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Abstract Given the aggressive nature and robust survival capabilities of the alligator gar (Atractosteus spatula), if it was to exist in a new environment as an invasive species, it could cause significant disruption to the invaded ecosystem. Building on the continuity and completeness of the existing draft genome were not optimal, this study has updated a high-quality genome of the alligator gar at the chromosome level, which was assembled using Oxford Nanopore Technology and chromatin interaction mapping (Hi-C) sequencing techniques. In summary, the alligator gar genome in this study was 1.05 Gb in size with a contig N50 of 15.7 Mb and scaffold N50 of 56.8 Mb. We captured 98.26% of assembled bases in 28 pseudochromosomes. The completeness of the final chromosome-level genome reached 96.7%. Meanwhile, a total of 19,103 protein-coding genes were predicted, of which 99.83% could be predicted with functions. Taken together, the present high-quality alligator gar chromosome-level genome provides a valuable resource for exploring the underlying genomic basis to comprehend the functional genomics, chromosome evolution, and population management of this species.

Abstract Background Long non-coding RNAs (lncRNAs) play a crucial role in regulating gene expression vital for the growth and development of plants. Despite this, the role of lncRNAs in Chinese cabbage (Brassica rapa L. ssp. pekinensis) pollen development and male fertility remains poorly understood. Results In this study, we characterized a recessive genic male sterile mutant (366–2 S), where the delayed degradation of tapetum and the failure of tetrad separation primarily led to the inability to form single microspores, resulting in male sterility. To analyze the role of lncRNAs in pollen development, we conducted a comparative lncRNA sequencing using anthers from the male sterile mutant line (366–2 S) and the wild-type male fertile line (366–2 F). We identified 385 differentially expressed lncRNAs between the 366–2 F and 366–2 S lines, with 172 of them potentially associated with target genes. To further understand the alterations in mRNA expression and explore potential lncRNA-target genes (mRNAs), we performed comparative mRNA transcriptome analysis in the anthers of 366–2 S and 366–2 F at two stages. We identified 1,176 differentially expressed mRNAs. Remarkably, GO analysis revealed significant enrichment in five GO terms, most notably involving mRNAs annotated as pectinesterase and polygalacturonase, which play roles in cell wall degradation. The considerable downregulation of these genes might contribute to the delayed degradation of tapetum in 366–2 S. Furthermore, we identified 15 lncRNA-mRNA modules through Venn diagram analysis. Among them, MSTRG.9997-BraA04g004630.3 C (β-1,3-glucanase) is associated with callose degradation and tetrad separation. Additionally, MSTRG.5212-BraA02g040020.3 C (pectinesterase) and MSTRG.13,532-BraA05g030320.3 C (pectinesterase) are associated with cell wall degradation of the tapetum, indicating that these three candidate lncRNA-mRNA modules potentially regulate pollen development. Conclusion This study lays the foundation for understanding the roles of lncRNAs in pollen development and for elucidating their molecular mechanisms in regulating male sterility in Chinese cabbage.

Study region: The study region of this paper is the Wusuli River basin, which is comprised of nine sub-basins. The basin is located on the border between China and Russia. Study focus: We investigated the hydrological connectivity of the Wusuli River basin, an international boundary river that has received little attention. The spatial and temporal evolution of hydrological connectivity over the last 38 years was characterized using a geostatistical connectivity function combined with the remote sensing technique. In particular, some key sub-basins were identified to help strengthen China-Russian cooperation on ecological protection of the basin. New hydrological insights for the region: The results show that the hydrological connectivity in the northwestern region of the Wusuli River basin was unstable with several surges from 1984 to 2021. Analyzed from a temporal perspective, the effective range of hydrological connectivity is expanding and connectivity is showing an overall upward trend due to increased fluctuations in rainfall. Analyzed from a spatial perspective, the connectivity of the right bank of the Wusuli River is better than that of the left bank due to the influence of topography. Stable connectivity pathways are a must for efficient hydrological connectivity within a watershed. Another finding is that the capacity of the Wusuli River basin to resist extreme rainfall is weakening.

Clubroot disease poses a significant threat to Brassica crops, necessitating ongoing updates on resistance gene sources. In F2 segregants of the clubroot-resistant inbred line BrT18-6-4-3 and susceptible DH line Y510, the genetic analysis identified a single dominant gene responsible for clubroot resistance. Through bulk segregant sequencing analysis and kompetitive allele-specific polymerase chain reaction assays, CRA8.1.6 was mapped within 110 kb (12,255–12,365 Mb) between markers L-CR11 and L-CR12 on chromosome A08. We identified B raA08g015220.3.5C as the candidate gene of CRA8.1.6. Upon comparison with the sequence of disease-resistant material BrT18-6-4-3, we found 249 single-nucleotide polymorphisms, seven insertions, six deletions, and a long terminal repeat (LTR) retrotransposon (5,310 bp) at 909 bp of the first intron. However, the LTR retrotransposon was absent in the coding sequence of the susceptible DH line Y510. Given the presence of a non-functional LTR insertion in other materials, it showed that the LTR insertion might not be associated with susceptibility. Sequence alignment analysis revealed that the fourth exon of the susceptible line harbored two deletions and an insertion, resulting in a frameshift mutation at 8,551 bp, leading to translation termination at the leucine-rich repeat domain’s C-terminal in susceptible material. Sequence alignment of the CDS revealed a 99.4% similarity to Crr1a, which indicate that CRA8.1.6 is likely an allele of the Crr1a gene. Two functional markers, CRA08-InDel and CRA08-KASP1, have been developed for marker-assisted selection in CR turnip cultivars. Our findings could facilitate the development of clubroot-resistance turnip cultivars through marker-assisted selection.

