QuEChERS-超高效液相色谱-串联质谱法同时测定大米、小麦和动物源食品中异丙隆及其代谢物脱甲基异丙隆残留

本研究建立了超高效液相色谱-串联质谱法(UHPLC-MS/MS)同时测定异丙隆及其代谢物脱甲基异丙隆在大米、小麦、牛肉、牛奶、鸡肉和鸡蛋的残留检测方法。样品经2%甲酸乙腈提取,以N-丙基乙二胺(PSA)净化,利用乙腈和0.2%甲酸水作为流动相梯度洗脱,T3色谱柱分离,在多反应监测模式下定量分析,基质外标法定量。结果表明:异丙隆及其代谢物脱甲基异丙隆溶剂标准曲线和基质标准曲线在1~1 000μg/L范围内线性关系良好,相关系数均大于0.99。在4个加标水平下,异丙隆日内平均回收率为74.0%~107.0%,相对标准偏差0.7%~12.9%;日间平均回收率为76.2%~108.7%,相对标准偏差1.1%~19.8%。脱甲基异丙隆日内平均回收率为76.9%~113.5%,相对标准偏差0.6%~13.9%;日间平均回收率为77.7%~107.4%,相对标准偏差2.2%~17.4%。异丙隆和脱甲基异丙隆的定量限均为1.0μg/kg。该方法简便、快捷、准确、灵敏度高,适用于异丙隆和脱甲基异丙隆在大米、小麦、牛肉、牛奶、鸡肉和鸡蛋6种基质中残留的检测,为解决异丙隆和脱甲基异丙隆在食品中残留的安全问题提供技术方法。     

Up-regulation of miR-181a attenuates releases of CSE-induced pro-inflammatory factors and expression of collagen IV,fibronectin and α-SMA in human bronchial epithelial cells

AIM: To investigate the effect and potential mechanism of microRNA-181a (miR-181a) on cigarette smoke extract (CSE)-induced the productions of pro-inflammatory factors and the expression of collagen IV, fibronectin and α-smooth muscle actin (α-SMA) in human bronchial epithelial cells (HBECs). METHODS: CSE-induced miR-181a expression was detected by RT-qPCR in the HBECs. After tansfected with miR-181a mimic, the releases of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6 and transforming growth factor-β1 (TGF-β1) were measured by ELISA, the protein expression of collagen IV, fibronectin and α-SMA was determined by Western blot. The activation of NF-κB/TGF-β1/Smad3 pathway was also evaluated by Western blot. RESULTS: CSE increased the levels of TNF-α, IL-1β, IL-6 and TGF-β1 and the expression of collagen IV, fibronectin and α-SMA, and decreased the expression of miR-181a in the HBECs (P<0.05). However, transfected with miR-181a mimic partially prevented the releases of TNF-α, IL-1β, IL-6 and TGF-β1, and inhibited the expression of collagen IV, fibronectin and α-SMA (P<0.05). Additionally, the activation of NF-κB/TGF-β1/Smad3 evoked by CSE was attenuated after transfected with miR-181a mimic. CONCLUSION: Up-regulation of miR-181a prevents the releases of CSE-induced pro-inflammatory factors and expression of collagen IV, fibronectin and α-SMA in the HBECs, and its mechanism may be related to the inhibition of NF-κB/TGF-β1/Smad3 pathway.     

SIRT1 promotes autophagy of pancreatic cancer cells induced by hypoxia via regulating FOXO1/RAB7 signaling pathway

AIM: To investigate the effect of SIRT1 on the autophagy of pancreatic cancer cells under hypoxia condition, and to analyze the underlying mechanism of regulating FOXO1/RAB7 signaling pathway. METHODS: Western blot and immunofluorescence methods were used to determine the expression of SIRT1 in the pancreatic cancer cells. The small interfering RNA targeting SIRT1 and SIRT1 over-expression plasmid were transfected into the pancreatic cancer Panc-1 cells. Confocal microscopy was used to detect the LC3 expression. Western blot was used to analyze the protein levels of LC3, p62 and FOXO1/RAB7 signaling pathway-related molecules. Co-immunoprecipitation was used to detected the protein interaction between SIRT1 and FOXO1. RESULTS: The expression level of SIRT1 in the nucleus of Panc-1 cells was increased under hypoxia condition. Compared with negative control under hypoxia condition, knock-down of SIRT1 expression attenuated the autophagy flux in the pancreatic cancer Panc-1 cells (P<0.05). Over-expression of SIRT1 increased the protein levels of FOXO1 and RAB7. On the contrary, knock-down of SIRT1 expression inhibited the protein levels of FOXO1 and RAB7. The protein interaction between SIRT1 and FOXO1 in the pancreatic cancer cells was observed. CONCLUSION: SIRT1 in pancreatic cancer Panc-1 cells under hypoxia condition is over-expressed in the nucleus. Down-regulation of SIRT1 inhibits autophagy and its mechanism may be related to FOXO1/RAB7 signaling pathway.     

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.     

