Proteomic approach to the different expressed proteins induced by infection of Xanthomonas oryzae pv. oryzicola in rice cultivar minghui63

水稻细菌性条斑病（简称细条病），广泛分布于华南、中南稻区。该病是细条病菌Xanthomonasoryzaepv.oryzicola(Xooc)引起的一种细菌性病害，于20世纪50年代在我国广东省首先发现，后因感病的杂交水稻大面积推广，致使此病迅速蔓延，病区不断扩大，已成为我国水稻主要病害和重要的检疫对象。目前国内外对于水稻细条病的研究仅仅限于基因组学的研究以及抗性基因的转化，而对该病菌侵染水稻后诱导机体蛋白应答的系统性研究还未见报导。因此，运用差异蛋白质组学手段开展水稻对细条病菌侵染的应答相关蛋白研究，无论是在阐明水稻的抗性分子机理和基因表达调控方面，还是在高产、优质、高抗的水稻品种的分子育种...Rice bacterial leaf streak (BLS) distributes widely in South China. BLS which was caused by Xanthomonas oryzae pv. oryzicola (Xooc) firstly discovered in the 1950’s in Guangdong, and then spread quickly because the infective hybridized rice was popularized. So BLS is one of the major rice diseases in South China and becomes an important subject of quarantine in our country. Presently, the research...学位：理学硕士院系专业：生命科学学院生物化学与生物技术系_细胞生物学学号：20032605

三色(Bougainvillea peruviana ‘Thimma’)在转录水平的甜菜色素和类黄酮积累比较（英文）

Bougainvillea peruviana‘Thimma’属于三角梅属,该属植物积累甜菜色素而不是像绝大多数高等植物一样积累花青素。该材料特征同株出现3种颜色:白色、洋红色和白/洋红相间。本研究首次使用3种花色特征的花序(红色Yp、混合色的Ym、白色Yw)作为研究材料进行高通量测序。并通过real-time PCR方法对探测到的花色代谢基因进行验证。共获得平均长度为616 bp的73 325条基因。3种材料的差异显示基因(DEGs)中有327个被注释到甜菜色素合成基因,308个被注释到类黄酮合成基因,466个被注释到花青素合成基因。我们选出8个基因:4个甜菜色素合成基因(PPO,CYP76AD1,c DOPA-5-GT,DODA)和4个花青素合成基因(FLS,DFR,LDOX,3-GT)进行验证。其中,4个甜菜色素合成基因在3种花色材料中的表达较好的正相关于甜菜色素含量。花青素合成途径末端的3个基因(DFR,LDOX,3-GT)在B.peruviana中首次被验证。real-time PCR的验证结果很好的吻合转录组测序的结果。同时,B.peruviana也提供了一个很好的三角梅属植物的生理、生化和分子生物学研究的工具,有效的摒除其他生物学干扰。The National Natural Science Foundation of China(Grant No.31071814；31372093)；Xiamen Creation Plantform of Science&Technology(3502Z20131004)

Research on Salt Tolerance of Rice Transformed by mangrin Gene

以转MAngrIn基因水稻的阳性植株(11#,59#,86#)和阴性植株为材料,在0,100,150,200 MMOl/l的nACl溶液中经盐胁迫7 d后,测定叶片相对含水量、质膜透性、超氧化物歧化酶(SOd)活性和脯氨酸含量4项耐盐指标。结果表明:盐胁迫下,转基因植株的叶片相对含水量、脯氨酸含量、SOd活性明显高于阴性植株,叶片电导率则低于阴性植株。对转基因植株和阴性植株的耐盐性综合评定结果是11#>59#>86#>阴性植株。Plants transformed by mangrin gene(11#,59#,86#) and negatives were tested using several salt tolerance indices,including related water content,cell membrane permeability,SOD activity and proline content,which were conducted under the NaCl concentration of 0,100,150,200 mmol/L after 7 days.The results indicated that the transgenic rice plants accumulated more RWC,proline content and SOD activity than ck under salt stress conditions,yet cell membrane permeability decreased less than ck.According to physiological target in synthetically evaluation,the result is 11#> 59#> 86#> Negatives.福建省青年人才基金项目(2006F3022);福建省自然科学基金项目(2007J0250

盐生植物盐角草对中肋骨条藻的生长抑制

利用盐生植物抑制藻类生长是防治海洋赤潮发生的一种新兴技术.本文以中国近海典型赤潮原因种中肋骨条藻为对象,研究了滩涂盐生植物盐角草对藻生长的影响及其机制,并利用高效液相色谱技术分析潜在的抑藻物质.共培养实验结果显示,在营养盐充足的条件下,盐角草能够显著抑制中肋骨条藻的生长,降低藻生长水体叶绿素A(CHlA)浓度.除盐角草根系水提物之外,其余3种有机相提取物均能抑制藻的生长,改变藻细胞大小,并且影响单个细胞的CHlA含量.各种有机相提取物的抑制作用具有浓度效应,随浓度升高抑制作用增强;以甲醇相提取物效果最明显,其次是正丁醇和氯仿相提取物.经高效液相色谱分析,在甲醇相提取物中鉴定出2种黄酮化合物——芦丁和槲皮素-3-β-d-葡萄糖甙.其中芦丁对中肋骨条藻的生长具有一定的抑制作用,其抑制效果随芦丁浓度的升高而增强,随藻细胞初始接种密度的升高而减弱.结果表明,盐角草能够抑制中肋骨条藻的生长,芦丁作为盐角草的代谢产物之一有望成为新的抑藻剂.中科院知识创新重要方向项目(批准号:KSCX2-EW-J-1);国家高新技术研究发展计划(批准号:2007AA091704)资

