CELLULOSE SYNTHASE-LIKE C proteins modulate cell wall establishment during ethylene-mediated root growth inhibition in rice.

The cell wall shapes plant cell morphogenesis and affects the plasticity of organ growth. However, the way in which cell wall establishment is regulated by ethylene remains largely elusive. Here, by analyzing cell wall patterns, cell wall composition and gene expression in rice (Oryza sativa, L.) roots, we found that ethylene induces cell wall thickening and the expression of cell wall synthesis-related genes, including CELLULOSE SYNTHASE-LIKE C1, 2, 7, 9, 10 (OsCSLC1, 2, 7, 9, 10) and CELLULOSE SYNTHASE A3, 4, 7, 9 (OsCESA3, 4, 7, 9). Overexpression and mutant analyses revealed that OsCSLC2 and its homologs function in ethylene-mediated induction of xyloglucan biosynthesis mainly in the cell wall of root epidermal cells. Moreover, OsCESA-catalyzed cellulose deposition in the cell wall was enhanced by ethylene. OsCSLC-mediated xyloglucan biosynthesis likely plays an important role in restricting cell wall extension and cell elongation during the ethylene response in rice roots. Genetically, OsCSLC2 acts downstream of ETHYLENE-INSENSITIVE3-LIKE1 (OsEIL1)-mediated ethylene signaling, and OsCSLC1, 2, 7, 9 are directly activated by OsEIL1. Furthermore, the auxin signaling pathway is synergistically involved in these regulatory processes. These findings link plant hormone signaling with cell wall establishment, broadening our understanding of root growth plasticity in rice and other crops.

Natural zeolite enhances anaerobic digestion of waste activated sludge: Insights into the performance and the role of biofilm.

Anaerobic digestion is widely employed for the treatment of waste activated sludge (WAS) due to its advantages like simultaneous energy recovery and sludge stabilization, promoting carbon-neutral operation of wastewater treatment plants. Natural zeolite, a low-cost and eco-friendly additive, has the potential to improve methane production from anaerobic digestion. This study investigated the effects of natural zeolite on anaerobic digestion when the substrate was WAS. It was found that methane production potential in response to natural zeolite was dosage-dependent. The optimal dosage was 0.1 g zeolite/g volatile suspended solids (VSS), with a methane yield of 181.89 ± 6.75 mL/g VSS, which increased by 20.1% compared to that of the control. Although the methane yields with other dosages of natural zeolite were higher than that of control, they were lesser than that with 0.1 g zeolite/g VSS. Natural zeolite affected transfer and conversion of proteins much more than polysaccharides in liquid phase and extracellular polymeric substances. In anaerobic digestion, natural zeolite had with little effects on WAS solubilization, while it improved hydrolysis, acidification, and methanogenesis. The dosages of natural zeolite did have significant effects on bacterial communities in biofilm rather than suspension, while the archaeal communities in biofilm and suspension were all greatly related to natural zeolite dosages. The developed biofilms promoted richness and functionality of microbial communities. The syntrophic metabolism relationships between methanogens and bacteria were improved, which was proved by selective enrichment of Methanosarcina, Syntrophomonas, and Petrimonas. The findings of this work provided some new solutions for promoting methane production from WAS, and the roles of natural zeolite in anaerobic digestion.

Insights into response mechanism of anaerobic digestion of waste activated sludge to particle sizes of zeolite.

Anaerobic digestion has been proved as one promising strategy to simultaneously achieve resource recovery and environmental pollution control for biosolid treatment, and adding exogenous materials is a potential alternative to promote the above process. This study investigated response mechanisms of anaerobic digestion of waste activated sludge (WAS) to particle sizes of zeolite. Results showed that the methane production reached 186.75 ± 7.62 mL/g volatile suspended solids (VSS) with zeolite of the particle size of 0.2-0.5 mm and the additive dosage of 0.1 g/g VSS, which increased by 22.08% compared to that in control. Mechanism study revealed that zeolite could improve hydrolysis, acidification, and methanogenesis stages. Rapid consumption rates of soluble polysaccharides and proteins were observed, correspondingly, the accumulations of short-chain fatty acids (SCFAs) were enhanced, and the compositions of SCFAs were optimized. Moreover, the activities of F420 increased by 28% with zeolite, and the syntrophic metabolism between bacteria and methanogens were promoted.

