芒果中性/碱性转化酶MiNI基因的克隆与表达分析

采用RT-PCR技术克隆了1个编码NI基因的cDNA序列,命名为MiNI基因,并对不同来源的中性/碱性转化酶的分子特征及系统进化进行比较分析。结果表明：MiNI基因开放阅读框为2034 bp,编码677个氨基酸,相对分子量为76.6 kDa,理论等电点为6.24;生物信息学分析结果显示,NI二级结构α螺旋占38.85%,无规则卷曲占35.45%,伸展链占18.91%,β折叠占6.79%;MiNI具有glycoside hydrolase family 100结构保守域,与克里曼丁桔、龙眼、巴西橡胶树和番木瓜都具有一致的motif位点;MiNI基因编码的氨基酸序列与克里曼丁桔、龙眼氨基酸序列同源性最高;构建NI系统进化树分析表明,与芒果遗传距离最近的是克里曼丁桔,最远的是玉米和枸杞。qRT-PCR分析显示,果皮MiNI基因表达量远高于果肉;花后10~40 d,果皮MiNI基因表达量显著下降;花后40~100 d,果皮MiNI基因表达量维持在一个相对稳定水平;花后100~130 d,随着果实成熟,果皮MiNI基因表达量又显著上升;而花后10~40 d,果肉MiNI基因表达量显著下降,至果实发育后期果肉MiNI基因表达量始终处于极低水平。该研究为进一步了解MiNI基因在芒果果实蔗糖代谢过程中的作用以及从分子角度阐明芒果糖代谢机理奠定理论和技术基础。     

Basal internode elongation of rice as affected by light intensity and leaf area

Short basal internodes are important for lodging resistance of rice(Oryza sativa L.).Several canopy indices affect the elongation of basal internodes,but uncertainty as to the key factors determining elongation of basal internodes persists.The objectives of this study were(1)to identify key factors affecting the elongation of basal internodes and(2)to establish a quantitative relationship between basal internode length and canopy indices.An inbred rice cultivar,Yinjingruanzhan,was grown in two split-plot field experiments with three N rates(0,75,and 150 kg N ha−1 in early season and 0,90,and 180 kg N ha−1 in late season)as main plots,three seedling densities(16.7,75.0,and 187.5 seedlings m−2)as subplots,and three replications in the 2015 early and late seasons in Guangzhou,China.Light intensity at base of canopy(Lb),light quality as determined from red/far-red light ratio(R/FR),light transmission ratio(LTR),leaf area index(LAI),leaf N concentration(NLV)and final length of second internode(counted from soil surface upward)(FIL)were recorded.Higher N rate and seedling density resulted in significantly longer FIL.FIL was negatively correlated with Lb,LTR,and R/FR(P<0.01)and positively correlated with LAI(P<0.01),but not correlated with NLV(P>0.05).Stepwise linear regression analysis showed that FIL was strongly associated with Lb and LAI(R2=0.82).Heavy N application to pot-grown rice at the beginning of first internode elongation did not change FIL.We conclude that FIL is determined mainly by Lb and LAI at jointing stage.NLV has no direct effect on the elongation of basal internodes.N application indirectly affects FIL by changing LAI and light conditions in the rice canopy.Reducing LAI and improving canopy light transmission at jointing stage can shorten the basal internodes and increase the lodging resistance of rice.     

Change features of time-series climate variables from 1962 to 2016 in Inner Mongolia,China

Detecting change features of climate variables in arid/semi-arid areas is essential for understanding related climate change patterns and the driving and evolution mechanism between climate and arid/semi-arid ecosystems.This paper takes Inner Mongolia of China,a unique arid/semi-arid ecosystem,as the study area.We first detected trend features of climate variables using the linear trend analysis method and then detected their trend-shift features using the breaks for additive seasonal and trend method based on the time-series of monthly precipitation and monthly mean temperature datasets from 1962 to 2016.We analyzed the different change features of precipitation and temperature on a regional scale and in different ecological zones to discover the spatial heterogeneity of change features.The results showed that Inner Mongolia has become warmer-wetter during the past 54 years.The regional annual mean temperature increased 0.4°C per decade with a change rate of 56.2%.The regional annual precipitation increased 0.07 mm per decade with a slightly change rate of about 1.7%,but the trend was not statistically significant.The warmer trend was contributed by the same positive trend in each season,while the wetter trend was contributed by the negative trend of the summer precipitation and the positive trend of the other three seasons.The regional monthly precipitation series had a trend-shift pattern with a structural breakpoint in the year 1999,while the regional monthly mean temperature series showed an increasing trend without a periodical trend-shift.After the year 2000,the warmer-wetter trend of the climate in Inner Mongolia was accelerated.The late 20th century was a key period,because the acceleration of the wetter trend in some local zones(I and II)and the alleviation of the warmer trend in some local zones(Ⅶ,Ⅷand IX)occurred simultaneously.Moreover,the change features had a strong spatial heterogeneity,the southeastern and southwestern of Inner Mongolia went through a warmer-drier trend compared with the other areas.The spatio-temporal heterogeneity of the climate change features is a necessary background for various types of research,such as regional climate change,the evolution of arid/semi-arid ecosystems,and the interaction mechanisms between climate and arid/semi-arid ecosystems based on earth-system models in Inner Mongolia.     

