Transport functions and expression analysis of vacuolar membrane aquaporins in response to various stresses in rice

The vacuole, a multifunctional organelle of most plant cells, has very important roles in space filling, osmotic adjustment, storage and digestion. Previous researches suggested that aquaporins in the tonoplast were involved in vacuolar functions. The rice genome contains 33 aquaporin genes, 10 of which encode tonoplast intrinsic proteins (TIPs). However, the function of each individual TIP isoform and the integrated function of TIPs under various physiological conditions remain elusive. Here, five rice TIP members were characterized with water and/or glycerol transport activities using the Xenopus oocyte expression system. OsTIP1;2, OsTIP2;2, OsTIP4;1 and OsTIP5;1 possessed water transport activity. OsTIP1;2, OsTIP3;2 and OsTIP4;1 were demonstrated with glycerol transport activity. Rice TIP expression patterns under various abiotic stress conditions including dehydration, high salinity, abscisic acid (ABA) and during seed germination were investigated by real-time PCR. OsTIP1s (OsTIP1;1 and OsTIP1;2) were highly expressed during seed germination, whereas OsTIP3s (OsTIP3;1 and OsTIP3;2) were specifically expressed in mature seeds with a decrease in expression levels upon germination. The results of this research provided a functional and expression profiles of rice TIPs.     

马铃薯StPSKRs基因的克隆及其亚细胞定位分析

该研究采用同源克隆与PCR扩增方法,从马铃薯品种‘Desiree’中克隆植物磺肽素受体基因StPSKR1和StPSKR2的全长cDNA,并对其进行生物信息学分析及亚细胞定位分析,为深入研究StPSKR1和StPSKR2基因在马铃薯生长发育和生物胁迫中的作用提供理论依据。结果发现:(1)通过同源克隆与PCR扩增获得StPSKR1和StPSKR2的全长cDNA片段,并将其克隆到pGWB5-GFP载体;测序结果显示这2个基因编码的蛋白质与数据库给定的蛋白质序列保持一致,表明成功克隆到StPSKR1和StPSKR2基因。(2)StPSKR1位于马铃薯1号染色体上,cDNA全长3 042 bp,编码1 013个氨基酸,预测蛋白相对分子质量为112.16 kD,理论等电点6.27;StPSKR2位于7号染色体,cDNA全长3 135 bp,编码1 044个氨基酸,相对分子量为114.99 kD,理论等电点6.19。(3)生物信息学分析显示,StPSKR1和StPSKR2都属于跨膜蛋白。(4)亚细胞定位结果显示,StPSKR1和StPSKR2均定位于细胞膜上。     

Synthesis and Activity Evaluation of Nasopharyngeal Carcinoma Inhibitors Based on 6‐(Pyrimidin‐4‐yl)‐1H‐indazole

Human nasopharyngeal carcinoma is a common head and neck malignancy with high incidence in Southeast Asia and Southern China. It is necessary to develop safe, effective and inexpensive anticancer agents to improve the therapeutics of patients with nasopharyngeal carcinoma. A series of small molecular compounds based on 6‐(pyrimidin‐4‐yl)‐1H‐indazole were synthesized and evaluated for antiproliferative activities against human nasopharyngeal carcinoma cell lines SUNE1. Compounds 6b , 6c , 6e and 6l showed potent antiproliferative activities similar to positive control drug cisplatin in vitro with lower nephrotoxicity than it. N‐[4‐(1H‐Indazol‐6‐yl)pyrimidin‐2‐yl]benzene‐1,3‐diamine ( 6l ) was selected for further study. It was found that 6l induced mitochondria‐mediated apoptosis and G₂/M phase arrest in SUNE1 cells. Furthermore, compound 6l at 10 mg/kg can suppress the growth of an implanted SUNE1 xenograft with a TGI% (tumor growth inhibition) value of 50 % and did not cause serious side effects in BALB/c nude mice. This study suggests that 6‐(pyrimidin‐4‐yl)‐1H‐indazole derivatives are a series of small molecule compounds with anti‐nasopharyngeal carcinoma activities.     

高山被孢霉淬灭方法及胞内代谢产物的气相色谱-质谱分析比较

在花生四烯酸生产菌高山被孢霉代谢组学研究中,需利用胞内代谢物的提取手段并基于气相色谱-质谱（GC-MS）分析方法对其进行检测。比较了3种胞内代谢物提取方法及不同色谱柱条件下GC-MS分析结果。研究表明：采用冷甲醇淬灭分别较液氮直接淬灭及真空过滤后,减少了胞内代谢物的泄露并更好地实现了胞外及胞内代谢物的分离。在对代谢物分析的比较中,极性色谱柱（DB-FFAP）检出的代谢物仅为11种,主要为有机酸、醛类;而代谢物经衍生化后采用非极性色谱柱（DB-5）共检出32种化合物,主要为糖、糖苷及醇类。     