Plant-on-chip: Core morphogenesis processes in the tiny plant Wolffia australiana.

A plant can be thought of as a colony comprising numerous growth buds, each developing to its own rhythm. Such lack of synchrony impedes efforts to describe core principles of plant morphogenesis, dissect the underlying mechanisms, and identify regulators. Here, we use the minimalist known angiosperm to overcome this challenge and provide a model system for plant morphogenesis. We present a detailed morphological description of the monocot Wolffia australiana, as well as high-quality genome information. Further, we developed the plant-on-chip culture system and demonstrate the application of advanced technologies such as single-nucleus RNA-sequencing, protein structure prediction, and gene editing. We provide proof-of-concept examples that illustrate how W. australiana can decipher the core regulatory mechanisms of plant morphogenesis.

Galactosylation of rhamnogalacturonan-II for cell wall pectin biosynthesis is critical for root apoplastic iron reallocation in Arabidopsis.

Apoplastic iron (Fe) in roots represents an essential Fe storage pool. Reallocation of apoplastic Fe is of great importance to plants experiencing Fe deprivation, but how this reallocation process is regulated remains elusive, likely because of the highly complex cell wall structure and the limited knowledge about cell wall biosynthesis and modulation. Here, we present genetic and biochemical evidence to demonstrate that the Cdi-mediated galactosylation of rhamnogalacturonan-II (RG-II) is required for apoplastic Fe reallocation. Cdi is expressed in roots and up-regulated in response to Fe deficiency. It encodes a putative glycosyltransferase localized to the Golgi apparatus. Biochemical and mass spectrometry assays showed that Cdi catalyzes the transfer of GDP-L-galactose to the terminus of side chain A on RG-II. Disruption of Cdi essentially decreased RG-II dimerization and hence disrupted cell wall formation, as well as the reallocation of apoplastic Fe from roots to shoots. Further transcriptomic, Fourier transform infrared spectroscopy, and Fe desorption kinetic analyses coincidently suggested that Cdi mediates apoplastic Fe reallocation through extensive modulation of cell wall components and consequently the Fe adsorption capacity of the cell wall. Our study provides direct evidence demonstrating a link between cell wall biosynthesis and apoplastic Fe reallocation, thus indicating that the structure of the cell wall is important for efficient usage of the cell wall Fe pool.

Research on the Effect of Molten Salt Ultrasonic Composite Cleaning for Paint Removal.

Aiming at paint removal on hydraulic cylinder, the effect of molten salt ultrasonic composite cleaning was studied. First, the mechanism of molten salt cleaning and ultrasonic cleaning was reviewed. To further describe the composite cleaning mechanism, the components and internal structure of paint were analyzed by scanning electron microscopy and Fourier transform infrared. Results showed that the paint had a significant layered structure. The total thickness was about 100 µm, and the main components were organic matters, including ester groups, epoxy groups, and aromatic compounds. Then, combining with thermal environment, cleaning medium's property, and ultrasound, the composite cleaning mechanism was described in terms of three aspects: thermal effect, chemical reaction, and ultrasonic effect. Besides, the reason why this composite cleaning had good effect on paint removal, compared to paint heated in air, was explained through dynamic analysis, which was the reduction of reaction activation energy from 114.4 kJ/mol of paint alone to 74.1 kJ/mol.

Xyloglucan Fucosylation Modulates Arabidopsis Cell Wall Hemicellulose Aluminium binding Capacity.

Although xyloglucan (XyG) is reported to bind Aluminium (Al), the influence of XyG fucosylation on the cell wall Al binding capacity and plant Al stress responses is unclear. We show that Arabidopsis T-DNA insertion mutants with reduced AXY3 (XYLOSIDASE1) function and consequent reduced levels of fucosylated XyG are more sensitive to Al than wild-type Col-0 (WT). In contrast, T-DNA insertion mutants with reduced AXY8 (FUC95A) function and consequent increased levels of fucosylated XyG are more Al resistant. AXY3 transcript levels are strongly down regulated in response to 30 min Al treatment, whilst AXY8 transcript levels also repressed until 6 h following treatment onset. Mutants lacking AXY3 or AXY8 function exhibit opposing effects on Al contents of root cell wall and cell wall hemicellulose components. However, there was no difference in the amount of Al retained in the pectin components between mutants and WT. Finally, whilst the total sugar content of the hemicellulose fraction did not change, the altered hemicellulose Al content of the mutants is shown to be a likely consequence of their different XyG fucosylation levels. We conclude that variation in XyG fucosylation levels influences the Al sensitivity of Arabidopsis by affecting the Al-binding capacity of hemicellulose.

