Heterozygous variants in USP25 cause genetic generalized epilepsy.

USP25 encodes ubiquitin-specific proteases 25, a key member of deubiquitinating enzyme family and is involved in neural fate determination. Although abnormal expression in Down's syndrome was reported previously, the specific role of USP25 in human diseases has not been defined. In this study, we performed trio-based whole exome sequencing in a cohort of 319 cases (families) with generalized epilepsy of unknown etiology. Five heterozygous USP25 variants including two de novo and three co-segregated variants were determined in eight individuals affected by generalized seizures and/or febrile seizures from five unrelated families. The frequency of USP25 variants showed a significantly high aggregation in this cohort compared to the East Asian population and all populations in the gnomAD database. The mean onset ages of febrile and afebrile seizures were 10 months (infancy) and 11.8 years (juvenile), respectively. The patients achieved seizure freedom except one had occasional nocturnal seizures at the last follow-up. Two patients exhibited intellectual disability. Usp25 was ubiquitously expressed in mouse brain with two peaks on embryonic days (E14‒E16) and postnatal day 21, respectively. Similarly, USP25 expressed in fetus/early childhood stage with a second peak at approximately 12‒20 years old in human brain, consistent with the seizure onset age at infancy and juvenile in the patients. To investigate the functional impact of USP25 deficiency in vivo, we established Usp25 knock-out mice, which showed increased seizure susceptibility compared to wild-type mice in pentylenetetrazol-induced seizure test. To explore the impact of USP25 variants, we employed multiple functional detections. In HEK293T cells, the severe phenotype associated variant (p.Gln889Ter) led to a significant reduction of mRNA and protein expressions but formed a stable truncated dimers with increment of deubiquitinating enzyme activities and abnormal cellular aggregations, indicating a gain-of-function effect. The p.Gln889Ter and p.Leu1045del increased neuronal excitability in mice brain, with a higher firing ability in p.Gln889Ter. These functional impairments align with the severity of the observed phenotypes, suggesting a genotype-phenotype correlation. Hence, a moderate association between USP25 and epilepsy was noted, indicating USP25 is potentially a predisposing gene for epilepsy. Our results from Usp25 null mice and the patient-derived variants indicated that USP25 would play epileptogenic role via loss-of-function or gain-of-function effects. The truncated variant p.Gln889Ter would have profoundly different effect on epilepsy. Together, our results underscore the significance of USP25 heterozygous variants in epilepsy, thereby highlighting the critical role of USP25 in the brain.

Exploiting Topological Darkness in Photonic Crystal Slabs for Spatiotemporal Vortex Generation.

Spatiotemporal optical vortices (STOVs) with swirling phase singularities in space and time hold great promise for a wide range of applications across diverse fields. However, current approaches to generate STOVs lack integrability and rely on bulky free-space optical components. Here, we demonstrate routine STOV generation by harnessing the topological darkness phenomenon of a photonic crystal slab. Complete polarization conversion enforced by symmetry enables topological darkness to arise from photonic bands of guided resonances, imprinting vortex singularities onto an ultrashort reflected pulse. Utilizing time-resolved spatial mapping, we provide the first observation of STOV generation using a photonic crystal slab, revealing the imprinted STOV structure manifested as a curved vortex line in the pulse profile in space and time. Our work establishes photonic crystal slabs as a versatile and accessible platform for engineering STOVs and harnessing the topological darkness in nanophotonics.

Self-Limiting Phase Transition Enabling Reversible Overstoichiometric Li Storage in Ni-Rich Cathodes.

Ni-rich cathodes are some of the most promising candidates for advanced lithium-ion batteries, but their available capacities have been stagnant due to the intrinsic Li+ storage sites. Extending the voltage window down can induce the phase transition from O3 to 1T of LiNiO2-derived cathodes to accommodate excess Li+ and dramatically increase the capacity. By setting the discharge cutoff voltage of LiNi0.6Co0.2Mn0.2O2 to 1.4 V, we can reach an extremely high capacity of 393 mAh g-1 and an energy density of 1070 Wh kg-1 here. However, the phase transition causes fast capacity decay and related structural evolution is rarely understood, hindering the utilization of this feature. We find that the overlithiated phase transition is self-limiting, which will transform into solid-solution reaction with cycling and make the cathode degradation slow down. This is attributed to the migration of abundant transition metal ions into lithium layers induced by the overlithiation, allowing the intercalation of overstoichiometric Li+ into the crystal without the O3 framework change. Based on this, the wide-potential cycling stability is further improved via a facile charge-discharge protocol. This work provides deep insight into the overstoichiometric Li+ storage behaviors in conventional layered cathodes and opens a new avenue toward high-energy batteries.

