葡萄是世界上最重要的经济作物之一，改善其主要农艺性状以适应不断变化的农业环境和消费者需求是非常重要的。随着现代生物技术的发展，尤其是分子遗传技术的应用，为选育高产、优质、抗逆和抗病性强的葡萄新品种提供了新方法。目前，通过基因枪和农杆菌介导法已成功将一些葡萄品种进行遗传转化，并通过建立的再生体系获得了转基因葡萄植株。然而，基因型，外植体类型以及培养基的组成成分能够影响葡萄植株再生效率；受体材料、菌株和浓度、筛选标记的选用也会影响其转化效率。本文归纳总结了近年来报道的葡萄植株再生和遗传转化的研究进展，存在的问题和未来的研究方向，以期为葡萄新品种改良提供参考。

重组病毒活载体疫苗是一种能够有效激活特异性和非特异性免疫、可多联多价、安全性好的新型疫苗。动物α疱疹病毒拥有较大的基因组，含有多个不影响病毒复制的非必需区，能够插入接受外源基因并表达相应抗原蛋白；同时具有较广泛的宿主谱，能够在宿主体内复制并持续刺激动物产生对抗相应病原的免疫力，是作为重组病毒活载体疫苗的理想载体。随着基因编辑技术的发展，可通过多种方法构建能够表达外源基因的重组病毒。目前以动物α疱疹病毒为载体的重组病毒活载体疫苗研究已经涉及禽类、猪、牛、羊、伴侣动物等，目前成功构建的多株重组动物α疱疹病毒能免疫后可使动物同时获得对多种疾病的免疫。本文总结了重组动物α疱疹病毒构建方法、外源基因的引入和表达以及动物α疱疹病毒活载体疫苗免疫作用三个方面的内容，包括了最新的基因编辑技术、不同的构建策略及其优缺点、外源基因的选择、插入形式和位点等，并介绍了各种动物α疱疹病毒活载体疫苗的最新研究进展，旨在为新型动物α疱疹病毒活载体疫苗的研究和开发提供一定的参考。

A total of 900 one-d-old Chinese Huainan Partridge Shank chickens were randomly allocated into nine groups with five replicates of 20 each.  Birds were fed with basal diet, basal diet supplemented with 150 mg kg^–1 aureomycin, basal diet supplemented with different proportions of Bacillus licheniformis and Bacillus subtilis, which was 0:1.0×10⁶, 2.5×10⁵:7.5×10⁵, 3.3×10⁵:6.6×10⁵, 5.0×10⁵:5.0×10⁵, 6.6×10⁵:3.3×10⁵, 7.5×10⁵:2.5×10⁵ and 1.0×10⁶:0, respectively.  The duration of the experiment was 56 d.  The results indicated that dietary supplementation of 6.6×10⁵:3.3×10⁵ of B. lichenifornis:B. subtilis improved final body weight, increased the average daily gain, and reduced feed/gain ratio (P<0.05).  The numbers of total Lactobacillus and Bifidobacterium sp. in the caecum significantly increased, and the numbers of Escherichia coli and Salmonella sp. significantly declined compared to that of the control (P<0.05).  Intestinal villous height and villous height to crypt depth ratio of the duodenum, jejunum, and ileum were significantly higher than that of the control, and intestinal crypt depth of the duodenum and ileum was significantly lower (P<0.05).  The total antioxidant capacity, total superoxide dismutase, and glutathione peroxidase ability in plasma significantly improved, while the malondialdehyde concentration in plasma decreased (P<0.05).  Compared to the control, plasma concentrations of ammonia, uric acid and urea nitrogen and the activity of xanthine oxidase were reduced (P<0.05).  In conclusion, an inclusion of 6.6×10⁵:3.3×10⁵ of B. licheniformis: B. subtilis to the diet improved the growth performance, caecal microbiota, plasma biochemical profile, and significantly improved the small intestine morphology, while reducing the mortality rate. 

