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At least 199 records · Page 11

Altering the substitution and cross‐linking of glucuronoarabinoxylans affects cell wall architecture in Brachypodium distachyon

Summary The Poaceae family of plants provides cereal crops that are critical for human and animal nutrition, and also, they are an important source of biomass. Interacting plant cell wall components give rise to recalcitrance to digestion; thus, understanding the wall molecular architecture is important to improve biomass properties. Xylan is the main hemicellulose in grass cell walls. Recently, we reported structural variation in grass xylans, suggesting functional specialisation and distinct interactions with cellulose and lignin. Here, we investigated the functions of these xylans by perturbing the biosynthesis of specific xylan types. We generated CRISPR/Cas9 knockout mutants in Brachypodium distachyon XAX1 and GUX2 genes involved in xylan substitution. Using carbohydrate gel electrophoresis, we identified biochemical changes in different xylan types. Saccharification, cryo‐SEM, subcritical water extraction and ssNMR were used to study wall architecture. Bd XAX1A and Bd GUX2 enzymes modify different types of grass xylan. Brachypodium mutant walls are likely more porous, suggesting the xylan substitutions directed by both Bd XAX1A and GUX2 enzymes influence xylan‐xylan and/or xylan–lignin interactions. Since xylan substitutions influence wall architecture and digestibility, our findings open new avenues to improve cereals for food and to use grass biomass for feed and the production of bioenergy and biomaterials.

59 BASIC BIOLOGICAL SCIENCES↗

Simultaneous suppression of lignin, tricin and wall‐bound phenolic biosynthesis via the expression of monolignol 4‐ O ‐methyltransferases in rice

Summary Grass lignocelluloses feature complex compositions and structures. In addition to the presence of conventional lignin units from monolignols, acylated monolignols and flavonoid tricin also incorporate into lignin polymer; moreover, hydroxycinnamates, particularly ferulate, cross‐link arabinoxylan chains with each other and/or with lignin polymers. These structural complexities make grass lignocellulosics difficult to optimize for effective agro‐industrial applications. In the present study, we assess the applications of two engineered monolignol 4‐ O ‐methyltransferases (MOMTs) in modifying rice lignocellulosic properties. Two MOMTs confer regiospecific para ‐methylation of monolignols but with different catalytic preferences. The expression of MOMTs in rice resulted in differential but drastic suppression of lignin deposition, showing more than 50% decrease in guaiacyl lignin and up to an 90% reduction in syringyl lignin in transgenic lines. Moreover, the levels of arabinoxylan‐bound ferulate were reduced by up to 50%, and the levels of tricin in lignin fraction were also substantially reduced. Concomitantly, up to 11 μmol/g of the methanol‐extractable 4‐ O ‐methylated ferulic acid and 5–7 μmol/g 4‐ O ‐methylated sinapic acid were accumulated in MOMT transgenic lines. Both MOMTs in vitro displayed discernible substrate promiscuity towards a range of phenolics in addition to the dominant substrate monolignols, which partially explains their broad effects on grass phenolic biosynthesis. The cell wall structural and compositional changes resulted in up to 30% increase in saccharification yield of the de‐starched rice straw biomass after diluted acid‐pretreatment. These results demonstrate an effective strategy to tailor complex grass cell walls to generate improved cellulosic feedstocks for the fermentable sugar‐based production of biofuel and bio‐chemicals.

4-O-methylated↗

Metabolic Interactions between Brachypodium and Pseudomonas fluorescens under Controlled Iron-Limited Conditions

Plant iron (Fe) nutritional status has a significant impact on rhizosphere microbial communities. While this effect has been attributed to alterations in the composition of root exudates, the underlying mechanisms are not known. Here, we investigated the effect of Fe deficiency on the interactions between the grass Brachypodium distachyon and the common soil bacteria Pseudomonas fluorescens SBW25. Increased phytosiderophore production was the most significant response of Brachypodium root exudation to Fe deficiency. We observed upregulated expression of phytosiderophore biosynthesis genes but lower accumulated exudate concentrations in the presence of Pseudomonas, indicating that the bacteria degrade these phytosiderophores. Pseudomonas did not produce endogenous siderophores under the low Fe conditions. Rather, the bacterial transcriptome revealed upregulation of four genes in response to Fe deficiency that contain motifs associated with cellular uptake of small molecules. Collectively, these results suggest that rhizosphere Pseudomonas fluorescens may take up phytosiderophores produced by grasses in response to Fe deficiency, and we propose target genes that may be involved. Our findings provide insight into the molecular basis for the exchange of C, N, and Fe between plants and bacteria under Fe deficient conditions. These interactions may contribute to differences in the rhizosphere microbial community structure across soil pH regimes.

