Prediction of grain protein in spring malting barley grown in northern Europe

The ability to predict grain protein concentration at harvest (CP) in Swedish malting barley (Hordeum distichum) from observations of soil conditions, sowing day, fertilisation rate, remote sensing at early stem elongation and the temperature sum during grain filling, was tested for two cultivars; Astoria (Secobra, France) and Wikingett (Svalöf-Weibulls, Sweden) in 16 fertilisation trials in southern Sweden, encompassing 3 years (2001–2003). Fertilisation was applied either as a single dose at sowing, or as both a starting application and an application at early stem elongation. The highest total application rate was 160 kg N ha⁻¹ y⁻¹. The soil was analysed for phosphorus, potassium, magnesium and calcium in the layer 0–0.3 m, and mineral N down to 0.6 m. Canopy reflectance observations at BBCH 32 was used to calculate a vegetation index (TCARI(32)) to reflect the canopy (leaf and straw) chlorophyll concentration. Harvested grain CP correlated only marginally with the observed soil variables. It was, however, for a specific cultivar, possible to make a prediction of grain CP based on day of sowing and TCARI(32) with . Part of the sowing day effect might be due to thermal stress during grain filling, as the risk for high temperatures during this phase was higher when sowing was late. This might also explain why the introduction of accumulated temperature during grain filling, into a model already including sowing day and TCARI(32) as independent variables, did not improve the predictability of grain CP.     

Effect of CHOP expression knock-down on apoptosis of renal tubular epithelial cells

AIM:To study the effect of C/EBP homologous protein (CHOP) on the apoptosis of renal tubular epithelial HK2 cells. METHODS:The serum mRNA levels of CHOP in the patients with acute kidney injury and healthy controls were detected by qPCR. In vitro, renal tubular epithelial HK2 cells were divided into control group, negative group (transfected with negative control siRNA), si-CHOP group (transfected with CHOP siRNA), and induced by transforming growth factor-β1 (TGF-β1). The viability of the cells was measured by MTT assay, and the apoptotic rate was analyzed by flow cytometry. The protein levels of nuclear antigen Ki-67, proliferating cell nuclear antigen (PCNA), caspase-3 and cleaved caspase-3 were determined by Western blot. RESULTS:Compared with the healthy controls, the serum mRNA levels of CHOP in the patients with acute kidney injury were increased significantly (P<0.05). Transfection with CHOP siRNA significantly decreased the expression of CHOP in the renal tubular epithelial HK2 cells (P<0.05). Knock-down of CHOP expression by siRNA significantly increased the viability of renal tubular epithelial HK2 cells (P<0.05), decreased the apoptotic rate (P<0.05), increased the expression of Ki-67 and PCNA (P<0.05), and down-regulated the protein level of cleaved caspase-3 (P<0.05). CONCLUSION:The serum mRNA levels of CHOP were increased in the patients with acute kidney injury. Knock-down of CHOP expression inhibits the apoptosis of renal tubular epithelial cells by regulating the expression of proliferation-and apoptosis-related proteins.     

PVX病毒载体介导2种马铃薯Y病毒科病毒VPg蛋白表达诱导烟草叶片系统坏死的研究

芜菁花叶病毒(Turnip mosaic virus,TuMV)和槟榔坏死环斑病毒(Areca plam necrotic ringspot virus,ANRSV)是马铃薯Y病毒科(Potyviridae)中分别可诱导十字花科作物和槟榔产生坏死病症的2种不同属病毒.VPg(Viral protein genome l...     

苏云金杆菌HBF-1菌株在土壤中毒蛋白含量的ELISA检测

以苏云金芽孢杆菌HBF-1菌株中分离纯化的高特异性杀虫蛋白作为抗原，免疫新西兰白兔制备出多克隆抗体，其效价可达到16000。应用制备的特异性抗体建立了间接酶联免疫吸附测定法（ELISA），检测不同土壤样品中Bt毒蛋白含量。初步结果表明，该方法能够检测土壤中Bt毒蛋白的含量，因土壤理化特性及组成成分的不同，检测到的毒蛋白含量也略有不同，河沙中的蛋白检测量最低，只有12．53％；另一种沙壤混合物中检测量达到76.16％。     

日粮能量和蛋白水平对瘦肉型生长育肥猪生长性能的影响

选取60±1.5 kg、90.0±1.5 kg杜长大(D×L×Y)生长育肥猪48头(公母比例为1∶1),通过对平均采食量(ADFI)、平均日增重(ADG)和料重比(F/G)进行比较分析,考察蛋白和能量水平对其生长性能的影响.结果表明:日粮能量和蛋白水平对60～90 kg瘦肉型生长育肥猪ADG、ADFI和F/G均有显著影响(P＜0.05);日粮能量和蛋白水平对90～120 kg生长育肥猪ADG的影响不显著(P＞0.05),对ADFI、F/G影响显著(P＜0.05).60～90 kg、90～120kg阶段杜长大生长育肥猪适宜的能量和蛋白水平分别为13.90 MJ·kg-1、15.90％和14.62 MJ'kg-1、14.90％;多元回归分析得到90～120 kg瘦肉型生长育肥猪DEI、CPI与ADG的回归方程为:△W=- 12.772DEI+ 23.467CPI+ 351.156(R2=0.881,P＜0.05).     