Generation of Spatiotemporal Vortex Pulses by Resonant Diffractive Grating.

Spatiotemporal vortex pulses are wave packets that carry transverse orbital angular momentum, exhibiting exotic structured wave fronts that can twist through space and time. Existing methods to generate these pulses require complex setups like spatial light modulators or computer-optimized structures. Here, we demonstrate a new approach to generate spatiotemporal vortex pulses using just a simple diffractive grating. The key is constructing a phase vortex in frequency-momentum space by leveraging symmetry, resonance, and diffraction. Our approach is applicable to any wave system. We use a liquid surface wave (gravity wave) platform to directly demonstrate and observe the real-time generation and evolution of spatiotemporal vortex pulses. This straightforward technique provides opportunities to explore pulse dynamics and potential applications across different disciplines.

Stimulus-responsive proteins involved in multi-process regulation of storage substance accumulation during rice grain filling under elevated temperature.

BACKGROUND: The intensified global warming during grain filling deteriorated rice quality, in particular increasing the frequency of chalky grains which markedly impact market value. The formation of rice quality is a complex process influenced by multiple genes, proteins and physiological metabolic processes. Proteins responsive to stimulus can adjust the ability of plants to respond to unfavorable environments, which may be an important protein involved in the regulation of quality formation under elevated temperature. However, relatively few studies have hindered our further understanding of rice quality formation under elevated temperature. RESULTS: We conducted the actual field elevated temperature experiment and performed proteomic analysis of rice grains at the early stage of grain filling. Starting with the response to stimulus in GO annotation, 22 key proteins responsive to stimulus were identified in the regulation of grain filling and response to elevated temperature. Among the proteins responsive to stimulus, during grain filling, an increased abundance of signal transduction and other stress response proteins, a decreased abundance of reactive oxygen species-related proteins, and an increased accumulation of storage substance metabolism proteins consistently contributed to grain filling. However, the abundance of probable indole-3-acetic acid-amido synthetase GH3.4, probable indole-3-acetic acid-amido synthetase GH3.8 and CBL-interacting protein kinase 9 belonged to signal transduction were inhibited under elevated temperature. In the reactive oxygen species-related protein, elevated temperature increased the accumulation of cationic peroxidase SPC4 and persulfide dioxygenase ETHE1 homolog to maintain normal physiological homeostasis. The increased abundance of alpha-amylase isozyme 3E and seed allergy protein RA5 was related to the storage substance metabolism, which regulated starch and protein accumulation under elevated temperature. CONCLUSION: Auxin synthesis and calcium signal associated with signal transduction, other stress responses, protein transport and modification, and reactive oxygen species-related proteins may be key proteins responsive to stimulus in response to elevated temperature. Alpha-amylase isozyme 3E and seed allergy protein RA5 may be the key proteins to regulate grain storage substance accumulation and further influence quality under elevated temperature. This study enriched the regulatory factors involved in the response to elevated temperature and provided a new idea for a better understanding of grain response to temperature.

Realization of large transmitted Goos-Hänchen shifts with high (near 100%) transmittance based on a coupled double-layer grating system.

Achieving Goos-Hänchen shift enhancement with high transmittance or reflectance based on the resonance effect is challenging due to the drop in the resonance region. This Letter demonstrates the realization of large transmitted Goos-Hänchen shifts with high (near 100%) transmittance based on a coupled double-layer grating system. The double-layer grating is composed of two parallel and misaligned subwavelength dielectric gratings. By changing the distance and the relative dislocation between the two dielectric gratings, the coupling of the double-layer grating can be flexibly tuned. The transmittance of the double-layer grating can be close to 1 in the entire resonance angle region, and the gradient of the transmissive phase is also preserved. The Goos-Hänchen shift of the double-layer grating reaches ∼30 times the wavelength, approaching 1.3 times the radius of the beam waist, which can be observed directly.

Chemical-Mechanical Robustness of Single-Crystalline Ni-Rich Cathode Enabled by Surface Atomic Arrangement Control.

