SQUAMOSA promoter-binding protein like (SPL) gene family plays an important role in growth and development of embryos, buds, leaves, flowers, fruits and other organs in plants. The members of the SPL gene family in peach were identified and their expression characteristics in different tissues and fruit development processes were analyzed in this study. The results showed that 25 PrupeSPL gene family members were identified from the peach genome. Except that PrupeSPL16 lacked the Zn-2 structure, all other PrupeSPL genes contain an SBP conserved domain, two zinc finger structures and a nuclear localization signal. Most members of the SPL gene family were expressed in different tissues, and most members had higher expression in young fruit. With the fruit growth and development, the expression level of PrupeSPL1, 3, 4, 7, 8, 10, 13, 14, 16~21, 24, 25 significantly decreased, revealing that these members might be involved in the regulation of peach young fruit. During the fruit ripening and softening, the expression of PrupeSPL1~4, 7, 8, 10, 16, 17, 19, 21 showed a significant up-regulation trend, indicating that these members might be related to peach fruit ripening, softening and senescence. In this study, the PrupeSPL gene family was systematically analyzed in peach, and the PrupeSPL members that might be involved in regulating the growth, ripening and softening process of peach fruit were screened. This study provides basic data for further elucidating the function of SPL gene during peach fruit development.

Porcine reproductive and respiratory syndrome (PRRS) is one of the severe infectious diseases in the pig (Sus scrofa) industry. Cluster of differentiation 163 (CD163) is the receptor of Porcine reproductive and respiratory syndrome virus (PRRSV), and scavenger receptor cysteine-rich domain 5 (SRCR5) coded by exon 7 of CD163 is essential for PRRSV infection. In this study, CRISPR/Cas9 technology was used to produce anti-PRRSV cloned pigs. Two pairs of small guide RNA (sgRNA) targeting the intron 6 and intron 7 of CD163 gene were designed and assembled, then transfected into pig embryonic fibroblast cells (PEFs) with Cas9 expression vector. Thirty-two cell colonies were selected by limited dilution method. After PCR amplification and sequencing identification, the PEFs with deletion of SRCR5 sequence were obtained, and the efficiency of homozygous knock-out was 6.25%. Then both of them were selected as nuclear donors for somatic cell nuclear transfer (SCNT), and one gene-modified cloned pig successfully survived at last. Taken together, the present study precisely deleted functional domain of target gene, which could provide methodology reference for related researches, and the obtained CD163 gene-modified pig could be used for further study on disease resistance breeding.

The growth rate and carcass quality of beef cattle (Bos taurus) have important economic value in beef cattle industry. Angiopoietin-like protein 3 gene (ANGPTL3) is known to be a kind of secreted protein, and plays an important role in lipid, glucose and energy metabolisms. A previous study reported that there were g.-38T>C and g.509A>G mutations identified in ANGPTL3 gene, which were associated with growth and carcass traits in beef cattle. The aims of this study were to investigate the genetic diversity of g.-38T>C and g.509A>G mutations in different cattle populations, and confirm their association with growth and carcass traits in Qinchuan cattle (B. taurus). Total of 599 cattle sample (including 384 Qinchuan cattle, 24 Luxi cattle, 41 Mongolia cattle from Inner Mongolia, 50 Mongolia cattle from Mongolia, 50 Wuling cattle and 50 Longlin cattle) were collected for this study. All of 599 samples were genotyped by direct sequencing and MassARRAY technologies, and then the analyses of genetic diversity in 6 cattle populations and association with growth and carcass traits in Qinchuan cattle were performed. The results showed that the genotype frequency of g.-38T>C and g.509A>G in ANGPTL3 were consistent with Hardy-Weinberg equilibrium (P＜0.05) in each of 6 cattle populations. There were 3 genotypes TT (0.695), TC (0.261) and CC (0.044) of g.-38T>C mutation in Qinchuan cattle population. Association analysis results showed that g.-38T>C SNP was significantly associated with body length, hip width, chest circumference and back fat thickness (P＜0.05), as well as ultrasound loin muscle area and intramuscular fat content (P＜0.01) in Qinchuan cattle. Furthermore, there were 3 genotypes AA (0.690), AG (0.279) and GG (0.031) of g.509A>G mutation in Qinchuan cattle population. Association analysis results showed that g.509A>G SNP was significantly associated with body length, pin bone width, hip width and intramuscular fat content (P＜0.05), as well as chest circumference and ultrasound loin muscle area (P＜0.01). The combination analysis of two SNPs showed that the individuals with TC and AG genotypes had significantly greater ultrasound loin muscle area and intramuscular fat content than individuals with TT and AA (P＜0.05). Thus, these results suggested that g.-38T>C and g.509A>G in ANGPTL3 gene could be an effective molecular marker for marker-assisted breeding of Qinchuan cattle, and these results could provide scientific basis for genetic improvement of Qinchuan cattle.

