Biology method is one of the most economic and effective ways for the treatment of aquaculture waste at present. Nitrifying bacteria plays an important role in the process of biological nitrogen removal. A highly efficient heterotrophic nitrifying bacterium P5-2 was isolated from the water of tilapia (Oreochromis niloticus) recirculation aquaculture systems through enrichment culture and plate purification. The strain was identified as Bacillus megaterium by morphological observation, physiological and biochemical reaction identification and 16S rRNA gene sequence analysis. The results of Gram staining showed that the strain was Gram-positive. The result of determination of extracellular enzyme showed that starch and casein could be utilized by the strain. When temperature was 30 °C, salinity was 10‰, and pH was 7, the strain grew fastest, and first reached the logarithmic growth period and the stable period. The results of nitrification experiments showed that environmental factors such as carbon source, carbon-nitrogen ratio and dissolved oxygen concentration could influence its rate of ammonia nitrogen removal. When sodium citrate was used as carbon source, carbon-nitrogen ratio was 15, pH was 7, and rotational speed was 180 r/min, the highest ammonia nitrogen removal rate of 99.07% was achieved after 24 h. A highly efficient heterotrophic nitrifying bacterium P5-2 was isolated in this study. Its nitrification performance was explored, which provides critical support for its application in aquaculture waste water treatment.

AKT1 (Arabidopsis potassium transport 1) is an important potassium channel protein gene involved in plant potassium absorption and transport. In order to study the response and expression pattern of PbAKT1 gene in Pyrus betulifolia to potassium, this study adopted K⁺ at different concentrations (0.1 mmol/L and 3 mmol/L K⁺) to treat pear seedlings, cloned PbAKT1 gene and analyzed the tissue specificity of its encoded protein. Results showed that the PbAKT1 gene (GenBank No. MN150549) cDNA was 2 646 bp in length, encoding 881 amino acids. The gene had a transmembrane structure with a typical S1~S6 of the Shaker family and a loop ring. The homology analysis of amino acid sequence indicated that there was a close phylogenetic relationship between PbAKT1 gene and MdAKT1 gene (Malus domestica). Subcellular localization showed that the gene mainly presented in the cell membrane and was a hydrophilic stable protein. PbAKT1 gene was found no tissue specificity in roots, stems and leaves of pear seedlings and its expression obviously responded to low potassium stress. It was indicated that the expression of PbAKT1 was induced by potassium supply and might be involved in K⁺ absorption and transport of pear trees. It provides basic information for further research of the molecular mechanism to low potassium stress in pear.

The calcineurin B-like protein (CBL), which is a unique group of calcium sensors in plant, play a key role in stress response. Sugarcane (Saccharum spp.) is an important crop for sugar and bio-energy in China. According to the datasets which are transcription sequencing of sugarcane genes under abiotic stress, a calcineurin B-like gene named SsCBL4 (GenBank No. KY674 987.1), was cloned from root in sugarcane under low potassium stress by RT-PCR (reverse transcription-PCR). SsCBL4 contained a 636 bp complete open reading frame that encodes 211 amino acids. Its molecular weight was 23. 97 903 kD. And the isoelectric point of SsCBL4 was 4.88. The deduced polypeptide of SsCBL2 had 2 transmembrane domains which could bind calcium ions. The SsCBL4 protein contain 3 EF-hand domains (elongation factor hand) that are the typical domain of CBL family, a conserved N-myristoylation motif and FPSF motif which interacted with CBL-interacting protein kinase (CIPK) kinases. The homology analysis indicated that SsCBL4 has high homology to the CBL4 from Sorghum bicolor, Zea mays, Oryza sativa and Triticum aestivum. The changes in expression of SsCBL4 under low nitrogen, low phosphorus, low potassium, drought or salt stresses were detected by qPCR analysis. The expressions of SsCBL4 were commonly up-regulated under low potassium, drought or salt stresses. Under low nitrogen or low phosphorus stress, the fluctuations in SsCBL4 gene expression levels were observed, with the highest level at 96 h, followed by a significant decrease. Under low potassium stress from 8 to 48 h, the expression of SsCBL4 gradually decreased. SsCBL4 was induced to express at relatively high levels at 8, 24 and 96 h after drought treatment, and reached the highest level at 96 h, which was 20.5-fold as high as that of control. Under salt stress from 24 h to 96 h, the expression of SsCBL4 gradually increased, and reached the highest inducible expression level at 96 h, which was 9.1-fold as high as control. These results suggest that SsCBL4 gene may play an important role in different stresses. This study provides a reference for further study of SsCBL4 stress resistance and related molecular mechanisms.

