Rice (Oryza sativa) grain shape is a critical factor in determining the yield, appearance, nutritional quality, and commercial value of rice. Grain shape is primarily determined by genetic factors. It is crucial to enhance the quality and yield of rice by exploring and utilizing valuable gene resources related to rice grain shape, as well as analyzing the molecular mechanisms that regulate it. In this study, 410 3K rice core germplasms were used as research materials. After seed maturation, measurements were taken for grain length, grain width, and grain thickness. Genome-wide association analysis (GWAS) was conducted using the genotypes of the germplasm. Candidate genes and haplotypes were analyzed to screen for potential genes associated with these QTL. A total of 5 QTL related to grain shape were detected by GWAS, which were distributed on chromosomes 1, 3, 5, 7, and 8 of rice. The QTL controlling grain length were qGL3, qGL7, and qGL8. Among these, qGL3 was co-located with GS3, qGL7 was co-located with GL7, and qGL8 was a newly discovered QTL locus that controls grain length. The QTL controlling grain width was qGW5, which was co-located with GW5. The QTL controlling grain thickness was qGT1, which was co-located with d61. The candidate gene for qGL8 was Os08g0421800. The analysis involved aggregating different haplotypes of grain length QTL in germplasm. It was found that the favorable haplotypes of the candidate genes qGL3, qGL7, and qGL8 were present in 8 germplasms, including 'B073' and 'B081'. Additionally, the grains of these 8 germplasms were slender. This study provides new germplasm resources and theoretical support for enhancing the genetic improvement of rice grain shape.

WUSCHEL (WUS) is a homeodomain transcription factor, an evolutionary branch of the WOX family, which may play an important role in embryogenesis and bud regeneration in plant cells by regulating the expression of specific target genes. In this study, a total of 25 WUS genes were identified in the entire genome of wheat (Triticum aestivum) by bioinformatics methods. These genes were distributed unevenly on the first, second, third and the fifth homologous group of chromosomes, with tandem duplication observed on the third homologous group chromosomes. Conserved domain and structural analysis showed that motif 1 and motif 3 were present in all members of the family, and the WUS proteins clustered together showed similar gene structure according to the evolutionary tree. A total of 18 hormone response and abiotic stress response elements were identified in the promoter region of TaWUSs. Tissue expression profile analysis showed that the 25 detected TaWUSs had obvious tissue expression specificity in wheat, and most of them showed high expression in the floral reproductive organs. qRT-PCR results also confirmed that TaWUS-2A1, TaWUS-3B2, TaWUS-3D2, TaWUS-5A1, TaWUS-5B1 and TaWUS-5D1 were highly expressed in stamens, and TaWUS-3D1 was highly expressed in pistils. Especially, the high expression of TaWUS-1A, TaWUS-1B, TaWUS-1D, and TaWUS-2D1 in pistillody stamens might indicate their involvement in regulating the formation of stamen homologous transformation into pistil traits in the wheat homologous transformation sterility-1 (HTS-1). This study provides a theoretical basis for further research on the function of wheat WUS subfamily genes in wheat.

R2R3-MYB transcription factors, as the largest subfamily of the MYB transcription factor family, play a crucial role in plant growth and development, material metabolism, and other processes. Additionally, they have the potential to enhance plant stress tolerance. In a previous study, a StGAMyb-like1 gene (GenBank No. XM_015303144), homologous to AtMYB33 in Arabidopsis thaliana, was identified in potato (Solanum tuberosum). The StGAMyb-like1 gene, a member of GAMYB subgroup 18 of R2R3-MYB, is capable of responding to drought stress. In this study, the StGAMyb-like1 gene was cloned from the potato variety 'Desiree', and its CDS was 1 572 bp. Bioinformatics analysis revealed that the promoter region of this gene contained multiple hormone response elements, and its protein sequence was similar to that of tomato (Solanum lycopersicum). The expression analysis in different tissues showed that the StGAMyb-like1 gene had higher expression level in leaves. The promoter sequence of the StCYP98A3 gene, which encodes coumarate 3-hydroxylase in the lignin synthesis pathway, was analyzed and found to have MYB binding site (MBS). It was confirmed that StGAMyb-like1 binded to the StCYP98A3 promoter and activated its transcription using yeast one hybrid, dual-luciferase reporter assay and β-glucuronidase (GUS) detection. This study provides a reference basis for further exploring the function of StGAMyb-like1 gene.

