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Abstract Background Cancer‐associated fibroblasts (CAFs) play a vital role in facilitating tumor progression through extensive reciprocal interplay with cancer cells. Tumor‐derived extracellular vesicles (EVs) are the critical mediators involved in the crosstalk between cancer cells and stromal cells, contributing to the metastasis of cancers. Yet, the biological mechanisms of tumor‐derived EVs in triggering CAFs phenotype to stimulate the lymph node (LN) metastasis of bladder cancer (BCa) are largely unknown. Here, we aimed to explore the effects and molecular mechanisms of tumor‐derived EV‐mediated CAFs phenotype in regulating BCa LN metastasis. Methods The high‐throughput sequencing was utilized to identify the crucial long non‐coding RNA (lncRNA) associated with CAF enrichment in BCa. The functional role of the transition of fibroblasts to CAFs induced by LINC00665‐mediated EVs was investigated through the in vitro and in vivo assays. Chromatin isolation by RNA purification assays, fluorescence resonance energy transfer assays, cytokine profiling and patient‐derived xenograft (PDX) model were performed to explore the underlying mechanism of LINC00665 in the LN metastasis of BCa. Results We found that CAFs are widely enriched in the tumor microenvironment of BCa, which correlated with BCa lymphangiogenesis and LN metastasis. We then identified a CAF‐associated long non‐coding RNA, LINC00665, which acted as a crucial mediator of CAF infiltration in BCa. Clinically, LINC00665 was associated with LN metastasis and poor prognosis in patients with BCa. Mechanistically, LINC00665 transcriptionally upregulated RAB27B expression and induced H3K4me3 modification on the promoter of RAB27B through the recruitment of hnRNPL. Moreover, RAB27B‐induced EVs secretion endowed fibroblasts with the CAF phenotype, which reciprocally induced LINC00665 overexpression to form a RAB27B‐HGF‐c‐Myc positive feedback loop, enhancing the lymphangiogenesis and LN metastasis of BCa. Importantly, we demonstrated that blocking EV‐transmitted LINC00665 or HGF broke this loop and impaired BCa lymphangiogenesis in a PDX model. Conclusion Our study uncovers a precise mechanism that LINC00665 sustains BCa LN metastasis by inducing a RAB27B‐HGF‐c‐Myc positive feedback loop between BCa cells and fibroblasts, suggesting that LINC00665 could be a promising therapeutic target for patients with LN metastatic BCa.

Abstract Lymph node (LN) metastasis is one of the predominant metastatic routes of non-small cell lung cancer (NSCLC) and is considered as a leading cause for the unsatisfactory prognosis of patients. Although lymphangiogenesis is well-recognized as a crucial process in mediating LN metastasis, the regulatory mechanism involving lymphangiogenesis and LN metastasis in NSCLC remains unclear. In this study, we employed high-throughput sequencing to identify a novel circular RNA (circRNA), circTLCD4-RWDD3, which was significantly upregulated in extracellular vesicles (EVs) from LN metastatic NSCLC and was positively associated with deteriorated OS and DFS of patients with NSCLC from multicenter clinical cohort. Downregulating the expression of EV-packaged circTLCD4-RWDD3 inhibited lymphangiogenesis and LN metastasis of NSCLC both in vitro and in vivo. Mechanically, circTLCD4-RWDD3 physically interacted with hnRNPA2B1 and mediated the SUMO2 modification at K108 residue of hnRNPA2B1 by upregulating UBC9. Subsequently, circTLCD4-RWDD3-induced SUMOylated hnRNPA2B1 was recognized by the SUMO interaction motif (SIM) of ALIX and activated ALIX to recruit ESCRT-III, thereby facilitating the sorting of circTLCD4-RWDD3 into NSCLC cell-derived EVs. Moreover, EV-packaged circTLCD4-RWDD3 was internalized by lymphatic endothelial cells to activate the transcription of PROX1, resulting in the lymphangiogenesis and LN metastasis of NSCLC. Importantly, blocking EV-mediated transmission of circTLCD4-RWDD3 via mutating SIM in ALIX or K108 residue of hnRNPA2B1 inhibited the lymphangiogenesis and LN metastasis of NSCLC in vivo. Our findings reveal a precise mechanism underlying SUMOylated hnRNPA2B1-induced EV packaging of circTLCD4-RWDD3 in facilitating LN metastasis of NSCLC, suggesting that EV-packaged circTLCD4-RWDD3 could be a potential therapeutic target against LN metastatic NSCLC.