SBP-box (SQUAMOSA PROMOTER BINDING PROTEIN box) gene family encode a class of plant-specific SBP transcription factors, which are widely distributed in plants and play important functional regulation in plant growth, development and reproduction, including embryogenesis, secondary metabolites biosynthesis, stress responses, etc. Therefore, it has application value in crop improvement. This study used bioinformatics and quantitative real-time PCR (qRT-PCR) to identify and analyze the expression of Sorghum bicolor SbSBP15 gene, and the recombinant SbSBP15 protein was obtained by Escherichia coli prokaryotic expression system. The results showed that the full length of SbSBP15 gene (GenBank No. MT634207) was 1 227 bp, encoding 408 amino acids. Bioinformatics analysis showed that SbSBP15 protein contained a typical SBP domain with a relative molecular weight of 42.19 kD and the isoelectric point was 8.87. Phylogenetic results showed the closest family relationship with maize (Zea mays). Subcellular localization was mainly in the nucleus, and it had strong hydrophilicity. Besides, protein interaction analysis demonstrated that SbSBP15 might interact with protein kinase. qRT-PCR analysis demonstrated that SbSBP15 was mainly expressed in the stalk, and drought and salt stresses could induce SbSBP15 expression. Furthermore, the expression level of SbSBP15 was also enhanced after plant hormone indole acetic acid (IAA) treatment, however, salicylic acid (SA) reduced the expression of SbSBP15. The optimal protein expression of SbSBP15 were in E. coli Rosetta (DE3) strain, inducing temperature at 25 ℃ and isopropyl-β-D-thiogalactopyranoside (IPTG) concentration of 0.8 mmol/L. This study provides a candidate gene for SbSBP15-based molecular genetic study of sorghum, and further provides a basis for studying the biological function of plant SBP15 gene.

The single transgenic Bacillus thuringiensis (Bt) gene Cry1Ab/Cry1Ac insect-resistant cotton (Gossypium) is a large-scale planted crop in China. With the increase of planting time and area, the resistance of pests to Bt toxin protein increases, new Bt gene research work is necessary. Cry5Aa is a novel Bt insecticidal gene with resistance to both Lepidoptera and Nematodes, was successfully transferred into cotton by Cotton Research Institute of Hunan Agricultural University and a stable genetic 'JX0010', 'JX0020' strain was obtained. In this study, the Cry5Aa full-length gene prokaryotic expression vector was constructed, the prokaryotic expression induction conditions were optimized, and its expression in the Bt-Cry5Aa gene of transgenic cotton line was identified. The results showed that the protein expression was the highest when induced at 37 ℃ with 220 r/min 1.0 mmol/L isopropyl-beta-D-thiogalactopyranoside (IPTG) for 4 h. Through Western blot experiment, it was found that the molecular weight of the protein was 79 kD, which was consistent with the theoretical value; at the same time, the protein existed in the form of inclusion body and was purified by Glutathione-Sepharose 4B small particle affinity chromatography. The identification experiment of transgenic cotton lines with Bt-Cry5Aa gene showed that Cry5Aa gene could express insecticidal protein in transgenic line 'JX0010', 'JX0020', and the expression of Cry5Aa toxin protein of 'JX0010' was higher than that of 'JX0020' in different periods and different parts of cotton. The expression in bud stage was the highest, and the spatial distribution was the highest in leaves. In this study, the prokaryotic expression vector of Bt-Cry5Aa insecticidal gene was constructed, the purified protein of Cry5Aa was obtained, and its expression in cotton was preliminarily identified, which provides a theoretical basis for the further application of Bt-Cry5Aa insecticidal gene in cotton.

