The Effects of TFIBA and IBA on Seminal Root Length and Initiation of Lateral Root Primordial of Rice and Lettuce Seedlings

适宜浓度的TFIBA显著促进了莴苣和水稻初生种子根的伸长生长 ,抑制了水稻种子根侧根原基的发生 ,但对莴苣侧根原基的发生无任何作用 ,对根顶端优势的调控与IBA相反The effects of 4,4,4 trifluoro 3 (indole 3 )butyric acid (TFIBA) and indole 3 butyric acid (IBA) on the growth of primary roots and the genesis of lateral root primordial (LRP) were investigated. It was found that TFIBA at range of 0.001～10 μmol·L -1 inhibited the initiation of LRP while promoted the growth of primary roots of rice seedlings, whose effect was contrary to that of IBA. TFIBA at range of 0.1～100 μmol·L -1 also promoted the growth of lettuce seedlings while did not exhibit any promotive effect on the initiation of LRP

Transformed Rice with Salt Tolerance-related Genes of Bruguiera sexangula by Agrobacterium Meditation

运用农杆菌介导法将红树林耐盐相关基因mangrin转入粳稻品种‘日本晴’中,通过GUS基因检测愈伤组织转化率,确定农杆菌菌液浓度OD600为0.5,浸染时间30min,共培养时间3d为最佳转化体系;经潮霉菌筛选,获得抗性再生植株。通过PCR扩增检测、Southern blot分析和GUS基因活性检测,结果表明,mangrin基因整合到再生水稻的染色体DNA上。耐盐性测定结果表明,转基因植株在200mmol/L NaCl胁迫下,成活率保持在83.3%,株高增长20%~40%,mangrin基因能提高转基因水稻对盐胁迫的抗性。Salt tolerance-related genes in Bruguiera sexangula were transformed into Oryza sativa subsp.japonica cv.Nipponbare,a keng-rice variety.The optimal system of the transformation was found by detecting the callus-transformation rate with GUS gene to be the one with the agro-bacterium concentration set at OD_ 600 0.5,the infection time set at 30 minutes and the culture time set at 3 days and then the regeneration plantlets were obtained by Hyg screening.mangrin gene was integrated in the DNA of the regeneration plantlets by PCR amplification,Southern blotting and GUS gene activity detection.The testing of the salt tolerance indicated that under the stress resulting from 200 mmol/L NaCl the transgenic plants maintained a survival rate of 83.3% and increased their heights by 20%~40% and thus mangrin gene could raise the resistance of the transformed rice to salt stress.中国科学院知识创新工程重要方向项目(KZCX3-SW-444);; 教育部重点项目(01102

The Construction of a Vector for Endosperm-specific Expression of IPT Gene and It's Transformation in Rice

为研究细胞分裂素在水稻籽粒发育中的作用,构建了P1300-P Pg-5α-IPT-nOS植物表达载体,并对水稻进行遗传转化。以水稻品种9311为材料,采用PCr法获得种子中特异表达的Pg-5α基因启动子,并将此启动子与P1300相连接,构建P1300-P Pg-5α载体,用nCOⅠ和SPEⅠ双酶切P Sg516,获得IPT-nOS核酸片段,然后将此核酸片段插入到P1300-P Pg-5α中,构建P1300-P Pg-5α-IPT-nOS植物表达载体。以水稻品种日本晴愈伤组织为材料,采用农杆菌介导的方法进行遗传转化。成功构建了植物表达载体P1300-P Pg-5α-IPT-nOS,获得了阳性率为81.3%转IPT基因群体。获得转基因株系后,将为进一步筛选高效表达株系及观察其籽粒生长发育过程提供基础。In order to specifically express IPT gene in the endosperm of developing rice seed,the binary vector p1300-p PG-5α-IPT-Nos was constructed,firstly,the promoter of PG-5α was obtained by PCR amplification from rice genomic DNA( variety: 9311) and p1300-p PG-5α was constructed by inserting p PG-5α into p1300,then the IPT-Nos fragment was obtained by cutting p SG516 with Nco Ⅰ and Spe Ⅰ.After IPT-Nos inserted into p1300-p PG-5α,p1300-p PG-5α-IPT-Nos was constructed.p PG-5α-IPT-Nos was transformated into Oryza sativa( subsp.japonicacv.Nipponbare) cells by agrobacterium mediated method and regenerative seedlings were obtained.The results of PCR showed that about 81.3% seedlings were positive.唐山师范学院博士基金项目(09A01); 农业部“948”项目(2011-G1(3)-07); 河北省科技计划项目(13396401D

In vitro study of cholesterol succinyl chitosan anchored liposomes as a carrier for epirubicin

