Integrative transcriptome analysis uncovers common components containing CPS2 regulated by maize lncRNA GARR2 in gibberellin response.

MAIN CONCLUSION: The combined transcriptome outcome provides an important clue to the regulatory cascade centering on lncRNA GARR2 and CPS2 gene in GA response. Long non-coding RNAs (lncRNAs) serve as regulatory components in transcriptional hierarchy governing multiple aspects of biological processes. Dissecting regulatory mechanisms underpinning tetracyclic diterpenoid gibberellin (GA) cascade holds both theoretical and applied significance. However, roles of lncRNAs in transcriptional modulation of GA pathway remain largely elusive. Gypsy retrotransposon-derived GIBBERELLIN RESPONSIVE lncRNA2 (GARR2) has been reported as GA-responsive maize lncRNA. Here a novel GARR2-edited line garr2-1 was identified, characteristic of GA-induced phenotype of increased seedling height and elongated leaf sheath. Transcriptome analysis indicated that transcriptional abundance of five genes [ent-copalyl diphosphate synthase2 (CPS2), ent-kaurene synthase4 (KS4), ent-kaurene synthase6 (KS6), ent-kaurene oxidase2 (KO2), and ent-kaurenoic acid oxidase1/Dwarf3 (KAO1/D3)] was elevated in garr2-1 for early steps of GA biosynthesis. Five GA biosynthetic genes as hub regulators were interlaced to shape regulatory network of GA response. Different transcriptome resources were integrated to discover common differentially expressed genes (DEGs) in the independent GARR2-edited lines GARR2KO and garr2-1. A total of 320 common DEGs were retrieved. These common DEGs were enriched in diterpenoid biosynthetic pathway. Integrative transcriptome analysis revealed the common CPS2 encoding the CPS enzyme that catalyzes the conversion of the precursor trans-geranylgeranyl diphosphate to ent-copalyl diphosphate. The up-regulated CPS2 supported the GA-induced phenotype of slender seedlings observed in the independent GARR2-edited lines GARR2KO and garr2-1. Our integrative transcriptome analysis uncovers common components of the GA pathway regulated by lncRNA GARR2. These common components, especially for the GA biosynthetic gene CPS2, provide a valuable resource for further delineating the underlying mechanisms of lncRNA GARR2 in GA response.

In vivo transgenic studies confirm the critical acylation function of LeBAHD56 for shikonin in Lithospermum erythrorhizon.

KEY MESSAGE: LeBAHD56 is preferentially expressed in tissues where shikonin and its derivatives are biosynthesized, and it confers shikonin acylation in vivo. Two WRKY transcriptional factors might regulate LeBAHD56's expression. Shikonin and its derivatives, found in the roots of Lithospermum erythrorhizon, have extensive application in the field of medicine, cosmetics, and other industries. Prior research has demonstrated that LeBAHD1(LeSAT1) is responsible for the biochemical process of shikonin acylation both in vitro and in vivo. However, with the exception of its documented in vitro biochemical function, there is no in vivo genetic evidence supporting the acylation function of the highly homologous gene of LeSAT1, LeBAHD56(LeSAT2), apart from its reported role. Here, we validated the critical acylation function of LeBAHD56 for shikonin using overexpression (OE) and CRISPR/Cas9-based knockout (KO) strategies. The results showed that the OE lines had a significantly higher ratio of acetylshikonin, isobutyrylshikonin or isovalerylshikonin to shikonin than the control. In contrast, the KO lines had a significantly lower ratio of acetylshikonin, isobutyrylshikonin or isovalerylshikonin to shikonin than controls. As for its detailed expression patterns, we found that LeBAHD56 is preferentially expressed in roots and callus cells, which are the biosynthesis sites for shikonin and its derivatives. In addition, we anticipated that a wide range of putative transcription factors might control its transcription and verified the direct binding of two crucial WRKY members to the LeBAHD56 promoter's W-box. Our results not only confirmed the in vivo function of LeBAHD56 in shikonin acylation, but also shed light on its transcriptional regulation.

