Data reliability of the emerging citizen science in the Greater Bay Area of China

The potential of citizen science projects in research has been increasingly acknowledged, but the substantial engagement of these projects is restricted by the quality of citizen science data. Based on the largest emerging citizen science project in the country—Birdreport Online Database (BOD), we examined the biases of birdwatching data from the Greater Bay Area of China. The results show that the sampling effort is disparate among land cover types due to contributors' preference towards urban and suburban areas, indicating the environment suitable for species existence could be underrepresented in the BOD data. We tested the contributors' skill of species identification via a questionnaire targeting the citizen birders in the Greater Bay Area. The questionnaire show that most citizen birdwatchers could correctly identify the common species widely distributed in Southern China and the less common species with conspicuous morphological characteristics, while failed to identify the species from Alaudidae, Caprimulgidae, Emberizidae, Phylloscopidae, Scolopacidae and Scotocercidae. With a study example, we demonstrate that spatially clustered birdwatching visits can cause underestimation of species richness in insufficiently sampled areas; and the result of species richness mapping is sensitive to the contributors' skill of identifying bird species. Our results address how avian research can be influenced by the reliability of citizen science data in a region of generally high accessibility, and highlight the necessity of pre-analysis scrutiny on data reliability regarding to research aims at all spatial and temporal scales. To improve the data quality, we suggest to equip the data collection frame of BOD with a flexible filter for bird abundance, and questionnaires that collect information related to contributors’ bird identification skill. Statistic modelling approaches are encouraged to apply for correcting the bias of sampling effort

Functional Analysis of BcSNX3 in Regulating Resistance to Turnip Mosaic Virus (TuMV) by Autophagy in Pak-choi (Brassica campestris ssp. chinensis)

Sorting nexin protein is a class of highly conserved eukaryotic proteins containing the PX domain. Recent studies related to SNX in plants have focused on the regulation of abiotic stress processes, and there are few studies on the involvement of SNX in biological stress processes in plants. In this paper, a YTH assay and BiFC experiments were conducted twice to show that BcSNX3 (Brassica campestris Sorting nexin 3) interacted with CP and VPg of TuMV, and the interaction between BcATG8h (Brassica campestris autophagy-related gene 8h) and BcSNX3 was also found by YTH and BiFC. The colocalization of BcSNX3 and BcATG8b (Brassica campestris autophagy-related gene 8b) revealed BcSNX3 and autophagosome at the same place in the cell. QRT-PCR analysis showed that TuMV infection promotes the expression of BcSNX3, and the overexpression of this gene hinders the expression of autophagy-related genes and facilitates TuMV infection. VIGS was used to repress the expression level of the BcSNX3 gene in pak-choi to further study the function of BcSNX3 in the infection process of TuMV. After inoculation with TuMV, it was found that the accumulation of viral RNA in BcSNX3-gene-silenced plants was significantly less than in control plants. The accumulation of TuMV virus in the Arabidopsis snx3 knockout mutant was also less than in the wild type after TuMV inoculation. These results suggest that TuMV infection facilitates the expression of BcSNX3, and this gene may promote virus infection by inhibiting autophagy degradation of the virus and interacting with the CP and VPg of the virus. These results lay the foundation for the TuMV resistance breeding of pak-choi

A Quick Classifying Method for Tracking and Erosion Resistance of HTV Silicone Rubber Material via Laser-Induced Breakdown Spectroscopy

Silicone rubber material is widely used in high-voltage external insulation systems due to its excellent hydrophobicity and hydrophobicity transfer performance. However, silicone rubber is a polymeric material with a poor ability to resist electrical tracking and erosion; therefore, some fillers must be added to the material for performance enhancement. The inclined plane test is a standard method used for evaluating the tracking and erosion resistance by subjecting the materials to a combination of voltage stress and contaminate droplets to produce failure. This test is time-consuming and difficult to apply in field inspection. In this paper, a new and faster way to evaluate the tracking and erosion resistance performance is proposed using laser-induced breakdown spectroscopy (LIBS). The influence of filler content on the tracking and erosion resistance performance was studied, and the filler content was characterized by thermogravimetric analysis and the LIBS technique. In this paper, the tracking and erosion resistance of silicone rubber samples was correctly classified using principal component analysis (PCA) and neural network algorithms based on LIBS spectra. The conclusions of this work are of great significance to the performance characterization of silicone rubber composite materials

