The occurrence of cow (Bos taurus) mastitis has restricted the development of dairy industry. It damages the cow health and reduces the quality of dairy products.The main cause of mastitis is pathogenic microorganism. Pathogenic microorganisms could induce cell apoptosis and thus interfere the development of the disease process. The purpose of this study was to explore the expression of cytokines and receptors, and the level of cell apoptosis in bovine mammary epithelial cells (BMECs) infected by Staphylococcus aureus and Escherichia coli. This study was intended to explore the S. aureus and E. coli (with fabrication: Oli) infected BMECs on the expression of cytokine. The BMECs were cultured in vitro and identified by immunohistochemistry, then the purified fifth generation of BMECs were selected and infected S. aureus or E. coli with the concentration of 0, 101, 102, 103, 104 CFU/mL, respectively. By the formula computing, the best infected quantity of S. aureus and E.coil was 0.6 mL; and the number of cell apoptosis was detected by one-step TUNEL (terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling). The expression of interleukin-1β (IL-1β), IL1RⅠ and TNF-α were detected by means of qRT-PCR detection. The results showed that cells expressed not only keratin 18 also beta casein. In addition, the BMECs didn't express vimentin. The cultured cells were identified as BMECs. The highest mRNA expression of IL1RⅠ, IL-1β and TNF-α were at the S.aureus concentrations of 102 CFU/mL, 104 CFU/mL and 103 CFU/mL, respectively. The highest mRNA expression of IL1RⅠ, IL-1β and TNF-α were at the E. coli concentrations of 101 CFU/mL, 103 CFU/mL and 104 CFU/mL, respectively. The apoptosis rate was the highest at 104 CFU/mL with a concentration of cells treated by S.aureus or E. coli. The result indicated that different concentrations of bacterial infecting BMEC could increase the expression of cell factor, while decreased the expression of the receptor, which led to apoptosis of BMEC. This consequence could provide scientific basis for the pathogenesis of mastitis caused by bacterial infection .

Bovine (Bos taurus) herpes virus type I (BHV-1) could cause infectious bovine rhinotracheitis (IBR) and result in economic losing in cattle raising industry. Extensive application of gene-deleted vaccine could significantly decrease BHV-1 infection. In order to construct gene deleted vaccines more efficiently, the present study combined CRISPR/Cas9 gene editing and homologous recombination technique to exchange the glycoprotein E (gE) gene of BHV-1 with green fluorescent protein gene (EGFP), named as BHV-1 gE-/EGFP+. Subsequently, the EGFP gene in BHV-1 gE-/EGFP+ was directly removed by specific small guide RNA (sgRNA), and the gE deleted virus was named as BHV-1 ΔgE. Notably, in vitro biological characteristics analysis showed that the BHV-1 ΔgE had similar growth characteristics and plaque morphology when compared to the parental BHV-1. In vivo animal immunization experiments showed that the CRISPR/Cas9 modifed gE deletion virus did not change the immunogenicity of BHV-1. Thus BHV-1 ΔgE could be used as a candidate for novel recombinant BHV-1 vaccine. The present study first explored the BHV-1 virus genome editing with CRISPR/Cas9 system which could provide an efficient method for constructing BHV-1 gene deleted strains and developing IBR gene deleted vaccine.

The specific detection method based on the specificity of the connection area between external T-DNA and plant genome has very high specificity and accuracy, which is an important technical means to effectively supervise and manage the transgenic event and its derivatives, and ensure the healthy development of the transgenic industry. In the previous study, 2 transgenic soybean (Glycine max) events were obtained with >78% oleic acid content. To isolate the flanking sequences of T-DNAs with endogenous genes and promote the safety assessment and application of these 2 transgenic events, genome re-sequencing combined with PCR was carried out to analyze the flanking sequences of T-DNAs of these 2 transgenic events. There were 1 and 2 copies T-DNAs in these 2 transgenic events by Southern blot analysis, respectively. Using genome re-sequencing compared to reference genome information based on BWA-Burrows-Wheeler Alignment tool method (http://bio-bwa.sourceforge.net/), flanking sequences of T-DNAs in the transgenic soybean 'E2D9050' and 'EB8072' genomes were isolated. The T-DNA of 'E2D9050' was integrated into the position 51410941 on Chr04 with one copy. The T-DNA of 'EB8072' was integrated into soybean genome with 2 copies in position 38147218 on Chr19 and the pattern of integration was inverted insertion with right borders of these copies interfaced. These results were in accord with the results of Southern blot analysis. According to the flanking sequences of these positions, primers were designed and PCR was amplified to verify these flanking sequences of insertion sites. Event-specific primers were designed based on these fragments and the sensitivities of these pairs of primer were both 0.1%. It provides an accurate and fast method for identification of these kinds of transgenic event.

