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Seedling Petri-dish inoculation method:A robust,easy-to-use and reliable assay for studying plant-Ralstonia solanacearum interactions 被引量:1
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作者 CAO Peng CHEN Jia-lan +6 位作者 LI Ning-ning ZHANG Shuang-xi WANG Rong-bo LI Ben-jin LIU Pei-qing AN Yu-yan ZHANG Mei-xiang 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2023年第12期3709-3719,共11页
Ralstonia solanacearum causes a lethal bacterial wilt disease in many crops,leading to huge losses in crop production every year.Understanding of plant-R.solanacearum interactions will aid to develop efficient strateg... Ralstonia solanacearum causes a lethal bacterial wilt disease in many crops,leading to huge losses in crop production every year.Understanding of plant-R.solanacearum interactions will aid to develop efficient strategies to control the disease.As a soilborne pathogen,R.solanacearum naturally infects plants via roots.A huge limitation in studying plant-R.solanacearum interactions is the large variation of R.solanacearum infection assay due to the variable soil conditions and uneven inoculum exposure.Here,we developed a robust and reliable Petri-dish inoculation method which allows consistent and stable infection in young plant seedlings.This method is easy to use,takes about only 10 days from seed germination to the completion of inoculation assay,and requires less inoculum of bacteria as well as growth chamber space.We proved the efficacy of the seedling Petri-dish inoculation method by analyzing plant defense primed by molecular patterns,resistance of defense-related plant mutants,and virulence of R.solanacearum mutants.Furthermore,we demonstrated that the seedling Petri-dish inoculation method can be applied to other host plants such as tobacco and has great potential for high-throughput screening of resistant plant germplasms to bacterial wilt in the future. 展开更多
关键词 bacterial wilt ralstonia solanacearum Petri-dish inoculation method VIRULENCE resistance defense priming
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Children with infectious pneumonia caused by Ralstonia insidiosa:A case report 被引量:1
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作者 Shuang-Zhu Lin Mei-Jia Qian +7 位作者 Yan-Wei Wang Qian-Dui Chen Wan-Qi Wang Jia-Yi Li Rui-Tong Yang Xin-Yao Wang Chun-Yu Mu Kai Jiang 《World Journal of Clinical Cases》 SCIE 2023年第9期2002-2008,共7页
BACKGROUND Ralstonia is a Gram-negative non-fermentative bacterium widespread in nature,and includes four species,Ralstonia pickettii,Ralstonia solanacearum,Ralstonia mannitolilytica,and Ralstonia insidiosa,which were... BACKGROUND Ralstonia is a Gram-negative non-fermentative bacterium widespread in nature,and includes four species,Ralstonia pickettii,Ralstonia solanacearum,Ralstonia mannitolilytica,and Ralstonia insidiosa,which were proposed in 2003.Ralstonia is mainly found in the external water environment,including municipal and medical water