加味大柴胡汤对阻塞性黄疸老年大鼠TRPC1表达的影响

目的 探讨加味大柴胡汤在临床上辅助治疗老年阻塞性黄疸的理论机制。方法 通过结扎胆总管的外科手术方法建立阻塞性黄疸老年大鼠模型,实验分为假手术组、模型组、加味大柴胡汤组,予以相应干预,分3、7、10 d共3个时间段,每组10只SPF级老年SD大鼠,在各时间段取老年大鼠肝组织及心脏血,检测AST、ALT及TBA;采用钙离子试剂盒检测老年鼠肝组织匀浆钙离子浓度;采用Western Blot法检测老年鼠肝组织TRPC1蛋白的表达,采用Tunel法观察肝细胞凋亡情况。结果 造模成功后各时间点模型组、加味大柴胡汤组与假手术组比较,ALT、AST、TBA和Ca²⁺浓度均升高,且模型组上升更明显（P<0.01）,模型组、加味大柴胡汤组的TRPC1的蛋白含量也均高于假手术组（P<0.01）,且加味大柴胡汤组肝细胞的凋亡率低于模型组（P<0.01）。结论 梗阻性黄疸老年大鼠TRPC1表达明显升高,加味大柴胡汤下调TRPC1的表达,可能是其降低钙超载,减少肝细胞凋亡的机制之一。     

Phenotypic Analysis and Gene Cloning of Rice Floury Endosperm Mutant fse4

【Objective】Starch is the main energy reserve of rice endosperm. The biosynthesis of starch is complex, requiring a large number of synthetic enzymes and regulators. Screening rice endosperm defective mutants and cloning the underlying genes will lay theoretical basis for starch biosynthesis and its regulation. 【Method】 A stable genetic floury and shrunken endosperm mutant termed as fse4 (floury and shrunken4) were obtained from the mutant library of Ningjing 3 (WT), which was induced by N-methyl-N-nitrosourea (MNU). An F2 mapping population was generated by crossing the fse4 mutant with Dular (an indica rice variety) and the gene was finally isolated. The floury seeds segregated from the fse4 heterozygous plants were used to observe the morphological features, and the physicochemical properties of the brown rice flour were analyzed. The endosperm structure was observed with a scanning electron microscopy by the semi-thin section technology. The expression of starch synthesis related genes during grain filling was determined by qRT-PCR; Immunoblotting was used to detect the accumulation of proteins related to starch synthesis. The amino acids contents of each mature endosperm were determined with the fully automatic amino acid analyzer.【Result】The 1000-grain weight and grain size were significantly reduced in fse4. Compared with WT, the contents of total starch, amylose and total protein were signi?cantly lower in fse4, while the lipid content was signi?cantly higher. The starch viscosity, breakdown viscosity and setback viscosity of the fse4 mutant were lower than WT. The endosperm of the mutant had many single dispersed starch granules with large spaces between each other. Using 1568 recessive individuals, FSE4 was narrowed down to a 252 kb region. Sequencing revealed a single base substitution in the first exon of the delta 1-pyrroline-5-carboxylate synthetase (P5CS), resulting in a conserved amino acid variation. Most of the genes related to starch synthesis were downregulated in fse4 and the protein accumulation related to starch synthase were reduced. The contents of various amino acids in fse4 rice flour were increased or decreased, the total free amino acids contents in fse4 seeds was 2.6 times higher than those in WT. Exogenous proline was applied during the germination of fse4 seeds, and the embryonic lethal phenotype was partially recovered.【Conclusion】FSE4 encode the key rate-limiting enzyme P5CS of proline synthesis, which plays an important role in the biosynthesis and metabolism of amino acids in endosperm and affects the accumulation of starch.     

特色稻桂红糯1号品种特性及机械化栽培技术

发展糯稻特色产业对提高稻农效益具有重要意义。桂红糯1号是广西农业科学院水稻研究所选育的籼型常规红米糯稻品种,不同播种量、机插密度和施氮量对其产量及穗粒结构的影响试验表明,桂红糯1号机械化栽培需要适当增加播种量,提高机插密度,合理控制氮肥用量。本文介绍了桂红糯1号的特征特性及机械化栽培技术。     

花生脂肪酸延长酶基因ＡｈＦＡＥ１及其启动子的克隆与功能分析

为探讨花生脂肪酸中低芥酸含量的原因,从花生中克隆了脂肪酸延长酶FAE1及其启动子序列,并进行了功能分析。结果表明,AhFAE1全长cDNA为2 202bp,含有441bp的5?非翻译区及201bp的3?非翻译区,编码蛋白含519aa,分子量58.17Da,理论等电点9.15,AhFAE1与麻疯树（Jatropha curcas ALB76796.1）、黄麻（Corchorus capsularis OMO87584.1）、拟南芥AtKCS4亲缘关系较近。亚细胞定位结果显示AhFAE1定位于内质网。qRT-PCR结果显示,AhFAE1在各个组织中均有表达,在种子中随着种子的发育成“钟”型变化,花后60d表达量最高。构建了植物表达载体,通过农杆菌介导法转化拟南芥研究启动子功能,利用GUS组织化学染色研究其表达特征,在任何组织中未发现GUS活性。推测AhFAE1可能参与了花生长链脂肪酸的合成,但是该基因启动子转录活性弱可能是造成花生中低芥酸含量的主要原因。      

