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Image-Based Overlay (IBO) equipment leverages optical reflection imaging principles, combined with focusing and alignment strategies to measure overlay marks. Among all measurement steps, the focal plane measurement of marks exerts the most critical impact on overlay accuracy, while the time consumed by focal plane detection directly determines the overall measurement throughput. To address the trade-off between accuracy and efficiency in advanced process nodes, this paper proposes an integrated optimization strategy encompassing optical hardware design and software algorithms. The hardware solution adopts a dual-wavelength, dual-detector architecture: optimal imaging wavelengths are selected independently for the previous-layer and current-layer marks, ensuring each layer achieves ideal imaging conditions without mutual interference. The software strategy employs a deep learning framework to simultaneously predict and adjust the horizontal position (alignment) and vertical defocus number of measured marks in real time with high precision, thereby securing the optimal imaging posture. By synergizing hardware-based optimal imaging conditions and software-based posture adjustment, this method effectively mitigates the impact of background noise and system aberrations, ultimately improving both the accuracy and efficiency of overlay measurement.

Background Chinese chestnut ( Castanea mollissima ) is an economically and ecologically important woody nut crop. In C. mollissima , flowering is fundamental for nut yield. The MADS-box gene APETALA 1 ( AP1 ) plays essential roles in floral initiation and floral organ development in many plants. However, the function of the AP1 gene CmAP1 in C. mollissima is still unclear. Here, we cloned the coding sequence (CDS) and promoter of CmAP1 and analyzed the function of this gene. Results The CDS of CmAP1 is 741 bp and encodes a 246–amino acid protein. Subcellular analysis revealed that CmAP1 localizes to the nucleus. GUS driven by the CmAP1 promoter was expressed in seedlings and in leaf margins, petals, and carpels of transgenic Arabidopsis ( Arabidopsis thaliana ). RNA in situ hybridization indicated that CmAP1 was mainly expressed in the inflorescence meristem, floral primordia, sepal primordia, petal primordia, stamen primordia, and carpel primordia during the early stage of flower development. An auxin response element (TGA element), jasmonic acid response element (TGACG motif), and WRKY binding site (W-box element) were identified in the CmAP1 promoter. Heterologous expression of CmAP1 in wild-type and ap1-11 Arabidopsis resulted in early flowering. Notably, the expression of CmAP1 rescued the loss of the petal whorl in the ap1-11 mutant. AtAP1 , AtSEP1 , AtSEP2 , AtSEP3 , and AtSEP4 were upregulated in CmAP1 -expressing Arabidopsis plants. Conclusions These findings suggest that CmAP1 promotes flowering and plays a key role in petal development. Our findings help reveal the regulatory mechanism of flowering and flower development in C. mollissima , providing a practical basis for increasing yield in Chinese chestnut.

Naringenin is an essential precursor for all flavonoids, and effectively promoting naringenin production is crucial in metabolic engineering. The interaction between plant metabolic enzymes ensures metabolic flux. The effect can effectively improve the natural product synthesis of engineering microbial systems. In this study, chalcone isomerase genes in Allium fistulosum have been identified. The expression of AfCHIL is closely related to the accumulation of anthocyanins, and the expression of AfCHIL and AfCHS was highly synchronized. Yeast two-hybrid and firefly luciferase complementation imaging assay further confirmed AfCHIL physically interacted with AfCHS/AfCHI. The bioconversion experiment confirmed that AfCHIL reduced the derailment produced by AfCHS and increased the yield of naringenin. In addition, a system of biosynthesis naringenin involved in AfCHS was constructed, and these results suggested that the potential function between CHS with CHIL advanced naringenin production effectively. In conclusion, this study illustrated the function of AfCHIs in Allium fistulosum and provided new insight into improving the synthesis efficiency of naringenin.