purification systems.This bacterium has low toxicity and is a conditional pathogen.It has been reported in recent years that infections due to Ralstonia are increasing.Previous studies have shown that most cases of infection are caused by Ralstonia pickettii,a few by Ralstonia mannitolilytica,and infections caused by Ralstonia insidiosa are rare.CASE SUMMARY A 2-year-old Chinese child suffered from intermittent fever and cough for 20 d and was admitted to hospital with bronchial pneumonia.Bronchoscopy and alveolar lavage fluid culture confirmed Ralstonia insidiosa pneumonia.The infection was well controlled after treatment with meropenem and azithromycin.CONCLUSION Ralstonia infections are increasing,and we report a rare case of Ralstonia insidiosa infection in a child.Clinicians should be vigilant about Ralstonia infections. 展开更多
关键词 CHILDREN INFECTIONS ralstonia insidiosa PNEUMONIA TREATMENT Case report
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Ralstonia metallidurans CH34苯酚降解特性的研究 被引量:11
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作者 高振贤 马宏 +2 位作者 贾振华 李承光 宋水山 《微生物学通报》 CAS CSCD 北大核心 2005年第1期44-47,共4页
RalstoniametalliduransCH34是从一家锌工厂的沉积物中分离筛选到的一株细菌。对其降解苯酚的特性进行了研究。结果表明R.metalliduransCH34具有很高的降解苯酚的能力,其降解苯酚的速率常数为0.33,降解苯酚的最适条件为pH7.0,温度30℃,... RalstoniametalliduransCH34是从一家锌工厂的沉积物中分离筛选到的一株细菌。对其降解苯酚的特性进行了研究。结果表明R.metalliduransCH34具有很高的降解苯酚的能力,其降解苯酚的速率常数为0.33,降解苯酚的最适条件为pH7.0,温度30℃,装液量20%(v/v)。在高浓度重金属存在的条件下,R.metalliduransCH34仍保持较高的苯酚降解活力。柠檬酸钠、琥珀酸则能促进其对苯酚的降解。 展开更多
关键词 ralstonia metallidurans CH34 苯酚 生物降解 重金属
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植物病原细菌Ralstonia solanacearum GMI1000中分泌蛋白信号肽分析 被引量:12
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作者 黄俊丽 吴金钟 +2 位作者 肖崇刚 李常军 王贵学 《遗传》 CAS CSCD 北大核心 2007年第11期1409-1416,共8页
利用SignalP3.0、TMHMM2.0、TargetP1.01、LipoP1.0和PSORTb蛋白分析软件并结合L值计算,对植物病原细菌Ralstonia solanacearum GMI1000菌株基因组中的全部3440个ORFs进行了分析预测,确定其中186个ORFs所编码蛋白质的N-端有信号肽序列,... 利用SignalP3.0、TMHMM2.0、TargetP1.01、LipoP1.0和PSORTb蛋白分析软件并结合L值计算,对植物病原细菌Ralstonia solanacearum GMI1000菌株基因组中的全部3440个ORFs进行了分析预测,确定其中186个ORFs所编码蛋白质的N-端有信号肽序列,且它们的氨基酸残基相对保守。其中134条具有分泌型信号肽,22条具有RR-motif型信号肽,30条具有信号肽酶Ⅱ型信号肽。对各类信号肽及其结构域的长度作了系统的分析。未发现Prepilin-like信号肽和细菌素和信息素信号肽。 展开更多
关键词 ralstonia SOLANACEARUM 基因组 信号肽 蛋白分析
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Ralstonia sp. strain U2菌株对萘的趋化作用 被引量:4
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作者 辛玉峰 刘虹 +1 位作者 王淑君 周宁一 《应用与环境生物学报》 CAS CSCD 北大核心 2005年第4期490-492,共3页
一株降解萘的细菌Ralstonia sp. strain U2对萘具有趋化现象,萘、水杨酸都可以作为其对萘产生趋化现象的诱导物.U2菌对水杨酸、龙胆酸同样都具有趋化性,但这种趋化性是组成型的,不需要诱导.
关键词 趋化 ralstonia sp. STRAIN U2
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应用计算机手段分析植物病原细菌Ralstonia solanacearum的蛋白质序列 被引量:4
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作者 刘雅婷 李成云 +1 位作者 李永忠 李正跃 《云南农业大学学报》 CAS CSCD 2007年第4期485-490,共6页