Wound-induced proteins (WIPs) play an essential role in plants defence responses to pathogen infestation. In this study, the StWIP2 gene (GenBank No. NM_001288699.1) was cloned from potato (Solanum tuberosum) 'Qingshu 9', and bioinformatics analyses of the StWIP2 gene were carried out, while overexpression vectors and silencing vectors were constructed, and reverse genetics was used to characterise the function of the StWIP2 gene in the interaction with Phytophthora infestans. Bioinformatics revealed that the StWIP2 gene had a full CDS length of 636 bp and encoded a total of 211 amino acids. qPCR analysis revealed that the StWIP2 gene had the highest expression in 'Chunshu 5' and the lowest expression in diploid potato DM. The StWIP2 gene was responsive and differentially expressed under treatments of P. infestans, salicylic acid (SA), abscisic acid (ABA), and methyl jasmonate (MeJA). Overexpression of StWIP2 was able to increase the content of H₂O₂ and malondialdehyde (MDA), the activity of antioxidant enzymes, and the expression levels of pathogenesis related protein 1 (StPR1), StPR5 and ethylene responsive factor 1 (StERF1), to delay the onset of the disease in the plants and to increase the level of plant resistance to P. infestans. After silencing the StWIP2 gene, the content of H₂O₂ and MDA in plants decreased, and the activity of antioxidant enzymes decreased; The expression levels of plant defense gene 1.2 (PDF1.2), StERF1, peroxidase gene (StPOD), and phenylalanine ammonia lyase (StPAL) were reduced, while the expression of StPR1 and StPR5 were inhibited, resulting in a decrease in plant resistance to late blight. The above results indicate that the StWIP2 gene plays a positive regulatory role in the interaction with P. infestans, and the StWIP2 gene can be utilized as a candidate genetic resource for disease resistance breeding in potatoes.

PHO2 (PHOSPHATE2) functions as a negative regulator of phosphorus transport proteins and plays a role in the metabolic homeostasis of phosphorus in plants by exercising its ubiquitin-binding enzyme activity. In this study, Arabidopsis thaliana PHO2 was used as the basis to investigate properties and functions of BnaPHO2 protein, which has the highest homology with it in Brassica napus. With biological information data as a reference, the BnaPHO2 gene expression mode, subcellular localization, and yeast self -activation activity were analyzed, and the BnaPHO2 fusion protein was obtained using the prokaryotic expression system. The results showed that the BnaPHO2 protein consisted of 865 amino acids with a molecular weight of 95.7 kD, and the physicochemical properties of the protein were stable. The conserved structural region contained sites covalently bound to ubiquitin proteins and ubiquitin ligases, and phylogenetically it was closely related to cabbage (Brassica rapa) and kale (Brassica oleracea) of the same family. The results of qRT-PCR showed that the relative expression of BnaPHO2 gene was high in nutrient organs such as roots and leaves, and it positively responded to high phosphorus stress. Tobacco (Nicotiana tabacum) transient transformation results showed that BnaPHO2 was localized in the cell membrane and nucleus. Yeast self-activation assay showed that BnaPHO2 had no self-activating activity in its full length. BnaPHO2 was induced by 1 mmol/L IPTG at 28 ℃ for 16 h, and the purified proteins were obtained with N-terminal fusion of 6×His tag or C-terminal fusion of glutathione S-transferase (GST) tag. The elucidation of BnaPHO2 protein characterization and heterologous protein expression provides theoretical reference and basic information for subsequent studies of in vitro interactions and ubiquitination.

The IQ67 domain (IQD) genes encode a class of calmodulin-binding proteins that contain the conserved IQ67 domain. These proteins play important roles in regulating plant organ morphology and responding to adversity stress. In this study, 34 PvIQD genes were identified in the genome of common bean (Phaseolus vulgaris) through bioinformatics analysis. Their physicochemical properties, gene duplication events, phylogenetic evolution, gene structures, and expression patterns were analyzed. The study showed, PvIQD genes were unevenly distributed on 11 chromosomes. There were 2 segmentally duplicated gene pairs and no tandem duplicate gene pairs. The Ka/Ks ratios were all less than 1, indicating that PvIQD gene pairs have been under purification selection pressure during evlution. In the phylogenetic analysis, the PvIQD gene family members were divided into 4 subfamilies (Ⅰ~Ⅳ), with 28 PvIQD genes exhibiting a one-to-two clustering relationship with IQD genes from soybean (Glycine max). Gene structure analysis revealed that PvIQD genes contained 2~6 exons, and most PvIQD protein sequences contained motif1 and motif7. PvIQD genes in the same subfamily had similar gene structures and conserved motifs. Gln7, Arg11, Gly12, Leu14, and Arg16 in the IQ motif were highly conserved amino acids. Collinearity analysis showed that common bean had a closer genetic relationship between common bean and dicotyledonous plants (Arabidopsis thaliana and Glycine max), compared with monocotyledonous plants (Oryza sativa and Zea mays), and the closest relationship. Analysis of RNA-seq data showed that 6 PvIQD genes were highly expressed in multiple tissues at different developmental stages. The tissue-specific genes, PvIQD2 and PvIQD19, showed the highest transcript levels in green mature pods and nodules, respectively. The results of qRT-PCR analysis showed that 8 PvIQD genes exhibited distinct expression patterns in response to drought and salt stress. This study provides a reference for exploring the regulatory mechanisms of PvIQD genes in resistance to abiotic stress in common bean.