番茄ShARPC5抗白粉病功能分析

【目的】白粉病（powdery mildew）是番茄生产上的重要病害,严重影响番茄的产量。番茄基因组测序工作的完成为抗病基因挖掘提供了重要的信息资源。ARP2/3（actin-related protein 2 and 3）复合体是肌动蛋白微丝骨架动力学的主要调控因子,能够参与包括响应外界胁迫等多种细胞学过程。本研究通过对番茄ARPC5（actin-related protein C5）进行克隆和抗病功能验证,为番茄基因组信息完善、抗病机制解析和分子育种等方面打下基础。【方法】从番茄LA1777（Solanum habrochaites）cDNA中PCR扩增ShARPC5,使用DNAMAN 6.0进行多序列比对;MEGA 6.0构建系统发育树;应用在线工具ProtComp v. 9.0进行亚细胞定位预测。利用实时荧光定量PCR（qRT-PCR）比较接种白粉菌（Oidium neolycopersici,On-Lz）后高感品种Moneymaker（MM）和高抗品种LA1777中番茄ARPC5的表达特征,分析白粉菌侵染与ARPC5表达的相关性。应用病毒诱导的基因沉默（virus-induced gene silencing,VIGS）技术进一步验证该基因在番茄中的抗病功能,观察沉默株和野生型株系接种后表型变化,利用台盼蓝和DAB染色法检测植株产生过敏性坏死和H₂O₂的能力,并检测ShARPC5沉默后一些与植物抗病相关标记基因的表达变化。采用农杆菌浸花法遗传转化拟南芥过表达ShARPC5植株,观察转基因和野生型株系接种后表型变化,并统计单病斑分生孢子数。【结果】从番茄品种LA1777中克隆到ShARPC5,编码132个氨基酸残基,包含一个保守的P16-Arc结构域。与番茄MM-白粉菌亲和互作相比,非亲和互作的番茄品种LA1777在接种白粉菌后,ShARPC5显著上调表达,尤其在接种后18 h。在番茄上沉默ShARPC5能够增加植株对白粉菌On-Lz的敏感性,防卫反应基因PR1b1显著下调表达。组织学观察显示与对照植株相比,ShARPC5沉默植株接种后诱导产生过敏性坏死和活性氧减少。在烟草上瞬时过表达ShARPC5能够诱导产生坏死斑。相反,在拟南芥上过表达ShARPC5能够增加植物的抗病性。【结论】ShARPC5是番茄响应白粉菌侵染的重要基因,可减轻番茄白粉病的发病程度,在番茄抗白粉病机理研究方面具有较大的应用价值,可作为番茄抗白粉病分子育种的一个候选基因。     

Genetic bases of source-,sink-,and yield-related traits revealed by genome-wide association study in Xian rice

The source-sink relationship determines the ultimate grain yield.We investigated the genetic basis of the relationship between source and sink and yield potential in rice.In two environments,we identified quantitative trait loci(QTL)associated with sink capacity(total spikelet number per panicle and thousand-grain weight),source leaf(flag leaf length,flag leaf width and flag leaf area),source-sink relationship(total spikelet number to flag leaf area ratio)and yield-related traits(filled grain number per panicle,panicle number per plant,grain yield per plant,biomass per plant,and harvest index)by genome-wide association analysis using 272 Xian(indica)accessions.The panel showed substantial variation for all traits in the two environments and revealed complex phenotypic correlations.A total of 70 QTL influencing the 11 traits were identified using 469,377 high-quality SNP markers.Five QTL were detected consistently in four chromosomal regions in both environments.Five QTL clusters simultaneously affected source,sink,source–sink relationship,and grain yield traits,probably explaining the genetic basis of significant correlations of grain yield with source and sink traits.We selected 24 candidate genes in the four consistent QTL regions by identifying linkage disequilibrium(LD)blocks associated with significant SNPs and performing haplotype analysis.The genes included one cloned gene(NOG1)and three newly identified QTL(qHI6,qTGW7,and qFLA8).These results provide a theoretical basis for high-yield rice breeding by increasing and balancing source–sink relationships using marker-assisted selection.     

秸秆还田对东北半干旱区玉米‖花生轮作系统土壤水热特性及作物产量的影响

以东北半干旱区玉米‖花生轮作系统为研究对象,试验设传统玉米单作、花生单作、不同秸秆还田量下(30%、60%和100%)玉米‖花生轮作5个处理。通过连续3年田间定位试验,研究玉米秸秆不同还田量对农田土壤水热特性、产量及土地当量比(LER)的影响。结果表明,与传统单作相比,秸秆还田处理玉米区播前0～40 cm土壤平均含水量提高了1.9%～3.9%,花生区提高了11.0%～13.9%;成熟期,秸秆还田花生区0:00～8:00平均地温比传统单作提高了0.2℃～1.1℃,10:00～22:00平均地温较传统单作降低了0.5℃～1.0℃。2016年和2017年玉米‖花生轮作系统LER为1.05～1.16,2018年各处理LER均小于1。吉林半干旱区玉米‖花生轮作系统进行秸秆还田改善了作物播前的土壤水分状况,但对于作物产量的影响在不同年份有所差异。     

小麦幼苗根系生理特性和解剖结构对根际pH的响应

为探讨根际pH 对冬小麦根系生长的影响，以半冬性小麦品种''矮抗58''（AK58）和''百农4199''（BN4199）为材料，采用水培试验，设置3个pH 水平（4.0、6.5、9.0，以6.5为对照），研究根际pH 对冬小麦根系抗氧化酶系统的影响及根系解剖结构对pH 的响应。结果表明：酸碱胁迫下，植株地上部及根系生物量、抗氧化酶活性较对照下降，且酸胁迫较碱胁迫下降幅度大。两品种根冠比表现为pH 4.0＜pH 6.5＜pH 9.0，丙二醛（MDA）含量表现为pH 6.5＜pH 9.0＜pH 4.0。与对照相比，两品种碱性条件下根系结构受损较小，而酸性条件下受损较大。相关分析表明地上部和根系干物质积累量与抗氧化酶活性呈显著正相关，与根系MDA含量呈极显著负相关。可见，酸碱胁迫下，保持较高的抗氧化酶活性有利于保持根系生长，从而提高小麦适应不良根际环境的能力。     

超级稻甬优12产量1000kg/667m~2手插栽培技术初步集成

以甬优12在居于5 300℃积温线的浙江中部新昌县的试验实践为基础,提出了1 000 kg/667 m2手插栽培的基本技术思路、群体生育指标及主要集成技术。