嗜虫耶尔森氏菌HlyA及HasA外分泌表达系统的构建

【目的】构建一株具备外分泌蛋白功能的工程菌,解决杀虫毒素无法由胞内分泌至胞外,无法直接作用于虫体等问题,为松墨天牛防治提供新思路。【方法】本研究先测定从松墨天牛肠道及其生境中分离出的嗜虫耶尔森氏菌(CSLH88)的生长特性及抗性,进而对其进行分子改造。构建HlyA (pGHKW2)以及HasA (pGHKW4)外分泌表达载体,利用电穿孔法将其转入CSLH88菌株,获得能够表达绿色荧光蛋白的工程菌。利用稀释涂板及荧光体式镜检测技术对两个质粒进行遗传稳定性检测,并采用SDS-PAGE及Western blotting技术验证蛋白外分泌功能。【结果】CSLH88菌株培养2–4 h能够进入对数生长期,并对卡那霉素(Kan)敏感。成功构建了含有Kan抗性基因的pGHKW2(GenBank:MK562405)和pGHKW4(GenBank:MK562404)两个外分泌表达载体的CSLH88工程菌株。其中,发现pGHKW4质粒更加适合在嗜虫耶尔森氏菌中稳定遗传。SDS-PAGE及Western blotting检测结果表明HlyA系统无法在CSLH88菌株中将目的蛋白分泌到胞外,而HasA系统则可以有效地发挥外分泌表达功能。【结论】通过对HlyA及HasA两个外分泌表达系统进行研究,从中筛选出HasA型血红素转运系统作为CSLH88菌株的外分泌表达系统,为后续外分泌杀虫毒素蛋白菌株构建以及CSLH88菌株的致病性研究奠定基础。国家重点研发计划课题(2017YFD0600105);;\n国家自然科学基金(31601905);;\n福建省科技厅自然科学基金(2016J01097);;\n福建省科技计划项目(2018N5002);;\n福建农林大学科研基金(xjq201614);福建农林大学林学院林学高峰学科项目(71201800720,71201800753,71201800779);;\n福建省大学生创新创业训练计划项目(201810389102);;\n福建省林业科学研究项目[Minlinke(2017)03]~

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

Analysis of the Defense Responsiveness in Hybrid Rice Minghui63 Against Bacterial Leaf Streak(Xanthomonas oryzae pv.Oryzicola) Using Two-dimensional Electrophoresis and Mass Spectrometry

水稻细菌性条斑病是由细条病菌Xanthom onas oryzaepv.oryzicola(Xooc)引起的一种细菌性病害,是我国南方稻区的主要病害.为了探明水稻对细条病的应答机制,我们应用双向电泳联用质谱技术,研究了水稻品种明恢63在水稻细条病病菌侵染后不同时期的全蛋白变化,并对差异蛋白进行了鉴定以及分析归类.共鉴定出17种表达上调的蛋白和10种表达下调的蛋白,这些蛋白参与了水稻对细条病侵染的信号识别及防卫应答,其中包括信号转导类蛋白、防卫相关蛋白、代谢相关蛋白和参与蛋白质合成的蛋白等.Rice bacterial leaf streak(BLS) caused by pathogen Xanthomonas oryzae pv.oryzicola(Xooc) is one of the major rice diseases in South China.For proving up the defense mechanism of rice against Bacterial Leaf Streak,the differentially express proteins of Minghui 63 leaf after BLS infection in different inoculation time were analysised by using the two-dimensional electrophoresis and mass spectrometry.17 up-regulated proteins and 10 down-regulated proteins were identified.These proteins involvd in signal recognition and defense response,including signal transduce proteins,defense associated proteins,metabolism associated proteins and protein synthesis associated proteins.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(KF0411)资

硅磷酸铝分子筛SAPO-34稳定性的研究

利用差热分析（ＤＴＡ）、原位ＸＲＤ技术研究了ＳＡＰＯ－３４分子筛原粉样品的晶体结构在模板剂烧除过程以及环境气氛中的变化情况。采用１０７３Ｋ下的高温焙烧和水蒸汽处理以及甲醇转化试验跟踪考察了该分子筛在结构和催化性能上的稳定性

阵列CsI(Tl)探测器鉴别轻带电粒子

文中描述了一个5×5阵列CsI(Tl)探测器结构以及对轻带电粒子的鉴别,CSI晶体有快慢两种成分。通过不同延迟和积分门的选择得到这两种信号,来进行轻带电粒子的鉴别。经过理论模拟和实验测试,发现不同的延迟和门的宽度对鉴别能力都有影响