水稻瘤矮病毒髓和S8基因共表达杆状病毒转移载体构建及重组病毒的鉴定

为构建携带水稻瘤矮病毒（Rice gall dwarf virus,RGDV）主要内层衣壳蛋白53基因和外层衣壳蛋白鼹基因的重组杆状病毒,将目的基因（S3和S8）分别亚克隆到杆状病毒转移载体pFastBacDual多角体启动子（PH）和plO启动子的下游．经酶切和确证性序列测定,将其转化到DHl0Bac感受态细胞中,获得重组杆粒rbpFBDS3-S8,采用脂质体转染法,将rbpF-BDS3-58转染草地贪夜蛾（Spodoptera frugiperda）Si9细胞包装病毒,PCR筛选鉴定重组病毒．结果表明：SD昆虫细胞被侵染72h后,倒置显微镜下观察到细胞增大,培养液和细胞内出现颗粒状物质,部分细胞破裂甚至裂解,说明53和S8基因已整合到重组杆状病毒基因组中,这为开展RGDV主要结构蛋白在昆虫细胞中的表达及其功能研究奠定了基础．     

冷藏对郁金香鳞茎可溶性糖和蛋白质的影响

以郁金香鳞茎阿波罗、检阅、小黑人、普瑞斯玛、巴塞罗娜为试材,对不同冷藏处理时间的郁金香鳞茎不同部位的可溶性总糖、还原糖、蔗糖、可溶性蛋白质含量进行研究。结果表明:5个品种郁金香鳞茎在冷藏处理期间可溶性总糖、还原糖、蔗糖、可溶性蛋白质含量呈规律性变化。蔗糖是可溶性总糖的主要形式;初步认为解除休眠的过程是由外向内,通过鳞茎盘调节来实现的;5个品种郁金香鳞茎还原糖、蔗糖在第7周出现明显的波峰和拐点;检阅较其它4个品种表现出较高的可溶性糖含量和活性。     

半滑舌鳎(Cynoglossus semilaevis)雌性特异基因CSW3重组蛋白的表达与应用

CSW3基因序列来源于半滑舌鳎(Cynoglossus semilaevis)全基因组测序项目,是经过筛选得到的雌性特异候选基因之一。本研究探索了半滑舌鳎雌性相关基因CSW3的表达与应用,构建了半滑舌鳎雌性特异CSW3基因重组表达载体,通过大肠杆菌进行了体外重组表达,对基因表达产物进行分离纯化,并通过蛋白转染的方法研究了纯化的蛋白对几个性别相关基因的影响。结果显示,雌性特异CSW3重组蛋白注射鱼体后对半滑舌鳎性腺foxl2、sox9a、amh这3种性别相关基因的表达水平具有显著影响,能够引起雌性相关基因foxl2表达上调、雄性相关基因sox9a和amh表达下降。从蛋白水平对雌性相关基因CSW3的基因功能进行初步研究,为半滑舌鳎性别控制及全雌苗育种提供基因资源及技术方法。     

半滑舌鳎(Cynoglossus semilaevis) CSFR2基因的重组表达及其蛋白纯化与功能分析

通过对半滑舌鳎(Cynoglossus semilaevis)进行全基因组及转录组测序,预测了一个半滑舌鳎性别相关基因CSFR2,目前已对该基因进行了克隆和表达分析。为进一步研究CSFR2基因的功能,本研究从蛋白层面入手,构建了原核表达载体p ET-32a-CSFR2,转化到大肠杆菌E.coli进行诱导表达,用His Trap进行蛋白纯化,SDS-PAGE电泳检测诱导及纯化产物,并将表达纯化的CSFR2蛋白注射1龄半滑舌鳎精巢。由于CSFR2是一个性别相关基因,因此无法通过免疫反应来直接检测其蛋白活性,本研究通过不同时间点取样检测半滑舌鳎其他性别相关基因Cyp19a和Foxl2的表达量变化,来间接反应CSFR2蛋白的活性及功能。实验结果显示,重组蛋白能够进入到活体细胞内并发挥一定作用,在6–72 h内对性别相关基因Cyp19a和Foxl2的表达有上调作用,说明CSFR2蛋白具有生物活性,并能调节其他性别相关基因的表达量,间接促进雄激素转化为雌激素。本研究进一步了解CSFR2基因在半滑舌鳎性别决定与分化中的作用,可为半滑舌鳎性别控制提供理论依据,在人工诱导鱼类性逆转方面有重要应用价值。此外,本研究为蛋白活性验证提供了一种新方法。     

RNAi及沉默通路调控研究进展

RNA干扰（RNA interference,RNAi）是由双链RNA（double-stranded RNA,dsRNA）介导的细胞内双链mRNA特异性降解的现象,属于转录后的基因沉默机制。迄今为止,RNAi已经是一种广泛应用于研究基因功能的最新和最有效的方法,并且在治疗人类和动物疾病中也有很大潜力。作者主要综述了RNAi的过程、小RNA的生物发生学及沉默通路的保护和调控。