背景:多糖"锚定"脂质体在抗肿瘤药物、蛋白以及基因的传输领域有着极其重要的理论及应用价值,国外已有较多机构在进行相关的研究。目的:通过"锚定"的方式制备胆甾醇琥珀酰基壳聚糖锚定脂质体,并以表阿霉素作为模型药物,考察其对包载药物体外释放性质的影响。设计、时间及地点:体外实验,于2006-09/2008-05在天津市生物医学材料重点实验室完成。材料:以壳聚糖为原料,合成胆甾醇琥珀酰基壳聚糖,并采用胶体滴定法测定其胆甾醇基取代度。方法:采用pH梯度法制备表阿霉素脂质体,然后通过共孵育的方法合成了取代度为2.80%,5.58%和8.00%的载药胆甾醇琥珀酰基壳聚糖锚定脂质体。主要观察指标:荧光分光光度计检测药物浓度;透射电镜观察脂质体形态;亚微米粒度及电位分析仪检测脂质体的粒径大小、分布和电位;动态透析法考察包载药物表阿霉素在胆甾醇琥珀酰基壳聚糖锚定脂质体中的体外释放特征。结果:胆甾醇琥珀酰基壳聚糖锚定脂质体为规则球状形态,呈现典型的核壳结构,粒径为245.4~279.7nm,zeta电位为+21.09~+25.48mV;和载药脂质体及壳聚糖包衣脂质体相比,CHCS锚定脂质体能明显延缓表阿霉素的体外释放,在胆甾醇基取... 【英文摘要】 BACKGROUND: Polysaccharides anchored liposomes play an extremely important role in the fields of antitumor drug, protein and gene transmission. Related research is also present abroad. OBJECTIVE: To prepare cholesterol succinyl chitosan (CHCS) anchored liposomes, and to investigate the effect of CHCS anchored liposomes on the release property in vitro of loading drugs taking epirubicin as a model drug. DESIGN, TIME AND SETTING: A study in vitro was performed in the Key Laboratory of Biomedical Materials o

Proteomic Analysis of Rice Cultivar Jiafuzhan in the Responses to Xanthomonas campestris pv.oryzicola Infection

作者简介: 陈芳育(1978-) , 男, 讲师。E-mail : cfy307@ sohu. com * 通讯作者(Corresponding author) : 陈亮( 1963-) , 男, 教授, 博士生导师, 研究方向: 细胞与分子生物学。E-mail: chenlg@ xmu. edu. cn[中文文摘]运用双向电泳分析高抗水稻品种“佳辐占”受强毒力细菌性条斑病病原菌侵染2d后的叶片蛋白质组变化,共发现38个蛋白质发生差异表达,其中32个上调,5个下调,1个新增。用MALDI-TOF-MS分析和数据库检索鉴定出其中的33个差异表达蛋白质,并将它们分为4个功能类群,即信号转导相关蛋白、防卫相关蛋白、代谢相关蛋白和蛋白质稳定相关蛋白。这些蛋白分别参与了信号识别、信号传递、抗氧化、糖代谢、细胞壁加固、植保素合成等抗病生理反应。研究表明,水稻对细菌性条斑病病原菌的侵染存在着一个复杂的抗病信号应答和代谢调控网络,其作用机理可以通过差异表达的蛋白质(酶)反映出来,其中差异表达的8个R蛋白和3个PR蛋白可能与水稻对细菌性条斑病的抗病性密切相关。本研究为进一步揭示水稻对细菌性条斑病的抗性机理及相关抗病基因的功能克隆提供了依据。[英文文摘]Rice bacterial leaf streak( BLS) caused by the pathogen Xanthomonas oryzae pv. oryzicola ( Xooc) is one of the major rice diseases in South China. Here we focus on proteomics as a tool for the discovery of differentially expressed proteins closely related to the disease resistance. The leaves of rice cultivar Jiafuzhan (Oryzae sativa L. ) highly resistant to the disease, were infected by"89773-1- 1" strain of the Xooc with strong pathogenicity. Total proteins were extracted from the leaves sampled at two days after inoculation, and separated by two- dimensional electrophoresis. It was found that there were thirty- eight proteins expressed differentially, of which thirty-two were up-regulated, five down-regulated and one was "new". Of the thirty- eight responsive proteins, thirty-three were identified by MALD-I TOF-MS and database searching.Based on the predicted function, we grouped them into four clusters: signal transduction, defensive responses, substance metabolism and protein stabilization, which were involved in many resistant physiological react ions, including signal recognition and transduction, antioxidant react ion, carbonhydrate metabolism, cel-l wall reinforcement and phytoalexin biosythesis. In turn a complex signal transduct ion and metabolic regulative network in the resistant responses to the infection of Xooc was outlined in this work, and the molecular mechanism was revealed by differentially expressed protein/enzyme patterns during Xooc infection. In this study, eight R proteins and three pathogenesis- related(PR) proteins which might relate closely to the disease-resistance were found. This result provides us the basic information to further reveal the resistant mechanism and conduct functional cloning of the resistan-t related genes in rice to BLS.生物农药与化学生物学教育部重点实验室( 福建农林大学) 开放课题基金项目( KF0411