Mining the quantitative trait loci (QTL) related to the maize (Zea mays) grain quality traits can provide the theoretical basis for molecular breeding. In this study, the population containing 150 recombinant inbred lines ( RILs) derived from Xu178×K12 were evaluated for the content of the protein, starch and oil for 3 years under 7 different environments. The best linear unbiased prediction method (BULP) was used to estimate the BLUP value of each trait with the phenotypic value. Composite interval mapping method (CIM) of WinQTLcart 2.5 were using to scanning the QTLs carried out with logarithm viscosity odds (LOD) is 2.5 for 3 quality traits. In total of 20 QTLs were identified distributed chromosome 1, 2, 4, 5, 6 and 10 according to the QTLs analysis. Nine QTLs for protein were identified, six QTLs for starch were identified and 5 QTLs for oil were identified. The QTLs explained 2.3%~15.7% of phenotypic variation. And the LOD values ranged from 2.52 to 6.50. The QTL at Bin4.07/4.08 was detected under 4 environments and BLUP value. And QTL in Bin6.05/6.06 was detected under 4 environments. As well as 2 QTLs located in Bin5.07 were detected in 3 environments. In this study, the QTL mapped in the marker intervals umc1194~umc2384 on chromosome could detected in multiple environments. The QTL explained 10.3%~15.7% of phenotypic variation. This stable QTL was considered to be major QTL for maize protein content, which was expected that this QTL could be applied on the genetic improvement of maize grain quality. The experiment of QTL analysis associated quality traits of maize grain provides basic data for molecular breeding of maize.

Sucrose is one of the main assimilates in plant photosynthesis process. The effective use of sucrose in plant reproduction plays important roles in the process of fertilization and fruit-setting. Invertase (INV), as a key enzyme of sucrose metabolism, regulate the hydrolysis of sucrose into glucose and fructose. According to the cellular localization, INVs are further divided as cytoplasmic invertase (CIN), vacuolar invertase (VIN), and cell wall invertase (CWIN). In order to understand the physiological and molecular mechanism of sucrose metabolism involved in the process of male sterility of pepper (Capsicum annuum) induced by pollen abortion, the INV enzyme activities, INV genes expression pattern and their correlation with pepper male sterility were analyzed. A pair of pepper male sterile line 131A and maintainer line 131B (fertile) were used as investigating materials in this study. The parameters including INV enzyme activities, VIN and CWIN gene identification in pepper genome, and CWIN encoding gene expression pattern by qRT-PCR, were comparatively analyzed in different tissues and developmental stages of pepper flower. The results showed that the CWIN activities in the big bud and flower of the maintainer line 131B were significantly higher than those in the male sterile line 131A (P＜0.001 and P＜0.01, respectively), which were closely correlated with the high expression level of pepper CaCWIN encoding genes CaCWIN3 and CaCWIN4. According to the results, CaCWIN3 and CaCWIN4 were likely the key functional genes involved in the process of male sterility of pepper. These results shed light on the further investigation of molecular regulation mechanism of capsicum male sterility.