Boiteau, Rene M.↗

Data for Simultaneous Suppression of Lignin, Tricin and Wall-Bound Phenolic Biosynthesis via the Expression of Monolignol 4- O -Methyltransferases in Rice

Grass lignocelluloses feature complex compositions and structures. In addition to the presence of conventional lignin units from monolignols, acylated monolignols and flavonoid tricin also incorporate into lignin polymer; moreover, hydroxycinnamates, particularly ferulate, cross-link arabinoxylan chains with each other and/or with lignin polymers. These structural complexities make grass lignocellulosics difficult to optimize for effective agro-industrial applications. In the present study, we assess the applications of two engineered monolignol 4- O -methyltransferases (MOMTs) in modifying rice lignocellulosic properties. Two MOMTs confer regiospecific para-methylation of monolignols but with different catalytic preferences. The expression of MOMTs in rice resulted in differential but drastic suppression of lignin deposition, showing more than 50% decrease in guaiacyl lignin and up to an 90% reduction in syringyl lignin in transgenic lines. Moreover, the levels of arabinoxylan-bound ferulate were reduced by up to 50%, and the levels of tricin in lignin fraction were also substantially reduced. Concomitantly, up to 11 μmol/g of the methanol-extractable 4- O -methylated ferulic acid and 5–7 μmol/g 4- O -methylated sinapic acid were accumulated in MOMT transgenic lines. Both MOMTs in vitro displayed discernible substrate promiscuity towards a range of phenolics in addition to the dominant substrate monolignols, which partially explains their broad effects on grass phenolic biosynthesis. The cell wall structural and compositional changes resulted in up to 30% increase in saccharification yield of the de-starched rice straw biomass after diluted acid-pretreatment. These results demonstrate an effective strategy to tailor complex grass cell walls to generate improved cellulosic feedstocks for the fermentable sugar-based production of biofuel and bio-chemicals.

Biomass Analytics↗

A trait-based framework for linking microbial communities with carbon transformations under precipitation change

Droughts are common throughout the world. As the climate changes, droughts may become more frequent and intense. Still, scientists are uncertain about how drought will affect the natural world, particularly the bacteria, fungi, and other microbes that live in soils. These tiny life forms are crucial because they control the Earth’s flows of carbon and essential nutrients. Researchers at the University of California, Irvine, and Lawrence Berkeley National Laboratory teamed up to study how the microbiome, or collection of bacteria and fungi in the soil, deals with drought. Since 2007, the researchers have used shelters with retractable roofs to prevent nearly half of normal rainfall from reaching grass and shrub ecosystems, and their soil microbiomes, in Southern California. The study team discovered that microbes have some clever tricks up their sleeve for surviving drought. When growing on dead grass as a food source, microbes ramp up production of specialized chemicals called osmolytes that keep their cells from drying out. But microbes growing on dead shrubs face another problem. Unlike grass, shrub leaves are a lousy food source. To digest and consume shrub leaves, microbes have to exude more enzymes, which act like biochemical chef knives that chop complex leaf molecules into bite-sized pieces. Carbon is the coin of the microbial realm, earned via enzyme action or slurping up dead plant juices. Microbes growing under normal conditions on tasty dead grass have it easy: they can spend most of their carbon currency on growth. With drought, life gets harder as microbes need to pay up for osmolytes and ramp down their growth. It gets worse with shrub leaves because microbes have to multi-task among growth, enzyme secretion, and osmolyte production. When drought hits, microbes on shrub leaves forgo the osmolytes, probably because losing the carbon revenue from enzyme investment would be a deal-breaker for survival. The next question tackled by the researchers asked how the genes controlling microbial lifestyles sort out across the tree of life. Most microbiologists thought these lifestyles would correspond to rather large branches on the tree. But the study team found that in fact, very closely related microbes differ in important and interesting ways. For example, bacteria that have nearly identical housekeeping genes respond distinctively to warming, rainfall, and plant chemistry. As a result, soil microbiomes are much richer in diversity than originally thought. And studying microbial diversity in much greater detail could open up many more possibilities for how microbial life deals with changing environmental conditions. Trying to understand microbial life without the fine details of genetic diversity is like trying to stream Netflix on a dial-up connection. By looking across the landscape, the researchers revealed that microbial diversity is absolutely critical for maintaining the planet’s flows of carbon and nutrients. The study team designed a new technology—microbial cages—for transplanting intact microbiomes. With the cages, the researchers could move microbiomes to novel environments and compare their ability to cycle carbon and nutrients. In some cases, performance tailed off when microbiomes found themselves in a new environment, but in other cases, performance rivaled or even exceeded that of the resident microbiome. And even the low performers eventually caught up to the native microbiomes if given sufficient time. These findings mean that microbiomes—at least in Southern California—may be resilient to climate shifts due to a high diversity of microbial lifestyles. Coping with heat and drought may literally be in their DNA. The last piece of the research puzzle, and a “Holy Grail” for microbial ecologists, is to forecast the behavior of diverse microbiomes. To meet this challenge, the study team developed new theory and computer models. For the first time, these models account for hundreds of different microbes and how their intricate lifestyles cope with drought. The models are starting to connect the tiniest microbes with the global cycles that sustain Earth’s farms, fields, and forests. With that connection, it will be easier for society to plan for a world with more droughts and other climate disruptions.