Dissimilatory iron reduction (DIR) coupled with carbon cycling is increasingly being recognized as an influential process in freshwater wetland soils and sediments. The role of DIR in organic matter (OM) mineralization, however, is still largely unknown in lake sediment environments. In this study, we clarified rates and pathways of OM mineralization in two shallow lakes with seasonal hydrological connectivity and different eutrophic situations. We found that in comparison with the domination of DIR (55%) for OM mineralization in Lake Xiaoxingkai, the contribution of methanogenesis was much higher (68%) in its connected lake (Lake Xingkai). The differences in rates and pathways of sediment OM mineralization between the two lakes were attributed to higher concentrations of carbonate associated iron oxides (Fecarb) in Lake Xiaoxingkai compared to Lake Xingkai (P = 0.002), due to better deposition mixing, more contributions of terrigenous detrital materials, and higher OM content in Lake Xiaoxingkai. Results of structural equation modeling showed that Fecarb and total iron content (TFe) regulated 25% of DIR in Lake Xiaoxingkai and 76% in Lake Xingkai, accompanied by a negative effect of TFe on methanogenesis in Lake Xingkai. The relative abundance and diversity of Fe-reducing bacteria were significantly different between the two lakes, and showed a weak effect on sediment OM mineralization. Our findings emphasize the role of iron minerals and geochemical characterizations in regulating rates and pathways of OM mineralization, and deepen the understanding of carbon cycling in lake sediments.

Heavy metal dynamics at the sediment-water interface (SWI) has attracted plenty of attention due to their meticulous depiction for metal sorption-release processes. However, little is known about their concentration, migration and release characteristics underneath the ice, especially at the millimeter scale. Here we investigated dynamics of labile As, Cd, Cu, Mn, Pb and Zn by the diffusive gradients in thin-films (DGT) technique during ice-covered and ice-free periods in the Lake Xingkai basin. The concentrations of metals were relatively high at the SWI and ranged for As: 0.001~13.42 μg L−1, Cd: 0.01~0.45 μg L−1, Cu: 0.001~2.75 μg L−1, Mn: 5.31~2958.29 μg L−1, Pb: 0.06~1.43 μg L−1, and Zn: 2.92~112.96 μg L−1. Particularly, concentration of Mn was extremely higher than other studies, possibly due to diagenetic process. Labile concentrations of heavy metals in January were significantly lower than those in May, suggesting that elevated temperatures lead to the release of heavy metals from the sediment to the overlying water. Based on the Fick’s first law, the diffusive fluxes as a source of Mn (413.82-1163.25 mg·m-2·d-1) and As (3.53 -8.12 mg·m-2·d-1) indicated that heavy metals were released from sediments to the overlying water. While the diffusive fluxes as a sink of Zn (-1.80-(-2.36) mg·m-2·d-1) and Pb (-0.02-(-0.46) mg·m-2·d-1) to sediments. Redundancy Analysis (RDA) revealed that the main factors influencing the heavy metal migration were dissolved oxygen (45.6% of total explanation, P=0.01) in January, and water temperature (52.9%, P=0.006) in May. This study extends theoretical scope for understanding metal migration and release process, and provides valuable suggestions for lake management during the freezing period.

The Scaly-sided Merganser (SSME) is a globally endangered waterbird species, primarily inhabiting rivers. In recognition of its precarious status, it was added to the International Union for Conservation of Nature’s (IUCN) Red List of Threatened Species in 2002. The SSME is highly sensitive to ecological environmental changes driven by changes in hydrological connectivity, particularly within riverine habitats. Changbai Mountain, a high-latitude water tower in the Northeast Asia, is considered one of the most optimal breeding grounds for the SSME worldwide. Its extensive water system and dense primary forest create an environment particularly well-suited for this species. Over the past 40 years, rivers within Changbai Mountain have experienced phases of hydropower development and ecological restoration. These activities have dramatically altered the hydrological connectivity of the area. However, the response of SSME to these changes in aquatic ecology, driven by shifts in hydrological connectivity, remains uncertain. We employed the AutoRegressive Integrated Moving Average (ARIMA) model to analyze changes in the population size of the SSME. Furthermore, the GeoDetector model was used to understand the impact of ecological changes on the maximum population size (MPS) and distribution of SSME in the context of alterations in hydrological connectivity. Our findings reveal that the restoration of hydrological connectivity, a method of ecological restoration, exerts a bidirectional influence on both the distribution and the MPS of SSME. Our findings can be summarized as follows. Firstly, there was a general trend of increase in both the distribution points and population size of the SSME. Secondly, the contribution of driving factors to the MPS of SSME was ranked in the following order: temperature > Connectivity Status Index (CSI) > river width > fish abundance > river depth > velocity > altitude > forest type. Thirdly, the impacts on the distribution and MPS of SSME were not due to a single factor, but rather a complex interplay of multiple factors. Finally, the construction of small hydropower stations significantly disrupted the hydrological connectivity of rivers, thereby altering the natural habitat of the SSME. The reduction in fish population, resulting from the loss of hydrological connectivity, adversely affected the MPS. Conversely, an increase in water temperature due to loss of hydrological connectivity positively impacted the MPS. However, these effects displayed heterogeneity across different rivers. Our study suggests that the construction of gravel dams may be a suitable approach in certain sections of the river, with a recommended height limit of 1.5 m. This research provides valuable insights for managers, offering a sustainable foundation for future energy planning and ecological restoration efforts.

