiotic relationship with host plants. Endophytic fungi have many substantial benefits to host plants, especially for woody plants, such as accelerating plant growth, enhancing stress resistance, promoting nutrient absorption, resisting pathogens and etc. However, the effects of endophytic fungi on the growth and development of woody plants have not been systematically summarized. In this review, the functions of endophytic fungi for the growth and development of woody plants have been mainly reviewed, including regulating plant growth (e.g., flowering, root elongation, etc.) by producing nutrients and plant hormones, and improving plant disease, insect resistance and heavy metal resistance by producing secondary metabolites. In addition, the diversity of endophytic fungi could improve the ability of woody plants to adapt to adverse environment. The components produced by endophytic fungi have excellent potential for the growth and development of woody plants. This review has systematically discussed the potential regulation mechanism of endophytic fungi regulating the growth and development of woody plants, it would be of great significance for the development and utilization of endophytic fungi resource from woody plants for the protection of forest resources.

Responses of streamflow to forest expansion in a typical subhumid watershed under future climate conditions.

Water availability in the subhumid region is highly vulnerable to frequent droughts. Water scarcity in this region has become a limiting factor for ecosystem health, human livelihood, and regional economic development. A notable pattern of land cover change in the subhumid region of the United States is the increasing forest area due to afforestation/reforestation and woody plant encroachment (WPE). Given the distinct hydrological processes and runoff generation between forests and grasslands, it is important to evaluate the impacts of forest expansion on water resources, especially under future climate conditions. In this study, we focused on a typical subhumid watershed in the United States - the Little River Watershed (LRW). Utilizing SWAT + simulations, we projected streamflow dynamics at the end of the 21st century in two climate scenarios (RCP45 and RCP85) and eleven forest expansion scenarios. In comparison to the period of 2000-2019, future climate change during 2080-2099 will increase streamflow in the Little River by 5.1% in the RCP45 but reduce streamflow significantly by 30.1% in the RCP85. Additionally, our simulations revealed a linear decline in streamflow with increasing forest coverage. If all grasslands in LRW were converted into forests, it would lead to an additional 41% reduction in streamflow. Of significant concern is Lake Thunderbird, the primary reservoir supplying drinking water to the Oklahoma City metropolitan area. Our simulation showed that if all grasslands were replaced by forests, Lake Thunderbird during 2080-2099 would experience an average of 8.6 years in the RCP45 and 9.4 years in the RCP85 with water inflow amount lower than that during the extreme drought event in 2011/2012. These findings hold crucial implications for the formulation of policies related to afforestation/reforestation and WPE management in subhumid regions, which is essential to ensuring the sustainability of water resources.

Living at the edge: the functional niche occupation of woody plant communities in the submediterranean ecotone.

BACKGROUND AND AIMS: Submediterranean areas are rich ecotones, where slight modifications in environmental conditions can lead to substantial changes in the composition of plant communities. They thus offer an ideal scenario to examine plant community assembly. In this study, we followed a trait-based approach including intraspecific variability to elucidate (1) the relationship between niche occupancy components and species richness, (2) the processes governing the assembly of these communities and (3) the contribution of intraspecific trait variability in shaping the functional trait space. METHODS: We measured eight morphological and chemical traits in 405 individuals across 60 plots located in different forest communities (Mediterranean, Eurosiberian and Mixed) coexisting within a submediterranean ecosystem in central Spain. We calculated three niche occupancy components related to Hutchinson's n-dimensional hypervolumes: the total functional volume of the community, the functional overlap between species within the community and the average functional volume per species, and then used null models to explore the relative importance of habitat filtering, limiting similarity and intraspecific variability as assembly patterns. KEY RESULTS: Both habitat filtering and niche differentiation drive the community assembly of Mediterranean communities, whereas limiting similarity and hierarchical competition shape Eurosiberian communities. Intraspecific responses were mostly explained by shifts in species niches across the functional space (changes in the position of the centroids of hypervolumes). CONCLUSIONS: Different assembly mechanisms govern the structure of Mediterranean, Eurosiberian and Mixed plant communities. Combining niche occupancy components with a null model approach at different spatial scales offers new insights into the mechanisms driving plant community assembly. Consideration of intraspecific variability is key for understanding the mechanisms governing species coexistence in species-rich ecotones.

Combating an Invasive Boxwood Pathogen - Calonectria pseudonaviculata - in the United States by Shifting Production to Less-Susceptible Cultivars.