白粉菌诱导下小麦TaNHO1基因的克隆、表达分析及亚细胞定位

非寄主抗性基因 NHO1(non-host resistance 1)编码的甘油激酶(glycerol kinase,GK),是甘油代谢中的限速酶之一,参与植物对多种病害的抵御过程。为进一步探究其响应小麦白粉菌侵染的表达模式,利用RT-PCR方法克隆获得了转录自抗病种质N9134的三个部分同源染色体的小麦 NHO1基因(分别命名为 TaNHO1-2A、 TaNHO1-2B和 TaNHO1-2D),分析三个 TaNHO1同源基因的序列、启动子组成元件和表达模式,并明确了其亚细胞定位。序列分析结果表明,小麦 TaNHO1-2A/2B/2D基因CDS区序列全长分别为1 605、1 599和1 605 bp,分别编码534、532和534个氨基酸残基;3个同源基因均含有与 AtNHO1(AT1G80460.1)基因以及 OsNHO1(Os04G0647800)基因高度相似的FGGY_N端和FGGY_C端结构域。经对启动子区结构分析,该基因启动子区含有大量与植物激素及逆境响应相关的顺式作用元件,意味着 TaNHO1可受到多种植物激素诱导并参与小麦抗病过程。qRT-PCR结果表明,在N9134抗病近等基因系响应白粉菌(Blumeria graminis f. sp.tritici,Bgt)侵染过程中,三个同源基因在不同时间点表达模式不同,其中 TaNHO1-2A、 TaNHO1-2B基因在侵染早期均上调表达,而 TaNHO1-2D则表现出多次波动的表达模式;在N9134感病近等基因系遗传背景下,同源基因的表达水平在病菌侵染后48 h均表现出下调,说明该基因能够响应白粉菌侵染。经亚细胞定位分析,该基因主要作用于细胞质、细胞膜以及核膜。     

新疆巴什拜羊不同毛色与有关基因多态性分析

【目的】以不同毛色(红棕色73只,黑色53只,青灰色40只)新疆巴什拜羊为研究对象,采用PCR-SSCP技术检测ASIP、TYR、TYRP1和TYRP2基因多态性,并分析其与被毛表型的相关性。【方法】采用随机抽样的方法选取出三种毛色的巴什拜羊,用枸橼酸钠抗凝管进行采血。运用软件PIC-CALC和POPGENE 32计算巴什拜羊ASIP、TYR、TYRP1和TYRP2基因的多态信息含量和基因型频率、等位基因频率、纯合度、杂合度、有效等位基因数,并对该位点基因型的分布进行Hardy-Weinberg平衡的卡方检验。将所得数据输入Excel表中进行整理,并用SPSS 19.0软件进行差异性分析。【结果】ASIP基因在不同毛色的巴什拜羊群体中有两种基因型分别为AA型和AB型,等位基因B为红色被毛群体中的优势等位基因;等位基因A为黑色和青灰色两种被毛群体中的优势等位基因。处于Hardy-Weinberg平衡状态(P＞0.05),红色被毛多态信息含量处于中度多态,而黑色和青灰色被毛多态信息含量处于低度多态。TYR基因在不同毛色的巴什拜羊群体中有两种基因型分别为AA型、AG型和GG型,等位基因G为3种毛色的优势等位基因TYR基因在巴什拜羊黑色和青色被毛群体中处于Hardy-Weinberg平衡状态(P＞0.05),而红色被毛群体中处于不平衡状态(P<0.05)。TYRP1基因存在两种基因型:AA型和GG型,等位基因A为红色和青灰色两种毛色的优势等位基因;等位基因G为黑色被毛群体的优势等位基因。TYRP1基因在巴什拜羊群体中处于Hardy-Weinberg平衡状态。红色被毛多态信息含量处于高度多态,黑色和青灰色被毛多态信息含量处于低度多态。TYRP2基因PCR-SSCP检测结果没有显示多态。【结论】ASIP、TYR、TYRP1和TYRP2基因多态性与巴什拜羊的毛色性状有相关性。     

