MicroRNA (miRNA) are a class of endogenous non-coding small RNAs that can be involved in post-transcriptional regulation of genes. miR-10b has recently been shown to be a key regulator during ovarian atresia in pig (Sus scrofa), while phosphatase and tonic homolog genes (PTEN) play an important role in apoptosis. In order to investigate the regulatory effects of miR-10b and PTEN on apoptosis in pig ovarian granulosa cells (pGCs), the miR-10b mimic and inhibitor were transfected into pGCs to performed miR-10b overexpression and repression, respectively. The results showed that the apoptosis rate of pGCs was significantly increased after overexpression of miR-10b (P<0.01). In contrast, inhibition of miR-10b expression decreased the apoptosis of pGCs (P<0.01). To further determine the function of miR-10b, bioinformatics analysis was carried out to predict target gene of miR-10b, the result showed that 3'UTR of PTEN contained the target sites for miR-10b. By luciferase reporter assay, the firefly luciferase activity was reduced when co-transfection of pmirGLO-PTEN-WT and miR-10b mimic compared to co-transfection of pmirGLO-PTEN-WT and mimic NC (P<0.01), but no significant difference in the firefly luciferase activity between co-transfection of pmirGLO-PTEN-MUT and miR-10b mimic compared to co-transfection of pmirGLO-PTEN-MUT and mimic NC (P>0.05). In addition, co-transfection of pmirGLO and mimic NC was no significantly different compared to co-transfection of pmirGLO and miR-10b mimic (P>0.05). To better understand the mechanism of miR-10b during apoptosis of pGCs, effect of miR-10b on the expression of PTEN was verified and found that expression of PTEN gene and its protein decreased significantly after overexpression of miR-10b (P<0.01). In contrast, when miR-10b expression was inhibited by miR-10b inhibitor, the PTEN was increased compared to the inhibitor NC group. After inhibiting the expression of miR-10b, and siRNA was transfected to down-regulate the expression of PTEN gene, which partially alleviated the promoting effect of miR-10b on pGCs apoptosis. Together, it was demonstrated that miR-10b could directly bind PTEN 3'UTR and inhibit the expression level of PTEN, and miR-10b could regulate apoptosis of pGCs by inhibiting the expression of the target gene PTEN. This study demonstrates that miR-10b can regulate apoptosis by inhibiting the expression of the target gene PTEN, which provides data to support an in-depth understanding of the apoptotic mechanism of pGCs and the study of follicle development.

Thaumatin-like protein (TLP) belongs to pathogenesis-related proteins (PRPs), which plays an important role in plant resistance to biotic stresses and abiotic stress. Earlier studies have reported that TaTLP1 involved in wheat (Triticum aestivum) defense in response to Puccinia triticina (Pt). To further analyze the resistance mechanism of TaTLP1, in this study, GST-pull down and yeast two hybrid (Y2H) assays were conducted to identify the interacting proteins of TaTLP1 in wheat. 14-3-3-like protein (TaGF14) as a candidate interactor of TaTLP1 was identified by both techniques. The interaction between TaTLP1 and TaGF14 was confirmed by Y2H and co-localization. qPCR analysis showed that TaTLP1 and TaGF14 exhibited the similar expression profiles and were highly induced in the early stage of leaf rust infection. Moreover, the expression levels of both genes in the incompatible interaction were higher than that in the compatible interaction. The subcellular localization study showed that TaGF14 was localized in the nucleus. Further characteristic analysis of TaGF14 showed that it might have potential disease resistance function. This study provides a basis for further study on the molecular mechanism of TaTLP1 and participating in the defense response of wheat against leaf rust and exploring more disease resistance genes in the host and provides more excellent disease resistance resources for developing cultivated wheat resistant varieties.