黄河游荡河段河床形态调整对洪水过程的响应

以黄河流域1950~1985年200余场洪水资料为基础,并增加了最近的实验资料,分析了黄河下游游荡河段不同含沙量洪水过程中河床形态的调整过程。结果表明,由洪水过程所导致的河床形态变化是相当剧烈的,且与含沙量密切相关,表现出非线性的变化规律。当含沙量较小时,随含沙量的增大,洪水后河床宽深比增大,当含沙量增大到一定程度后再增大时,宽深比随含沙量的增大而减小。这一结果为修正Schumm关于河床形态变化的定性预测关系提供了新的依据

Cloning salt-tolerance related gene ZRP4 of rice and construction of its plant expression vector

采用RT-PCR技术克隆了水稻O-甲基转移酶ZRP 4基因的cDNA片段,并进行了序列测定与分析。结果表明,该序列全长1 101 bp,编码366个氨基酸,与G enB ank中水稻ZRP 4基因序列比对,核苷酸同源性为99.8%,氨基酸同源性为99.7%。将克隆片段插入中间载体pBPFΩ7,经H indⅢ酶切回收带有P 35s启动子和nos终止子片段,连接pCAM B IA 1301载体,成功构建ZRP 4基因的植物表达载体。Utilizing RT-PCR,the cDNA of fragment of O-methlytransferanse ZRP4 gene in rice was successfully amplified,which was about 1 101 bp in length and encoded 366 amino acids,and was cloned and sequenced.DNA sequence analysis indicated that the cloned fragment shared high homogeneity to the cDNA of ZRP4 gene in rice.The cloned cDNA of ZRP4 gene was introduced into a middle vector pBPFΩ7.The fragment with P35s promoter and nos were digested by Hind Ⅲ,which were ligated into pCAMBIA1301 vector,giving an plant expression vector containing ZRP4 gene.中国科学院知识创新工程重要方向项目(KZCX3-SW-444);; 教育部重点项目“01102-培育转耐盐基因水稻的研究

风水两相变化对黄河中游支流粗泥沙的影响

基于黄河中游河口-龙门区间实地考察和对1964—1989年的实测资料进行时空变化分析,结果发现:该区水力、风力变化对河流产出的粗泥沙的量和质的变化具有一定的分异控制作用,水力对河流的输沙量的变化起主导作用,而风力对粗泥沙占全沙的百分比变化起主导作用。随着降雨量的减少和风沙活动的减弱,黄河中游河流输沙量和粗沙比有不同程度的减少,输沙量减少明显,减沙比在50%～70%之间,而粗沙比的变化要小得多,减少量在10%～20%之间

Cloning and Expression of Vibrio cholerae CTB Gene and the Recombinant CTB Protein Activation Assay

霍乱弧菌CTB蛋白具有免疫佐剂活性。根据已发表CTB基因的序列设计一对引物,从一株霍乱弧菌中扩增出CTB基因,测序后发现该基因全长375bp,与国内分离的六株CTB基因的同源性达96.0%~99.2%。将该基因与pTWIN1连接构建了原核表达载体pTWIN1-CTB,重组表达载体转化BL21(DE3)表达菌株,0.8mmol/LIPTG诱导4h后,收获的细菌总蛋白SDS-PAGE电泳显示CTB在原核表达系统中得到表达,融合蛋白大小与理论值符合,蛋白产量占细菌总蛋白的20%左右,主要以包涵体形式存在,Western杂交和GM1-ELISA结果表明重组蛋白具有免疫原性和粘膜佐剂活性。CTB protein possessed mucosal adjuvant immunoactivity. The CTB gene was amplified by PCR method from a strain V. cholerae. The nucleotide sequence of CTB gene was 375 bp and shared 96.0%~99.2% homology with other 6 CTB genes. The recombinant plasmid pTWIN1-CTB transformed E. coli strain BL21(DE3) expressed with 0.8 mmol/L IPTG. The molecular weight of expression products was identical with expectative weight by SDS-PAGE electrophoresis. The CTB fusion proteins mainly assembled inclusion bodies and the outputs of proteins were approximately 20% of the total bacterial proteins. The CTB proteins possessed mucosal immunoactivity by GM1-ELISA assay.福建省科技厅重点项目(2003N051);; 福建省科技厅配套项目(JY0B3020

Construction of OsICK1 Plant Antisense Expression Vector

O sICK1是水稻细胞周期蛋白依赖性激酶抑制剂基因.我们构建了水稻细胞周期蛋白依赖性激酶抑制剂基因(O s-ICK1)的植物反义表达载体,在农杆菌的介导下尝试使antisenseO sICK1整合到水稻基因组中并初步得到验证.为进一步研究O sICK1对水稻细胞周期调控的作用奠定了基础.Cyclin-dependent protein kinases(CDKs) have a central role in cell cycle regulation.It can be inhibited by the binding of small protein CDK inhibitors(ICKs).OsICK1 is a putative cyclin-dependent protein kinase inhibitor gene found in rice.The aim of this research was to study the fuction of OsICK1 by expressing antisence OsICK1 in rice.First,we constructed a plant OsICK1 antisense expression vector under the control of CaMV35S promotor and NOS terminator.Then,the antisense gene was integrated into the genome of rice callus via Agrobacterium tumefaciens EHA105 mediation.It provided a reliable base for the function study on OsICK1