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Abstract Miscanthus spp. show excellent application prospects due to its bioenergy potential and multiple ecological services. Annual N export with biomass harvest from Miscanthus, even without fertilizer supplement, do not reduce soil N levels. The question arises regarding how Miscanthus can maintain stable soil N levels. Metagenomic strategies were used to reveal soil N‐cycling‐related microbiome and their functional contributions to processes of soil N‐cycling based on the comparison among the bare land, cropland, 10‐year Miscanthus × giganteus, and 15‐year Miscanthus sacchariflorus fields. The results showed that, after long‐term cropland‐to‐Miscanthus conversion (LCMC), 16 of 21 bacterial phyla and all the archaeal phyla exhibited significant changes. Soil microbial denitrification and nitrification functions were significantly weakened, and N fixation (NF) was significantly enhanced. The biosynthesis of amino acids, especially alanine, aspartate, and glutamate metabolism, in soil N‐cycling‐related microbiome was dramatically promoted. The genus Anaeromyxobacter contributed largely to the NF process after LCMC. Variations in the soil available potassium, available N, organic C, and total N contents drove a functional shift of soil microbiome from cropland to Miscanthus pattern. We conclude that Miscanthus can recruit Anaeromyxobacter communities to enhance NF benefiting its biomass sustainability and soil N balance.

Polyploid plants often exhibit enhanced stress tolerance. Switchgrass is a perennial rhizomatous bunchgrass that is considered ideal for cultivation in marginal lands, including sites with saline soil. In this study, we investigated the physiological responses and transcriptome changes in the octoploid and tetraploid of switchgrass (Panicum virgatum L. ‘Alamo’) under salt stress. We found that autoploid 8× switchgrass had enhanced salt tolerance compared with the amphidiploid 4× precursor, as indicated by physiological and phenotypic traits. Octoploids had increased salt tolerance by significant changes to the osmoregulatory and antioxidant systems. The salt-treated 8× Alamo plants showed greater potassium (K+) accumulation and an increase in the K+/Na+ ratio. Root transcriptome analysis for octoploid and tetraploid plants with or without salt stress revealed that 302 upregulated and 546 downregulated differentially expressed genes were enriched in genes involved in plant hormone signal transduction pathways and were specifically associated with the auxin, cytokinin, abscisic acid, and ethylene pathways. Weighted gene co-expression network analysis (WGCNA) detected four significant salt stress-related modules. This study explored the changes in the osmoregulatory system, inorganic ions, antioxidant enzyme system, and the root transcriptome in response to salt stress in 8× and 4× Alamo switchgrass. The results enhance knowledge of the salt tolerance of artificially induced homologous polyploid plants and provide experimental and sequencing data to aid research on the short-term adaptability and breeding of salt-tolerant biofuel plants.

Soil stoichiometric characteristics and aggregate stability are affected by vegetation restoration in degraded land. Yet, it is not known that how the aggregate stability is related to soil stoichiometric characteristics under different vegetation restorations. A 5-year in situ experiment was conducted to investigate the effects of vegetation restoration on the composition, stability and stoichiometric properties of soil aggregates. In the northwest part of Beijing, Bromus inermis Leyss. and Medicago sativa L. were planted in a typical area of desertification, and natural restoration was used as control. Boosted regression trees (BRTs) were applied to partition the factors that control aggregate stability. The results showed that the mean weight diameter (MWD) of soil water-stable aggregates under natural restoration (CK) and Medicago sativa L. sowing treatments (AF) was significantly higher than that under the restoration of the Bromus inermis Leyss. sowing treatments (SB). Compared with CK, AF significantly increased the geometric mean diameter (GMD) water stable aggregates, while SB showed the opposite result. AF significantly increased the proportion of soil aggregates >2 mm compared with CK. AF could improve the stability of soil aggregates by increasing the proportion of large aggregates. For the stoichiometric characteristics of the aggregates, AF increased significantly the value of C/P in 0.053−0.125 mm particle size aggregates in all soil layers. The MBC/MBN ratio aggregates at depths of 0–10 cm and 10–20 cm was also significantly increased in the treatment of AF. The BRTs indicated that stoichiometric ratio is the main factor driving the stability of soil aggregates rather than microbial community characteristics and soil nutrients. The C/P is the main driving factor affecting the MWD, in which the overall stoichiometric influence accounts for 46%, followed by the microbial influence of 36%. For the GMD, MBC/MBN is the main driving factor, and the stoichiometric influence accounts for 94%. Our findings indicate that AF is beneficial to the stability of deep soil aggregates, and their stoichiometric characteristics of soil are the key factors affecting the stability of soil aggregates.