Plant cellulose synthase is one of important glycosyltransferases, which catalytic synthesis a paracrystalline form of H-bonded-β-1, 4-Glc chains. In this study, a unigene that was highly homologous to the Zea Mays cellulose synthase 3 (Ces3) sequence was isolated from the sugarcane (Saccharum officinarum) full-length cDNA library in response to water stress. The full-length sequence was to obtain by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE), contained most 5'-end excluding 80 bp and the whole 3'-end with poly (A), named as ScCes3 (GenBank No. MG324347). ScCes3 had a full-length cDNA sequence of 3 625 bp and contained a contained open reading frame (3 225 bp), encoding 1 074 amino acids. Bioinformatics prediction indicated that ScCes3 was stable amphoterin protein located in cell plasmalemma, and had no signal peptide but with random coil according to its secondary structure. The ScCes3 protein contained the conservative domains of plant cellulose synthase proteins. qRT-PCR results indicated the expression of ScCes3 could be found in leaf, leaf sheath and stem. The high expression level in stem, especially with the highest expression level in No.5 stem, and lowly in leaf. This study has established a foundation for future research on gene expression and funtion analysis.

Abstract Respond to low-phosphorus stress, the plasma membrane H+-ATPase plays an important role in plant. In this study, a PM H+-ATPase gene sequence was screened from the transcriptome of Chinese fir . The sequence of PM H+-ATPase were cloned from three Chinese fir clones(X6,3 and 39) respectively. The resulting sequence was 2898bp in length, containing a 2862bp complete open reading frame encoded 953 amino acids residues, named as ClHA2 (Accession No.). This amino acids sequence analysis showed that ClHA2 was highly conserved with PM H+-ATPase of other plants. The predicted relative molecular weight of the protein was about 105.05 kDa, the theoretical isoelectric point (PI) was 6.38.The protein contains ten transmembrane domains,and the secondary structure of ClHA2 was mainly consisted of α-helix. The results of quantitative real-time PCR showed that ClHA2 gene of three Chinese fir clones was expressed in roots ,stems and leaves under low-phosphorus stress,and its expression was inhibited to some extent.

The fibroblast growth factor 10 gene (FGF10) is a member of the fibroblast growth factor family, it plays an important role in the development and metabolism of white adipose tissue. In order to further reveal the role of FGF10 gene in the regulation of intramuscular fat in Congjiang Xiang pigs (Sus scrofa), in this experiment, four pairs of siRNA interference sequences and a pair of negative control sequences were designed by online software,and were connected to pGPU6/GFP/Neo vector. The successfully constructed interference vectors were transfected into Congjiang Xiang pig intramuscular preadipocytes and the best interference vector was screened by qRT-PCR. The expression of FGF10 gene was inhibited by the best interference vector. And then, the expressions of peroxisome proliferator activated receptorγ (PPARγ), adipose triglyceride lipase (ATGL),adiponectin (ADIPOQ), fatty acid binding protein 4 (FABP4), fatty acid synthase (FAS), lipoprotein lipase(LPL), acetyl-CoA carboxylase (ACC) and Hormone-sensitive lipase (HSL) gene were detected by qRT-PCR.. The results showed that the interference vector of the FGF10 gene was constructed successfully, and the optimal interference vector was screened with the interference efficiency of 74.30%. After inhibiting the expression of FGF10 gene, the expression of ACC, ATGL, ADIPOQ, FABP4, FAS, LPL, PPARγ and HSL were decreased. The expression of ACC and PPARγ genes were significantly lower than the control group (P＜0.05), and the expression of ATGL, ADIPOQ, FABP4, FAS and LPL genes were significantly lower than control group (P＜0.01). There was no significant difference between the relative expression of HSL gene and the control group (P＞0.05). In this experiment, the constructed interference vector of FGF10 gene was successfully transfected into the intramuscular preadipocyte cells of Congjiang Xiang pig, and the procine intramuscular preadipocytes differentiation can be inhibited by sliencing of FGF10 gene in vitro, which might be related to down-regulation of fat differentiation genes. This study provides foundation for further research on the regulation mechanism of FGF10 gene on fatty deposits.