Δ9-stearoyl-ACP dehydrogenase (SAD) is a key enzyme involved in the synthesis and accumulation of unsaturated fatty acids in plants. In this study, to explore the role of the SAD enzyme in the fatty acid biosynthesis of Perilla frutescens, PfSAD3 (GENE_023515) and PfSAD4 (GENE_042129) genes were cloned and bioinformatic analysis and functional verification on them were conducted. Using the seeds of 'Jinzisu 1' as the experiment material, two full-length sequences of PfSAD3 and PfSAD4 were obtained, with the ORF lengths of 1 173 and 1 116 bp, respectively. MEGA 11.0 was used to construct evolutionary trees for the protein sequences of PfSAD3 and PfSAD4, as well as the SAD sequences of Arabidopsis thaliana and other species, it was found that PfSAD3 was closely related to LcSAD of Leucas cephalotes, and PfSAD4 was closely related to RcSAD of Ricinus communis. After constructing the yeast (Saccharomyces cerevisiae) expression vectors pYES2.0-PfSAD3 and pYES2.0-PfSAD4, PfSAD3 and PfSAD4 were transformed into the wild-type yeast strain INVSc1 and the defective yeast strain BY4389, respectively. The results showed that overexpression of PfSAD3 and PfSAD4 significantly increased the total fatty acid content and unsaturated fatty acid content in yeast (P<0.05), and restored the ability of the defective yeast to synthesize palmitoleic acid (C16:1) and oleic acid (C18:1). The substrate specificity of the PfSAD3 and PfSAD4 proteins was analyzed by adding exogenous fatty acids. The result showed that compared to PfSAD3, PfSAD4 had a more substantial substrate preference ratio for C18:0, and exhibited stronger enzyme activity. The functions of PfSAD3 and PfSAD4 were verified by overexpressing them in wild tobacco (Nicotiana tabacum). The result showed that the total fatty acid content and unsaturated fatty acid content in transgenic tobacco leaves were significantly increased (P<0.05), which was consistent with the results of yeast experiments. This study provides scientific bases and molecular targets for further elucidating the molecular regulatory mechanism of unsaturated fatty acid synthesis in oil crops such as P. frutescens, and for cultivating new oil crop varieties rich in oils through genetic engineering methods.

The sprout tumble (ST) of Pinellia ternata caused by environmental sress severely limits its industrial development, and the homeodomain-leucine zipper (HD-Zip) family genes are widely involved in the regulation of plant stress response. To explore the HD-Zip genes involved in the regulation of ST in P. ternata, the HD-Zip family genes of P. ternata were identified and analyzed at the genome level in this study. It was found that 17 HD-Zip family genes were identified in P. ternata, among them, 15 of which were attached to 9 chromosomes. Evolutionary analysis showed that the identified 17 HD-Zip family members included 9 HD-Zip Ⅰ subfamily members and 8 HD-Zip Ⅱ subfamily members, all of which had highly conserved HD domains (WFQNRR) in their protein sequences. Subcellular localization prediction showed that all 17 members of the HD-Zip family were located in the nucleus, and collinearity analysis showed that 8 PtHDZ genes formed 4 pairs of homologous gene pairs, and 1 genes had a collinearity relationship with rice (Oryza sativa). The elements of hormones response, stress response, and transcription factor binding sites were contained in the promoters of PtHDZ genes via the prediction analysis of cis acting elements. Organizational expression pattern analysis revealed that the PtHDZ family genes showed differential expression in different tissues, most of which were highly expressed in roots and tubers. And the expression of 14 PtHDZs were found to be induced while 2 PtHDZs were inhibited by high temperature. This study provides genetic resources for analyzing the mechanism of ST and molecular breeding in P. ternata.

