Low temperature compensates for defective tapetum initiation to restore the fertility of the novel TGMS line ostms15.

In rice breeding, thermosensitive genic male sterility (TGMS) lines based on the tms5 locus have been extensively employed. Here, we reported a novel rice TGMS line ostms15 (Oryza sativa ssp. japonica ZH11) which show male sterility under high temperature and fertility under low temperature. Field evaluation from 2018 to 2021 revealed that its sterility under high temperature is more stable than that of tms5 (ZH11), even with occasional low temperature periods, indicating its considerable value for rice breeding. OsTMS15 encodes an LRR-RLK protein MULTIPLE SPOROCYTE1 (MSP1) which was reported to interact with its ligand to initiate tapetum development for pollen formation. In ostms15, a point mutation from GTA (Val) to GAA (Glu) in its TIR motif of the LRR region led to the TGMS phenotype. Cellular observation and gene expression analysis showed that the tapetum is still present in ostms15, while its function was substantially impaired under high temperature. However, its tapetum function was restored under low temperature. The interaction between mOsTMS15 and its ligand was reduced while this interaction was partially restored under low temperature. Slow development was reported to be a general mechanism of P/TGMS fertility restoration. We propose that the recovered protein interaction together with slow development under low temperature compensates for the defective tapetum initiation, which further restores ostms15 fertility. We used base editing to create a number of TGMS lines with different base substitutions based on the OsTMS15 locus. This work may also facilitate the mechanistic investigation and breeding of other crops.

Strategies and characterization methods for achieving high performance PEO-based solid-state lithium-ion batteries.

Polyethylene oxide (PEO) based polymer electrolytes have been widely used in solid-state lithium batteries (SSBs) owing to the high solubility of lithium salt, favourable ionic conductivity, flexibility for improved interfacial contact and scalable processing. In this work, we summarize the main limitations remaining to be solved before the large-scale commercialization of PEO-based SSBs, including (1) improving ionic conductivity toward high-rate performance and lower operating temperature, (2) enhancing mechanical strength for improved lithium dendrite resistance and large-scale processing, (3) strengthening electrochemical stability to match high energy density electrodes with high voltage, and (4) achieving high thermal stability toward safe operation. Meanwhile, the characterization methods to investigate the ion transportation mechanism, lithium dendrite growth and decomposition reaction are also discussed.

An Authentication and Secure Communication Scheme for In-Vehicle Networks Based on SOME/IP.

The rapid development of intelligent networked vehicles (ICVs) has brought many positive effects. Unfortunately, connecting to the outside exposes ICVs to security threats. Using secure protocols is an important approach to protect ICVs from hacker attacks and has become a hot research area for vehicle security. However, most of the previous studies were carried out on V2X networks, while those on in-vehicle networks (IVNs) did not involve Ethernet. To this end, oriented to the new IVNs based on Ethernet, we designed an efficient secure scheme, including an authentication scheme using the Scalable Service-Oriented Middleware over IP (SOME/IP) protocol and a secure communication scheme modifying the payload field of the original SOME/IP data frame. The security analysis shows that the designed authentication scheme can provide mutual identity authentication for communicating parties and ensure the confidentiality of the issued temporary session key; the designed authentication and secure communication scheme can resist the common malicious attacks conjointly. The performance experiments based on embedded devices show that the additional overhead introduced by the secure scheme is very limited. The secure scheme proposed in this article can promote the popularization of the SOME/IP protocol in IVNs and contribute to the secure communication of IVNs.

Delayed callose degradation restores the fertility of multiple P/TGMS lines in Arabidopsis.