54 ENVIRONMENTAL SCIENCES↗

Application of ecological, geological and oceanographic ERTS-1 imagery to Delaware's coastal resources planning

The author has identified the following significant events. Coastal vegetation species appearing in the ERTS-1 image taken of the Southern Coast of Delaware, during orbit 333 on August 16, 1972, have been correlated with ground truth vegetation maps, and imagery obtained from high altitude RB-57 and U-2 overflights. The vegetation maps of the entire Delaware Coast were prepared using data collected on foot, in small boats, and from low altitude aircraft. Multispectral analysis of high altitude RB-57 and U-2 photographs indicated that five vegetation species could be clearly discriminated from 60,000 feet altitude, including: (1) salt marsh cord grass (Spartina alterniflora); (2) salt marsh hay and spike grass (Spartina patens and Distichlis spicata); (3) reed grass (Phragmites communis); (4) high tide bush and sea myrtle (Iva species and Baccharus halimifolia); and (5) a group of fresh water species found in impounded areas built to attract water fowl. Major Spartina alterniflora and Spartina patens communities within the tidal marshes can be identified in the ERTS-1 imagery. Phragmites, and other species however, occur in smaller, more dispersed groupings and are difficult to discriminate within the resolution capability of the ERTS-1 scanner. Similarly, major impounded areas, built to attract water fowl can be detected; however, mosquito drainage ditches, covering many of Delaware's marshes, are too narrow and not long enough to be resolved by ERTS-1 sensors. High-marsh and dune communities dominated by high tide bush (Iva frutescens) and sea myrtle (Baccharus halimifolia) can be distinguished from adjacent maritime forest and beach grass communities.

Klemas, V.↗

Reflectance of litter accumulation levels at five wavelengths within the 0.5- to 2.5 micron waveband

Reflectance was measured for 1-m2 range grass plots with two canopy treatments (standing and clipped) and four levels of litter accumulation and for grain sorghum with two canopy treatments. Reflectance was significantly higher at the 0.65- to 1.65-, and 2.20-micrometer. Wavelengths for both grass and grain sorghum canopies when the canopies were clipped and the resulting litter was removed. The natural accumulation of litter under the grass canopy did not significantly affect reflectance. The 1.65- and 2.20-micrometer wavelength reflectances of the live grass and the intact litter were 21.8% and 16.2%, respectively, and those of grain sorghum were 21.8% and 16.5%, respectively.

Gerbermann, A. H.↗

Characteristics of AVIRIS Band Measurements in Desert Agroecosystems in the Area of Blythe, California: Studies of Cotton Spectra - 1