Boxwood blight (BB) caused by Calonectria pseudonaviculata (Cps), an economically devastating disease affecting everyone in the supply chain from growers to gardeners, was first officially documented in the United States in 2011. This disease has taken a heavy toll on boxwood, an iconic landscape plant and the number one evergreen nursery crop. Instead of abandoning boxwood production and switching to other evergreens, growers in the United States, informed by the latest research, have elected to combat this disease. One of the strategies employed has been to shift boxwood production from highly susceptible to less-susceptible cultivars. The objective of this study was to investigate the ongoing shift by comparing boxwood sales of 17 selected nurseries from seven states across the country in 2011, 2016, and 2021. Results revealed an additional 21.24% (from 38.58 to 60.04%) in sales of less-susceptible boxwood cultivars in 2021 compared with 2016. The less-susceptible cultivars responsible for increased boxwood sales are 'Winter Gem', 'Wintergreen', 'SB 300' (Freedom), 'SB 108' (Independence), and 'Little Missy'. The potential for long-term positive impact on sustainable boxwood production and plantings in the United States through the use of less-susceptible cultivars is discussed. This shift in boxwood choices builds crop health into new plantings of this landmark plant and sustains growth in demand for boxwood. This sets a new example of sustainable protection for a crop that is under serious pressure from an invasive pathogen.

Homogenization characteristics and regional effects in the diversity pattern of woody plants in 101 cities in China.

Influenced by the interplay of global climate change and urbanization, urban plants have become increasingly homogenized in China. However, regional effects of biotic homogenization cannot be clearly explained due to the lack of continuous large-scale data. Thus, we explored the characteristics and regional effects of biotic homogenization, which not only contributes to the improvement of urban biodiversity, but also has important value for human well-being. Here, we analyzed the woody plants of 101 cities in 8 major urban agglomerations in China. The diversity patterns and influencing factors were explored using generalized additive, geographically weighted regression, and structural equation models. The main results were as follows: (1) The issue of woody plant homogenization is primarily manifested in urban greening species in China. (2) The characteristics of woody plant homogenization exhibit notable regional effects at a large scale. (3) Latitude, urban area, altitude and climatic factors all impact the woody plant homogenization. Thus, we found that the homogenization characteristics of urban greening species exhibit regional variations, influenced by both natural and anthropogenic factors. Finally, we suggested that urban biodiversity management should be considered specific regional environmental, both to meet the needs of residents.

MxMPK4-1 phosphorylates NADPH oxidase to trigger the MxMPK6-2-MxbHLH104 pathway mediated Fe deficiency responses in apple.

Iron (Fe) deficiency is a nutritional stress in plants that commonly occurs in alkaline and calcareous soils. Mitogen-activated protein kinases (MPKs), the terminal player of MAPK cascade, are involved in distinct physiological processes. Once plants suffer from Fe deficiency stress, the mechanism of MPK function remains unclear owing to limited study on the MPK networks including substrate proteins and downstream pathways. Here, the MAP kinase MPK4-1 was induced in roots of Fe efficient apple rootstock Malus xiaojinensis but not in Fe inefficient rootstock Malus baccata under Fe deficiency conditions. Overexpression of MxMPK4-1 in apple calli and apple roots enhanced the responses to Fe deficiency. We found that MxMPK4-1 interacted with NADPH oxidases (NOX)-respiratory burst oxidase homologs MxRBOHD1 and MxRBOHD2, which positively regulated responses to Fe deficiency. Moreover, MxMPK4-1 phosphorylated the C terminus of MxRBOHD2 at Ser797 and Ser906 and positively and negatively regulated NOX activity through these phospho-sites, respectively. When compared with apple calli that overexpressed MxRBOHD2, the coexpression of MxMPK4-1 and MxRBOHD2 prominently enhanced the Fe deficiency responses. We also demonstrated that hydrogen peroxide derived from MxMPK4-1-MxRBOHD2 regulated the MxMPK6-2-MxbHLH104 pathway, illuminating a systematic network that involves different MPK proteins in M. xiaojinensis under Fe deficiency stress.

Woody-plant encroachment: Precipitation, herbivory, and grass-competition interact to affect shrub recruitment.