Lipoxygenases (LOXs) are a class of non-heme and iron-containing fatty acid dioxygenases that play a key role in the lipid oxidation process and have an impact on the seed aging process. Onion (Allium cepa) seeds are short-lived, which have a serious impact on actual production research. In order to study the effect of LOX gene on the development and deterioration of onion seed, in this study, onion was used as the experimental material to clone the cDNA region of AcLOX (GenBank No. KX427168.1) by rapid amplification of cDNA ends (RACE) technology. Bioinformatics analysis was used to study its gene sequence characteristics; subcellular localization was used to study its expression site. qRT-PCR was used to identify the expression pattern of AcLOX in different parts of onion seeds at different stages. The results showed that AcLOX2 was 3 087 bp in length and contained a 2 727 bp ORF which encoding 908 amino acids. Subcellular localization revealed that AcLOX2 was located in mitochondria. qRT-PCR results indicated that AcLOX2 was expressed in all tissues measured, and its expression level was positive related to aging time. This experiment preliminarily confirmed that AcLOX2 played an important role in the aging process of onion seeds, and lays a foundation for studying the mechanism of short-lived mechanism of onion seeds.

Immune rejection is the main reason for limiting xenotransplantation. In this paper, α-1, 3-galactosyltransferase (GGTA1) and GGTA1/β1, 4 N-acetylgalactosaminyltransferase (β4GalNT2) gene knockout cloned pigs (Sus scrofa) were selected for donors of low-immunogenic xenotransplantation organ transplants. The expression of GGTA1/β4GalNT2 gene was identified by integrating peripheral blood mononuclear cells (PBMC) with fluorescein isothiocyanate-griffonia simplicifolia isolectin B4 (FITC-GSIB4) and fluorescein isothiocyanate Dolichos bliflorus agglutinin (FITC-DBA); the effect of GGTA1/β4GalNT2 double gene knockout on the health of Bama mini-pigs was assessed by blood changes in physiological and biochemical indicators; analysis of ideal receptor blood type was by detecting IgG binding of mixed PBMC of different genetically modified pigs and human serum of ABO blood group system after incubation; screening for low immunity primary donor genotype was bdye tecting IgG binding of mixed of PBMC of different genetically modified pigs and human (Homo sapiens) blood serum of AB blood group system after incubation. GSIB4 and DBA staining results showed that the expression of corresponding antigens on the surface of GGTA1/β-4GalNT2 gene knockout pig cells decreased; physiological and biochemical tests showed that GGTA1^-/- and GGTA1^-/-/β4GalNT2^+/- were compared with wild type (WT) Bama mini-pigs, genetically modified pigs had no significant differences in all indicators, GGTA1^-/-/β4GalNT2^-/- gene modified pigs had significantly reduced neutrophil counts, and significantly increased creatine kinase and creatinine; the results of incubation of serum of different blood types showed that compared with the amount of IgG binding of type A, B and O blood, the binding of IgG in type AB was the lowest, and it was the ideal recipient blood type; GGTA1^-/- and GGTA1/β-4GalNT2 knockout pigs, compared with Bama mini-pigs, had the lowest binding amount. The GGTA1/β-4GalNT2 knockout Bama mini-pigs were genetically stable, they also had reduced immunogenicity and immune rejection reactions with AB serum human serum were lowest. This study provides immunorejection data for future xenotransplantation clinical trials.

Genomic imprinting is an epigenetics phenomenon in mammals that leads to genes expressed from only one of the two parental copies. Imprinted genes play important roles in mammalian growth and development and affect the occurrence of diseases. Glutaminyl-peptide cyclotransferase (QPCT) has been found to be involved in post-translational modification of proteins and implicated in various mental disorders in human (Homo sapiens). Qpct is a placental-specific paternally imprinted gene in mouse (Mus musculus), but its imprinting status in cattle (Bos taurus) is still unknown. The aim of this study was to identify the imprinting status of QPCT gene in cattle and to determine whether DNA methylation would play a role in QPCT imprinting. The allele expression status of QPCT gene in bovine placenta and 6 somatic tissues (heart, liver, spleen, lung, kidney and brain) were analyzed by direct sequencing of reverse transcription PCR (RT-PCR) products based on single nucleotide polymorphisms (SNP). Results indicated that QPCT showed monoallelic expression in all the 6 tissues and placentas. QPCT showed maternal allele expression in bovine placentas by analyzing the genotypes of parental genomic DNA. The methylation status of CpG-enriched region in QPCT promoter were analyzed in bovine placentas and somatic tissues using bisulfite sequencing method, and no differentially methylated regions were detected in 55 CpG sites. The above results indicated that the DNA methylation modification in the promoter region of QPCT gene might not be involved in regulating the expression of QPCT gene in cattle, which would provide valuable information for further study of the function of QPCT gene.