The proliferation, apoptosis and steroid hormone synthesis of granulosa cells play important roles in follicle development, and these functions are regulated by signaling pathways. In this study, the transcriptome sequencing of granulosa cells from the first large follicle F1 and the second large follicle F2 in bovine (Bos taurus) first follicle wave pre-deviation (PD) and onset of diameter deviation (OD) were separately performed to obtain the expression patterns of genes in Wnt/β-catenin and Notch signaling pathways, cell cycle, apoptosis and steroid hormone synthesis, and to validate the expression of key components of the 2 pathways. The results showed that the genes of key components of the 2 pathways with high expression in PDF1, PDF2, ODF1 and ODF2 follicular granulosa cells were wingless-type MMTV integration site family, member 10a (Wnt10a), Wnt2b, frizzled3 (Fzd3), low density lipoprotein-related receptor 8 (LRP8), dishevelled 3 (Dvl3), Axin1, glycogen synthase 3β (GSK3β), catenin beta 1 (CTNNB1), transcription factor 7-like 2 (TCF7L2), lymphoid enhancer factor1 (LEF1), B-cell CLL/Lymphoma 9 (BCL9), Notch2, Dll4 and Hes related with YRPW motif2 (Hey2). Cell cycle-related genes with high expression were cyclin2 (CCND2), cyclindependent kinase4 (CDK4), cyclin-dependent kinase inhibitor 1A (CDKN1A), CDKN1B. The apoptosis of granulosa cells was mainly regulated by BCL2-associated X (Bax) in the mitochondrial apoptosis pathway and executed by Caspase3 and Caspase6. The expression of the steroid genes were 3β-hydroxysteroid dehydrogenase (3β-HSD), cytochrome P450 family 11 subfamily A member 1 (CYP11A1), and CYP19A1 were elevated, and E2 synthesis ability of OD follicular granulosa cells was increased compared with that of PD follicles. Wnt2 protein was localized in follicular granulosa cells in deviation process and its expression was significantly higher in OD follicular granulosa cells (P<0.05); Wnt2b protein was localized in follicular granulosa cells and theca cells and its expression was extremely significant lower in OD follicular granulosa cells (P<0.01). The mRNA expression of CTNNB1, LEF1, Notch2 and Hey2 were all extremely significantly increased in OD follicular granulosa cells (P<0.01). This study showed that the gene expression in F1 and F2 follicular granulosa cells during the deviation of bovine first follicular wave was specific, and the gene expression of key components in Wnt/β-catenin and Notch signaling pathway had the same trend, which provides a reference basis for the molecular regulatory mechanism of granulosa cell function during follicular development.

Micro RNA (miRNA) is involved in numerous physiological activities in mammals, with miR-133a playing a pivotal regulatory role in placental and uterine growth and development. miR-133a_R+1 and ALDH5A1 are differentially expressed in placental tissues of cows (Bos taurus) with retained placenta and may have a potential targeting regulatory relationship through sequencing and target gene prediction software. In this study, normal fetal placental tissue, RP placental tissue, and bovine endometrial epithelial cells (BEND) were used as experimental materials. Hematoxylin eosin (HE) staining was used to observe the pathological changes of placental tissue, and qRT-PCR was used to detect expression of miR-133a_R+1 and aldehyde dehydrogenase 5 family member A1 (ALDH5A1) in placental tissue. Immunohistochemistry (IHC) was used to detect the distribution of ALDH5A1 protein in placental tissue. Optimal transfection conditions was 100 nmol/L 133a_R+1 mimic for 12 h to establish a BEND cell model overexpressing miR-133a_R+1. qRT-PCR and Western blot were used to detect the expression of ALDH5A1 gene and its protein in the cells. The results showed that RP placental tissue villi were short, sparse, and fragmented. The expression level of miR-133a_R+1 mRNA in RP placenta tissue was extremely significantly up-regulated (P<0.01), ALDH5A1 was expressed in connective tissue and cytotrophoblast, and the expression level was extremely significantly down-regulated (P<0.01). The expression levels of ALDH5A1 gene and its protein in BEND cells transfected with miR-133a_R+1 mimic were extremely significantly downregulated (P<0.01). In summary, miR-133a_R+1 affected cow RP by targeting the expression of ALDH5A1 gene and its protein. This study provides corresponding theoretical basis for further exploring the mechanism of miRNA action on dairy cow RP.