Woody-plant encroachment is a global phenomenon that has been affecting the southwestern United States since the late 1800s. Drought, overgrazing, herbivory, and competition between grasses and shrub seedlings have been hypothesized as the main drivers of shrub establishment. However, there is limited knowledge about the interactions among these drivers. Using a rainfall manipulation system and various herbivore exclosures, we tested hypotheses about how precipitation (PPT), competition between grasses and shrub seedlings, and predation affect the germination and first-year survival of mesquite (Prosopis glandulosa), a shrub that has encroached in Southern Great Plains and Chihuahuan Desert grasslands. We found that mesquite germination and survival (1) increased with increasing PPT, then saturated at about the mean growing season PPT level, (2) that competition between grasses and shrub seedlings had no effect on either germination or survival, and (3) that herbivory by small mammals decreased seedling establishment and survival, while ant granivory showed no effect. In addition to its direct positive effect on survival, PPT had an indirect negative effect via increasing small mammal activity. Current models predict a decrease in PPT in the southwestern United States with increased frequency of extreme events. The non-linear nature of PPT effects on Mesquite recruitment suggests asymmetric responses, wherein drought has a relatively greater negative effect than the positive effect of wet years. Indirect effects of PPT, through its effects on small mammal abundance, highlight the importance of accounting for interactions between biotic and abiotic drivers of shrub encroachment. This study provides quantitative basis for developing tools that can inform effective shrub management strategies in grasslands and savannas.

Tracking spatial regimes as an early warning for a species of conservation concern.

In this era of global environmental change and rapid regime shifts, managing core areas that species require to survive and persist is a grand challenge for conservation. Wildlife monitoring data are often limited or local in scale. The emerging ability to map and track spatial regimes (i.e., the spatial manifestation of state transitions) using advanced geospatial vegetation data has the potential to provide earlier warnings of habitat loss because many species of conservation concern strongly avoid spatial regime boundaries. Using 23 yr of data for the lek locations of Greater Prairie-Chicken (Tympanuchus cupido; GPC) in a remnant grassland ecosystem, we demonstrate how mapping changes in the boundaries between grassland and woodland spatial regimes provide a spatially explicit early warning signal for habitat loss for an iconic and vulnerable grassland-obligate known to be highly sensitive to woody plant encroachment. We tested whether a newly proposed metric for the quantification of spatial regimes captured well-known responses of GPC to woody plant expansion into grasslands. Resource selection functions showed that the grass:woody spatial regime boundary strength explained the probability of 80% of relative lek occurrence, and GPC strongly avoided grass:woody spatial regime boundaries at broad scales. Both findings are consistent with well-known expectations derived from GPC ecology. These results provide strong evidence for vegetation-derived delineations of spatial regimes to serve as generalized signals of early warning for state transitions that have major consequences to biodiversity conservation. Mapping spatial regime boundaries over time provided interpretable early warnings of habitat loss. Woody plant regimes displaced grassland regimes starting from the edges of the study area and constricting inward. Correspondingly, the relative probability of lek occurrence constricted in space. Similarly, the temporal trajectory of spatial regime boundary strength increased over time and moved closer to the observed limit of GPC lek site usage relative to grass:woody boundary strength. These novel spatial metrics allow managers to rapidly screen for early warning signals of spatial regime shifts and adapt management practices to defend and grow habitat cores at broad scales.

Tolerance capacities of Broussonetia papyrifera to heavy metal(loid)s and its phytoremediation potential of the contaminated soil.

Broussonetia papyrifera, is a promising fast-growing woody plant for the phytoremediation of heavy metal(loid) (HM)-contaminated soil. In this study, a greenhouse experiment was conducted to explore the tolerance capacities of B. papyrifera and its phytoremediation potential in the HM-contaminated soil. The results indicated that B. papyrifera could effectively decrease malondialdehyde (MDA) content by enhancing the antioxidant enzyme activities along with the cultivation in the HM-contaminated soil. Significant (p < 0.05) negative relationships were found between MDA content and superoxide dismutase (r = -0.620) and catalase activities (r = -0.702) in B. papyrifera leaves. Fourier Transform Infrared Spectroscopy analysis indicated that the main functional groups in B. papyrifera roots were slightly influenced by HMs, and organic acids, carbohydrates, protein, and amino acids might bind with HMs in plant roots to alleviate the adverse effect of HMs on plants growth. Meanwhile, B. papyrifera had great potential used for the phytoextraction of Cd and Zn in HM-contaminated soil. The maximum total Cd and Zn accumulation amount in B. papyrifera shoots could attach to 2.26 and 66.8 mg·pot-1, respectively. These observations suggested that B. papyrifera has large biomass and high tolerance to HMs, which can be regarded as a promising plant for the eco-remediation of HM-contaminated sites.Novelty statement In this study, a fast-growing woody plant, Broussonetia papyrifera, was used for heavy metal(loid) (HM)-contaminated soil remediation. We found that B. papyrifera can effectively alleviate the adverse effect of HMs on plant growth by enhancing the antioxidant enzyme activities in leaves and binding HMs with organic acids, carbohydrates, protein, and amino acids in roots. Furthermore, the maximum total Cd and Zn accumulation amount in B. papyrifera shoots could attach to 2.26 and 66.8 mg·pot-1, which suggested that B. papyrifera might be regarded as a promising woody plant used for the phytoextraction of Cd and Zn in the contaminated soil.