The number of main stem flowers affects the yield of self-pruning tomato (Solanum lycopersicum). In order to locate the genes associated with inflorescence pruning node, a fine mapping was carried out for laying a foundation for gene cloning and functional analysis and providing evidences for the analysis of regulation mechanism. In this study, one F2 population was constructed from the cross between AXF (low nodal self-pruning tomato strain) and GXF (nodal self-pruning tomato strain) , and linkage analysis and gene localization were performed by bulked segregant analysis sequencing (BSA-seq) and molecular marker. The results showed that the associated region from 47.56~47.59 Mb was located on chromosome 2 with the size of 25 497 bp, containing 3 candidate genes (thiosulfate sulfurtransferase 18 (TST18) (GenBank No. XM_004232452.4), MLO-like protein 4 (MLO4) (GenBank No. XM_019212390.2) and 26S proteasome non- ATPase regulatory subunit 4 (PSMD4) (GenBank No. XM_010318126.3)). 118 InDel (insertion-deletion)/ CAPS (cleaved amplified polymorphism sequences)/dCAPS (derived CAPS) primers were designed for fine mapping and a dCAPS14 molecular maker closely linked with candidate gene PSMD4 was finally delimited by further analysis. qRT-PCR analysis showed that the relative expression of the candidate gene was significantly different between AXF and GXF. The PSMD4 was involved in ubiquitin-proteasome system mediated protein degradation and served as key candidate gene. PSMD4 might play an important role in the controlling of inflorescence pruning node. These results had a significance in marker-assisted self-pruning tomato breeding for ideal architecture with easy cultivation.

Pepper (Capsicum annuum) is an annual or limited perennial herb of the Solanaceae and Capsicum genus. The damaged DNA binding protein 1 (DDB1) gene has been reported in tomato (Solanum lycopersicum), Arabidopsis thaliana and other model plants, but its function in pepper is still unclear. In this study, tomato DDB1 homologous gene was cloned from pepper and named CaDDB1 (GenBank No. XM_016701551). CaDDB1 gene silencing plants were obtained by using virus-induced gene silencing (VIGS) technique, and the biological functions of CaDDB1 were preliminarily identified through observing their phenotypes. The results showed that pepper plants were extremely significantly dwarfed after CaDDB1 gene silencing (P<0.01). Simultaneously, anthocyanin content in the 3rd to 6th true leaves were extremely significantly increased (P<0.01), indicated that CaDDB1 gene affected the growth and development of pepper and played a negative role in the regulation of anthocyanin synthesis. The results of this study provide some theoretical guidance for the growth and development regulation and variety improvement of pepper.

Green prickly ash (Zanthoxylum armatum) dominated by parthenogenesis is shown to have a high proportion of male floral differentiation, which severely reduces the yield. But the underlying reason associated with stamen formation is not yet known. In previous studies, the floral organ differentiation is controlled by the ABC model in plants, and the C-class gene, named AGAMOUS (AG), plays a dominant role during the stamen and carpel differentiation processes. In this study, the coding sequence of ZaAG was isolated and identified based on our transcriptomic data and published genomic data profiles in Z. armatum, and the full CDS of ZaAG was cloned by PCR amplification technology. The results showed that the CDS length of ZaAG was 732 bp, encoding a total of 243 amino acids, including MADS domain composed of 77 amino acids and K-box domain composed of 86 amino acids. Phylogenetic analysis suggested that the AG protein was relatively conserved among the members of same family, and the ZaAG was closely related to the Rutaceae, such as Citrus clementina and C. sinensis. Additionally, a total of 148 transcription factor binding sites were predicted in ZaAG coding sequence, which were mainly involved in various reproductive growth and plant resistance pathways. The cis-elements analysis showed that the promoter region of ZaAG was enriched in the activation sites related to light signal, plant hormones, and stress. Furthermore, the ZaAG was not detected in the roots, stems, leaves, prickles and other vegetative organs, but the significantly high expression abundance was obtained during male and female flower differentiation process, especially the highest expression at the mature stage of male flowers (P<0.05), which indicated that ZaAG might be involved in the floral sex differentiation in Z. armatum. This study are of great significance for further analysis of the regulatory mechanisms related to male and female flower differentiation and breeding high-yielding germplasm in Z. armatum.

