一种高效制备慢病毒表达载体的方法

慢病毒表达载体具有感染宿主广和基因组整合效率高的优点,因此在转基因动物的制备中得到广泛应用.本研究构建了含有红色荧光和绿色荧光的慢病毒表达载体,用磷酸钙转染法将三质粒慢病毒载体转染293T细胞,转染效率达72%,降低了慢病毒的生产成本.在进行慢病毒的浓缩与纯化时,通过超速度离心和超滤相结合的方法,在150 mL的慢病毒原液中经纯化后得到80 μL效价为2.38×109 TU/mL的慢病毒颗粒.获得的高效价双荧光慢病毒表达载体可为后续双基因转基因动物的制备提供可靠的制作途径与检测基础.     

Anti-stress effects of Glycine tomentella Hayata in tilapia: inhibiting COX-2 expression and enhancing EPA synthesis in erythrocyte membrane and fish growth

The objective of this study was to elucidate the in vivo effects of the ethanol extract of wooly Glycine tomentella Hayata (GTE) root on tilapia to elucidate whether GTE has antistress activity. Tilapia as an animal model were fed with or without GTE, then injected with lipopolysaccharide (LPS) or ammonium chloride (NH(4)Cl). The tilapia were exposed to 100 mg/L of aqueous NH(4)Cl, and/or acute cold stress. Growth parameters of the tilapia were measured during the feeding trials. Tilapia injected with GTE (20 μg/g of fish), NH(4)Cl (100 μg/g of fish) and/or LPS (1 μg/g of fish) were then sampled 2 h poststimulation. GTE significantly inhibited cyclooxygenase-2 expression and hemoglobin (Hb) dimer formation (36 kDa). GTE also improved growth and blood viscosity and upregulated eicosapentaenoic acid content of erythrocytes. The in vivo results indicated that GTE (20 μg/g of fish) can be applied as a stress-tolerance enhancing agent for the aquaculture industry.     

Multiresidue analysis of 58 pesticides in bean products by disposable pipet extraction (DPX) cleanup and gas chromatography-mass spectrometry determination

A method based on disposable pipet extraction (DPX) sample cleanup and gas chromatography with mass spectrometric detection by selected ion monitoring (GC/MS-SIM) was established for 58 targeted pesticide residues in soybean, mung bean, adzuki bean and black bean. Samples were extracted with acetonitrile and concentrated (nitrogen gas flow) prior to being aspirated into DPX tubes. Cleanup procedure was achieved in a simple DPX-Qg tube. Matrix-matched calibrations were analyzed, and the limits of quantification (LOQ) of this method ranged from 0.01 mg kg(-1) to 0.1 mg kg(-1) for all target compounds. Coefficients of determination of the linear ranges were between 0.9919 and 0.9998. Recoveries of fortified level 0.02 mg kg(-1) on soybean, mung bean, adzuki bean and black bean were 70.2-109.6%, 69.1-119.0%, 69.1-119.8%, and 69.0-120.8%, respectively, for all studied pesticides. Moreover, pesticide risk assessment for all the detected residues in 178 market samples at Beijing market area was conducted. A maximum 0.958% of ADI (acceptable daily intake) for NESDI (national estimated daily intake) and 55.1% of ARfD (acute reference dose) for NESTI (national estimated short-term intake) indicated low diet risk of these products.     

Determination of multiresidues in rapeseed, rapeseed oil, and rapeseed meal by acetonitrile extraction, low-temperature cleanup, and detection by liquid chromatography with tandem mass spectrometry

A multiresidue method for determining pesticides in rapeseed, rapeseed oil, and rapeseed meal by use of liquid chromatography-tandem mass spectrometry is developed. Samples were extracted with acetonitrile or acidified acetonitrile and cleaned up by a 12 h freezing step. The recovery data were obtained by spiking blank samples at three concentration levels. The recoveries of 27 selected pesticides in rapeseed, rapeseed oil, and rapeseed meal were in the range of 70-118%, at the concentration level of 10 μg kg(-1), with intraday and interday precisions of lower than 22 and 27%, respectively. Linearity was studied between 2 and 500 μg L(-1) with determination coefficients (R(2)) of higher than 0.98 for all compounds in the three matrices. The limits of quantitation (LOQs) of pesticides in rapeseed, rapeseed oil, and rapeseed meal ranged from 0.3 to 18 μg kg(-1). The n-octanol-water partition coefficient showed more influence than water solubility in extracting pesticides by acetonitrile from matrices of high fat content. This method was successfully applied for routine analysis in commercial products.     