Abstract Background Cancer-associated fibroblasts (CAFs) play pivotal roles in chemoresistance of pancreatic ductal adenocarcinoma (PDAC). However, the underlying mechanisms are poorly understood. Revealing the cross-talk network between tumor stroma and pancreatic cancer and developing effective strategies against oxaliplatin resistance are highly desired in the clinic. Methods High-throughput sequence was used to screened the key circRNAs transmitted by extracellular vesicles (EVs) from CAFs to pancreatic cancer cells. The associations between EV-packaged circBIRC6 and chemotherapy responsiveness were validated in a cohort of 82 cases of advanced PDAC patients. Then, the effects of EV-packaged circBIRC6 on CAF-induced oxaliplatin resistance were investigated by flow cytometry, colony formation, viability of pancreatic cancer organoids in vitro and by xenograft models in vivo. RNA pulldown, RNA immunoprecipitation, and sites mutation assays were used to reveal the underlying mechanism. Results We identified a circRNA, circBIRC6, is significantly upregulated in CAF-derived EVs and is positively associated with oxaliplatin-based chemoresistance. In vitro and in vivo functional assays showed that CAF-derived EV-packaged circBIRC6 enhance oxaliplatin resistance of pancreatic cancer cells and organoids via regulating the non-homologous end joining (NHEJ) dependent DNA repair. Mechanistically, circBIRC6 directly binds with XRCC4 and enhanced the interaction of XRCC4 with SUMO1 at the lysine 115 residue, which facilitated XRCC4 chromatin localization. XRCC4K115R mutation dramatically abrogated the EV-packaged circBIRC6 induced effect. Moreover, combination of antisense oligonucleotide inhibitors against circBIRC6 with Olaparib dramatically suppressed chemoresistance in patient-derived xenograft models. Conclusions Our study revealed that EV-packaged circBIRC6 confer oxaliplatin resistance in PDAC by mediating SUMOylation of XRCC4, introducing a promising predictive and therapeutic target for PDAC on oxaliplatin resistance.

Cucumber is one of the top ten vegetables globally and is widely cultivated worldwide. However, Fusarium wilt, caused by Fusarium oxysporum f. sp. Cucumerinum, is one of the most serious soil-borne diseases in cucumber cultivation, causing significant economic losses. Biological control has great potential in the prevention of cucumber wilt disease, but the mechanism involved still needs further research. In this study, biocontrol isolate Bacillus subtilis 1JN2, which was isolated in our previous work, was evaluated in field conditions against Fusarium wilt, and the rhizosphere fungal diversity was analyzed. The results indicated that the biocontrol efficacy of B. subtilis 1JN2 reached 58.5% compared with the blank control, and the population density of F. oxysporum in the rhizosphere decreased from 495 copies/g of soil before inoculation to 20 copies/g 14 days after treatment. High-throughput sequencing demonstrated that after an inoculation of 1JN2, the populations that decreased significantly include the genera of Olpidium and Pseudallescheria, from more than 20% to less than 8%. And the most increased population belonged to the family Chaetomiaceae, from 6.82% to 18.77%, 12.39%, 44.41%, and 19.41% at the four sample time points after treatment. In addition, soil-related enzyme activities, including catalase, soil dehydrogenase, alkaline phosphatase, and polyphenol oxidase, were analyzed before and after treatment with 1JN2. The results indicated that all the enzyme activities showed an upward trend following inoculation. These findings demonstrate the potential of using B. subtilis 1JN2 as a biocontrol agent for controlling Fusarium wilt in cucumber.

IntroductionAt present, the use of beneficial microorganisms to control cucumber Fusarium wilt is a widely used method, and the rhizosphere microecological reset is one of the mechanisms involved. However, how biocontrol strains reshape cucumber rhizosphere microecology remains to be further studied.MethodsThe composition changes of cucumber root exudates induced by biocontrol strain 1JN2, the microbial ecology of cucumber rhizosphere and the colonization ability of biocontrol strain 1JN2 in cucumber rhizosphere were analyzed through UHPLC-MS/MS analysis, Illumina high-throughput sequencing and SEM, respectively.ResultsFirst, cucumber plants treated with biocontrol Bacillus 1JN2 reduced the disease severity of Fusarium wilt by 60%. Significant changes in cucumber root exudates were found after 1JN2 inoculation and the contents of four amino acids including glutamine, tryptophan, glycine and glutamic acid were significantly increased. Second, It was found that the bacterial diversity in the rhizosphere of cucumber was significantly increased in both the strain treatment group and the amino acid mixture treatment group, The number of Bacillus was the largest in all dominant populations, exceeded 20% in all treatment groups. The bacteria of Hydrogenispora and Vicinamibacteria were significantly increased after treatment.DiscussionOverall, the results demonstrated that amino acid substances in cucumber root exudates induced by biocontrol strain 1JN2 can shift the cucumber root microenvironment and prevent the occurrence of Fusarium wilt disease.