Sugar transporter ZmSWEET1b is responsible for assimilate allocation and salt stress response in maize.

Sugar efflux transporter SWEET family is involved in multiple biological processes, from nectar secretion, pollen fertility to seed filling. Although roles of SWEETs in abiotic stress adaption have been revealed mainly in reference organism Arabidopsis, cereal crops SWEETs responses to abiotic stimulation remain largely elusive. Here, we report the characterization of maize SWEET family member ZmSWEET1b, with emphasis on its response to salinity stress. ZmSWEET1b is a canonical sugar transporter, characteristic of seven transmembrane helices and plasma membrane localization. ZmSWEET1b and its rice ortholog OsSWEET1b in phylogenetic clade I underwent convergent selection during evolution. Two independent knockout lines were created by the CRISPR/Cas9 method to functionally characterized ZmSWEET1b. Sucrose and fructose contents are significantly decreased in ZmSWEET1b knockout lines. Mature leaves of ZmSWEET1b-edited lines exhibit chlorosis, reminiscent of senescence-like phenotype. Ears and seeds of ZmSWEET1b knockout lines are small. Upon salinity treatment, ZmSWEET1b-edited lines become more wilted. Transcriptional abundance of genes for Na+ efflux from roots to the rhizosphere, including ZmSOS1, ZmH+-ATPASE 2, and ZmH+-ATPASE 8, is decreased in salt-treated ZmSWEET1b knockout lines. These findings indicate that convergently selected sugar transporter ZmSWEET1b is important for maize plant development and responses to salt stress. The manipulation of ZmSWEET1b may represent a feasible way forward in the breeding of salinity tolerant ideotypes through the optimization of assimilate allocation.

Large scanning field laser concurrent drilling system with a five-axis independent control for special-shaped holes.

Five-axis laser scanning technology is an effective drilling method for special-shaped holes. Due to a gap in laser angle-of-incidence (AOI) control within a large scanning field, current technologies are challenging for fabricating large-size holes or special-shaped hole arrays. In this paper, a large scanning field five-axis laser concurrent drilling system was proposed. The laser AOI was independently controlled using two pairs of synchronous deflection mirrors. The laser control deviations under a large scanning field were investigated systematically by simulation and experiment. By establishing a complete correction method, the laser AOI control within a scanning field diameter of up to 35 mm was achieved. A series of special-shaped holes were fabricated concurrently on a 3.6 mm thick glass fiber reinforced plastic (GFRP), verifying that the AOI can be controlled by the five-axis laser scanning system. Our work provides a novel method to increase the scanning field of the five-axis laser scanning technology, expanding the application scope of the five-axis laser processing.

Extending the 3D scanning range of reflective dynamic focusing device-based laser scanners.

Reflective dynamic focusing devices (RDFDs) have shown their potential in laser scanning as high-performance laser Z-direction focusing devices. However, the scanning range of RDFD-based scanners is limited by aberrations during dynamic focusing. An aspheric symmetry correction (ASC) method was proposed to extend the effective scanning range. An aspheric lens was introduced to correct the optical path difference (OPD) and optimize aberrations. As a result, the scanning range in the three-dimensional (3D) space increased by 15.2%. The ASC method has been proven to extend the 3D scanning range of RDFD-based scanners and may have broad application prospects.

A Scalable Cross-Chain Access Control and Identity Authentication Scheme.