The new variety Lycoris albiflora 'Astro Girl' has gradually changing flower colors, which is a special flower color of Lycoris. Previous studies have found that the dihydroflavonol 4-reductase (DFR) genes are the key enzyme genes for the anthocyanins biosynthesis in the petals of Lycoris. In order to further study the DFR genes function, LaDFR1 and its promoter sequence were cloned by RT-PCR and Genome walking technology from petals in Lycoris chinensis×radiata 'Astro Girl' in this study. Bioinformatics and expression pattern analysis of LaDFR1 were carried out. The function of the LaDFR1 promoter was further investigated by overexpressing it in tobacco (Nicotiana tabacum) leaves. The results showed that, (1) LaDFR1 cDNA with 1 161 bp ORF was cloned, encoding 386 amino acids. The conserved domain of amino acid sequence of LaDFR1 had NADPH and substrate binding sites, which belonged to NADB Rossmann super gene family. Sequence alignment and phylogenetic analysis showed that the consistency of DFR amino acid sequence between L. chinensis×radiata 'Astro Girl' and Nelumbo nucifera was 80.43%. (2) LaDFR1 was mainly expressed in petals, followed by filaments, and the lowest expression in scape. The expression of LaDFR1 gene was the lowest in the first flowering day, and the expression increased gradually with the extension of flowering time. (3) LaDFR1 promoter sequence with 1 570 bp was obtained, which contained multiple light, abscisic acid (ABA) induction, MYB and NAC binding cis acting elements. The LaDFR1pro: PBI121 integration vector was constructed to transiently transform tobacco leaves. The GUS activity induced by light was higher than dark, which indicated that the LaDFR1 promoter had photoinduced activity. In summary, the LaDFR1 gene and its promoter sequence were successfully obtained in this study, which may be involved in the synthesis and accumulation of anthocyanins in the petals of L. chinensis×radiata 'Astro Girl'. This study can provide target genes for further study on the regulation of color changes in Lycoris albiflora and color improvement using genetic engineering.

Many varieties of peach (Prunus persica) are self-fertile, where normal pollen development is essential for yield, and abnormal pollen development affects self-pollination fertilisation and fruit set. Hence, the study of peach pollen fertility is of great industrial and theoretical importance. The sterile variety 'JiuShuo' (JS) and the fertile variety 'JiuCui' (JC) and their hybrid progeny as test material were used to study pollen fertility genes. Cytological observations indicate that pollen sterility occurred at the mononuclear microspore stage, where the cytoplasm of the tapetum could not be concentrated and degenerated. RNA-seq analysis showed that the pathways associated with the formation of programmed cell death and the outer wall of the pollen layer - sugar metabolism, phenylalanine metabolism, fatty acid biosynthesis and oxidation-reduction were significantly enriched during the tetrad and mononuclear microspore periods. The expression patterns of cytochrome P450 703A2 (PpCYP703A2) (GenBank No. XM_007220109), 4-coumarate-CoA ligase-like 1 (Pp4CL) (GenBank No. XM_007225695), and ABC transporter G family member 26 (PpABCG26) (GenBank No. XM_020565361) were examined in the fertile variety JC, the sterile variety JS, and the JS×JC progeny , using qRT-PCR to detect the expression of related genes, suggest that these three genes were involved in the regulation of pollen fertility. A new genetic resource is provided for further study of peach pollen fertility and a new theoretical basis is developed for further understanding peach pollen sterility.

