Brassinosteroid signals cooperate with katanin-mediated microtubule severing to control stamen filament elongation.

Proper stamen filament elongation is essential for pollination and plant reproduction. Plant hormones are extensively involved in every stage of stamen development; however, the cellular mechanisms by which phytohormone signals couple with microtubule dynamics to control filament elongation remain unclear. Here, we screened a series of Arabidopsis thaliana mutants showing different microtubule defects and revealed that only those unable to sever microtubules, lue1 and ktn80.1234, displayed differential floral organ elongation with less elongated stamen filaments. Prompted by short stamen filaments and severe decrease in KTN1 and KTN80s expression in qui-2 lacking five BZR1-family transcription factors (BFTFs), we investigated the crosstalk between microtubule severing and brassinosteroid (BR) signaling. The BFTFs transcriptionally activate katanin-encoding genes, and the microtubule-severing frequency was severely reduced in qui-2. Taken together, our findings reveal how BRs can regulate cytoskeletal dynamics to coordinate the proper development of reproductive organs.

A novel PGPR strain, Streptomyces lasalocidi JCM 3373T, alleviates salt stress and shapes root architecture in soybean by secreting indole-3-carboxaldehyde.

While soybean (Glycine max L.) provides the most important source of vegetable oil and protein, it is sensitive to salinity, which seriously endangers the yield and quality during soybean production. The application of Plant Growth-Promoting Rhizobacteria (PGPR) to improve salt tolerance for plant is currently gaining increasing attention. Streptomycetes are a major group of PGPR. However, to date, few streptomycetes has been successfully developed and applied to promote salt tolerance in soybean. Here, we discovered a novel PGPR strain, Streptomyces lasalocidi JCM 3373T, from 36 strains of streptomycetes via assays of their capacity to alleviate salt stress in soybean. Microscopic observation showed that S. lasalocidi JCM 3373T does not colonise soybean roots. Chemical analysis confirmed that S. lasalocidi JCM 3373T secretes indole-3-carboxaldehyde (ICA1d). Importantly, IAC1d inoculation alleviates salt stress in soybean and modulates its root architecture by regulating the expression of stress-responsive genes GmVSP, GmPHD2 and GmWRKY54 and root growth-related genes GmPIN1a, GmPIN2a, GmYUCCA5 and GmYUCCA6. Taken together, the novel PGPR strain, S. lasalocidi JCM 3373T, alleviates salt stress and improves root architecture in soybean by secreting ICA1d. Our findings provide novel clues for the development of new microbial inoculant and the improvement of crop productivity under salt stress.

Verticillium dahliae Asp1 regulates the transition from vegetative growth to asexual reproduction by modulating microtubule dynamic organization.

Verticillium dahliae is a devastating pathogenic fungus that causes severe vascular wilts in more than 400 dicotyledonous plants. The conidiation of V. dahliae in plant vascular tissues is the key strategy for its adaptation to the nutrient-poor environment and is required for its pathogenicity. However, it remains unclear about the regulatory mechanism of conidium production of V. dahliae in vascular tissues. Here, we found that VdAsp1, encoding an inositol polyphosphate kinase, is indispensable for the pathogenicity of V. dahliae. Loss of VdAsp1 function does not affect the invasion of the host, but it impairs the colonization and proliferation in vascular tissues. The ΔVdAsp1 mutant shows defective initiation of conidiophore formation and reduced expression of genes associated with the central developmental pathway. By live-cell imaging, we observed that some of ΔVdAsp1 mutant hyphae are swollen, and microtubule arrangements at the apical region of these hyphae are disorganized. These results indicate that VdAsp1 regulates the transition from vegetative growth to asexual reproduction by modulating microtubule dynamic organization, which is essential for V. dahliae to colonize and proliferate in vascular tissues. These findings provided a potential new direction in the control of vascular wilt pathogen by targeting conidium production in vascular tissues.

Three-Dimensional Printed Poly (Lactic-co-Glycolic Acid)-Magnesium Composite Scaffolds for the Promotion of Osteogenesis Through Immunoregulation.