油菜籽粒点面接触碰撞中恢复系数的测定及分析

为建立油菜籽粒与播种、收获等机具相关重要工作部件间发生碰撞时的碰撞模型,该文基于运动学方程构造了油菜籽粒恢复系数的测定装置,并针对油菜籽粒的弹性特性进行了试验测定和分析。针对华双4号油菜品种,采用L8(41×24)混合正交试验方案研究了碰撞材料、碰撞材料厚度、自由下落高度和含水率等因素对油菜籽粒恢复系数的影响,然后针对碰撞材料、下落高度和碰撞材料厚度等重要因素进行了单因素试验,并进一步获得下落高度和碰撞材料厚度2个因素对恢复系数的影响规律及回归方程。试验结果表明,影响油菜籽粒恢复系数的因素主次顺序为:碰撞材料、自由下落高度、碰撞材料厚度和含水率。油菜籽粒与Q235、铝合金、有机玻璃、橡胶等碰撞材料之间恢复系数依次减小,并且随着下落高度的增加而减小,随着碰撞材料厚度的增加而增大。在碰撞材料为厚1 mm的Q235和籽粒含水率8.06%条件下,恢复系数随下落高度变化的回归方程决定系数R2=0.9979;在碰撞材料为Q235、籽粒含水率8.06%和下落高度420 mm条件下,恢复系数随材料厚度变化的回归方程决定系数R2=0.9872。该文结果可为油菜精密播种及高性能收获机具中相关工作部件的设计提供依据。     

Soil nitrogen availability alters rhizodeposition carbon flux into the soil microbial community

Purpose

Soil microorganisms are important in the cycling of plant nutrients. Soil microbial biomass, community structure, and activity are mainly affected by carbon substrate and nutrient availability. The objective was to test if both the overall soil microbial community structure and the community-utilizing plant-derived carbon entering the soil as rhizodeposition were affected by soil carbon (C) and nitrogen (N) availability.

Materials and methods

A ¹³C-CO₂ steady-state labeling experiment was conducted in a ryegrass system. Four soil treatments were established: control, amendment with carboxymethyl cellulose (CMC), amendment with ammonium nitrate (NF), combined CMC and NF. Soil phospholipid fatty acid (PLFA) and ¹³C labeling PLFA were extracted and detected by isotope ratio mass spectrometer.

Results and discussion

The combined CMC and NF treatment with appropriate C/N ratio (20) significantly enhanced soil microbial biomass C and N, but resulted in lower soil inorganic N concentrations. There was no significant difference in soil PLFA profile pattern between different treatments. In contrast, most of the ¹³C was distributed into PLFAs 18:2ω6,9c, 18:1ω7c, and 18:1ω9c, indicative of fungi and gram-negative bacteria. The inorganic-only treatment was distinct in ¹³C PLFA pattern from the other treatments in the first period of labeling. Factor loadings of individual PLFAs confirmed that gram-positive bacteria had relatively greater plant-derived C contents in the inorganic-only treatment, but fungi were more enriched in the other treatments.

Conclusions

Amendments with CMC can improve N transformation processes, and the ryegrass rhizodeposition carbon flux into the soil microbial community is strongly modified by soil N availability.

     

引入地形因子的黑土区大豆干生物量遥感反演模型及验证

为了对田块尺度农作物地上干生物量进行估测,提高大豆地上干生物量反演模型的精度和稳定性,该文获取了研究区地块2016年7、8月份的SPOT-6多光谱数据,并测定不同地形坡位的大豆地上干生物量,以归一化植被指数(normalized difference vegetation index,NDVI)和增强型植被指数(enhanced vegetation index,EVI)为输入量,建立田块尺度大豆地上干生物量一元线性回归模型;加入与地上干生物量相关的地形因子,建立逐步多元回归和神经网络多层感知反演模型.结果表明:1)使用传统的单一植被指数模型预测大豆地上干生物量有可行性,但模型精度和稳定性不高.2)加入地形因子(海拔、坡度、坡向)的神经网络多层感知器模型,有较高的精度和可靠性,模型准确度达到90.4％,验证结果显示预估精度为96.2％.反演结果与地块的地形、地貌、气温和降水特征基本吻合,反映了作物长势的空间分布特征,可以为田块尺度大豆地上干生物量动态监测和精准管理,提供借科学依据.     