Leucine rich repeat neuronal protein-1 gene (LRRN1) encodes a type Ⅰ transmembrane protein that plays an important role in neural development and regeneration. The proliferation and differentiation of bovine myoblasts directly affect the growth and development of muscle and thus affect the yield of beef. In the present study, to investigate the expression pattern of LRRN1 gene in different tissues of bovine myoblasts and its effect on the proliferation and differentiation of bovine myoblasts, to determine its major role in myogenic differentiation. qRT-PCR was used to detect the expression pattern of LRRN1 in different tissues of newborn (3 d old) and adult (24 months old) Qinchuan cattle (Bos taurus), and to detect the expression characteristics of bovine skeletal muscle myoblasts at different differentiation stages in vitro. Adenovirus mediated shRNA and pcDNA3.1(+ ) were used to construct LRRN1 gene interference and overexpression vectors, which were transfected into Qinchuan bovine myoblasts. Then EdU method was used to detect cell proliferation, and qRT- PCR and Western blot were used to detect the expression changes of proliferation marker molecules. Myogenic differentiation inducing medium was used, then the phenotype of myotube formation was observed and the expression changes of differentiation marker molecules was detected. The results showed that LRRN1 was widely expressed in various tissues of Qinchuan cattle, and the expressions of LRRN1 in kidney and longissimus dorsi muscle of newborn cattle (3 d old) and adult cattle (24 months old) were significantly higher than that in other tissues. The expression trend of LRRN1 gene was consistent with the in vitro differentiation rate of myoblasts, which increased first and then decreased, peaked at the 4th day of differentiation, and gradually decreased thereafter. The results of interfering with LRRN1 gene expression in myoblasts showed that the proportion of proliferating cells with EdU positive staining was significantly reduced (P<0.05). Proliferation marker genes CCNA2 (P<0.01), CCNB1 (P<0.05), PCNA (P<0.01), CCND2 (P<0.05) and CDK1 (P<0.05) were down-regulated, and the protein expressions of CCND2, CDK1, CCNB1 and PCNA were also decreased. Overexpression of LRRN1 significantly inhibited the proliferation of myoblasts (P<0.05). The expressions of CCNA2 (P<0.05), CCNB1 (P<0.01), PCNA (P<0.01) and CCNE2 (P<0.05) were inhibited, and the protein expressions of CCNB1, PCNA and CDK1 were also decreased. By observing the cell differentiation phenotype, it was found that interfered LRRN1 inhibited myotube formation and the expression of differentiation marker genes MYOG (P<0.01), MYF5 (P<0.01), MYF6 (P<0.01), MYH3 (P<0.01) and CKM (P<0.05), and the protein expressions of MYOG, MYF5 and MYH3 were also decreased. Overexpression of LRRN1 promoted the formation of myotube, and the gene expressions of MYH3 (P<0.01), MYOD (P<0.05), MYF5 (P<0.05) and MYF6 (P<0.05) were significantly increased, as were the protein expressions of MYOD and MYH3. Above results suggest that LRRN1 might be a positive regulator of myoblast differentiation, but its effect on myoblast proliferation needs more comprehensive and in-depth studies. Therefore, LRRN1 has a potential regulatory effect on the growth and development of muscle tissue of Qinchuan cattle, and further studies on its function can be used for molecular breeding practice of Qinchuan cattles.

Mevalonate kinase (MVK) and phosphovalonate kinase (PMVK), the key enzymes of mevalonate pathway, are considered to be closely related to inflammatory response. However, the role and mechanism of MVK and PMVK in cows (Bos taurus) with clinical mastitis (CM) is unknown. The aim of this study was to explore the expression and distribution of MVK and PMVK, and further investigate their potential function in cows with CM. The mammary gland tissues of Holstein cows in Control (C, n=3) and CM group (n=3) were collected. The expression and distribution of MVK and PMVK in cow mammary gland tissues were analyzed by hematoxylin-eosin (HE) staining, immunohistochemical (IHC) staining, immunofluorescence (IF) staining and qRT-PCR; Meanwhile, based on the data-independent acquisition (DIA) proteomic data, the role of MVK and PMVK in CM was analyzed and discussed through bioinformatics. The results showed that MVK and PMVK were mainly localized in the cytoplasm of epithelial cells; Compared with the C group, the MVK and PMVK gene and its protein levels of in CM group were extremely significantly down regulated (P<0.01); The bioinformatics analysis showed that MVK and PMVK were involved in inflammatory regulation by affecting coenzyme metabolism, amino acid and fatty acid synthesis. In conclusion, the results indicated that MVK and PMVK were correlated with the occurrence and development of CM in cows, which provides important insights into the function and regulatory mechanism of MVK and PMVK in CM of cows.