3β-hydroxysteroid dehydrogenase 1 (HSD3B1) is a key member of the steroid hormone family and is closely related to animal reproductive traits. In order to clone the HSD3B1 gene of the Qianbei Ma goat (Capra hircus), construct a eukaryotic expression vector, and explore its effect on the expression of genes, in this study, Qianbei Ma goats ewes were selected to collect ovarian tissue for ovarian granulosa cell culture and extract the total RNA in the tissue for reverse transcription into cDNA. The full-length sequence of HSD3B1 (GenBank No. NM_001135932.1) gene CDS region of Qianbei Ma Goats was cloned and analyzed by bioinformatics. Meanwhile, eukaryotic expression vector of HSD3B1 gene was constructed and transfected into ovarian granulosa cells. mRNA expression levels of HSD3B1, bone morphogenetic protein 15 (BMP15), growth differentiation factor 9 (GDF9), and bone morphogenetic protein receptor type 1B (BMPR-1B) genes were detected by qRT-PCR. The results of protein prediction analysis showed that the molecular formula of HSD3B1 was C₁₉₄₀H₃₀₀₂N₅₀₆O₅₄₃S₁₅, and the relative molecular weight was 42.583 14 kD, isoelectric point 7.96; The genetic tree analysis showed that the genetic distance between the HSD3B1 gene and the cattle (Bos taurus) was the closest. The results of qRT-PCR showed that the expression of HSD3B1 and GDF9 gene in the transfection experimental group were significantly higher than those in the blank group (P<0.01), and the mRNA expression level of BMP15 gene was significantly higher than that of the control group (P<0.05). The results showed that the overexpression of HSD3B1 gene could promote the expression of GDF9 and BMP15. In this study, the pEGFP-N3-HSD3B1 eukaryotic expression vector was successfully constructed to detect the effect of HSD3B1 gene on the expression of genes related to lambing traits of Qianbei Ma goats, and provide basic data for further research on the effect of HSD3B1 gene on lambing traits of Qianbei Ma Goats..

Aurora kinase A (AURKA), one of the protein kinases, regulates mitotic protein kinase in cell cycle. Previous studies showed AURKA might be involved in regulating the development and maturation of germ cells. AURKA is a target gene of androgen receptor (AR) and dihydrotestosterone (DHT) to promote sperm maturation by combining with intracellular AR, which is of great significance for the study of animal reproduction and diseases, but the function of AURKA remains unclear. In the present study, the expression level and localization of AURKA in hypothalamus, pituitary and testis in male mice with 8 weeks were analyzed using semi-quantitative PCR, Western blot (WB), hematoxylin-eosin staining (HE) and immunohistochemistry (IHC). The designed specific knock down sequence of AURKA was transfected into the primary cultured sertoli cells. The expressed variation of AURKA and 5α-reductase type 1 (SRD5A1) mRNA and proteins was detected using the real-time fluorescent quantitative PCR (qRT-PCR) and WB after transfected for 24 h. The concentration of DHT in sertoli cells treated with or without AURKA-siRNA was determined using the enzyme-linked immunosorbent assay (ELISA). The results showed that AURKA mRNA and protein were expressed in the hypothalamic-pituitary-gonadal axis (HPGA) tissues of male mice. Compared to the expression of AURKA in hypothalamus and pituitary tissues, the highest expression level of AURKA mRNA and protein presented in testis (P<0.01). The strongest immuno-positive protein of AURKA showed in sertoli cells and sperm. The expression level of AURKA mRNA and protein was down-regulated (P<0.05), whereas the expression level of SRD5A1 mRNA and protein was significantly up-regulated (P<0.05) and the the concentration of DHT increased (P<0.05) in adult mice sertoli cells 24 h after the interference with AURKA-siRNA. The results suggested the expression of SRD5A1 and DHT synthesis were negatively regulated by AURKA. This data provide some theoretical basis for understanding the function of AURKA in the reproductive processes of male animals.

