Quantitative proteomics analysis identified new interacting proteins of JAL30 in Arabidopsis.

Jacalin-related lectins (JALs) are a unique group of plant lectins derived from the jacalin protein family, which play important roles in plant defense responses. JAL30/PBP1 (PYK10 binding protein 1) interacts with inactive PYK10, exerting negative regulatory control over the size of the PYK10 complex, which is formed and activated upon insect or pathogen invasion. However, the precise interplay between JAL30 and other components remains elusive. In this study, we found JAL30 as a nucleocytoplasmic protein, but no obvious phenotype was observed in jal30-1 single mutant. Through immunoprecipitation (IP) enrichment combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS), dozens of new JAL30 interacting proteins were found in addition to several reported ones. Gene Ontology (GO) analysis revealed that these interacting proteins were highly related to the wounding and bacterial stimuli, suggesting their potential involvement in the jasmonate (JA) response. Importantly, the expression of JAL30 was induced by MeJA treatment, further highlighting its relevance in plant defense mechanisms. A novel JAL30 interacting protein, ESM1, was identified and its interaction with JAL30 was confirmed by Co-immunoprecipitation. Moreover, ESM1 was found as an O-GlcNAcylated protein, suggesting that JAL30 may possess glycosylated protein binding ability, particularly in O-GlcNAcylated protein and peptide recognition. Overall, our study provides valuable insights into the interacting protein network and biological function of JAL30, demonstrates the interaction between JAL30 and ESM1, and uncovers the potential significance of JAL30 in plant defense system, potentially through its association with PYK10 complex or JA response. SIGNIFICANCE: The biological functions of lectin proteins, including defense responses, immunity responses, signal transduction, have been well studied. Lectin proteins were also utilized to enrich glycosylated proteins for their specific carbohydrates binding capability. Jacalin-related lectins (JALs) were found to involve in plant defense mechanism. However, it is not yet clear whether JALs could use for enrichment of glycosylated proteins. In this study, we used label-free quantification method to identify interacting proteins of JAL30. A novel interacting protein, ESM1, as an O-GlcNAcylated protein was found. ESM1 has been reported to take part in defense against insect herbivory. Therefore, our findings provided experimental evidence to confirm that JALs have potential to be developed as the bio-tools to enrich glycosylated proteins. Finally, our data not only illustrated the vital biological role of JALs in plants, but also verified unique function of JAL30 in recognizing O-GlcNAcylated proteins.

Fungal Community Succession and Volatile Compound Changes during Fermentation of Laobaigan Baijiu from Chinese Hengshui Region.

To investigate the core fungal community succession and its effects of volatile compound production during different stages (D-1, D-2, D-3, E-4, E-5, and E-6) of Hengshui Laobaigan Baijiu, high-throughput sequencing (HTS) was carried out, accompanied by the identification and quantification of the volatile flavor compounds using headspace solid-phase coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS). HTS results demonstrated that the fungal community of stage D-1 was similar to that of E-4 after adding Daqu, while the richness and diversity of the fungal community were most prominent at stage E-6. Moreover, the addition of Daqu at the beginning of Ercha fermentation resulted in a significant increase in the relative abundances of the fungal community at the genus level, setting the stage for the production of volatile compounds. GC-MS analysis revealed the presence of a total of 45 volatile compounds. Combining the GC-MS result with the heat map and principal component analysis (PCA), the concentrations of volatile compounds were highest in stage E-5. Meanwhile, concentrations of esters, especially ethyl acetate, ethyl lactate, isoamyl acetate and ethyl hexanoate, were high in both stages E-5 and E-6. This indicated that stage E-5 was crucial to the fermentation process of Laobaigan Baijiu. Three fungal genera (Saccharomyces, Candida, and Pichia) were indicated as the core microbiota for the production of the main volatile flavor compounds of Laobaigan Baijiu through partial least square (PLS) analysis. The information provided in this study offered valuable insights into the fermentation mechanism of Laobaigan Baijiu, thereby serving as a theoretical framework for enhancing the quality of Baijiu and realizing cost-effective production.