Layered transition metal oxide cathodes have been one of the dominant cathodes for lithium-ion batteries with efficient Li+ intercalation chemistry. However, limited by the weak layered interaction and unstable surface, mechanical and chemical failure plagues their electrochemical performance, especially for Ni-rich cathodes. Here, adopting a simultaneous elemental-structural atomic arrangement control based on the intrinsic Ni-Co-Mn system, the surface role is intensively investigated. Within the invariant oxygen sublattice of the crystal, a robust surface with the synergistic concentration gradient and layered-spinel intertwined structure is constructed on the model single-crystalline Ni-rich cathode. With mechanical strain dissipation and chemical erosion suppression, the cathode exhibits an impressive capacity retention of 82 % even at the harsh 60 °C after 150 cycles at 1 C. This work highlights the coupling effect of structure and composition on the chemical-mechanical properties, and the concept will spur more researches on the cathodes that share the same sublattice.

Crystal idioblasts are involved in the anther dehiscence of Nicotiana tabacum.

In Nicotiana tabacum, the degeneration of connective tissue and stomium tissue (the stomium and circular cell cluster [CCC]) is essential for anther dehiscence. Both connective cells and CCC cells are crystal idioblasts, and these cells will undergo degeneration after accumulating calcium oxalate (CaOx) crystals. However, detailed data concerning this process are minimal. Therefore, this study used cellular biological and physiological methods to illustrate this relationship. Results demonstrated that tobacco anther dehiscence is a series of timed programmed cell death (PCD) processes that include the CCC, connective tissue, and stomium. The degenerating crystal idioblasts of the tobacco anther were found to possess two hallmark characteristics that distinguished them from normal PCD cells, namely dynamic changes in CaOx crystals and the appearance of numerous peroxisomes. The accumulation of CaOx and the production of H2 O2 occurred simultaneously or successively before PCD. The peak H2 O2 content was found to appear after the insoluble oxalate. Further, CeCl3 cytochemistry staining was used to detect subcellular H2 O2 , and the precipitate of H2 O2 was primarily present in peroxisomes and around CaOx crystals. These results show that anther dehiscence in N. tabacum is a PCD process in which crystal idioblasts play a vital role in CaOx degradation and H2 O2 production.

Low-latency monaural speech enhancement with deep filter-bank equalizer.

It is highly desirable that speech enhancement algorithms can achieve good performance while keeping low latency for many applications, such as digital hearing aids, mobile phones, acoustically transparent hearing devices, and public address systems. To improve the performance of traditional low-latency speech enhancement algorithms, a deep filter-bank equalizer (FBE) framework was proposed that integrated a deep learning-based subband noise reduction network with a deep learning-based shortened digital filter mapping network. In the first network, a deep learning model was trained with a controllable small frame shift to satisfy the low-latency demand, i.e., no greater than 4 ms, so as to obtain (complex) subband gains that could be regarded as an adaptive digital filter in each frame. In the second network, to reduce the latency, this adaptive digital filter was implicitly shortened by a deep learning-based framework and was then applied to noisy speech to reconstruct the enhanced speech without the overlap-add method. Experimental results on the WSJ0-SI84 corpus indicated that the proposed DeepFBE with only 4-ms latency achieved much better performance than traditional low-latency speech enhancement algorithms across several objective metrics. Listening test results further confirmed that our approach achieved higher speech quality than other methods.

Polarization Singularities of Photonic Quasicrystals in Momentum Space.

We report the observation of polarization singularities in momentum space of 2D photonic quasicrystal slabs. Supercell approximation and band-unfolding approach are applied to obtain approximate photonic dispersions and the far-field polarization states defined on them. We discuss the relations between the topological charges of the polarization vortex singularities at Γ points and the symmetries of photonic quasicrystal slabs. With a perspective of multipolar expansions for the supercell, we confirm that the singularities are protected by the point-group symmetry of the photonic quasicrystal slab. We further uncover that the polarization singularities of photonic quasicrystal slab correspond to quasibound states in the continuum with exceptionally high-quality factors. Polarization singularities of different topological charges are also experimentally verified. Our Letter introduces core concepts of optical singularities into quasiperiodic systems, providing new platforms for explorations merging topological and singular optics.

Binary Neural Network for Automated Visual Surface Defect Detection.