Delay in maternal transcript degradation of DNMT1 (DNA (cytosine-5)-methyltransferase 1cytosine-5)-methyltransferase 1) in porcine (Sus scrofa) somatic cell nuclear transfer embryos compared with in vitro fertilized counterparts was confirmed, thereafter anti-DNMT1 siRNA was injected into embryos at one-cell stage to evaluate the effect of DNMT1 knockdown on the developmental rate of embryos and the mRNA levels and DNA methylation levels of several developmentally important genes. Though a higher proportion of blastocysts was obtained in 50 μmol/L siRNA injected embryos (30.43%) compared with mock injected embryos (23.01%) and untreated controls (25.74%), while the difference was not statistically significant (P= 0.19). In the case of genes expression, inhibition of DNMT1 significantly increased the mRNA levels of POU5F1 at four-cell stage and of NANOG at the blastocyst stage (P＜0.05). In the aspect of DNA methylation, inhibition of DNMT1 reduced the DNA methylation level of POU5F1 at four-cell stage and decreased the general methylation to the level of in vivo produced embryos at the blastocyst stage, whereas over-depressed DNMT1 disrupted the imprinted methylation of a long noncoding RNA H19. This study suggests that mild inhibition of over-expressed DNMT1 facilitates pluripotent genes transcription and demethylation reprogramming of porcine cloned embryos, furthermore provides a scientific reference for further research on function mechanisms of DNMT1 during early embryogenesis.

Long noncoding RNA (LncRNA) is involved in a variety of biological processes in mammals, and the current research on its role in hair follicle formation process is relatively small, therefore, studying the effect of LncRNA on hair follicle development has become a new direction to understand the biological process of hair follicle formation. In order to verify the RNA-Seq results of skin tissue in the non-period of the development of the Subo Merino sheep (Ovis aries) and the difference in the expression of predicted target genes, the expression of two LncRNA 00202353 and LncRNA 00316173 and their target genes in skin tissue, which were differentially expressed in skin tissue during the development of Subo Merino sheep and their target genes associated with hair follicle development were detected by qRT-PCR. The results showed that the expression level of LncRNA 00202353 and its target gene secreted frizzled-related protein 2 (SFRP2) at 85 day of embryonic stage was extremely significantly higher than 65 and 135 d of embryonic stage, 7 and 30 d after birth (P＜0.01). The expression level of LncRNA 00316173 and its target gene insulin-like growth factor 2 (IGF2) at 65 d of embryonic stage was extremely significantly higher than 135 d of embryonic stage, 7 and 30 d after birth (P＜0.01). Analysis of the hair follicle developmental stage of the Subo Merino sheep was divided into developmental and maturity stages, the expression levels of LncRNA 00202353 and its target gene SFRP2, LncRNA 00316173 and its target gene IGF2 were extremely significantly higher in the hair follicle development than in the hair follicle maturation (P＜0.01). It indicatd that LncRNA 00202353 and LncRNA 00316173 and their target genes were involved in the development of hair follicles. These resullts provide important reference value for further studies on the development of hair follicles in LncRNAs, and supply research clues for exploring the biological processes of the periodic changes of ultrafine woolen sacs.

Bacitracin methylene disalicylate (BMD), as the second generation of bacitracin feed additive, can promote the growth and improve the immunity of livestock. This study aims to investigate the dynamic effect of BMD on the immune function of rabbits, 300 New Zealand weanling rabbits (35-day-old) were divided into BMD high (BMD H, 100 mg/kg), BMD middle (BMD M, 50 mg/kg), BMD low dosage group (BMD L, 25 mg/kg), Bacitracin Zinc group (BZ, 50 mg/kg), and blank control group (C, no addition). Studies in this group have confirmed that BMD could promote the growth of rabbits. At the end of the 35-day trial, the contents and mRNA expression of interleukin 2 (IL-2) and immunoglobulin G (IgG) in cecum and duodenum were detected by enzyme linked immunosorbent assay (ELISA) and qRT-PCR assay. The contents of IgG and IL-2 in cecum in BMD and BZ group were significantly higher than C group (P＜0.05). The contents of IL-2 in duodenum in BMD H group were significantly higher than that C group (P＜0.05). The correlation analysis showed that the contents of IL-2 and IgG in the cecum were positively correlated with the gene expression levels (P＜0.05). The results showed that the addition of BMD to diets could increase the content of IL-2 and IgG, and upregulate mRNA expression. The effect of 50~100 mg/kg BMD in dietary supplementation is equivalent to the effect of BZ. This study further confirmed that BMD could enhance the immunity of rabbits, and provide evidence for its application in rabbit industry.