Photoperiod/temperature-sensitive genic male sterility (P/TGMS) is widely applied for improving crop production. Previous investigations using the reversible male sterile (rvms) mutant showed that slow development is a general mechanism for restoring fertility to P/TGMS lines in Arabidopsis. In this work, we isolated a restorer of rvms-2 (res3), as the male sterility of rvms-2 was rescued by res3. Phenotype analysis and molecular cloning show that a point mutation in UPEX1 l in res3 leads to delayed secretion of callase A6 from the tapetum to the locule and tetrad callose wall degradation. Electrophoretic mobility shift assay and chromatin immunoprecipitation analysis demonstrated that the tapetal transcription factor ABORTED MICROSPORES directly regulates UPEX1 expression, revealing a pathway for tapetum secretory function. Early degradation of the callose wall in the transgenic line eliminated the fertility restoration effect of res3. The fertility of multiple known P/TGMS lines with pollen wall defects was also restored by res3. We propose that the remnant callose wall may broadly compensate for the pollen wall defects of P/TGMS lines by providing protection for pollen formation. A cellular mechanism is proposed to explain how slow development restores the fertility of P/TGMS lines in Arabidopsis.

Design of a CANFD to SOME/IP Gateway Considering Security for In-Vehicle Networks.

In recent years, Ethernet has been introduced into vehicular networks to cope with the increasing demand for bandwidth and complexity in communication networks. To exchange data between controller area network (CAN) and Ethernet, a gateway system is required to provide a communication interface. Additionally, the existence of networked devices exposes automobiles to cyber security threats. Against this background, a gateway for CAN/CAN with flexible data-rate (CANFD) to scalable service-oriented middleware over IP (SOME/IP) protocol conversion is designed, and security schemes are implemented in the routing process to provide integrity and confidentiality protections. Based on NXP-S32G, the designed gateway is implemented and evaluated. Under most operating conditions, the CPU and the RAM usage are less than 5% and 20 MB, respectively. Devices running a Linux operating system can easily bear such a system resource overhead. The latency caused by the security scheme accounts for about 25% of the entire protocol conversion latency. Considering the security protection provided by the security scheme, this overhead is worthwhile. The results show that the designed gateway can ensure a CAN/CANFD to SOME/IP protocol conversion with a low system resource overhead and a low latency while effectively resisting hacker attacks such as frame forgery, tampering, and sniffing.

Genome-wide Analysis of Histone Modifications Distribution using the Chromatin Immunoprecipitation Sequencing Method in Magnaporthe oryzae.

Chromatin immunoprecipitation sequencing (ChIP-seq) is a powerful and widely used molecular technique for mapping whole genome locations of transcription factors (TFs), chromatin regulators, and histone modifications, as well as detecting entire genomes for uncovering TF binding patterns and histone posttranslational modifications. Chromatin-modifying activities, such as histone methylation, are often recruited to specific gene regulatory sequences, causing localized changes in chromatin structures and resulting in specific transcriptional effects. The rice blast is a devastating fungal disease on rice throughout the world and is a model system for studying fungus-plant interaction. However, the molecular mechanisms in how the histone modifications regulate their virulence genes in Magnaporthe oryzae remain elusive. More researchers need to use ChIP-seq to study how histone epigenetic modification regulates their target genes. ChIP-seq is also widely used to study the interaction between protein and DNA in animals and plants, but it is less used in the field of plant pathology and has not been well developed. In this paper, we describe the experimental process and operation method of ChIP-seq to identify the genome-wide distribution of histone methylation (such as H3K4me3) that binds to the functional target genes in M. oryzae. Here, we present a protocol to analyze the genome-wide distribution of histone modifications, which can identify new target genes in the pathogenesis of M. oryzae and other filamentous fungi.

A parameter adaptive method for state of charge estimation of lithium-ion batteries with an improved extended Kalman filter.

An accurate state of charge (SOC) estimation in battery management systems (BMS) is of crucial importance to guarantee the safe and effective operation of automotive batteries. However, the BMS consistently suffers from inaccuracy of SOC estimation. Herein, we propose a SOC estimation approach with both high accuracy and robustness based on an improved extended Kalman filter (IEKF). An equivalent circuit model is established, and the simulated annealing-particle swarm optimization (SA-PSO) algorithm is used for offline parameter identification. Furthermore, improvements have been made with noise adaptation, a fading filter and a linear-nonlinear filtering based on the traditional EKF method, and rigorous mathematical proof has been carried out accordingly. To deal with model mismatch, online parameter identification is achieved by a dual Kalman filter. Finally, various experiments are performed to validate the proposed IEKF. Experimental results show that the IEKF algorithm can reduce the error to 2.94% under dynamic stress test conditions, and robustness analysis is verified with noise interference, hence demonstrating its practicability for extending to state estimation of battery packs applied in real-world operating conditions.

