马铃薯AP2/ERF转录因子的克隆及植物表达载体构建

AP2/ERF基因家族转录因子普遍存在于植物中,参与植物体内的各种生物学过程,包括植物的生长发育、生物和非生物胁迫响应等。前期转录组测序的结果发现马铃薯‘加湘1号’一个AP2/ERF家族基因(PGSC0003DMG400012154)在接种晚疫病菌(Phytophthora infestans)24 h后被显著激活。从接种P.infestans 24 h的‘加湘1号’的总RNA中通过RT-PCR获得了该基因CDS序列为885 bp,BLAST分析显示该基因编码一个295个氨基酸残基的蛋白,并且含有1个AP2/ERF结构域,是AP2/ERF转录因子家族中的ERF亚家族的一员。本研究将该基因与XcmⅠ酶切的表达载体pCXSN连接,转化大肠杆菌,通过测序挑选插入正确克隆酶切验证,并成功转化农杆菌。本研究结果为进一步研究该基因的功能提供了帮助。     

不同生育期成龄灰枣树茎流速率与气象因子的关系

【目的】在微喷灌的灌溉方式下,研究不同天气和不同气象因子条件下成龄灰枣树的茎流速率,精准测定出成龄枣树的耗水量,为优化枣树的土壤水分管理提供生产指导。【方法】采用针式茎流计,连续监测灰枣树茎流速率,分析枣树的茎流速率受不同天气和气象因子的影响程度。【结果】晴天和阴天茎流速率的波动变化过程呈倒“U”型的单峰曲线,而多云天气下呈“M”型的双峰曲线。1 d中枣树的茎流累积趋势呈“S”变化,各生育期耗水量大小排序为果实膨大期>成熟期>花期>萌芽展叶期。只有成熟期枣树的茎流速率与空气温度相关程度最高,其余都是太阳辐射。各生育期太阳辐射与枣树茎流速率的相关系数R_D1=0.964^**、R_D2=0.969^**、R_D3=0.957^**和R_D4=0.886^**。利用多元线性回归在枣树的各生育期内对茎流速率和气象因子进行输入回归,建立不同天气条件下,各生育期枣树茎流速率和气象因子的多元线性回归模型,经过回归系数和相关系数检验,各生育期的多元线性回归方程均达到了显著水平。【结论】各生育期内,典型天气的茎流速率排序为晴天>多云>阴天,整个生育期中的需水关键期是果实膨大期,对枣树的茎流速率产生最大影响的气象因子是太阳辐射和空气温度。     

Regulatory effect of astragaloside IV on SDF-1α/CXCR4 in EPCs

AIM: To investigate the effects of astragaloside IV (AS-IV) on chemokine receptor 4 (CXCR4) and stromal cell-derived factor 1α (SDF-1α) in endothelial progenitor cells (EPCs) and its mechanism. METHODS: Rat bone marrow-derived EPCs were cultured in vitro. The proliferation, adhesion, migration, apoptosis and tube formation capacity of EPCs treated with AS-IV and AMD3100, a specific blocker of CXCR4, were observed. The effects of AS-IV on the expression of SDF-1α/CXCR4 at mRNA and protein levels and the protein level of p-CXCR4 in the EPCs were determined. RESULTS: AS-IV significantly enhanced the proliferation, adhesion, migration and tube formation abilities of EPCs, reduced the apoptosis of EPCs, and up-regulated the mRNA and protein expression of SDF-1α and CXCR4 and the p-CXCR4 protein level in the EPCs. On the other hand, AMD3100 blocked the up-regulating effect of AS-IV on the mRNA and protein expression of CXCR4 and the p-CXCR4 protein level in the EPCs, but did not affect the effect of AS-IV on the expression of SDF-1α. CONCLUSION: AS-IV might enhance the biological function of EPCs by regulating the expression of SDF-1α/CXCR in EPCs.     