Abstract Background Circular RNAs (circRNAs) circularized by back-splicing of pre-mRNA are widely expressed and affected the proliferation, invasion and metastasis of bladder cancer (BCa). However, the mechanism underlying circRNA biogenesis in mediating the distant metastasis of BCa still unexplored. Methods RNA sequencing data between BCa and normal adjacent tissues was applied to identify the differentially expressed circRNAs. The functions of circNIPBL in BCa were investigated via a series of biochemical experiments. The Clinical significance of circNIPBL was examined in a cohort of larger BCa tissues. Results In the present study, we identified a novel circRNA (hsa_circ_0001472), circNIPBL, which was significantly upregulated and had great influence on the poor prognosis of patients with BCa. Functionally, circNIPBL promotes BCa metastasis in vitro and in vivo. Mechanistically, circNIPBL upregulate the expression of Wnt5a and activated the Wnt/β-catenin signaling pathway via directly sponged miR-16-2-3p, leading to the upregulation of ZEB1, which triggers the EMT of BCa. Moreover, we revealed that ZEB1 interacted with the flanking introns of exons 2–9 on NIPBL pre-mRNA to trigger circNIPBL biogenesis, thus forming a positive feedback loop. Importantly, circNIPBL overexpression significantly facilitated the distant metastasis of BCa in the orthotopic bladder cancer model, while silencing ZEB1 remarkably blocked the effects of metastasis induced by circNIPBL overexpression. Conclusions Our study highlights that circNIPBL-induced Wnt signaling pathway activation triggers ZEB1-mediated circNIPBL biogenesis, which forms a positive feedback loop via the circNIPBL/miR-16-2-3p/Wnt5a/ZEB1 axis, supporting circNIPBL as a novel therapeutic target and potential biomarker for BCa patients. Graphical Abstract

Today, aviation has grown significantly in importance. However, the challenge of flight delays has become increasingly severe due to the need for safe separation between aircraft to mitigate wake turbulence effects. The primary emphasis of this investigation resides in elucidating the evolutionary attributes of wake vortices in homogeneous isotropy turbulence. The large eddy simulation (LES) method is used to scrutinize the dynamic evolution of wake vortices engendered by an A333 aircraft in the atmospheric milieu and assess its ramifications on the ARJ21 aircraft. The research endeavor commences by formulating an LES methodology for the evolution of aircraft wake vortices, integrating adaptive grid technology to reduce the necessary grid volume significantly. This approach enables the implementation of axial and non-axial grid adaptive refinement, leading to more accurate simulations of both axial and non-axial vortices. Numerical simulations are conducted using the LES approach to scrutinize three distinct rates of turbulence dissipation amidst the ambient atmospheric turbulence, and the results are juxtaposed with Lidar measurements (Wind3D 6000 LiDAR) of wake vortices acquired at Chengdu Shuangliu International Airport (CTU). Subsequently, the rolling moment of the following aircraft is calculated, and three-dimensional hazard zones are determined for the A333. It is found that during the approach phase, the wake turbulence separation minima for an ARJ21 (CAT-F) following an A333 (CAT-B) is 3.35 NM, which represents a reduction of approximately 33% compared to ICAO RECAT (Wake Turbulence Re-categorization). The findings validate the dependability of the fine-grained mesh used in the vortex core region, engendered through the adaptive grid method, which proficiently captures the Crow instability and the interconnected phenomena of vortices in the numerical examination of aircraft wake. The safety of wake encounters primarily depends on the magnitude of environmental turbulence and the development of structural instability in wake vortices.