As an important economic trait and breed characteristic of duck (Anas platyrhynchos), plumage color has important research significance, but its genetic mechanism is still unclear. In order to study the differences in the transcriptome of white and black feather bulbs and offer new information related to gene expression profiles in black and white feather bulbs in ducks, 4 F₁ white-black plumage crossbred ducks of 'Liancheng white duck' and 'Cherry Valley duck' were selected to analyze the transcriptome characteristics of white and black feather bulbs in the same individual in this study. 457 208 376 raw cleans were obtained in 8 samples. After removing sequencing adaptors and the low-quality reads, 449 067 116 clean reads were obtained, accounting for 98.23% of the raw reads. The total base number of which was 67.36 Gb, and each sample obtained bases above 7.4 Gb, with the average GC content of 8 samples being 52%, the Q20 base percentage at 96.73% and above, and average 73.94% being successfully mapped to the duck genome. 70.85% of the mapped reads were uniquely aligned to the duck genome. The expression genes were screened using FPKM (expected number of fragments per kilobase of transcript sequence per millions base pairs sequenced)≧1 as the threshold. It was found that 14 296 genes were expressed in one sample at least. 231 significantly differentially expressed genes were identified (P≦0.05, |log₂Fold Change|≧1) between white and black feather bulbs, including 56 down-regulated and 175 up-regulated in black feather bulbs when compared with the white feather bulbs. Several genes related to melanogenesis were found in these differentially expressed genes, such as tyrosinase-related protein1 (TYRP1), tyrosinase (TYR), stem cell factor receptor (KIT), melanophilin (MLPH), dopachrome tautomerase (DCT/TYRP2), melan-A (MLANA), transient receptor potential cation channel subfamily M member 1 (TRPM1), oculocutaneous albinism typeⅡ(OCA2), SOX gene family member 10 (SOX10) and so on, and the expression of these genes in black feather bulbs was significantly higher than that in white feather bulbs. With Gene Ontology (GO) annotation, 19 differentially expressed genes were significantly enriched in 8 GO terms, including G-protein coupled receptor signaling pathway, transmembrane receptor activity, molecular transducer activity, and so on. Analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway showed that 3 pathways were enriched significantly, including neuroactive ligand-receptor interaction signaling pathway, tyrosine metabolism signaling pathway and melanogenesis signaling pathway. The melanogenesis and tyrosine metabolism signaling pathway played an important role in the formation of duck plumage color, and 5 differentially expressed genes including TYRP1, TYR, KIT, DCT etc. were involved in melanogenesis signaling pathway; TYRP1, TYR, DCT were involved in tyrosine metabolism signaling pathway. qRT-PCR was performed to verify the accuracy of results from RNA-seq and carried out for 6 randomly selected genes, including TYR, TYRP1, MLANA, synaptoporin (SYNPR), OCA2 and solute carrier family 38 member 11 (SLC38A11). Results of qRT-PCR were consistent with the data of RNA-seq. The results showed that down-regulation of TYRP1, TYR, KIT, DCT, SLC45A2, TRPM1, OCA2, SOX10, MLANA, MLPH and other genes in feather bulbs might be one of the causes of white plumage in the duck, revealed that tyrosine metabolism and melanogenesis signaling pathway played an important role in the formation of duck plumage color phenotype. This study would offer new information related to gene expression profiles in black and white feather bulbs in the duck, and provide basic materials for exploring the genetic mechanism and molecular marker breeding of duck feather color traits.