基于GF-1影像NDVI年度间相关分析的冬小麦面积变化监测

为实现区域冬小麦种植面积变化的快速监测,减少监测难度,提高监测效率和精度,该文提出一种基于年际NDVI相关关系的监测方法(relationship analysis of normal difference vegetation index,rNDVI)。选择河北省黄骅市、孟村县、海兴县3个县市为研究区,基于2014年4月14日、2017年4月26日两个时期的GF-1/WFV数据,基于rNDVI方法,通过将样本点两年度的NDVI值构建二维空间,采用最小二乘法拟合的方法获得不变地物点的上下包络线方程,进而得到冬小麦变化区域的监测阈值,提取冬小麦种植增加和减少区域,实现对研究区域的变化监测。结果表明,采用rNDVI算法总体精度分别为90.60%,Kappa系数为0.84,相比传统的先最大似然分类后再提取冬小麦种植变化区域的方法,总体精度与Kappa系数分别提高了6.6个百分点和16.7%。对冬小麦增加区域、冬小麦减少区域的变化监测结果进行分析,发现基于rNDVI的变化监测方法可以有效提高裸地、线状道路、破碎的冬小麦地块等区域的变化识别能力,提高监测精度。同时分别利用2014年3月1日和2017年3月12日、2014年5月17日与2017年5月20日两对GF-1/WFV数据进行基于rNDVI的冬小麦变化区域监测,结果表明3月份的监测精度较低,主要是由于3月份冬小麦长势尚不明显,5月份与4月份的总体精度相近,主要是由于5月份冬小麦NDVI已较高,易于识别。上述研究结果表明,基于rNDVI的冬小麦变化快速监测方法可以有效监测区域冬小麦种植面积的变化情况,算法简单高效,且能够在种植结构相对单一的冬小麦分布区域保持较高精度,能够满足农情遥感监测信息快速获取的需要。     

基于关联规则面向对象的海岸带海水养殖模式遥感识别

针对目前海水养殖模式遥感识别中的效率低,"同物异谱"、"异物同谱"和"椒盐"噪声等问题,该文研究了关联规则分类和面向对象相结合的养殖模式遥感识别方法,通过不同养殖模式的对象分割和关联规则的自动和智能获取,来构建海水养殖模式分类器。以高分一号PMS1卫星影像为数据源,把不同养殖模式对象的光谱、空间形态和纹理特征及其关联关系作为事务数据,使用Apriori算法挖掘类别作为后件的强规则,对粤东柘林湾养殖核心区内4种海水养殖模式(池塘养殖、网箱养殖、滩涂插养、浮筏吊养)水面信息进行提取。结果表明:基于关联规则面向对象的海水养殖模式分类精度能达到88.65%,比K-近邻法面向对象法精度提高了14.38个百分点,比关联规则挖掘分类法精度提高了12.16个百分点。关联规则分类和面向对象结合方法拓宽了传统逻辑推理分类方法中获取信息的途径,使分类更加自动化和智能化,且分类精度得到显著提高,可以成为海岸带海水养殖复杂模式识别的有效支持手段。     

PRRSV 2b蛋白与LGALS1相互作用的验证研究

旨在研究猪繁殖与呼吸综合症病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)2b蛋白与宿主细胞蛋白之间的相互作用。本研究利用酵母回返试验、GST-pull down试验、免疫共沉淀试验3种方法对LGALS1与PRRSV 2b蛋白间是否相互作用进行验证。结果表明:在酵母回返试验中,含有LGALS1与2b基因的杂交酵母菌株在SD/-Trp-Leu-His-Ade/AbA/X-α-gal的平板上生长出蓝色菌落;在GST-pull down试验中,LGALS1与2b蛋白反应的样品经Western blot可检测到融合蛋白带;在免疫共沉淀试验中,LGALS1与2b蛋白反应的样品经HA单抗沉淀后,经Western blot可检测到相应蛋白带。该研究证明了LGALS1与PRRSV 2b蛋白间发生相互作用,为研究PRRSV感染途径和增殖过程提供了理论基础。     

Soil Testing Bray-1 Phosphorus with Non-Reagent Grade Hydrochloric and Sulfuric Acids

Portable instruments have reduced on-site testing costs for soil pH and potassium in isolated locations. Acids could not be shipped to a remote Africa field laboratory. Bray-1 phosphorus (P) testing is difficult because hydrochloric (HCl) is required in the extraction solution and sulfuric acid (H₂SO₄) is used in color development stock solutions. Muriatic HCl acid and battery H₂SO₄ were available in a local stores. The objectives of this study were to evaluate the accuracy of P soil testing using diluted, non-reagent HCl for making modified Bray1-P extractant and non-reagent H₂SO₄ for color development. Experiment 1 was conducted with soil samples from fields using Sunnyside? and Klean Strip? HCl muriatic acids. Soil samples extracted with diluted reagent HCl were closely related to P results using diluted Klean Strip (R² = 0.96) and Sunnyside (R² = 0.84) HCl. In Experiment 2, two commercial brands of H₂SO₄ acid used to refill car battery cells were evaluated. Test results suggested that the battery H₂SO₄ acids are not suitable for making color development solution. In Experiment 3, a small battery powered spectrometer for P testing aquarium water was calibrated for soils. Samples from the Soil Science Society of America –North American Proficiency Testing program (NAPT) were tested with the meter and non-reagent HCl. Fiske-Subbarrow reducing agent and HCl were used in a variation procedure which does not require H₂SO₄ for color development. Eight of 10 samples tested proficient based on NAPT limits (2.5 x Median Absolute Deviation).