Hydrological connectivity is crucial for supporting aquatic biodiversity and serving ecosystem function in river floodplain wetlands. Although the effects of longitudinal connectivity in the composition of snail (Mollusca: Gastropoda) assemblages is well understood, the effects of lateral connectivity are less evaluated. Here, we evaluated the effects of lateral and longitudinal connectivity on multiple facets of alpha and beta diversity (i.e., taxonomic, functional, and phylogenetic) of freshwater snails in 38 floodplain wetlands in the Da Xing'an Mountains of Northeast China. A total of 9,784 snails subjected to 5 orders, 14 families, 24 genera, and 42 species were collected. Overall, the snail alpha diversity was higher in the nearly isolated wetlands and downstream areas. Multivariate analyses revealed that the taxonomic, functional, and phylogenetic structures of snail assemblages in floodplain wetlands differed significantly across the lateral and longitudinal gradients. Waterscape and climate variables appeared to equivalently explain a large proportion of variations in the taxonomic, functional, and phylogenetic structures of snail assemblages. Lateral connectivity and river order were paramount variables that explain the greatest variation in taxonomic, functional, and phylogenetic structures of the snail assemblages. Our study suggests that hydrological connectivity is a key factor controlling the multiple facets of snail diversity in floodplains. Attention should be paid to the effects of changing climate and waterscape on the multi-faceted diversity of snail assemblages. Due to the extant patterns in assemblage structures, floodplain restoration and management should consider the full spectrum of longitudinal and laterally connectedness to maximize aquatic biodiversity.

The Oriental rat snake Ptyas mucosa is a common non-venomous snake of the colubrid family, spanning most of South and Southeast Asia. P. mucosa is widely bred for its uses in traditional medicine, scientific research, and handicrafts. Therefore, genome resources of P. mucosa could play an important role in the efficacy of traditional medicine and the analysis of the living environment of this species. Here, we present a highly continuous P. mucosa genome with a size of 1.74 Gb. Its scaffold N50 length is 9.57 Mb, and the maximal scaffold length is 78.3 Mb. Its CG content is 37.9%, and its gene integrity reaches 86.6%. Assembled using long-reads, the total length of the repeat sequences in the genome reaches 735 Mb, and its repeat content is 42.19%. Finally, 24,869 functional genes were annotated in this genome. This study may assist in understanding P. mucosa and supporting medicinal research.

Phosphorus (P) is an essential macronutrient crucial for both plant growth and crop production, playing a pivotal role in agriculture since the early 20th century. The symbiotic relationship between AMF and plants serves as a classic illustration. These fungi play a regulatory role in the growth and development of plants, especially in facilitating the absorption of P and carbon molecules by plants. While there has been a growing body of research on the community assembly of arbuscular mycorrhizal fungi (AMF) in recent decades, our knowledge of the processes governing the coexistence of these AMF communities influenced by P in agroecosystems remains limited. To investigate the impact of various P fertilizers on AMF communities in temperate agroecosystems, this study was conducted using soils sourced from wheat–maize rotation farmland at Henan Agricultural University Yuanyang Base. With the Illumina MiSeq high-throughput sequencing technique, we systematically examined the taxonomic composition of soil AMF at the Yuanyang Base of Henan Agricultural University in a wheat–maize rotation agricultural field. Our primary objective was to unravel the mechanisms behind AMF community assembly and stability under varying P gradient fertilization conditions. Nonmetric Multidimensional Scaling (NMDS) analysis revealed significant differences among AMF communities in field soil subjected to various treatments (p < 0.05). A torus translations test demonstrated positive associations with the three treatments in 36 out of the 51 examined AMF operational taxonomic units (OTUs), making up 70.59% (p < 0.05) of the results. Furthermore, 37.84% (14/37) of the OTUs displayed preferences for the low P concentration treatment, while 34.29% (12/35) and 32.26% (10/31) favored medium and high P concentrations, respectively. An analysis of the Normalized Stochasticity Ratio (NST) and Checkerboard Score (C-score) indicated that in temperate agroecosystems, deterministic processes predominantly governed AMF in all treatment groups, with high P conditions exerting a stronger influence than low or medium P conditions. This study underscores the profound impact of long-term P fertilizer application on AMF community structures within temperate agricultural systems employing wheat–maize rotation. Additionally, it highlights the dominant role played by deterministic processes in shaping the assembly of AMF communities in these temperate agricultural systems that use P fertilizers. These findings emphasize the need for balanced nutrient management, particularly concerning P, to ensure the stability of AMF communities.