山羊FGF1O基因的克隆及其表达分析

成纤维细胞生长因子10(fibroblast growth factor 10,FGF10)是一种极其重要的生长因子,能促进小鼠(Mus musculus)和人(Homo sapiens)前体脂肪细胞的分化.为了获得山羊(Capra hircus)FGF10基因序列,研究其组织表达特性,确定其在肌内前体脂肪细胞分化过程中的表达模式,并分析FGF10mRNA表达水平与肌内脂肪沉积的关系,本研究以简州大耳羊(C.hircus)为实验动物,采用反转录聚合酶链式反应(RT-PCR)技术克隆FGF10基因,采用Ⅱ型胶原酶消化获得山羊原代肌内前体脂肪细胞,利用实时荧光定量PCR(qPCR)技术检测FGF10在成年山羊不同组织中的表达差异及其在肌内前体脂肪细胞分化过程中的表达水平,并将基因表达水平与肌内脂肪(intramuscular fat,IMF)含量进行关联分析.结果显示,克隆得到山羊FGF10基因(GenBank登录号:KT899958)序列1 252 bp,其中开放阅读框642 bp,编码213个氨基酸,与绵羊(Ovis aries)和牛(Bos taurus)的该基因同源性达100％,具有跨膜结构域和信号肽结构.FGF10 在山羊肺脏中表达水平最高,显著高于其他组织(P＜0.05),在脾脏和脂肪中也存在较高水平的表达.FGF10 mRNA在成年羊背最长肌中表达量显著高于羔羊与育成羊(P＜0.05),相关性分析结果显示,FGF10 mRNA表达量与山羊背最长肌IMF含量呈显著正相关(P＜0.05).FGF10 mRNA在肌内前体脂肪细胞中表达水平最低,在诱导分化后2d达到最高.本研究结果为进一步阐明FGF10基因在山羊肌内脂肪沉积中的分子机制提供了理论依据.     

扎鲁特旗山地草地土壤含水量动态与气候因子的关系

通过对内蒙古通辽市扎鲁特旗巴雅尔吐胡硕牧业气象站1983-2012年气象数据和山地草地土壤含水量数据分析,揭示土壤含水量的动态变化及其与气候因子之间的相关关系。结果表明,1983-2012年山地草地土壤各层(0-10、10-20、20-30、30-40、40-50和0-50cm)含水量均呈现逐渐降低的变化趋势,草地逐渐旱化;土壤含水量下降过程可以分为3个阶段,第1阶段为1983-1987年,该阶段土壤含水量在15%~30%之间波动,属于土壤含水量的较高阶段;第2阶段为1988-1999年,该阶段土壤含水量在5%~15%范围内波动,属于土壤含水量高(15%~30%)向低(2%~10%)过渡阶段;第3阶段是2000-2012年,该阶段土壤含水量在2%~10%范围内波动,属于土壤含水量较低阶段;土壤含水量与温度和日照时数呈负相关,与降水量、相对湿度、蒸散量呈正相关。     

雌雄异株植物生长释放/抑制判定及其气候解释

利用树轮学方法,以长白山雌雄异株植物红豆杉、山杨和水曲柳为研究对象,通过判定雌雄异株树种生长释放/抑制事件及其与气候因子的关系,以期揭示物种属性和性别因素对树木生长释放/抑制事件的影响及其对气候响应机制的差异性。结果表明:3种雌雄植株的生长释放事件大多发生在幼龄阶段,生理特征差异也可能导致不同雌雄植株发生生长释放事件的时间和频率存在一定差异性。性别因素对山杨和红豆杉雌雄植株的生长释放存在显著影响,山杨雌株和红豆杉雄株对外界环境干扰更为敏感,水曲柳雌雄株对环境干扰的响应差异不明显;山杨更易受到外界环境干扰发生生长释放事件;水曲柳植株发生生长释放/抑制事件可能与林窗干扰有关。红豆杉和山杨的生长释放事件与气候因子具有显著相关关系,红豆杉雄株与温度、降水则存在显著正相关关系(P0.05),山杨雄株与温度、降水则存在显著负相关关系(P0.05),山杨雌株与降水呈显著负相关关系(P0.05),植株的生长释放/抑制事件对气候的响应与树木径向生长--逐月气候因子的关系有所不同。因此应当考虑生长--气候因子之间的非线性关系以及物种和性别因素对生长--气候关系的影响,以期完善植株生长对气候的响应机制。      