Many mammalian herpes viruses utilize heparan sulfate (HS) moieties present on cell surface proteoglycans as receptors for cell entry, and this process also requires viral glycoprotein C (gC) homologues.  However, our understanding of the role of gC in facilitating attachment of other alpha-herpes viruses such as the duck plague virus (DPV) remains preliminary.  To study the role of gC during DPV infection, we used a gC-deleted mutant virus (DPV-ΔgC-EGFP).  Examination of the viral copy number by real-time PCR, as well as time course studies of viral adsorption and proliferation revealed that gC was involved in the viral binding to the cell surface.  The affinity of viral glycoproteins (gB-DPV, gC-DPV, and gE-DPV) to HS was assessed using a prokaryotic expression system and HiTrap^TM Heparin HP column chromatography.  In addition, to confirm that gC played a role in the interaction between DPV and HS, viruses were treated with the HS analogue heparin and host cells were treated with its inhibitors heparinase prior to exposure to DPV-ΔgC-EGFP or wild-type strain Chinese virulent duck plague virus (DPV-CHv).  The effects of heparin and heparinase on virus infectivity demonstrated that function of gC on viral adsorption is independent of interactions between gC and heparin sulfate on cell surface.  All in all, this study demonstrated that the gC of DPV can mediate viral adsorption in an HS-independent manner, which distinguish it from the gC of some other alpha-herpes viruses.  Future studies will be required to identify the receptors involved in gC protein binding to cells.  This work provides us a foundation for further studies of examining the roles of gC in the adsorption during duck plague virus infection.

The effects of antibacterial compounds produced by Saccharomyces cerevisiae in Koumiss on pathogenic Escherichia coli O₈ and its cell surface characteristics were investigated.  S. cerevisiae isolated from Koumiss produced antibacterial compounds which were active against pathogenic E. coli O₈ as determined by the Oxford cup method.  The aqueous phases from S. cerevisiae at pH=2.0 (S2) and pH=8.0 (S8) were extracted and tested, respectively.  The organic acids of S2 and S8 were determined by high performance liquid chromatography (HPLC), and the concentrations of killer toxins were determined by enhanced bicinchoninic acid (BCA) Protein Assay Kit.  The minimum inhibition concentration (MIC) and the minimum bactericidal concentration (MBC) of S2 and S8 on E. coli O₈ were determined by the broth microdilution method.  The effects of S2 and S8 on the growth curve of E. coli O₈ were determined by turbidimetry, and the hydrophobicities of E. coli O₈ cell surface were determined using the microbial adhesion to solvents method, the permeation of E. coli O₈ cell membrane were determined by the o-nitrophenyl-β-D-galactoside (ONPG) method.  Aqueous phases at pH 2.0 and 8.0 had larger inhibition zones and then S2 and S8 were obtained by freeze-drying.  The main component in S2 was citric acid and it was propanoic acid in S8.  Other organic acids and killer toxins were also present.  Both the MICs of S2 and S8 on E. coli O₈ were 0.025 g mL^–1, the MBCs were 0.100 and 0.200 g mL^–1, respectively.  The normal growth curve of E. coli O₈ was S-shaped, however, it changed after addition of S2 and S8.  E. coli O₈ was the basic character, and had a relatively hydrophilic surface.  The hydrophobicity of E. coli O₈ cell surface and the permeation of E. coli O₈ cell membrane were increased after adding S2 and S8.  The present study showed that S2 and S8 inhibit the growth of pathogenic E. coli O₈ and influence its cell surface characteristics.

   Recently, a new bacterial top rot disease of maize has frequently appeared in many areas of Yunnan Province, China. The pathogen of the disease was identified as Klebsiella pneumoniae (KpC4), which is well known to cause pulmonary and urinary diseases in humans and animals and occasionally exists as a harmless endophyte in plants. To evaluate the virulence of the maize pathogen to maize and mice, we inoculated maize and mice with routine inoculation and intraperitoneal injection respectively according to Koch’s postulates. The results showed that KpC4 and the clinical strain K. pneumoniae 138 (Kp138) were all highly pathogenic to maize and mice and the strain re-isolated from diseased mice also caused typical top rot symptoms on maize by artificial inoculation. It is highlighting that a seemingly dedicated human/animal pathogen could cause plant disease. This is the first report of K. pneumoniae, an opportunistic pathogen of human/animal, could infect maize and mice. The findings serve as an alert to plant, medical and veterinarian scientists regarding a potentially dangerous bacterial pathogen infecting both plants and animals/humans. The maize plants in the field could serve as a reservoir for K. pneumoniae which might infect animals and probably humans when conditions are favorable. The new findings not only are significant in the developing control strategy for the new disease in Yunnan, but also serve as a starting point for further studies on the mechanism of pathogenesis and epidemiology of K. pneumoniae.