In China, the pigeon (Columba livia) production ranks fourth after the production of chickens (Gallus gallus), ducks (Anas platyrhynchos), and geese (Anser cygnoides orientalis). The pigeon has many valuable characters, such as high nutritional value and fast growth rate. However, little information is available about the genetic diversity of the pigeons used for agricultural production. The study on the genetic diversity and variety identification of the pigeon is of great significance for genetic conservation and inbreeding programs. The sequence of the mitochondrial cytochrome oxidase subunit Ⅰ (COI) has proven to be useful for the study of the population genetics of domestic animals. For this study, the complete COI gene sequences of 60 pigeons from three breeds (White Feather King, Thaxon, and Silver King) were analyzed. Other COI gene sequences were downloaded from GenBank, including pigeon species from other countries including the rock pigeon (Columba rupestris). No insertions or deletions were found in the COI gene. The sequence read length was 1551 bp, and encoded 517 amino acids. The average nucleotide composition was 25.6% T, 32.1% A, 26.1% C, and 16.2% G, and the content of AT (51.7%) was significantly higher than that of GC (48.3%), indicating slight base bias. Seven polymorphic sites were identified, representing 0.45% of the total analyzed sites. Six haplotypes were identified in 60 pigeons from the 3 investigated populations. The most common haplotype was Hap1, which was present in 26 out the 60 birds and had a frequency of 43.3%. The next-widespread haplotype was Hap 2 with a frequency of 25.0%. Two haplotypes were population-specific: Hap 4 for the White Feather King, Hap 5 for the Silver King pigeon. The overall haplotype diversity was 0.726±0.038. The highest haplotype diversity was found in the Silver King pigeon (0.716±0.069) whereas the lowest diversity was found in the White Feather King pigeon (0.674±0.049). The nucleotide diversity was 0.000 98±0.000 44 across the three investigated breeds, ranging from 0.000 54±0.000 27 in the White Feather King to 0.001 16±0.000 45 in the Silver King pigeon. The overall mean number of nucleotide differences was 1.518. The highest haplotype diversity was found in the Silver King Pigeon (1.795) and the lowest diversity was found in the White Feather King Pigeon (0.842). Based on the COI gene sequences of the 6 haplotypes obtained in this study, and other COI gene sequences, downloaded from GenBank, the rock pigeon (C. rupestris) was used as outgroup, and the molecular phylogenetic tree was constructed by the neighbor-joining method, based on the Kimura-2 parameter mode. Phylogenetic analysis showed that pigeons clustered into 2 cades (A and B). Only one haplotype (Hap 3) was distributed in Clade B, while all other sequences were clustered in Clade A. The median-joining network was constructed using the six haplotypes identified in this study, and the obtained haplotype network showed a star-like phylogeny. Most of the haplotypes were closely related to the common central haplotype (Hap 1), thus suggesting population expansion. The results of this study indicated that the COI gene sequence of pigeons was relatively conservative and contained few mutation sites. This was less effective for the identification of different pigeon breeds. Consequently, there was a high need to incorporate multiple molecular markers such as single nucleotide polymorphism (SNP), simple sequence repeat (SSR), and copy number variations (CNV). The COI gene could be used as a candidate molecular marker to investigate the genetic diversity and origin of pigeon species, and can thus serve as a factual and comparative basis for similar studies in the future.

BLM (bloom helicase) plays an important role in cell proliferation and growth. YAP (yes associated protein) and TAZ (transcriptional coactivator with PDZ-binding motif) porteins, as transcriptional coactivators and key components downstream of the mammalian Hippo signal pathway, are involved in the regulation of cell proliferation, apoptosis, migration and other physiological processes.And the long non-coding RNA (LncRNA) is very critical for regulating gene expression in cell growth and proliferation. To explore LncRNAs effect on gene expression of BLM, YAP, TAZ and screen BLM helicase related LncRNAs, fluorescence quantitative PCR technique had been used to detect the expression of selected LncRNA in 4 kinds of cell lines (22RV-1, PC3, LNCap, WPMY-1). Construction of LncRNA over-expression vector had been transfected in PC3 cells and WPMY-1 to detect the impact on BLM, YAP and TAZ gene after transfection LncRNA 24 and 48 h by using the fluorescent quantitative PCR technique. The NONHSAT184603.1 (abbreviated as N183) (GenBank No. m121218_085412_00126_c100) was successfully screened as candicate. The fluorescence quantitative PCR results showed that the expression of N183 in 22RV-1 and LNCap cells was significantly lower than that in normal prostate epithelial cell line WPMY-1 (P<0.01), and the expression in PC3 cells was significantly higher than that in WPMY-1 (P<0.01). After transfection with N183 over-expression vector, compared with the control group, the expression of BLM and TAZ were significantly down-regulated 24 h after transfection (P<0.01), and the expression of BLM and YAP were significantly up-regulated 48 h after transfection (P<0.01). BLM expression was significantly down-regulated (P<0.05), but there were no significant difference in YAP and TAZ expression in PC3 cells 48 h after transfection with N183 over-expression vector (P>0.05).The above results indicated that N183 could inhibit BLM gene expression in prostate cancer cells and normal cells but had different effect no YAP and TAZ genes, which made a contribution of basic data for anti-cancer researches by targeting BLM helicase.