Scaffolds play an important role in bone tissue engineering. The ideal engineered scaffold needs to be biocompatible, bioactive, and able to regulate immune cells to enhance bone regeneration. In this study, magnesium (Mg)-contained poly(lactic-co-glycolic acid) (PLGA) scaffolds (hereinafter, referred to as PLGA-2Mg) were fabricated by 3-dimensional printing using a mixture of PLGA and MgSO 4 powder. Poly(lactic-co-glycolic acid) scaffolds (hereinafter, referred to as PLGA) were also fabricated by 3-dimensional printing and were used as control. The biocompatibility, immunoregulatory ability, and osteogenic properties of PLGA-2Mg were analyzed and compared with those of PLGA. The results indicate that the incorporation of Mg increased the Young modulus and surface roughness of the scaffold, but did not affect its degradation. The PLGA-2Mg further promoted the adhesion and proliferation of MC3T3-E1 cells compared with PLGA, which indicates its improved biocompatibility and bioactivity. In addition, PLGA-2Mg inhibited the polarization of RAW 264.7 cells toward the M1 phenotype by down-regulating the IL-1ß , IL-6 , and iNOs gene expression when challenged with lipopolysaccharide stimulation. In contrast, it promoted the polarization of RAW 264.7 cells toward the M2 phenotype by up-regulating the TGF-ß , IL-10 , and Arg-1 gene expression without lipopolysaccharide stimulation. Finally, MC3T3-E1 cells were cocultured with RAW 264.7 cells and scaffolds using a transwell system. It was found that the expression level of osteogenic-related genes ( ALP , COL-1 , BMP2 , and BSP ) was significantly upregulated in the PLGA-2Mg group compared with that in the PLGA group. Consequently, PLGA-2Mg with increased biocompatibility and bioactivity can promote osteogenesis through immunoregulation and has the potential to be used as a novel scaffold in bone tissue engineering.

A Novel Prediction Method of Transfer-Assisted Action Oriented to Individual Differences for the Excretion Care Robot.

The excretion care robot's (ECR) accurate recognition of transfer-assisted actions is crucial during its usage. However, transfer action recognition is a challenging task, especially since the differentiation of actions seriously affects its recognition speed, robustness, and generalization ability. We propose a novel approach for transfer action recognition assisted by a bidirectional long- and short-term memory (Bi-LSTM) network combined with a multi-head attention mechanism. Firstly, we utilize posture sensors to detect human movements and establish a lightweight three-dimensional (3D) model of the lower limbs. In particular, we adopt a discrete extended Kalman filter (DEKF) to improve the accuracy and foresight of pose solving. Then, we construct an action prediction model that incorporates a fused Bi-LSTM with Multi-head attention (MHA Bi-LSTM). The MHA extracts key information related to differentiated movements from different dimensions and assigns varying weights. Utilizing the Bi-LSTM network effectively combines past and future information to enhance the prediction results of differentiated actions. Finally, comparisons were made by three subjects in the proposed method and with two other time series based neural network models. The reliability of the MHA Bi-LSTM method was verified. These experimental results show that the introduced MHA Bi-LSTM model has a higher accuracy in predicting posture sensor-based excretory care actions. Our method provides a promising approach for handling transfer-assisted action individual differentiation in excretion care tasks.

Efficient Near-Infrared Electroluminescence from Lanthanide-Doped Perovskite Quantum Cutters.

Perovskite nanocrystals (PeNCs) deliver size- and composition-tunable luminescence of high efficiency and color purity in the visible range. However, attaining efficient electroluminescence (EL) in the near-infrared (NIR) region from PeNCs is challenging, limiting their potential applications. Here we demonstrate a highly efficient NIR light-emitting diode (LED) by doping ytterbium ions into a PeNCs host (Yb3+ : PeNCs), extending the EL wavelengths toward 1000 nm, which is achieved through a direct sensitization of Yb3+ ions by the PeNC host. Efficient quantum-cutting processes enable high photoluminescence quantum yields (PLQYs) of up to 126 % from the Yb3+ : PeNCs. Through halide-composition engineering and surface passivation to improve both PLQY and charge-transport balance, we demonstrate an efficient NIR LED with a peak external quantum efficiency of 7.7 % at a central wavelength of 990 nm, representing the most efficient perovskite-based LEDs with emission wavelengths beyond 850 nm.

Distribution of Diatoms in the Navigable Sections of Beijing-Hangzhou Grand Canal.