As is well-known, defects precisely affect the lives and functions of the machines in which they occur, and even cause potentially catastrophic casualties. Therefore, quality assessment before mounting is an indispensable requirement for factories. Apart from the recognition accuracy, current networks suffer from excessive computing complexity, making it of great difficulty to deploy in the manufacturing process. To address these issues, this paper introduces binary networks into the area of surface defect detection for the first time, for the reason that binary networks prohibitively constrain weight and activation to +1 and -1. The proposed Bi-ShuffleNet and U-BiNet utilize binary convolution layers and activations in low bitwidth, in order to reach comparable performances while incurring much less computational cost. Extensive experiments are conducted on real-life NEU and Magnetic Tile datasets, revealing the least OPs required and little accuracy decline. When classifying the defects, Bi-ShuffleNet yields comparable results to counterpart networks, with at least 2× inference complexity reduction. Defect segmentation results indicate similar observations. Some network design rules in defect detection and binary networks are also summarized in this paper.

Dissecting MicroRNA-mRNA Regulatory Networks Underlying Sulfur Assimilation and Cadmium Accumulation in Poplar Leaves.

The process of cadmium (Cd) accumulation and detoxification under different sulfur levels remains largely unknown in woody plants. To investigate the physiological and transcriptomic regulation mechanisms of poplars in response to different sulfate (S) supply levels and Cd exposure, we exposed Populus deltoides saplings to one of the low, moderate and high S levels together with either 0 or 50 µM Cd. Cd accumulation was decreased in low S-treated poplar leaves, and it tended to be increased in high S-supplied leaves under the Cd exposure condition. Sulfur nutrition was deficient in low S-supplied poplars, and it was improved in high S-treated leaves. Cd exposure resulted in lower sulfur level in the leaves supplied with moderate S, it exacerbated a Cd-induced sulfur decrease in low S-treated leaves and it caused a higher sulfur concentration in high S-supplied leaves. In line with the physiological changes, a number of mRNAs and microRNAs (miRNAs) involved in Cd accumulation and sulfur assimilation were identified and the miRNA-mRNA networks were dissected. In the networks, miR395 and miR399 members were identified as hub miRNAs and their targets were ATP sulfurylase 3 (ATPS3) and phosphate 2 (PHO2), respectively. These results suggest that Cd accumulation and sulfur assimilation are constrained by low and enhanced by high S supply, and Cd toxicity is aggravated by low and relieved by high S in poplar leaves, and that miRNA-mRNA regulatory networks play pivotal roles in sulfur-mediated Cd accumulation and detoxification in Cd-exposed poplars.

Competing Endogenous RNA Networks Underlying Anatomical and Physiological Characteristics of Poplar Wood in Acclimation to Low Nitrogen Availability.

Although poplar plantations are often established on nitrogen (N)-poor soil, the physiological and molecular mechanisms underlying wood properties of poplars in acclimation to low N availability remain largely unknown. To investigate wood properties of poplars in acclimation to low N, Populus � canescens saplings were exposed to either 50 (low N) or 500 (normal N) �M NH4NO3 for 2 months. Low N resulted in decreased xylem width and cell layers of the xylem (the number of cells counted along the ray parenchyma on the stem cross section), narrower lumina of vessels and fibers, greater thickness of double fiber walls (the walls between two adjacent fiber cells), more hemicellulose and lignin deposition, and reduced cellulose accumulation in poplar wood. Consistently, concentrations of gibberellins involved in cell size determination and the abundance of various metabolites including amino acids, carbohydrates and precursors for cell wall biosynthesis were decreased in low N-supplied wood. In line with these anatomical and physiological changes, a number of mRNAs, long noncoding RNAs (lncRNAs) and microRNAs (miRNAs) were significantly differentially expressed. Competing endogenous RNA regulatory networks were identified in the wood of low N-treated poplars. Overall, these results indicate that miRNAs-lncRNAs-mRNAs networks are involved in regulating wood properties and physiological processes of poplars in acclimation to low N availability.

Anatomy and RNA-Seq reveal important gene pathways regulating sex differentiation in a functionally Androdioecious tree, Tapiscia sinensis.

