杂色蚕豆新品种“清蚕2号”选育初探

2010年夏在慈溪市白沙路鲜食白花大粒蚕豆大田中发现因小黑蚕豆和白花大粒蚕豆自然杂交而产生变异的F0代变异单株，然后经F1到F5代的连续单株选择，经过F6代的株系选择和F7代的优系繁育，育成了杂色蚕豆新品种——清蚕2号。其株高约90 cm，分枝6～7个，花瓣上部白色，花托处红色，每荚1～3粒， 3粒荚占比仅10%，干豆形状短圆，干豆色泽底色为淡黄色，其侧面有八字形棕褐色条带，豆眼黑色，百粒质量为100 g。“清蚕2号”株型紧凑，植株中等，整齐度高，田间性状稳定，长势较旺盛，抗病性较强，适应性、抗逆性强，结荚多，产量高，品质好。     

Heat,wheat and CO2: The relevance of timing and the mode of temperature stress on biomass and yield

Atmospheric CO₂ enrichment affects C3 crops both directly via increased carbon gain and improved water use efficiency and indirectly via higher temperatures and more frequent climatic extremes. Here we investigated the response of spring wheat (Triticum aestivum L. cv. Triso) to CO₂ enrichment (550 vs. 380 µmol/mol) and heat, applied as a constant +4°C increase or a typical heat wave either before or after anthesis, or as two typical heat waves before and after anthesis. We applied a climate chamber approach closely mimicking ambient conditions. CO₂ enrichment increased above‐ground biomass and yield by c. 7 and 10%, but was not able to compensate for adverse heat stress effects, neither before nor after anthesis, with few exceptions only. Yield depression due to heat stress was most severe when two heat waves were applied (?19%). This adverse effect was, however, compensated by CO₂ enrichment. Applying heat stress before or after anthesis did not exert different effects on yield for both +4°C warming and heat wave application. However, +4°C depressed yield more than a heat wave at ambient CO₂, but not so at elevated CO₂. Thus, the interactive effects were complex and prediction of future wheat yield under CO₂ enrichment and climate extremes deserves more attention.     

Effects of antimicrobial peptides on growth,morphology of foregut villi and related genes mRNA expression in the common carp (Cyprinus carpio)

A 60‐day feeding trial was conducted to examine the effects of different levels (0, 100, 200, 400 and 600 mg/kg) of antimicrobial peptides on growth, protease activity of foregut, the morphology of foregut villi and related genes mRNA expression level in the common carp (Cyprinus carpio). The results showed that the feed of antimicrobial peptides promote common carp growth, and the optimal dosage of antimicrobial peptides is 200–333 mg/kg in the common carp feed. The protease activity of 200 and 400 mg/kg groups were significantly higher than the control and other groups (p < 0.05). The foregut villus height with 100, 200 and 400 mg/kg antimicrobial peptide groups were significantly higher than control group (p < 0.05). The crypt depth of 200 and 400 mg/kg antimicrobial peptide groups were significantly lower than control group (p < 0.05). The ratio of villus height and crypt depth of 100, 200 and 400 mg/kg antimicrobial peptide groups were significantly higher than control group (p < 0.05). The ratio with 600 mg/kg group was significantly lower than the control group (p < 0.05). The IGF‐I gene expression level of 200 mg/kg and 400 mg/kg groups were significantly higher than the control group and 600 mg/kg group (p < 0.05). The IL‐1β gene expression level of 100 mg/kg and 200 mg/kg groups were significantly higher than the control group (p < 0.05). These results indicated up‐regulation of growth and immune related genes in antimicrobial peptides fed common carp. Correlation analysis showed that IGF‐I mRNA and IL‐1β mRNA were positively correlated with SGR. IL‐1β mRNA and FCR were significantly negative correlated. It indicated that growth and immune gene common regulated the growth of the carp under antimicrobial peptides intervention. In conclusion, antimicrobial peptides can improve growth and related genes mRNA expression in the common carp. Further studies using molecular biological technique or immunologic methods are required to conclude that antimicrobial peptides are beneficial in common carp.     

