An improved method for extraction of sorghum polymeric protein complexes

A method for fractionating sorghum proteins using extraction solvents and techniques designed to obtain polymeric protein structures (especially disulfide linked) was developed. Extraction and separation conditions were optimized in terms of completeness of protein extraction, sample stability, and analytical resolution. After pre-extraction of albumins and globulins, a 3-step sequential procedure involving no reducing agents was applied to ground whole sorghum flour. The three fractions obtained represented proportionally different protein polymer contents and molecular weight distribution as evidenced by comparative size exclusion chromatography. Protein composition also varied among the extracts with differences in kafirin composition and non-kafirin proteins detected in the fractions by RP-HPLC and SDS-PAGE analysis. The ability to quantify and further characterize sorghum polymeric protein complexes will be useful for additional studies linking protein structures with functionality and digestibility and variations for these properties within diverse sorghum germplasm.     

秸秆还田与优化施氮改善烤烟根际土壤碳氮转化及氮素吸收

针对华南沙泥田烟区氮肥过量施用导致烟叶品质下降的问题,拟通过秸秆还田与氮肥优化配施调节烤烟各生育期氮吸收、根际土壤碳氮转化及相关酶的活性。试验采用随机区组设计,设水稻秸秆还田与氮肥两因素,其中3个碳（秸秆还田）水平：C0,无秸秆还田,0 kg/hm ²;C1,中量秸秆还田,4500 kg/hm ²;C2,全部秸秆还田,9000 kg/hm ²;2个氮水平：N1,传统施氮,169.50 kg/hm ²;N2,优化施氮,105.00 kg/hm ²。结果表明,与N1相比,N2处理秸秆还田下烤烟前期烟叶和根系氮吸收和累积正常,而后期有所下降;C1、C2处理时,与现蕾期相比,成熟期时根系氮含量均降低4.2 g/kg,叶片氮含量降低3.8、3.0 g/kg;成熟期时,与N1处理相比,N2处理时C1、C2处理的土壤无机氮含量降低30.4%、20.0%,前期2个氮处理间土壤无机氮含量无差异,优化施氮下中量秸秆还田（C1N2）处理烤烟土壤NH₄ ⁺-N含量在整个生育期内较高且呈降低趋势,而硝态氮和土壤无机氮含量在成熟期最低,表明中量秸秆还田下优化施氮能够维持生育前期土壤无机氮含量,且降低成熟期土壤无机氮含量。2个秸秆还田量处理时,团棵期N2处理的土壤转化酶活性是N1处理的1.30、1.13倍;旺长期土壤转化酶活性是N1处理的1.27、1.10倍;与传统施氮相比,秸秆还田下优化施氮处理增加了整个生育期土壤脲酶、磷酸酶、过氧化氢酶活性。在中量秸秆还田（C1）处理下,与N1水平相比,现蕾期和成熟期的N2处理土壤水溶性碳含量提高11.2%和14.1%;与N1相比,N2处理的土壤微生物量氮含量呈降低趋势。综上所述,优化施氮耦合中量秸秆还田（C1N2）能够提高土壤水溶性碳含量,维持土壤氮素合理供应,稳定烤烟生育前期对氮素吸收与累积,减少生育后期烟叶氮素奢侈吸收,与烤烟吸氮规律比较吻合。     

基于RNMU的多源星载SAR影像融合与土地覆盖分类

为充分利用多时相、多极化SAR数据在不同土地覆盖类型中的后向散射特性,将递归非负矩阵下近似(Recursive nonnegative matrix underapproximation,RNMU)算法引入多源SAR数据的融合,并利用融合后的SAR影像实现较高精度的土地覆盖分类。融合过程中,在根据不同模式SAR影像特点进行多源SAR影像预处理的基础上,基于RNMU算法通过对多个输入SAR影像进行矩阵分解及迭代最优矩阵求解,得到融合影像。为验证融合后SAR影像在土地覆盖分类中的应用效果,以吉林省大安市为研究区,对多时相Sentinel-1的VV/VH双极化SAR数据和高分三号(GF-3)的HH/HV双极化SAR数据进行了基于RNMU的影像融合,并利用融合后的SAR影像进行研究区主要土地覆盖类型分类。实验结果表明,基于RNMU融合影像的土地覆盖分类总体精度达93. 11%,Kappa系数为0. 86,与Gram-Schmid(G-S)融合方法相比,分类总体精度提高了6. 83个百分点,Kappa系数提高0. 12。多源SAR融合为SAR影像融合提供了有效手段,为土地覆盖分类提供了更多高精度的数据资源。     

ROCK promotes high glucose-induced cardiomyocyte apoptosis by inhi-biting PI3K/Akt signaling pathway