Identification of novel molecular subtypes to improve the classification framework of nasopharyngeal carcinoma.

BACKGROUND: Nasopharyngeal carcinoma (NPC) treatment is largely based on a 'one-drug-fits-all' strategy in patients with similar pathological characteristics. However, given its biological heterogeneity, patients at the same clinical stage or similar therapies exhibit significant clinical differences. Thus, novel molecular subgroups based on these characteristics may better therapeutic outcomes. METHODS: Herein, 192 treatment-naïve NPC samples with corresponding clinicopathological information were obtained from Fujian Cancer Hospital between January 2015 and January 2018. The gene expression profiles of the samples were obtained by RNA sequencing. Molecular subtypes were identified by consensus clustering. External NPC cohorts were used as the validation sets. RESULTS: Patients with NPC were classified into immune, metabolic, and proliferative molecular subtypes with distinct clinical features. Additionally, this classification was repeatable and predictable as validated by the external NPC cohorts. Metabolomics has shown that arachidonic acid metabolites were associated with NPC malignancy. We also identified several key genes in each subtype using a weighted correlation network analysis. Furthermore, a prognostic risk model based on these key genes was developed and was significantly associated with disease-free survival (hazard ratio, 1.11; 95% CI, 1.07-1.16; P < 0.0001), which was further validated by an external NPC cohort (hazard ratio, 7.71; 95% CI, 1.39-42.73; P < 0.0001). Moreover, the 1-, 3-, and 5-year areas under the curve were 0.84 (95% CI, 0.74-0.94), 0.81 (95% CI, 0.73-0.89), and 0.82 (95% CI, 0.73-0.90), respectively, demonstrating a high predictive value. CONCLUSIONS: Overall, we defined a novel classification of nasopharyngeal carcinoma (immune, metabolism, and proliferation subtypes). Among these subtypes, metabolism and proliferation subtypes were associated with advanced stage and poor prognosis of NPC patients, whereas the immune subtype was linked to early stage and favorable prognosis.

Mefloquine improves pulmonary fibrosis by inhibiting the KCNH2/Jak2/Stat3 signaling pathway in macrophages.

Idiopathic pulmonary fibrosis (IPF) is a life-threatening disease characterized by severe pulmonary fibrosis, for which there is an urgent need for effective therapeutic agents. Mefloquine (Mef) is a quinoline compound primarily used for the treatment of malaria. However, high doses (>25 mg/kg) may lead to side effects such as cardiotoxicity and psychiatric disorders. Here, we found that low-dose Mef (5 mg/kg) can safely and effectively treat IPF mice. Functionally, Mef can improve the pulmonary function of IPF mice (PIF, PEF, EF50, VT, MV, PENH), alleviating pulmonary inflammation and fibrosis by inhibiting macrophage activity. Mechanically, Mef probably regulates the Jak2/Stat3 signaling pathway by binding to the 492HIS site of Potassium voltage-gated channel subfamily H member 2 (KCNH2) protein in macrophages, inhibiting the secretion of macrophage inflammatory and fibrotic factors. In summary, Mef may inhibit macrophage activity by binding to KCNH2 protein, thereby slowing down the progress of IPF.

Celastrol as a candidate drug for silicosis: From bioinformatics and network pharmacology to experimental validation.

Silicosis, a highly lethal occupational respiratory disease characterized by irreversible pulmonary fibrosis, remains challenging to treat due to its unclear pathogenesis. In this study, bioinformatics, network pharmacology, and experimental validation were combined to explore potential mechanisms and therapeutic drugs for silicosis. First, the differentially expressed genes（DEGs）and pathway enrichment in pulmonary fibrosis were identified by GO and KEGG analysis. Next, the differential genes were submitted to cMap database for drug prediction and celastrol stood out as the most promising candidate drug. Then, network pharmacology analysis identified pharmacological targets of celastrol and demonstrated that celastrol could regulate JAK-STAT, MAPK, and Toll-like receptor signaling pathways. Finally, we verified the therapeutic role and mechanism of celastrol on silicosis. In vivo, celastrol significantly ameliorated CS-induced inflammation and fibrosis in silicosis mice, including inflammatory cell infiltration, collagen fiber and extracellular matrix deposition, fibroblast activation and related factor expression. Moreover, it dramatically improved lung respiratory function of silicosis mice. In vitro, celastrol suppressed CS-induced cytokine expression, apoptosis of macrophages and activation of Stat3 and Erk1/2 signals. Overall, our research identified and verified celastrol as a novel and promising candidate drug for silicosis.

