N6-methyladenosine analysis unveils key mechanisms underlying long-term salt stress tolerance in switchgrass (Panicum virgatum).

N6-methyladenosine (m6A) RNA modification is critical for plant growth, development, and environmental stress response. While short-term stress impacts on m6A are well-documented, the consequences of prolonged stress remain underexplored. This study conducts a thorough transcriptome-wide analysis of m6A modifications following 28-day exposure to 200 mM NaCl. We detected 11,149 differentially expressed genes (DEGs) and 12,936 differentially methylated m6A peaks, along with a global decrease in m6A levels. Notably, about 62% of m6A-modified DEGs, including demethylase genes like PvALKBH6_N, PvALKBH9_K, and PvALKBH10_N, showed increased expression and reduced m6A peaks, suggesting that decreased m6A methylation may enhance gene expression under salt stress. Consistent expression and methylation patterns were observed in key genes related to ion homeostasis (e.g., H+-ATPase 1, High-affinity K+transporter 5), antioxidant defense (Catalase 1/2, Copper/zinc superoxide dismutase 2, Glutathione synthetase 1), and osmotic regulation (delta 1-pyrroline-5-carboxylate synthase 2, Pyrroline-5-carboxylate reductase). These findings provide insights into the adaptive mechanisms of switchgrass under long-term salt stress and highlight the potential of regulating m6A modifications as a novel approach for crop breeding strategies focused on stress resistance.

Nanosilica enhances morphogenic and chemical parameters of Megathyrsus maximus grass under conditions of phosphorus deficiency and excess stress in different soils.

Phosphorus (P) imbalances are a recurring issue in cultivated soils with pastures across diverse regions. In addition to P deficiency, the prevalence of excess P in soil has escalated, resulting in damage to pasture yield. In response to this reality, there is a need for well-considered strategies, such as the application of silicon (Si), a known element for alleviating plant stress. However, the influence of Si on the morphogenetic and chemical attributes of forage grasses grown in various soils remains uncertain. Consequently, this study aimed to assess the impact of P deficiency and excess on morphogenetic and chemical parameters, as well as digestibility, in Zuri guinea grass cultivated in Oxisol and Entisol soils. It also sought to determine whether fertigation with nanosilica could mitigate the detrimental effects of these nutritional stresses. Results revealed that P deficiency led to a reduction in tiller numbers and grass protein content, along with an increase in lignin content. Conversely, P excess resulted in higher proportions of dead material and lignin, a reduced mass leaf: stem ratio in plants, and a decrease in dry matter (DM) yield. Fertigation with Si improved tillering and protein content in deficient plants. In the case of P excess, Si reduced tiller mortality and lignin content, increased the mass leaf:stem ratio, and enhanced DM yield. This approach also increased yields in plants with sufficient P levels without affecting grass digestibility. Thus, Si utilization holds promise for enhancing the growth and chemical characteristics of forage grasses under P stress and optimizing yield in well-nourished, adapted plants, promoting more sustainable pasture yields.

A generalist-specialist trade-off between switchgrass cytotypes impacts climate adaptation and geographic range.

Polyploidy results from whole-genome duplication and is a unique form of heritable variation with pronounced evolutionary implications. Different ploidy levels, or cytotypes, can exist within a single species, and such systems provide an opportunity to assess how ploidy variation alters phenotypic novelty, adaptability, and fitness, which can, in turn, drive the development of unique ecological niches that promote the coexistence of multiple cytotypes. Switchgrass, Panicum virgatum, is a widespread, perennial C4 grass in North America with multiple naturally occurring cytotypes, primarily tetraploids (4×) and octoploids (8×). Using a combination of genomic, quantitative genetic, landscape, and niche modeling approaches, we detect divergent levels of genetic admixture, evidence of niche differentiation, and differential environmental sensitivity between switchgrass cytotypes. Taken together, these findings support a generalist (8×)­specialist (4×) trade-off. Our results indicate that the 8× represent a unique combination of genetic variation that has allowed the expansion of switchgrass' ecological niche and thus putatively represents a valuable breeding resource.

Cultivar sensitivity of broomcorn millet (Panicum miliaceum L.) to nitrogen availability is associated with differences in photosynthetic physiology and nitrogen uptake.