Pennisetum alopecuroides is an important forage grass resource, which plays a vital role in ecological environment improvement. Therefore, the acquisition of P. alopecuroides genome resources is conducive to the study of the adaptability of Pennisetum species in ecological remediation and forage breeding development. Here we assembled a P. alopecuroides cv. 'Liqiu' genome at the chromosome level with a size of approximately 845.71 Mb, contig N50 of 84.83Mb, and genome integrity of 99.13% as assessed by CEGMA. A total of 833.41-Mb sequences were mounted on nine chromosomes by Hi-C technology. In total, 60.66% of the repetitive sequences and 34,312 genes were predicted. The genomic evolution analysis showed that P. alopecuroides cv. 'Liqiu' was isolated from Setaria 7.53–13.80 million years ago and from Cenchrus 5.33–8.99 million years ago, respectively. The whole-genome event analysis showed that P. alopecuroides cv. 'Liqiu' underwent two whole-genome duplication (WGD) events in the evolution process, and the duplication events occurred at a similar time to that of Oryza sativa and Setaria viridis. The completion of the genome sequencing of P. alopecuroides cv. 'Liqiu' provides data support for mining high-quality genetic resources of P. alopecuroides and provides a theoretical basis for the origin and evolutionary characteristics of Pennisetum.

IntroductionThe stimulation of plant and microbial growth has been widely observed as a result of elevated CO2 concentrations (eCO2), however, this stimulation could be influenced by various factors and their relative importance remains unclear.MethodsA global meta-analysis was performed using 884 lines of observations collected from published papers, which analyzed the eCO2 impact on plant and microbial biomass.ResultsA significant positive impact of eCO2 was observed on various biomass measures, including aboveground biomass (20.5%), belowground biomass (42.6%), soil microbial biomass (10.4%), fungal biomass (11.0%), and bacterial biomass (9.2%). It was found that eCO2 levels above 200 ppm had a greater impact on plant biomass compared to concentrations at or below 200 ppm. On the other hand, studies showed that positive effects on microbial biomass were more prominent at lower eCO2 levels (≤200 ppm) than at higher levels (>200 ppm), which could be explained by soil nitrogen limitations. Importantly, our results indicated that aboveground biomass was controlled more by climatic and experimental conditions, while soil properties strongly impacted the stimulation of belowground and microbial biomass.DiscussionOur results provided evidence of the eCO2 fertilization effect across various ecosystem types, experimental methods, and climates, and provided a quantitative estimate of plant and soil microbial biomass sensitivity to eCO2. The results obtained in this study suggest that ecosystem models should consider climatic and edaphic factors to more accurately predict the effects of global climate change and their impact on ecosystem functions.

The relationship between metabolite changes and biological endpoints in response to cadmium (Cd) stress remains unclear. Fountain grass has good Cd enrichment and tolerance abilities and is widely used in agriculture and landscaping. We analyzed the metabolic responses by detecting the metabolites through UPLC-MS and examined the relationships between metabolite changes and the characteristics of morphology and physiology to different Cd stress in fountain grass. Our results showed that under Cd stress, 102 differential metabolites in roots and 48 differential metabolites in leaves were detected, with 20 shared metabolites. Under Cd stress, most of the carbohydrates in leaves and roots decreased, which contributed to the lowered leaf/root length and fresh weight. In comparison, most of the differential amino acids and lipids decreased in the leaves but increased in the roots. Almost all the differential amino acids in the roots were negatively correlated with root length and root fresh weight, while they were positively correlated with malondialdehyde content. However, most of the differential amino acids in the leaves were positively correlated with leaf length and leaf fresh weight but negatively correlated with malondialdehyde content. Metabolic pathway analysis showed that Cd significantly affects seven and eight metabolic pathways in the leaves and roots, respectively, with only purine metabolism co-existing in the roots and leaves. Our study is the first statement on metabolic responses to Cd stress and the relationships between differential metabolites and biological endpoints in fountain grass. The coordination between various metabolic pathways in fountain grass enables plants to adapt to Cd stress. This study provides a comprehensive framework by explaining the metabolic plasticity and Cd tolerance mechanisms of plants.