Euryhalinous teleosts can survive in certain range saline-alkali water environment. The Gill is considered to be the main organ which is responsible for ion transport , maintain osmotic pressure balance and acid-base equilibrium.This study was to screen and identify the differential expressed proteins in gill of the Red tilapia (Oreochromis mossambicus ♀× O. niloticus ♂) by protein chips and mass spectrometry techniques, then verified by using immunohistochemistry and Western blot techniques, and to provide protein theoretical basis for alkali-resistant mechanism. There were 229 differential expressed protein spots detected (change multiple≥1.5), which contains 41 up-regulated proteins and 49 down-regulated proteins. Among them, 2 proteins were identified by mass spectrometry from 5 differential expressed proteins: 14-3-3 protein beta/alpha-A-like isoform X1 (14-3-3), Dna J homolog subfamily β member 11-like (Dna J).The immunohistochemistry results showed that 14-3-3 and Dna J were expressed in the gill base of Red tilapia at fresh water and alkalinity water, and showed a trend of first decrease and then the positive reaction increased significantly with increasing alkalinity. The results of Western blot showed that in the fresh water and alkali water, 14-3-3 and Dna J proteins were both expressed in the gill. With increasing alkalinity, the protein expression was the same as the immunohistochemistry. The expression of 14-3-3 protein showed a trend of increase with the increase of alkalinity concentration, and the expression of Dna J protein was significantly increased at alkalinity water which compare with fresh water. In the 4 alkalinity group , 14-3-3 protein reduced in the early stage of stress,then increased, and reached the lowest value at 12 h, and the peak value rised at 72 h. While the expression of Dna J protein was significantly increased at 96 h (P＜0.05). The results suggest that red tilapia is able to cope with protein activitity and stability in the body by activating the protein 14-3-3 and Dna J protein in the gill tissue, thereby maintaining the body protein normal function stability.The expression of 14-3-3 and Dna J proteins in gill of red tilapia was increased under alkalinity stress. Thus it would have great significance to support that 14-3-3 and Dna J proteins are both participated in the response of alkalinity stress.The present study gives more information to learn the mechanism of alkalinity stress response in red tilapia.

Corythucha ciliata is a major and specific pest of Platanus acerifolia. The objective of this study is to establish the antennal transcriptome database of the sycamore lace bug (Corythucha ciliata), and to obtain the olfactory-related genes in C. ciliata. The antennal transcriptome of male and female adults of C. ciliata was sequenced using Illumina HiSeqTM 4000 platform and bioinformatics analysis. A total of 65 943 Unigenes were obtained, with a mean length of 1 036 bp and an N50 of 1 809 bp. Through a similarity search, 16 600 Unigenes were annotated, most Unigenes (23.12%) were annotated to a database of orthologous groups of gene (eggNOG) database. All Unigenes against the NCBI Non-redundant Protein Sequences (Nr) database had the highest similarity (14.40%) with those of Zootermopsis nevadensis. Moreover, according to Gene Ontology (GO) database, 6 435 Unigenes were broadly divided into 52 branches of 3 categories: 16 271 Unigenes to be related to biological processes categories, 10 285 Unigenes related to cellular components, and 7 989 Unigenes related to molecular functions, and more Unigenes were annotated to be related to metabolic process, cellular process, catalytic activity and binding activity. Gene expression analysis showed that a total of 1 116 genes expressed differentially, involving in 1 096 genes up-regulated genes and 20 down-regulated genes. According to clusters of orthologous groups of proteins (KOG) database, 10 335 Unigenes were divided 25 categories, 2 515 and 1 233 Unigenes were related to general function prediction only and signal transduction mechanisms, respectively. By searching Kyoto Encyclopedia of Genes and Genomes (KEGG) database, 6 309 Unigenes were assigned to 255 metabolic pathways. Including 346 differentially expression genes, among these, the most genes (20) were involved in metabolic pathways of carbon. By further screening and identification, 194 olfactory-related genes of C. ciliata were obtained, including 77 odorant receptors (OR) genes, 64 odorant degrading enzymes (ODE) genes, 26 odorant binding proteins (OBP) genes, 11 ionotropic receptors (IR) genes, 14 chemosensory proteins (CSP) genes and 2 sensory neuron membrane proteins (SNMP) genes. A total of 65 943 Unigenes, 5 404 Unigenes with higher expression abundance (FPKM＞10) and 12 726 Unigenes with an FPKM (fragments per kilobase per million reads) value of 0. There were 2 OBP genes among the top 50 genes with higher expression abundance, and the highest expression level of OBP gene with an FPKM value of 953. This study preliminarily clarified the whole expression pattern of the antennal transcriptome of male and female adults of C. ciliata, which will provide a molecular foundation for further studying the mechanism of olfactory regulation of C. ciliata.