Insect Floral Visitors of Ptelea trifoliata (Rutaceae) in Iowa, United States.

Ptelea trifoliata L., is a North American tree that supports insect communities through floral rewards. Our objectives were to determine the importance of insects as pollinators of P. trifoliata; describe the community of floral visiting insects of P. trifoliata in Iowa, where no such information was available; and to note insect preferences for male or female flowers. Over two years, inflorescences on 13 trees were covered with mesh bags before blooming and the amount of fruit produced was compared to uncovered inflorescences from the same trees. In one year, insects were collected from male and female trees with an insect vacuum every 3 h between 7 am and 7 pm from four sites in Iowa, USA between 30 May and 16 June 2020. In 2019 and 2020, almost no fruit set occurred from inflorescences covered with mesh bags while an average of 51.2 fruits formed on unbagged inflorescences (P < 0.0001), suggesting insects larger than the 600 µm pore diameters mesh were responsible for pollination of P. trifoliata. Insects from five orders, 49 families, and at least 109 species were collected. Most insects were Hymentoptera (48.3%) or Diptera (28.2%). Male flowers attracted 62.3% of all insects collected. Since most of the insects found visiting P. trifoliata were not bees, the floral rewards of the flowers may be a valuable resource for a wide variety of insects in the central United States.

Identification and Investigation of the Genetic Variations and Candidate Genes Responsible for Seed Weight via GWAS in Paper Mulberry.

Seeds directly determine the survival and population size of woody plants, but the genetic basis of seed weight in woody plants remain poorly explored. To identify genetic variations and candidate genes responsible for seed weight in natural woody populations, we investigated the hundred-seed weight of 198 paper mulberry individuals from different areas. Our results showed that the hundred-seed weight of paper mulberry was significantly associated with the bioclimatic variables of sampling sites, which increased from south to north along the latitudinal-temperature gradient. Using 2,414,978 high-quality SNPs from re-sequencing data, the genome-wide association analysis of the hundred-seed weight was performed under three models, which identified 148, 19 and 12 associated genes, respectively. Among them, 25 candidate genes were directly hit by the significant SNPs, including the WRKY transcription factor, fatty acid desaturase, F-box protein, etc. Most importantly, we identified three crucial genetic variations in the coding regions of candidate genes (Bp02g2123, Bp01g3291 and Bp10g1642), and significant differences in the hundred-seed weight were detected among the individuals carrying different genotypes. Further analysis revealed that Bp02g2123 encoding a fatty acid desaturase (FAD) might be a key factor affecting the seed weight and local climate adaptation of woody plants. Furthermore, the genome-wide investigation and expression analysis of FAD genes were performed, and the results suggested that BpFADs widely expressed in various tissues and responded to multiple phytohormone and stress treatments. Overall, our study identifies valuable genetic variations and candidate genes, and provides a better understanding of the genetic basis of seed weight in woody plants.

Evaluation of ammonia and nitrate distribution and reduction within stormwater green infrastructure with different woody plants under multiple influencing factors.

The impact of stormwater green infrastructures (GIs) with different woody plants on nitrogen (N) distribution is still poorly understood. Laboratory experiments were conducted for GIs without or with Sophora japonica and Malus baccata to investigate the distribution of NH3-N and NO3-N. The test data was utilized to calibrate and validate the HYDRUS-2D. The validated model was subsequently used to analyze the distribution of NH3-N and NO3-N within the different GIs under three different rainfall conditions: inflow/runoff pollutant concentration, rainfall recurrence interval (runoff amount of a rainfall event), and number of dry days (during which no rainwater infiltrates into the soil). The average NH3-N and NO3-N concentrations in the upper soil (0-30 cm) of the GIs were about 4.8 and 2.4 times those of the lower layer (30-60 cm). Compared to the control (Vc), the average NH3-N concentrations in soil with Sophora japonica (Vs) and Malus baccata (Vm) decreased by 15.8% and 35.1% while those of NO3-N decreased by 15.5% and 27.2%, respectively. Degrees of influence by the three factors on the average soil NH3-N and NO3-N concentrations were inflow concentration > number of dry days > recurrence interval. The number of dry days was the smallest influence factor for the overflow N load while the inflow concentration was the most significant influence factor for the outflow, bio-utilization, and soil nitrogen loads. Compared to the control, outflow (groundwater recharge) loads of NO3-N from the Vs and Vm increased by 14.0-16.6% and 3.7-6.8%, respectively under different conditions. The overflow (runoff) loads from Vs and Vm decreased by 16.8-36.3% and 6.6%-8.4%, respectively. A multiple regression equation was used to establish a quantitative coupling relationship between N pollutant load reduction rates and influence factors (R2 ≥ 0.83). This relationship can be used to estimate the runoff treatment effectiveness of green infrastructure on target pollutants.