Low temperature is one of the factors affecting the efficiency of pig production in the northern region of China, which leads to the decline of pig production performance and the increase of piglet disease and death rate, and seriously restricts the efficient development of China's pig industry. The aim of this study was to analyzing the effects of low temperature on liver antioxidant capacity and lipid metabolism in Hezuo pigs (Sus scrofa). 75-day-old healthy Hezuo pigs were selected and randomly divided into low-temperature group (CH) and normothermic control group (NH) after 7 d of room temperature rearing. The CH was reared in a room equipped with a temperature control device (-15±2) ℃, and the NH was reared in a room at temperature (23±2) ℃. The precess period was 15 d. Blood was collected from fasting anterior vena cava on the 0, 5, 10, and 15 d of the precess period, and liver tissues were collected from slaughter on the 15 d; serum antioxidant indexes and hepatic lipid metabolites were detected by ELISA, and lipid metabolism-related genes were detected by qPCR. The results showed that with the prolongation of low temperature treatment time, the serum total antioxidant capacity (T-AOC) and malondialdehyde (MDA) content of Hezuo pigs showed a trend of increasing and then decreasing, and the T-AOC activity reached the highest value of 1.94 on 10 d, which was significantly higher than the other treatment (P<0.05), and the MDA activity was the highest on 5 d, which was significant higher than the 10 and 15 d (P<0.05), The activities of serum superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) showed an increasing trend, in which SOD was extremely significant (P<0.01) after 10 d of low temperature treatment, GSH-Px and CAT activities were extremely significant (P<0.01) after 5 d of low temperature treatment. The contents of protein carbonyl (PCO) and 8-hydroxy-2deoxyguanosine (8-OHdG), markers of oxidative damage in pig livers, were extremely significantly elevated (P<0.01) in the CH compared with the NH. The activities of fatty acid synthase (FAS) and acetyl CoA carboxylase (ACC), the enzymes related to lipid metabolism, were significantly reduced (P<0.05) in pig livers exposed to low temperature. The activities of lipoprotein lipase (LPL), hepaticlipase (HL), alanine aminotransferase (ALT), and stearoyl-CoA desaturase 1 (SCD1) were all significantly reduced (P<0.01); carnitine palmitoyltransferase enzyme (CPT) activity was extremely significantly elevated (P<0.01). The serum triglyceride (TG) content of the low-temperature Hezuo pigs was the highest at 0.82 on 5 d, and significantly decreased to 0.61 on 15 d (P<0.05). The expression of hepatic lipid metabolism-related genes sterol regulatory element-binding protein-1c (SREBP-1c) and peroxisome proliferators-activated receptors α (PPARα) was significantly reduced (P<0.05), and p300/CBP associated factor (PCAF) expression was extremely significantly increased (P<0.01). In summary, the low temperature caused oxidative damage to the liver of Hezuo pigs, and Hezuo pigs could improve their cold adaptation by enhancing hepatic lipid metabolism when they were in a cold environment. This study provides a reference basis for further exploring the mechanism of the formation of cold-resistant traits in Hezuo pigs.

China's annual soybean production is insufficient to meet the demands of production and daily life, resulting in a heavy reliance on imports. Internationally, the price of soybeans continues to soar. However, substituting wheat for corn can lead to a reduction in soybean meal usage. ε-Poly-L-lysine hydrochloride (ε-PLH) is a natural preservative and antimicrobial agent commonly employed in the food industry and other related fields. This study aimed to investigate the effects of adding ε-PLH to wheat-soybean meal feed on the production performance, egg quality, and hepatic lipid metabolism of laying hens (Gallus gallus domesticus). The experiment involved a total of 120 Hy-line brown laying hens at 21 weeks of age, which were randomly divided into 4 groups. Each group comprised 5 replicates, with 6 chickens per replicate. The groups were designated as follows: Control group, wheat meal group, wheat meal+0.01% ε-PLH group, and wheat meal+0.05% ε-PLH group. The hens were fed with the respective diets for 3 weeks during the adaptation period, followed by a formal experimental period of 12 weeks. The results indicated that there were no significant differences in laying rate and feed to egg ratio among the control group and the three treatment groups. However, the average egg weight of the wheat meal+0.01% ε-PLH group and the wheat meal+0.05% ε-PLH group was significantly higher than that of the control group (P<0.05). The egg shape index, albumen height, and haugh unit of the wheat meal group, wheat meal+0.01% ε-PLH group, and wheat meal+0.05% ε-PLH group were significantly higher than those of the control group (P<0.05). Additionally, the eggshell thickness of the wheat meal+0.01% ε-PLH group was significantly higher than that of the control group and the wheat meal group (P<0.05). Wheat meal+0.01% ε-PLH group could significantly up-regulate the expression of lipid metabolism related genes such as peroxisome proliferator-activated receptor gamma (PPARG), fatty acid desaturase 2 (FADS2) and fatty acid binding protein 1 (FABP1)(P<0.05). Thus, it could be inferred that substituting wheat for corn and supplementing with ε-PLH can improve the production performance and egg quality of laying hens, alleviate hepatic lipid deposition. This study provides theoretical and practical support for the application of ε-PLH in poultry farming and offers alternative solutions for national policies aiming to reduce the use of corn and soybean meal.