The chloroplast genomes of angiosperms usually have a stable circular quadripartite structure that exhibits high consistency in genome size and gene order. As one of the most diverse genera of angiosperms, Carex is of great value for the study of evolutionary relationships and speciation within its genus, but the study of the structure of its chloroplast genome is limited due to its highly expanded and restructured genome with a large number of repeats. In this study, we provided a more detailed account of the chloroplast genomes of Carex using a hybrid assembly of second- and third-generation sequencing and examined structural variation within this genus. The study revealed that chloroplast genomes of four Carex species are significantly longer than that of most angiosperms and are characterized by high sequence rearrangement rates, low GC content and gene density, and increased repetitive sequences. The location of chloroplast genome structural variation in the species of Carex studied is closely related to the positions of long repeat sequences; this genus provides a typical example of chloroplast structural variation and expansion caused by long repeats. Phylogenetic relationships constructed based on the chloroplast protein-coding genes support the latest taxonomic system of Carex, while revealing that structural variation in the chloroplast genome of Carex may have some phylogenetic significance. Moreover, this study demonstrated a hybrid assembly approach based on long and short reads to analyze complex chloroplast genome assembly and also provided an important reference for the analysis of structural rearrangements of chloroplast genomes in other taxa.

The halophyte Karelinia caspia has not only fodder and medical value but also can remediate saline-alkali soils. Our previous study showed that salt-secreting by salt glands is one of main adaptive strategies of K. caspia under high salinity. However, ROS scavenging, ion homeostasis, and photosynthetic characteristics responses to high salinity remain unclear in K. caspia. Here, physio-biochemical responses and gene expression associated with ROS scavenging and ions transport were tested in K. caspia subjected to 100–400 mM NaCl for 7 days. Results showed that both antioxidant enzymes (SOD, APX) activities and non-enzymatic antioxidants (chlorogenic acid, α-tocopherol, flavonoids, polyamines) contents were significantly enhanced, accompanied by up-regulating the related enzyme and non-enzymatic antioxidant synthesis gene (KcCu/Zn-SOD, KcAPX6, KcHCT, KcHPT1, Kcγ-TMT, KcF3H, KcSAMS and KcSMS) expression with increasing concentrations of NaCl. These responses are beneficial for removing excess ROS to maintain a stable level of H2O2 and O2− without lipid peroxidation in the K. caspia response to high salt. Meanwhile, up-regulating expression of KcSOS1/2/3, KcNHX1, and KcAVP was linked to Na+ compartmentalization into vacuoles or excretion through salt glands in K. caspia. Notably, salt can improve the function of PSII that facilitate net photosynthetic rates, which is helpful to growing normally in high saline. Overall, the findings suggested that ROS scavenging systems and Na+/K+ transport synergistically contributed to redox equilibrium, ion homeostasis, and the enhancement of PSII function, thereby conferring high salt tolerance.