In order to explore the influences of low nitrogen (N) fertilizer on the growth performances of two broomcorn millet (Panicum miliaceum L.) cultivars with different N tolerances, the field experiment was carried out with a low-N-tolerant cultivar (BM 184) and a low-N-sensitive cultivar (BM 230) under three N levels (0, 75 and 150 kg N ha-1) in the Loess Plateau, China. 150 kg N ha-1 was conventional N application rate and considered as the control. Compared to typical N supply, low N fertilizer significantly weakened the photosynthetic capacity by increasing the light transmission ratio and decreasing leaf area index, resulting in reduced biomass accumulation. BM 184 held the longer duration of the biomass increase phase and larger relative growth rate than BM 230 as well as higher photosynthetic parameters (i.e., relative chlorophyll content, net photosynthetic rate, and transpiration rate) did under low N treatments. Such optimized physiological characteristics contributed to more effective N uptake and transportation from the stems, leaves, and sheaths to grains in the BM 184. Furthermore, compared with BM 230, BM 184 had higher rhizosphere soil fertility and soil enzyme activity under low N conditions; consequently, combined with the physiological characteristics for aboveground and soil nutrient status for belowground, higher productivity was obtained in BM 184 than that in BM 230 over the two years study. Overall, our results demonstrated that low-N-tolerant cultivar achieved reduced N fertilizer input with increased efficiency by optimizing growth performances in semi-arid cultivation areas.

Evaluation of the physiological potential of Panicum maximum seeds by multivariate analysis

The aim of this work was to identify efficient vigor tests for differentiating the seed lots, forecasting seedling emergence in the field and assessing the physiological quality of Panicum maximum seeds. 12 seed lots from the cultivar Tanzania and 11 seed lots from the cultivar Massai were evaluated for water content, germination, first count and germination speed index, emergence and first emergence count of seedlings in sand, root length and shoot length, analysis of SVIS® images (seedling length, vigor and uniformity index) and seedling emergence in the field. The work was conducted in a completely randomized design for tests performed in the laboratory and in randomized blocks for tests in the field. The data were subjected to analysis of variance and the means compared by Scott Knott's test at 5% probability and statistical multivariate clustering analysis and principal components analysis. The shoot and root length tests are efficient for the evaluation of the physiological potential of P. maximum cv. Massai, while the seedling length, vigor index and growth uniformity index tests using image analysis, seedling emergence in sand and first seedling emergence count in sand are efficient in assessing the physiological potential of seeds of P. maximum cv. Tanzania, and providing information similar to that of seedling emergence in the field.

Drought stress and plant ecotype drive microbiome recruitment in switchgrass rhizosheath.

The rhizosheath, a layer of soil grains that adheres firmly to roots, is beneficial for plant growth and adaptation to drought environments. Switchgrass is a perennial C4 grass which can form contact rhizosheath under drought conditions. In this study, we characterized the microbiomes of four different rhizocompartments of two switchgrass ecotypes (Alamo and Kanlow) grown under drought or well-watered conditions via 16S ribosomal RNA amplicon sequencing. These four rhizocompartments, the bulk soil, rhizosheath soil, rhizoplane, and root endosphere, harbored both distinct and overlapping microbial communities. The root compartments (rhizoplane and root endosphere) displayed low-complexity communities dominated by Proteobacteria and Firmicutes. Compared to bulk soil, Cyanobacteria and Bacteroidetes were selectively enriched, while Proteobacteria and Firmicutes were selectively depleted, in rhizosheath soil. Taxa from Proteobacteria or Firmicutes were specifically selected in Alamo or Kanlow rhizosheath soil. Following drought stress, Citrobacter and Acinetobacter were further enriched in rhizosheath soil, suggesting that rhizosheath microbiome assembly is driven by drought stress. Additionally, the ecotype-specific recruitment of rhizosheath microbiome reveals their differences in drought stress responses. Collectively, these results shed light on rhizosheath microbiome recruitment in switchgrass and lay the foundation for the improvement of drought tolerance in switchgrass by regulating the rhizosheath microbiome.

Transcriptional, metabolic, physiological and developmental responses of switchgrass to phosphorus limitation.

