Dynamic placement of the linker histone H1 associated with nucleosome arrangement and gene transcription in early Drosophila embryonic development

The linker histone H1 is critical to maintenance of higher-order chromatin structures and to gene expression regulation. However, H1 dynamics and its functions in embryonic development remain unresolved. Here, we profiled gene expression, nucleosome positions, and H1 locations in early Drosophila embryos. The results show that H1 binding is positively correlated with the stability of beads-on-a-string nucleosome organization likely through stabilizing nucleosome positioning and maintaining nucleosome spacing. Strikingly, nucleosomes with H1 placement deviating to the left or the right relative to the dyad shift to the left or the right, respectively, during early Drosophila embryonic development. H1 occupancy on genic nucleosomes is inversely correlated with nucleosome distance to the transcription start sites. This inverse correlation reduces as gene transcription levels decrease. Additionally, H1 occupancy is lower at the 5\u27 border of genic nucleosomes than that at the 3\u27 border. This asymmetrical pattern of H1 occupancy on genic nucleosomes diminishes as gene transcription levels decrease. These findings shed new lights into how H1 placement dynamics correlates with nucleosome positioning and gene transcription during early Drosophila embryonic development

Kinematic analysis and process optimization of root-cutting systems in field harvesting of garlic based on computer simulation technology

IntroductionRoot cutting is an important process in garlic field harvesting but is the weakest link in the full mechanization of garlic production. To improve the current situation of technological backwardness and poor operational quality of mechanized garlic root-cutting in the main garlic-producing regions of China, this study combined the physical characteristics and agronomic requirements of garlic plants, and proposed an innovative floating root-cutting technology for garlic combine harvesters that enables the top alignment of bulb, adaptive profiling floating of cutter, and embedded cutting of roots.MethodsThrough the kinematic analysis of the floating cutting process, the coordinate equations of the initial contact point of the bulb, the mathematical model of the floating displacement of the cutting component. Using computer simulation techniques, the dynamic simulation study of the floating cutting process was carried out in the rigid-flexible coupling numerical simulation model of root-cutting mechanism and garlic plant. The influence law of garlic conveying speed, extension spring preload force and stiffness on the floating displacement of the cutting component and the angular velocity of swing arm reset and its formation causes were analyzed by a single-factor simulation test. The key operating parameters of the root-cutting mechanism were optimized through the computerized virtual orthogonal test and fuzzy comprehensive evaluation.Results and discussionThe significance of the factors affecting the floating cutting performance decreased in the following order: extension spring preload force, garlic conveying speed and extension spring stiffness. The optimal parameter combination of the root cutting mechanism obtained from the optimization were as follow: extension spring preload force was 16 N, garlic conveying speed was 0.8 m/s, and extension spring stiffness was 215 N/m. Tests conducted with the optimal parameter combination yielded a root excision rate of 92.72%, which meets the requirements of Chinese garlic field harvesting quality. This study provides computer simulation optimization methods for the optimal design of the root-cutting mechanism, and also provides technical and equipment support for the full mechanization of garlic production in China

Complex origins indicate a potential bridgehead introduction of an emerging amphibian invader (Eleutherodactylus planirostris) in China

Identifying the origins of established alien species is important to prevent new introductions in the future. The greenhouse frog (Eleutherodactylus planirostris), native to Cuba, the Bahamas, and the Cayman Islands, has been widely introduced to the Caribbean, North and Central America, Oceania and Asia. This invasive alien amphibian was recently reported in Shenzhen, China, but the potential introduction sources remain poorly understood. Based on phylogenetic analysis using mitochondrial 16S, COI and CYTB sequences, we detected a complex introduction origin of this species, which may be from Hong Kong, China, the Philippines, Panama and Florida, USA, all pointing to a bridgehead introduction. In addition, the nursery trade between the four countries or regions and mainland China from 2011 to 2020 was also significantly higher than other areas with less likelihood of introductions, which supported the molecular results. Our study provides the first genetic evidence of the potential sources of this emerging amphibian invader in mainland China, which may help develop alien species control strategies in the face of growing trade through globalization

Genome-wide identification of the soybean cytokinin oxidase/dehydrogenase gene family and its diverse roles in response to multiple abiotic stress