Global Prevalence and Hemagglutinin Evolution of H7N9 Avian Influenza Viruses from 2013 to 2022.

H7N9 avian influenza viruses have caused severe harm to the global aquaculture industry and human health. For further understanding of the characteristics of prevalence and hemagglutinin evolution of H7N9 avian influenza viruses, we generated the global epidemic map of H7N9 viruses from 2013 to 2022, constructed a phylogenetic tree, predicted the glycosylation sites and compared the selection pressure of the hemagglutinin. The results showed that although H7N9 avian influenza appeared sporadically in other regions worldwide, China had concentrated outbreaks from 2013 to 2017. The hemagglutinin genes were classified into six distinct lineages: A, B, C, D, E and F. After 2019, H7N9 viruses from the lineages B, E and F persisted, with the lineage B being the dominant. The hemagglutinin of highly pathogenic viruses in the B lineage has an additional predicted glycosylation site, which may account for their persistent pandemic, and is under more positive selection pressure. The most recent ancestor of the H7N9 avian influenza viruses originated in September 1991. The continuous evolution of hemagglutinin has led to an increase in virus pathogenicity in both poultry and humans, and sustained human-to-human transmission. This study provides a theoretical basis for better prediction and control of H7N9 avian influenza.

Characterization of Volatile Flavor Compounds in Dry-Rendered Beef Fat by Different Solvent-Assisted Flavor Evaporation (SAFE) Combined with GC-MS, GC-O, and OAV.

To comprehensively understand the volatile flavor composition of dry-rendered beef fat, solvent-assisted flavor evaporation (SAFE) with four extraction solvents (dichloromethane, pentane, ethyl ether, and methanol) combined with gas chromatography-mass spectrometry (GC-MS) and gas chromatography-olfactormetry (GC-O) were performed. GC-MS analysis found 96 different volatile compounds in total using the four extraction solvents. According to the GC-MS results and the heat map and principal component analysis (PCA), most of the volatile compounds resulted from dichloromethane and pentane extraction, followed by ethyl ether. Methanol extraction found a few volatile compounds of higher polarity, which was supplementary to the analysis results. Moreover, GC-O analysis found 73 odor-active compounds in total using the four extraction solvents. The GC-O results found that pentane and dichloromethane extraction had a significantly larger number of odor-active compounds than ethyl ether and methanol extraction. This indicated that pentane and dichloromethane were more effective solvents for the extraction of odor-active compounds than the other two solvents. Finally, a total of 15 compounds of odor-active values (OAVs) ≥ 1 were determined to be the key aroma compounds in the dry-rendered beef fat, including 2-methyl-3-furanthiol, 3-methylthiopropanal, (E,E)-2,4-nonadienal, 12-methyltridecanal, and 1-octen-3-one.

Recent Advances in Antitumor Dendritic Cell Vaccines.

Background: Dendritic cells (DCs) are the most important antigen-presenting cells in the body and play a key role in antigen recognition, uptake, processing, and presentation and mediate nonspecific immunity and specific immunity. Purpose: To summarize the main findings that DC vaccines are a new immunotherapy scheme combining the strengths of tumor antigens and DCs that can boost the body's identification and clearance of tumors. Methods: In this review, the authors focus on the biological characteristics of DCs, recent advances in the understanding of antitumor mechanisms, and the classification of DC vaccines. Results: The current progress of DC-based vaccine immunotherapy for common tumors with high morbidity or mortality in China were systematically summarize. Conclusions: The DC vaccines combining the strengths of tumor antigens will provide directions to explore reasonable, safe, and effective combination immunotherapy strategies for tumors in the future.