Chinese chestnut ( Castanea mollissima Blume) is one of the earliest domesticated plants of high nutritional and ecological value, yet mechanisms of C. mollissima underlying its growth and development are poorly understood. Although individual chestnut species differ greatly, the molecular basis of the formation of their characteristic traits remains unknown. Though the draft genomes of chestnut have been previously released, the pan-genome of different variety needs to be studied. We report the genome sequence of three cultivated varieties of chestnut herein, namely Hei-Shan-Zhai-7 (H7, drought-resistant variety), Yan-Hong (YH, easy-pruning variety), and Yan-Shan-Zao-Sheng (ZS, early-maturing variety), to expedite convenience and efficiency in its genetics-based breeding. We obtained three chromosome-level chestnut genome assemblies through a combination of Oxford Nanopore technology, Illumina HiSeq X, and Hi-C mapping. The final genome assemblies are 671.99 Mb (YH), 790.99 Mb (ZS), and 678.90 Mb (H7), across 12 chromosomes, with scaffold N50 sizes of 50.50 Mb (YH), 65.05 Mb (ZS), and 52.16 Mb (H7). Through the identification of homologous genes and the cluster analysis of gene families, we found that H7, YH and ZS had 159, 131, and 91 unique gene families, respectively, and there were 13,248 single-copy direct homologous genes in the three chestnut varieties. For the convenience of research, the chestnut genome database 1 was constructed. Based on the results of gene family identification, the presence/absence variations (PAVs) information of the three sample genes was calculated, and a total of 2,364, 2,232, and 1,475 unique genes were identified in H7, YH and ZS, respectively. Our results suggest that the GBSS II-b gene family underwent expansion in chestnut (relative to nearest source species). Overall, we developed high-quality and well-annotated genome sequences of three C. mollissima varieties, which will facilitate clarifying the molecular mechanisms underlying important traits, and shortening the breeding process.

The preservation of biodiversity is crucial for maintaining ecological balance and promoting the harmonious development of mankind and nature. To formulate a biodiversity conservation plan for Guilin in China and accurately evaluate the impact of conservation measures on regional biodiversity, this study combined the InVEST model (v 3.7.0), the GeoSOS-FLUS model (v 2.3), and the landscape pattern index, analyzing the spatiotemporal changes in biodiversity from 2000 to 2020 in Guilin and simulating biodiversity under different development scenarios in 2040. The results showed the following: (1) The biodiversity index in Guilin displayed a declining trend, with the average annual value decreasing from 0.875 in 2000 to 0.870 in 2020. The area experiencing a reduction in biodiversity was primarily concentrated around the city. (2) The biodiversity level of each district and county had a large spatiotemporal difference, with the overall distribution characteristics of “higher in the northwest, southwest, and east, and lower in the northeast, southeast and central”. (3) The biodiversity hotspots were identified as priority areas for conservation; woodland and wetland were effectively protected, and the expansion of construction land and arable land was limited under the ecological protection scenario compared with the natural development scenario. (4) The annual average value of the biodiversity index of Guilin could reach 0.872 in 2040 after the adoption of ecological conservation measures, which is 0.013 higher than the natural development scenario. The results of this study can provide guidance for the construction of a national sustainable development model city and land use planning in Guilin, as well as a scientific reference for the creation of biodiversity conservation policies.

Chestnut (Castanea mollissima) is an economically important forest tree species, and its flowers possess functions such as repelling mosquitoes, killing bacteria, and clearing heat. However, the regulatory mechanisms of floral volatile organic compounds (VOCs) in chestnut are still unclear. This study analyzed the contents of major volatile compounds and related gene expression levels in chestnut flowers during the initial flowering stage (IFS) and full-flowering stage (FFS) using metabolomics and transcription techniques. In total, 926 volatile compounds were detected, mainly terpenes, heterocyclic compounds, and esters. Acetylenone, styrene, and β-pinene had contents that exceeded 5% in FFS chestnut flowers. In total, 325 differential metabolites between the IFS and FFS were significantly (p < 0.05) enriched in the biosynthetic pathways of sesquiterpenes and triterpenes, as well as the ethylbenzene metabolic pathway. In total, 31 differentially expressed genes (DEGs) were related to terpenoid biosynthesis. There were only two DEGs related to the ethylbenzene metabolic pathway. In summary, we identified the volatile components of chestnut flowers and analyzed the changes in the contents of major volatile compounds in the flowers and the expression patterns of the related genes. The research results are helpful for understanding the regulation of VOCs in chestnut flowers.