Abstract Miscanthus spp. are increasingly cultivated in cropland worldwide due to their bioenergy potential and multiple ecological services. Effects of long‐term cropland‐to‐Miscanthus conversion without N fertilizer on soil microbiome and N cycling largely remain unknown. We aimed to explore the effects of Miscanthus conversion on soil microbiome and N cycling over a 15‐year period. We analyzed diversity, composition, and abundance of bacterial and archaeal communities using 16S rRNA amplicon sequencing, and abundances of N‐cycling‐related genes using quantitative polymerase chain reaction of 0–10 cm soils collected from bare land, cropland, 10‐year Miscanthus × giganteus, and 15‐year Miscanthus sacchriflorus land in Beijing. Conversion decreased soil sand and micro‐aggregate proportion, nitrate N (NiN), available phosphorus levels, conductivity, temperature, and pH, while increasing proportion of soil clay and macro‐aggregate (MAA), soil organic C (SOC), available N (AN), exchangeable Mg2+ (EMg2+), and available potassium (AK) contents as well as microbial C/N. Consequently, diversity, composition, and abundance of soil bacterial community exhibited larger changes than those values of archaeal community after conversion. Soil AP, EMg2+, AK, and SOC were key factors in shifting microbiome from the cropland to Miscanthus pattern. Moreover, abundances of bacterial and archaeal communities and the N fixer gene nifH increased, whereas that of the bacterial ammonia monooxygenase gene decreased. The copies of other N‐cycling‐related genes in the two Miscanthus lands seemed similar to those values of cropland. The nifH copies negatively correlated with soil NiN and positively correlated with AN, EMg2+, ECa2+, SOC, AK, and MAA. We conclude that changes in soil microbiome pattern induced by the variation of soil properties enhance microbial N fixation potential, maintaining stable N levels and robust N cycling with lower N leakage risk after conversion. These results should inspire farmers and governments to large‐scale use Miscanthus on marginal cropland in Northern China.

Abstract Background Carex L. is one of the largest genera in the Cyperaceae family and an important vascular plant in the ecosystem. However, the genetic background of Carex is complex and the classification is not clear. In order to investigate the gene function annotation of Carex, RNA-sequencing analysis was performed. Simple sequence repeats (SSRs) were generated based on the Illumina data and then were utilized to investigate the genetic characteristics of the 79 Carex germplasms. Results In this study, 36,403 unigenes with a total length of 41,724,615 bp were obtained and annotated based on GO, KOG, KEGG, NR databases. The results provide a theoretical basis for gene function exploration. Out of 8776 SSRs, 96 pairs of primers were randomly selected. One hundred eighty polymorphic bands were amplified with a polymorphism rate of 100% based on 42 pairs of primers with higher polymorphism levels. The average band number was 4.3 per primer, the average distance value was 0.548, and the polymorphic information content was ranged from 0.133 to 0.494. The number of observed alleles (Na), effective alleles (Ne), Nei’s (1973) gene diversity (H), and the Shannon information index (I) were 2.000, 1.376, 0.243, and 0.391, respectively. NJ clustering divided into three groups and the accessions from New Zealand showed a similar genetic attribute and clustered into one group. UPGMA and PCoA analysis also revealed the same result. The analysis of molecular variance (AMOVA) revealed a superior genetic diversity within accessions than between accessions based on geographic origin cluster and NJ cluster. What’s more, the fingerprints of 79 Carex species are established in this study. Different combinations of primer pairs can be used to identify multiple Carex at one time, which overcomes the difficulties of traditional identification methods. Conclusions The transcriptomic analysis shed new light on the function categories from the annotated genes and will facilitate future gene functional studies. The genetic characteristics analysis indicated that gene flow was extensive among 79 Carex species. These markers can be used to investigate the evolutionary history of Carex and related species, as well as to serve as a guide in future breeding projects.

The turfgrass species Carex rigescens has broad development and utilization prospects in landscaping construction. However, seed dormancy and a low germination rate have inhibited its application. Furthermore, the molecular mechanisms of seed germination in C. rigescens have not been thoroughly studied. Therefore, in the present study, PacBio full-length transcriptome sequencing combined with Illumina sequencing was employed to elucidate the germination mechanism of C. rigescens seeds under variable temperatures. In general, 156,750 full-length non-chimeric sequences, including those for 62,086 high-quality transcripts, were obtained using single-molecule long read sequencing. In total, 40,810 high-quality non-redundant, 1,675 alternative splicing, 28,393 putative coding sequences, and 1,052 long non-coding RNAs were generated. Based on the newly constructed full-length reference transcriptome, 23,147 differentially expressed genes were identified. We screened four hub genes participating in seed germination using weighted gene co-expression network analysis. Combining these results with the physiological observations, the important roles of sucrose and starch metabolic pathways in germination are further discussed. In conclusion, we report the first full-length transcriptome of C. rigescens, and investigated the physiological and transcriptional mechanisms of seed germination under variable temperatures. Our results provide valuable information for future transcriptional analyses and gene function studies of C. rigescens.