Coix (Coix lachryma-jobi) is a crop of food and medicine, and has very high nutritional and medicinal value. But its genetic resources are scarce. Uniconazole (S3307) is plant growth retardant belong to triazole category, which can inhibit the biosynthesis of endogenous gibberellin and has obvious effects on plant growth and stress resistance. In order to obtain and dig the genetic data and function of Coix lachryma-jobi and explore the alleviating effect of S3307 on low temperature stress of coix seedlings, 'Yiliao 5' was used as experimental material, Li-6400XTR photosynthetic apparatus and transcriptome sequencing technology were used to explore the effects of S3307 on photosynthetic characteristics and transcription factor gene expression of coix seedlings under low temperature stress. The results showed that addition of 5 mg/L S3307 could significantly increase the chlorophyll contents of coix leaves under low-temperature. Meanwhile, the net photosynthetic rate, transpiration rate and stomatal conductivity enhanced 138.1%, 59.59% and 50.15%, respectively. Through screening analysis and Swiss-Prot database annotation, 4 differentially expressed genes (DEGs) of WRKY transcription factor, 6 DEGs of bZIP (basic leucine zipper), 9 DEGs of bHLH (basic helix-loop-helix) and 6 DEGs of AP2/ERF (APETALA2/ethylene-responsive factor) were obtained. The qRT-PCR was used to analyze the expression of 10 DEGs, and the changing trend was consistent with the results of transcriptome sequencing. The results of this study will help to reveal the mechanism of S3307 on photosynthetic characteristics and transcription factor expression characteristics of coix under low temperature stress, and provide a reference for further research on transcription factor function in response to low temperature stress.

ω-3 polyunsaturated fatty acid (PUFA) dehydrogenase, the expression product of Fat1 in Caenorhabditis elegans, can convert ω-6 PUFAs to ω-3 PUFAs. Mammals cannot synthesize ω-3 PUFAs due to lack of Fat1 gene. ω -3 PUFAs can only be obtained through the diet to ensure the balance of ω-6/ ω-3 and body's health. In order to obtain the transgenic pig (Sus scrofa) with stable integration and expression of Fat1, experiments on transgenic technology at the cellular level were performed by zinc finger nuclease (ZFN)-mediated multi-site targeting technology. Expression vectors pcDNA3.1-NLS-ZFN-R and pcDNA3.1-NLS-ZFN-L, which were used to target to internal transcribed spacer-1 (ITS1) sequence in Luchuan pig rDNA gene, were constructed. Secondly, primary kidney fibroblast (PKF) cells were successfully isolated from kidney. After 7 d of co-transfection of targeting vector and ZFN vectors into PKF cells, the genomic DNA was extracted and the integration of exogenous genes were detected by PCR. The results showed that Fat1 gene was successfully integrated into the genome of Luchuan pig PKF cells, but not in a site-specific way. This study provides a theoretical basis for further obtaining transgenic pigs.

Congjiang Xiang pig (Sus scrofa) has the characters of short and small body and slow growth rate. For preliminarial exploreation on Congjiang Xiang pig's characters and providing basic data on the molecular mechanism of growth and development of miniature pigs, insulin-like growth factor 1 and 2 (IGF-1 and IGF-2) genes had been analysed by bioinformatics methods as well as their expression level of mRNA in various organs and tiusses from Congjiang Xiang pig and Norther Yorkshinre had been detected. In the current, the coding sequences (CDS) of IGF-1 (GenBank No. NM_214256.1) and IGF-2 (GenBank No. NM_213883.2) genes were had successfully cloned by using Congjiang Xiang pig and Norther Yorkshinre with same feeding condictions and ages and had predicted and dissected on the structures and functions of IGF-1 and IGF-2. The qRT-PCR had been applied to detect mRNA levels of IGF-1 and IGF-2 of Congjiang Xiang pig and North yorkshire in the heart, liver, spleen, lung, kidney, large intestine, small intestine, fat, and longissimus 9 organs or tissues. The results showed that the CDS of IGF-1 and IGF-2 of Congjiang Xiang pig were 435 and 577 bp respectively, which were no difference with CDS sequence of Sus scrofa in GenBank. By sequence prediction, the relative molecular mass, theoretical isoelectric point, and unstable coefficient of IGF-1 and IGF-2 protein resplectively were 14.390 44 and 20.312 5 kD, 9.34 and 10.00, 65.58 and 58.64. Comparing other 12 species by genetic evolution analysis showed that Congjiang Xiang pig was the closest with S. scrofa. The qRT-PCR results displayed that IGF-1 gene had the obviously highest expression level in lung, large intestine, and small intestine of Congjiang Xiang pig (P<0.05) as well as IGF-1 gene had the very significantly highest expression level in longissimus of North Yorkshine (P<0.01) by comparing other organs or tissues. The IGF-2 had obviously higher expression level in liver and lung than other organs and tissues in Congjiang Xiang pig but longissimus and heart of North yorkshine had very significantly higher expression level of IGF-2 gene than other organs or tissues (P<0.01). Our research uncovered the expression difference of IGF-1 and IGF-2 genes between Congjiang Xiang pig and North Yorkshine, which provided basic data for further exploration of the molecular mechanism of growth and development of miniature pigs.