TRICHOME BIREFRINGENCE-LIKE27 affects aluminum sensitivity by modulating the O-acetylation of xyloglucan and aluminum-binding capacity in Arabidopsis.

Xyloglucan (XyG) has been reported to contribute to the aluminum (Al)-binding capacity of the cell wall in Arabidopsis (Arabidopsis thaliana). However, the influence of O-acetylation of XyG, accomplished by the putative O-acetyltransferase TRICHOME BIREFRINGENCE-LIKE27 (TBL27 [AXY4]), on its Al-binding capacity is not known. In this study, we found that the two corresponding TBL27 mutants, axy4-1 and axy4-3, were more Al sensitive than wild-type Columbia-0 plants. TBL27 was expressed in roots as well as in leaves, stems, flowers, and siliques. Upon Al treatment, even within 30 min, TBL27 transcript accumulation was strongly down-regulated. The mutants axy4-1 and axy4-3 accumulated significantly more Al in the root and wall, which could not be correlated with pectin content or pectin methylesterase activity, as no difference in the mutants was observed compared with the wild type when exposed to Al stress. The increased Al accumulation in the wall of the mutants was found to be in the hemicellulose fraction. While the total sugar content of the hemicellulose fraction did not change, the O-acetylation level of XyG was reduced by Al treatment. Taken together, we conclude that modulation of the O-acetylation level of XyG influences the Al sensitivity in Arabidopsis by affecting the Al-binding capacity in the hemicellulose.

Disruption of secondary wall cellulose biosynthesis alters cadmium translocation and tolerance in rice plants.

Tricheary elements (TEs), wrapped by secondary cell wall, play essential roles in water, mineral, and nutrient transduction. Cadmium (Cd) is a toxic heavy metal that is absorbed by roots and transported to shoot, leaves, and grains through vascular systems in plants. As rice is a major source of Cd intake, many efforts have been made to establish 'low-Cd rice'. However, no links have been found between cellulose biosynthesis and cadmium accumulation. We report here a rice brittle culm13 mutant, resulting from a novel missense mutation (E101K) [corrected] in the N-terminus of cellulose synthase subunit 9 (CESA9). Except for the abnormal mechanical strength, the mutant plants are morphologically indistinguishable from the wild-type plants. Transmission electron microscopy (TEM) and chemical analyses showed a slight reduction in secondary wall thickness and 22% decrease in cellulose content in bc13 plants. Moreover, this mutation unexpectedly confers the mutant plants Cd tolerance due to less Cd accumulation in leaves. Expression analysis of the genes required for Cd uptake and transport revealed complicated alterations after applying Cd to wild-type and bc13. The mutants were further found to have altered vascular structure. More importantly, Cd concentration in the xylem saps from the bc13 plants was significantly lower than that from the wild-type. Combining the analyses of CESA9 gene expression and Cd content retention in the cell-wall residues, we conclude that CESA9(E101K) [corrected] mutation alters cell-wall properties in the conducting tissues, which consequently affects Cd translocation efficiency that largely contributes to the low Cd accumulation in the mutant plants.

The transcription factor AtDOF4.2 regulates shoot branching and seed coat formation in Arabidopsis.