Genome sequencing is widely recognized as a fundamental pillar in genetic research and legal studies of biological phenomena, providing essential insights for genetic investigations and legal analyses of biological events. The field of genome sequencing has experienced significant progress due to rapid improvements in scientific and technological developments. These advancements encompass not only significant improvements in the speed and quality of sequencing but also provide an unparalleled opportunity to explore the subtle complexities of genomes, particularly in the context of rare species. Such a wide range of possibilities has successfully supported the validation of plant gene functions and the refinement of precision breeding methodologies. This expanded scope now includes a comprehensive exploration of the current state and conservation efforts of gymnosperm gene sequencing, offering invaluable insights into their genomic landscapes. This comprehensive review elucidates the trajectory of development and the diverse applications of genome sequencing. It encompasses various domains, including crop breeding, responses to abiotic stress, species evolutionary dynamics, biodiversity, and the unique challenges faced in the conservation and utilization of gymnosperms. It highlights both ongoing challenges and the unveiling of forthcoming developmental trajectories.

【Objective】The study was carried out to explore the effect of chemical stress on the colonization ability of biocontrol bacteria on cucumber plants.【Method】By setting a greenhouse experiment and two field experiments, biocontrol bacteria were inoculated on cucumber plants without fungicides and pathogens, with fungicides and without pathogens, with fungicides and pathogens, respectively.【Result】The six biocontrol strains tested had good colonization effects on cucumber leaves and roots. Under the greenhouse conditions without fungicides and pathogens, the colonization of each biocontrol strain showed a trend of decrease-increase-stability, strains 1BS4 and 3BY4 had strong colonization ability in cucumber leaves and roots. The colonization quantity of strains 1BS4 and 3BY4 was close to 106 CFU/g(FW) at 30 days post inoculation(dpi) about 104 CFU/g(FW); Under the field conditions without fungicides and with pathogens, there were significant differences in the colonization quantities of various biocontrol strains in leaves and roots, and they all showed a trend of decrease-increase-decrease-stability. Strain 5BJN1 had the strongest colonization ability. The colonization quantity of cucumber leaves and roots at 30 dpi was 105.92 CFU/g(FW) and 104.95 CFU/g(FW), respectively; Under the field conditions with both fungicides and pathogens, there were differences in the colonization quantity of biocontrol bacteria in leaves and roots, but the difference was not significant, and all showed a trend of decrease-increase-decrease-stability, and the colonization quantities of strains 3BJN7 and 5BJN1 in cucumber leaves at 30 dpi were relatively high, which were 103.88 CFU/g(FW) and 104.23 CFU/g(FW), respectively. The colonization quantity of other strains was about 103 CFU/g(FW).【Conclusion】The effect of chemicals on the colonization ability of biocontrol bacteria depends on the types of strains. Some strains (3BJN7 and 5BJN1) have good compatibility with chemicals. In the presence of cucumber downy mildew, they can still maintain a certain amount of colonization after using chemicals, therefore, they can be used as potential biocontrol agents; In the absence of fungicide stress, the emergence of downy mildew pathogen can induce the improvement of colonization ability of strain 3BJN7 so that it can colonize stably on cucumber plants.

The TEOSINTE BRANCHED1 (TBI1), CYCLOIDEA (CYC), and PROLIFERATING CELL NUCLEAR ANTIGEN FACTORS (PCF1 and PCF2) proteins truncated as TCP transcription factors carry conserved basic-helix-loop-helix (bHLH) structure, related to DNA binding functions. Evolutionary history of the TCP genes has shown their presence in early land plants. In this paper, we performed a comparative discussion on the current knowledge of the TCP Transcription Factors in lower and higher plants: their evolutionary history based on the phylogenetics of 849 TCP proteins from 37 plant species, duplication events, and biochemical roles in some of the plants species. Phylogenetics investigations confirmed the classification of TCP TFs into Class I (the PCF1/2), and Class II (the C- clade) factors; the Class II factors were further divided into the CIN- and CYC/TB1- subclade. A trace in the evolution of the TCP Factors revealed an absence of the CYC/TB1subclade in lower plants, and an independent evolution of the CYC/TB1subclade in both eudicot and monocot species. 54% of the total duplication events analyzed were biased towards the dispersed duplication, and we concluded that dispersed duplication events contributed to the expansion of the TCP gene family. Analysis in the TCP factors functional roles confirmed their involvement in various biochemical processes which mainly included promoting cell proliferation in leaves in Class I TCPs, and cell division during plant development in Class II TCP Factors. Apart from growth and development, the TCP Factors were also shown to regulate hormonal and stress response pathways. Although this paper does not exhaust the present knowledge of the TCP Transcription Factors, it provides a base for further exploration of the gene family.