Tribbles pseudokinase 3 (TRIB3) is a serine/threonine kinase-like protein, which is involved in insulin signal pathway as a negative regulator of lipid metabolism. The present study was aimed to obtain the TRIB3 gene sequence of Xinong Saanen dairy goats (Capra hircus), elucidate its structure and investigate its function on lipid metabolism through RNA interference of TRIB3 gene in goat mammary epithelial cells. Taken mammary gland from Xinong Saanen dairy goats as experimental materials, the sequence of TRIB3 gene was cloned by RT-PCR method, bioinformatics and tissue expression analyses were conducted. The results showed that the complete CDS region of TRIB3 gene was 1 074 bp (GenBank No. MK158243), encoding 357 amino acids. The TRIB3 protein had the molecular weight of 39.6 ku, the isoelectric point of 7.8, and was an unstable protein. The TRIB3 protein had 36 phosphorylation sites without signal peptides and transmembrane spiral structure. Homology analysis showed that sequence homologies of Xinong Saanen dairy goat with sheep (Ovis aries), cattle (Bos taurus) and mouse (Mus musculus) were 99%, 96% and 71%, respectively, and the sheep was the most closest species with dairy goat. Tissue expression analysis revealed that the highest expression of TRIB3 was in mammary gland, followed by adipose tissue. In addition, siRNA of TRIB3 gene were designed and transferred into goat mammary epithelial cells, the results showed that mRNA level of peroxisome proliferator activated receptor γ (PPARG) and diacylglycerol acyltransferase (DGAT1) were up-regulated by interfering TRIB3. Above results suggest that TRIB3 might be involved in the regulation of lipid metabolism in goat mammary epithelial cells as a negative factor. The present study provides basic data for further investigation of molecular mechanism of TRIB3 on milk fat metabolism in dairy goats.

Most of the sheep (Ovis aries) are seasonal estrus animal, which severely restricts the reproductive efficiency of sheep, and retinoid X receptors (RXRs) have received attention as seasonal estrus candidate genes. In order to investigate the differences in the expression of seasonal estrous candidate genes retinoid X receptors (RXRA, RXRB and RXRG) in sheep under different reproductive conditions, qRT-PCR was used to detect these 3 genes for the relative expression of 10 tissues about fallopian tube, brain, cerebellum, hypothalamus, pineal body, pituitary, ovary, fallopian tube, uterus, kidney and adrenal gland, in the follicular and luteal phases of Small Tail Han sheep (perennial estrus) ewes and the long day and short day conditions of the Sunite sheep (seasonal estrus) ewes. The Sequenom MassARRAY^® SNP technique was used to detect the 2 SNPs of RXRA gene in perennial estrus sheep breeds (Small Tail Han sheep, Hu sheep and Cele black sheep) and seasonal estrus sheep breeds (Tan sheep, Sunite sheep and Tibetan sheep). The results showed that RXRA, RXRB and RXRG genes were widely expressed in the tissues of the 2 sheep breeds. The expression of RXRA gene in the pineal body of Sunite sheep in short day was significantly higher than that in long day (P<0.05), and the expression of follicular phase in the ovary of Small Tail Han sheep was extremely significant higher than the luteal phase (P<0.01), but the expression of follicular phase in the uterus was extremely significant lower than that in the luteal phase (P<0.01). The expression of RXRB gene in the pituitary of Sunite sheep in short day was significantly higher than in long day (P<0.05). The expression of follicular phase was extremely significant higher than that of luteal phase in the ovary of Small Tail Han sheep (P<0.01). RXRG gene was highly expressed in the hypothalamus, pituitary and pineal body of Sunite, and the expression level in pineal body in long day was extremely significant higher than in short day (P<0.01). The expression level in the uterus of the Small Tail Han sheep was extremely significant lower in the follicular phase than in the luteal phase (P<0.01). From genotyping by MassARRAY time-of-flight mass spectrometry, the study found the genotype frequencies and allele frequencies of the RXRA gene g.1699247 G>A and g. 1699276 G>A were distributed in the Small Tail Han sheep in accordance with the Hardy-Weinberg equilibrium, and were significantly different between seasonal and perennial estrus sheep (P>0.05). The above results implied that RXRs genes expression might be related to reproduction of sheep. These genes might be involved in the regulation of upstream genes of sheep seasonal estrus. The results of this study helps to understand the regulatory role of RXRs in seasonal estrus and estrus cycle transition.