OBJECTIVES: To establish a diatom database by analyzing the quatity, species distribution and differences of diatom in water samples of the whole navigable sections of the Beijing-Hangzhou Grand Canal, to provide a reference for the inference of the drowning site. METHODS: Water samples were collected at 22 sites in the navigable sections of the Beijing-Hangzhou Grand Canal (Jining section to Yangzhou Section), and the diatoms at each site were qualitatively and quantitatively analyzed by using graphite digestion-scanning electron microscopy. RESULTS: Sampling site T (Laohuaijiang River Line, Gaoyou City, Yangzhou City, Jiangsu Province) had the highest number of diatoms, while sampling site O (Siyang County, Suqian City, Jiangsu Province) had the lowest number of diatoms, with a large gap of 68 times. At sampling site Q (Jiangpu District, Huaian city, Jiangsu Province), there were 19 species of diatoms. The sampling site O had the least diatoms, with 7 species. There were no significant differences in species evenness and species diversity at each sampling site (P>0.05). Some sampling sites have characterized diatoms, such as Caloneis at station A (Taibai Lake, Weishan County, Shandong Province), Rhoicosphenia at station B (Nanyang Town, Weishan County, Shandong Province), Amphora at station I (Taierzhuang District, Zaozhuang City, Shandong Province) and Epithemia at station J (Pizhou 310 national highway, Xuzhou City, Jiangsu Province). CONCLUSIONS: The species richness of diatoms gradually increased from north to south. Diatom species richness and species diversity might be higher in areas with complex environments and large population flow. Climate type has a certain influence on the distribution of diatoms.

KTN80 confers precision to microtubule severing by specific targeting of katanin complexes in plant cells.

The microtubule (MT)-severing enzyme katanin triggers dynamic reorientation of cortical MT arrays that play crucial functions during plant cell morphogenesis, such as cell elongation, cell wall biosynthesis, and hormonal signaling. MT severing specifically occurs at crossover or branching nucleation sites in living Arabidopsis cells. This differs from the random severing observed along the entire length of single MTs in vitro and strongly suggests that a precise control mechanism must exist in vivo However, how katanin senses and cleaves at MT crossover and branching nucleation sites in vivo has remained unknown. Here, we show that the katanin p80 subunit KTN80 confers precision to MT severing by specific targeting of the katanin p60 subunit KTN1 to MT cleavage sites and that KTN1 is required for oligomerization of functional KTN80-KTN1 complexes that catalyze severing. Moreover, our findings suggest that the katanin complex in Arabidopsis is composed of a hexamer of KTN1-KTN80 heterodimers that sense MT geometry to confer precise MT severing. Our findings shed light on the precise control mechanism of MT severing in plant cells, which may be relevant for other eukaryotes.

Tumor mutation burden in connection with immune-related survival in uterine corpus endometrial carcinoma.

BACKGROUND: UCEC is the most common gynecological malignancy in many countries, and its mechanism of occurrence and development is related to tumor mutation burden (TMB) and immune cell infiltration. Therefore, it is necessary to systematically explore the TMB-related gene profile in immune cells to improve the prognosis of UCEC. METHODS: We integrated TMB-related genes with basic clinical information of UCEC patients based on TCGA dataset. Differentially expressed genes (DEGs) were selected through differential expression screening, PPI, and enrichment analysis. Additionally, we analyzed the components of immune cell infiltration of the DEGs to obtain the differential immunity-related genes. A single factor and multifactor Cox regression analyses were conducted to establish new prognostic indicators of OS and DFS based on TMB-related immune genes. To further study the correlation between survival and immune cell infiltration, a Cox model based on these immune infiltration compositions was built. Using the clinical variables, we established nomograms for OS and DFS. RESULTS: 393 DEGs were significantly associated with clinical outcomes and the immune component in patients with UCEC. Gene Ontology (GO) and Kyoto Encyclopedia of Genes, Genomes (KEGG) pathway and protein-protein interaction network (PPI) analyses revealed the role of these genes and information on related pathways. Then, two prognostic models were established based on the differential immune genes for OS (GFAP and MX2) and DFS (MX2, GFAP, IGHM, FGF20, and TRAV21). In DFS, the differential immune genes were related to CD4+ T cell, CD8+ T cell, macrophage, and neutrophil (all P < 0.05). B cell and CD8+ T cell were independent prognostic factors from among the immune cell elements in UCEC. Finally, the risk scores of these models were combined with the clinical elements-based nomogram models, and the AUC values were all over 0.7. CONCLUSIONS: Our results identified several clinically significant differential immune genes and established relevant prognostic models, providing a basis for the molecular analysis of TMB and immune cells in UCEC, and identified potential prognostic and immune-related genes for UCEC. We added clinical related conditions for further analysis to confirm the identity of the genes and clinical elements-based models.