Excessive accumulation of heavy metals in global lake sediments poses a serious threat to lake water quality and ecosystem security. However, there is still a knowledge gap in comparison of heavy metal variation and pollution in hydrologically connected lakes. In this study, concentrations of As, Cd, Cr, Cu, Hg, Pb, and Zn in sediments of two hydrologically connected lakes, Xingkai Lake and Xiaoxingkai Lake, were determined during the hydrologically connected periods (May and September) and disconnected period (January and July) in 2021. We found the range of As was 2.58∼14.35 mg/kg, Cd was 0.050∼0.21 mg/kg, Cr was 28.58∼262.3 mg/kg, Cu was 3.12∼28.05 mg/kg, Hg was 0.0030∼0.14 mg/kg, Pb was 10.87∼58.86 mg/kg, and Zn was 18.21∼90.73 mg/kg. Heavy metal concentrations were lower than grade I level in Chinese soil quality standards with significant spatial and temporal differences in the basin. Overall, most of the sampling sites in Xingkai Lake and Xiaoxingkai Lake were at the uncontaminated level and moderate ecological risk during the sampling period. Two lakes showed different heavy metal compositions, accompanied by higher contamination level and higher potential ecological risk in the small lake than those in the large lake based on analysis of the geo-accumulation index and potential ecological risk index. Besides, the contamination level and potential ecological risk in May and September were higher than those in January and July, mainly due to human activities and hydrological connectivity. The ecological risks were moderate for Cd and Hg, and low for As, Cr, Cu, Pb, and Zn. Correlation and PCA analyses showed that Cd mainly originated from anthropogenic sources, while other metals mainly came from natural sources. These findings elucidate the effects of agriculture and hydrological connectivity on heavy metals in sediments, and provide scientific basis for the reasonable management of lake ecosystem.

The ice cover in winter as a physical barrier and duration would profoundly impact on changes in endogenous metal loading, migration, and transformation. Although a gradual reduction in duration and occurrence of lake ice cover in boreal lake ecosystems has been confirmed, little attention to the ice-covered period is received compared to open water studies. In this study, novel-developed diffusive gradients in thin films (DGT, ZrO-Chelex) probes were deployed to obtain the in-situ and high-resolution information on metals (Cu, Zn, Pb, Mn, Cd, Cr, and As) at the sediment-water interface (SWI) in a seasonal ice-covered lake, Chagan Lake. In addition, “source-sink” characteristics of each metal related to their endogenous release were determined based on Fick’s first law. Concentrations of labile metals at the SWI demonstrated significant spatial heterogeneity, peaking exactly below the SWI. Compared with other similar studies, concentrations of Pb (0.55 μg/L), Cr (0.58 μg/L), and As (2.4 μg/L) were a little higher even under-ice than that in other freshwater rivers and lakes, indicating potential pollution due to the agricultural intensification and petroleum extraction. The apparent diffusive fluxes suggested that sediments acted as a sink for Pb (−0.01 mg m–2 day–1), Cr (−2.37 mg m–2 day–1), and Cd (−0.1 mg m–2 day–1), diffusing from the overlying water into the sediment, while Cu (0.12 mg m–2 day–1), Zn (0.75 mg m–2 day–1), Mn (15.89 mg m–2 day–1), and As (2.12 mg m–2 day–1) as a source from sediments into the overlying water. Dissolved oxygen was the principal factor (79.5%, P = 0.032), determining the variation of the available metals at the SWI. As the urgent need for research focused on under-ice ecosystem dynamics, this study addressed the previously unknown behavior of the labile metals at the SWI and provided a unique perspective for the lake management during the ice-cover periods when external nutrient input was cut off.

Flower color is an important trait in Brassica species. However, genes responsible for the dark yellow flower trait in Chinese cabbage have not been reported. In this study, we identified a dark-yellow-flowered Chinese cabbage line SD369. Genetic analysis indicated that the dark yellow flower trait in SD369 was controlled by a single recessive locus, Br-dyp1 (dark yellow petal color 1 in Brassica rapa). Using bulked segregant RNA sequencing and kompetitive allele-specific PCR assays, Br-dyp1 was fine-mapped to an interval of 53.6 kb on chromosome A09. Functional annotation analysis, expression analysis, and sequence variation analysis revealed that Bra037130 (BraA09.ZEP), which encodes a zeaxanthin epoxidase, was the most likely candidate gene for Br-dyp1. Carotenoid profile analysis suggested that Bra037130 (BraA09.ZEP) might participate in the epoxidation from zeaxanthin to violaxanthin. The 679 bp insertion in dark yellow petal caused premature stop codon, thus caused the loss-of-function of the enzyme zeaxanthin epoxidase (ZEP), which disturbed the carotenoid metabolism, and caused the increased accumulation of total carotenoid, and finally converted the flower color from yellow to dark yellow. Comparative transcriptome analysis also showed that the “carotenoid biosynthesis” pathway was significantly enriched, and genes involved in carotenoid degradation and abscisic acid biosynthesis and metabolism were significantly downregulated. Furthermore, we developed and validated the functional marker Br-dyp1-InDel for Br-dyp1. Overall, these results provide insight into the molecular basis of carotenoid-based flower coloration in B. rapa and reveal valuable information for marker-assisted selection breeding in Chinese cabbage.