The integral membrane protein 2A (ITM2A) plays an important role in various biological processes such as muscle development. Previous research results showed that the ITM2A gene is significantly related to chicken (Gallus gallus) body weight. However, the biological function of ITM2A in the proliferation and differentiation process of chicken skeletal muscle is not clear. In this study, bioinformatics analysis of chicken ITM2A protein was performed using online software. Six 7-day-old Arbor Acres Plus (AA) broilers were selected, and samples of heart, liver, spleen, lung, kidney, breast muscle, leg muscle were collected for tissue expression profile analysis. Breast muscle and leg muscle tissues at different developmental stages were selected for temporal expression analysis. The relative expression of ITM2A were detected using qPCR, and the ITM2A gene coding sequence was amplified using PCR technology to construct a recombinant plasmid for overexpression. Chicken primary myoblasts (CPMs) were isolated, and the relative expression of ITM2A at different stages of CPM induction differentiation were measured. The biological function of ITM2A was evaluated by assessing the expression of cell proliferation and differentiation marker genes in CPM cells overexpressing ITM2A. The bioinformatics analysis results showed that the ITM2A protein consisted of 251 amino acid residues, predominantly composed of α-helices (38.25%) and β-sheets (37.75%). It was an unstable, hydrophilic, non-secretory protein with a transmembrane structure, mainly located on the cell membrane, and might interact with proteins such as fin bud initiation factor homolog (FIBIN), ribosomal protein lateral stalk subunit P0 (RPLP0), and catenin alpha like 1 (CTNNAL1). It exhibited the highest homology with turkeys (Meleagris gallopavo) and the closest evolutionary relationship. Tissue expression analysis revealed that ITM2A was widely expressed in the examined tissues, with the highest expression in the lung and spleen and the lowest expression in pectoral and leg muscles. Temporal expression analysis showed that ITM2A expression gradually decreased with increasing days post-hatch, reaching its lowest point at 1 week post-hatch. Compared to the control group, the expression of ITM2A in the experimental group was extremely significantly increased (P<0.01). There were no significant changes in the relative expression levels of cell proliferation-related marker genes, while the expression levels of cell differentiation-related marker genes, such as myosin heavy chain (MYHC), myoblast determination (MYOD), and myomarker (MYMK), were significantly upregulated (P<0.05) in the overexpression group. The expression of the ITM2A gene showed a decreasing trend during CPM proliferation and an increasing trend with induction days. This study demonstrated that ITM2A could promote the differentiation of chicken myoblasts, providing fundamental information for further elucidating the molecular mechanisms through which ITM2A influences muscle development in chickens.

This study was aim to analyze the testis tissue from healthy adult male ducks (Anas platyrhynchos) with differences in testicular size and semen production by RNA-seq and bioinformatics. The differentially expressed genes (DEGs) and signaling pathways related to testicular size and spermatogenesis were screened to provide data for supporting male duck breeding. In this study, 40 male ducks (230-day-old) were used to collect testes tissues according to testis size (light weight of testes, GS; heavy weight of testes, GL) and semen production (less semen volume, GF; excessive semen volume, GM). Collect the testicular tissues and use the BGIseq platform for transcriptomic mRNA sequencing, GO and KEGG functional annotation and enrichment analysis were used to analyze the candidate DEGs. The expression of DEGs were verified by qRT-PCR. The results showed that there was a significant difference in testicular size among male ducks with no significant difference in body weight, there was also a significant difference in the production of semen after artificial training. A total of 383 DEGs were analyzed in different semen production and different testis size groups, and 36 DEGs were screened among different groups. The results of qRT-PCR showed that the result of transcriptome sequencing was accurate and reliable. GO and KEGG analysis showed that DEGs was significantly enriched in signal transduction, cell membrane, membrane composition, plasma membrane, G protein-coupled receptor activity, positive regulation of cell division and cAMP signal, pI3K-AKT signal, calcium signal pathway and other related pathways, genes GPR50, GPR132, GRER1, IGF1, TACR3, HTR1F, FGF19, FGF3, GABRG1, ADCY1 were enriched in these pathways, which might be closely related to testicular function. This study screened the genes related different testicular size and spermatogenic capacity in ducks, which provides a theoretical basis for the genetic breeding of testicular function.

As a rare natural heritage site, the ancient tea plantation of Qiongying mountain holds significant ecological, cultural, and economic value. Its sustainable development is inseparable from a healthy soil micro-ecological environment. To investigate the structural characteristics of soil bacterial communities in the ancient tea plantation of Qiongying mountain, Fengqing county, Yunnan province, this study focused on the rhizosphere and bulk soils of the ancient tea plantation on Qiongying Mountain. Using 16S rRNA gene amplicon sequencing technology, the diversity and structural characteristics of the soil bacterial communities and their relationship with soil chemical properties were analyzed. The results showed that: (1) Alpha diversity analysis indicated the bacterial community abundances in the rhizosphere and bulk soils of the valley were significantly higher than that in the rhizosphere and bulk soils of the mountaintop; there were no significant differences in the abundance or diversity of bacterial communities between rhizosphere and bulk soils at the mountaintop and valley. (2) The dominant phyla were Acidobacteriota (21.15%~26.76%) and Proteobacteria (15.06%~20.79%), and the dominant genera were Candidatus_Solibacter (2.6%~3.6%), Acidibacter (2.6%~3.4%) and Bryobacter (2.2%~2.6%). (3) Beta diversity analysis showed that the composition and structure of bacterial communities were similar but had certain differences. (4) Redundancy analysis (RDA) showed that soil organic matter was the main environmental factor affecting the composition and structure of bacterial community in rhizosphere soil. In conclusion, this study preliminarily explored the characteristics of the bacterial community structure in the rhizosphere and bulk soils of ancient tea trees at the mountaintop and valley of the Qiongying Mountain tea plantation. These findings can provide a reference basis for the protection and utilization of ancient tea trees in tea plantations.