Natural population re-sequencing detects the genetic basis of local adaptation to low temperature in a woody plant.

KEY MESSAGE: Total of 14 SNPs associated with overwintering-related traits and 75 selective regions were detected. Important candidate genes were identified and a possible network of cold-stress responses in woody plants was proposed. Local adaptation to low temperature is essential for woody plants to against changeable climate and safely survive the winter. To uncover the specific molecular mechanism of low temperature adaptation in woody plants, we sequenced 134 core individuals selected from 494 paper mulberry (Broussonetia papyrifera), which naturally distributed in different climate zones and latitudes. The population structure analysis, PCA analysis and neighbor-joining tree analysis indicated that the individuals were classified into three clusters, which showed forceful geographic distribution patterns because of the adaptation to local climate. Using two overwintering phenotypic data collected at high latitudes of 40°N and one bioclimatic variable, genome-phenotype and genome-environment associations, and genome-wide scans were performed. We detected 75 selective regions which possibly undergone temperature selection and identified 14 trait-associated SNPs that corresponded to 16 candidate genes (including LRR-RLK, PP2A, BCS1, etc.). Meanwhile, low temperature adaptation was also supported by other three trait-associated SNPs which exhibiting significant differences in overwintering traits between alleles within three geographic groups. To sum up, a possible network of cold signal perception and responses in woody plants were proposed, including important genes that have been confirmed in previous studies while others could be key potential candidates of woody plants. Overall, our results highlighted the specific and complex molecular mechanism of low temperature adaptation and overwintering of woody plants.

The use of PacBio SMRT technology to explore the microbial network and fermentation characteristics of woody silage prepared with exogenous carbohydrate additives.

AIMS: To effectively use woody plant resources to prepare silage for ruminants, Pacific Biosciences (PacBio) single-molecule real-time (SMRT) sequencing was applied to study the microbial network and fermentation characteristics of paper mulberry (PM) silage prepared with corn meal (CM) and rice bran (RB) as exogenous additives. METHODS AND RESULTS: PM is rich in nutrients and contains more than 26% crude protein in dry matter. After ensiling, the microbial diversity and abundance in PM, CM and RB decreased due to the anaerobic environment and acidic conditions. The CM-treated PM silage accelerated the conversion of the dominant microbial community from harmful bacteria to lactic acid bacteria and promoted lactic acid fermentation. When RB was used to treat PM silage, Enterobacter and Clostridium species became the main bacterial community during ensiling, leading to butyric acid fermentation and protein decomposition. Compared with RB, CM increased the amount of fermentation substrates, changed the microbial community structure and affected metabolic pathways (global metabolism, carbohydrate metabolism and amino acid metabolism), which improved the flavour and quality of the PM silage. CONCLUSIONS: The CM addition of improved the fermentation quality of PM silage, with PM + CM being the ideal combination. The SMRT sequencing technology could accurately obtain specific details of the microbial networks and fermentation characteristics. Our results indicate that PM can be used as a potential high-protein silage in animal production. SIGNIFICANCE AND IMPACT OF THE STUDY: In tropics, the effective use of abundant natural biomass resources such as woody plants to prepare silage for feed preservation can solve the problem of restricting livestock production due to the shortage of feed in the dry season. SMRT sequencing technology was used to accurately analyze the microbial network and fermentation characteristics of woody silage prepared with CM as an exogenous additive to improve the fermentation quality of silage.

Identification of the Optimal Season and Spectral Regions for Shrub Cover Estimation in Grasslands.