Utilization of rice blast-resistance (R) genes is the most economical and environmentally friendly method to control blast disease. However, rice varieties with R genes influence the outcome of genetic architectures of Magnaporthe oryzae (M. oryzae), and mutations in avirulence (AVR) genes of M. oryzae may cause dysfunction of the corresponding R genes in rice varieties. Although monitoring and characterizing rice R genes and pathogen AVR genes in field populations may facilitate the implementation of effective R genes, little is known about the changes of R genes over time and their ultimate impact on pathogen AVR genes. In this study, 117 main cultivated rice varieties over the past five decades and 35 M. oryzae isolates collected from those diseased plants were analyzed by PCR using gene-specific markers of the nine R genes and six primer pairs targeting the coding sequence or promoter of AVR genes, respectively. The R genes Pigm, Pi9, Pi2, Piz-t, Pi-ta, Pik, Pi1, Pikp, and Pikm were identified in 5, 0, 1, 4, 18, 0, 2, 1, and 0 cultivars, respectively. Significantly, none of these R genes had significant changes that correlated to their application periods of time. Among the four identified AVR genes, AVR-Pik had the highest amplification frequency (97.14%) followed by AVR-Pita (51.43%) and AVR-Pi9 (48.57%); AVR-Piz-t had the lowest frequency (28.57%). All these AVR genes except AVR-Pi9 had 1–2 variants. Inoculation mono-genic lines contained functional genes of Pi2/9 and Pik loci with 14 representative isolates from those 35 ones revealed that the presence of certain AVR-Piz-t, AVR-Pita variants, and AVR-Pik-E + AVR-Pik-D in M. oryzae populations, and these variants negated the ability of the corresponding R genes to confer resistance. Importantly, Pi2, Pi9, and Pigm conferred broad-spectrum resistance to these local isolates. These findings reveal that the complex genetic basis of M. oryzae and some effective blast R genes should be considered in future rice blast-resistance breeding programs.

This study examines the mobile phone practices of rural left-behind children (LBC) whose one or both parents migrate to cities for better earnings and the impact of such practices on migrant families in China. The study has used ethnographic approach by conducting participant observations and interviews of 21 LBC, residing in Guangren village, south China's Guangxi Autonomous Region. The study uses domestication theory to analyze these LBC's adoption of mobile phones in their daily routines and spaces in and out of their households. The key findings are as follows: (a) the LBC used mobile phones primarily to engage with their distant parent(s); (b) through collaborative efforts, they tried to enhance familial connections; and (c) they overcome the separation issue by co-participating in ongoing events, thus making the domestication of mobile phone a distant solving of real-world problems faced by migrant parent(s) and their LBC. The study concludes that LBC's innovative uses of mobile phones empowered them by building shared virtual space with their migrant parent(s), via which they handled the separation issue. In such shared virtual spaces, LBC's families have developed rich expressions of familial connections in various forms based on the limited perpetuate connectedness.

Biofilm plays important roles in the life cycle of Bacillus species, such as promoting host and object surface colonization and resisting heavy metal stress. This study utilized transcriptomics to evaluate the impacts of cadmium on the components, morphology, and function of biofilms of Bacillus subtilis strain 1JN2. Under cadmium ion stress, the morphology of the B. subtilis 1JN2 biofilm was flattened, and its mobility increased. Moreover, differential gene expression analysis showed that the main regulator of biofilm formation, Spo0A, decreased in expression under cadmium ion stress, thereby inhibiting extracellular polysaccharide synthesis through the SinI/SinR two-component regulatory system and the AbrB pathway. Cadmium ion treatment also increased the SigD content significantly, thereby increasing the expression of the flagella encoding and assembly genes in the strain. This promoted poly-γ-glutamic acid production via the DegS/DegU two-component regulatory system and the conversion of biofilm extracellular polysaccharide to poly-γ-glutamic acid. This conferred cadmium stress tolerance in the strain. Additionally, the cadmium ion-mediated changes in the biofilm composition affected the colonization of the strain on the host plant root surface. Cadmium ions also induced surfactin synthesis. These findings illustrate the potential of Bacillus species as biocontrol strains that can mitigate plant pathogenic infections and heavy metal stress. The results also provide a basis for the screening of multifunctional biocontrol strains.