Pheophytinase (PPH), the phytol hydrolase, plays important roles in chlorophyll degradation. Nevertheless, little attention has been paid to the PPHs in warm-season grass species; neither its detailed function in photosynthesis has been systematically explored to date. In this study, we isolated ZjPPH from Zoysia japonica, an excellent warm-season turfgrass species. Quantitative real-time PCR analysis and promoter activity characterization revealed that the expression of ZjPPH could be induced by senescence, ABA, and dark induction. Subcellular localization observation proved that ZjPPH was localized in the chloroplasts. Overexpression of ZjPPH accelerated the chlorophyll degradation and rescued the stay-green phenotype of the Arabidopsis pph mutant. Moreover, ZjPPH promoted senescence with the accumulation of ABA and soluble sugar contents, as well as the increased transcriptional level of SAG12 and SAG14. Transmission electron microscopy investigation revealed that ZjPPH caused the decomposition of chloroplasts ultrastructure in stable transformed Arabidopsis. Furthermore, chlorophyll a fluorescence transient measurement analysis suggested that ZjPPH suppressed photosynthesis efficiency by mainly suppressing both photosystem II (PSII) and photosystem I (PSI). In conclusion, ZjPPH plays an important role in chlorophyll degradation and senescence. It could be a valuable target for genetic editing to cultivate new germplasms with stay-green performance and improved photosynthetic efficiency.

Different vegetation restoration methods may affect the soil’s physicochemical properties and microbial communities. However, it is not known how the microbial network’s complexity of the bacterial and fungal communities respond to short-term vegetation restoration. We conducted a short-term ecological restoration experiment to reveal the response of the soil’s microbial community and microbial network’s stability to initial vegetation restoration during the restoration of the degraded grassland ecosystem. The two restoration methods (sowing alfalfa (Medicago sativa, AF) and smooth brome (Bromus inermis, SB)) had no significant effect on the alpha diversity of the fungal community, but the SB significantly increased the alpha diversity of the soil surface bacterial community (p < 0.01). The results of NMDS showed that the soil’s fungal and bacterial communities were altered by a short-term vegetation restoration, and they showed that the available phosphorus (AP), available potassium (AK), and nitrate nitrogen (nitrate-N) were closely related to changes in bacterial and fungal communities. Moreover, a short-term vegetation restoration significantly increased the complexity and stability of fungi ecological networks, but the opposite was the case with the bacteria. Our findings confirm that ecological restoration by sowing may be favorable to the amelioration of soil fungi complexity and stability in the short-term. Such findings may have important implications for soil microbial processes in vegetation recovery.

Salix matsudana is an important tree species for shelterbelt and barren hill afforestation in North China. Knowledge of the seasonal pattern of the stem radial growth (SRG) of S. matsudana and its response to environmental factors is crucial for understanding the tree growth and climate relationships in the increasingly warm and dry climate of North China. To this end, a field trial was conducted in a semi-arid area of North China, and the stem radial dynamics of eight sample trees at two sites (four trees per site) with different soil moisture regimes, as well as the meteorological and edaphic conditions, were monitored from May to October in 2016 and 2019. During the growing season, the daily mean air temperature was 1.69 °C higher in 2019 than in 2016, and the total precipitation was 179.2 mm lower in 2019 than in 2016. The results showed that the seasonal pattern of S. matsudana SRG was divided into the initial stage (stage 1), the rapid growth stage (stage 2), and ending stage (stage 3) in terms of the threshold of 0.02 mm day−1 (daily growth rate). The initiation of stage 2 in 2019 (day of year (DOY) 126) was 48 days earlier than that in 2016 (DOY 174). However, the date on which the maximum growth rate was reached occurred also 34 days earlier, indicating the co-occurrence of the positive effect of the higher temperature in 2019 than in 2016 on growth initiation in stage 1 as well as the negative consequences of the water deficit on tree growth in stage 2. Higher cumulative growth of S. matsudana at the high soil moisture site was monitored relative to the low soil moisture site throughout the trial, which could be attributed to differences in soil moisture and the initial stem size of the trees in 2016 and 2019, respectively. Temperature and soil moisture were the main factors restricting daily SRG, but the growth of S. matsudana subjected to prolonged summer water stress would be less significantly limited by soil moisture in stage 3. These results showed that the seasonal pattern of SRG of S. matsudana was closely related to the dynamics of the soil and climatic factors. Therefore, seasonal variation in growth-limiting factors should be considered when forecasting tree growth in response to climate change in North China.