Overgrazing typically leads to grassland vegetation degradation and reduction, which in turn triggers a series of ecological problems. Therefore, it is crucial to understand the effects of different Grazing Intensities (GIs) on the Vegetation Ecosystem (VE) to achieve sustainable grazing development. This study proposes a new quantitative index, the Grazing Intensity and Vegetation Cover Harmonization Index (GVCI), based on multiple indicators such as fractional vegetation cover (FVC), net primary productivity (NPP), and GI. The GVCI was used to quantify the “Harmonization and Conflict” status between GI and the VE in 39 Prefecture-Level Cities (PLCs) of the Qinghai-Tibet Plateau (QTP) and to evaluate the sustainable development level of grazing in different regions. In addition, the Random Forest (RF) model was used to simulate the GVCI development trend of various PLCs from 2015 to 2040. The results showed the following: (1) The GVCI can effectively quantify the response relationship between GI and the VE. The overall GVCI of the QTP was in the “Harmonization” state, with the proportion of areas in the “Harmonization” state fluctuating upwards. (2) The level of economic development intuitively affects the harmonization between grazing and the VE. Gross Domestic Product (GDP) is one of the important indicators of economic development level. PLCs with higher GDP levels exhibited a strong positive correlation between the GVCI and regional GDP. (3) The simulation results indicate that an increasing number of PLCs on the QTP will shift toward a “Harmonization” state. However, some PLCs in the western regions were still in an “Overload” state, and there is a need for close monitoring of their grazing activities and VE dynamics. The GVCI proposed in this study provides a novel methodology for quantifying the complex relationship between GI and the VE. It offers important scientific support for the sustainable development of grazing in ecologically fragile areas such as the QTP. The research results can be a robust scientific basis for the government to formulate reasonable grazing plans.

Background Silencing of the periostin gene ( POSTN ) can inhibit the biological process of several different cancers, and this inhibition may be related to down-regulation of PI3K/AKT signaling. However, the effect of POSTN on the progression, proliferation, and invasion of osteosarcoma (OS) remain unclear. Methods We used the Gene Expression Omnibus (GEO) database to screen datasets on in situ OS and lung metastases to identify core genes and potential pathways. We used additional bioinformatics tools to identify protein–protein interactions (PPIs) and gene networks, and selected the top seven genes whose expression had the strongest correlations with other genes. Results The results indicated that POSTN was a major hub gene. Subsequent analysis of gene expression profiles showed that POSTN was highly expressed in 262 cases with sarcoma and expression was closely related to poor prognosis. We also performed enrichment analysis to identify differentially expressed genes and used real-time PCR, western blotting, and immunohistochemistry analyses to measure POSTN expression in cells and tissues. Transfection of a POSTN -shRNA plasmid into cultured OS cells (Saos-2) effectively inhibited the proliferation, invasion, and migration of these cells. Taken together, our results suggest that POSTN may play a role in promoting the proliferation and metastasis of OS by activation of the PI3K/Akt signaling pathway. Conclusions Our results provide a preliminary characterization of the mechanism by which POSTN may regulate the migration and invasion of OS cells and also provide a theoretical basis for identifying biomarkers that have potential use for the diagnosis and treatment of OS.

The complete chloroplast genome sequence of a wild kiwifruit species from north China, Actinidia kolomikta , was determined and mapped based on Illumina sequencing data. The complete chloroplast genome is 156,875 bp and comprises a pair of inverted repeat regions of 23,918 bp each, a large single-copy region of 88,568 bp, and a small single-copy region of 20,471 bp. The genome harbors 132 genes, including 85 protein-coding genes, 8 ribosomal RNA genes, and 39 transfer RNA genes. Compared with most other angiosperms, A. kolomikta has lost its clpP gene. Phylogenetic analysis based on chloroplast genomes indicates that A. kolomikta is closely related to Actinidia argute .

The complete chloroplast genome sequence of a wild chestnut species, Castanea mollissima Blume, was determined and mapped based on Illumina sequencing data. The complete chloroplast genome is 160,869 bp and comprises a pair of inverted repeat regions of 25,701 bp each, a large single-copy region of 90,497 bp, and a small single-copy region of 18,970 bp. The genome harbors 131 genes, including 86 protein-coding genes, 8 ribosomal RNA genes, and 37 transfer RNA genes. Phylogenetic analysis based on chloroplast genomes indicates that wild C. mollissima is closely related to Castanopsis echinocarpa and dissimilar to cultivated C. mollissima .