Enhancing xanthine oxidase fermentation with pH-shift strategy based on kinetic analysis by Arthrobacter M3

The effect of initial culture pH and inducer concentration on xanthine oxidase (XOD) fermentation in shake flasks was first carried out. The results showed that the optimum initial culture pH and inducer concentration were 8.6 and 3.6 g/l, respectively. Batch fermentation of XOD by Arthrobacter M3 in a 7.5-l fermentor was then tested under various pH conditions ranging from 7.6 to 8.6. Based on the analysis of the obtained kinetic parameters, a pH-shift strategy in batch fermentation was implemented to enhance the XOD fermentation. In this strategy, the initial culture pH was set at 8.6 without control and was maintained at 7.6 after the biomass reached 2.0 g/l DCW. XOD production (P) and final average yield coefficient for production on biomass (FAY_p/x) in this strategy reached 7,415.3 U/l and 1,229.7 U/g, respectively, which were significantly higher than the results from the other four protocols. In pH-shift batch fermentation, the Luedeking–Piret equation for product accumulation and the Luedeking–Piret-like equation for substrate consumption fit well with the experimental values. The correlation coefficients (R ²) of these two fitting curves were 0.977 and 0.992, respectively.     

烟粉虱传毒相关蛋白基因的克隆及其表达量分析

【目的】烟粉虱Bemisia tabaci(Gennadius)是一种世界性的入侵性害虫,其传播的番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)给我国番茄生产造成巨大经济损失。为了阐明烟粉虱传播双生病毒的机理,本文拟研究明确B和Q烟粉虱体内传毒相关蛋白Gro EL基因及其表达量。【方法】采用特异性引物克隆了B和Q烟粉虱内共生菌编码的传毒相关蛋白Gro EL基因,并进行序列分析;并利用荧光定量PCR技术检测两种生物型及其获取双生病毒前后该基因的相对表达量。【结果】烟粉虱内共生菌Hamiltonella编码的Gro EL基因全长为1 668 bp,编码555个氨基酸;B、Q烟粉虱该基因的核苷酸序列相似性为99.94%,氨基酸同源性为99.82%;带毒烟粉虱中Gro EL基因的表达量显著高于未带毒的对应生物型烟粉虱;无论带毒与否,Q烟粉虱该基因的表达量均显著高于B(P<0.05)。【结论】烟粉虱携带TYLCV后可诱导Gro EL的表达量升高,B和Q烟粉虱中Gro EL基因及其表达量均存在差异,这可能是B和Q烟粉虱传毒效率存在显著差异的原因之一。     

太子河水体中多环芳烃分布与污染源解析

利用振荡提取-硅胶柱净化-HPLC荧光(FLD)/二级阵列检测器(DAD)检测法测定了太子河水中USEPA16种优控多环芳烃(PAHs)的含量。结果表明,枯水期(4月)、丰水期(7月)和平水期(10月)太子河水中PAHs总浓度分别为454.5～1379.7、1801.6～5868.9和367.0～5794.5ng.L-1,同国内外河流相比,太子河水中PAHs污染较严重,且具有明显的季节分布特征,丰水期PAHs浓度远高于枯水期。丰水期、平水期和枯水期太子河水中均以2～3环PAHs为主,但不同季节代表性PAHs的种类不同。污染来源分析表明,枯水期太子河水中PAHs主要来源于石油污染,丰水期和平水期主要来源于石油源和燃烧源的混合源。     

共转染CDK1、CDK2siRNA对肿瘤细胞周期和细胞凋亡的影响

目的研究共转染CDK1、CDK2siRNA同时抑制CDKI、CDK2蛋白表达对肿瘤细胞周期和细胞凋亡的影响,探讨细胞周期主要调控分子在肿瘤细胞凋亡中的作用。方法以人宫颈癌细胞株HeLa细胞为研究对象,用脂质体lipofectamine2000同时转染CDKl和CDK2siRNA。在转染后48、60h收集细胞,用Western印迹检测CDKl、CDK2蛋白的表达,AnnexinV／PI检测转染细胞的凋亡,流式细胞术DNA含量检测分析细胞周期。转染细胞进行瑞氏一姬姆萨染色（Wright—Giemsa）后在显微镜下观察其形态变化i结果共转染CDKl、CDK2siRNA后48和60h,Western印迹结果显示CDKl和CDK2蛋白的表达都同时降低。共转染CDKl、CDK2siRNA后,细胞周期S期和G1／M期比例与对照相比有明显增加;共转染细胞经瑞氏一姬姆萨染色后在显微镜下可见双核或多核细胞增多;AnnexinV／PI检测结果显示共转染CDK1、CDK2siRNA的细胞在48和60h细胞凋亡率与对照相比有显著的升高。结论siRNA干扰导致的CDKI、CDK2表达同时降低不仅导致细胞周期s期和G1／M期的阻滞,也诱导了肿瘤细胞的凋亡。