GFP与水稻条纹病毒病害特异蛋白的融合基因在sf9昆虫细胞中的表达

 用重叠延伸PCR(overlap Polymerase Chain Reaction,overlap-PCR)方法获得了含有绿色荧光蛋白(Green fluorescence protein,GFP)和水稻条纹病毒(Rice stripe virus,RSV)病害特异性蛋白SP(Disease-specific protein)两者的融合基因(GFP-SP),并将其克隆至载体pMD18-T,得到的重组质粒pMD18-T-GFP-SP经Xba Ⅰ/Hind Ⅲ双酶切后与经相同方法处理过的杆状病毒转移载体pFastBacHTb相连接构建成重组转移质粒pFastBacHTb-GFP-SP。酶切和测序鉴定证明了其序列的正确性并且无移码现象发生。将此质粒转化入含穿梭载体Bacmid的感受态细胞DH10Bac,得到含有目的基因片段的重组杆状病毒穿梭质粒rb-GFP-SP。以之转染草地贪夜蛾(Spodoptera frugiperda,sf9)细胞,24~48h后在显微镜200倍可见光视野下观察到被感染细胞发生了细胞和细胞核变大、细胞内颗粒物增多、细胞脱落甚至裂解等一系列与正常的st9昆虫细胞形态有明显区别的变化。荧光显微镜下可见部分细胞发出清晰的绿色荧光,且大部分荧光集中在细胞质部分。这些结果表明,GFP-SP融合蛋白在st9昆虫细胞内成功表达。     

侵染葫芦的黄瓜绿斑驳花叶病毒广西分离物分子鉴定

从广西南宁市郊温室大棚中的葫芦[Lagenaria siceraria(Molina)Stand.]上采集到一个表现脉绿、花叶症状的病毒样品,ELISA检测表明,该样品与黄瓜绿斑驳花叶病毒(Cucumber green mottle mosaic virus,CGMMV)有密切的血清学关系,利用RT-PCR方法从样品中扩增获得约500bp的DNA片段,序列分析表明,该片段是CGMMV的外壳蛋白基因,暂将该病毒分离物定名为GX-BG。外壳蛋白基因核苷酸序列系统进化树分析表明,已报道的CGMMV主要分为3大群体,GX-BG与中国辽宁分离物(CGMMV-LN)分别属于不同的群体。     

Antibacterial activity in microalgae cultures

Five cultures of microalgae (Chlorella minutissima, Tetraselmis chui, Nannochloropsis sp., Arthrospira platensis and Isochrysis sp.) with no culturable bacteria were tested for their ability to inhibit the growth of six Vibrio bacterial strains (V. parahaemolyticus, V. anguillarum, V. splendidus, V. scophthalmi, V. alginolyticus and V. lentus). The influence of light on the antibacterial activity of the microalgae was investigated. All microalgae cultures inhibited the growth of bacteria compared with the control treatments (P < 0.05), and their antibacterial activity was not influenced by the presence or absence of light. In the control groups, the numbers of bacteria increased exponentially during the experimental period in the absence of microalgae cells demonstrating that the bacterial cells were able to utilize the growth medium of microalgae cultures. The present results may explain the low levels or absence of Vibrio strains in microalgae cultures, and the positive effect of addition of microalgae in rearing of fish larvae, and implicate the production of antibacterial compounds by microalgae cells.     

Effect of salinity on growth and energy budget of red and green colour variant sea cucumber Apostichopus japonicus (Selenca)

Sea cucumber, Apostichopus japonicus (Selenca), tolerates salinity fluctuations inhabiting intertide zone. This study deals with growth, food intake, food conversion and the bioenergetic responses of the red variant (wet weight of 2.60 ± 0.11g) and green variant (wet weight of 2.56 ± 0.08 g) A. japonicus to different salinities of 22, 26, 30, 34, and 38 psu at 16.5 ± 0.5°C. The results showed that salinity had a significant effect on specific growth rate (SGR) of both green and red variants A. japonicus (P < 0.05). Both colour variants of sea cucumber had highest SGR at 30 psu, and then decreased when salinity below or above this point. Maximum SGR (the green 1.07 ± 0.08% day^?1, the red 1.14 ± 0.09% day^?1 respectively) is related with maximum food intake (FI) and highest food conversion efficiency (FCE) (P < 0.05) occurring at 30 psu. Only under 22 psu, the green variant grew faster than the red variant (P < 0.05), and under other four salinity treatments there was no significant difference between SGR of two colour variant holothruians (P > 0.05). Values of adaptable salinity scope for green and red variants sea cucumber survival are 18.5~39 psu and 20.9~38.6 psu respectively. The average energy budget formula of sea cucumber at 30 psu was: 100C = 6G +42F +3U+49R (C, energy ingested; G, energy for growth; F, energy loss as faeces; U, energy used for ammonia excretion; R, energy loss for respiration). The sea cucumber had maximum energy ingested (C) and highest proportion of energy for growth (G) at 30 psu, and then decreased when salinity is above or below this salinity. Both red and green variants of A. japonicus deposited for growth were very low, and the energy loss in faeces and energy for respiration accounted for the majority of assimilation energy. The result clearly showed that the optimum condition for farming green and red variants A. japonicus, both with respect growth and energy allocation, is the salinity scope of 26 ~ 30 psu.     