The COMPASS-like complex modulates fungal development and pathogenesis by regulating H3K4me3-mediated targeted gene expression in Magnaporthe oryzae.

Histone-3-lysine-4 (H3K4) methylation is catalysed by the multiprotein complex known as the Set1/COMPASS or MLL/COMPASS-like complex, an element that is highly evolutionarily conserved from yeast to humans. However, the components and mechanisms by which the COMPASS-like complex targets the H3K4 methylation of plant-pathogenic genes in fungi remain elusive. Here we present a comprehensive analysis combining biochemical, molecular, and genome-wide approaches to characterize the roles of the COMPASS-like family in the rice blast fungus Magnaporthe oryzae, a model plant pathogen. We purified and identified six conserved subunits of COMPASS from M. oryzae: MoBre2 (Cps60/ASH2L), MoSpp1 (Cps40/Cfp1), MoSwd2 (Cps35), MoSdc1 (Cps25/DPY30), MoSet1 (MLL/ALL), and MoRbBP5 (Cps50), using an affinity tag on MoBre2. We determined the sequence repeat in dual-specificity kinase splA and ryanodine receptors domain of MoBre2 can interact directly with the DPY30 domain of MoSdc1 in vitro. Furthermore, we found that deletion of the genes encoding COMPASS subunits of MoBre2, MoSPP1, and MoSwd2 caused similar defects regarding invasive hyphal development and pathogenicity. Genome-wide profiling of H3K4me3 revealed that it has remarkable co-occupancy at the transcription start site regions of target genes. Significantly, these target genes are often involved in spore germination and pathogenesis. Decreased gene expression caused by the deletion of MoBre2, MoSwd2, or MoSpp1 was highly correlated with a decrease in H3K4me3. These results suggest that MoBre2, MoSpp1, and MoSwd2 function as a whole COMPASS complex, contributing to fungal development and pathogenesis by regulating H3K4me3-targeted genes in M. oryzae.

OsMS188 Is a Key Regulator of Tapetum Development and Sporopollenin Synthesis in Rice.

BACKGROUND: During anther development, the tapetum provides essential nutrients and materials for pollen development. In rice, multiple transcription factors and enzymes essential for tapetum development and pollen wall formation have been cloned from male-sterile lines. RESULTS: In this study, we obtained several lines in which the MYB transcription factor OsMS188 was knocked out through the CRISPR-Cas9 approach. The osms188 lines exhibited a male-sterile phenotype with aberrant development and degeneration of tapetal cells, absence of the sexine layer and defective anther cuticles. CYP703A3, CYP704B2, OsPKS1, OsPKS2, DPW and ABCG15 are sporopollenin synthesis and transport-related genes in rice. Plants with mutations in these genes are male sterile, with a defective sexine layer and anther cuticle. Further biochemical assays demonstrated that OsMS188 binds directly to the promoters of these genes to regulate their expression. UDT1, OsTDF1, TDR, bHLH142 and EAT1 are upstream regulators of rice tapetum development. Electrophoretic mobility shift assays (EMSAs) and activation assays revealed that TDR directly regulates OsMS188 expression. Additionally, protein interaction assays indicated that TDR interacts with OsMS188 to regulate downstream gene expression. CONCLUSION: Overall, OsMS188 is a key regulator of tapetum development and pollen wall formation. The gene regulatory network established in this work may facilitate future investigations of fertility regulation in rice and in other crop species.

IMPERFECTIVE EXINE FORMATION (IEF) is required for exine formation and male fertility in Arabidopsis.