    CD8, a glycoprotein on the surface of T cells, is involved in the defense against viral infection and plays significant roles in antigen presentation and in the antiviral immune response. CD8 is composed of two chains. Of these, the CD8α chain was chosen for the detection because it involved in both the CD8αα homodimer and the CD8αβ heterodimer. Here, we established a double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for specific detection of goose CD8α (goCD8α). The results showed that the optimal coated antibody and antigen dilutions were 1:50 (the antibody titer was 1:12 800) and 1:32 (0.3 ng mL^–1), respectively, while the optimal capture antibody and horseradish peroxidase (HRP)-labelled goat anti-rabbit IgG dilutions were 1:50 (the antibody titer was 1:51 200) and 1:4 000 (the antibody titer was 1:5 000), respectively. The optimal blocking buffer was 5% bovine serum albumin (BSA). The best incubating condition was overnight at 4°C, the best blocking time was 120 min and the best anti-capture antibody working time was 150 min. In addition, the minimum dose detectable by DAS-ELISA was 5×10–3 ng mL^–1. Most importantly, goCD8α expression levels in goose spleen mononuclear cells (MNCs) post-Goose parvoviruse (GPV) infection were found to be significantly up-regulated using the DAS-ELISA method, which was consistent with previous results obtained using real-time quantitative PCR. In conclusion, the DAS-ELISA method reported here is a novel, specific technique for the clinical detection of goCD8α.

Grain size is one of the most important agronomic components of grain yield. Grain length, width and thickness are controlled by multiple quantitative trait loci (QTLs). To understand genetic basis of large grain shape and explore the beneficial alleles for grain size improvement, we perform QTL analysis using an F2 population derived from a cross between the japonica variety Beilu 129 (BL129, wide and thick grain) and the elite indica variety Huazhan (HZ, narrow and long grain). A total number of eight major QTLs are detected on three different chromosomes. QTLs for grain width (qGW), grain thickness (qGT), brown grain width (qBGW), and brown grain thickness (qBGT) explained 77.67, 36.24, 89.63, and 39.41% of total phenotypic variation, respectively. The large grain rice variety BL129 possesses the beneficial alleles of GW2 and qSW5/ GW5, which have been known to control grain width and weight, indicating that the accumulation of the beneficial alleles causes large grain shape in BL129. Further results reveal that the rare gw2 allele from BL129 increases grain width, thickness and weight of the elite indica variety Huazhan, which is used as a parental line in hybrid rice breeding. Thus, our findings will help breeders to carry out molecular design breeding on rice grain size and shape.

Spermatogonial stem cells (SSCs) are unique stem cells in adult body that can transmit genetic information to the next generation. They have self-renewal potential and can continuously support spermatogenesis throughout life of a male animal. However, the SSC population is extremely small, isolation and purification of the SSCs is challenging, especially for livestock animals. It has been confirmed that CDH1 (cadherin-1, also known as E-cadherin) can be expressed in undifferentiated SSCs of mouse and rats, but it has not been verified in sheep. Here, CDH1 was found as a novel surface marker for sheep SSCs. In this paper, sheep anti- CDH1 polyclonal antibodies were prepared and its activity was checked. Using the obtained antibodies and immunohistochemistry analysis, we confirmed that CDH1 can be expressed by SSCs in sheep testis.

The developmental competence of porcine parthenotes cultured in porcine zygote medium-3 (PZM-3) and North Carolina State University-23 (NCSU-23) media was investigated. After in vitro maturation oocytes were electro-activated, and the subsequent developmental competence, rates of apoptotic, fragmented and arrested embryos from the two media were evaluated. Also, the ratio of mRNA expression of Bcl-2 and Bax gene was determined. Results demonstrated that cleavage, blastocyst, hatched blastocyst rates, and blastocyst cell numbers were significantly higher in PZM-3 than in NCSU-23. The rate of fragmented embryos in PZM-3 was lower than in NCSU-23 on d 1 and 3 (P<0.05) while the percentage of arrested embryo was lower in PZM-3 than in NCSU-23 on d 4 and 5 (P<0.05). TUNEL positive signals were higher in NCSU-23 than in PZM-3 from d 3 to 7 (P<0.05). The ratios of Bcl-2 and Bax mRNA expression in embryos were higher on d 5 than on d 3 and 1 in PZM-3 (P<0.05). In contrast, the ratios of Bcl-2 and Bax mRNA expression in embryos on d 1 were higher than on d 3 and 5 in NCSU-23 (P<0.05). Additionally, the ratios of Bcl-2 and Bax mRNA expression in embryos in PZM-3 were higher than in NCSU-23 on d 3 and 5 (P<0.05). In conclusion, lower apoptotic embryo rates and down-regulating Bax together with up-regulating expression of Bcl-2 transcripts may be responsible for the better developmental competence of porcine parthenotes in PZM-3.