Airborne Visible/Infrared Imaging Spectrometer (AVIRIS) data from Blythe, California, were acquired in June 1997 to study agricultural spectra from different crops and to identify crops in other areas with similar environmental factors and similar spectral properties. The main objectives of this study are: (1) to compare the spectral and radiometric characteristics of AVIRIS data from agriculture crops with ground spectra measured by a FieldSpec ASD spectrometer; (2) to explore the use of AVIRIS spectral images for identifying agricultural crops; (3) to study the spectral expression of environmental factors on selected crops; and (4) to build a spectral library for the crops that were studied. A long-term goal is to extend the spectral library for different vegetation or crops in different stages of growth. To support our study, on July 18 and 19, 2000, we collected spectra using the FieldSpec spectrometer from selected fields with different crops in the Blythe area of California (longitude 114 deg 33.28 W and latitude 33 deg 25.42 N to longitude 1140 44.53 W and latitude 33 deg 39.77 N). These crops were cotton in different stages of growth, varieties of grass pure or mixed, Sudan grass, Bermuda grass, Teff grass, and alfalfa. Some of the fields were treated with different types of irrigation (i.e., wet to dry conditions). Additional parameters were studied such as the soil water content (WC), pH, and organic matter (OM). The results of this study showed that for crops known to be similar, there is a significant correlation between the spectra that were collected by AVIRIS in 1997 and spectra measured by the FieldSpec (registered) spectrometer in 2000. This correlation allowed development of a spectral library to be used in ENVI-IDL analysis software. This library was used successfully to identify different crops. Furthermore, using IDL algorithms of Spectral Angle Mapper classification (SAM), spectral feature fitting (SFF) and spectral binary encoding (SPE) showed that there is excellent agreement between the predicted and the actual crop type (i.e., the correlation is between 85-90% match). Further use of the AVIRIS images can be of a value to crop identification or crop yield for commercial use.

Hanna, Safwat H. Shakir↗

Comparative productivity of six bioenergy cropping systems on marginal lands in the Great Lakes Region, United States

Abstract Growing lignocellulosic crops on marginal lands is a promising solution for sustainable biofuel production. We evaluated the productivity of bioenergy cropping systems (switchgrass [ Panicum virgatum L., var. Cave‐In‐Rock], miscanthus [ Miscanthus × giganteus , ‘Illinois clone’], hybrid poplar [ Populus nigra × P. maximowiczii A. Henry ‘NM6’], native grasses [five species], early successional vegetation, and restored prairie vs. historical vegetation [as reference control]) with and without nitrogen fertilization on low‐fertility former cropland at five sites in the Great Lakes Region, United States. We reported biomass yields for the first 7 years after establishment. Switchgrass was most consistently productive across all sites but miscanthus was more productive at three of the five sites. When averaged across sites, years, and nitrogen (N) treatments, biomass yields followed the order miscanthus > switchgrass > hybrid poplar ≈ native grasses > restored prairie > early successional vegetation ≈ historical vegetation, but varied substantially by crop and site, with a significant crop by site interaction. Yields of miscanthus and switchgrass peaked after four–five growing seasons and declined thereafter, while yields of both native grasses and restored prairie increased throughout 6 years with no sign of follow‐on decline, suggesting that polycultures may outperform monocultures over the long term. Yields of early successional vegetation—similar in composition to historical vegetation at each site—did not improve with time. Nitrogen fertilization increased the yields of all cropping systems at all sites. Our results demonstrate the viability of low‐productivity former cropland for long‐term bioenergy production and suggest there is no single crop best suited for all low fertility soils.

Jayawardena, Dileepa M.↗

Whole plant traits of coastal dune vegetation and implications for interactions with dune dynamics

Abstract Coastal dunes are important protective features against sea level rise and coastal storms. Interactions between dune plant aboveground structures and sediment trapping that allow for dune building and maintenance are well established. More recently, studies documenting belowground biomass for promoting erosion resistance in dominant dune species have been conducted, yet a knowledge gap remains regarding species‐specific characterization of whole plants, specifically with respect to roots, rhizomes, and belowground stems. Our objective was to quantify above‐ and belowground traits of four dominant dune grasses to document the potential for species‐specific effects on dune growth, maintenance, and erosion resistance. We examined above‐ and belowground traits among four prominent dune grasses of the Atlantic and Gulf Coasts of North America: Ammophila breviligulata , Panicum amarum , Spartina patens , and Uniola paniculata . Whole plant samples of each species were collected from the foredune at the US Army Engineer Research and Development Center's Field Research Facility in Duck, North Carolina, USA, and quantified for several above‐ and belowground traits (e.g., stem height, rhizome number and length, root surface area by diameter class, root tensile strength, and mycorrhizal percent infection). Belowground factors known to impact important dune processes, such as rhizome length, mycorrhizal percent infection, and root traits, differed substantially among species. When visualized in multivariate space, all species significantly differed in suites of above‐ and belowground traits. When considering belowground only, Ammophila and Spartina were similar, despite differences in biomass allocation. Species separated along axes related to mycorrhizal association, biomass allocation, and root construction. The four co‐occurring dune grass species were dissimilar in suites of plant traits. Belowground trait differences were driven by those describing root construction, biomass allocation, and mycorrhizal infection. Dissimilarity in above‐ and belowground suites of traits may demonstrate different approaches for surviving the dune environment. Incorporating belowground traits into modeling will enhance predictions of dune response to climate change through interactions between vegetation and dune dynamics that facilitate coastal resistance and resilience.