circular RNA (circRNA) plays an important role in regulating the proliferation and differentiation of preadipocytes. Previous high-throughput sequencing results showed that the expression of a circular RNA circRNA-cyclase associated actin cytoskeleton regulatory protein 2 (circCAP2) was different before and after the differentiation of Bos taurus preadipocytes. In this study, Sanger sequencing and qPCR were used to identify circCAP2 and analysed its expression pattern to research the mechanism of circCAP2 in the differentiation of preadipocytes. Furthermore, circCAP2 was transfected into preadipocytes by overexpression and interference techniques. The lipid accumulation of adipocytes was detected by oil red O staining, and the expression level of adipogenic marker genes was detected by qPCR. The results showed that circCAP2 was real presence and stably expressed, and was extremely highly expressed in mature adipocytes (P<0.01). The gain-of-function test showed that overexpression of circCAP2 significantly promoted lipid accumulation in preadipocytes, and significantly upregulated peroxisome proliferator activated receptor gamma (PPARγ), fatty acid binding protein 4 (FABP4), CCAAT/enhancer-binding protein α (C/EBPα) and C/EBPβ (P<0.05). However, the loss-of-function test showed that interference with circCAP2 significantly inhibited lipid accumulation in adipocytes, and the expression levels of adipogenic marker genes, PPARγ, FABP4, C/EBPα and C/EBPβ, were significantly down-regulated (P<0.05). In conclusion, these results suggest that circCAP2 may be a positive regulator of the differentiation of cattle preadipocytes, which can be used as a novel marker to improve fat deposition in cattle breeding. This study provides a reference for further exploring the regulatory mechanism of circCAP2 on fat deposition in cattle.

Youzhou dark goat (Capra hircus) is the genetic resources of local characteristic goat. It is of great significance to study the forming mechanism of its characteristic traits. This study collected the transcriptome data of a total of 15 samples from the heart, liver, longissimus dorsi, anterior lip and skin of Youzhou dark goat (3 samples for each tissue) for analysis. The weighted gene co-expression network analysis (WGCNA) software package was used to construct the co-expression network, and the GO and KEGG enrichment analysis was performed on the genes in the specific modules. Results showed that one specific expression module was identified in the heart and liver respectively. The genes in the heart specific blue module were mainly enriched in the citric acid cycle signal pathway and various metabolic processes. The genes in the liver specific yellow module were mainly enriched in the functions of peroxidase, amino acid metabolic process and chemical carcinogenesis. Genes in the turquoise module are mainly enriched in protein binding and other functions, genes in the green module are mainly enriched in phosphatidylinositol binding and other functions, and genes in the brown module are mainly enriched in biological processes such as cell protein metabolism. Through the CytoHubba plug-in algorithm, the 10 genes with the highest connectivity in the turquoise, blue and brown modules were defined as core genes. The tissue expression level of 6 core genes in the 3 modules were verified by qRT-PCR, and the results showed that ADCY9 and ADCY6 genes had the highest expression in heart tissue, AKT2 and MAPK3 genes were highly expressed in the anterior lip tissue, and the CAMK2G and CAMK2B genes were highly expressed in the longissimus dorsi, which were consistent with the transcriptome sequencing results. The above results have reference value for further bioinformatics research on functional genes of Youzhou dark goat. This study provides basic data for the mining, development and utilization of characteristic traits of local goat genetic resources in China.