[Chloroplast genomic characterization and phylogenetic analysis of Pellionia scabra].

Pellionia scabra belongs to the genus Pellionia in the family of Urticaceae, and is a high-quality wild vegetables with high nutritional value. In this study, high-throughput techniques were used to sequence, assemble and annotate the chloroplast genome. We also analyzed its structure, and construct the phylogenetic trees from the P. scabra to further study the chloroplast genome characteristics. The results showed that the chloroplast genome size was 153 220 bp, and the GC content was 36.4%, which belonged to the typical tetrad structure in P. scabra. The chloroplast genome encodes 130 genes, including 85 protein-coding genes, 37 tRNA genes, and 8 rRNA genes in P. scabra. Among them, 15 genes contained 1 intron, 2 genes contained 2 introns, and rps12 had trans-splicing, respectively. In P. scabra, chloroplast genomes could be divided into four categories, including 43 photosynthesis, 64 self-replication, other 7 coding proteins, and 4 unknown functions. A total of 51 073 codons were detected in the chloroplast genome, among which the codon encoding leucine (Leu) accounted for the largest proportion, and the codon preferred to use A and U bases. There were 72 simple sequence repeats (SSRs) in the chloroplast genome of P. scabra, containing 58 single nucleotides, 12 dinucleotides, 1 trinucleotide, and 1 tetranucleotide. The ycf1 gene expansion was present at the IRb/SSC boundary. The phylogenetic trees showed that P. scabra (OL800583) was most closely related to Elatostema stewardii (MZ292972), Elatostema dissectum (MK227819) and Elatostema laevissimum var. laevissimum (MN189961). Taken together, our results provide worthwhile information for understanding the identification, genetic evolution, and genomics research of P. scabra species.

[Characteristics and phylogenetic analysis of chloroplast genome of a new type of fruit Rubus rosaefolius].

The genomic DNA of Rubus rosaefolius was extracted and sequenced by Illumina NovaSeq platform to obtain the complete chloroplast genome sequence, and the sequence characteristics and phylogenetic analysis of chloroplast genes were carried out. The results showed that the complete chloroplast genome of the R. rosaefolius was 155 650 bp in length and had a typical tetrad structure, including two reverse repeats (25 748 bp each), a large copy region (85 443 bp) and a small copy region (18 711 bp). A total of 131 genes were identified in the whole genome of R. rosaefolius chloroplast, including 86 protein coding genes, 37 tRNA genes and 8 rRNA genes. The GC content of the whole genome was 36.9%. The genome of R. rosaefolius chloroplast contains 47 scattered repeats and 72 simple sequence repeating (SSR) loci. The codon preference is leucine codon, and the codon at the end of A/U is preferred. Phylogenetic analysis showed that R. rosaefolius had the closest relationship with R. taiwanicola, followed by R. rubraangustifolius and R. glandulosopunctatus. The chloroplast genome characteristics and phylogenetic analysis of R. rosaefolius provide a theoretical basis for its genetic diversity research and chloroplast development and utilization.

Identification and Characterization of PRE Genes in Moso Bamboo (Phyllostachys edulis).

Basic helix-loop-helix (bHLH)/HLH transcription factors are involved in various aspects of the growth and development of plants. Here, we identified four HLH genes, PePRE1-4, in moso bamboo plants that are homologous to Arabidopsis PRE genes. In bamboo seedlings, PePRE1/3 were found to be highly expressed in the internode and lamina joint by using quantitative RT-PCR analysis. In the elongating internode of bamboo shoots, PePRE genes are expressed at higher levels in the basal segment than in the mature top segment. Overexpression of PePREs (PePREs-OX) in Arabidopsis showed longer petioles and hypocotyls, as well as earlier flowering. PePRE1 overexpression restored the phenotype due to the deficiency of AtPRE genes caused by artificial micro-RNA. PePRE1-OX plants showed hypersensitivity to propiconazole treatment compared with the wild type. In addition, PePRE1/3 but not PePRE2/4 proteins accumulated as punctate structures in the cytosol, which was disrupted by the vesicle recycling inhibitor brefeldin A (BFA). PePRE genes have a positive function in the internode elongation of moso bamboo shoots, and overexpression of PePREs genes promotes flowering and growth in Arabidopsis. Our findings provided new insights about the fast-growing mechanism of bamboo shoots and the application of PRE genes from bamboo.