54 ENVIRONMENTAL SCIENCES↗

Root water uptake of biofuel crops revealed by coupled electrical resistivity and soil water content measurements

Abstract Biofuel crops, including annuals such as maize ( Zea mays L.), soybean [ Glycine max (L.) Merr.], and canola ( Brassica napus L.), as well as high‐biomass perennial grasses such as miscanthus ( Miscanthus × giganteus J.M. Greef & Deuter ex Hodkinson & Renvoiz), are candidates for sustainable alternative energy sources. However, large‐scale conversion of croplands to perennial biofuel crops could have substantial impacts on regional water, nutrient, and C cycles due to the longer growing seasons and differences in rooting systems compared with most annual crops. However, due to the limited tools available to nondestructively study the spatiotemporal patterns of root water uptake in situ at field scales, these differences in crop water use are not well known. Geophysical imaging tools such as electrical resistivity (ER) reveal changes in water content in the soil profile. In this study, we demonstrate the use of a novel coupled hydrogeophysical approach with both time domain reflectometry soil water content and ER measurements to compare root water uptake and soil properties of an annual crop rotation with the perennial grass miscanthus, across three growing seasons (2009–2011) in southwest Michigan, USA. We estimated maximum root depths to be between 1.2 and 2.2 m, with the vertical distribution of roots being notably deeper in 2009 relative to 2010 and 2011, likely due to the drought conditions during that first year. Modeled cumulative ET of both crops was underestimated (2–34%) relative to estimates obtained from soil water drawdown in prior studies but was found to be greater in the perennial grass than the annual crops, despite shallower modeled rooting depths in 2010 and 2011.

Kuhl, Alexandria S.↗

Coupling Remote Sensing With a Process Model for the Simulation of Rangeland Carbon Dynamics

Rangelands provide significant environmental benefits through many ecosystem services, which may include soil organic carbon (SOC) sequestration. However, quantifying SOC stocks and monitoring carbon (C) fluxes in rangelands are challenging due to the considerable spatial and temporal variability tied to rangeland C dynamics as well as limited data availability. We developed the Rangeland Carbon Tracking and Management (RCTM) system to track long-term changes in SOC and ecosystem C fluxes by leveraging remote sensing inputs and environmental variable data sets with algorithms representing terrestrial C-cycle processes. Bayesian calibration was conducted using quality-controlled C flux data sets obtained from 61 Ameriflux and NEON flux tower sites from Western and Midwestern US rangelands to parameterize the model according to dominant vegetation classes (perennial and/or annual grass, grass-shrub mixture, and grass-tree mixture). The resulting RCTM system produced higher model accuracy for estimating annual cumulative gross primary productivity (GPP) (R 2 > 0.6, RMSE <390 g C m -2 ) relative to net ecosystem exchange of CO 2 (NEE) (R 2 > 0.4, RMSE <180 g C m -2 ). Model performance in estimating rangeland C fluxes varied by season and vegetation type. The RCTM captured the spatial variability of SOC stocks with R 2 = 0.6 when validated against SOC measurements across 13 NEON sites. Model simulations indicated slightly enhanced SOC stocks for the flux tower sites during the past decade, which is mainly driven by an increase in precipitation. Future efforts to refine the RCTM system will benefit from long-term network-based monitoring of vegetation biomass, C fluxes, and SOC stocks.