A Modified Actin (Gly65Val Substitution) Expressed in Cotton Disrupts Polymerization of Actin Filaments Leading to the Phenotype of Ligon Lintless-1 (Li1) Mutant.

Dynamic remodeling of the actin cytoskeleton plays a central role in the elongation of cotton fibers, which are the most important natural fibers in the global textile industry. Here, a high-resolution mapping approach combined with comparative sequencing and a transgenic method revealed that a G65V substitution in the cotton actin Gh_D04G0865 (GhACT17D in the wild-type) is responsible for the Gossypium hirsutum Ligon lintless-1 (Li1) mutant (GhACT17DM). In the mutant GhACT17DM from Li1 plant, Gly65 is substituted with valine on the lip of the nucleotide-binding domain of GhACT17D, which probably affects the polymerization of F-actin. Over-expression of GhACT17DM, but not GhACT17D, driven by either a CaMV35 promoter or a fiber-specific promoter in cotton produced a Li1-like phenotype. Compared with the wild-type control, actin filaments in Li1 fibers showed higher growth and shrinkage rates, decreased filament skewness and parallelness, and increased filament density. Taken together, our results indicate that the incorporation of GhACT17DM during actin polymerization disrupts the establishment and dynamics of the actin cytoskeleton, resulting in defective fiber elongation and the overall dwarf and twisted phenotype of the Li1 mutant.

The host actin cytoskeleton channels rhizobia release and facilitates symbiosome accommodation during nodulation in Medicago truncatula.

In plants, the actin cytoskeleton plays a central role in regulating intracellular transport and trafficking in the endomembrane system. Work in legumes suggested that during nodulation, the actin cytoskeleton coordinates numerous cellular processes in the development of nitrogen-fixing nodules. However, we lacked live-cell visualizations demonstrating dynamic remodeling of the actin cytoskeleton during infection droplet release and symbiosome development. Here, we generated transgenic Medicago truncatula lines stably expressing the fluorescent actin marker ABD2-GFP, and utilized live-cell imaging to reveal the architecture and dynamics of the actin cytoskeleton during nodule development. Live-cell observations showed that different zones in nitrogen-fixing nodules exhibit distinct actin architectures and infected cells display five characteristic actin architectures during nodule development. Live-cell imaging combined with three-dimensional reconstruction demonstrated that dense filamentous-actin (F-actin) arrays channel the elongation of infection threads and the release of infection droplets, an F-actin network encircles freshly-released rhizobia, and short F-actin fragments and actin dots around radially distributed symbiosomes. Our findings suggest an important role of the actin cytoskeleton in infection droplet release, symbiosome development and maturation, and provide significant insight into the cellular mechanisms underlying nodule development and nitrogen fixation during legume-rhizobia interactions.

Lymphadenectomy and prognosis for elderly females with stage I endometrioid endometrial cancer.

PURPOSE: The potential therapeutic benefits of lymphadenectomy in endometrial cancer (EC) patients are still ambiguous. Therefore, a population-based retrospective analysis was conducted to determine the association between lymphadenectomy and survival in elderly female patients with stage I endometrioid EC. METHODS: The Surveillance, Epidemiology, and End Results (SEER) program database was retrospectively analyzed, and data of 63,372 female patients with early-stage type I EC from 1988 to 2013 were collected. The main patient and tumor characteristics included marital status, age, ethnicity, time of diagnosis, tumor grade, radiotherapy, and lymphadenectomy status. Kaplan-Meier and Cox proportional hazard regression analyses were performed to determine the association between lymph node dissection and the overall survival (OS) and cancer-specific survival in women older than 50 years with stage I endometrioid EC. RESULTS: The majority (83.7%) of the patients who met the inclusion criteria for the study were older than 50 years. In both grade 1 and 2 patients aged over 50 years, lymph node conservation was associated with a higher mortality risk compared to lymphadenectomy (all P < 0.005). Multivariate analysis indicated that lymphadenectomy was an independent predictor of improved OS in early-stage type 1 EC patients, with hazard ratios of 0.893 and 0.827 for the grade 1 and grade 2 patients, respectively (P < 0.0001). CONCLUSIONS: Lymphadenectomy could improve long-term OS in women older than 50 years with grade 1 and 2 endometrioid EC.

Towards a better recording of microtubule cytoskeletal spatial organization and dynamics in plant cells.