BACKGROUND: Gametogenesis is a key step in the production of ovules or pollen in higher plants. The sex-determination aspects of gametogenesis have been well characterized in the model plant Arabidopsis. However, little is known about this process in androdioecious plants. Tapiscia sinensis Oliv. is a functionally androdioecious tree, with both male and hermaphroditic individuals. Hermaphroditic flowers (HFs) are female-fertile flowers that can produce functional pollen and set fruits. However, compared with male flowers (MFs), the pollen viability and number of pollen grains per flower are markedly reduced in HFs. MFs are female-sterile flowers that fail to set fruit and that eventually drop. RESULTS: Compared with HF, a notable cause of MF female sterility in T. sinensis is when the early gynoecium meristem is disrupted. During the early stage of HF development (stage 6), the ring meristem begins to form as a ridge around the center of the flower. At this stage, the internal fourth-whorl organ is stem-like rather than carpelloid in MF. A total of 52,945 unigenes were identified as transcribed in MF and HF. A number of differentially expressed genes (DEGs) and metabolic pathways were detected as involved in the development of the gynoecium, especially the ovule, carpel and style. At the early gynoecium development stage, DEGs were shown to function in the metabolic pathways regulating ethylene biosynthesis and signal transduction (upstream regulator), auxin, cytokinin transport and signalling, and sex determination (or flower meristem identity). CONCLUSIONS: Pathways for the female sterility model were initially proposed to shed light on the molecular mechanisms of gynoecium development at early stages in T. sinensis.

Silk Fluorescence Collimator for Ultrasensitive Humidity Sensing and Light-Harvesting in Semitransparent Dye-Sensitized Solar Cells.

This work examines the self-collimation effect of silk materials on fluorescence emission/detection. A macroscopic regulation strategy, coupled with meso-reconstruction and meso-functionalization, is adopted to amplify the fluorescence emission of organic fluorescent dyes (i.e., Rhodamine 6G (R6G)) using silk photonic crystal (PC) films. The fluorescence emission can be linearly enhanced or inhibited by a PC as a result of the photonic bandgap coupling with the excitation light and/or emission light. Depending on the design of the silk fluorescence collimator, the emission can reach 49.37 times higher than the control. The silk fluorescence collimator can be applied to achieve significant benefits: for instance, as a humidity sensor, it provides good reproducibility and a sensitivity of 28.50 a.u./% relative humidity, which is 80.78 times higher than the sensitivity of the control, and as a novel curtain, it raises the energy conversion efficiency of the semitransparent dye-sensitized solar cells (DSSCs) by 16%.

The ethylene receptor regulates Typha angustifolia leaf aerenchyma morphogenesis and cell fate.

MAIN CONCLUSION: Ethylene receptor is crucial for PCD and aerenchyma formation in Typha angustifolia leaves. Not only does it receive and deliver the ethylene signal, but it probably can determine the cell fate during aerenchyma morphogenesis, which is due to the receptor expression quantity. Aquatic plant oxygen delivery relies on aerenchyma, which is formed by a programmed cell death (PCD) procedure. However, cells in the outer edge of the aerenchyma (palisade cells and septum cells) remain intact, and the mechanism is unclear. Here, we offer a hypothesis: cells that have a higher content of ethylene receptors do not undergo PCD. In this study, we investigated the leaf aerenchyma of the aquatic plant Typha angustifolia. Ethephon and pyrazinamide (PZA, an inhibitor of ACC oxidase) were used to confirm that ethylene is an essential hormone for PCD of leaf aerenchyma cells in T. angustifolia. That the ethylene receptor was an indispensable factor in this PCD was confirmed by 1-MCP (an inhibitor of the ethylene receptor) treatment. Although PCD can be avoided by blocking the ethylene receptor, excessive ethylene receptors also protect cells from PCD. TaETR1, TaETR2 and TaEIN4 in the T. angustifolia leaf were detected by immunofluorescence (IF) using polyclonal antibodies. The result showed that the content of ethylene receptors in PCD-unsusceptible cells was 4-14 times higher than that one in PCD-susceptible cells, suggesting that PCD-susceptible cells undergo the PCD programme, while PCD-unsusceptible cells do not due to the content difference in the ethylene receptor in different cells. A higher level of ethylene receptor content makes the cells insensitive to ethylene, thereby avoiding cell death and degradation.

Circularly Polarized States Spawning from Bound States in the Continuum.

Bound states in the continuum in periodic photonic systems like photonic crystal slabs are proved to be accompanied by vortex polarization singularities on the photonic bands in the momentum space. The winding structures of polarization states not only widen the field of topological physics but also show great potential that such systems could be applied in polarization manipulating. In this Letter, we report the phenomenon that by in-plane inversion (C_{2}) symmetry breaking, pairs of circularly polarized states could spawn from the eliminated bound states in the continuum. Along with the appearance of the circularly polarized states as the two poles of the Poincaré sphere together with linearly polarized states covering the equator, full coverage on the Poincaré sphere could be realized. As an application, ellipticity modulation of linear polarization is demonstrated in the visible frequency range. This phenomenon provides a new degree of freedom in modulating polarization. The C points could also find applications in light-matter interactions. Further studying and manipulating the reported polarization singularities may lead to novel phenomena and physics in radiation modulating and topological photonics.