The SHPRH (SNF2, histone linker, PHD, RING, helicase) subfamily belonging to ATP-dependent chromatin remodeling factor is the effective tumor-suppressor, which can polyubiquitinate PCNA (proliferating cell nuclear antigen) and participate in post-replication repair in human. However, little is known about the functions of SHPRH proteins in plants. In this study, we identified a novel SHPRH member BrCHR39 and obtained BrCHR39-silenced transgenic Brassica rapa. In contrast to wild-type plants, transgenic Brassica plants exhibited a released apical dominance phenotype with semi-dwarfism and multiple lateral branches. Furthermore, a global alteration of DNA methylation in the main stem and bud appeared after silencing of BrCHR39. Based on the GO (gene ontology) functional annotation and KEGG (Kyoto encyclopedia of genes and genomes) pathway analysis, the plant hormone signal transduction pathway was clearly enriched. In particular, we found a significant increase in the methylation level of auxin-related genes in the stem, whereas auxin- and cytokinin-related genes were hypomethylated in the bud of transgenic plants. In addition, further qRT-PCR (quantitative real-time PCR) analysis revealed that DNA methylation level always had an opposite trend with gene expression level. Considered together, our findings indicated that suppression of BrCHR39 expression triggered the methylation divergence of hormone-related genes and subsequently affected transcription levels to regulate the apical dominance in Brassica rapa.

As microorganisms are very sensitive to changes in the lake environment, a comprehensive and systematic understanding of the structure and diversity of lake sediment microbial communities can provide feedback on sediment status and lake ecosystem protection. Xiao Xingkai Lake (XXL) and Xingkai Lake (XL) are two neighboring lakes hydrologically connected by a gate and dam, with extensive agricultural practices and other human activities existing in the surrounding area. In view of this, we selected XXL and XL as the study area and divided the area into three regions (XXLR, XXLD, and XLD) according to different hydrological conditions. We investigated the physicochemical properties of surface sediments in different regions and the structure and diversity of bacterial communities using high-throughput sequencing. The results showed that various nutrients (nitrogen, phosphorus) and carbon (DOC, LOC, TC) were significantly enriched in the XXLD region. Proteobacteria, Firmicutes, and Bacteroidetes were the dominant bacterial phyla in the sediments, accounting for more than 60% of the entire community in all regions. Non-metric multidimensional scaling analysis and analysis of similarities confirmed that β-diversity varied among different regions. In addition, the assembly of bacterial communities was dominated by a heterogeneous selection in different regions, indicating the important influence of sediment environmental factors on the community. Among these sediment properties, the partial least squares path analysis revealed that pH was the best predictor variable driving differences in bacterial communities in different regions, with higher pH reducing beta diversity among communities. Overall, our study focused on the structure and diversity of bacterial communities in lake sediments of the Xingkai Lake basin and revealed that high pH causes the β-diversity of bacterial communities in the sediment to decrease. This provides a reference for further studies on sediment microorganisms in the Xingkai Lake basin in the future.

Plasmodiophora brassicae, an obligate biotrophic pathogen-causing clubroot disease, can seriously affect Brassica crops worldwide, especially Chinese cabbage. Understanding the transcriptome and metabolome profiling changes during the infection of P. brassicae will provide key insights in understanding the defense mechanism in Brassica crops. In this study, we estimated the phytohormones using targeted metabolome assays and transcriptomic changes using RNA sequencing (RNA-seq) in the roots of resistant (BrT24) and susceptible (Y510-9) plants at 0, 3, 9, and 20 days after inoculation (DAI) with P. brassicae. Differentially expressed genes (DEGs) in resistant vs. susceptible lines across different time points were identified. The weighted gene co-expression network analysis of the DEGs revealed six pathways including “Plant–pathogen interaction” and “Plant hormone signal transduction” and 15 hub genes including pathogenic type III effector avirulence factor gene (RIN4) and auxin-responsive protein (IAA16) to be involved in plants immune response. Inhibition of Indoleacetic acid, cytokinin, jasmonate acid, and salicylic acid contents and changes in related gene expression in R-line may play important roles in regulation of clubroot resistance (CR). Based on the combined metabolome profiling and hormone-related transcriptomic responses, we propose a general model of hormone-mediated defense mechanism. This study definitely enhances our current understanding and paves the way for improving CR in Brassica rapa.

Clubroot, caused by the soil-borne protist Plasmodiophora brassicae, is one of the most destructive diseases of Chinese cabbage worldwide. However, the clubroot resistance mechanisms remain unclear. In this study, in both clubroot-resistant (DH40R) and clubroot-susceptible (DH199S) Chinese cabbage lines, the primary (root hair infection) and secondary (cortical infection) infection stages started 2 and 5 days after inoculation (dai), respectively. With the extension of the infection time, cortical infection was blocked and complete P. brassica resistance was observed in DH40R, while disease scales of 1, 2, and 3 were observed at 8, 13, and 22 dai in DH199S. Transcriptome analysis at 0, 2, 5, 8, 13, and 22 dai identified 5,750 relative DEGs (rDEGs) between DH40R and DH199S. The results indicated that genes associated with auxin, PR, disease resistance proteins, oxidative stress, and WRKY and MYB transcription factors were involved in clubroot resistance regulation. In addition, weighted gene coexpression network analysis (WGCNA) identified three of the modules whose functions were highly associated with clubroot-resistant, including ten hub genes related to clubroot resistance (ARF2, EDR1, LOX4, NHL3, NHL13, NAC29, two AOP1, EARLI 1, and POD56). These results provide valuable information for better understanding the molecular regulatory mechanism of Chinese cabbage clubroot resistance.