Rhynchophorus ferrugineus is a major quarantine pest that damages the coconut trees (Cocos nucifera) and other palm plants (Arecaceae). In this study, the strain Yel-8 was isolated from the corpse of R. ferrugineus, which had insecticidal activity against the larvae of R. ferrugineus which was suffered from sepsis and identified by morphological identification, 16S rDNA, physiological and biochemical tests, and drug susceptibility analysis, respectively. The results showed that Yel-8 was in line with the characteristic of Yersinia entomophaga in L-arabinose, rhamnose, xylose, sucrose, aesculin, and cellobiose metabolism, whereas opposed to Yersinia only in sorbitol metabolism. Therefore, this strain was identified as a new strain of Yersinia genus. The bioassay of Yel-8 against R. ferrugineus was performed using feeding and injection methods, and the median lethal concentration (LC50) of Yel-8 against R. ferrugineus was 3.184×108 and 5.1295×106 CFU/mg, respectively. The chitinase genes Chi1 and Chi2 were cloned from the Ye1-8 strain and the proteins were expressed for bioassay. The results showed that the Chi1 and Chi2 proteins had no insecticidal activity against R. ferrugineus alone, thus suggesting the necessity of complete toxin complex for toxicity. Yel-8 exhibited insecticidal activity may provide an ideal material for the construction of the biophytes against R. ferrugineus.

To explore the effect of gene expression on the fate of root tip cells, different types of root cells need to be grouped for further analysis. In the present study, the protoplasts of young root tips of Zea mays were obtained by enzymatic hydrolysis, and analyzed and sorted by fluorescence staining and flow cytometry. The results showed that the optimum method for obtaining protoplasts of 4~5 d young maize root tip tissue by enzymatic hydrolysis included combination of 1.0% cellulase and 0.3% pectinase, optimum osmotic pressure with 10% mannitol, optimum enzymatic hydrolysis time with 4 h and optimum pH with 5.8. The protoplasts extracted under the above optimum conditions were purified and the impurities such as cell fragments in the protoplast suspension were effectively removed. The purified protoplasts were stained by Propidium iodide (PI) and Fluorescein diacetate (FDA), and the observation under fluorescence microscopy showed that the protoplasts were highly active. For further sorting by flow cytometry, different fluorescent dyes were used. The staining effect of Hoechst33342 on active protoplasts was better than that of 4', 6-diamidyl-2-phenylindole (DAPI), most protoplasts in the system were stained with high resolution. At the same time, for Hoechst33342, the optimum dyeing temperature was 38 ℃, and the optimum dyeing pH was 10. The study provides basic data for further related researches on root tip cells in plants.