Salpingitis and oviduct cyst of laying hens (Gallus?gallus domesticus) are serious diseases harmful to poultry industry. Gallibacterium anatis is one of the common pathogens. At present, Lactic acid bacteria (LAB) are widely used in the development of live vector vaccine as food-grade expression vector. Therefore, Lactobacillus casei CECT5276 (L. casei 5276) was used as a live vector for presenting antigens in this study. With L. casei 5276 as the host bacterium and G. anatis flfA gene as a target, a shuttle expression vector pMG36e were used to obtain recombinant plasmid pMG36e-flfA. The recombinant plasmid was transferred into L. casei by electroporation, then the expression of flfA protein and the growth status, stability, retention and tolerance of recombinant bacteria were detected and analyzed, and the immune effect of recombinant bacteria was preliminarily evaluated. The analyses of SDS-PAGE and Western blot showed that the recombinant L. casei successfully expressed fusion protein with a molecular weight of about 20.3 kD, which could react with Rabbit anti-G. anatis positive serum and had reactogenicity. The analysis of growth status and stability showed that the target gene insertion had no effect on the growth of L. casei and could be stably inherited in the host bacteria; it had good tolerance in simulated intestinal juice, gastric juice and high salt environment. On the 0, 14, 28, 42 d after immunization, the specific IgG and SIgA antibody levels in serum and intestinal mucus were higher than those in empty vector group and PBS control group (P<0.05). The results of animal challenge test showed that all chickens in 3 groups did not die after intraperitoneal injection of G. anatis PDS-RZ-1-SLG strain, but the isolation rate of G. anatis from chicken organs immunized with recombinant bacteria was lower than that of empty vector group and PBS group, suggesting that recombinant bacteria had certain immune protective effect on chickens. This study could provide a reference for further study of oral vaccine against G. anatis.

The rice (Oryza sativa) monogenic line of 'IRBL9-W' (containing resistance gene (Pi9)) was resistant to most of the Magnaporthe grisea isolates collected in Jilin province, indicating that the isolates contain the avirulence gene (AVR) widely spread in Jilin Providence. To investigate the gene expression profiles of the rice during the early interaction between Pi9 and avirulence gene (AVR-Pi9), the rice monogenic line of 'IRBL9-W' (Pi9) inoculated with Magnaporthe grisea isolate C2 (AVR-Pi9) in this study. The RNA-seq technology was used to analyze the expression profiles of the inoculated and uninoculated rice leaves of 'IRBL9-W' (Pi9) at 36 h after inoculation and a total of 2 311 genes were found to express differently in the two samples. The analyses of GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways showed that 3 of the 4 leucine zipper protein genes were up-regulated, 1 zinc finger protein gene was up-regulated and all the WRKY protein genes were down-regulated among transcriptional regulator genes, 2 chitinase genes were up-regulated among the10 chitinase genes and all the two β-1,3 glucanase genes were down-regulated among the fungal cell wall degrading enzyme genes, 1 pathogenesis-related protein 1 gene (PR1) was up-regulated and the other genes were down-regulated in the plant-pathogen interaction pathway, all of the differentially expressed genes were up-regulated in the flavonoid biosynthesis pathway, and 9 genes were up-regulated and 1 gene was down-regulated in the phenylpropanoid biosynthesis pathway. The differentially expressed gene products in the 3 pathways were involved in getting the reactive oxygen species, the hypersensitive response, and the antioxidant process. The functions of the compounds relating to these gene products were antioxidation, antifungal action, and lignification. This study would provide a theory to further understand the mechanism of rice blast resistance.