Selection of a new scallop strain,QN‐2, from the backcross of Peruvian scallop × bay scallop F1 hybrids with bay scallops

Cross‐breeding has been successfully applied in agriculture and aquaculture industries to improve important production traits. In our previous studies, we produced the F₁ hybrids with high heterosis in growth by crossing the bay scallops with the Peruvian scallops and selected a new strain, Bohai Red, from the F₁ hybrids. In this study, we selected another new strain from the backcross (BC₁) of the F₁ hybrids with bay scallops. The base cohort (G₀) was size‐selected from the BC₁, and the G₁, G₂, G₃ and G₄ cohorts were produced with size‐selected brood stocks from the G₀, G₁, G₂ and G₃ cohorts respectively. Our results showed that average shell height, shell length, shell width and whole body weight of the G₄ cohort were increased by 16.8%, 16.8%, 11.5% and 43.6%, respectively, compared with the unselected control group of bay scallop. This new strain has now become one of the major cultured Argopecten scallops in northern China waters and was later named QN‐2.     

Orf virus circulation in cattle in Turkey

Orf virus (ORFV) causes contagious skin disease that mainly affects sheep and goats with zoonotic potential. However, there is not enough information about the association between ORFV and occurrence of skin disease in cattle. The present study describes outbreaks of ORFV infection in cattle in different provinces that are located in the Aegean, Central Anatolian and Mediterranean regions of Turkey. During the months of June and August 2017, vesicular fluid and scab samples were collected from cattle which had proliferative skin lesions. First, presence of lumpy skin disease virus (LSDV) and bovine herpesvirus 2 (BoHV-2, known as the causative agent of pseudo-lumpy skin disease) were investigated by real time PCR and PCR, respectively. Then, samples tested for the presence of parapoxviruses by PCR using primers specific to major envelope protein gene (B2L). Parapoxvirus DNA was detected in investigated samples whereas LSDV and BoHV-2 DNA were not detected. The analysis of the B2L gene sequences revealed that cattle were infected with ORFV. The isolates in the present study shared 100% sequence identity at the nucleotide and amino acid level when compared with previously characterised Turkish field ORFV isolates from goats in 2016. Results of the study show unusual infection of cattle with ORFV, and suggest that ORFV jumps the host species barrier from goats to cattle.     

H3N2犬流感病毒M1蛋白的原核表达及鉴定

为制备犬流感病毒(H3N2) M1蛋白纯品,针对M1基因序列设计引物,用聚合酶链式反应(PCR)扩增目的基因片段,扩增产物克隆至表达载体pET-SUMO中并转化至宿主菌BL21(DE3),诱导表达目的蛋白,探索纯化工艺,制备目的蛋白,并用Western blot检测纯化的M1目的蛋白。通过PCR成功扩增出大小为771 bp的M1基因,成功构建p ET-SUMO-M1表达载体,表达的融合蛋白相对分子量为41 kD,主要以可溶形式表达,纯化后获得蛋白纯品,Western blot检测显示用M1蛋白(28 k D)免疫小鼠制备的多抗能与制备的蛋白纯品发生特异性反应,从而证明蛋白纯品为M1目的蛋白。试验制备出的M1蛋白纯品可为进一步制备通用型抗犬流感病毒抗体提供纯品抗原。     

传染性法氏囊病病毒主要结构蛋白VP2、VP1和VP2-VP1串联基因的原核表达及其初步应用

为获得传染性法氏囊病病毒(IBDV)特异性抗体检测用抗原VP2、VP1及VP2-VP1蛋白,分别设计引物扩增IBDV野毒株NN1172的VP2和VP1基因,并扩增VP2和VP1基因中抗原性和亲水性较好的重要区域,通过PCR扩增基因串联方法对截短的VP2和截短的VP1基因进行串联,首次获得VP2-VP1串联基因,并对VP2、VP1和VP2-VP1串联基因进行了原核表达和鉴定。结果成功构建了原核表达载体pET-VP2、pET-VP1和pET-VP2-VP1;诱导表达条件显示,3个重组质粒分别转入BL21菌株后经0.05 mmol/L IPTG诱导表达,分别得到分子量为69、114和63 kDa的VP2、VP1和VP2-VP1重组蛋白,且均以包涵体形式表达,3个重组蛋白分别于诱导后5、3和6 h时表达量最多。Western blot结果显示,表达的VP2、VP1和VP2-VP1蛋白与鸡抗IBDV阳性血清均具有良好的反应原性。以纯化的VP2、VP1和VP2-VP1蛋白作为包被抗原对传染性支气管炎病毒(IBV)、呼肠孤病毒(ReoV)、禽白血病病毒(ALV)和新城疫病毒(NDV)4种阳性血清检测均为阴性,表明所获得的纯化蛋白具有高度的特异性;对免疫了IBD灭活疫苗,IBD基因工程疫苗和IBD弱毒疫苗的商业鸡群进行抗体检测,结果均能显示疫苗免疫后机体抗体水平的变化趋势。本研究表明利用该原核表达系统所表达的3个蛋白均具有良好的免疫反应活性,为IBDV特异性抗体的检测和新型亚单位疫苗的研发奠定基础。     