Damping-off of cucumber caused by Pythium aphanidermatum is a serious seedling disease in cucumber (Cucumis sativus) production, which seriously affects the yield and quality of cucumber. In order to obtain a biocontrol strain with antagonistic effect against damping-off of cucumber, in this study, Bacillus spp. were isolated from the inter-root soil of healthy cucumber using a gradient dilution plating method, and obtained the biocontrol strain ZF513 with the best preventive effect against damping-off of cucumber, which amounted to 86.64%, through potted plants in greenhouse. After morphological, physiological and biochemical tests, unique carbon source utilization determination and molecular identification, it was determined that the biocontrol strain ZF513 was Bacillus velezensis. Plate confrontation method was used to determine that strain ZF513 had obvious antagonistic effect on 8 kinds of plant pathogens (Pythium aphanidermatum, Rhizoctonia solani, Fusarium oxysporum, F. solani, Corynespora cassiicola, Phytophthora capsici, Botrytis cinerea, Mphylium solani) with a broad-spectrum bacterial inhibition ability, in which the inhibition of P. aphanidermatum reached 65.23%. The results of pot test showed that strain ZF513 had obvious growth-promoting effect on cucumber plants, and the highest prevention effect on damping-off of cucumber was 72.61% when the concentration was 1×10⁸ cfu/mL and the dosage was 10 mL/plant, and the strain could stabilize and colonize the inter-root soil of cucumber with the concentration of 10⁵ cfu/g, which could protect the cucumber plants from disease invasion during the period of seedlings. In conclusion, strain ZF513 has potential application value in the control of damping-off of cucumber.

Citrus sour rot is the common citrus (Citrus reticulata) postharvest disease which can cause fruit rot and acidity, seriously affect the storage effect. Microbial control of citrus acid rot has the advantages of being green, environmentally friendly, and no drug resistant, and is the direction for the healthy development of agriculture. In this study, the effect of Streptomyces pactum Act12 against the sour rot pathogen Geotrichum candidum was evaluated by plate confrontation culture method and growth rate method, the control effect was verified by acupuncture inoculation method, and the effects of filtrate on citrus resistant enzymes, resistant substances, and fruit quality were determined. The results showed that the inhibition rate of Act12 against Geotrichum candidum was 76.04%, and the inhibition rate of 20% fermentation filtrate against Geotrichum candidum reached 97.17%. Act12 had antibacterial activity against 8 types of pathogens derived from fruit. The filtrate significantly decreased the disease index of citrus sour rot (P<0.05), showing control effect of 88.20%. After treatment with 20% fermentation filtrate, the activities of fruit resistance enzymes - polyphenol oxidase (PPO), phenylalanine ammonia-lyase (PAL), chitinase (CHI), and glucanase (GLU), as well as the total phenolic and flavonoid content of the fruit peel were significantly increased (P<0.05), while the content of soluble solids and reducing sugars was significantly increased (P<0.05). The titratable acidity was significantly decreased (P<0.05), and the fruit quality was improved. This study provides theoretical reference and new resources for the biological control of postharvest sour rot in citrus fruits.

Rabbit hemorrhagic disease virus type 2 (RHDV2) seriously threatens the safety of Oryctolagus cuniculus breeding and production. This study aimed to identify the pathogen of suspected RHDV2 infection in a meat rabbit farm in Zhejiang province, and clarify the pathogenic characteristics of isolated strain and its influence on the structure of rabbit intestinal flora. The liver samples of dead rabbits were collected firstly, the capsid protein VP60 gene of RHDV2 was amplified by RT-PCR, and pathogens were identified by sequencing and phylogenetic tree analysis. Animal regression test was used to clarify the infection and pathogenic characteristics of isolated strain, and 16S rDNA sequencing technology was used to analyze the structural changes of cecal microbiota between the blank control group and RHDV2 group. The results of the sequencing and homology analysis showed that the target fragment amplified by RT-PCR was RHDV2 VP60 partial sequence (GenBank No. PQ215520), which was homologous to China RHDV2 strain (GenBank No. OQ570964.1) with a consistency of 98.66%. The results of animal regression test showed that all 50-day-old rabbits died within 48 h of infection, and presented typical clinical symptoms such as mouth and nose bleeding. Staining of tissue sections showed disorders of liver tissue structure, and there were obvious flake necrosis hepatocytes in the liver parenchyma, more inflammatory cell infiltration in the alveolar wall, and large-scale pulmonary edema. The liver RNA of infected rabbits was extracted for PCR amplification and sequencing, and the results showed that it was consistent with the isolated strain from the diseased rabbit farm. The results of 16S rDNA high-throughput sequencing showed significant differences in the structure and relative abundance of intestinal flora between the RHDV2 group and healthy control group, and 13 species with significant differences were screened at the genus level, indicating that RHDV2 infection altered the original microbiome of the rabbit's intestine. This study explored the pathogenicity of RHDV2 isolates in Zhejiang province for the first time, which provides a reference for the prevention, control, diagnosis and treatment of RHDV2.