马铃薯精密播种机智能控制系统设计

针对现有马铃薯播种机播种株距控制精准度不高、易产生重种漏种等问题,研发了一种由主控制模块、检测模块、株距控制模块和振动强度控制模块等7个模块组成的马铃薯精密播种机智能控制系统,采用液压马达控制薯种输送带运转,步进电机控制薯种输送带的振动强度,实现了播种株距和重种漏种率的自动控制.试验结果表明,播种速度相同时,实际播种株距相对于设定播种株距的平均偏差依次增大,播种速度越高实际播种株距的稳定性越差;薯种输送带振动强度越强,重种率越低,漏种率越高,各因素对重种漏种率影响的主次顺序为:薯种输送带振动强度＞播种速度＞薯种质量,且薯种输送带振动强度对重种率、薯种输送带振动强度和播种速度对漏种率有显著影响;较佳的播种作业参数为:薯种输送带振动强度为Ⅱ级(即轻微振动时)、播种速度为1.16 m/s及薯种质量为35 g.经2～3个周期即可调整到允许范围内,且稳定性好.因此,完全能够满足种植户的实际播种作业要求,为智能控制马铃薯精密播种装备的后续研发提供参考.     

用核磁共振研究浸种方法对水稻种子吸水量的影响

为寻求较佳浸种方法,该文应用低场核磁共振检测技术,研究了不同的浸种方式及浸种溶剂对水稻种子吸水量的影响。试验利用横向弛豫时间 T2反演谱分析了水稻种子的水分状态变化及吸水特性,发现浸种过程改变了水稻种子内部的水分分布情况,水稻种子吸水量对初始含水率差异不显著(P>0.05),但对各种浸种方法差异显著(P<0.05)。研究表明,采用连续浸种4 h、浸种3 h-晾干1 h-浸种1 h、浸种2 h-晾干1 h-浸种2 h及浸种2 h-晾干2 h-浸种2 h这4种不同的浸种方式时,浸种2 h-晾干1 h-浸种2 h的间歇浸种方式吸水率较高;采用清水、强氯精300倍液、饱和澄清石灰水、质量分数为40%福尔马林的50倍液、100倍液及200倍液6种不同的浸种溶液时,应用质量分数为40%福尔马林50倍液药剂时吸水率较高。低场核磁共振检测技术揭示了水稻种子含水量的影响因素,为浸种过程中吸水量的测定提供了一种有效的方法。     

心土培肥犁改良瘠薄土壤的效果

研究根据心土培肥的改土技术要求研制出心土培肥犁,并分别在瘠薄黑土和碳酸盐草甸黑钙土上开展大面积机械改土试验,明确自主研发的心土培肥犁改土后对土壤理化性质影响及对作物产量的效果,为其广泛应用到低产土壤改良提供机械及技术支持。试验设深松、心土培肥和常规对照耕作,采用大田对比方法。研究结果表明:心土培肥和深松在不同类型土壤上对土壤理、化性质,对作物产量及产量性状影响后效不完全一致;心土培肥降低土壤抗剪强度后效明显,碳酸盐草甸黑钙土10~30 cm土层土壤抗剪强度比对照降低6.65~12.16 k Pa,黑土比对照降低8.20~11.31 k Pa,碳酸盐草甸黑钙土改土后效果明显,黑土改土后效长,心土培肥改土效果优于深松;土壤容质量和硬度趋势同上;心土培肥提高土壤透气系数为2.78~14.28倍,饱和导水率为2.38~11.62倍;深松和心土培肥可提高下层土水分消耗比例,30~60 cm土层耗水量为心土培肥区深松区对照区,心土培肥耗水量比照高10%;心土培肥处理可提高土壤磷含量和供磷强度,20~30 cm和30~40 cm土层土壤供磷强度比对照分别提高4.19~5.17倍和4.96~17倍,碳酸盐草甸黑钙土高于黑土;心土培肥可提高玉米产量,碳酸盐草甸黑钙土上心土培肥增产幅度为6.82%~18.01%,黑土增产幅度为6.45%~11.18%,平均增产效果碳酸盐草甸黑钙土薄层黑土,但黑土持续增产效果好。