Lymph node (LN) metastasis occurs frequently in pancreatic ductal adenocarcinoma (PDAC) and predicts poor prognosis for patients. The KRASG12D mutation confers an aggressive PDAC phenotype that is susceptible to lymphatic dissemination. However, the regulatory mechanism underlying KRASG12D mutation–driven LN metastasis in PDAC remains unclear. Herein, we found that PDAC with the KRASG12D mutation (KRASG12D PDAC) sustained extracellular vesicle–mediated (EV-mediated) transmission of heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1) in a SUMOylation-dependent manner and promoted lymphangiogenesis and LN metastasis in vitro and in vivo. Mechanistically, hnRNPA1 bound with SUMO2 at the lysine 113 residue via KRASG12D-induced hyperactivation of SUMOylation, which enabled its interaction with TSG101 to enhance hnRNPA1 packaging and transmission via EVs. Subsequently, SUMOylation induced EV-packaged-hnRNPA1 anchoring to the adenylate- and uridylate-rich elements of PROX1 in lymphatic endothelial cells, thus stabilizing PROX1 mRNA. Importantly, impeding SUMOylation of EV-packaged hnRNPA1 dramatically inhibited LN metastasis of KRASG12D PDAC in a genetically engineered KrasG12D/+ Trp53R172H/+ Pdx-1-Cre (KPC) mouse model. Our findings highlight the mechanism by which KRAS mutant–driven SUMOylation triggers EV-packaged hnRNPA1 transmission to promote lymphangiogenesis and LN metastasis, shedding light on the potential application of hnRNPA1 as a therapeutic target in patients with KRASG12D PDAC.

Aircraft wake is a kind of intense air movement, and the study of its generation, development, and dissipation law is of great significance to the flight safety. There are abundant researches on the evolution of aircraft wakes affected by weather and ground effects; however, there are few studies on the influence of a single obstacle on the evolution of aircraft wake. In this article, in order to explore the influence of a single obstacle on the evolution of aircraft wake, firstly, we develop a computational fluid dynamics-based method of simulation of aircraft wake affected by cubic obstacle of different heights in order to obtain the wake intensity changes and position changes before and after being affected by the obstacle. Then, the result data are visualized and analyzed, and we obtain the results of velocity and Q criterion contours, circulation, and data related to wake vortex structure. CFD simulations are conducted, including the cases of the vertical distance between wake vortex and obstacle which is 20 m, 60 m, 100 m, and no obstacle. The quantitative results indicate that a single obstacle also has a great influence on the evolution of the wake vortex. Obstacle will shorten the time for the wake vortex to enter the fast decay stage, and the smaller the distance the wake vortex is above the obstacle, the faster it enters the fast decay stage. In the same time, the circulation will reduce 20% more under the same calculation time when the wake is 20 m above the obstacle than when the wake is 100 m above the obstacle, and the circulation will reduce 45% more than when there is no obstacle. Single obstacle also leads to the generation of multiple secondary vortices and rotates around the wake vortex, resulting in the increase of wake vortex core radius, wake vortex core spacing, and wake vortex height.

In order to shorten the wake safe separation, numerical simulation technology with aerodynamic response models and strip models have been combined to calculate wake hazard zone. As a realistic case, a medium aircraft ARJ21 following a heavy aircraft A330-200 is considered, and the Reynolds-averaged Navier-Stokes (RANS) method is used to explore the wake vortex evolution process of the leading aircraft at the decision height. A strip algorithm is proposed to calculate the rolling moment coefficient and overload increment of the ARJ21 after encountering the wake of the front aircraft in the three-dimensional space. The proposed algorithm identifies the area where the wake of the front aircraft can cause risks to the following aircraft and analyzes the evolution process of the hazard zone of the section where the decision height is located. The minimum safe separation of the ARJ21 following the A330-200 is 1.32 nmile, which is 26.4% of the ICAO separation standard of 5 nmile. When the average runway occupancy time (ROT) is reduced to match the separation of 1.32 nmile, the capacity of runway 02R/20L in Tianfu International Airport could theoretically reach 102.37 sorties/h under this aircraft pair combination. Compared to original 27 sorties/h, the runway capacity can be improved up to 279.14%, which will increase the airport operation efficiency.