AIMTo investigate whether Rho-associated coiled-coil kinase (ROCK) is involved in high glucose-induced apoptosis of primary cardiomyocytes by regulating PI3K/Akt signaling pathway. METHODSPrimary Wistar rat cardiomyocytes were cultured and identified by α-sarcomeric actin (α-SCA) immunohistochemistry. Cardiomyocytes were treated with 5.5, 33 and 40 mmol/L glucose for 48 h. The cell viability was measured by MTT assay, and the mRNA expression of ROCK1 and ROCK2 in the cardiomyocytes was detected by RT-qPCR. Flow cytometry was used to analyze the apoptosis of the cardiomyocytes. The protein levels of ROCK1, ROCK2, cleaved caspase-3, Bcl-2, PI3K, Akt and p-Akt were determined by Western blot. In order to confirm the regulatory effect of ROCKs on PI3K/Akt signaling pathway, the cells were divided into control group (5.5 mmol/L glucose), high glucose group (33 mmol/L glucose) and high glucose+Y27632 (ROCK inhibitor) group. Western blot was used to detect the protein levels of ROCK1, ROCK2, PI3K, Akt and p-Akt. RESULTSAfter 48 h of high glucose exposure, the values of relative cell viability in 33 and 40 mmol/L glucose groups were (79.71±2.43)% and (68.41±7.49)%, respectively, both of which were significantly decreased compared with normal control group (P<0.05). After 48 h of high glucose exposure, the relative mRNA levels of ROCK1 and ROCK2 in 33 and 40 mmol/L glucose groups were significantly increased compared with normal control group (P<0.05). Compared with normal control group, the apoptotic rate in 33 and 40 mmol/L glucose groups was increased significantly (P<0.05). Compared with normal control group, the protein expression of ROCK1, ROCK2 and cleaved caspase-3 in 33 and 40 mmol/L glucose groups was increased (P<0.05), while the protein expression of Bcl-2 was decreased (P<0.05). No significant difference in the protein levels of PI3K and Akt among the 3 groups was observed, while the protein level of p-Akt in 33 and 40 mmol/L glucose groups was decreased compared with normal control group (P<0.05). Compared with high glucose group, the expression of ROCK1 and ROCK2 was decreased in high glucose+Y27632 group. No significant difference in the protein levels of PI3K and Akt among the 3 groups was observed. Compared with normal control group, the protein level of p-Akt in high glucose group was decreased, and the protein level of p-Akt in high glucose+Y27632 group was increased significantly compared with high glucose group. CONCLUSION Under high glucose environment, ROCK may reduce the level of p-Akt by inhibiting the PI3K/Akt signaling pathway， thus promoting the apoptosis of cardiomyocytes.     

芜菁Br14-3-3的克隆及其在非生物胁迫下的表达分析

克隆了‘津田芜菁’（Brassica rapa subsp. rapifera‘Tsuda’）通用调节因子14-3-3基因cDNA序列，命名为Br14-3-3（GenBank登录号为MK896872）。该基因全长1 113 bp，开放阅读框全长774 bp，编码含有257个氨基酸的多肽。荧光定量PCR分析Br14-3-3在‘津田芜菁’不同组织中及其在温度、脱水、渗透、ABA和无机盐等非生物胁迫下幼苗中的表达，结果表明该基因在‘津田芜菁’的花中表达量最高，幼苗中次之；低温抑制了Br14-3-3的表达，其他非生物胁迫可诱导该基因表达，暗示Br14-3-3在非生物胁迫应答中发挥功能。     

赤霉素不同处理浓度和浸种时间对托鲁巴姆发芽的影响

托鲁巴姆种子的休眠较深,发芽较困难。为了打破其种子的休眠期,加快种子萌发,提高种子发芽的整齐度,以托鲁巴姆为试材,研究了不同的赤霉素处理浓度和浸种时间对其发芽的影响。结果表明:生产中适宜的赤霉素处理浓度为750~1 250 mg/L,浸种时间为36 h,发芽率可达93.00%;可有效提高托鲁巴姆种子的发芽速率和发芽整齐度。     

Temperature modulates liver lipid accumulation in Atlantic salmon (Salmo salar L.) fed low dietary levels of long‐chain n‐3 fatty acids

Atlantic salmon (Salmo salar) were fed five graded levels of eicosapentaenoic acid (EPA, 20:5n‐3) and docosahexaenoic acid (DHA, 22:6n‐3), from 1.4 to 5.2% of total fatty acids (FA, 5–17 mg kg^?1 feed), and grew from ~160 g to ~3000 g, with the period from 1450 g onwards conducted both at 6 °C and at 12 °C. All fish appeared healthy, and there were no diet‐related differences in haematological or plasma parameters, as well as intestinal histological or gut microbiota analysis. Fish reared at 6 °C had higher accumulation of storage lipids in the liver compared to fish reared at 12 °C. Liver lipids also increased with decreasing dietary EPA + DHA at 6 °C, while there was no such relationship at 12 °C. Gene expression of SREBP1 and 2, LXR, FAS and CPT1 could not explain the differences in liver lipid accumulation. In liver polar lipids, DHA was found to be reduced when dietary EPA + DHA was <2.7% of FAs, while the level of EPA in the membranes was not affected. In conclusion, reducing dietary EPA + DHA from 5.2 to 1.4% of total FAs had a minor impact on fish health. Temperature was the factor that most affected the liver lipid accumulation, but there was also an interaction with dietary components.     