Cytokinin oxidase/dehydrogenase (CKX) irreversibly degrades cytokinin, regulates growth and development, and helps plants to respond to environmental stress. Although the CKX gene has been well characterized in various plants, its role in soybean remains elusive. Therefore, in this study, the evolutionary relationship, chromosomal location, gene structure, motifs, cis-regulatory elements, collinearity, and gene expression patterns of GmCKXs were analyzed using RNA-seq, quantitative real-time PCR (qRT-PCR), and bioinformatics. We identified 18 GmCKX genes from the soybean genome and grouped them into five clades, each comprising members with similar gene structures and motifs. Cis-acting elements involved in hormones, resistance, and physiological metabolism were detected in the promoter regions of GmCKXs. Synteny analysis indicated that segmental duplication events contributed to the expansion of the soybean CKX family. The expression profiling of the GmCKXs genes using qRT-PCR showed tissue-specific expression patterns. The RNA-seq analysis also indicated that GmCKXs play an important role in response to salt and drought stresses at the seedling stage. The responses of the genes to salt, drought, synthetic cytokinin 6-benzyl aminopurine (6-BA), and the auxin indole-3-acetic acid (IAA) at the germination stage were further evaluated by qRT-PCR. Specifically, the GmCKX14 gene was downregulated in the roots and the radicles at the germination stage. The hormones 6-BA and IAA repressed the expression levels of GmCKX1, GmCKX6, and GmCKX9 genes but upregulated the expression levels of GmCKX10 and GmCKX18 genes. The three abiotic stresses also decreased the zeatin content in soybean radicle but enhanced the activity of the CKX enzymes. Conversely, the 6-BA and IAA treatments enhanced the CKX enzymes’ activity but reduced the zeatin content in the radicles. This study, therefore, provides a reference for the functional analysis of GmCKXs in soybean in response to abiotic stresses

Characterization of Non-heading Mutation in Heading Chinese Cabbage (Brassica rapa L. ssp. pekinensis)

Heading is a key agronomic trait of Chinese cabbage. A non-heading mutant with flat growth of heading leaves (fg-1) was isolated from an EMS-induced mutant population of the heading Chinese cabbage inbred line A03. In fg-1 mutant plants, the heading leaves are flat similar to rosette leaves. The epidermal cells on the adaxial surface of these leaves are significantly smaller, while those on the abaxial surface are much larger than in A03 plants. The segregation of the heading phenotype in the F2 and BC1 population suggests that the mutant trait is controlled by a pair of recessive alleles. Phytohormone analysis at the early heading stage showed significant decreases in IAA, ABA, JA and SA, with increases in methyl IAA and trans-Zeatin levels, suggesting they may coordinate leaf adaxial-abaxial polarity, development and morphology in fg-1. RNA-sequencing analysis at the early heading stage showed a decrease in expression levels of several auxin transport (BrAUX1, BrLAXs, and BrPINs) and responsive genes. Transcript levels of important ABA responsive genes, including BrABF3, were up-regulated in mid-leaf sections suggesting that both auxin and ABA signaling pathways play important roles in regulating leaf heading. In addition, a significant reduction in BrIAMT1 transcripts in fg-1 might contribute to leaf epinastic growth. The expression profiles of 19 genes with known roles in leaf polarity were significantly different in fg-1 leaves compared to wild type, suggesting that these genes might also regulate leaf heading in Chinese cabbage. In conclusion, leaf heading in Chinese cabbage is controlled through a complex network of hormone signaling and abaxial-adaxial patterning pathways. These findings increase our understanding of the molecular basis of head formation in Chinese cabbage

The Arg233Lys AQP0 Mutation Disturbs Aquaporin0-Calmodulin Interaction Causing Polymorphic Congenital Cataract

Calmodulin (CaM) directly interacts with the aquaporin 0 (AQP0) C-terminus in a calcium dependent manner to regulate the water permeability of AQP0. We previously identified a missense mutation (p.R233K) in the putative CaM binding domain of AQP0 C-terminus in a congenital cataract family. This study was aimed at exploring the potential pathogenesis of this mutation causative of cataract and mainly identifying how it influenced the binding of AQP0 to CaM. Wild type and R233K mutant AQP0 with EGFP-tag were transfected separately into Hela cells to determine the expression and subcellular localizations. The co-immunoprecipitation (CoIP) assay was used to detect the interaction between AQP0 and CaM. AQP0 C-terminus peptides were synthesized with and without R233K, and the binding abilities of these peptides to CaM were assessed using a fluorescence binding assay. Localizations of wild type and R233K mutant AQP0 were determined from EGFP fluorescence, and the chimeric proteins were both localized abundantly in the plasma membrane. Protein expression levels of the culture cells showed no significant difference between them. The results from CoIP assay implied that R233K mutant presented more weakly in association with CaM than wild type AQP0. The AQP0 C-terminal mutant peptide was found to have 2.5-fold lower binding affinity to CaM than wild type peptide. These results suggested that R233K mutation did not affect the expression, location and trafficking of the protein but did influence the interaction between AQP0 and CaM. The binding affinity of AQP0 C-terminus to CaM was significantly reduced. Due to lack of the modulation of the Ca2+-calmodulin complex, the water permeability of AQP0 was subsequently augmented, which might lead to the development of this cataract