Abstract Background Carex L., a grass genus commonly known as sedges, is distributed worldwide and contributes constructively to turf management, forage production, and ecological conservation. The development of next-generation sequencing (NGS) technologies has considerably improved our understanding of transcriptome complexity of Carex L. and provided a valuable genetic reference. However, the current transcriptome is not satisfactory mainly because of the enormous difficulty in obtaining full-length transcripts. Results In this study, we employed PacBio single-molecule long-read sequencing (SMRT) technology for whole-transcriptome profiling in Carex breviculmis. We generated 60,353 high-confidence non-redundant transcripts with an average length of 2302-bp. A total of 3588 alternative splicing events, and 1273 long non-coding RNAs were identified. Furthermore, 40,347 complete coding sequences were predicted, providing an informative reference transcriptome. In addition, the transcriptional regulation mechanism of C. breviculmis in response to shade stress was further explored by mapping the NGS data to the reference transcriptome constructed by SMRT sequencing. Conclusions This study provided a full-length reference transcriptome of C. breviculmis using the SMRT sequencing method for the first time. The transcriptome atlas obtained will not only facilitate future functional genomics studies but also pave the way for further selective and genic engineering breeding projects for C. breviculmis.

The bioenergy crop switchgrass (Panicum virgatum L.) has been recognized as friendly to the soil of cultivated land depending on the previous land use types and management practices. However, the effects of switchgrass establishment on soil properties at a broader depth when it is harvested annually without any fertilization in northern China largely remain unknown. To explore the impacts of unfertilized switchgrass on soil physical and chemical properties, 0–100 cm soil samples were collected from 7-year cropland-to-switchgrass conversion and the bare land (control). The results showed that switchgrass establishment increased soil total and capillary porosity, CFU numbers of the microbial communities (fungi, bacteria, and actinomycetes), contents of microbial biomass (carbon, nitrogen, and phosphorus), and water-soluble organic carbon, and decreased soil bulk density, mostly at 0–60 cm depths, compared to the control values. Notably, the annual harvest of switchgrass insignificantly increased soil total and available nitrogen contents and slightly reduced available phosphorus and potassium contents. In conclusion, long-term cropland conversion to unfertilized switchgrass could ameliorate soil properties and does not cause soil depletion. The output of this study could inspire governments and farmers to make large-scale use of switchgrass in the ecological restoration of abandoned cropland in north China.

Buffalograss [Buchloe dactyloides (Nutt.) Engelm.] has become the most widely cultivated warm-season turfgrass in northern China because of its low-maintenance requirements. Nitrogen (N) can be applied to plants in a range of formulations. However, preference of nitrogen uptake and the effects of N form on plant growth and nitrogen accumulation has not been established in buffalograss. In this study, we evaluated the effects of different inorganic nitrogen forms (NO3−-N, NH4+-N, and NO3−-N: NH4+-N = 1:1) on growth and nitrogen accumulation in buffalograss seedlings. Results showed that supply of three N forms significantly increased buffalograss seedlings growth, biomass, and N contents of all plant organs compared with the seedlings receiving free nitrogen. Plants achieved better growth performance when they received nitrate as the sole N source, which stimulated stolon growth and increased the biomass of ramets, spacers, and aboveground and total plant biomass, and also allocated more biomass to ramets and more N to spacers. Meanwhile, those plants supplied with the treatment +NH4NO3 displayed a significantly greater N content in the ramet, 15N abundance, and 15N accumulation amount in all organs. These data suggest NO3−-N supplied either singly or in mixture increased vegetative propagation and thus facilitates buffalograss establishment. However, applications of ammonium caused detrimental effects on buffalograss seedlings growth, but +NO3− could alleviate NH4+-induced morphological disorders. Thus, recommendations to increase vegetative propagation and biomass accumulation in buffalograss seedlings should consider increasing NO3−-N in a fertility program and avoiding applications of nitrogen as NH4+-N.

