AtSYP71 Is Important for Plant Cell Wall Biosynthesis and Stress Adaptation.

Background: SYP71, the plant-specific Qc-SNARE protein, is reported to regulate vesicle trafficking. SYP71 is localized on the ER, endosome, plasma membrane and cell plate, suggesting its multiple functions. Lotus SYP71 is essential for symbiotic nitrogen fixation in nodules. AtSYP71, GmSYP71 and OsSYP71 are implicated in plant resistance to pathogenesis. To date, SYP71 regulatory role on plant development remain unclear.Results: AtSYP71-knockout mutant atsyp71-4 was lethal at early development stage. Early development of AtSYP71-knockdown mutant atsyp71-2 was delayed, and stress response was also affected. Confocal images revealed that protein secretion was blocked in atsyp71-2. Transcriptomic analysis indicated that metabolism, response to environmental stimuli pathways and apoplast components were influenced in atsyp71-2. Moreover, the contents of lignin, cellulose and flavonoids as well as cell wall structures were also altered.Conclusion: Our findings suggested that AtSYP71 is essential for plant development. AtSYP71 probably regulates plant development, metabolism and environmental adaptation by affecting cell wall homeostasis via mediating secretion of materials and regulators required for cell wall biosynthesis and dynamics.

Comparative transcriptome and DNA methylation analysis in temperature-sensitive genic male sterile wheat BS366

Background Known as the prerequisite component for the heterosis breeding system, the male sterile line determines the hybrid yield and seed purity. Therefore, a deep understanding of the mechanism and gene network that leads to male sterility is crucial. BS366, a temperature-sensitive genic male sterile (TGMS) line, is male sterile under cold conditions (12 °C with 12 h of daylight) but fertile under normal temperature (20 °C with 12 h of daylight). Results During meiosis, BS366 was defective in forming tetrads and dyads due to the abnormal cell plate. During pollen development, unusual vacuolated pollen that could not accumulate starch grains at the binucleate stage was also observed. Transcriptome analysis revealed that genes involved in the meiotic process, such as sister chromatid segregation and microtubule-based movement, were repressed, while genes involved in DNA and histone methylation were induced in BS366 under cold conditions. MethylRAD was used for reduced DNA methylation sequencing of BS366 spikes under both cold and control conditions. The differentially methylated sites (DMSs) located in the gene region were mainly involved in carbohydrate and fatty acid metabolism, lipid metabolism, and transport. Differentially expressed and methylated genes were mainly involved in cell division. Conclusions These results indicated that the methylation of genes involved in carbon metabolism or fatty acid metabolism might contribute to male sterility in BS366 spikes, providing novel insight into the molecular mechanism of wheat male sterility.

The Glycine-Rich Domain Protein GRDP2 Regulates Ovule Development via the Auxin Pathway in Arabidopsis

The glycine-rich domain proteins (GRDP) have been functionally implicated in the cell wall structure, biotic, and abiotic stress responses. However, little is known about GRDP genes in female gametophyte development of Arabidopsis. This study shows that GRDP2, a GRDP, plays a crucial role in female gametophyte development. In GRDP2 overexpression lines, grdp2-3, the embryo sacs were arrested at FG1 and no nucleus stages. Furthermore, callose staining shows that cell plate formation during megasporogenesis is disturbed in grdp2-3. In contrast, the pollen development is not affected in grdp2-3. The expression patterns of auxin-specific marker lines in female gametophytes showed that the auxin distribution and transport were significantly changed during megagametogenesis in grdp2-3. In addition, compared with the membrane-localized pattern of PIN1, PIN2, and PIN7 in WT, the signals were detected in the cytoplasm in grdp2-3. Together, our data suggest that GRDP2 plays an essential role in auxin-mediated female gametophyte development.

Cell biology of primary cell wall synthesis in plants

Building a complex structure such as the cell wall, with many individual parts that need to be assembled correctly from distinct sources within the cell, is a well-orchestrated process. Additional complexity is required to mediate dynamic responses to environmental and developmental cues. Enzymes, sugars and other cell wall components are constantly and actively transported to and from the plasma membrane during diffuse growth. Cell wall components are transported in vesicles on cytoskeletal tracks composed of microtubules and actin filaments. Many of these components, and additional proteins, vesicles, and lipids are trafficked to and from the cell plate during cytokinesis. In this review, we first discuss how the cytoskeleton is initially organized to add new cell wall material or to build a new cell wall, focusing on similarities during these processes. Next, we discuss how polysaccharides and enzymes that build the cell wall are trafficked to the correct location by motor proteins and through other interactions with the cytoskeleton. Finally, we discuss some of the special features of newly formed cell walls generated during cytokinesis.