Population genetics, phylogenomics and hybrid speciation of Juglans in China determined from whole chloroplast genomes, transcriptomes, and genotyping-by-sequencing (GBS).

Genomic data are a powerful tool for elucidating the processes involved in the evolution and divergence of species. The speciation and phylogenetic relationships among Chinese Juglans remain unclear. Here, we used results from phylogenomic and population genetic analyses, transcriptomics, Genotyping-By-Sequencing (GBS), and whole chloroplast genomes (Cp genome) data to infer processes of lineage formation among the five native Chinese species of the walnut genus (Juglans, Juglandaceae), a widespread, economically important group. We found that the processes of isolation generated diversity during glaciations, but that the recent range expansion of J. regia, probably from multiple refugia, led to hybrid formation both within and between sections of the genus. In southern China, human dispersal of J. regia brought it into contact with J. sigillata, which we determined to be an ecotype of J. regia that is now maintained as a landrace. In northern China, walnut hybridized with a distinct lineage of J. mandshurica to form J. hopeiensis, a controversial taxon (considered threatened) that our data indicate is a horticultural variety. Comparisons among whole chloroplast genomes and nuclear transcriptome analyses provided conflicting evidence for the timing of the divergence of Chinese Juglans taxa. J. cathayensis and J. mandshurica are poorly differentiated based our genomic data. Reconstruction of Juglans evolutionary history indicate that episodes of climatic variation over the past 4.5 to 33.80 million years, associated with glacial advances and retreats and population isolation, have shaped Chinese walnut demography and evolution, even in the presence of gene flow and introgression.

Observation of Polarization Vortices in Momentum Space.

The vortex, a fundamental topological excitation featuring the in-plane winding of a vector field, is important in various areas such as fluid dynamics, liquid crystals, and superconductors. Although commonly existing in nature, vortices were observed exclusively in real space. Here, we experimentally observed momentum-space vortices as the winding of far-field polarization vectors in the first Brillouin zone of periodic plasmonic structures. Using homemade polarization-resolved momentum-space imaging spectroscopy, we mapped out the dispersion, lifetime, and polarization of all radiative states at the visible wavelengths. The momentum-space vortices were experimentally identified by their winding patterns in the polarization-resolved isofrequency contours and their diverging radiative quality factors. Such polarization vortices can exist robustly on any periodic systems of vectorial fields, while they are not captured by the existing topological band theory developed for scalar fields. Our work provides a new way for designing high-Q plasmonic resonances, generating vector beams, and studying topological photonics in the momentum space.

Fimbrins 4 and 5 Act Synergistically During Polarized Pollen Tube Growth to Ensure Fertility in Arabidopsis.

The germination and polar growth of pollen are prerequisite for double fertilization in plants. The actin cytoskeleton and its binding proteins play pivotal roles in pollen germination and pollen tube growth. Two homologs of the actin-bundling protein fimbrin, AtFIM4 and AtFIM5, are highly expressed in pollen in Arabidopsis and can form distinct actin architectures in vitro, but how they co-operatively regulate pollen germination and pollen tube growth in vivo is largely unknown. In this study, we explored their functions during pollen germination and polar growth. Histochemical analysis demonstrated that AtFIM4 was expressed only after pollen grain hydration and, in the early stage of pollen tube growth, the expression level of AtFIM4 was low, indicating that it functions mainly during polarized tube growth, whereas AtFIM5 had high expression levels in both pollen grains and pollen tubes. Atfim4/atfim5 double mutant plants had fertility defects including reduced silique length and seed number, which were caused by severe defects in pollen germination and pollen tube growth. When the atfim4/atfim5 double mutant was complemented with the AtFIM5 protein, the polar growth of pollen tubes was fully rescued; however, AtFIM4 could only partially restore these defects. Fluorescence labeling showed that loss of function of both AtFIM4 and AtFIM5 decreased the extent of actin filament bundling throughout pollen tubes. Collectively, our results revealed that AtFIM4 acts co-ordinately with AtFIM5 to organize and maintain normal actin architecture in pollen grains and pollen tubes to fulfill double fertilization in plants.