Plant-specific DOF (DNA-binding with one finger)-type transcription factors regulate various biological processes. In the present study we characterized a silique-abundant gene AtDOF (Arabidopsis thaliana DOF) 4.2 for its functions in Arabidopsis. AtDOF4.2 is localized in the nuclear region and has transcriptional activation activity in both yeast and plant protoplast assays. The T-M-D motif in AtDOF4.2 is essential for its activation. AtDOF4.2-overexpressing plants exhibit an increased branching phenotype and mutation of the T-M-D motif in AtDOF4.2 significantly reduces branching in transgenic plants. AtDOF4.2 may achieve this function through the up-regulation of three branching-related genes, AtSTM (A. thaliana SHOOT MERISTEMLESS), AtTFL1 (A. thaliana TERMINAL FLOWER1) and AtCYP83B1 (A. thaliana CYTOCHROME P450 83B1). The seeds of an AtDOF4.2-overexpressing plant show a collapse-like morphology in the epidermal cells of the seed coat. The mucilage contents and the concentration and composition of mucilage monosaccharides are significantly changed in the seed coat of transgenic plants. AtDOF4.2 may exert its effects on the seed epidermis through the direct binding and activation of the cell wall loosening-related gene AtEXPA9 (A. thaliana EXPANSIN-A9). The dof4.2 mutant did not exhibit changes in branching or its seed coat; however, the silique length and seed yield were increased. AtDOF4.4, which is a close homologue of AtDOF4.2, also promotes shoot branching and affects silique size and seed yield. Manipulation of these genes should have a practical use in the improvement of agronomic traits in important crops.

XTH31, encoding an in vitro XEH/XET-active enzyme, regulates aluminum sensitivity by modulating in vivo XET action, cell wall xyloglucan content, and aluminum binding capacity in Arabidopsis.

Xyloglucan endohydrolase (XEH) and xyloglucan endotransglucosylase (XET) activities, encoded by xyloglucan endotransglucosylase-hydrolase (XTH) genes, are involved in cell wall extension by cutting or cutting and rejoining xyloglucan chains, respectively. However, the physiological significance of this biochemical activity remains incompletely understood. Here, we find that an XTH31 T-DNA insertion mutant, xth31, is more Al resistant than the wild type. XTH31 is bound to the plasma membrane and the encoding gene is expressed in the root elongation zone and in nascent leaves, suggesting a role in cell expansion. XTH31 transcript accumulation is strongly downregulated by Al treatment. XTH31 expression in yeast yields a protein with an in vitro XEH:XET activity ratio of >5000:1. xth31 accumulates significantly less Al in the root apex and cell wall, shows remarkably lower in vivo XET action and extractable XET activity, has a lower xyloglucan content, and exhibits slower elongation. An exogenous supply of xyloglucan significantly ameliorates Al toxicity by reducing Al accumulation in the roots, owing to the formation of an Al-xyloglucan complex in the medium, as verified by an obvious change in chemical shift of (27)Al-NMR. Taken together, the data indicate that XTH31 affects Al sensitivity by modulating cell wall xyloglucan content and Al binding capacity.

Arabidopsis NMD3 is required for nuclear export of 60S ribosomal subunits and affects secondary cell wall thickening.

NMD3 is required for nuclear export of the 60S ribosomal subunit in yeast and vertebrate cells, but no corresponding function of NMD3 has been reported in plants. Here we report that Arabidopsis thaliana NMD3 (AtNMD3) showed a similar function in the nuclear export of the 60S ribosomal subunit. Interference with AtNMD3 function by overexpressing a truncated dominant negative form of the protein lacking the nuclear export signal sequence caused retainment of the 60S ribosomal subunits in the nuclei. More interestingly, the transgenic Arabidopsis with dominant negative interference of AtNMD3 function showed a striking failure of secondary cell wall thickening, consistent with the altered expression of related genes and composition of cell wall components. Observation of a significant decrease of rough endoplasmic reticulum (RER) in the differentiating interfascicular fiber cells of the transgenic plant stems suggested a link between the defective nuclear export of 60S ribosomal subunits and the abnormal formation of the secondary cell wall. These findings not only clarified the evolutionary conservation of NMD3 functions in the nuclear export of 60S ribosomal subunits in yeast, animals and plants, but also revealed a new facet of the regulatory mechanism underlying secondary cell wall thickening in Arabidopsis. This new facet is that the nuclear export of 60S ribosomal subunits and the formation of RER may play regulatory roles in coordinating protein synthesis in cytoplasm and transcription in nuclei.

Identification of quantitative trait loci affecting hemicellulose characteristics based on cell wall composition in a wild and cultivated rice species.