54 ENVIRONMENTAL SCIENCES↗

Genome biology of the paleotetraploid perennial biomass crop Miscanthus

Abstract Miscanthus is a perennial wild grass that is of global importance for paper production, roofing, horticultural plantings, and an emerging highly productive temperate biomass crop. We report a chromosome-scale assembly of the paleotetraploid M. sinensis genome, providing a resource for Miscanthus that links its chromosomes to the related diploid Sorghum and complex polyploid sugarcanes. The asymmetric distribution of transposons across the two homoeologous subgenomes proves Miscanthus paleo-allotetraploidy and identifies several balanced reciprocal homoeologous exchanges. Analysis of M. sinensis and M. sacchariflorus populations demonstrates extensive interspecific admixture and hybridization, and documents the origin of the highly productive triploid bioenergy crop M. × giganteus . Transcriptional profiling of leaves, stem, and rhizomes over growing seasons provides insight into rhizome development and nutrient recycling, processes critical for sustainable biomass accumulation in a perennial temperate grass. The Miscanthus genome expands the power of comparative genomics to understand traits of importance to Andropogoneae grasses.

59 BASIC BIOLOGICAL SCIENCES↗

INDETERMINATE1 -mediated expression of FT family genes is required for proper timing of flowering in Brachypodium distachyon

The transition to flowering is a major developmental switch in plants. In many temperate grasses, perception of indicators of seasonal change, such as changing day-length and temperature, leads to expression of FLOWERING LOCUS T1 ( FT1 ) and FT-Like ( FTL ) genes that are essential for promoting the transition to flowering. However, little is known about the upstream regulators of FT1 and FTL genes in temperate grasses. Here, we characterize the monocot-specific gene INDETERMINATE1 ( BdID1 ) in Brachypodium distachyon and demonstrate that BdID1 is a regulator of FT family genes. Mutations in ID1 impact the ability of the short-day (SD) vernalization, cold vernalization, and long-day (LD) photoperiod pathways to induce certain FTL genes. BdID1 is required for upregulation of FTL9 ( FT-LIKE9 ) expression by the SD vernalization pathway, and overexpression of FTL9 in an id1 background can partially restore the delayed flowering phenotype of id1 . We show that BdID1 binds in vitro to the promoter region of FTL genes suggesting that ID1 directly activates FTL expression. Transcriptome analysis shows that BdID1 is required for FT1 , FT2 , FTL12 , and FTL13 expression under inductive LD photoperiods, indicating that BdID1 is a regulator of the FT gene family. Moreover, overexpression of FT1 in the id1 background results in rapid flowering similar to overexpressing FT1 in the wild type, demonstrating that BdID1 is upstream of FT family genes. Interestingly, ID1 negatively regulates a previously uncharacterized FTL gene, FTL4, and we show that FTL4 is a repressor of flowering. Thus, BdID1 is critical for proper timing of flowering in temperate grasses.

59 BASIC BIOLOGICAL SCIENCES↗

Biochemical Characterization of Rice Xylan Biosynthetic Enzymes in Determining Xylan Chain Elongation and Substitutions

Abstract Grass xylan consists of a linear chain of β-1,4-linked xylosyl residues that often form domains substituted only with either arabinofuranose (Araf) or glucuronic acid (GlcA)/methylglucuronic acid (MeGlcA) residues, and it lacks the unique reducing end tetrasaccharide sequence found in dicot xylan. The mechanism of how grass xylan backbone elongation is initiated and how its distinctive substitution pattern is determined remains elusive. Here, we performed biochemical characterization of rice xylan biosynthetic enzymes, including xylan synthases, glucuronyltransferases and methyltransferases. Activity assays of rice xylan synthases demonstrated that they required short xylooligomers as acceptors for their activities. While rice xylan glucuronyltransferases effectively glucuronidated unsubstituted xylohexaose acceptors, they transferred little GlcA residues onto (Araf)-substituted xylohexaoses and rice xylan 3-O-arabinosyltransferase could not arabinosylate GlcA-substituted xylohexaoses, indicating that their intrinsic biochemical properties may contribute to the distinctive substitution patterns of rice xylan. In addition, we found that rice xylan methyltransferase exhibited a low substrate binding affinity, which may explain the partial GlcA methylation in rice xylan. Furthermore, immunolocalization of xylan in xylem cells of both rice and Arabidopsis showed that it was deposited together with cellulose in secondary walls without forming xylan-rich nanodomains. Together, our findings provide new insights into the biochemical mechanisms underlying xylan backbone elongation and substitutions in grass species.