Integrating Dynamic 3D Chromatin Architecture and Gene Expression Alterations Reveal Heterosis in <i>Brassica rapa</i>

Heterosis plays a significant role in enhancing variety, boosting yield, and raising economic value in crops, but the molecular mechanism is still unclear. We analyzed the transcriptomes and 3D genomes of a hybrid (F1) and its parents (w30 and 082). The analysis of the expression revealed a total of 485 specially expressed genes (SEGs), 173 differentially expressed genes (DEGs) above the parental expression level, more actively expressed genes, and up-regulated DEGs in the F1. Further study revealed that the DEGs detected in the F1 and its parents were mainly involved in the response to auxin, plant hormone signal transduction, DNA metabolic process, purine metabolism, starch, and sucrose metabolism, which suggested that these biological processes may play a crucial role in the heterosis of Brassica rapa. The analysis of 3D genome data revealed that hybrid F1 plants tend to contain more transcriptionally active A chromatin compartments after hybridization. Supplementaryly, the F1 had a smaller TAD (topologically associated domain) genome length, but the number was the highest, and the expression change in activated TAD was higher than that of repressed TAD. More specific TAD boundaries were detected between the parents and F1. Subsequently, 140 DEGs with genomic structural variants were selected as potential candidate genes. We found two DEGs with consistent expression changes in A/B compartments and TADs. Our findings suggested that genomic structural variants, such as TADs and A/B chromatin compartments, may affect gene expression and contribute to heterosis in Brassica rapa. This study provides further insight into the molecular mechanism of heterosis in Brassica rapa

Genome-wide analysis of auxin transport genes identifies the hormone responsive patterns associated with leafy head formation in Chinese cabbage

Auxin resistant 1/like aux1 (AUX/LAX), pin-formed (PIN) and ATP binding cassette subfamily B (ABCB/MDR/PGP) are three families of auxin transport genes. The development-related functions of the influx and efflux carriers have been well studied and characterized in model plants. However, there is scant information regarding the functions of auxin genes in Chinese cabbage and the responses of exogenous polar auxin transport inhibitors (PATIs). We conducted a whole-genome annotation and a bioinformatics analysis of BrAUX/LAX, BrPIN, and BrPGP genes in Chinese cabbage. By analyzing the expression patterns at several developmental stages in the formation of heading leaves, we found that most auxin-associate genes were expressed throughout the entire process of leafy head formation, suggesting that these genes played important roles in the development of heads. UPLC was used to detect the distinct and uneven distribution of auxin in various segments of the leafy head and in response to PATI treatment, indicated that the formation of the leafy head depends on polar auxin transport and the uneven distribution of auxin in leaves. This study provides new insight into auxin polar transporters and the possible roles of the BrLAX, BrPIN and BrPGP genes in leafy head formation in Chinese cabbage

Synthesis and Biological Evaluation of Sclareolide-Indole Conjugates and Their Derivatives

Sclareolide is a sesquiterpene lactone isolated from various plant sources in tons every year and is commercially used as a flavor ingredient in the cosmetic and food industries. Antitumor and antiviral activities of sclareolide have been previously reported. However, biological studies of sclareolide synthetic analogous are few. In view of these, we developed a robust synthetic method that allows the assembly of 36 novel sclareolide-indole conjugates and their derivatives. The synthetic method was based on TiCl4-promoted nucleophilic substitution of sclareolide-derived hemiacetal 4, while electron-rich aryles including indoles, polyphenol ethers, and pyrazolo [1,5-a]pyridine were good substrates. The stereochemistry of the final products was confirmed by single-crystal X-ray diffraction analysis, while the antiproliferative activities of selected final products were tested in K562 and MV4-11 cancer cell lines. Cytometric flow analysis shows that lead compounds 8k- and 10-induced robust apoptosis in MV4-11 cancer cells, while they exhibited weak impact on cell cycle progression. Taken together, our study suggests that sclareolide could be a good template and substrate for the synthesis of novel antiproliferative compounds