Cross-chain is an emerging blockchain technology which builds the bridge across homogeneous and heterogeneous blockchains. However, due to the differentiation of different blockchains and the lack of access control and identity authentication of cross-chain operation subjects, existing cross-chain technologies are struggling to accomplish the identity transformation of cross-chain subjects between different chains, and also pose great challenges in terms of the traceability and supervision of dangerous transactions. To address the above issues, this paper proposes a scalable cross-chain access control and identity authentication scheme, which can authenticate the legitimacy of blockchains in the cross-chain system and ensure that all cross-chain operations are carried out by verified users. Furthermore, it will record all cross-chain operations with the help of Superchain in order to regulate and trace illegal transactions. Our scheme is scalable and, at the same time, has low invasiveness to blockchains in the cross-chain system. We implement the scheme and accordingly conduct the evaluations, which prove its security, efficiency, and scalability.

G-protein couples MAPK cascade through maize heterotrimeric Gß subunit.

KEY MESSAGE: G protein couples MAPK cascade through maize heterotrimeric Gß subunit MGB1. Heterotrimeric G protein Gß interacts with Gγ subunit to generate Gßγ dimer in modulation of various biological processes. The modulatory events at transcriptome scale of plant Gß subunit remain largely unknown. To reveal the regulatory basis of Gß subunit at transcriptome level, we first identified a canonical maize Gß subunit MGB1 that physically interacted with Type C Gγ protein MGG4. For transcriptome analysis, two independent CRISPR/Cas9-edited MGB1 lines were generated, which all exhibited growth arrest, suggestive of MGB1 essential for maize seedling establishment. Transcriptomic outcomes showed that MGB1 knockout resulted in elevated transcriptional abundance of plant immune response marker PR and immune receptor RPM1. Integrated GO, KEGG, and GSEA analyses pinpointed the enrichment of differentially expressed genes in defense response pathway. Functional association network construction revealed MAPK cascade components and PR protein as hub regulators of MGB1-mediated immune signaling. MGB1 and scaffold protein ZmRACK1 together with MAPK cascade components coordinately modulated maize immune responses. We built a modulatory hierarchy of Gß subunit at transcriptome and interacting scales, which is informative for our understanding of the regulatory basis of G protein signaling.

The Effects of Lentinan on the Hematological and Immune Indices of Dairy Cows.

In this study, we investigated the effects of lentinan (LNT) on hematological parameters, immune indices, and metabolite levels in dairy cows. We randomly assigned forty Holstein cows to four treatment groups. The treatments consisted of 0, 5, 10, and 15 g/d of LNT. Compared with the control group, the addition of 10 g/d of LNT decreased the content of ALT and IL-8 but simultaneously increased the content of IL-4 in the cows' serum. Supplementation with 10 g/d of LNT decreased the levels of lymphocyte, RDW, ALT, AST, TC, IL-2, and IL-8, but, concurrently, in-creased the levels of granulocytes and IL-4 in their serum. In addition, supplementation with 15 g/d of LNT decreased the levels of RDW, TC, IL-2, and IL-8, but, at the same time, increased the levels of IL-4 and IgM in their serum. For the metabolomic analysis, cows fed with 0 and 10 g/d of LNT were selected. The results showed that 10 metabolites, including reduced nicotinamide riboside and trehalose, were upregulated in the 10 g/d group. These differential metabolites were enriched in tyrosine metabolism and trehalose degradation and altered two metabolic pathways of ubiquinone and other terpene quinone biosynthesis, as well as starch and sucrose metabolism. These findings provide evidence that LNT could be used to reduce the risk of inflammation in dairy cows.

Analysis of spatiotemporal patterns of atmospheric CO2 concentration in the Yellow River Basin over the past decade based on time-series remote sensing data.