To ensure the safety of aircraft operation, the current regional passenger aircraft maintains a large distance from the preceding aircraft in actual operation, which result in reducing the operation efficiency of airports and airspace, and increasing pollutant emissions. To address these issues, in this paper, two aircraft types are selected in which the CRJ-900 encounters the trailing wake vortices of the A380 in front. An improved strip model is developed to build the CRJ-900 overall response wake encounter value. First, the safety of the CRJ-900 longitudinal and lateral wake encounters in different flight stages is analyzed. Second, we calculate the critical safety separation and its impact on air transport efficiency. Third, we use the LTO model to measure the reduction of aircraft fuel consumption and pollutant emissions. The results demonstrated that the medium-sized aircraft CRJ-900 has the potential to reduce the wake separation when following the super-heavy A380 aircraft. In terms of the critical safety separation calculated by the safety index, the operating efficiency of airports and airspace could be effectively improved, allowing the reduction of pollutant emissions during aircraft take-off and landing. During the takeoff, level flight, and landing phase, the results are summarized as follows: when the CRJ-900 is 13km away from the A380, the maximum lift variation is 11334N, 8157N, and 7366N; the maximum rolling moment variation is 43836N•M, 35274 N•M, and 28487 N•M; the maximum value of the rolling moment coefficient (RMC) is 0.0171, 0.0160, and 0.0130; when the RMC critical value is 0.031, the maximum safe separation for different flight stages is 11960m, which is 1040m shorter than the existing separation; when the RMC critical value is 0.05, the maximum safe separation distance of each stage is 10083m, a reduction of 2917m compared with the existing separation; when the RMC threshold is 0.07, the maximum safe separation of different flight stages is 9021m, a reduction of 3979m compared to the existing separation; when the RMC value is between 0.031-0.07, the fuel consumption can be reduced by 7.9%–12.8%, and the pollutant emission can be reduced by 9.1%–12.8%.

Traditional research on static wake-vortex reduction usually considers only the influence of external environmental factors, while ignoring the dynamic change in an aircraft’s flight state. In order to solve this problem, this paper proposes a method to reduce separation using dynamic wake information based on changes in flight velocity. Firstly, relying on the wake-vortex generation and dissipation model, the initial circulations and dissipation parameters of the wake vortex at different aircraft velocities are calculated. Then, the complete evolution process of the wake vortex generated by different types of aircraft at different velocities is analyzed, and the evolution law of a wake vortex with changing velocity is obtained. Afterwards, according to the actual aerodynamic shape of CRJ-900, the aerodynamic model for CRJ-900 when it encounters a wake vortex is established. The situation of an CRJ-900 encountering a wake vortex under TBS is analyzed, which proves that TBS is safe and has a certain reduction potential. Then, taking the rolling moment coefficient as the safety index, the minimum safe separations at different velocities are calculated. Finally, a simulation for the separation reduction based on a dynamic wake vortex is carried out. Compared with the static wake-based separation, the dynamic wake-based separation technology can greatly reduce the aircraft separation requirements while ensuring the operation safety. The final simulation results show that the dynamic separation reduction in CRJ-900 following a medium and heavy aircraft can reach 44.3% and 51.6%, respectively.

Plant defense and growth rely on multiple transcriptional factors (TFs). Repression of shoot growth (RSG) is a TF belonging to a bZIP family in tobacco, known to be involved in plant gibberellin feedback regulation by inducing the expression of key genes. The tobacco calcium-dependent protein kinase CDPK1 was reported to interact with RSG and manipulate its intracellular localization by phosphorylating Ser-114 of RSG previously. Here, we identified tobacco mitogen-activated protein kinase 3 (NtMPK3) as an RSG-interacting protein kinase. Moreover, the mutation of the predicted MAPK-associated phosphorylation site of RSG (Thr-30, Ser-74, and Thr-135) significantly altered the intracellular localization of the NtMPK3-RSG interaction complex. Nuclear transport of RSG and its amino acid mutants (T30A and S74A) were observed after being treated with plant defense elicitor peptide flg22 within 5 min, and the two mutated RSG swiftly re-localized in tobacco cytoplasm within 30 min. In addition, triple-point mutation of RSG (T30A/S74A/T135A) mimics constant unphosphorylated status, and is predominantly localized in tobacco cytoplasm. RSG (T30A/S74A/T135A) showed no re-localization effect under the treatments of flg22, B. cereus AR156, or GA3, and over-expression of this mutant in tobacco resulted in lower expression levels of downstream gene GA20ox1. Our results suggest that MAPK-associated phosphorylation sites of RSG regulate its localization in tobacco, and that constant unphosphorylation of RSG in Thr-30, Ser-74, and Thr-135 keeps RSG predominantly localized in cytoplasm.