The global increase in occurrence of harmful algae blooms in lakes has gained widespread attention. Although N and P are the main factors for primary productivity in lake ecosystems, dissolved iron (Fe) plays a crucial role as an essential micronutrient for phytoplankton growth. The interaction between Fe and light in phytoplankton growth is reasonably well studied in marine ecosystems, but there is a lack of understanding of this interaction in lakes. We conducted both a field study and an incubation experiment to identify the role of Fe and light in modulating phytoplankton growth and species composition in a eutrophic lake, Lake Xingkai, Northeast China. We found that the interactions between Fe bioavailability and light intensity impact phytoplankton community structure and Fe uptake. Phytoplankton composition and abundance varied greatly in response to elevated light and Fe. Bacillariophyta are favored under light-Fe co-limitation, while the dominant taxa changed from Bacillariophyta to Cyanophyta and Chlorophyta under elevated Fe and light conditions. We unveil the competitive advantage of a common bloom-forming and N2-fixing algae Anabaena azotica relative to other species under high light and high Fe conditions. Our findings expand the knowledge of Fe-light interactions on phytoplankton growth and provide fresh insight into phytoplankton community responses to variation in light and Fe in eutrophic lakes. This information is important for efficient eutrophication control and lake management.

Along with being important pigments that determining the flower color in many plants, anthocyanins also perform crucial functions that attract pollinators and reduce abiotic stresses. Purple and white are two different colors of radish petals. In this study, two cDNA libraries constructed with purple and white petal plants were sequenced for transcriptome profiling. Transcriptome results implied that the expression level of the genes participating in the anthocyanin biosynthetic pathway was commonly higher in the purple petals than that in the white petals. In particular, two genes, F3′H and DFR, had a significantly higher expression pattern in the purple petals, suggesting the important roles these genes playing in radish petal coloration. BSA-seq aided-Next Generation Sequencing of two DNA pools revealed that the radish purple petal gene (RsPP) was located on chromosome 7. With additional genotyping of 617 F2 population plants, the RsPP was further confined within a region of 93.23 kb. Transcriptome and Sanger sequencing analysis further helped identify the target gene, Rs392880. Rs392880 is a homologous gene to F3′H, a key gene in the anthocyanin biosynthetic pathway. These results will aid in elucidating the molecular mechanism of plant petal coloration and developing strategies to modify flower color through genetic transformation.

Flower color is an important trait in plants. However, genes responsible for the white flower trait in Chinese cabbage are rarely reported. In this study, we constructed an F2 population derived from the Y640-288 (white flower) and Y641-87 (yellow flower) lines for the fine mapping of the white flower gene BrWF3 in Chinese cabbage. Genetic analysis indicated that BrWF3 was controlled by a single recessive gene. Using BSA-seq and KASP assays, BrWF3 was fine-mapped to an interval of 105.6 kb. Functional annotation, expression profiling, and sequence variation analyses confirmed that the AtPES2 homolog, Bra032957, was the most likely candidate gene for BrWF3. Carotenoid profiles and transmission electron microscopy analysis suggested that BrWF3 might participate in the production of xanthophyll esters (particularly violaxanthin esters), which in turn disrupt chromoplast development and the formation of plastoglobules (PGs). A SNP deletion in the third exon of BrWF3 caused the loss of protein function, and interfered with the normal assembly of PGs, which was associated with reduced expression levels of genes involved in carotenoid metabolism. Furthermore, we developed and validated the functional marker TXBH83 for BrWF3. Our results provide insight into the molecular mechanism underlying flower color pigmentation and reveal valuable information for marker-assisted selection (MAS) breeding in Chinese cabbage.

Chinese cabbage is one of the most important and widely consumed vegetables in China. The developmental transition from the vegetative to reproductive phase is a crucial process in the life cycle of flowering plants. In spring-sown Chinese cabbage, late bolting is desirable over early bolting. In this study, we analyzed double haploid (DH) lines of late bolting (“Y410-1” and “SY2004”) heading Chinese cabbage (Brassica rapa var. pekinensis) and early-bolting Chinese cabbage (“CX14-1”) (B. rapa ssp. chinensis var. parachinensis) by comparative transcriptome profiling using the Illumina RNA-seq platform. We assembled 721.49 million clean high-quality paired-end reads into 47,363 transcripts and 47,363 genes, including 3,144 novel unigenes. There were 12,932, 4,732, and 4,732 differentially expressed genes (DEGs) in pairwise comparisons of Y410-1 vs. CX14-1, SY2004 vs. CX14-1, and Y410-1 vs. SY2004, respectively. The RNA-seq results were confirmed by reverse transcription quantitative real-time PCR (RT-qPCR). A Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of DEGs revealed significant enrichment for plant hormone and signal transduction as well as starch and sucrose metabolism pathways. Among DEGs related to plant hormone and signal transduction, six unigenes encoding the indole-3-acetic acid-induced protein ARG7 (BraA02g009130), auxin-responsive protein SAUR41 (BraA09g058230), serine/threonine-protein kinase BSK11 (BraA07g032960), auxin-induced protein 15A (BraA10g019860), and abscisic acid receptor PYR1 (BraA08g012630 and BraA01g009450), were upregulated in both late bolting Chinese cabbage lines (Y410-1 and SY2004) and were identified as putative candidates for the trait. These results improve our understanding of the molecular mechanisms underlying flowering in Chinese cabbage and provide a foundation for studies of this key trait in related species.