Understanding the spatial and temporal variations of CO2 column concentration (CO2-CCs) is crucial for tackling climate change and promoting sustainable human development. This study provides an in-depth analysis of CO2 dynamics in the Yellow River Basin, an area significantly affected by both natural and anthropogenic factors. Using data from the Orbiting Carbon Observatory 2 (OCO-2) and the Fourier transformation spectrometer (FTS) of the GOSAT satellite remote sensing sensors, supplemented with ground station data from the Waliguan station, we scrutinized the CO2 levels in the region from 2013 to 2022. The regional CO2-CC displayed a 12-month cyclical variation and a continuous upward trend, escalating by approximately 4.26% over the 10-year period. Spatiotemporal differences were evident in the monthly variation of CO2-CC, with peak and minimum values occurring in May and August respectively. Geographically, the highest CO2-CC was found in the central part of the basin, while the lowest was in the northern part of Inner Mongolia. This study underscores the increased significance of the region's CO2-CC, which showed an increase from 17.0 ppm at the start of the period to 21.0 ppm by the end, representing an overall growth of between 4.35 and 5.25%. The findings highlight the urgency of targeted measures to mitigate CO2 emissions and adapt to their consequences in the Yellow River Basin, contributing to the global efforts against climate change and towards sustainable development.

A Comparative Study on Microstructure and Properties of Ultra-High-Speed Laser Cladding and Traditional Laser Cladding of Inconel625 Coatings.

In this study, ultra-high-speed laser cladding (UHSLC) and traditional low-speed laser cladding (LSLC) were employed to prepare high-quality Inconel625 coatings on 27SiMn substrates. UHSLC has cladding speeds of 30 m/min, which are 15 times faster than those of LSLC, and it produces a much greater cladding efficiency, which is 13.9 times greater than LSLC. The microstructure of the Inconel625 coatings was investigated in detail utilizing field emission scanning electron microscopy (FESEM) and electron probe microanalyzer (EPMA). According to the FESEM results, UHSLC Inconel625 coatings have more refined crystals than LSLC Inconel625 coatings. Nevertheless, the EPMA results indicate that the UHSLC Inconel625 coatings exhibit much more severe elemental segregation. Moreover, the hardness, wear and corrosion resistance of Inconel625 coatings are significantly enhanced by increasing the laser cladding speed. Furthermore, the reasons for the differences in microstructure and properties of Inconel625 coatings prepared by UHSLC and LSLC were clarified by finite element simulation. UHSLC technique is, therefore, more suitable for preparing Inconel625 coatings on 27SiMn steel surfaces than LSLC.

Overexpressed LINC00467 promotes the viability and proliferation yet inhibits apoptosis of gastric cancer cells via raising ITGB3 level.

Long non-coding RNA (lncRNA) LINC00467 plays a proto-oncogenic role in non-small cell lung cancer. However, its effect and modulatory mechanism in gastric cancer (GC) are unknown. Thereby, we elucidated the mechanism of LINC00467 in GC. LINC00467 level in GC tissues was assessed by bioinformatic analysis, and clinicopathological parameters from GC patients were collected. The levels of LINC00467, integrin subunit beta 3 (ITGB3), proliferating cell nuclear antigen (PCNA), cleaved caspase-3 and cleaved poly (ADP-ribose) polymerase 1 (PARP1) in tissue samples or treated GC cells were assessed by quantitative real-time polymerase chain reaction (qRT-PCR), fluorescence in situ hybridization (FISH), or Western blot. The viability, proliferation and apoptosis of GC cells were detected by methyl thiazolyl tetrazolium assay, colony formation assay, and flow cytometry. Levels of LINC00467 and ITGB3 were up-regulated in GC, and highly expressed LINC00467 was positively associated with tumor size, differentiation, N stage, and T stage in GC patients. LINC00467 was enriched in cytoplasm of GC cells, and overexpressed LINC00467 promoted the viability and proliferation as well as levels of ITGB3 and PCNA, while suppressing the apoptosis and levels of cleaved caspase-3 and cleaved PARP1 in GC cells. Besides, the effects of shLINC00467 on inhibiting cell viability, proliferation of GC cells and PCNA level and promoting apoptosis as well as levels of cleaved caspase-3 and cleaved PARP1 were all partially reversed by overexpressed ITGB3. Overexpressed LINC00467 enhanced the viability and proliferation but inhibited apoptosis of GC cells via increasing ITGB3 level.