Rice blast disease caused by a fungus, Magnaporthe grisea, is one of the most destructive diseases in rice production worldwide, and salicylic acid (SA) can efficiently decrease the damage of M. grisea. Here, we combined the 2-Dimensional-Liquid Chromatography and the Matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (2D-LC-MALDI-TOF-TOF MS) techniques to compare and identify differentially expressed labelled proteins by the isobaric tags for relative and absolute quantitation (iTRAQ) between the blast-resistant cultivar Minghui and the susceptible rice cultivar Nipponbare in response to blast fungus infection. The group samples were treated with salicylic acid and compared to control samples. A total of 139 DEPs from the two cultivars showed either more than a two-fold change or alternating regulation patterns. Protein functionality analysis also exhibited that these proteins are involved in a wide range of molecular functions including: energy-related activity (30%), signal transduction (11%), redox homeostasis (15%), amino acid and nitrogen metabolism (4%), carbohydrate metabolism (5%), protein folding and assembly (10%), protein hydrolysis (9%), protein synthesis (12%), and other unknown functions (4%). Specifically, we demonstrated that exogenous treatment with salicylic acid promoted recovery in both rice cultivars from Magnaporthe grisea infection by enhancing: the regulation of signal transduction, increasing energy conversion and production through the regulation of the glycolytic pathway, and other various biochemical processes. These findings may facilitate future studies of the molecular mechanisms of rice blast resistance.

Carbon monoxide (CO) and nitric oxide (NO) are signal molecules that enhance plant adaptation to environmental stimuli. Auxin is an essential phytohormone for plant growth and development. CO and NO play crucial roles in modulating the plant’s response to iron deficiency. Iron deficiency leads to an increase in the activity of heme oxygenase (HO) and the subsequent generation of CO. Additionally, it alters the polar subcellular distribution of Pin-Formed 1 (PIN1) proteins, resulting in enhanced auxin transport. This alteration, in turn, leads to an increase in NO accumulation. Furthermore, iron deficiency enhances the activity of ferric chelate reductase (FCR), as well as the expression of the Fer-like iron deficiency-induced transcription factor 1 (FIT) and the ferric reduction oxidase 2 (FRO2) genes in plant roots. Overexpression of the long hypocotyl 1 (HY1) gene, which encodes heme oxygenase, or the CO donor treatment resulted in enhanced basipetal auxin transport, higher FCR activity, and the expression of FIT and FRO2 genes under Fe deficiency. Here, a potential mechanism is proposed: CO and NO interact with auxin to address iron deficiency stress. CO alters auxin transport, enhancing its accumulation in roots and up-regulating key iron-related genes like FRO2 and IRT1. Elevated auxin levels affect NO signaling, leading to greater sensitivity in root development. This interplay promotes FCR activity, which is crucial for iron absorption. Together, these molecules enhance iron uptake and root growth, revealing a novel aspect of plant physiology in adapting to environmental stress. [ABSTRACT FROM AUTHOR]

Biofilm plays important roles in the life cycle of Bacillus species, such as promoting host and object surface colonization and resisting heavy metal stress. This study utilized transcriptomics to evaluate the impacts of cadmium on the components, morphology, and function of biofilms of Bacillus subtilis strain 1JN2. Under cadmium ion stress, the morphology of the B. subtilis 1JN2 biofilm was flattened, and its mobility increased. Moreover, differential gene expression analysis showed that the main regulator of biofilm formation, Spo0A, decreased in expression under cadmium ion stress, thereby inhibiting extracellular polysaccharide synthesis through the SinI/SinR two-component regulatory system and the AbrB pathway. Cadmium ion treatment also increased the SigD content significantly, thereby increasing the expression of the flagella encoding and assembly genes in the strain. This promoted poly-γ-glutamic acid production via the DegS/DegU two-component regulatory system and the conversion of biofilm extracellular polysaccharide to poly-γ-glutamic acid. This conferred cadmium stress tolerance in the strain. Additionally, the cadmium ion-mediated changes in the biofilm composition affected the colonization of the strain on the host plant root surface. Cadmium ions also induced surfactin synthesis. These findings illustrate the potential of Bacillus species as biocontrol strains that can mitigate plant pathogenic infections and heavy metal stress. The results also provide a basis for the screening of multifunctional biocontrol strains. [ABSTRACT FROM AUTHOR]