苦瓜转录因子McEIL2的克隆及其表达分析

本研究从已构建的苦瓜果实均一化文库筛选得到一个EIN3-like同源EST序列,结合RACE技术,克隆得到EIN3基因c DNA全长序列,命名为Mc EIL2(Gen Bank登录号:KF595122)。该基因全长2 122 bp,开放阅读框1 890 bp,编码629个氨基酸,预测该蛋白的分子量为71.58 k D,与黄瓜Cus EIN3、甜瓜Cm EIL1、苜蓿Mt EIL1、绿豆Vr EIL1和番茄Le EIL1的蛋白同源性分别为89.61%、78.37%、71.23%、69.09%和65.66%。Mc EIL2基因的编码蛋白亚细胞定位于细胞核,荧光定量分析表明Mc EIL2基因表达量在幼果期先强后弱,绿熟期最低,破色期表达量大幅增加至最高随后下降,推测该基因参与苦瓜果实成熟软化调控过程。本研究初步明确了Mc EIL2基因在苦瓜成熟过程表达模式,可为今后进一步揭示苦瓜果实成熟软化的分子机制奠定基础。     

Effect of eleutheroside B/E on proliferation and expression of PPARγ and TGF-β1 in HBZY-1 cells cultured with high glucose

AIM: To explore the effects and mechanism of eleutheroside (ETS) B or E on the proliferation of HBZY-1 cells treated with high glucose. METHODS: The HBZY-1 cells were cultured under high glucose condition. The 4th generation of HBZY-1 cells was used for determining the optimal cell density, which was consistent with the growth regulation curve of the cells. The cells were divided into 6 groups: low glucose (LG) group, high glucose (HG) group, high glucose plus ETS-B/E (low dose, medium dose and high dose) groups, and high glucose plus losartan (LTG) group. After all cells were treated with the corresponding drugs at 24 h, 48 h and 72 h, the inhibitory rate of the proliferation was measured, and the expression of TGF-β1 and PPARγ was detected by immunocytochemistry and Western blotting. RESULTS: The best cell density was 2 000 cells/well, which was complied with the basic rules of the cell growth, and high glucose significantly promoted the HBZY-1 cell proliferation. At each time point, the inhibitory effects of ETS-B/E were significantly different between HG group and LTG group on the proliferation of the HBZY-1 cells (P<0.05). The expression of TGF-β1 was significantly inhibited, and the expression of PPARγ was significantly promoted by ETS-B/E (P<0.05). ETS-E showed stronger effect than ETS-B (P<0.05) in a concentration- and time-dependent manner. CONCLUSION: ETS-B/E significantly inhibits the proliferation of HBZY-1 cells under high glucose condition by decreasing TGF-β1 expression and promoting PPARγ expression.     

Detection of Serotypes and Virulence Factors of Escherichia coli from Piglets in Beijing

To investigate the cause of piglets diarrhea, and the distribution of the serotype and virulence factors of swine Escherichia coli in Beijing, 400 diarrhea samples were collected. TSA serum agar culture method was used to isolate Escherichia coli, serotype identification test and virulence factor genes test were used to verify the presence of O-antigen. 64 strains of E.coli were isolated from 400 diarrhea samples, among which 42 strains of E.coli were classified as 8 serotypes:O101(18.7%),O64(12.5%),O8(10.9%),O20(10.9%),O45(4.7%),O149(4.7%),O2(1.6%) and O89(1.6%), and the major virulence factors were STa, Stx2e, astA and eaeA. There were 8 mainly serotypes that caused piglets diarrhea in Beijing area, among which O101 serotype accounted for the highest proportion. The major virulence factors were astA and eaeA, accounted for more than 50% of all strains, STa and Stx2e accounted for more than 30% of all strains. These data would provide effective data to support the prevention and control of piglet diarrhea in Beijing.     

绿豆种质资源农艺性状的因子分析及综合评价

为合理利用绿豆种质资源,本研究以55份绿豆种质资源为材料,通过对其10个农艺性状进行因子分析及聚类分析。结果表明:10个农艺性状的变异系数为8.57%~47.19%,其中单株粒重变异系数最大,生育期变异系数最小。将10个农艺性状简化为结荚数因子、高度因子、粒数因子、粒重因子和生育期因子5个独立的主因子,该5个主因子可以反映原始信息的86.340%。经综合得分排序,评选出10份性状优良的绿豆种质资源。聚类分析将55份种质资源的10个数量性状分为3大类,类群I、类群II、类群III分别包括22份、7份、26份材料。结合综合评价和聚类分析认为,在黑龙江绿豆生产中,‘安绿8号’、‘佳木斯黄绿豆’、‘冀绿1号’可作为品种选育的改良亲本,也可大面积种植利用。