Variable splicing enables a single gene to produce multiple mRNAs, thereby enhancing protein diversity, which is closely related to the growth and development of organisms. Among the abundant horse (Equus caballus) breed resources in China, Debao pony and Mongolian horse have obvious height differences. In this study, 137 449 alternative splicing events were detected by RNA-seq and bioinformatics analysis using hypophysis tissue which closely related to horse size. There were 4 main types of alternative splicing including intron retention (IntronR), alternative 3' splice site (A3SS), alternative 5' splice site (A5SS) and exon skipping (ES). In different horse breeds and different development durations, 37 genes presented differential alternative splicing, including 23 differentially alternative splicing genes between immature Debao pony and immature Mongolian horse, 1 differentially alternative splicing gene between adult Debao pony and adult Mongolian horse, 5 differentially alternative splicing genes between immature Debao pony and adult Debao pony, and 8 differentially alternative splicing genes between immature Mongolian horse and adult Mongolian horse. In alternative splicing types of hypophysis, cassette exon splicing of WDR35 gene and ES splicing of OSBPL6 gene in immature Debao pony were identified as important factors affecting dwarfism of Debao pony. Moreover, in the developing hypophysis of Debao pony and Mongolian horse, 13 genes including CPEB1, TCF12, CCDC28B, KIT, FAM129A, PTPRF, EPHA5, OGDH, USP28, EMID1, ANKHD1, KIDINS220 and INF2 were selected as candidate genes targeting dwarf trait by comparing differential alternative splicing types of immature and adult horses. Above dwarf-related genes provide basic information for further molecular breeding targeting horse height.

Tumor necrosis factor α (TNF-α) is a pro-inflammatory Th1 cytokine that is involved in the regulation of mammalian ovulation, embryonic development and implantation. The aim of this study was to investigate the role of TNF-α in the development of oocyte maturation and in vitro fertilization (IVF) embryos by detecting the expression of TNF-α in yak (Bos grunneins) oocyte maturation and early IVF embryo development. In this study, the cumulus-oocyte complex (COCs) were collected and matured in vitro and fertilized to obtain IVF embryos, and to evaluate the developmental ability of embryos at different stages of implantation. Methods of qRT-PCR, indirect immunofluorescence staining and Western blot were used to detect the expression and distribution of TNF-α in immature oocytes, mature oocytes and embryos at different stages. The results showed as follows: (1) The rates of embryos developed to stages of 2~4 cells, 5~8 cells, morulae were (61.59±0.31)%, (53.76±0.22)% and (26.88±0.32)%, respectively, and the blastocyst rate was (16.12±0.23)%. (2) qRT-PCR results showed that, TNF-α mRNA could be detected in immature COCs, mature COCs, morulae and blastocysts, the relative expression levels in the order of high to low was mature COCs, immature COCs, blastocysts and morulae. 2~4 cells and 5~8 cells embryos had no expression. (3) The result of indirect immunofluorescence and Western blot showed that the protein of TNF-α was mainly distributed in the cytoplasm of COCs and embryos, which was weaker in the nucleus. The protein of TNF-α could be detected in COCs, morula and blastocyst, the relative expression levels in the order of high to low was mature COCs, immature COCs, blastocysts and morulae, but 2~4 cells and 5~8 cells embryos had no expression. The results in this study indicated that TNF-α might play a role in the process of the maturation and senescence of the yak oocytes and might be involved in the process of blastocyst cell apoptosis and embryo implantation, which provides a theoretical basis for revealing the mechanism of TNF-α in yak reproduction.