Knowing how switchgrass (Panicum virgatum L.) responds and adapts to phosphorus (P)-limitation will aid efforts to optimize P acquisition and use in this species for sustainable biomass production. This integrative study investigated the impacts of mild, moderate, and severe P-stress on genome transcription and whole-plant metabolism, physiology and development in switchgrass. P-limitation reduced overall plant growth, increased root/shoot ratio, increased root branching at moderate P-stress, and decreased root diameter with increased density and length of root hairs at severe P-stress. RNA-seq analysis revealed thousands of genes that were differentially expressed under moderate and severe P-stress in roots and/or shoots compared to P-replete plants, with many stress-induced genes involved in transcriptional and other forms of regulation, primary and secondary metabolism, transport, and other processes involved in P-acquisition and homeostasis. Amongst the latter were multiple miRNA399 genes and putative targets of these. Metabolite profiling showed that levels of most sugars and sugar alcohols decreased with increasing P stress, while organic and amino acids increased under mild and moderate P-stress in shoots and roots, although this trend reversed under severe P-stress, especially in shoots.

QTL mapping of winter dormancy and associated traits in two switchgrass pseudo-F1 populations: lowland x lowland and lowland x upland.

BACKGROUND: Switchgrass (Panicum virgatum) undergoes winter dormancy by sensing photoperiod and temperature changes. It transitions to winter dormancy in early fall following at the end of reproduction and exits dormancy in the spring. The duration of the growing season affects the accumulation of biomass and yield. In this study, we conducted QTL mapping of winter dormancy measured by fall regrowth height (FRH) and normalized difference vegetation index (NDVI), spring emergence (SE), and flowering date (FD) in two bi-parental pseudo-F1 populations derived from crosses between the lowland AP13 with the lowland B6 (AB) with 285 progenies, and the lowland B6 with the upland VS16 (BV) with 227 progenies. RESULTS: We identified 18 QTLs for FRH, 18 QTLs for NDVI, 21 QTLs for SE, and 30 QTLs for FD. The percent variance explained by these QTLs ranged between 4.21-23.27% for FRH, 4.47-24.06% for NDVI, 4.35-32.77% for SE, and 4.61-29.74% for FD. A higher number of QTL was discovered in the BV population, suggesting more variants in the lowland x upland population contributing to the expression of seasonal dormancy underlying traits. We identified 9 regions of colocalized QTL with possible pleiotropic gene action. The positive correlation between FRH or NDVI with dry biomass weight suggests that winter dormancy duration could affect switchgrass biomass yield. The medium to high heritability levels of FRH (0.55-0.64 H2) and NDVI (0.30-0.61 H2) implies the possibility of using the traits for indirect selection for biomass yield. CONCLUSION: Markers found within the significant QTL interval can serve as genomic resources for breeding non-dormant and semi-dormant switchgrass cultivars for the southern regions, where growers can benefit from the longer production season.

ADP-ribosylation factors improve biomass yield and salinity tolerance in transgenic switchgrass (Panicum virgatum L.).

KEY MESSAGE: PvArf regulate proline biosynthesis by physically interacting with PvP5CS1 to improve salt tolerance in switchgrass. The genetic factors that contribute to stress resiliency are yet to be determined. Here, we identified three ADP-ribosylation factors, PvArf1, PvArf-B1C, and PvArf-related, which contribute to salinity tolerance in transgenic switchgrass (Panicum virgatum L.). Switchgrass overexpressing each of these genes produced approximately twofold more biomass than wild type (WT) under normal growth conditions. Transgenic plants accumulated modestly higher levels of proline under normal conditions, but this level was significantly increased under salt stress providing better protection to transgenic plants compared to WT. We found that PvArf genes induce proline biosynthesis genes under salt stress to positively regulate proline accumulation, and further demonstrated that PvArf physically interact with PvP5CS1. Moreover, the transcript levels of two key ROS-scavenging enzyme genes were significantly increased in the transgenic plants compared to WT, leading to reduced H2O2 accumulation under salt stress conditions. PvArf genes also protect cells against stress-induced changes in Na+ and K+ ion concentrations. Our findings uncover that ADP-ribosylation factors are key determinants of biomass yield in switchgrass, and play pivotal roles in salinity tolerance by regulating genes involved in proline biosynthesis.

