PiggyBac转座子介导的诱导细胞永生化转基因载体的构建及其整合效率的检测

【目的】构建在小鼠水平上实现多个目的基因的表达以及标记基因安全删除的转基因载体。【方法】以载体为模板,分别扩增SV40P neo、IRES、tk-PloyA,通过overlap PCR连接,并在两端添加方向相同的LoxP序列,构建转基因基本载体PB-NIT;然后通过overlap PCR扩增获得Tet-CMV-SV40 T-T2A-p 53-PolyA基因表达盒,将其插入PB-NIT中,构建载体PB-NIT-STP;最后将转录因子激活域 rtTA 通过同尾酶连接插入PB-NIT-STP,构建载体PB-rtTA-NIT-STP。【结果】经酶切和测序等鉴定,以上载体均正确;经转座活性鉴定,共转染转座子载体PB-rtTA-NIT-STP和转座酶载体比只转染转座子载体获得的阳性克隆数提高了20倍。【结论】得到了基于PiggyBac转座子的可用于正负向筛选和诱导目的基因表达的转基因载体。     

不同种类有机物对污染农田土壤重金属活性的影响

为了解施用不同种类有机物对污染农田土壤重金属行为的影响,布设了田间小区试验,观察研究施用6种不同类型有机物对土壤水溶性重金属和农作物对重金属吸收等的影响。试验设7个处理,分别为对照(不施有机肥)、施新鲜水稻秸秆、新鲜鸡粪、鸡粪堆肥、新鲜猪粪、猪粪堆肥和商品有机肥,动态检测土壤中水溶性重金属含量、田面水中重金属浓度及收获水稻籽粒中重金属含量的变化。结果表明,与不施有机物的对照处理比较,施用各类有机物在试验前期均可明显提高土壤中水溶性重金属和田面水中重金属的含量,总体上以施用新鲜猪粪的最高,其次为新鲜鸡粪,施用商品有机肥的最低。土壤中水溶性重金属含量随试验时间增长逐渐下降。施用各类有机物对水稻籽粒中重金属的积累影响不明显。土壤中水溶性重金属及田面水中重金属含量均与相应的水溶性碳(DOC)浓度呈显著相关性,表明因有机物施用增加土壤中DOC浓度可能是土壤中水溶性重金属含量增加的主要原因;而不同有机物对土壤重金属活性影响的差异可能与有机物本身的分解程度及可释放DOC大小有关。但施用有机物引起的农田土壤水溶性重金属含量的增加是暂时的,对水稻生长后期籽粒中重金属积累的影响不明显。     

Physical activation of biochar and its meaning for soil fertility and nutrient leaching – a greenhouse experiment

The slow alteration of the surface of charred biomass (biochar) over time may contribute to an improved nutrient retention and thus fertility of tropical soils. Here, we investigated soils from temperate climates and investigated whether a technical steam activation of biochar could accelerate its positive effects on nutrient retention and uptake by plants relative to nonactivated biochar. To this aim, we performed microcosm experiments with sandy or silty soil, mixed with 2.0, 7.5 and 15.0 g/kg soil of fine (<2 mm) or coarse‐sized (2–10 mm) biochar from beech wood (Fagus sp.). After initial fertilizer (NPK), ashes and excess nutrients were leached with water, and the microcosms were planted for 142 days with Italian Ryegrass (Lolium multiflorum ssp. italicum). Thereafter, leachate, soil and plant samples were analysed for their nutrient contents. The results showed that biochar additions of ≤15 g/kg soil left elevated contents of available P and N in the surface soil but reduced their uptake into the plants. As a result, total biomass production was unchanged. Different particle size and application amounts influenced these findings only marginally. Nitrate leaching was enhanced in the sandy soil (+41% for nitrate, but reduced in the silty soil ?17%) and P was immobilized. Hence, the fertility of the temperate soils under study was only marginally affected by pure biochar amendments. Steam activation, however, almost doubled the positive effects of biochars in all instances, thus being an interesting option for future biochar applications.     

水稻OsZ314基因克隆及转录激活验证

穗是水稻(Oryza sativa L.)重要的生殖器官,其生长发育直接影响水稻的产量和品质。前期研究中,我们对水稻生殖生长时期穗发育各阶段的转录组、蛋白组数据进行联合分析,筛选到一个MYB基因家族的转录因子OsZ314基因。生物学信息分析表明,该基因位于1号染色体上,拥有典型的R2R3-MYB型转录因子结构域,且在水稻幼穗时期表达量最高;亚细胞定位结果显示,OsZ314基因定位于细胞核中;转录激活试验表明,OsZ314基因具有转录激活功能。本研究为深入探索OsZ314基因在水稻穗发育时期的分子生物学功能奠定了良好的基础。     

Experiment on Activation of Black Soil by Electro-hydraulic Impulse

The activation of black soil (analog the lignite) by electro-hydraulic impulse is proposed. The abundant insoluble nitrogen, phosphorus, potassium and other organic elements in black soil are activated as the green fertilizer by this way. The new method is provided for activating sludge produced by treating wastewater. For several kinds of black soil, the experiment result indicates that electro-hydraulic impulse increases the percentage of absorbable nitrogen by 1 to 4 times and the percentage of insoluble organic elements by 4 times, which is a valuable reference to sludge treatment.     