As a crucial approach to detect candidate genes of important economic straits, GWAS (genome-wide association study) is used for identifying complex functional genes of straits on genomic level. To identify molecular markers and candidate genes associated with growth and carcass straits, a multibreed GWAS was performed based on growth and carcass trait phenotypes with the 600 K SNP array in Ninghai yellow chickens (Gallus gallus domesticus) and Guangxi yellow chickens. The statistics of the minimum, maximum, mean, standard deviation and coefficient of the growth traits and carcass traits of Ninghai yellow and Guangxi yellow chickens were made , which found that there were significant differences (P<0.01) in the body weight at 8 weeks old and 16 weeks old and slaughter rate in the 2 breeds . As shown in present study, the variation coefficients of abdominal fat weight and abdominal fat rate were the highest. Followed by body weight at 0, 8 and 16 weeks, slaughter rate, semi-eviscerated rate and eviscerated rate. The result of principal component analysis showed that there was no significant subgroup structure in the 2 chicken populations, respectively. The Quantile-Quantile (QQ) graph showed that the actual value of each character was basically consistent with the expected value. Finally, Manhattan plots of growth traits and carcass traits showed that the presence of loci on multiple chromosomes of 2 breeds of chickens was significantly correlated with growth traits and carcass traits, respectively. The SNPs that significantly associated with phenotypic traits were identified by a general linear model. Four SNPs (rs313150871, rs314661053, rs313314400 and rs314694861)on G. gallus domesticus (GGA)3, 5, 8, and 11, and 3 SNPs (rs313989383, rs317888074 and rs313384732) on GGA 2, 5, and 9 were found to be significantly associated with growth and carcass traits, respectively. Only one SNP (rs313384732) on GGA 5 at 11.80 Mb was significantly associated with 2 traits (semi-eviscerated weight, eviscerated weight). In particular, a region of GGA 5 between 8.98 and 11.80 Mb was associated with multiple traits (i.e. BW at hatching, semi-eviscerated weight, and eviscerated weight). Four proximal genes (adrenomedullin, ADM; ATP/GTP binding protein like 4, AGBL4; activating signal cointegrator 1 complex subunit3, ASCC3; wilms tumor 1 interacting protein, WTIP) for growth traits and 3 proximal genes (phosphatidylinositol-4-phosphate 3-kinase catalytic subunit type 2 alpha, PIK3C2A; mitogen-activated protein kinase kinase kinase 13, MAP3K13; and LOC101747362 (uncharacterized gene)) for carcass traits were detected. The findings would be helpful to reveal the genetic effects of traits and marker-assisted selection.

Spermine is one of the essential factors for cell proliferation and differentiation. The spermine in small intestine not only plays a role in regulating intestinal epithelial cell proliferation and apoptosis and promoting intestinal development, but also plays a role in regulating intestinal digestion and absorption. This experiment was conducted to study the effects of exogenous spermine on polyamine metabolism in female goose (Anser cygnoides) duodenum. Geese were fed spermine at 5 and 10 mg/kg (body weight). The expression of key genes related to polyamine metabolism and the content of polyamines in female geese duodenum were measured by qRT-PCR and high performance liquid chromatography. The gene expression levels of ornithine decarboxylase antizyme inhibitor (AZIN) 1, ornithine decarboxylase antizyme 1 (OAZ1), spermine synthase (SPMS), spermidine/spermine-N1-acetyltransferase (SSAT) and acetyl-polyamine oxidase (APAO) in the duodenum of female geese treated with spermine at 5 mg/kg were significantly higher than those in the control group (P<0.05), while the gene expression level of AZIN2 significantly decreased (P<0.05). The gene expression levels of AZIN1, OAZ1, ornithine decarboxylase (ODC), spermidine synthase (SPDS) and SSAT in the duodenum of female geese treated with spermine at 10 mg/kg were significantly higher than those in the control group (P<0.05). The contents of spermidine (65.08 vs 41.84 mg/kg) and spermine (255.79 vs 218.84 mg/kg) in the duodenum of female geese treated with spermine at 5 mg/kg were significantly higher than those in the control group (P<0.05). Spermidine content (56.20 vs 41.84 mg/kg) in the duodenum of female geese treated with spermine at 10 mg/kg was significantly higher than that in the control group (P<0.05), while spermine content (188.34 vs 218.84 mg/kg) significantly decreased (P<0.05). In summary, exogenous spermine affects the contents of spermidine and spermine in female goose duodenum through regulating the expression of key genes related to polyamine metabolism, while it had no significant effect on putrescine content. This study revealed the effects of exogenous spermine on the expression of key genes of polyamine metabolism and polyamine content in duodenum of female geese, which provides basic data for the study of spermine regulation on digestion and absorption of poultry, and provides a new way for the development of new functional feedstuffs.

The follicle stimulating hormone receptor (FSHR) is used to mediate the physiological functions of biomacromolecule follicle stimulating hormone, and has an important influence on the reproductive traits of animals. In order to analyze the relationship between single nucleotide polymorphisms (SNPs) of FSHR gene and egg production in White King Pigeon (Columba livia), this study selected 209 pigeons as the research subjects and used the direct sequencing of PCR products to screen the SNPs of exon 6, exon 9, exon 10 and their adjacent upstream and downstream sequence in FSHR gene. The results showed that 18 mutation sites were found in the 3 fragments detected and the amino acid sequences were changed accordingly. Among them, 3 mutation sites G67729A, A67801G, A67898G were found in exon 6 and its upstream and downstream sequences. 5 mutation sites C76462T, T76468C, G76527T, C76616T, T76651G were found in exon 9 and its upstream and downstream sequences. 10 mutation sites were found in exon 10, which were A82714G, C82930T, A83083C, G83098T, G83158A, C83200T, A83246C, T83395C, T83740C, C83766T. T76468C and C83766T had 2 genotypes AA, AB, and the rest had 3 genotypes: AA, AB, BB. A Hardy-Weinberg equilibrium test was performed on 18 loci and found to be in equilibrium except for the C76616T locus. Correlation analysis between polymorphic loci and average egg production showed that the homozygous BB genotype of A67801G locus was extremely significantly correlated with egg production (P<0.01), and the homozygous BB genotype of C76616T locus was significantly correlated with egg production (P<0.05). Therefore, the A67801G and C76616T loci in the FSHR gene could be utilized as candidate molecular marker sites for the selection and breeding of the egg-laying traits of White King Pigeons, which would provide a theoretical basis for marker-assisted breeding from the molecular level to better improve the breeding performance of the pigeon population and shorten the breeding years.