Hyperbaric oxygen therapy activates CREB/BDNF signaling pathway to reduce synaptic damage in hippocampal neurons of Alzheimer disease mice

AIM To investigate the effect of hyperbaric oxygen (HBO) on synaptic damage of hippocampal neurons in APP/PS1 transgenic (TG) mice and its possible mechanism. METHODS The 6-month-old male APP/PS1 TG mice were randomly divided into TG group, HBO group and cAMP response element binding protein (CREB) inhibitor H89 group, with 10 mice in each group. Ten male wild-type (WT) C57BL/6 mice of the same age were used as negative control group (WT group). The mice in HBO and H89 groups were treated with HBO for 6 cycles, while the mice in WT group and TG group were not treated. The learning and memory abilities were observed by Morris water maze. The nesting ability of the mice was detected by nesting test. The Nissl bodies in hippocampal neurons were observed by Nissl staining. The mRNA expression of CREB and brain-derived neurotrophic factor (BDNF) in hippocampus was detected by real-time PCR. The protein levels of synapsin (SYN), postsynaptic density protein 95 (PSD95), growth-associated protein 43 (GAP43), CREB, phosphorylated CREB (p-CREB) and BDNF in the hippocampus were determined by Western blot. RESULTS Compared with WT group, the learning and memory abilities of the mice in TG group were signilficantly reduced (P<0.05). In addition, the nesting score, the number of Nissl bodies in the hippocampal neurons, the mRNA expression of CREB and BDNF, and the protein levels of SYN, PSD95, GAP43, p-CREB and BDNF were also decreased significantly (P<0.05). Compared with TG group, the learning and memory abilities of the mice in HBO group were improved (P<0.05). Meanwhile, the nesting scores of the mice were significantly increased (P<0.05), the neurons in the hippocampus were arranged neatly, and the number of Nissl bodies, the relative mRNA expression of CREB and BDNF,and the protein levels of SYN, PSD95, GAP43, p-CREB and BDNF were also increased significantly (P<0.05). Compared with HBO group, the mice in H89 group had poor learning and memory abilities, lowered nesting scores and decreased number of Nissl bodies. Futhermore, the relative mRNA expression of CREB and BDNF, and the protein levels of SYN, PSD95, GAP43, p-CREB and BDNF were also decreased significantly (P<0.05). CONCLUSION HBO improves the learning and memory abilities of APP/PS1 TG mice, and its mechanism may be related to activating the CREB/BDNF signaling pathway to reduce synaptic damage of hippocampal neurons in mice.     

Generation of a uniform thymic malignant lymphoma model with C57BL/6J p53 gene deficient mice

Lymphoma is the third most common cancer diagnosed in children, and T-cell lymphoma has the worst prognosis based on clinical observations. To date, a lymphoma model with uniform penetrance has not yet been developed. In this study, we generated a p53 deficient mouse model by targeting embryonic stem cells derived from a C57BL/6J mouse strain. Homozygous p53 deficient mice exhibited a higher rate of spontaneous tumorigenesis, with a high spontaneous occurrence rate (93.3%) of malignant lymphoma. Because tumor models with high phenotypic consistency are currently needed, we generated a lymphoma model by a single intraperitoneal injection of 37.5 or 75 mg/kg N-methyl-N-nitrosourea to p53 deficient mice. Lymphoma and retinal degeneration occurred in 100% of p53^+/− mice administered with higher concentrations of N-methyl-N-nitrosourea, a much greater response than those of previously reported models. The main anatomic sites of lymphoma were the thymus, spleen, bone marrow, and lymph nodes. Both induced and spontaneous lymphomas in the thymus and spleen stained positive for CD3 antigen, and flow cytometry detected positive CD4 and/or CD8 cells. Based on our observations and previous data, we hypothesize that mice with a B6 background are prone to lymphomagenesis.     