Clubroot disease caused by Plasmodiophora brassicae is one of the major threats to Brassica crops. New clubroot resistant varieties of Chinese cabbage (B. rapa ssp. pekinensis) have been developed through breeding, but the underlying genetic mechanism of clubroot resistance is still unclear. In this study, two Chinese cabbage DH lines, clubroot-resistant Y635-10 and susceptible Y177-47 were crossed to develop F2 population for fine mapping and cloning resistance gene CRq. After sequence analysis, the expression vector was constructed by gateway technology and transferred into Arabidopsis thaliana for functional characterization. Bulked segregant analysis sequencing (BSA-seq) confirmed that CRq is located in the 80 kb genomic region on chromosome A03 between markers GC30-FW/RV and BGA. In silico tools confirmed that the gene length was 3959 bp with 3675 bp coding sequences (CDs), and it has three exons and two introns. In addition, we found 72bp insertion in the third exon of CRq in the susceptible line. We developed and verified functional marker Br-insert1, by which genotyping results showed that 72bp insertion might lead to the destruction of the LRR region of Y177-47, resulting in a loss of resistance relative to clubroot. The results of genetic transformation showed that the roots for wild-type Arabidopsis thaliana were significantly enlarged compared with T2 generation transgenic Arabidopsis after treatment by P. brassicae spores, and transgenic Arabidopsis had certain resistance. Therefore, CRq is a candidate gene of clubroot disease resistance in Chinese cabbage, which could be used as a reference for elucidating disease resistance mechanisms and the marker-assisted breeding of clubroot resistant varieties.

Self-incompatibility (SI) is a genetic mechanism flowering plants adopted to reject self-pollen and promote outcrossing. In the Brassicaceae family plants, the stigma tissue plays a key role in self-pollen recognition and rejection. We reported earlier in Chinese cabbage (Brassica rapa) that stigma tissue showed upregulated ethylene responses and programmed cell death (PCD) upon compatible pollination, but not in SI responses. Here, we show that SI is significantly compromised or completely lost in senescent flowers and young flowers of senescent plants. Senescence upregulates senescence-associated genes in B. rapa. Suppressing their expression in young stigmas by antisense oligodeoxyribonucleotide abolishes compatible pollination-triggered PCD and inhibits the growth of compatible pollen tubes. Furthermore, ethylene biosynthesis genes and response genes are upregulated in senescent stigmas, and increasing the level of ethylene or inhibiting its response increases or decreases the expression of senescence-associated genes, respectively. Our results show that senescence causes PCD in stigmatic papilla cells and is associated with the breakdown of SI in Chinese cabbage and in radish.

As an important appearance trait, the rind color of watermelon fruit affects the commodity value and further determines consumption choices. In this study, a comparative transcriptome analysis was conducted to elucidate the genes and pathways involved in the formation of yellow rind fruit in watermelon using a yellow rind inbred line WT4 and a green rind inbred line WM102. A total of 2,362 differentially expressed genes (DEGs) between WT4 and WM102 at three different stages (0, 7, and 14 DAP) were identified and 9,770 DEGs were obtained by comparing the expression level at 7 DAP and 14 DAP with the former stages of WT4. The function enrichment of DEGs revealed a number of pathways and terms in biological processes, cellular components, and molecular functions that were related to plant pigment metabolism, suggesting that there may be a group of common core genes regulating rind color formation. In addition, next-generation sequencing aided bulked-segregant analysis (BSA-seq) of the yellow rind pool and green rind pool selected from an F2 population revealed that the yellow rind gene (Clyr) was mapped on the top end of chromosome 4. Based on the BSA-seq analysis result, Clyr was further confined to a region of 91.42 kb by linkage analysis using 1,106 F2 plants. These results will aid in identifying the key genes and pathways associated with yellow rind formation and elucidating the molecular mechanism of rind color formation in watermelon.

Chinese cabbage (Brassica rapa L.) is one of the most important and highly nutritious vegetables in China belonging to the Brassicaceae family. Flowering or bolting is one of the most critical developmental stages in flowering plants. For the spring-sown Chinese cabbage, late-bolting is desirable over early-bolting according to consumer preferences. We determined the inheritance pattern of the late-bolting trait using F1 and F2 generated from a cross between ‘SY2004’ (late-bolting) and ‘CX14-1’ (early-bolting). The genetic analysis revealed that the late-bolting to early-bolting trait was controlled by an incomplete dominant gene that we named BrLb-1. Furthermore, we performed bulked segregant analysis (BSA) via whole genome re-sequencing and the results showed that this gene was harbored on the chromosome A07 at the intersections of 20,070,000 to 25,290,000 bp and 20,330,000 to 25,220,000, an interval distance of 4.89 Mb. In this candidate interval, totals of 2321 and 1526 SNPs with non-synonymous mutations, and 229 and 131 InDels with frameshift mutations, were found between the parents and the bulked pools, respectively. Furthermore, we identified three putative candidate genes for the late-bolting trait, including BraA07g029500, BraA07g029530 and BraA07g030360, which code for the AGAMOUS-like MADS-box protein AGL12, a pentatricopeptide repeat-containing protein and NAC transcription factor 29, respectively; however, further functional analysis is required. These genetic variants could be utilized for the further development of molecular markers for marker-assisted breeding in Chinese cabbage.