Effects of burdock inulin-type fructans exposure on the physiological function of healthy mice and their filial generation.

Inulin-type fructans (ITFs) have been shown to possess various biological activities. However, studies on their safety and side effects are limited. Hence, we aimed to evaluate the possible effects of burdock ITFs on the physiological indices of healthy mice and their filial generation when fed for six months. Thirty-two C57BL/6J mice were randomly divided into two groups; a normal control (NC) and an ITFs group. The parental generations were kept in one cage with free access to a normal diet and double-distilled water (P-NC group) or burdock ITFs drinking water (P-ITFs group, 2% w/v). The filial generations (F-NC group and F-ITFs group) were kept separately and were fed as their parental generation. Behavior, organ/body weight, serum indices, histopathology, time of production, and number of pup births were observed. There were no significant adverse effects on these indices. Functional indices of the spleen, lung, heart, and pancreas of the ITFs groups were higher than those of the NC groups, respectively. Interestingly, the serum glucose (GLU), total cholesterol (TC), uric acid (UA) and creatine kinase (CK) levels of the ITFs groups were lower than those of the NC groups. Meanwhile, the pregnancy number and pup birth number of the P-ITFs group were more than those of P-NC group. Therefore, long-term consumption of burdock ITFs has no obvious adverse effects on the health of parental mice and their offspring, but may contribute to reproductive capacity, fatigue reduction, and risk reduction of renal disease.

Segmentation of retinal vessels based on MRANet.

The segmentation of retinal vessel takes a crucial part in computer-aided diagnosis of diseases and eye disorders. However, the insufficient segmentation of the capillary vessels and weak anti-noise interference ability make such task more difficult. To solve this problem, we proposed a multi-scale residual attention network (MRANet) which is based on U-Net network. Firstly, to collect useful information about the blood vessels more effectively, we proposed a multi-level feature fusion block (MLF block). Then, different weights of each fused feature are learned by using attention blocks, which can retain more useful feature information while reducing the interference of redundant features. Thirdly, multi-scale residual connection block (MSR block) is constructed, which can better extract the image features. Finally, we use the DropBlock layer in the network to reduce the network parameters and alleviate network overfitting. Experiments show that based on DRIVE, the accuracy rate and the AUC performance value of our network are 0.9698 and 0.9899 respectively, and based on CHASE_DB1 dataset, they are 0.9755 and 0.9893 respectively. Our network has a better segmentation effect compared with other methods, which can ensure the continuity and completeness of blood vessel segmentation.

Characterization of Highbush Blueberry (Vaccinium corymbosum L.) Anthocyanin Biosynthesis Related MYBs and Functional Analysis of VcMYB Gene.

As one of the most important transcription factors regulating plant anthocyanin biosynthesis, MYB has attracted great attentions. In this study, we identified fifteen candidate anthocyanin biosynthesis related MYB (ABRM) proteins, including twelve R2R3-MYBs and three 1R-MYBs, from highbush blueberry. The subcellular localization prediction results showed that, with the exception of VcRVE8 (localized in chloroplast and nucleus), all of the blueberry ABRMs were nucleus-localized. The gene structure analysis revealed that the exon numbers of the blueberry ABRM genes varied greatly, ranging between one and eight. There are many light-responsive, phytohormone-responsive, abiotic stress-responsive and plant growth and development related cis-acting elements in the promoters of the blueberry ABRM genes. It is noteworthy that almost all of their promoters contain light-, ABA- and MeJA-responsive elements, which is consistent with the well-established results that anthocyanin accumulation and the expression of MYBs are influenced significantly by many factors, such as light, ABA and JA. The gene expression analysis revealed that VcMYB, VcMYB6, VcMYB23, VcMYBL2 and VcPH4 are expressed abundantly in blueberry fruits, and VcMYB is expressed the highest in the red, purple and blue fruits among all blueberry ABRMs. VcMYB shared high similarity with functionally proven ABRMs from many other plant species. The gene cloning results showed that VcMYB had three variable transcripts, but only the transient overexpression of VcMYB-1 promoted anthocyanin accumulation in the green fruits. Our study can provide a basis for future research on the anthocyanin biosynthesis related MYBs in blueberry.