Low-nitrogen tolerance comprehensive evaluation and physiological response to nitrogen stress in broomcorn millet (Panicum miliaceum L.) seedling.

Developing the new crop varieties with high productivity under low nitrogen (N) input is an important access to facilitate modern agricultural sustainability. In the present study, 20 broomcorn millet (Panicum miliaceum L.) varieties were characterized for their morphological and nutrient parameters to different low N levels in seedling. The results showed that 0.25 mM NH4NO3 was the standard concentration for the evaluation and identification of low-N tolerance. Through pearson's correlation analysis, principal component analysis, and subordinate function analysis, the tolerance of 20 varieties under N stress was evaluated and plant height, root length, shoot biomass, and shoot and root N content were considered as the evaluation system of low-N tolerance. Although leaves photosynthetic capacities and activities of N metabolism related enzymes showed the decreasing tendency to N stress, low-N tolerant varieties had higher activities in both leaves and roots as compared to low-N sensitive varieties. The work provides a reliable and comprehensive method for evaluating low-N tolerance in broomcorn millet and our data elucidate possible physiological adaptive mechanisms by which broomcorn millet tolerates N stress.

Cyclic electron flow and light partitioning between the two photosystems in leaves of plants with different functional types.

Cyclic electron flow (CEF) around photosystem I (PSI) is essential for generating additional ATP and enhancing efficient photosynthesis. Accurate estimation of CEF requires knowledge of the fractions of absorbed light by PSI (fI) and PSII (fII), which are only known for a few model species such as spinach. No measures of fI are available for C4 grasses under different irradiances. We developed a new method to estimate (1) fII in vivo by concurrently measuring linear electron flux through both photosystems [Formula: see text] in leaf using membrane inlet mass spectrometry (MIMS) and total electron flux through PSII (ETR2) using chlorophyll fluorescence by a Dual-PAM at low light and (2) CEF as ETR1-[Formula: see text]. For a C3 grass, fI was 0.5 and 0.4 under control (high light) and shade conditions, respectively. C4 species belonging to NADP-ME and NAD-ME subtypes had fI of 0.6 and PCK subtype had 0.5 under control. All shade-grown C4 species had fI of 0.6 except for NADP-ME grass which had 0.7. It was also observed that fI ranged between 0.3 and 0.5 for gymnosperm, liverwort and fern species. CEF increased with irradiance and was induced at lower irradiances in C4 grasses and fern relative to other species. CEF was greater in shade-grown plants relative to control plants except for C4 NADP-ME species. Our study reveals a range of CEF and fI values in different plant functional groups. This variation must be taken into account for improved photosynthetic calculations and modelling.

Comparative analysis of proso millet (Panicum miliaceum L.) leaf transcriptomes for insight into drought tolerance mechanisms.

BACKGROUND: Drought stress is a major abiotic stress that causes huge losses in agricultural production. Proso millet (Panicum miliaceum L.) can efficiently adapt to drought stress and provides important information and gene resources to improve drought tolerance. However, its complex drought-responsive mechanisms remain unclear. RESULTS: Among 37 core Chinese proso millet cultivars, Jinshu 6 (JS6) was selected as the drought-sensitive test material, whereas Neimi 5 (NM5) was selected as the drought-tolerant test material under PEG-induced water stress. After sequencing, 1695 differentially expressed genes (DEGs) were observed in JS6 and NM5 without PEG-induced water stress (JS6CK and NM5CK). A total of 833 and 2166 DEGs were found in the two cultivars under simulated drought by using 20% PEG-6000 for 6 (JS6T6 and NM5T6) and 24 h (JS6T24 and NM5T24), respectively. The DEGs in JS6T6 and JS6T24 treatments were approximately 0.298- and 0.754-fold higher than those in NM5T6 and NM5T24, respectively. Compared with the respective controls, more DEGs were found in T6 treatments than in T24 treatments. A delay in the transcriptional responses of the ROS scavenging system to simulated drought treatment and relatively easy recovery of the expression of photosynthesis-associated genes were observed in NM5. Compared with JS6, different regulation strategies were observed in the jasmonic acid (JA) signal transduction pathway of NM5. CONCLUSION: Under PEG-induced water stress, NM5 maintained highly stable gene expression levels. Compared with drought-sensitive cultivars, the different regulation strategies in the JA signal transduction pathway in drought-tolerant cultivars may be one of the driving forces underlying drought stress tolerance.