Cell wall hemicellulosic polysaccharides are structurally complex and diverse. Knowledge about the synthesis of cell wall hemicelluloses and their biological roles is limited. Quantitative trait loci (QTL) mapping is a helpful tool for the dissection of complex phenotypes for gene identification. In this study, we exploited the natural variation in cell wall monosaccharide levels between a common wild rice, Yuanj, and an elite indica cultivar, Teqing, and performed QTL mapping with their introgression lines (ILs). Chemical analyses conducted on the culms of Yuanj and Teqing showed that the major alterations are found in glucose and xylose levels, which are correlated with specific hemicellulosic polymers. Glycosidic linkage examination revealed that, in Yuanj, an increase in glucose content results from a higher level of mixed linkage ß-glucan (MLG), whereas a reduction in xylose content reflects a low level of xylan backbone and a varied arabinoxylan (AX) structure. Seventeen QTLs for monosaccharides have been identified through composition analysis of the culm residues of 95 core ILs. Four major QTLs affecting xylose and glucose levels are responsible for 19 and 21% of the phenotypic variance, respectively. This study provides a unique resource for the genetic dissection of rice cell wall formation and remodeling in the vegetative organs.

Aerobic and anaerobic nonmicrobial methane emissions from plant material.

Methane (CH(4)) may be generated via microbial and nonmicrobial mechanisms. Nonmicrobial CH(4) is also ubiquitous in nature, such as in biomass burning, the Earth's crust, plants, and animals. Relative to microbial CH(4), nonmicrobial CH(4) is less understood. Using fresh (living) and dried (dead) leaves and commercial structural compounds (dead) of plants, a series of laboratory experiments have been conducted to investigate CH(4) emissions under aerobic and anaerobic conditions. CH(4) emissions from fresh leaves incubated at ambient temperatures were nonmicrobial and enhanced by anaerobic conditions. CH(4) emissions from dried leaves incubated at rising temperature ruled out a microbial-mediated formation pathway and were plant-species-dependent with three categories of response to oxygen levels: enhanced by aerobic conditions, similar under aerobic and anaerobic conditions, and enhanced by anaerobic conditions. CH(4) emissions in plant structural compounds may help to fully understand nonmicrobial CH(4) formation in plant leaves. Experiments of reactive oxygen species (ROS) generator and scavengers indicate that ROS had a significant role in nonmicrobial CH(4) formation in plant material under aerobic and anaerobic conditions. However, the detailed mechanisms of the ROS were uncertain.

[Cotton fiber quality traits were controlled mainly by maternal plant genotype].

Hai1, a Gossypium barbadense L. variety with super fiber quality, and CCRI36 and Zhong221, two upland cotton cultivars (Gossypium hirsutum L.), were used as recurrent parents to develop two backcross combinations of CCRI36xHai1 and Zhong221xHai1. Fiber quality of inter-crossing bolls and self-crossing bolls were analyzed from different generations of the two combinations. The results showed that there existed significant difference in the average value, pole difference and CV% of fiber quality traits, and no significant correlation in fiber quality traits between inter-crossing bolls (BC2F0) and self-crossing bolls (BC1F1) from male parent plants. There existed no significant difference in the average value, pole difference and CV% of fiber quality traits between inter-crossing bolls (BC2F0) and self-crossing bolls from the recurrent parents when BC1F1 plants were used as male parents and the recurrent parents (CCRI36, Zhong221) as female parents. The results also showed that average value, pole difference and CV% of fiber traits of inter-crossing bolls (BC3F0) were close to those of the female parents (BC2F1). When BC2F1 populations were used as female parents and the recurrent parents (CCRI36, Zhong221) were used as male parents, there were extremely significant positive correlation for fiber length, strength, micronaire value and elongation, except for fiber uniformity between inter-crossing bolls (BC3F0) and self-crossing bolls (BC2F1). So, fiber quality of inter-crossing bolls were close to those of self-crossing bolls of maternal plants and the male parent pollen genotype had no prominent effect to fiber quality traits of inter-crossing bolls. Fiber quality traits were controlled mainly by maternal plant genotype, while the contemporary seed embryonal genotype showed no significant effects for fiber quality.