Numerous fluorescent marker lines are currently available to visualize microtubule (MT) architecture and dynamics in living plant cells, such as markers expressing p35S::GFP-MBD or p35S::GFP-TUB6. However, these MT marker lines display obvious defects that affect plant growth or produce unstable fluorescent signals. Here, a series of new marker lines were developed, including the pTUB6::VisGreen-TUB6-expressing line in which TUB6 is under the control of its endogenous regulatory elements and eGFP is replaced with VisGreen, a brighter fluorescent protein. Moreover, two different markers were combined into one expression vector and developed two dual-marker lines. These marker lines produce bright, stable fluorescent signals in various tissues, and greatly shorten the screening process for generating dual-marker lines. These new marker lines provide a novel resource for MT research.

Superior Catalytic Performance of Gold Nanoparticles Within Small Cross-Linked Lysozyme Crystals.

Bionanomaterials synthesized by bioinspired templating methods have emerged as a novel class of composite materials with varied applications in catalysis, detection, drug delivery, and biomedicine. In this study, two kinds of cross-linked lysozyme crystals (CLLCs) with different sizes were applied for the in situ growth of Au nanoparticles (AuNPs). The resulting composite materials were characterized by light microscopy, scanning electron microscopy, transmission electron microscopy, thermogravimetric analysis, and X-ray photoelectron spectroscopy. The catalytic properties of the prepared materials were examined in the catalytic reduction of 4-nitrophenol (4-NP) to 4-aminophenol (4-AP). It was found that the size of the AuNPs increased with an increase in Au loading for both small and large crystals. In addition, small crystals favored homogeneous adsorption and distribution of the metal precursors. And the size of the AuNPs within small crystals could be maintained below 2.5 nm by managing the HAuCl4/lysozyme molar ratio. Furthermore, the lysozyme functional groups blocked the AuNP activity sites, therefore reducing their catalytic activity. This effect was more pronounced for small AuNPs. Moreover, the mass transfer of reactants (4-NP) from solution to AuNPs within the crystals restricted their catalytic reduction, leading to superior catalytic performance of the AuNPs within small cross-linked lysozyme crystals (Au@S-CLLCs) compared to those within large cross-linked lysozyme crystals (Au@L-CLLCs) at similar Au loadings. Finally, an increase in Au loading clogged the crystal channels with increased quantities of larger AuNPs, thus impeding the catalytic performance of Au@S-CLLCs.

Prediction of dose distributions for non-small cell lung cancer patients using MHA-ResUNet.

BACKGROUND: The current level of automation in the production of radiotherapy plans for lung cancer patients is relatively low. With the development of artificial intelligence, it has become a reality to use neural networks to predict dose distributions and provide assistance for radiation therapy planning. However, due to the significant individual variability in the distribution of non-small cell lung cancer (NSCLC) planning target volume (PTV) and the complex spatial relationships between the PTV and organs at risk (OARs), there is still a lack of a high-precision dose prediction network tailored to the characteristics of NSCLC. PURPOSE: To assist in the development of volumetric modulated arc therapy (VMAT) plans for non-small cell lung cancer patients, a deep neural network is proposed to predict high-precision dose distribution. METHODS: This study has developed a network called MHA-ResUNet, which combines a large-kernel dilated convolution module and multi-head attention (MHA) modules. The network was trained based on 80 VMAT plans of NSCLC patients. CT images, PTV, and OARs were fed into the independent input channel. The dose distribution was taken as the output to train the model. The performance of this network was compared with that of several commonly used networks, and the networks' performance was evaluated based on the voxel-level mean absolute error (MAE) within the PTV and OARs, as well as the error in clinical dose-volume metrics. RESULTS: The MAE between the predicted dose distribution and the manually planned dose distribution within the PTV is 1.43 Gy, and the D95 error is less than 1 Gy. Compared with the other three commonly used networks, the dose error of the MHA-ResUNet is the smallest in PTV and OARs. CONCLUSIONS: The proposed MHA-ResUNet network improves the receptive field and filters the shallow features to learn the relative spatial relation between the PTV and the OARs, enabling accurate prediction of dose distributions in NSCLC patients undergoing VMAT radiotherapy.

Cobalt etched graphite felt electrode for enhanced removal of organic pollutant in aqueous solution with a solid polymer electrolyte.