Small Meishan pig (Sus scrofa) is a unique local pig breed in the Taihu River Basin which has the characteristics of delicious meat. There is the problem of counterfeiting meat products in the market, while the traditional identification technology has some limitations in practical application. The present study is to develop a new method to distinguish Small Meishan pig from other pig breeds or strains of Taihu Basin and common western pig breeds by using the third generation molecular marker called Single Nucleotide Polymorphism (SNP). The data of genotyping by genome reducing and sequencing (GGRS) of 7 local breeds (strains) in Taihu Basin (Middle Meishan pig, Small Meishan pig, Erhualian pig, Mi pig, Jiaxinghei pig, Fengjing pig and Shawutou pig) and 5 imported pig breeds (Duroc, Landrace, Yorkshire, Piltland, and Baxia) was used. Five SNP sites only in Small Meishan pig were selected and Sanger sequencing was used to verify the results. It was showed that the 5 SNP sites had high mutation rate in Small Meishan pig. By using the combination of 5 candidate sites, the identification probability could be up to 99.35% and the accuracy was 100% with no false positive identification. The present study provides a basic principle for the protection of genetic resources and identification of fake and inferior pork products in Small Meishan pig.

Microbial whole-cell sensor has been widely used to assess bioavailability and risk of toxic elements, but their environmental use is still limited due to the low sensitivity and poor selectivity. Here, Escherichia coli BL21 (DE3) was employed as a bacteria strain of the sensor, the expression vector pETDuet-1 as an initial skeleton, the DNA fragment O/P-cadC consisting of the promoter/operator of the cad operon and cadC gene as a sensing module, and gfp as a reporter gene to construct a new type of microbial whole-cell sensor in response to Cd2+. The GFP fluorescence of the sensor cells induced by Cd2+ was measured by fluorescence emission spectroscopy and fluorescence microscopy, and the sensitivity changes of the biosensor induced by Cd2+ at low temperature was analyzed. Results showed that a GFP-based microbial whole-cell sensor E. coli BL21(DE3)_pO/P-cadC::gfp was successfully developed, which exhibited strong green fluorescence exposed to Cd2+ and dose-dependent effects of green fluorescence on the concentration of Cd2+. The lowest detectable concentration of the biosensor at 37 ℃ was 1 mmol/L, whereas the lowest detectable concentration decreases to 1 μmol/L at 25 ℃, and 10 nmol/L at 15 ℃. The sensitivity of the biosensor induced with lower temperature is improved obviously. This study indicated that the constructed E. coli whole-cell sensor could dose-dependently and selectively respond to Cd2+. The temperature has an important effect on the fluorescence intensity of the sensor cells, and the sensitivity of the sensor can be improved significantly at low temperature. This provides reference for the detection of bio-availability of Cd2+ in the environment by using the microbial whole-cell sensor.

The number of vertebrae is an economically important trait in livestock. Multi-vertebrae means increasing in numbers of animal's thoracicae and lumbales. Animals can improve performance in the size, meat production, gross production, skin area, adaptability and feed utilization with the increase of their vertebral number. Animal breeds with multi-vertebrae can be cultivated by directional selection because of the high heritability of vertebral number trait. Many achievements of studies on vertebral number trait have been made in swine (Sus scrofa), whereas the results were limited in other livestock such as sheep (Ovis aries), and cattle (Bos taurus) and yak (B. grunniens). This paper summarized the advances in gene mapping of vertebral number trait, and briefly described the new opportunities brought by high-throughput sequencing for the study in gene mapping of vertebral number trait in livestock.

Chromosome missegregation in mammalian oocytes and early embryos leads to embryo aneuploidy and early pregnancy loss. In mitotic cells, the microtubules that compose the spindle are mostly nucleated and bipolarized by centrosomes acting as major microtubule organizing centers (MTOCs). However, mammalian oocytes remove their centrosomes for reasons that are still speculative. Spindle assembly needs suppletive pathways (including RanGTP pathway, Augmin pathway and CPC pathway). This paper illuminated the centrosome-independent spindle assembly and related processes in mammalian oocytes, and discussed the differences in spindle assembly and the relationship between spindle assembly and aneuploidy in different mammalian oocytes. This paper provides reference for the mechanism of mammalian aneuploid formation and chromosome aberration.