Ozone phytotoxicity to Panicum maximum and Cenchrus ciliaris at Indo-Gangetic plains: an assessment of antioxidative defense and growth responses.

Two common tropical grassland species, Panicum maximum Jacq. (Guinea grass) and Cenchrus ciliaris (Buffel grass) of Indo-Gangetic plains were assessed for their responses under future level of O3 (ambient +30 ppb) using open top chambers. Plants were assessed for foliar injuries, pigments, growth, biomass accumulation, histochemical localization of reactive oxygen species (ROS), antioxidant defense system and ROS scavenging activities at two stages. Foliar injuries were noticed at an early stage in P. maximum compared to C. ciliaris. Significant reductions were observed in total chlorophyll, growth and total biomass in both species. Significant increases in contents of melondialdehyde and ascorbic acid in P. maximum while total phenolics and thiols in C. ciliaris were found. Histochemical analysis showed more production of superoxide radicals and hydrogen peroxide in leaf tissues of P. maximum compared to C. ciliaris. It can be concluded that higher level of primary antioxidants (total phenolics and thiols) along with superoxide dismutase and ascorbate peroxidase scavenged O3 effectively in C. ciliaris causing less reduction of biomass which is used as a feed for cattles. In P. maximum, more photosynthates were allocated for defense, leading to higher reduction in total biomass compared to C. ciliaris. The leaf area ratio was higher in P. maximum compared to C. ciliaris under elevated O3. The study further suggests higher susceptibility of P. maximum compared to C. ciliaris under future level of O3 exposure.

Potassium affects the phytoextraction potential of Tanzania guinea grass under cadmium stress.

The supply of potassium (K) is a strategy to increase the tolerance of plants exposed to Cd toxicity. The aim of this study was to verify the influence of K on the growth and potential of Tanzania guinea grass (Panicum maximum Jacq. cv. Tanzania (syn. Megathyrsus maximus (Jacq.) B.K. Simon & S.W.L. Jacobs)) for Cd phytoextraction as well as to evaluate nutritional attributes of this grass under conditions of Cd stress. The experiment was conducted in a randomized complete block design, using a 3 × 4 factorial arrangement, with three replications. Three rates of K (0.4, 6.0, and 11.6 mmol L-1) were combined with four rates of Cd (0.0, 0.5, 1.0, and 1.5 mmol L-1) in nutrient solution. Two plant growth periods were evaluated. The increase in K supply to plants exposed to Cd rates of up to 1.0 mmol L-1 caused increase in morphogenic and production attributes, as well as reduction in tiller mortality rate, in the second growth period. K concentrations (in both harvests) increased, while calcium and magnesium concentrations in the second harvest decreased with increasing Cd rates. The high availability of Cd (1.5 mmol L-1) in the nutrient solution caused decrease in relative chlorophyll index (RCI) in both harvests. The high supply of K to plants exposed to Cd resulted in high shoot dry mass production, reducing Cd concentration in the photosynthetic tissues (which means great tolerance of the plant) and increasing the accumulation of this metal in the shoots that can be harvested. Therefore, K increases the Cd phytoextraction capacity of Tanzania guinea grass.

Overexpression of the Lolium perenne L. delta1-pyrroline 5-carboxylate synthase (LpP5CS) gene results in morphological alterations and salinity tolerance in switchgrass (Panicum virgatum L.).