Forest ecosystems play an important role in regulating global temperatures through their capability to take up and store CO _2 from the atmosphere, but the magnitude and sustainability of this carbon ( C ) sink is critically dependent on the availability of nutrients, particularly nitrogen ( N ). However, the extent to which the absolute amount of N or modifications in plant and soil C:N ratios controls long-term forest carbon sequestration (C _seq ) remains uncertain. To assess this, we analyzed the results of 135 global field studies that investigated the dynamics of C and N availability during forest succession. The results showed that the accumulation of C and N in plant (including above- and below-ground vegetation) and litter pools decreased with forest age and approached an equilibrium value in the latter stages of stand development. Plant and litter C:N ratios increased during the first 10–20 years and remained relatively stable thereafter. The analysis further showed that the relative importance of a change in the total amount of N or modifications in the C:N ratio, to increases in C _seq , varied with forest age. Whilst the relative importance of a change in the total amount of N increased with forest age, the relative importance of a varied C:N stoichiometry decreased with forest age. Overall, a change in the total amount of N was the more important factor contributing to C storage during forest stand development and the C stored in vegetation dominated the total ecosystem C pool. These results show that ecosystem N availability is a key factor supporting long-term forest C _seq during forest succession. As most of the C is found in above-ground vegetation, this pool is particularly susceptible to abiotic or biotic factors and anthropogenically-related disturbances.

The partitioning of belowground biomass (BGB) to aboveground biomass (AGB) is commonly described as the root-to-shoot ratio (R/S). Although a number of studies have shown that biodiversity can influence AGB and BGB in grasslands at the local and global scale, the global-scale patterns reflecting how plant diversity affects R/S and the factors controlling such effects remain unclear. In this study, we explored the global patterns and associated drivers of biomass partitioning responding to plant diversity by conducting a meta-analysis of 333 observations from 30 studies in grasslands worldwide. Overall, plant diversity significantly increased AGB, BGB, and total biomass, whereas significantly decreased R/S. The effects of plant diversity on biomass partitioning varied with experimental types. The effect size for AGB and BGB in the field was larger than in greenhouse experiments, but the effect size for R/S did not significantly differ between field and greenhouse experiments. Moreover, there was no significant relationship between R/S and species richness and experimental duration in greenhouse experiments. However, the effect size for AGB, BGB, and R/S increased logarithmically with species richness and experimental duration in the field experiments. Specifically, the effect size for R/S in the field experiments switched from negative to neutral as the species richness and experimental duration increased. Furthermore, the effect size for R/S was positively correlated with complementary effects of BGB, and it increased logarithmically with mean annual temperature (MAT) and precipitation. Structural equation models showed that species richness, experimental duration, and MAT impact R/S indirectly by changing the BGB. Overall, our findings suggest that plant mixtures invest less in BGB than monocultures, and highlight that low investment in BGB will disappear gradually over time as species richness increases.

Growing evidence indicates that some grass species are more tolerant to various abiotic and biotic stresses than many crops. Zinc finger proteins play important roles in plant abiotic and biotic stresses. Although genes coding for these proteins have been cloned and identified in various plants, their function and underlying transcriptional mechanisms in the halophyte Zoysia japonica are barely known. In the present study, ZjZFN1 was isolated from Z. japonica using RACE method. Quantitative real time PCR results revealed that the expression of ZjZFN1 was much higher in leaf than in root and stem tissues, and induced by salt, cold or ABA treatment. The subcellular localization assay demonstrated that ZjZFN1 was localized to the nucleus. Expression of the ZjZFN1 in Arabidopsis thaliana improved seed germination and enhanced plant adaption to salinity stress with improved percentage of green cotyledons and growth status under salinity stress. Physiological and transcriptional analyses suggested that ZjZFN1 might, at least in part, influence reactive oxygen species accumulation and regulate the transcription of salinity responsive genes. Furthermore, RNA-sequencing analysis of ZjZFN1-overexpressing plants revealed that ZjZFN1 may serve as a transcriptional activator in the regulation of stress responsive pathways, including phenylalanine metabolism, α-linolenic acid metabolism and phenylpropanoid biosynthesis pathways. Taken together, these results provide evidence that ZjZFN1 is a potential key player in plants’ tolerance to salt stress, and it could be a valuable gene in Z. japonica breeding projects.