Woody plant encroachment (WPE), the expansion of native and non-native trees and shrubs into grasslands, is a less studied factor that leads to declines in grassland ecosystem health. With the increasing application of remote sensing in grassland monitoring and measuring, it is still difficult to detect WPE at its early stages when its spectral signals are not strong enough. Even at late stages, woody species have strong vegetation characteristics that are commonly categorized as healthy ecosystems. We focus on how shrub encroachment can be detected through remote sensing by looking at the biophysical and spectral properties of the WPE grassland ecosystem, investigating the appropriate season and wavelengths that identify shrub cover, testing the spectral separability of different shrub cover groups and by revealing the lowest shrub cover that can be detected by remote sensing. Biophysical results indicate spring as the best season to distinguish shrubs in our study area. The earliest shrub encroachment can be identified most likely only when the cover reaches between 10% and 25%. A correlation between wavelength spectra and shrub cover indicated four regions that are statistically significant, which differ by season. Furthermore, spectral separability of shrubs increases with their cover; however, good separation is only possible for pure shrub pixels. From the five separability metrics used, Transformed divergence and Jeffries-Matusita distance have better interpretations. The spectral regions for pure shrub pixel separation are slightly different from those derived by correlation and can be explained by the influences from land cover mixtures along our study transect.

Overcoming an "irreversible" threshold: A 15-year fire experiment.

A key pursuit in contemporary ecology is to differentiate regime shifts that are truly irreversible from those that are hysteretic. Many ecological regime shifts have been labeled as irreversible without exploring the full range of variability in stabilizing feedbacks that have the potential to drive an ecological regime shift back towards a desirable ecological regime. Removing fire from grasslands can drive a regime shift to juniper woodlands that cannot be reversed using typical fire frequency and intensity thresholds, and has thus been considered irreversible. This study uses a unique, long-term experimental fire landscape co-dominated by grassland and closed-canopy juniper woodland to determine whether extreme fire can shift a juniper woodland regime back to grassland dominance using aboveground herbaceous biomass as an indicator of regime identity. We use a space-for-time substitute to quantify herbaceous biomass following extreme fire in juniper woodland up to 15 years post-fire and compare these with (i) 15 years of adjacent grassland recovery post-fire, (ii) unburned closed-canopy juniper woodland reference sites and (iii) unburned grassland reference sites. Our results show grassland dominance rapidly emerges following fires that operate above typical fire intensity thresholds, indicating that grassland-juniper woodlands regimes are hysteretic rather than irreversible. One year following fire, total herbaceous biomass in burned juniper stands was comparable to grasslands sites, having increased from 5 ± 3 g m-2 to 142 ± 42 g m-2 (+2785 ± 812 percent). Herbaceous dominance in juniper stands continued to persist 15-years after initial treatment, reaching a maximum of 337 ± 42 g m-2 eight years post-fire. In juniper encroached grasslands, fires that operate above typical fire intensity thresholds can provide an effective method to reverse juniper woodland regime shifts. This has major implications for regions where juniper encroachment threatens rancher-based economies and grassland biodiversity and provides an example of how to operationalize resilience theory to disentangle irreversible thresholds from hysteretic system behavior.

Multiple trade-offs regulate the effects of woody plant removal on biodiversity and ecosystem functions in global rangelands.

Woody plant encroachment is a major land management issue. Woody removal often aims to restore the original grassy ecosystem, but few studies have assessed the role of woody removal on ecosystem functions and biodiversity at global scales. We collected data from 140 global studies and evaluated how different woody plant removal methods affected biodiversity (plant and animal diversity) and ecosystem functions (plant production, hydrological function, soil carbon) across global rangelands. Our results indicate that the impact of removal is strongly context dependent, varying with the specific response variable, removal method, and traits of the target species. Over all treatments, woody plant removal increased grass biomass and total groundstorey diversity. Physical and chemical removal methods increased grass biomass and total groundstorey biomass (i.e., non-woody plants, including grass biomass), but burning reduced animal diversity. The impact of different treatment methods declined with time since removal, particularly for total groundstorey biomass. Removing pyramid-shaped woody plants increased total groundstorey biomass and hydrological function but reduced total groundstorey diversity. Environmental context (e.g., aridity and soil texture) indirectly controlled the effect of removal on biomass and biodiversity by influencing plant traits such as plant shape, allelopathic, or roots types. Our study demonstrates that a one-size-fits-all approach to woody plant removal is not appropriate, and that consideration of woody plant identity, removal method, and environmental context is critical for optimizing removal outcomes. Applying this knowledge is fundamental for maintaining diverse and functional rangeland ecosystems as we move toward a drier and more variable climate.