Growth and survival of the juvenile yellowleg shrimp Farfantepenaeus californiensis cohabiting with the green feather alga Caulerpa sertularioides at different temperatures

This study was performed to determine the growth and survival patterns of the juvenile yellowleg shrimp Farfantepenaeus californiensis cohabiting with the green feather alga Caulerpa sertularioides over a range of temperatures (18, 22, 25, 28 and 32°C) found throughout the year in the Gulf of California. From an initial weight of 46 ± 10 mg, shrimp cohabiting with green feather algae increased in weight by an average of 0.57 (±0.07), 5.44 (±0.45), 6.36 (±0.37), 7.66 (±0.26) and 5.96 (±0.59) g at 18, 22, 25, 28 and 32°C, respectively, whereas shrimp grown without seaweed increased in weight by an average of 0.28 (±0.026), 2.66 (±0.24), 3.15 (±0.49), 4.00 (±0.67), and 3.54 (±0.09) g respectively. Statistically significant differences were observed for the growth rates of juvenile yellowleg shrimp in the presence of C. sertularioides and peak growth was observed at 28°C in experiments with macroalgae. No significant differences were observed for shrimp survival except at 32°C without macroalgae, where survival was lower. These results suggest a beneficial and growth‐promoting effect of cohabitation with live C. sertularioides on F. californiensis.     

蟠桃果实总RNA提取方法的研究

[目的]筛选获得提取蟠桃果肉中总RNA的适宜方法.[方法]以盛花后100 d的蟠桃为材料,采用Transplant plus植物总RNA提取试剂提取法、RNAisoTM试剂盒法、改良SDS法、改良CTAB法等四种方法提取总RNA.[结果]两种试剂盒法提取的总RNA质量差,有严重的基因组污染;改良SDS法提取物中含有大量的多糖;采用改良CTAB法所得RNA完整性好,条带清晰无降解,无基因组污染.经过RT - PCR和Northern blot检测后,表明该RNA能够满足后续分子生物学操作的要求.[结论]蟠桃果实总RNA适宜提取方法-改良CTAB法的确定为后续果实发育分子生物学的研究奠定了基础.     

蟠桃采后贮藏前后香气成分变化的研究

[目的]分析研究采后新疆蟠桃在贮藏前后期果实香气成分的变化.[方法]采用顶空-固相微萃取( HS - SPME)与气-质联用(GC - MS)技术对蟠桃果实香气成分进行提取和分析;比较溶剂萃取(LLE)、水蒸气蒸馏(SDE)和HS - SPME 3种提取方法对蟠桃香气成分分析的影响.[结果]HS - SPME最佳萃取条件为萃取头50/30μm DVB/CAR/PDMS灰色,萃取时间40 min,萃取温度50℃;HS - SPME - GC - MS在蟠桃果实贮藏前后期分别检测出53和44种挥发性物质.[结论]贮藏前后期蟠桃果实所含的挥发性成分有较大的差异,作为蟠桃香气的特征组分酯类和内酯类物质在贮藏第4周后含量显著减少.     

基于城市文化资源配置与表达的绿地系统规划方法初探——以石家庄栾城县为例

在规划石家庄市栾城县绿地系统的过程中,以布局上体现城市文化特色为主要切入点,从城市-区域-个体绿地的宏、中、微观分析出发,注重宏观上的合理分配,中观上的有机结合,微观上的个性体现,将有特色的城市文化合理分配,创造了系统完整的,区域分明的,特色各异的文化型绿地,并在绿地系统规划的过程中辅助了绿地的有机布局与特色表达,赋予城市绿地更多文化定位和内涵,塑造了多功能,多样性的文化型绿地形式。     

Effects of Chemical Components on the Amount of Green Tea Cream

The relationship between the amount of tea cream and the chemical components contents in the original green tea infusion was investigated. The results showed that the amounts of tea cream in the green tea infusions obtained from different cultivars and different parts of new shoots were varied. There were many chemical components participating in the formation of green tea cream. However, there were only the contents of caffeine (Y=0.85, P＜0.01) and polyphenols (Y=0.65,P＜0.05) in the original green tea infusion highly correlated with the amount of green tea cream. Stepwise regression analysis of overall chemical components indicated that the contents of caffeine and gallated catechins in the original green tea infusion had a significant effect (P＜0.0 1) on green tea cream levels. Cream (g L-1)=-172.071+ 0.129×Ccaffeine+0.024×Ggallated catechins (R2=0.936). The amount of green tea cream can be predicted by the contents of gallated catechins and caffeine in the original tea infusion. Principal component analysis also indicated that catechins, minerals, and polysaccharides were the important chemical components in the formation of green tea cream.     

基于模型的上海郊区地下水氮素非点源污染特征研究

农业非点源污染是造成上海郊区地下水污染的主要因素,定量分析预测农业生产过程氮素的迁移转化规律是有效控制地下水污染的重要环节。以上海市浦东新区新场镇果园村的桃园为研究对象,借助生物地球化学过程模型(DNDC)和长期水文影响评价模型(L-THIA),基于连续观测数据,详细分析了农业生产过程中氮素造成的非点源污染,特别是对周边地表、地下水的影响。结果表明,地表水总氮均值达6.34mg·L-1,远劣于地表水Ⅴ类标准(≤2.0mg·L-1);地下水中总氮均值达16.85mg·L-1,远劣于地表水Ⅴ类标准(≤2.0mg·L-1)。约有20%采样点硝态氮含量属于地下水Ⅴ类(>30mg·L-1)。野外检测数据表明,该区地表水、地下水污染均严重超标,不宜饮用。模型分析显示,水体污染源主要来自桃园生产中施用的肥料,其中就模拟结果的数值可以得出,大约年农田氮输入量的1.7%通过土壤径流进入地表水,约3.5%经过土壤渗漏进入地下水,实测地下水中氮含量占桃园总氮输入量的5.8%。因此,合理调整施肥措施和施肥结构是减少土壤-水体中氮素污染的有效途径。