壳聚糖/海藻酸钠微球对红景天苷控制释放的研究

将含有药物的海藻酸钠溶液滴入到壳聚糖和氯化钙的混合溶液中形成微球,制备一系列红景天苷微球,研究微球对红景天苷的包载能力及释药特性。结果表明:海藻酸钠、氯化钙、壳聚糖的质量浓度、海藻酸钠与红景天苷的比例及壳聚糖溶液pH值对微球的包埋率、载药率及缓释性能有影响,而成膜反应时间对载药率和包埋率有影响,对缓释性能没有影响。缓释效果最佳的微球制备工艺条件为海藻酸钠与红景天苷质量比为1.5,海藻酸钠2.5 g/mL,壳聚糖0.8 g/mL,氯化钙1.5 g/mL,成膜时间为5 min,pH值为5.5。     

水文分异条件下光养生物膜藻类对氮/磷的响应

光养生物膜是河流生态系统中重要的初级生产者,在河流生物地球化学循环中扮演重要角色。然而,河流水文状态及营养盐差异引起的生境异质性对光养生物膜藻类的影响未知。本研究在微型跑道池模拟流水(0.5 m/s)和静水(0 m/s)条件的基础上,通过设置不同浓度氮(1.51, 2.51和5.51 mg/L)、磷(0.1, 0.2和0.4 mg/L)及氮磷比(8, 16和32)条件培养野外采集的原位生物膜及人工基质,探究水文分异和营养变化对河流光养生物膜藻类物种组成及密度的影响。非度量多维排序(NMDS)分析结果表明,差异水文条件下,原位和建群生物膜藻类群落在梯度氮、磷和氮磷比环境中均呈现明显分离(PERMANOVA,P < 0.001),且建群生物膜中各分组藻类群落具有更明显的差异。双因素方差分析结果表明,生物膜中硅藻、蓝藻和绿藻的生长对营养盐与水文变化的响应并不一致,其中磷处理中,磷与流速单一及其交互效应显著影响了大多数藻类的生长及建群(P < 0.001);而氮处理中,氮与流速的交互效应仅对建群生物膜藻类影响显著(P < 0.001)。研究结果也发现,藻类在静水环境更有利于建群生物膜的形成,且静水-高营养盐环境更有利于蓝藻和绿藻的生长。这些结果说明,生物膜建群初期易受到水文扰动的影响,且水文分异和氮、磷影响了光养生物膜藻类的响应模式,研究为河流生态保护提供了新视角。     

5种动物源食品中的氟虫腈及其3个代谢物残留量检测

利用QuEChERS技术结合超高效液相色谱-串联质谱的方法检测5种动物源食品(鸡肉、鸡蛋、猪肝、牛奶、猪肉)中氟虫腈及其3个代谢物残留量。样品用乙腈提取,经低温处理,多壁碳纳米管(MWCNTs)、弗罗里硅土(Florisil)和C18分散固相萃取(d-SPE)净化,外标法定量。在不同浓度的加标水平下,氟虫腈及其代谢物在5种动物源食品中的平均回收率为72.1%～113.8%,相对标准偏差RSD为1.9%～17.6%,检出限LOD在0.73～1.94μg/kg范围内,定量限LOQ在5～10μg/kg范围内。该方法具有简单、灵敏、准确等优点,适用于动物源食品中氟虫腈及其代谢物的快速筛查和定量检测。     

Structural and Rheological Characteristics of Alginate from Sargassum cristaefolium Extracted by Twin Screw Extruder

This study aimed to investigate the effects of process extrusion on the characteristics of Sargassum cristaefolium sodium alginate (SCSA) extracted using twin-screw extruder. Box-Behnken Design (BBD) from response surface methodology (RSM) was established to understand the effects of temperature, screw speed, and pH on the multiple-responses of alginate characteristics including intrinsic viscosity, yield, and molecular weight. The results revealed that temperature, screw speed, and pH significantly affected (P < .05) all responses. The optimum extraction condition was found at temperature of 58.18°C, screw speed of 77.99 rpm, and pH 10.11. At this condition, the response of residence time distribution was 7.07 ± 0.029 min, yield of 34.01 ± 0.12%, intrinsic viscosity of 460.13 ± 14.75 mL/g, and molecular weight of 217.94 ± 7.14 × 10³ g/mol. This alginate had mannuronic acid to guluronic acid (M/G) ratio of 0.28, and the L-guluronic acid block was 0.78, which was higher than the D-mannuronic acid block. Rheological characterization of SCSA in aqueous solution was shear-thinning pseudoplastic, and alginate gel in 1 M CaCl₂ was more elastic than liquid.