Novel nitrogen mustard-artemisinin hybrids with potent anti-leukemia action through DNA damage and activation of GPx.

The incidence of various types of cancers is increasing every year. Among these, leukemia is extremely common, and thus, developing novel drugs to combat leukemia is crucial. In this study, we designed and synthesized several hybrids and obtained a new lead molecule 5a, with a strong therapeutic effect on leukemia. The results indicated that most hybrids effectively inhibited the growth of leukemia cells, HCT-116, and A549 cancer cells with an IC50 of <10 µM. Among these hybrids, 5a and 4h showed significant anticancer activity against CCRF-CEM, with IC50 values of 0.895 µM and 0.555 µM, respectively. Particularly, 5a had lower toxicity to L02 than chlorambucil (CLB) and doxorubicin (Dox), and the high selectivity was also reflected in the normal human B lymphoblast cell line (IM9). Upon investigating the mechanism of action, we found that 5a downregulated Bcl-2 and caused DNA double-stranded breaks (DSBs) to induce several genotoxic stress responses. The results of the flow cytometry assay showed that 5a was a non-specific molecule in the cell cycle. Furthermore, 5a did not affect total ROS levels but significantly improved the activity of glutathione peroxidase (GPx). Preliminary studies showed that nitrogen mustard exerted an efficient effect, and 5a can combine the advantages of artemisinin and nitrogen mustard and exhibit effects superior to either. This study showed that 5a should be further investigated as a therapeutic compound against leukemia.

Characterization of the complete chloroplast genome of Pellionia scabra (Urticaceae).

Pellionia scabra Benth. 1861 (Urticaceae) is distributed in east and southeast Asian countries, including China, Vietnam, and Japan, and has important applications in construction, medicine, and the food industry. We sequenced the genome using Illumina DNA sequencing technology. The genome was 153,220 bp long. Annotation of the genome showed that it encoded 130 genes, including 85 protein-coding genes, 37 tRNA genes, and eight rRNA genes. In addition, 15 of the genes contained a single intron and two contained two introns. Furthermore, rps12 consists of 3 exons that are expected to be trans-spliced together. Subsequent phylogenetic analysis revealed that P. scabra is closely related to the Elatostema species (i.e. E. stewardii [MZ292972], E. dissectum [MK227819], and E. laevissimum var. laevissimum [MN189961]).

KDM2A plays a dual role in regulating the expression of malignancy-related genes in esophageal squamous cell carcinoma.

KDM2A is a histone demethylase, which primarily catalyzes the demethylation of H3K36me2. Abnormal expression of KDM2A is observed in many types of cancers; however, the molecular events connected to KDM2A expression remain unclear. We report that KDM2A performs an oncogenic function in esophageal squamous cell carcinoma (ESCC) and is robustly expressed in ESCC cells. ShRNA-mediated knockdown of KDM2A resulted in a significant inhibition of the malignant phenotype of ESCC cell lines, whereas ectopic expression of KDM2A showed the opposite effect. We also analyzed the function of KDM2A using a CRISPR-CAS9 depletion system and subsequent rescue experiment, which also indicated a cancerous role of KDM2A. Interestingly, analysis of the gene expression network controlled by KDM2A using RNA-seq revealed an unexpected feature: KDM2A could induce expression of a set of well-documented oncogenic genes, including IL6 and LAT2, while simultaneously suppressing another set of oncogenes, including MAT2A and HMGCS1. Targeted inhibition of the upregulated oncogene in the KDM2A-depleted cells led to a synergistic suppressive effect on the malignant phenotype of ESCC cells. Our results revealed the dual role of KDM2A in ESCC cells, which may have therapeutic implications.

The complete chloroplast genome of Nageia fleuryi (Hickel) de Laub. (Podocarpaceae).