No effect of exogenous melatonin on development of cryopreserved metaphase II oocytes in mouse

Background

This study was conducted to investigate effect of exogenous melatonin on the development of mouse mature oocytes after cryopreservation.

Results

First, mouse metaphase II (MII) oocytes were vitrified in the open-pulled straws (OPS). After warming, they were cultured for 1 h in M₂ medium containing melatonin at different concentrations (0, 10⁻⁹, 10⁻⁷, 10⁻⁵, 10⁻³ mol/L). Then the oocytes were used to detect reactive oxygen species (ROS) and glutathione (GSH) levels (fluorescence microscopy), and the developmental potential after parthenogenetic activation. The experimental results showed that the ROS level and cleavage rate in 10⁻³ mol/L melatonin group was significantly lower than that in melatonin-free group (control). The GSH levels and blastocyst rates in all melatonin-treated groups were similar to that in control. Based on the above results, we detected the expression of gene Hsp90aa1, Hsf1, Hspa1b, Nrf2 and Bcl-x1 with qRT-PCR in oocytes treated with 10⁻⁷, or 10⁻³ mol/L melatonin and untreated control. After warming and culture for 1 h, the oocytes showed higher Hsp90aa1 expression in 10⁻⁷ mol/L melatonin-treated group than in the control (P < 0.05); the Hsf1, Hsp90aa1 and Bcl-x1 expression were significantly decreased in 10⁻³ mol/L melatonin-treated group when compared to the control. Based on the above results and previous research, we detected the development of vitrified-warmed oocytes treated with either 10⁻⁷ or 0 mol/L melatonin by in vitro fertilization. No difference was observed between them.

Conclusions

Our results indicate that the supplementation of melatonin (10⁻⁹ to 10⁻³ mol/L) in culture medium and incubation for 1 h did not improve the subsequent developmental potential of vitrified-warmed mouse MII oocytes, even if there were alteration in gene expression.     

温度对草鱼、鲤、鲢、鳙主要消化酶活性的影响

草鱼、鲤、鲢、鳙肝胰脏和肠蛋白酶最适温度分别为:45℃,45℃,55℃,55℃和45℃,45℃,50℃,50℃。临界失效温度分别为:56℃,54℃,62℃,61℃和54℃,57℃,61℃,60℃。淀粉酶最适温度分别为:30℃,30℃,35℃,35℃和30℃,30℃,35℃,35℃。临界失效温度分别为:53℃,46℃,52℃,49℃和50℃,47℃,50℃,52℃。脂肪酶最适温度分别为:30℃,30℃,25℃,25℃和30℃,30℃,30℃,30℃。临界失效温度分别为:45℃,38℃,38℃,38℃和45℃,38℃,38℃,38℃和42℃,40℃,40℃,40℃。草鱼、鲤肝胰脏的蛋白酶、淀粉酶、脂肪酶在最适温度条件下的活化能比肠的低,而鲢、鳙肝胰脏的蛋白酶、淀粉酶、脂肪酶最适温度条件下的活化能则比肠高。四种鱼最适温度下肝胰脏蛋白酶活化能的顺序为:鲤<草鱼<鲢<鳙;淀粉酶活化能的顺序为:草鱼<鲤<鲢<鳙;脂肪酶活化能的顺序为:鲤<草鱼<鲢<鳙。肠蛋白酶活化能的顺序为:鲤<鲢<鳙<草鱼;肠淀粉酶活化能的顺序为:草鱼<鲤<鲢<鳙;肠脂肪酶活化能的顺序为:鳙<鲢<草鱼<鲤。说明消化酶的活性与酶本身特性之一活化能高低有直接关系。     

农田沙漠化演变中土壤质量的生物学特性变化

通过研究处于沙漠化过程中的科尔沁沙地旱作农田土壤酶活性与基础土壤呼吸活性，分析了土壤质量生物学特性的沙漠化演变。结果表明，土壤酶活性与基础土壤呼吸对沙漠过程中环境的变化反应敏感，随着沙漠化程度增加，与土壤物理化学肥力因素有关的过氧化氢酶、脲酶和碱性磷酸酶的活性和土壤呼吸活性随之下降。土壤酶活性和呼吸活性与土壤有机质和养分有极显著的相关，在一定程度上可以用酶活性和呼吸强度表征有机质和养分的状况及其数量的变异，因此，可以作为农田沙漠化演变中土壤质量特征变化的重要辅助指标。     

脉冲次数和卵龄对小鼠卵母细胞电活化效果的影响

以一定电场条件、不同次数的电脉冲刺激注射ＨＣＧ后不同时间（１５、１７、１９、２１和２５ｈ）小鼠卵母细胞，研究了不同电场条件下脉冲次数和卵龄对卵母细胞活化效果的影响。     

中子活化法测定河水中的微量元素

本文阐述了用中子活化法分析新疆南部克里雅河河水中的微量元素,得到了河水中微量元素含量随河长及海拔高度而变化的分布规律,所得结果与克里雅河流域的不同地貌类型沉积相带基本相符,为分析河水的水化学特性开辟了新的途径。