Bovine viral diarrhea (BVD) is an infectious disease of cattle worldwide caused by Bovine viral diarrhea virus (BVDV), which has brought huge economic losses to animal husbandry. The previous study found that knocked down the expression of signal peptidase complex subunit 1 (SPCS1) in bovine kidney cells (Madin-Darby Bovine Kidney, MDBK) could inhibit the replication of BVDV effectively. In order to explore its molecular mechanism, this study collected exosomes from the supernatant of SPCS1 knockdown (KD) cells infected with BVDV, and transmission electron microscopy, Western blot and Nanoparticle Tracking Analysis were used to characterize exosomes. qPCR was used to analysis BVDV 5' untranslated region (UTR) RNA levels in the exosomes. The exosomes were infected with MDBK; BVDV 5'UTR RNA levels were detected by qPCR; The cytopathic effect (CPE) was observed by inverted microscope, and the change of virus titer was determined by Reed-Muench method. The exosomes were observed as disk-like vesicles under transmission electron microscopy, Western blot detected exosomes, the volume characteristic protein TSG101 showed a peak value at about 100 nm, indicating exosomes were successfully extracted. Western blot showed that the expression of exosome TSG101 protein in SPCS1 KD cells infected with BVDV was significantly decreased compared with the control cells treated with empty vector (Scramble cells)(P<0.001). qPCR detection showed that the level of BVDV 5'UTR mRNA in exosomes was significantly decreased (P<0.001). BVDV 5'UTR RNA levels of SPCS1 KD cell derived exosomes infected with MDBK were significantly lower than those infected with Scramble cell (P<0.001), the CPE phenomenon was delayed and weakened, and the viral titer of the progeny was significantly decreased (P< 0.001). The results showed that knockdown SPCS1 affected the replication of BVDV through exosome pathway. The research results provide a new antiviral target and theoretical basis for the prevention and control of BVDV.

Native bacteria are resident in the animal body (mainly refers to the gut), which are non-pathogenic to the host animal, and even beneficial to the host animal, and play an important role in the health and physiological regulation of the host animal. In recent years, in the livestock or aquaculture industry, the exploration and application of animal native bacteria resources has gradually emerged, and there are great prospects. It has good application prospects in promoting the growth of livestock and poultry, improving feed conversion efficiency, preventing and treating intestinal pathogens, as well as in developing oral vaccines or biological agents for livestock and poultry. This article provided a comprehensive overview of the natural advantages of using native bacteria in the animal husbandry, including their direct use and the development of vaccines or biological agents specifically for livestock and aquaculture. The current research status, both domestically and internationally, on the application of native bacteria in the animal husbandry and the development of related ancillary products were summarized. Additionally, the current challenges, key areas, and future development directions for the application of native bacterial resources in animal husbandry were analyzed to promote their use, reduce dependence on antibiotics, and support the sustainable development of the animal husbandry.

Establishment of plant regeneration system is essential for plant genetic transformation, and the use of antibiotics in plant genetic transformation systems is detrimental to cell differentiation, thus exacerbating the difficulty of establishing plant regeneration systems. Overexpression of the DELLA protein GbGAI4 (GAI: gibberellic acid insensitive) gene confers plant dwarfism, which is a visible and easily recognizable morphological phenotype in plants, and is expected to be used as a morphological screening marker in plant genetic transformation systems. In this study, pTA7002-GbGAI4 vector was constructed, the embryo tips of island cotton (Gossypium barbadense) 'Xinhai16' was infiltrated by Agrobacterium-mediated method, the expression of GbGAI4 gene was induced by dexamethasone, and then preliminarily screened out the transformed plants with dwarf characters, and further identified the transgenic plants with RT-PCR technology, so as to establish a noval genetic transformation system in cotton. Based on this, the growth-regulating factors (GRF) GRF4 was selected as the example gene for transgenic transformation based on the GbGAI4-mediated genetic transformation system, and a gene transformation rate of (4±0.6)% was obtained. This study provides a new genetic transformation platform for molecular breeding of cotton.