Bucket worms are larva of longhorn beetles Apriona germari and A. swainsoni living in the trunk of Caesalpinia decapetala (Leguminosae). These insects are considered of high nutritional and medicinal value, and therefore widely consumed. The purpose of this study was to characterize the microflora of fresh edible bucket worms by the methods of high-throughput sequencing and traditional culture, and further evaluate the food safety based on relative abundance of Enterobacteriaceae, Saccharomycete, molds, and lactic acid bacteria (Lactobacillales) according to GB4789.1-2016 and ISO standards, and culture-dependent method was used to analyze thermoduric bacteria. The results showed that there were no significant differences between the microbial community compositions of 2 bucket worm species. The bacterial community of A. germari was mainly composed of Proteobacteria, Oxyphotobacteria and Bacteroides, while A. swainsoni was mainly composed of Proteobacteria, Firmicutes and Actinobacteria. Ascomycota and Basidiomycota were dominant fungal phyla in 2 bucket worm species, in which Ascomycetes were the absolutely dominant group. The analysis of safety-related indicators showed that relative abundance of Enterobacteriaceae in A. germari and A. swainsoni accounted for 60.28% and 39.82%, while Lactobacillus accounted for 1.24% and 3.65% respectively in bacteria; the molds accounted for 40.26% and 6.74% in fungal, while the yeast accounted for 59.74% and 93.26% respectively. Among yeast, 21 genera were found common in bucket worms, which were considered to be toxic or harmful to human beings (Homo sapien). In addition, 4 genera of potentially poisonous thermoduric bacteria including Staphylococcus, Streptomyces, Acinetobacter, and Pseudomonas were identified in 2 insect species, the colony number was significantly reduced after treatment at 50 ℃, 60 ℃ and 70 ℃, and almost inactivated completely at 80 ℃. Overall, the results of this study indicated that there were a variety of potential spoilage bacteria and food pathogens in the bucket worms, and the food safety risk could be eliminated or minimized through processing steps such as a shock heat treatment. The microbial safety evaluation based on high-throughput sequencing in present study could provide reference for hygiene criteria of edible insects.

Synthetic biology brings together engineering and the life sciences in order to design and construct new biological parts, devices and systems that do not currently exist in the natural world or to tweak the designs of existing biological systems. Compared with traditional transgenic technology, synthetic biology uses new technologies in genome design, synthesis and more effective molecular tools, which can develop new varieties and new technology applications more efficiently and extensively. Because synthetic biology has the characteristics of strong targeting, simple operation, short application period, and high sensitivity, the use of synthetic biology to replace traditional agricultural technology and biotechnology can bring great potential to the transformation of agricultural production. This article reviews the research progress of functional microorganisms based on synthetic biology in soil monitoring and repair, plant growth promotion, plant pathogen detection, agricultural product safety detection, plant natural product production, etc., and clarify the potential problems of synthetic biology used in planting production at this stage, analyze and propose solutions at the database level, technical level, security level, and regulatory level.

Diarrhea of piglets in large-scale farms is often caused by different viruses, which have similar clinical symptoms in winter and spring. In order to distinguish the pathogens in clinical samples of viral diarrhea, it is urgent to establish a rapid and simultaneous detection technology for viral nucleic acid of six viral diarrhea diseases. In this study, the probes and pre-amplification primers for multiple restriction probe amplification (MLPA) were designed countering for the conserved regions of nucleic acids of Porcine epidemic diarrhea virus (PEDV) S gene, Transmissible gastroenteritis virus (TGEV) N gene, Porcine delta coronavirus (PDCoV) N gene, Porcine Bocavirus (pBCaV) NS1 gene, Porcine norovirus (pNov) RdRp gene, Porcine rotavirus (PRV) NSP1 gene. The method of detecting nucleic acid of 6 diseases was established using MLPA and capillary electrophoresis. The results showed that the detection of the 6 mixed templates by a single probe showed a high specificity. When the total concentration of the mixed probes were 1.33 nmo/L, and the specific amplified bands of various diseases were the same as expected, with the products of pBCaV (102 bp), pNov (110 bp), PDCoV (117 bp), PEDV (124 bp), pRV(131 bp) and TGEV (138 bp). This method had no cross reaction with other clinical diseases about Porcine reproductive and respiratory syndrome virus (PRRSV), Classic swine fever virus (CSFV), Pseudorabies virus (PRV) and Porcine circovirus type 2 (PCV2). The minimum limit values for detecting nucleic acid of pBCaV, pNoV, PDCoV, PEDV, pRV, TGEV were 7.58×10¹, 7.56×10⁰, 7.54×10⁰, 7.53×10⁰, 7.50×10¹ and 7.49×10⁰ copies/µL, respectively. The repeatability between and within groups was well. The results of 67 clinical and simulated samples by this method showed that PEDV were 100% in accordance with the virus isolation method, pBCaV, pNoV, PRV, TGEV and PDCoV were 100% in accordance with simulated virus. This study provides a new technology for detecting 6 clinical diarrhea pathogens simultaneously, and for the rapid response of clinical disease prevention and control.