圆柱镂空型人工鱼礁波流水动力特性数值模拟

研究人工鱼礁在波流作用下的水动力特性,对于人工鱼礁的设计具有重要的意义。基于有限体积法,采用边界造波,利用自由表面捕捉法(VOF)捕捉自由水面,建立了可以分别模拟纯波、均匀流以及波流共同作用下人工鱼礁水动力特性的多功能三维数值波流水槽。基于该数值模型对不同波流工况作用下圆柱型镂空人工鱼礁水动力特性进行数值模拟,并与物理模型试验结果进行比较。结果显示,人工鱼礁数值模拟受力与模型试验结果吻合良好,人工鱼礁所受的波流力最大值随着波高、周期和水流流速的增大而增大;人工鱼礁处于波流场波峰正下方时,背涡流的面积随着水流流速的增大而增大,随着波高、周期增大而减小。对单独均匀流作用、单独波浪作用和波流联合作用下人工鱼礁的水动力特性对比研究表明,人工鱼礁所受的最大波流力比最大波浪力、水流力都大,波流联合作用下的流场效应最显著,在礁体的后部形成了较大规模的漩涡结构。     

Dietary α‐linolenic acid affects lipid metabolism and tissue fatty acid profile and induces apoptosis in intraperitoneal adipose tissue of juvenile grass carp (Ctenopharyngodon idella)

A 56‐day feeding trial was conducted to elucidate the effects and mechanism action of dietary α‐linolenic acid (ALA, 18:3n‐3) on lipid accumulation and fatty acid profile of muscle, hepatopancreas and intraperitoneal fat (IPF) in juvenile grass carp using three isonitrogenous and isoenergetic semi‐purified diets containing 0.0% (control group), 1.0% and 2.0% ALA, respectively. The lowest intraperitoneal fat (IPF) ratio was found in 2.0% group. In the muscle, hepatopancreas and IPF, docosahexaenoic acid (DHA, 22:6n‐3) and eicosapentaenoic acid (EPA, 20:5n‐3) contents increased with the increase in dietary ALA. In the IPF, caspase 3, caspase 8 and caspase 9 showed the highest activities in 2.0% group, while the value of Bcl‐2/Bax (B‐cell leukaemia 2/Bcl‐2‐associated X protein) reached the lowest. Meanwhile, swelling of the IPF mitochondria was observed in 2.0% group. The gene expressions of fatty acid desaturase (FAD) and fatty acid elongase (ELO) in the hepatopancreas and muscle showed significantly higher levels in the treatment groups, whereas an opposite trend was existed in the IPF. Fatty acid synthase (FAS), sterol regulatory element binding protein‐1c (SREBP‐1c) in the IPF and hepatopancreas reached the lowest in 2.0% group. Overall, dietary ALA could promote n‐3 highly unsaturated fatty acids (HUFAs) synthesis and suppress the accumulation of lipid by decreasing the expression of related genes and promoting the apoptosis in IPF.     

茶树GGPS基因家族的克隆、生物信息学和表达分析

牻牛儿基牻牛儿基焦磷酸合酶(Geranylgeranyl diphosphate synthase,GGPS)是萜类合成途径的结构酶,对植物生长发育具有重要意义。本研究通过RACE和RT-PCR方法克隆得到5条潜在的茶树GGPS序列,分别命名为CsGGPS1-4和CsGGPS9,其中CsGGPS9存在3条等位基因,分别是CsGGPS9-1、CsGGPS9-2和CsGGPS9-3,在系统进化树上与其他基因分成两支。蛋白质序列分析表明,茶树GGPS家族成员都具有polyprenyl_synt结构域,不存在信号肽序列。亚细胞定位预测结果显示,CsGGPS1、CsGGPS2和CsGGPS4定位在叶绿体上,CsGGPS3和CsGGPS9定位在线粒体上。通过Swiss Model进行三维建模,结合"three-floor"模型对茶树GGPS家族成员的功能进行预测,预测结果显示,CsGGPS1、CsGGPS2和CsGGPS4是GGPS;CsGGPS3是异源二聚体形式的牻牛儿基焦磷酸合酶的小亚基;CsGGPS9的催化主产物是碳链数大于30的异戊烯基焦磷酸。q RT-PCR分析表明,CsGGPS1整体表达丰度较低,仅在一芽二叶中表达量稍高;CsGGPS2在茶树各个组织中均有表达,在花中表达量最高,且花发育过程中表达量先上升后下降;CsGGPS3在叶和幼根中的表达量高于花,花发育过程中表达平稳;CsGGPS4在茶树各个组织中表达量数值相近,在花发育过程中表达量变化趋势与CsGGPS2相同;CsGGPS9的表达量在成熟叶中显著低于幼嫩叶片。