In plants, Δ1-pyrroline- 5-carboxylate synthase (P5CS) is the rate-limiting enzyme in proline biosynthesis. In this study, we introduced the LpP5CS (Lolium perenne L.) gene into switchgrass by Agrobacterium-mediated transformation. The transgenic lines (TG) were classified into two groups based on their phenotypes and proline levels. The group I lines (TG4 and TG6) had relatively high proline levels and improved biomass yield. The group II lines (TG1 and TG2) showed low proline levels, severely delayed flowering, stunted growth and reduced biomass yield. Additionally, we used RNA-seq analysis to detect the most significant molecular changes, and we analyzed differentially expressed genes, such as flowering-related and CYP450 family genes. Moreover, the biomass yield, physiological parameters, and expression levels of reactive oxygen species scavenger-related genes under salt stress all indicated that the group I plants exhibited significantly increased salt tolerance compared with that of the control plants, in contrast to the group II plants. Thus, genetic improvement of switchgrass by overexpressing LpP5CS to increase proline levels is feasible for increasing plant stress tolerance.

Comparative metabolite profiling of two switchgrass ecotypes reveals differences in drought stress responses and rhizosheath weight.

MAIN CONCLUSION: Rhizosheath comprises soil that adheres firmly to roots. In this study, two ecotypes of switchgrass with different rhizosheath sizes after drought stress were analyzed which showed metabolic differences under drought conditions. The rhizosheath comprises soil that adheres firmly to roots by a combination of root hairs and mucilage and may aid in root growth under soil drying. The aim of this work is to reveal the potential metabolites involved in rhizosheath formation under drought stress conditions. Panicum virgatum L. (switchgrass), which belongs to the Poaceae family, is an important biofuel and fodder crop in drought areas. Five switchgrass ecotypes (cv. Alamo, cv. Blackwake, cv. Summer, cv. Cave-in-Rock and cv. Kanlow) have a broad range of rhizosheath weight under drought conditions. For two selected ecotypes with contrast rhizosheath weight (cv. Alamo and cv. Kanlow), root hair length and density, lateral root number, root morphological parameters were measured, and real-time qRT-PCR was performed. Gas chromatography mass spectrophotometry (GC-MS) was used to determine the primary metabolites in the shoots and roots of selected ecotypes under drought stress conditions. The change trends of root hair length and density, lateral root number and related gene expression were consistent with rhizosheath weight in Alamo and Kanlow under drought and watered conditions. For root morphological parameters, Alamo grew deeper than Kanlow, while Kanlow exhibited higher values for other parameters. In this study, the levels of amino acids, sugars and organic acids were significantly changed in response to drought stress in two switchgrass ecotypes. Several metabolites including amino acids (arginine, isoleucine, methionine and cysteine) and sugars (kestose, raffinose, fructose, fucose, sorbose and xylose) in the large soil-sheathed roots of Alamo and Kanlow were significantly increased compared to small or no soil-sheathed roots of Alamo and Kanlow. Difference in rhizosheath size is reflected in the plant internal metabolites under drought stress conditions. Additionally, our results highlight the importance of using metabolite profiling and provide a better understanding of rhizosheath formation at the cellular level.

QTL × environment interactions underlie adaptive divergence in switchgrass across a large latitudinal gradient.

Local adaptation is the process by which natural selection drives adaptive phenotypic divergence across environmental gradients. Theory suggests that local adaptation results from genetic trade-offs at individual genetic loci, where adaptation to one set of environmental conditions results in a cost to fitness in alternative environments. However, the degree to which there are costs associated with local adaptation is poorly understood because most of these experiments rely on two-site reciprocal transplant experiments. Here, we quantify the benefits and costs of locally adaptive loci across 17° of latitude in a four-grandparent outbred mapping population in outcrossing switchgrass (Panicum virgatum L.), an emerging biofuel crop and dominant tallgrass species. We conducted quantitative trait locus (QTL) mapping across 10 sites, ranging from Texas to South Dakota. This analysis revealed that beneficial biomass (fitness) QTL generally incur minimal costs when transplanted to other field sites distributed over a large climatic gradient over the 2 y of our study. Therefore, locally advantageous alleles could potentially be combined across multiple loci through breeding to create high-yielding regionally adapted cultivars.

Short-term warming and water stress affect Panicum maximum Jacq. stoichiometric homeostasis and biomass production.