Interleukin-17 (IL-17) is an early initiator of inflammation induced by T cells. It can promote the release of proinflammatory cytokines and stimulate the inflammatory response. At the same time, IL-17 also has the role of resisting some pathogens, so it has the role of biological adjuvant. It is unclear whether chicken (Gallus gallus) IL-17 has a biological adjuvant effect. In order to study the biological adjuvant effect of chicken IL-17, it is necessary to prepare the recombinant protein of chicken IL-17. In this study, the IL-17 cDNA sequence of chicken was amplified by reverse transcription polymerase chain reaction (RT-PCR), and its gene sequence and amino acid sequence were analyzed with biological software. In order to improve the expression level of chicken IL-17 gene in eukaryotic cells, the codon of chicken IL-17 gene was optimized according to the preferred codons of human (Homo sapiens) and chicken, and the wild-type and codon- optimized eukaryotic expression vectors were constructed respectively, and were expressed in human embryonic kidney (HEK) 293T cells. The results showed that the amplified chicken IL-17 gene sequence had 2 base differences and 1 amino acid mutation in the signal peptide sequence compared with that in GenBank. Therefore, the amino acid sequence of mature chicken IL-17 protein was not changed, indicating that the chicken IL-17 gene was polymorphic. Bioinformatics analysis showed that chicken IL-17 gene contained a typical signal peptide sequence to mediate the secretory expression of chicken IL-17, and the protein contained a N-glycosylation site. The transient expression in HEK 293T cells showed that codon optimization could significantly improve the expression level of chicken IL-17. The expression level was increased from 13 µg/ T75 Cell culture flask of wild-type IL-17 gene to 45 µg/T75 Cell culture flask of codon optimized IL-17 gene, which was 3.5 times higher, Moreover, the optimized recombinant protein could stimulate the proliferation of lymphocytes and induce IL-6 production in chicken fibroblasts. The results showed that codon optimization could obviously increase the expression of IL-17 gene in eukaryotic cells and the optimized IL-17 protein still had biological activity. This study provides favorable conditions for further study on the biological adjuvant effect of chicken IL-17.

The phenomenon of worker bee ovary development and egg laying in bumblebees indicates the decline of bee colony development. The worker bees' division of labor has shifted from active collection before egg laying to full-time egg laying, which will have a negative impact on the collection ability and pollination effect of bee colonies. In order to explore the regulation mechanism of egg-laying in workers, label- free quantitative proteomics was used to identify and compare differentially expressed proteins (DEPs) in the hemolymph of egg-laying and non-egg-laying workers of Bombus lantschouensis, then the DEPs were analyzed for GO classification and KEGG pathway enrichment, and the transcription levels of 6 randomly selected DEPs were verified by qRT-PCR. The results showed that a total of 370 DEPs were identified, including 212 up-regulated and 158 down-regulated proteins in hemolymph of egg-laying workers. GO function annotation showed that the largest number of DEGs were metabolism process (61 DEGs), followed by catalytic activity (60 DEGs) and binding (50 DEGs). DEPs enriched in detoxification, immune and antioxidant activity were up- regulated. The KEGG pathway enrichment analysis result showed that DEGs were significantly enriched in energy-related metabolism pathways including glycolysis/gluconeogenesis, carbon metabolism, fatty acid oxidation, glyoxylic acid and dicarboxylic acid metabolism, and protein synthesis-related pathways including extracellular matrix receptor interaction, ribosome, proteasome. In addition, DEPs were also significantly enriched in Notch and Hippo signaling pathway related to insect growth and development. The change trend of 6 randomly selected DEPs at transcription level was consistent with the results of proteomics. This study provides basic data for in-depth exploration of the molecular mechanism of oviposition regulation in B. lantschouensis workers, and some theoretical guidance for achieving inhibiting oviposition of workers by artificial regulation.