The follicle stimulating hormone receptor (FSHR) is used to mediate the physiological functions of biomacromolecule follicle stimulating hormone, and has an important influence on the reproductive traits of animals. In order to analyze the relationship between single nucleotide polymorphisms (SNPs) of FSHR gene and egg production in White King Pigeon (Columba livia), this study selected 209 pigeons as the research subjects and used the direct sequencing of PCR products to screen the SNPs of exon 6, exon 9, exon 10 and their adjacent upstream and downstream sequence in FSHR gene. The results showed that 18 mutation sites were found in the 3 fragments detected and the amino acid sequences were changed accordingly. Among them, 3 mutation sites G67729A, A67801G, A67898G were found in exon 6 and its upstream and downstream sequences. 5 mutation sites C76462T, T76468C, G76527T, C76616T, T76651G were found in exon 9 and its upstream and downstream sequences. 10 mutation sites were found in exon 10, which were A82714G, C82930T, A83083C, G83098T, G83158A, C83200T, A83246C, T83395C, T83740C, C83766T. T76468C and C83766T had 2 genotypes AA, AB, and the rest had 3 genotypes: AA, AB, BB. A Hardy-Weinberg equilibrium test was performed on 18 loci and found to be in equilibrium except for the C76616T locus. Correlation analysis between polymorphic loci and average egg production showed that the homozygous BB genotype of A67801G locus was extremely significantly correlated with egg production (P<0.01), and the homozygous BB genotype of C76616T locus was significantly correlated with egg production (P<0.05). Therefore, the A67801G and C76616T loci in the FSHR gene could be utilized as candidate molecular marker sites for the selection and breeding of the egg-laying traits of White King Pigeons, which would provide a theoretical basis for marker-assisted breeding from the molecular level to better improve the breeding performance of the pigeon population and shorten the breeding years.

Fungal diseases of tea are one of the important causes of tea yield reduction. As environmentally friendly antimicrobial materials, nano-materials can be used to inhibit tea fungal diseases. In this study, nano-Mg(OH)₂ was synthesized by coprecipitation method, and the inhibitory effect on tea blackspot pathogenic fungi of tea by nano-Mg(OH)₂ was studied. The synthesized nano-Mg(OH)₂ was characterized by X-ray powder diffraction (XRD), and the size of nano-Mg(OH)₂ at (101) direction was 14.5 nm according to the Scherrer equation. Scanning electron microscopy (SEM) was applied to observe the morphology of nano-Mg(OH)₂, and it was found that the nanoparticles were regular sheets of structure. Based on the morphological characteristics, internal transcribed spacer (ITS) sequence analysis of ribosomal DNA, ITS-specific primers PCR detection and phylogenetic comparison, it was confirmed that the isolated pathogen was Guignardia mangiferae, which was rarely reported in tea. The water and alkaline condition were set as blank and positive control groups, nano-Mg(OH)₂ with different concentrations were used as experimental groups. After plate coating, the growth diameter of fungi was measured by cross-over method and the inhibition rate was calculated after a period of culture. The results showed that nano-Mg(OH)₂ could effectively inhibit the growth of tea fungi, the inhibition rate of 5 mg/mL nano-Mg(OH)₂ on mycelial growth was 48.78% in 3 days, and the inhibition rate of 50 mg/mL nano-Mg(OH)₂ was 100%, which implying that the inhibition rate was increased with the concentration, and the half maximal effective concentration (EC₅₀) of nano-Mg(OH)₂ was 7.63 mg/mL. This study provides scientific basis and technical support for the subsequent research and development of safe and effective nano-preparations.