In this study, cobalt etched graphite felt electrodes were produced using a simple etching technique. It was used in combination with a solid polymer electrolyte (SPE) for the degradation of the target contaminant Orange II by Electro-Fenton (EF) technique in low conductivity water. In this method, 94% of Orange II in low conductivity water was removed in 90 min. The characterization analysis substantiates the hypothesis that the electrodes produced exhibit a three-dimensional porous structure, augmented defect concentration, and enhanced electron transfer capability. In addition, the potential reaction mechanism was inferred from the radical quenching experiments, and hydroxyl radicals (·OH) were deemed the main reactive substances. The combination of cobalt etched graphite felt electrodes with SPE demonstrates remarkable efficacy in the treatment of organic wastewater characterized by low electrical conductivity.

Lateral epitaxial growth of two-dimensional organic heterostructures.

Two-dimensional organic lateral heterostructures (2D OLHs) are attractive for the fabrication of functional materials. However, it is difficult to control the nucleation, growth and orientation of two distinct components. Here we report the combination of two methods-liquid-phase growth and vapour-phase growth-to synthesize 2D OLHs from perylene and a perylenecarboxaldehyde derivative, with a lateral size of ~20 µm and a tunable thickness ranging from 20 to 400 nm. The screw dislocation growth behaviour of the 2D crystals shows the spiral arrangement of atoms within the crystal lattice, which avoids volume expansion and contraction of OLH, thereby minimizing lateral connection defects. Selective control of the nucleation and sequential growth of 2D crystals leads to structural inversion of the 2D OLHs by the vapour-phase growth method. The resulting OLHs show good light-transport capabilities and tunable spatial exciton conversion, useful for photonic applications. This synthetic strategy can be extended to other families of organic polycyclic aromatic hydrocarbons, as demonstrated with other pyrene and perylene derivatives.

Cross-linked lysozyme crystal templated synthesis of Au nanoparticles as high-performance recyclable catalysts.

Bio-nanomaterials fabricated using a bioinspired templating technique represent a novel class of composite materials with diverse applications in biomedical, electronic devices, drug delivery, and catalysis. In this study, Au nanoparticles (NPs) are synthesized within the solvent channels of cross-linked lysozyme crystals (CLLCs) in situ without the introduction of extra chemical reagents or physical treatments. The as-prepared AuNPs-in-protein crystal hybrid materials are characterized by light microscopy, transmission electron microscopy, x-ray diffraction, and Fourier-transform infrared spectroscopy analyses. Small AuNPs with narrow size distribution reveal the restriction effects of the porous structure in the lysozyme crystals. These composite materials are proven to be active heterogeneous catalysts for the reduction of 4-nitrophenol to 4-aminophenol. These catalysts can be easily recovered and reused at least 20 times because of the physical stability and macro-dimension of CLLCs. This work is the first to use CLLCs as a solid biotemplate for the preparation of recyclable high-performance catalysts.

Shape evolution and thermal stability of lysozyme crystals: effect of pH and temperature.

The properties of crystalline protein materials are closely linked to crystal shape. However, the effective strategies for the shape control of protein crystals are lacking. The conventional sitting-drop vapor-diffusion method was employed to investigate the influence of pH and temperature on the crystal nucleation behavior of hen egg white lysozyme. Moreover, the size distributions of protein crystals grown at different conditions were analyzed. Differential scanning calorimetry was employed to evaluate the thermal stability of lysozyme crystals. The results indicated that pH and temperature will affect the supersaturation and electrostatic interactions among protein molecules in the nucleation process. In particular, the crystals with different aspect ratios can be selectively nucleated, depending upon the choice of pH and temperature. Therefore, this study provided a simple method for obtaining shape-controlled lysozyme crystals and supplied some information on thermal behaviors of lysozyme crystals grown at different pH values.

Visualization of Cytoskeleton Organization and Dynamics in Elongating Cotton Fibers by Live-Cell Imaging.

Cotton fibers are extremely elongated single cells and have long been regarded as an ideal model to investigate polarized plant cell elongation. Actin filaments (F-actin), as well as the cortical microtubules (CMTs), play vital roles in polarized cell growth and morphogenesis. We have generated stable transgenic cotton plants expressing fluorescent markers for the actin and microtubule cytoskeletons. Further live-cell imaging identified dynamic features of the F-actin and cortical microtubule (CMT) architectures and discovered that cotton fibers elongate in a unique tip-biased diffuse growth mode. Here, we describe methods for preparing growing cotton fiber samples, as well as the visualization of cytoskeletal organization and dynamics by live-cell imaging. Combined with comprehensive image analyses, these methods can be used to identify how cytoskeleton organization and dynamics determine cell morphogenesis in highly polarized cotton fibers.