The OPAQUE1/DISCORIDA2 myosin XI is required for phragmoplast guidance during asymmetric cell division in maize

Asymmetric divisions produce cells with different fates and are critical for development. Here we show that a maize myosin XI protein, OPAQUE1 (O1), is necessary for asymmetric divisions during maize stomatal development. We analyzed stomatal precursor cells prior to and during asymmetric division to determine why o1 mutants have abnormal division planes. Cell polarization and nuclear positioning occur normally in the o1 mutant, and the future site of division is correctly specified. The defect in o1 occurs during late cytokinesis, when the plant-specific phragmoplast - made of microtubules, actin and other proteins - forms the nascent cell plate. The phragmoplast becomes misguided and does not meet the previously established division site. Initial phragmoplast guidance is correct in o1. However, as phragmoplast expansion continues, phragmoplasts in o1 stomatal precursor cells become misguided and do not meet the cortex at the established division site. To understand how this myosin protein contributes to phragmoplast guidance, we identified O1-interacting proteins. Other myosins, specific actin-binding proteins, and maize kinesins related to the Arabidopsis thaliana division site markers PHRAGMOPLAST ORIENTING KINESINs (POKs) interact with O1. We propose that different myosins are important at multiple steps of phragmoplast expansion, and the O1 actin motor and POK-like microtubule motors work together to ensure correct late-stage phragmoplast guidance.

Comparative Transcriptome and DNA Methylation Analysis in Temperature-Sensitive Genic Male Sterile Wheat BS366

Background: Known as the prerequisite component for the heterosis breeding system, the male sterile line determined the hybrid yield and seed purity. Therefore, a deep understanding of the mechanism and gene network that leads to male sterility is crucial. BS366, a temperature-sensitive genic male sterile (TGMS) line, were male sterile under cold condition (12℃ with 12 h daylight) but fertile under normal temperature (20℃ with 12 h daylight). Results: During the meiosis, BS366 was defective in forming tetrad and dyad due to the abnormal cell plate. During the pollen development, unusual vacuolated pollen that cannot accumulate starch grains at the binucleate stage was also observed. Transcriptome analysis revealed that genes involved in the meiotic process like sister chromatid segregation and microtubule-based movement were repressed, while genes involved in DNA and histone methylation were induced in BS366 under cold condition. MethylRAD was used for a reduced DNA methylation sequencing for BS366 under both cold and control conditions. The differentially methylated sites (DMSs) located in the gene region were mainly involved in carbohydrate and fatty acid metabolism, lipid metabolism, and transport. Genes differentially and methylated were mainly involved in cell division. Conclusions: These results indicated that methylation of genes involved in carbon metabolism or fatty acid metabolism would contribute to the male sterility in BS366, which will provide a novel insight into the molecular mechanism of wheat male sterility.

Dynamic Apical-Basal Enrichment of the F-Actin during Cytokinesis in Arabidopsis Cells Embedded in Their Tissues

During the life cycle of any multicellular organism, cell division contributes to the proliferation of the cell in the tissues as well as the generation of specialized cells, both necessary to form a functional organism. Therefore, the mechanisms of cell division need to be tightly regulated, as malfunctions in their control can lead to tumor formation or developmental defects. This is particularly true in land plants, where cells cannot relocate and therefore cytokinesis is key for morphogenesis. In the green lineage, cell division is executed in radically different manners than animals, with the appearance of new structures (the preprophase band (PPB), cytokinetic the cell plate and phragmoplast), and the disappearance of ancestral mechanisms (cleavage, centrosomes). While F-actin and microtubules closely co-exist to allow the orientation and the progression of the plant cell division, recent studies mainly focused on the involvement of microtubules in this key process. Here, we used our recently developed root tracking system to follow actin dynamics in dividing Arabidopsis meristematic root cells. In this study, we imaged in time and space the fluorescent-tagged F-actin reporter Lifeact together with cell division markers in dividing cells embedded in their tissues. In addition to the F-actin accumulation in the phragmoplasts, we observed and quantified a dynamic apical-basal enrichment of the F-actin during cytokinesis. The role and the possible actors responsible for F-actin dynamics during cytokinesis are discussed.

The effect of geometric imperfections on the mechanical response of isotropic closed-cell plate-lattices

Cubic+octet plate-lattices, whose unit cell comprises plates aligned along simple-cubic and face-centered-cubic planes of crystal structures, have attracted scientists and engineers because of their isotropic, near-optimal mass-specific performance that reaches theoretical upper bounds on stiffness and strength at low density. While their structural efficiency has been recently examined analytically, numerically and experimentally, their sensitivity to geometric imperfections has remained elusive. Here, using finite element simulations, we present sensitivity of the macroscopic mechanical properties of the plate-lattices to two types of geometric imperfections: namely, periodically distributed defects, characterizing plate waviness and displaced intersections, and randomly dispersed defects, representing missing plates, observed in their additively manufactured samples. Our results show that the randomly dispersed imperfections lead to a greater reduction in the Young’s modulus and yield strength than its counterpart across all relative densities under consideration, while their scaling relations with the relative density remains linear, confirming their structural efficiency attributed to stretching-dominated behavior even with the presence of the imperfections. This study sheds light on previously elusive sensitivity of the plate-lattices to the geometric imperfections and provides understanding of their defect-dependent mechanical performance.