Kompetitive allele-specific PCR (KASP) is a cost-effective single-step SNP genotyping technology, With an objective to enhance the marker repertoire and develop high efficient KASP-SNP markers in Chinese cabbage, we re-sequenced four Chinese cabbage doubled haploid (DH) lines, Y177-47, Y635-10, Y510-1 and Y510-9, and generated a total of more than 38.5 billion clean base pairs. A total of 827,720 SNP loci were identified with an estimated density of 3,217 SNPs/Mb. Further, a total of 387,354 SNPs with at least 30 bp to the next most adjacent SNPs on either side were selected as resource for KASP markers. From this resource, 258 (96.27%) of 268 SNP loci were successfully transformed into KASP-SNP markers using a Roche LightCycler 480-II instrument. Among these markers, 221 (85.66%) were co-dominant markers, 220 (85.27%) were non-synonymous SNPs, and 257 (99.6%) were newly developed markers. In addition, 53 markers were applied for genotyping of 34 Brassica rapa accessions. Cluster analysis separated these 34 accessions into three clusters based on heading types. The millions of SNP loci, a large set of resource for KASP markers, as well as the newly developed KASP markers in this study may facilitate further genetic and molecular breeding studies in Brassica rapa.

Ogura cytoplasmic male sterility (Ogura CMS), originally identified in wild radish (Raphanus sativus), has enabled complete pollen sterility in Brassica plants, but the underlying mechanism in Ogura CMS Chinese cabbage (Brassica rapa ssp. pekinensis) remains unclear. In this study cytological analysis showed that during microsporogenesis the meiosis occurred normally, and the uninucleated pollens subsequently formed, but the development of both binucleated and trinucleated pollens was obviously disrupted due to defects of pollen mitosis in the Ogura CMS line (Tyms) compared with the corresponding maintainer line (231–330). In transcriptome profiling a total of 8052 differentially expressed genes (DEGs) were identified, among which 3890 were up-regulated and 4162 were down-regulated at the pollen abortion stages in an Ogura CMS line. KOG cluster analysis demonstrated that a large number of DEGs were related to the cytoskeleton’s dynamics, which may account for the failure of pollen mitosis during development in the Ogura CMS line. The pivotal genes related to the phenylpropane synthesis pathway (PAL, 4CL and CAD) were significantly down-regulated, which probably affected the formation and disposition of anther lignin and sporopollenin, and eventually led to abnormality in the pollen exine structure. In addition, several key up-regulated genes (GPX7, G6PD and PGD1) related to the glutathione oxidation-reduction (REDOX) reaction indicated that the accumulation of peroxides in Ogura CMS lines during this period affected the pollen development. Taken together, this cytological and molecular evidence is expected to advance our understanding of pollen abortion induced by Ogura cytoplasmic action in Chinese cabbage.

Raphanus sativus, an important cruciferous vegetable, has been increasingly affected by clubroot disease. Establishing a stable and accurate resistance identification method for screening resistant germplasms is urgently needed in radish. In this study, the influence of inoculum concentration, inoculation methods, and pH of the substrate on disease occurrence was studied. The result showed that the disease index (DI) was highest at 2 × 108 spores/mL, the efficiency of two-stage combined inoculation methods was higher than others, and pH 6.5 was favorable for the infection of P. brassicae. By using this new method, DIs of 349 radish germplasms varying from 0.00 to 97.04, presented significantly different levels of resistance. Analysis showed that 85.06% germplasms from China were susceptible to P. brassicae, whilst 28 accessions were resistant and mainly distributed in east, southwest, northwest, and south-central China. Most of the exotic germplasms were resistant. Repeated experiments verified the stability and reliability of the method and the identity of germplasm resistance. In total, 13 immune, 5 highly resistant and 21 resistant radish accessions were identified. This study provides an original clubroot-tolerance evaluation technology and valuable materials for the development of broad-spectrum resistant varieties for sustainable clubroot management in radish and other cruciferous crops.

Increasing evidence has revealed that microRNAs play a pivotal role in the post transcriptional regulation of gene expression in response to pathogens in plants. However, there is little information available about the expression patterns of miRNAs and their targets in Chinese cabbage (Brassica rapa ssp. pekinensis) under Plasmodiophora brassicae stress. In the present study, using deep sequencing and degradome analysis, a genome-wide identification of miRNAs and their targets during P. brassicae stress was performed. A total of 221 known and 93 potentially novel miRNAs were successfully identified from two root libraries of one control (635-10CK) and P. brassicae-treated Chinese cabbage samples (635-10T). Of these, 14 known and 10 potentially novel miRNAs were found to be differentially expressed after P. brassicae treatment. Degradome analysis revealed that the 223 target genes of the 75 miRNAs could be potentially cleaved. KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis suggested that the putative target genes of the miRNAs were predominately involved in selenocompound metabolism and plant hormone signal transduction. Then the expression of 12 miRNAs was validated by quantitative real-time PCR (qRT-PCR). These results provide insights into the miRNA-mediated regulatory networks underlying the stress response to the plant pathogen P. brassicae.