Cell Biology↗

The SvFUL2 transcription factor is required for inflorescence determinacy and timely flowering in Setaria viridis

Inflorescence architecture in cereal crops directly impacts yield potential through regulation of seed number and harvesting ability. Extensive architectural diversity found in inflorescences of grass species is due to spatial and temporal activity and determinacy of meristems, which control the number and arrangement of branches and flowers, and underlie plasticity. Timing of the floral transition is also intimately associated with inflorescence development and architecture, yet little is known about the intersecting pathways and how they are rewired during development. Here, we show that a single mutation in a gene encoding an AP1/FUL-like MADS-box transcription factor significantly delays flowering time and disrupts multiple levels of meristem determinacy in panicles of the C 4 model panicoid grass, Setaria viridis. Previous reports of AP1/FUL-like genes in cereals have revealed extensive functional redundancy, and in panicoid grasses, no associated inflorescence phenotypes have been described. In S. viridis, perturbation of SvFul2, both through chemical mutagenesis and gene editing, converted a normally determinate inflorescence habit to an indeterminate one, and also repressed determinacy in axillary branch and floral meristems. Our analysis of gene networks connected to disruption of SvFul2 identified regulatory hubs at the intersection of floral transition and inflorescence determinacy, providing insights into the optimization of cereal crop architecture.

59 BASIC BIOLOGICAL SCIENCES↗

Transcriptome and DNA methylome divergence of inflorescence development between 2 ecotypes in Panicum hallii

The morphological diversity of the inflorescence determines flower and seed production, which is critical for plant adaptation. Hall's panicgrass (Panicum hallii, P. hallii) is a wild perennial grass that has been developed as a model to study perennial grass biology and adaptive evolution. Highly divergent inflorescences have evolved between the 2 major ecotypes in P. hallii, the upland ecotype (P. hallii var hallii, HAL2 genotype) with compact inflorescence and large seed and the lowland ecotype (P. hallii var filipes, FIL2 genotype) with an open inflorescence and small seed. Here we conducted a comparative analysis of the transcriptome and DNA methylome, an epigenetic mark that influences gene expression regulation, across different stages of inflorescence development using genomic references for each ecotype. Global transcriptome analysis of differentially expressed genes (DEGs) and co-expression modules underlying the inflorescence divergence revealed the potential role of cytokinin signaling in heterochronic changes. Comparing DNA methylome profiles revealed a remarkable level of differential DNA methylation associated with the evolution of P. hallii inflorescence. We found that a large proportion of differentially methylated regions (DMRs) were located in the flanking regulatory regions of genes. Intriguingly, we observed a substantial bias of CHH hypermethylation in the promoters of FIL2 genes. The integration of DEGs, DMRs, and $K_a$/$K_s$ ratio results characterized the evolutionary features of DMR-associated DEGs that contribute to the divergence of the P. hallii inflorescence. This study provides insights into the transcriptome and epigenetic landscape of inflorescence divergence in P. hallii and a genomic resource for perennial grass biology.

59 BASIC BIOLOGICAL SCIENCES↗

Disruption of p -coumaroyl-CoA:monolignol transferases in rice drastically alters lignin composition

Grasses are abundant feedstocks that can supply lignocellulosic biomass for production of cell-wall-derived chemicals. In grass cell walls, lignin is acylated with p-coumarate. These p-coumarate decorations arise from the incorporation of monolignol p-coumarate conjugates during lignification. A previous biochemical study identified a rice (Oryza sativa) BAHD acyltransferase (AT) with p-coumaroyl-CoA:monolignol transferase (PMT) activity in vitro. In this study, we determined that that enzyme, which we name OsPMT1 (also known as OsAT4), and the closely related OsPMT2 (OsAT3) harbor similar catalytic activity toward monolignols. We generated rice mutants deficient in either or both OsPMT1 and OsPMT2 by CRISPR/Cas9-mediated mutagenesis and subjected the mutants’ cell walls to analysis using chemical and nuclear magnetic resonance methods. Our results demonstrated that OsPMT1 and OsPMT2 both function in lignin p-coumaroylation in the major vegetative tissues of rice. Notably, lignin-bound p-coumarate units were undetectable in the ospmt1 ospmt2-2 double-knockout mutant. Further, in-depth structural analysis of purified lignins from the ospmt1 ospmt2-2 mutant compared with control lignins from wild-type rice revealed stark changes in polymer structures, including alterations in syringyl/guaiacyl aromatic unit ratios and inter-monomeric linkage patterns, and increased molecular weights. Furthermore, our results provide insights into lignin polymerization in grasses that will be useful for the optimization of bioengineering approaches for the effective use of biomass in biorefineries.

59 BASIC BIOLOGICAL SCIENCES↗