Switchgrass is a photoperiod-sensitive energy grass suitable for growing in the marginal lands of China. We explored the effects of extended photoperiods of low-irradiance light (7 μmol·m-2·s-1, no effective photosynthesis) on the growth, the biomass dry weight, the biomass allocation, and, especially, the stem digestibility and cell wall characteristics of switchgrass. Two extended photoperiods (i.e., 18 and 24 h) were applied over Alamo. Extended light exposure (18 and 24 h) resulted in delayed heading and higher dry weights of vegetative organs (by 32.87 and 35.94%, respectively) at the expense of reducing the amount of sexual organs (by 40.05 and 50.87%, respectively). Compared to the control group (i.e., natural photoperiod), the yield of hexoses (% dry matter) in the stems after a direct enzymatic hydrolysis (DEH) treatment significantly increased (by 44.02 and 46.10%) for those groups irradiated during 18 and 24 h, respectively. Moreover, the yield of hexoses obtained via enzymatic hydrolysis increased after both basic (1% NaOH) and acid (1% H2SO4) pretreatments for the groups irradiated during 18 and 24 h. Additionally, low-irradiance light extension (LILE) significantly increased the content of non-structural carbohydrates (NSCs) while notably reducing the lignin content and the syringyl to guaiacyl (S/G) ratio. These structural changes were in part responsible for the observed improved stem digestibility. Remarkably, LILE significantly decreased the cellulose crystallinity index (CrI) of switchgrass by significantly increasing both the arabinose substitution degree in xylan and the content of ammonium oxalate-extractable uronic acids, both favoring cellulose digestibility. Despite this LILE technology is not applied to the cultivation of switchgrass on a large scale yet, we believe that the present work is important in that it reveals important relationships between extended day length irradiations and biomass production and quality. Additionally, this study paves the way for improving biomass production and digestibility via genetic modification of day length sensitive transcription factors or key structural genes in switchgrass leaves.

Switchgrass (Panicum virgatum L.) is a warm-season rhizomatous perennial grass that can tolerate diverse abiotic stresses while yielding relatively high biomass, and is considered a leading biofuel feedstock for marginal lands. Nitrogen (N) is crucial for the growth and development of switchgrass, and its tolerance to low N supply and high N use efficiency are very important for its production under poor conditions. The large-scale planting of switchgrass on marginal lands could be an effective approach to solving the problem of feedstock supply for biomass energy. This study used a hydroponic experiment to evaluate the effect of N deficiency on switchgrass seedlings. Three N treatments (0, 0.15, and 1.50 mmol L− 1 Hoagland's solution) and six cultivars were used, three of each ecotype (upland and lowland). The results showed that biomass, leaf area, root surface area, net photosynthesis, and total chlorophyll content significantly decreased under low N treatments compared with those in full strength Hoagland's nutrient solution. However, once established, all plants survived extreme N stress (0 mmol L− 1) and, to some extent, were productive. Cultivar Kanlow performed best of the six cultivars under stress. Significant interactions between stress treatment and cultivars showed that breeding for cultivars with high yield and superior performance under N deficiency is warranted. The lowland outperformed the upland ecotypes under stress, suggesting that lowland cultivars may survive and be productive under a wider range of stress conditions. However, given the better adaptability of lowland ecotypes to hydroponic cultivation, further study is needed.

In this study, tassels of Cave-in-Rock (upland) and Alamo (lowland) were removed at or near tassel emergence to explore its effects on biomass production and quality. Tassel-removed (TR) Cave-in-Rock and Alamo both exhibited a significant (P