Nageia fleuryi (Hickel) de Laub. 1987 belongs to the genus Nageia in the family Podocarpaceae and is distributed throughout southeast Asia, including China, Vietnam, and Cambodia. It is a plant with high economic beneficial for food and construction industries. Here, we report on the complete chloroplast (cp) genome of N. fleuryi for the first time. The complete cp genome is similar to many gymnosperm plants, however, it lacks inverted repeat regions and does not possess a typical quadripartite structure. The complete cp genome is 133,870 bp in size and the overall guanine-cytosine (GC) content was found to be 37.27%. The total number of genes is 119, including 82 protein-coding genes, 33 tRNA genes, and 4 rRNA genes. Of these, 14 genes contain one intron, two genes contain two introns, and rps12 possessed a trans-splicing mechanism. Finally, the phylogenic tree demonstrated that N. fleuryi is closely related to Nageia nagi (AB830885.1 and LC572156.1).

The complete chloroplast genome of Rubus rosifolius (Rosaceae), an ornamental and medicinal plant.

Rubus rosifolius belongs to the genus Rubus in the family Rosaceae and is widely distributed globally. It has white flowers and red fruits. Moreover, it has medicinal value for diseases of the stomach and other areas. However, the complete chloroplast (cp) genome of R. rosifolius remains unclear. In the present study, we sequenced the complete cp genome of R. rosifolius (GenBank accession no. OL435124), which had a typical quadripartite structure and a size of 155,650 bp. Fifteen genes (trnK-UUU, rps16, trnG-UCC, atpF, rpoC1, trnL-UAA, trnV-UAC, petB, petD, rpl16, rpl2, ndhB, trnI-GAU, trnA-UGC, and ndhA) contained an intron, and two genes (clpP and ycf3) contained two introns. The gene rps12 showed trans-splicing. Phylogenetic analysis showed that R. rosifolius was closely related to Rubus taiwanicola, Rubus rubroangustifolius, and Rubus glandulosopunctatus.

Comprehensive Genome-Wide Identification and Expression Profiling of Eceriferum (CER) Gene Family in Passion Fruit (Passiflora edulis) Under Fusarium kyushuense and Drought Stress Conditions.

Plant surfaces are covered with cuticle wax and are the first barrier between a plant and environmental stresses. Eceriferum (CER) is an important gene family involved in wax biosynthesis and stress resistance. In this study, for the first time, 34 CER genes were identified in the passion fruit (Passiflora edulis) genome, and PeCER proteins varied in physicochemical properties. A phylogenetic tree was constructed and divided into seven clades to identify the evolutionary relationship with other plant species. Gene structure analyses revealed that conserved motifs ranged from 1 to 24, and that exons ranged from 1 to 29. The cis-element analysis provides insight into possible roles of PeCER genes in plant growth, development and stress responses. The syntenic analysis revealed that segmental (six gene pairs) and tandem (six gene pairs) gene duplication played an important role in the expansion of PeCER genes and underwent a strong purifying selection. In addition, 12 putative ped-miRNAs were identified to be targeting 16 PeCER genes, and PeCER6 was the most targeted by four miRNAs including ped-miR157a-5p, ped-miR164b-5p, ped-miR319b, and ped-miR319l. Potential transcription factors (TFs) such as ERF, AP2, MYB, and bZIP were predicted and visualized in a TF regulatory network interacting with PeCER genes. GO and KEGG annotation analysis revealed that PeCER genes were highly related to fatty acid, cutin, and wax biosynthesis, plant-pathogen interactions, and stress response pathways. The hypothesis that most PeCER proteins were predicted to localize to the plasma membrane was validated by transient expression assays of PeCER32 protein in onion epidermal cells. qRT-PCR expression results showed that most of the PeCER genes including PeCER1, PeCER11, PeCER15, PeCER17, and PeCER32 were upregulated under drought and Fusarium kyushuense stress conditions compared to controls. These findings provide a foundation for further studies on functions of PeCER genes to further facilitate the genetic modification of passion fruit wax biosynthesis and stress resistance.