Climate changes affect the growth of forage species. However, information regarding the effects of global climate change on the stoichiometry of tropical pastures is lacking, especially under field conditions. Such information is crucial to understand how temperature conditions and water availability states are likely to affect the stoichiometric homeostasis and biomass production of Panicum maximum, an important C4 tropical forage species, under future climate change scenarios. Thus, we, conducted a field experiment using a temperature free-air controlled enhancement system and evaluated the effects of two temperature conditions, ambient temperature and moderate warming (2 °C above ambient canopy temperature), and two levels of water availability, irrigated and non-irrigated, on the stoichiometric patterns of C:N:P and leaf biomass production. The experiment was conducted using a randomized complete block design in a factorial arrangement with four replications over 3 weeks. Our findings revealed that the N and P leaf concentration greatly decreased in water-stressed plants, which increased the C:N and C:P ratios, while warming increased the N:P ratio. Leaf biomass production was impaired by up to 16% under water stress and ambient temperature conditions, but the biomass production was improved by 20% under warming and irrigated conditions. Our findings showed that homeostatic instability under rainfed conditions resulted in decreased leaf biomass production. Therefore, we concluded that warming is only beneficial for plant growth (i.e., a high homeostatic capacity was maintained) under well-irrigated conditions.

Increasing atmospheric CO2 and canopy temperature induces anatomical and physiological changes in leaves of the C4 forage species Panicum maximum.

Changes in leaf anatomy and ultrastructure are associated with physiological performance in the context of plant adaptations to climate change. In this study, we investigated the isolated and combined effects of elevated atmospheric CO2 concentration ([CO2]) up to 600 µmol mol-1 (eC) and elevated temperature (eT) to 2°C more than the ambient canopy temperature on the ultrastructure, leaf anatomy, and physiology of Panicum maximum Jacq. grown under field conditions using combined free-air carbon dioxide enrichment (FACE) and temperature free-air controlled enhancement (T-FACE) systems. Plants grown under eC showed reduced stomatal density, stomatal index, stomatal conductance (gs), and leaf transpiration rate (E), increased soil-water content (SWC) conservation and adaxial epidermis thickness were also observed. The net photosynthesis rate (A) and intrinsic water-use efficiency (iWUE) were enhanced by 25% and 71%, respectively, with a concomitant increase in the size of starch grains in bundle sheath cells. Under air warming, we observed an increase in the thickness of the adaxial cuticle and a decrease in the leaf thickness, size of vascular bundles and bulliform cells, and starch content. Under eCeT, air warming offset the eC effects on SWC and E, and no interactions between [CO2] and temperature for leaf anatomy were observed. Elevated [CO2] exerted more effects on external characteristics, such as the epidermis anatomy and leaf gas exchange, while air warming affected mainly the leaf structure. We conclude that differential anatomical and physiological adjustments contributed to the acclimation of P. maximum growing under elevated [CO2] and air warming, improving the leaf biomass production under these conditions.

SPL7 and SPL8 represent a novel flowering regulation mechanism in switchgrass.

The aging pathway in flowering regulation is controlled mainly by microRNA156 (miR156). Studies in Arabidopsis thaliana reveal that nine miR156-targeted SQUAMOSA PROMOTER BINDING-LIKE (SPL) genes are involved in the control of flowering. However, the roles of SPLs in flowering remain elusive in grasses. Inflorescence development in switchgrass was characterized using scanning electron microscopy (SEM). Microarray, quantitative reverse transcription polymerase chain reaction (qRT-PCR), chromatin immunoprecipitation (ChIP)-PCR and EMSA were used to identify regulators of phase transition and flowering. Gene function was characterized by downregulation and overexpression of the target genes. Overexpression of SPL7 and SPL8 promotes flowering, whereas downregulation of individual genes moderately delays flowering. Simultaneous downregulation of SPL7/SPL8 results in extremely delayed or nonflowering plants. Furthermore, downregulation of both genes leads to a vegetative-to-reproductive reversion in the inflorescence, a phenomenon that has not been reported in any other grasses. Detailed analyses demonstrate that SPL7 and SPL8 induce phase transition and flowering in grasses by directly upregulating SEPALLATA3 (SEP3) and MADS32. Thus, the SPL7/8 pathway represents a novel regulatory mechanism in grasses that is largely different from that in Arabidopsis. Additionally, genetic modification of SPL7 and SPL8 results in much taller plants with significantly increased biomass yield and sugar release.