KEY MESSAGE: IEF, a novel plasma plasma membrane protein, is important for exine formation in Arabidopsis. Exine, an important part of pollen wall, is crucial for male fertility. The major component of exine is sporopollenin which are synthesized and secreted by tapetum. Although sporopollenin synthesis has been well studied, the transportation of it remains elusive. To understand it, we analyzed the gene expression pattern in tapetal microdissection data, and investigated the potential transporter genes that are putatively regulated by ABORTED MICROSPORES (AMS). Among these genes, we identified IMPERFECTIVE EXINE FORMATION (IEF) that is important for exine formation. Compared to the wild type, ief mutants exhibit severe male sterility and pollen abortion, suggesting IEF is crucial for pollen development and male fertility. Using both scanning and transmission electron microscopes, we showed that exine structure was not well defined in ief mutant. The transient expression of IEF-GFP driven by the 35S promoter indicated that IEF-GFP was localized in plasma membrane. Furthermore, AMS can specifically activate the expression of promoterIEF:LUC in vitro, which suggesting AMS regulates IEF for exine formation. The expression of ATP-BINDING CASSETTE TRANSPORTER G26 (AGCB26) was not affected in ief mutants. In addition, SEM and TEM data showed that the sporopollenin deposition is more defective in abcg26/ief-2 than that of in abcg26, which suggesting that IEF is involved in an independent sporopollenin transportation pathway. This work reveal a novel gene, IEF regulated by AMS that is essential for exine formation.

A Data-Driven Response Virtual Sensor Technique with Partial Vibration Measurements Using Convolutional Neural Network.

Measurement of dynamic responses plays an important role in structural health monitoring, damage detection and other fields of research. However, in aerospace engineering, the physical sensors are limited in the operational conditions of spacecraft, due to the severe environment in outer space. This paper proposes a virtual sensor model with partial vibration measurements using a convolutional neural network. The transmissibility function is employed as prior knowledge. A four-layer neural network with two convolutional layers, one fully connected layer, and an output layer is proposed as the predicting model. Numerical examples of two different structural dynamic systems demonstrate the performance of the proposed approach. The excellence of the novel technique is further indicated using a simply supported beam experiment comparing to a modal-model-based virtual sensor, which uses modal parameters, such as mode shapes, for estimating the responses of the faulty sensors. The results show that the presented data-driven response virtual sensor technique can predict structural response with high accuracy.

Magnetic Trapping of Cold Methyl Radicals.

We have demonstrated that a supersonic beam of methyl radicals (CH_{3}) in the ground rotational state of both para and ortho species has been slowed down to standstill with a magnetic molecular decelerator, and successfully captured spatially in an anti-Helmholtz magnetic trap for >1 s. The trapped CH_{3} radicals have a mean translational temperature of about 200 mK with an estimated density of >5.0×10^{7} cm^{-3}. The methyl radical is an ideal system for the study of cold molecules not only because of its high reactivities at low temperatures, but also because further cooling below 1 mK is plausible via sympathetic cooling with ultracold atoms. The demonstrated trapping capability of methyl radicals opens up various possibilities for realizing ultracold ensembles of molecules towards Bose-Einstein condensation of polyatomic molecules and investigations of reactions governed by quantum statistics.

Manipulation of translational motion of methyl radicals by pulsed magnetic fields.

Control of the translational motion of methyl radicals is demonstrated by using pulsed magnetic fields created by a series of solenoid coils. By synchronously switching the magnetic fields corresponding to the motion of the radicals, we have observed the deceleration and focusing of a supersonic radical beam. The present result shows that Zeeman deceleration of polyatomic free radicals in the doublet state can be realized. This opens possibilities for trapping cold polyatomic free radicals in a spatially confined area leading to further studies of cold collisions and reactions.

Scalable synthesis of highly reactive 1,3-diamino dienes from vinamidinium salts and their use in Diels-Alder reactions.

A practical and chromatography-free synthesis of vinamidinium salts and their use as diene precursors in Diels-Alder reactions is reported. Additionally, 1,3-dipyrrolidino-1,3-butadiene was shown to be significantly more reactive than Rawal's diene in a competition experiment.