Host Factors Genes <i>BcCLC1</i> and <i>BcCLC2</i> Confer Turnip Mosaic Virus Resistance in Non-Heading Chinese Cabbage (<i>Brassica campestris</i> ssp. <i>chinensis</i>)

Clathrin is an evolutionarily highly conserved evolutionary protein consisting of clathrin light chains (CLC) and clathrin heavy chains (CHC), and these form its basic structure. Clathrin is an important host factor in the process of viral infection. In this study, we cloned the BcCLC1 gene and the BcCLC2 gene from the ‘49CX’ variety of non-heading Chinese cabbage (NHCC, Brassica campestris L. ssp. chinensis Makino) and verified their functions. The results showed that BcCLC1 was mainly localized in the cytomembrane and cytoplasm, and only a small amount entered the nucleus. BcCLC2 encoded a protein comprising 265 amino acids that were distributed in the cytomembrane, nucleus, and cytoplasm. A BiFC assay and yeast two-hybrid (Y2H) analysis showed that BcCLCs (BcCLC1 and BcCLC2) could interact with several TuMV proteins. We further investigated the mechanism of BcCLCs in regulating TuMV virus infections in NHCC, and observed that BcCLCs gene silencing inhibited TuMV infections and overexpression of BcCLCs in Arabidopsis promoted TuMV infections in NHCC. Finally, mutants of Arabidopsis homologs of BcCLCs were also screened and subjected to TuMV inoculation tests. In conclusion, we speculate that BcCLCs confer Turnip mosaic virus (TuMV) resistance in NHCC by interacting with TuMV proteins to promote the intracellular transport of the virus

Mechano-bioconjugation Strategy Empowering Fusion Protein Therapeutics with Aggregation Resistance, Prolonged Circulation, and Enhanced Antitumor Efficacy

Bioconjugation is a powerful protein modification strategy to improve protein properties. Herein, we report mechano-bioconjugation as a novel approach to empower fusion protein therapeutics and demonstrate its utility by a protein heterocatenane (cat-IFN-ABD) containing interferon-alpha 2b (IFN) mechanically interlocked with a consensus albumin-binding domain (ABD). The conjugate was selectively synthesized in cellulo following a cascade of post-translational events using a pair of heterodimerizing p53dim variants and two orthogonal split-intein reactions. The catenane topology was proven by combined techniques of LC-MS, SDS-PAGE, SEC, and controlled proteolytic digestion. Not only did caiIFN-ABD retain activities comparable to those of the wild-type IFN and ABD, the conjugate also exhibited enhanced aggregation resistance and prolonged circulation time over the simple linear and cyclic fusions. Consequently, cat-IFN-ABD potently inhibited tumor growth in the mouse xenograft model. Therefore, mechano-bioconjugation by catenation accomplishes function integration with additional benefits, providing an alternative pathway for developing advanced protein therapeutics

Mechano-bioconjugation Strategy Empowering Fusion Protein Therapeutics with Aggregation Resistance, Prolonged Circulation, and Enhanced Antitumor Efficacy

Bioconjugation is a powerful protein modification strategy to improve protein properties. Herein, we report mechano-bioconjugation as a novel approach to empower fusion protein therapeutics and demonstrate its utility by a protein heterocatenane (cat-IFN-ABD) containing interferon-alpha 2b (IFN) mechanically interlocked with a consensus albumin-binding domain (ABD). The conjugate was selectively synthesized in cellulo following a cascade of post-translational events using a pair of heterodimerizing p53dim variants and two orthogonal split-intein reactions. The catenane topology was proven by combined techniques of LC-MS, SDS-PAGE, SEC, and controlled proteolytic digestion. Not only did caiIFN-ABD retain activities comparable to those of the wild-type IFN and ABD, the conjugate also exhibited enhanced aggregation resistance and prolonged circulation time over the simple linear and cyclic fusions. Consequently, cat-IFN-ABD potently inhibited tumor growth in the mouse xenograft model. Therefore, mechano-bioconjugation by catenation accomplishes function integration with additional benefits, providing an alternative pathway for developing advanced protein therapeutics