Ustilago esculenta, a basidiomycete fungus, can infect Zizania latifolia and induce the swelling of tissues near the base of the host plant. The formed edible gall called 'Jiaobai', that is the second largest aquatic vegetable in China. Chitin, the β-1,4-linked linear homopolymer of N-acetylglucosamine (GlcNAc), is the major component of the fungal cell wall and is crucial for the morphogenesis and survival of fungi. The synthesis of chitin is highly conserved and involves several enzymes, of which the chitin synthase (CS) is the key enzyme. Previous study revealed that there are at least 2 but maybe up to 20 chitin synthase genes per species, which were classified into 7 classes. They are basically involved in the whole process of fungal growth and pathogenicity. ClassⅢchitin synthases are crucial for hyphal growth and pathogenicity in some filamentous fungi. In this study, ClassⅢchitin synthases UeCS3.1 (GenBank No. KU302676) in U. esculenta was cloned based on the whole genome sequence of U. esculenta. The genomic DNA of UeCS3.1 was 3 054 bp with one intron. The full length of its ORF was 2 751 bp encoding a protein with 916 amino acids. In addition, it was predicted that UeCS3.1 had 2 catalytic domains of Chitin-synth-1(pfam01644) and Chitin-synth-1N (pfam08407), seven deduced transmembrane regions, with the theoretical isoelectric point of 8.78 and the molecular weight of 103.73 kD by the biological information analysis of protein structure. The phylogenetic analysis showed that the UeCS3.1 has the highest homology with Umchs1 of Ustilago maydis. Then qRT-PCR was used to detect the relative expression of UeCS3.1 during haploid growth and mating process. The expression analysis showed that the expression of UeCS3.1 was up-regulated during haploid growth and mating process and showed a different relative expression level in the T and MT strains. The relative expression of UeCS3.1 in MT strains was higher than that in T strains at 24~48 h during haploid growth process. Furthermore, during the mating process, the relative expression of UeCS3.1 in T strains was significantly higher than that in MT strains at 24 h and it was continuously up-regulated during 0~72 h while that in MT strains tended to be stable at 72 h. So it was speculated that UeCS3.1 was involved in the process of haploid and mating. For further verification, UeCS3.1 deletion strains were constructed by homologous recombination and PEG mediated protoplast transformation. The morphology of UeCS3.1 deletion strains was observed by microscope and growth curve of UeCS3.1 deletion strains was drawn by the values of OD₆₀₀ at 12, 24, 36, 48, 60 and 72 h. The results showed that the morphology and growth curve of UeCS3.1 deletion strains did not change significantly during the haploid phase. During the mating process, the conjugation tubes of UeCS3.1 deletion strains were formed at 36 h while wild type strains were formed at 24 h. Meanwhile, the hypha of UeCS3.1 deletion strains was shorter than that of wild type strains. Thus, the ability of fusion and hyphal growth of UeCS3.1 deletion strains decreased significantly during the mating process. Totally, these results suggest that UeCS3.1 might be involved in the mating process especially for the hyphal growth in U. esculenta. Above all, this study preliminarily explored the function of the classⅢchitin synthases of U. esculenta, and discussed its role in the mating process, which provided basic materials for the pathogenic mechanism of U. esculenta.

Peste des petits ruminants (PPR) is an acute infectious disease caused by Peste des petits ruminants virus (PPRV), which seriously endangers the healthy development of the world's sheep industry. Since 2013, PPR has been epidemic in many places in China, and the sheep industry suffered huge losses. In this study, the specific primers were designed according to the complete sequence of N gene of PPRV Nigeria 75/1 strain (GenBank No. HQ197753) in GenBank, and the full-length sequence of N gene of PPRV Wuwei strain (GenBank No. KY429031) was amplified. Based on sequencing and sequence analysis, the prokaryotic expression vector pET-PPRV-N was constructed and then was transformed into Escherichia coli Rosetta (DE3). Subsequently, the recombinant protein His-PPRV-N was purified and the Balb/C mice (Mus musculus) were immunized. Then the anti-serum against N protein of PPRV was prepared. Finally, the immunogenicity of recombinant protein was analysed by ELISA , Western blot and IFA (immunological fluorescence assay). The results showed that the N gene of PPRV Wuwei strain shared 99.0% homology with 24 PPRV strains isolated from China as well as Mongolia strain 2016. The phylogenetic tree analysis identified that the PPRV Wuwei strain belonged to lineage Ⅳ. The result of SDS-PAGE showed that the recombinant protein His-PPRV-N was successfully expressed in E. coli Rosetta (DE3) with the form of soluble protein, and the relative molecular weight was approximately 57.8 kD. The results of ELISA ,Western blot and IFA indicated that the His-PPRV-N possessed excellent immunogenicity. The results provide basic information for the establishment of the detection method of PPRV and for further study on biological functions of PPRV N protein.

In the nucleus of mammals, chromatins are folded into various three-dimensional structural units, such as chromosome territories, chromatin compartments, topologically associating domains, and chromatin loops, etc. These structural units play key roles in the regulation of gene expression, cell differentiation and disease development. Previous studies suggested that these structural units represented three dimensional conformations of chromatin at different levels, and distributed hierarchically in mammalian cell nuclei. In recent years, with the rapid development of new technologies of chromatin conformation capture, it has been found that these structural units may not be a simple hierarchical architecture in the nucleus. In this review, we summarize the recently developed technologies of chromatin conformation capture,detailly describe the features and functions of different three-dimensional structural units of chromatin folding.Particularly, we focused on the interrelation among different chromatin structural units. This review provides a valuable reference for subsequent studies.