Compared with chemical surfactants, surfactin has the characteristics of low toxicity, environmental friendliness and reduced surface tension, which has great application prospects in agriculture, food, medicine, cosmetics and other fields, but there are problems of low yield and high cost in production. Multiple mutagenesis methods (physical, chemical and aerospace mutagenesis) were used to select and mutate Bacillus subtilis, and a genetically stable compound mutagenic strain FHYB201030 was obtained by combining blood plate, cetylpyridinium chloride-bromothymol blue (CPC-BTB) colorimetric method and high performance liquid chromatography screening through mutagenic selection in this study. Whole genome sequencing was performed on the starting strain (YUAN. 0) and the mutant strain (FUYB201030), respectively. The fermentation broth of the mutant strain (FUYB201030) was collected and purified, and then surfactin obtained was assayed for yield and in vitro antibacterial activity. The results showed that the surfactin production of FHYB201030 was 16.8 times higher than that of YUAN. 0, and the production was up to 353 mg/L. The whole genome of YUAN.0 was composed of a 5 209 013 bp complete circular chromosome and a 299 348 bp complete circular plasmid. Through preliminary analysis of the resequencing data of FHYB201030, it was found that a total of 450 SNP mutation sites in CDS of the strain were caused by complex mutagenesis, which mainly involved 28 gene sequences. Combined mutagenesis not only increased the yield of Bacillus subtilis, but also affected the growth cycle of Bacillus subtilis and improved the antibacterial ability of surfactin. Compared with YUAN. 0, the same concentration of the surfactin (40 mg/L) synthesized by FHYB201030 showed enhanced inhibitory effect on bacteria and fungi, and had significant inhibitory effect on the growth of indicator bacteria. In conclusion, the combined mutagenesis method could effectively improve the ability of Bacillus subtilis to produce surfactin and improve the activity of surfactin, which provides a basis for further research and application of surfactin.

The resources of saline-alkaline tolerant microorganisms can promote soil nutrient circulation, enhance the water reservation and salinity tolerance ability of the plant, they have important application in remediation of saline-alkaline soil. Previously, a saline-alkaline resistant bacterial strain BFL-3 was isolated, the purpose of this study was to clarify the genetic evolution relationship, saline-alkaline tolerance and mechanism of the strain. The strain BFL-3 was identified by morphology and 16S rRNA sequence analysis. The growth characteristics of strain BFL-3 under different NaCl content and pH stresses were determined by optical density measurement. Besides, the whole genome of strain BFL-3 was sequenced, followed by gene function annotation and bioinformatics analysis. qPCR was applied to verify the expression of genes related to saline-alkaline tolerance of BFL-3 in medium containing 5% NaCl and pH 9. The results showed that the strain BFL-3 was identified as Arthrobacter rhombi, it could tolerant 0.5%~10% of NaCl, the tolerance range of pH value was 5~10. The strain contained 35 scaffolds, the total sequence length was 3 476 199 bp, G+C mol% was 69.15%, coding 3 170 genes, including 50 tRNA genes and 7 rRNA genes. Gene annotation showed that strain BFL-3 was enriched in a large number of genes in amino acid and carbohydrate metabolism pathways, and a certain number of genes in ion transporter related pathways, suggesting that the strain adopts 2 strategies of solute accumulation and ion balance in response to saline-alkaline stress. When the NaCl content was 5% and the pH value was 9, the expression level of trehalose synthase gene (treS), diaminobutyrate-2-oxoglutarate transaminase gene (ectB), glutamate dehydrogenase gene (gdhA), glycine betaine/L-proline ABC transporter permease gene (proW), Trk system potassium transporter gene (trkA) and choline ABC transporter permease gene (opuBD) were up-regulated, 11.0, 6.2, 6.0, 13.1, 7.4 and 6.3 times of the control group, respectively (P<0.01). These results indicated that strain BFL-3 could alleviate the saline-alkaline stress through the synthesis and accumulation of cytocompatible substances (such as trehalose, ectoine and glutamate) and regulating ion balance in the cell. This study clarified the saline-alkaline tolerance characteristics of BFL-3, explored its saline-alkaline tolerance mechanism, provided scientific basis for the further application of this strain in remediation of saline-alkaline soil.

Homeodomain transcription factors are widely present in plant pathogenic fungi and play an important role in the morphogenesis and infestation of pathogens. In this study, bioinformatics was used to identify the Homeodomain family genes of Botrytis cinerea at the genome-wide level. The physicochemical properties, phylogenetic relationships, conserved structural domains and expression patterns during the developmental and infestation periods of the pathogenic conidia of Homeodomain were carried out. The relative expression of Homeodomain treated with NaCl and Congo red were analysed by qPCR. The results showed that there were 14 Homeodomain family genes in the genome of Botrytis cinerea, which were classified into 4 subfamilies by phylogenetic analysis; All the Homeodomain family genes of Botrytis cinerea contained the typical SANT domain of the Homeodomain family, and some of them were up-regulated during different developmental and infestation periods of the conidia. The expression levels of the Homeodomain family genes were significantly different after NaCl and Congo red treatment, indicating that the Homeodomain family genes played an important role in the growth, development, the process of infestation, and the response to stress of the pathogen. The present study provides a theoretical basis for the elucidation of the functions of the Homeodomain family genes and their mechanisms of action.