“金花散茶”及“金花菌粉”对被动吸烟小鼠肺组织JAK2/STAT3炎性及磷酸化蛋白表达的影响

为探究“金花散茶”及其“金花菌粉”对被动吸烟(Cigarette smoking environment,CSE)小鼠肺组织受损的预防及修复机制,建立C57BL/6小鼠CSE模型,以600 mg∙kg^-1剂量的金花散茶茶汤(Eurotium cristatum tea extract,ECTE)及金花菌粉浸提液(Eurotium cristatum powder extract,ECPE)进行灌喂处理。与CSE模型组相比,小鼠灌喂ECPE和ECTE后,肺组织病理学切片显示其可保护小鼠肺组织形态结构完整;酶联免疫分析显示,灌喂ECPE和ECTE可显著抑制小鼠血清IL-6、IL-8、IL-1β、IFN-γ和TNF-α表达量上调;Western blot结果表明,灌喂ECPE和ECTE对小鼠肺组织p-JAK2、p-STAT3、p-JAK2/JAK2、p-STAT3/STAT3高表达起到抑制作用。以上研究结果表明,灌喂ECPE、ECTE对CSE肺受损小鼠具有明显保护作用,总体趋势为ECPE组优于ECTE组、预防组优于治疗组。     

伐地那非增加EGCG-β-乳球蛋白纳米粒对肝癌细胞的抑制作用

较高浓度的EGCG才能抑制癌细胞的增殖,通过纳米化和EGCG与其他药物的联合使用是提高EGCG生物活性的重要策略。本研究将EGCG和伐地那非（VD）同时包埋于β-乳球蛋白（β-Lg）纳米载体中,制备出EGCG-VD-β-Lg纳米粒（EVβ-NPs）,体外试验证实,EVβ-NPs能提高人肝癌细胞（HepG2细胞）中Caspase-3活性,使HepG2细胞在S期产生明显的阻滞,诱发细胞核分裂,从而导致HepG2细胞凋亡。研究结果表明,将EGCG与微量的VD联合使用,并通过纳米化包埋可以显著提高EGCG的抗癌活性。这一方法在EGCG抗癌制品的开发方面具有潜在的价值。     

H9N2禽流感病毒全基因组密码子使用偏好性及影响因素分析

【目的】探究H9N2禽流感病毒(AIV)全基因组的密码子使用偏好性及影响因素。【方法】选取2010—2018年国内H9N2 AIV流行毒株的全基因组为研究对象,分析其碱基组成特性、最优密码子、密码子使用偏好性的影响因素以及病毒对宿主密码子使用模式的适应性。【结果】H9N2 AIV的全基因组中AU含量高于GC。大部分最优密码子以A或U结尾,有效密码子数(ENC)平均值为52.86,提示存在密码子使用偏好性但偏好性较低。密码子使用偏好性主要受到突变压力和自然选择的共同作用,其中自然选择(所占比例为61.79%～76.15%)作用大于突变压力(所占比例为23.85%～38.21%)。H9N2 AIV对人Homo sapiens的密码子适应指数平均值为0.739～0.741,提示H9N2AIV禽流感病毒可能已适应人类的密码子使用模式。【结论】本研究为H9N2 AIV的基因进化分析、已有疫苗的密码子优化和新型疫苗(密码子去优化疫苗)研制提供了理论依据。