Jiaobai, the swollen stem of Zizania latifolia, formed by infection with the biotrophic basidiomycete fungus Ustilago esculenta, has become the second largest aquatic vegetables in China. Traditional methods of Jiaobai in planting and breeding not only cost a lot of labor and financial resources but also cause variety degeneration, producing inedible grey Jiaobai and male Jiaobai in the fields, which seriously affect the economic benefits. As a typical dimorphic fungus, closely related to Ustilago maydis, compatible haploid strains of scale pathogenicity were first selected and identified from different strains and combinations of U. esculenta status. Next, leaves, leaf sheaths and tubular roots were inoculated, separately to determine the best inoculation site-leaf sheaths. Then, the inoculation method (stem base injection method, tissue infiltration method with stem base and tissue infiltration method) inoculation serious suspension culture concentration pathogen cell (OD₆₀₀ value of 2.0, 1.5, 1.0, 0.5) and the cultivation environment of seedlings after inoculation were screened. Results showed that the appropriate inoculation method was stem base injection method, the appropriate cell concentration was OD₆₀₀ value of 2.0, and the better cultivation environment was kept in greenhouse or transplanted to field in mid-March or mid-September. An optimized artificial inoculation system was established, which could induce the swollen of stems. Firstly, tubular roots of Z. latifolia were germinated in greenhouse under a 12/12 h light/dark cycle at 25/22 ℃ for 10~20 d; Secondly, compatible strains UET1 and UET2 of U. esculenta with an OD₆₀₀ value of 2.0 were syringe-inoculated into the base of seedling stems, kept in greenhouse or transplanted to field in mid-March or mid-September. Under this artificial inoculation system, plant survival rate was about 80% with swollen stem accounts for more than 80%, tissue slice of leaf sheath and stem apex, collected at early infection stages, analyzed by laser scanning confocal microscopy after staining with wheat germ agglutinin-Alexa Fluor 488 and propidium iodide-Alexa Fluor 561, showed that a large amount of invasion hyphae were formed and expanded to stem apex and full of black masses of teliospores. In order to stop the telispores formation, MT strains from white Jiaobai (swollen stem without telispores) and T strains from grey Jiaobai (swollen stem with full of telispores) were collected and analysed by transcriptome. Differentially expressed genes were screened and selected for functional verification. Fortunately, a key gene Itd1 (interfered factors in teliospores development)(GenBank No. MK164419) related to teliospores formation in U. esculenta was found, encoding 852 amino acids and acted only late stages during infection. Itd1 deletion mutants from the compatible U. esculenta strains UET1ΔItd1 (CGMCC No.16723) and UET2ΔItd1 (CGMCC No.16724) showed a defect in telisorores formation for inoculation. Combining artificial inoculation method and successful genetic modification, an artificial system for U. esculenta-induced swollen stem formation without black teliospores was established. The above study provided a reliable theoretical and technical basis to establish a new efficient and stable breeding way for white Jiaobai. At the same time, it laid a foundation for further research on the dimorphism transition, filamentous growth and teliospores formation of U. esculenta.

Lepidopteran pests cause huge losses to agricultural production every year, and long-term application of chemical pesticides cause serious damage to the ecological environment. Bacillus thuringiensis(Bt) is widely used as microbial insecticide because of its high insecticidal activity and environmental friendliness. The aim of this study was to isolate Bt strains from the soil of Liaoning province and to characterize them on the basis of the morphology of insecticidal crystal proteins (ICPs), the cry gene, the ICPs profiles and the insecticidal activities against lepidopteran pests. The temperature screening method was applied to isolate the wild Bt strains. The cry genotypes were analyzed by restriction fragment length polymorphism PCR (PCR-RFLP). The ICPs profiles were analyzed by SDS-PAGE, and the isolates were tested for their insecticidal activities against 5 species of Lepidopteran pests. A total of 524 soil samples were collected from different areas of Liaoning province, and 66 wild Bt strains were isolated, with a prevalence rate of 12.6%. The ICPs of the strains exhibited varied shapes such as rhombus, sphere, short biconical and irregular shape. The isolated strains expressed ICPs, including 130, 90 and 60 kD proteins. The insecticidal toxicity test showed that Bt strains DD229, DD214-2, YK524 and YK540 had high activities against the tested Lepidoptera pests. The results showed that Bt was abundant in the soils of Liaoning province. And 4 strains that showed high insecticidal activities against Lepidopteran pests which would have good research and application value.