基于II型毒素–抗毒素系统构建ε–聚赖氨酸合成酶的链霉菌稳定表达载体

鉴于ε–聚赖氨酸収酵过程对抗生素压力条件的依赖,尝试基于Ⅱ型毒素–抗毒素系统(relBE2sca)构建无选择压力下稳定表达ε–聚赖氨酸的淀粉酶产色链霉菌表达系统:将抗毒素基因relB2sca与ε–聚赖氨酸合成酶基因pls克隆至表达载体,幵导入淀粉酶产色链霉菌pls缺失突变株,将毒素基因relE2sca整合至淀粉酶产色链霉菌的染色体(突变株YY3),获得包含ε–聚赖氨酸稳定表达的突变株YY1。经过多次传代,相比对照组,在不含抗生素压力条件下,突变株YY1依然能够稳定地合成ε–聚赖氨酸。毒素蛋白RelE2sca的表达会导致变铅青链霉菌、阿维链霉菌和链霉菌FR–008等常用链霉菌异源表达宿主的死亡,提示基于Ⅱ型毒素–抗毒素系统(relBE2sca)可作为一种通用的遗传标记。     

Genotypic differences in the antioxidant and carbon–nitrogen metabolism of acid-tolerant and acid-sensitive rice (Oryzasativa L.) cultivars under acid stress

A pot experiment was conducted to investigate differences in antioxidative defence and carbon–nitrogen metabolism between acid-tolerant (YJSM) and acid-sensitive (YHSM) rice cultivars under acid stress. Acid-tolerant and acid-sensitive rice were planted in both acidic soil (pH 4.21) and normal soil (pH 6.13). Forty-eight days after sowing, rice shoots and roots from four treatments were collected, and the other four replicates were harvested at seed maturity. The results revealed that the grain yield of acid-tolerant YJSM was significantly higher than that of acid-sensitive YHSM under acid stress. The activities of antioxidant enzymes (superoxide dismutase and catalase) and contents of non-enzymatic antioxidants (ascorbate and reduced glutathione) of acid-tolerant YJSM were both higher than those of acid-sensitive YHSM under acid stress. Moreover, the enzyme activities (nitrate reductase, glutamine synthetase, glutamate synthase and glutamate dehydrogenase) and product contents (soluble sugar and soluble protein) of carbon–nitrogen metabolism of acid-tolerant YJSM were higher than those of acid-sensitive YHSM under acid stress. The NO₃^–N and carbon (C) contents in leaves of acid-tolerant YJSM were both significantly higher than those of acid-sensitive YHSM under acid stress. This study suggests that the acid-tolerant rice cultivar has better antioxidative defence and carbon–nitrogen metabolism systems than the acid-sensitive rice cultivar and is more effective in resisting acid stress.     

Interactions between phosphorus availability and microbes in a wheat–maize double cropping system: A reduced fertilization scheme

Mechanisms controlling phosphorus(P) availability and the roles of microorganisms in the efficient utilization of soil P in the wheat–maize double cropping system are poorly understood. In the present study, we conducted a pot experiment for four consecutive wheat–maize seasons(2016–2018) using calcareous soils with high(30.36 mg kg^–1) and low(9.78 mg kg^–1) initial Olsen-P content to evaluate the effects of conventional P fertilizer application to both wheat and maize(Pwm) ...     

建鲤IL–1β和IL–8基因实时荧光定量RT–PCR检测方法的建立

根据Gen Bank上的基因序列,在保守区设定并合成特异性引物,选择β–actin作为内参基因,采用SYBR Green I染料法,进行熔解曲线分析和标准曲线的制作。熔解曲线表明,β–actin、IL–1β、IL–8的基因产物均为特异性产物,其Tm值分别为86、82.5和84℃;标准曲线表明,各基因Ct值的检测范围为12~32,扩增效率分别为96.3%,103.2%和102.6%,具有良好的线性关系,且r2均大于0.990;组内变异系数分别为0.14%~0.86%,0.18%~0.93%和0.13%~0.86%;用建立的方法检测健康建鲤头、肾组织中IL–1β、IL–8的表达情况,相对表达量分别为1.09和1.71。综合分析,建立的建鲤IL–1β、IL–8基因实时荧光定量PCR方法具有检测范围广,扩增效率高,特异性强,重复性高的特点,可用于建鲤IL–1β、IL–8基因的表达测定。     

外槽轮排肥器排肥离散元仿真及排肥舌参数优化

为通过优化外槽轮排肥器排肥舌结构,以提升其排肥均匀性,利用EDEM建立外槽轮排肥器的离散元仿真模型,对其排肥作业过程进行模拟仿真,通过肥料颗粒运动状态仿真结果验证肥料颗粒流动特性。应用虚拟试验方法,分析排肥舌倒角结构参数变化对肥料颗粒流动特性的影响。结果表明:排肥流量变异系数随着排肥舌倒角的增大而先减小后增大,两者间呈负抛物线型二次函数关系,且在排肥舌倒角弧长与单个凹槽弧长比值为0.85时,外槽轮排肥器排肥质量波动最小,排肥均匀性最佳。验证试验结果表明:配装最优结构排肥舌的外槽轮排肥器排肥流量变异系数最小,具有最佳的排肥均匀性,比传统平端排肥舌排肥流量均匀性提高了49.9%。     

Effect of Continuous Rice–Wheat Rotation on Soil Properties from Four Agro‐ecosystems of Indian Punjab

In Indian Punjab, rice–wheat is a dominant cropping system in four agro‐ecosystems, namely undulating subregion (zone 1), Piedmont alluvial plains (zone 2), central alluvial plains (zone 3), and southwestern alluvial plains (zone 4), varying in rainfall and temperature. Static and temporal variabilities in soil physical and chemical properties prevail because of alluvial parent material, management/tillage operations, and duration of rice–wheat rotation. A detailed survey was undertaken to study the long‐term effect of rice–wheat rotation on soil physical (soil separates, bulk density, modulus of rupture, saturated and unsaturated hydraulic conductivities, soil water content, and suction relations) and chemical (organic carbon, pH, electrical conductivity) properties of different textured soils (sandy clay loam, loam, clay loam, and silty clay loam) in these four zones of Punjab. Soil samples (of 0‐ to 30‐cm depth) from 45 sites were collected during 2006 and were analyzed for physical and chemical properties. The results showed that sand content and pH increased whereas silt and organic carbon decreased significantly from zones 1 to 4. Compared to other textures, significantly greater organic carbon, modulus of rupture, and pH in silty clay loam; greater bulk density in clay loam, and greater saturated hydraulic conductivity in sandy clay loam were observed. Irrespective of zone and soil texture, in the subsurface soil, there was a hard pan at 15–22.5 cm deep, which had high soil bulk density, modulus of rupture, more silt and clay contents (by 3–5%) and less organic carbon and hydraulic conductivity than the surface (0–15 cm) layer. These properties deteriorated with fineness of the soil texture and less organic carbon content. Continuous rice–wheat cropping had a deleterious effect on many soil properties. Many of these soils would benefit from the addition of organic matter, and crop yields may also be affected by the distinct hardpan that exists between 15 and 22.5 cm deep.