Single-cell RNA sequencing (scRNA-seq) is an advanced technique for characterizing the expression profiles of heterogeneous cells in the same tissue, and plays a crucial role in the study of cell development trajectory. Compared with animal cells, the application of scRNA-seq in plants is still in its infancy, and has been gradually applied in Arabidopsis, rice (Oryza sativa) and maize (Zea mays). This review summarized the common techniques of scRNA-seq and their applications in the research of tissue development mechanism, cells dynamic development trajectory, direction of cells differentiation and interaction. Additionally, this article analyzed the problems, challenges and development prospects of the application of scRNA-seq in plants, hoping to provide a theoretical basis for explore the mechanism of plant tissue development.

Gamete maturation and embryo development are key factors that determine mammalian reproduction. Gamete maturation and embryo development are regulated by a variety of epigenetic information. N6- methyladenine (m6A) specifically refers to the endogenous methylation modification on the sixth N atom of RNA adenine, which is the most abundant internal modification in mammalian mRNA and affects many aspects of RNA metabolism. m6A plays an important role in mammalian gamete maturation and embryo development. Abnormal modification of m6A impairs gametogenesis, sex hormone synthesis, fertility and early embryo development. This review focuses on the studies of m6A modification related enzymes and their regulation role in mammalian gamete maturation and early embryo development, which provides references for the further exploring the mechanism of m6A regulation of mammalian gamete maturation and embryo development.

Plant virus-mediated gene editing (VIGE) system can rapidly screen high-efficiency sgRNA (single guide RNA) for targeted editing of plant genes, providing a gene editing tool for the targeted creation of genetic mutant materials. In order to screen the VIGE receptor materials capable of efficiently knockout the target genes in Arabidopsis thaliana, this study using the AtBRI1 (brassinosteroid insensitive 1) and AtGL2 (GLABRA 2) as target genes, and explored the editing efficiency of different tissue-specific Cas9 overexpression (Cas9-OE) lines as VIGE receptors for these 2 genes based on the Cotton leaf crumple virus (CLCrV)-mediated VIGE system. The expression of Cas9 gene in different tissue-specific Cas9-OE lines was examined by qPCR, and it was found that Cas9 gene was stably genetically expressed in different tissue-specific Cas9-OE lines. The leaves of different tissue-specific Cas9-OE plants were inoculated with CLCrV- AtU6-26:: AtBRI1-sgRNA and CLCrV-AtU6-26:: AtGL2-sgRNA by Agrobacterium-mediated transient transformation. Mutation detection results showed that the 3 tissue-specific Cas9-OE lines as VIGE receptors could achieve targeted editing of AtBRI1 and AtGL2 genes, and different base insertions, substitutions and deletion mutation types appeared in the target sequence regions of these 2 genes, demonstrating the effectiveness of these 3 tissue-specific Cas9-OE lines as VIGE receptors. Further mutation detection was performed on each individual plant inoculated with CLCrV-AtU6-26:: AtBRI1-sgRNA and CLCrV-AtU6-26::AtGL2-sgRNA. The mutation analysis showed that ProCDC45:: Cas9 and ProYao:: Cas9 had relatively high editing efficiency for AtBRI1 and AtGL2 genes with editing efficiency of 50%~81.25%, which proved that ProCDC45:: Cas9 and ProYao:: Cas9 transgenic Arabidopsis can be used as ideal VIGE receptors for gene editing research in Arabidopsis. The study provides a basis for further improving the gene editing efficiency of the VIGE system in Arabidopsis and efficiently creating Arabidopsis mutant materials.