应用SignalP 3.0,LipoP和TargetP对植物病原细菌Ralstonia solanacearum基因组中的3440个ORFs(openreading frames)进行了信号肽分析,同时系统分析了信号肽的类型及结构。结果表明,3440个ORFs中有462个ORFs所编码蛋白质具有N-端有信号... 应用SignalP 3.0,LipoP和TargetP对植物病原细菌Ralstonia solanacearum基因组中的3440个ORFs(openreading frames)进行了信号肽分析,同时系统分析了信号肽的类型及结构。结果表明,3440个ORFs中有462个ORFs所编码蛋白质具有N-端有信号肽序列,其中348个分泌类信号肽、100个信号肽具有RR-motif信号肽,14个脂蛋白类信号肽,未发现Prepilin-like信号肽和Bacteriocin and Pheromone信号肽。在这462个具有可切割信号肽的分泌蛋白中,有84.0%蛋白质为胞外分泌型(S型),13.2%为线粒体分泌型(M型),另外有2.8%为其它类型的分泌蛋白。通过LipoP分析该菌的全基因组预测具有4种类型蛋白质,其中其中SpI有425个,占12.3%;SpII有80个,占2.3%;CYT有2 541个,占73.9%;TMH有414个,占12.0%。比较了Ralstonia solanacearum和Pseudomonas syringaepv.tomato信号肽的长度及氨基酸的组成,发现这两种菌在这些方面存在这很大程度的相似性。本研究通过对Ralstonia solanacearum进行分泌蛋白和信号肽结构的分析,为将来功能基因组和分泌型外源蛋白的利用提供理论基础。 展开更多
关键词 ralstonia SOLANACEARUM ORF 信号肽 RR—motif 脂蛋白 SignalP 3.0 LipoP TargetP
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Ralstonia metallidurans CH34苯酚、甲苯降解基因的生物信息学分析 被引量:3
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作者 宋水山 段普凡 +3 位作者 贾振华 高振贤 马宏 张小兵 《河北省科学院学报》 CAS 2003年第4期245-251,262,共8页
 采用生物信息学方法对RalstoniametalliduransCH34基因组序列进行了深入分析,推测了参与降解苯酚、甲苯/二甲苯降解的有关基因及功能,编码序列推测结果显示,R.metalliduransCH34中可能存在苯酚、甲苯/二甲苯间位降解代谢途径的基因操...  采用生物信息学方法对RalstoniametalliduransCH34基因组序列进行了深入分析,推测了参与降解苯酚、甲苯/二甲苯降解的有关基因及功能,编码序列推测结果显示,R.metalliduransCH34中可能存在苯酚、甲苯/二甲苯间位降解代谢途径的基因操纵子及其调节因子和辅助因子,为后续进一步克隆和研究各基因的准确功能提供了有益的信息,奠定了基础。 展开更多
关键词 ralstonia metallidurans CH34 苯酚 甲苯 降解 基因 生物信息学 革兰氏阴性菌 废水处理
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植物病原细菌(Ralstonia solanacearum)染色体基因组分泌信号肽计算机分析 被引量:1
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作者 周晓罡 丁玉梅 +2 位作者 张绍松 孙茂林 李建平 《西南农业学报》 CSCD 2008年第2期340-345,共6页
应用SignalP 3.0、TMHMM 2.0、THUMBUP、big-PI、TargetP1.01、Lipop 1.0、TatP 1.0等7种软件对植物病原细菌Ralstonia solanacearum GMI1000染色体基因组3448个氨基酸序列进行预测分析。结果表明,该物种染色体基因组分泌信号肽ORFs有17... 应用SignalP 3.0、TMHMM 2.0、THUMBUP、big-PI、TargetP1.01、Lipop 1.0、TatP 1.0等7种软件对植物病原细菌Ralstonia solanacearum GMI1000染色体基因组3448个氨基酸序列进行预测分析。结果表明,该物种染色体基因组分泌信号肽ORFs有178个,占其基因组编码ORFs的5.2%,其中有150个ORFs属于Ⅰ型分泌信号肽,28个ORFs属于Ⅱ型分泌信号肽。在178个ORFs中具RR-motif信号肽结构的有13个,其中11个属于Ⅰ型信号肽,2个属于Ⅱ型信号肽。通过软件预测未发现Comc型信号肽与细菌素-信息素型信号肽结构。在染色体分泌信号肽ORFs中,有116个具可预测功能,其余62个为功能未知的推定蛋白。已知功能主要集中于细胞代谢、细胞调控及转运、细胞膜结构等领域,这些功能是该物种在长期进化过程中与环境中各种因子发生互作,相互适应的结果。 展开更多
关键词 ralstonia SOLANACEARUM GMI1000 染色体基因组 分泌信号肽
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Ralstonia eutropha菌株镉抗性调节基因CzcR的克隆与序列测定 被引量:2
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作者 段学军 黄春晓 《生态环境》 CSCD 北大核心 2007年第6期1665-1668,共4页
G-细菌镉抗性决定子是一个编码4个蛋白的CzcCBAD操纵子,参照GenBank已登录Czc基因序列进行引物设计,利用PCR技术,从可在含350mg/LCdCl2的培养基上生长的抗镉菌株Ralstoniaeutropha质粒中,扩增出长度约为1200bp的革兰氏阴性细菌镉抗性系... G-细菌镉抗性决定子是一个编码4个蛋白的CzcCBAD操纵子,参照GenBank已登录Czc基因序列进行引物设计,利用PCR技术,从可在含350mg/LCdCl2的培养基上生长的抗镉菌株Ralstoniaeutropha质粒中,扩增出长度约为1200bp的革兰氏阴性细菌镉抗性系统中的抗性调节基因CzcR,然后将其亚克隆到pGEM-T-easy载体上,并转化至受体菌中,构建重组质粒,采用碱性裂解法提取质粒DNA后,经EcoRI酶切分析和核苷酸序列分析,其与GenBank中登录的CzcR基因序列相似性高达98%,显示其具有正确的CzcR基因核苷酸序列。镉抗性基因的获得将大大推动对微生物抗重金属机理的研究,并为进一步构建耐镉基因工程菌,高效净化重金属污染环境打下坚实的基础。 展开更多
关键词 ralstonia eutropha CzcR基因 克隆 序列测定
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高密度Ralstonia eutropha细胞培养过程中二氧化碳的生成及其抑制作用 被引量:1
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作者 尚龙安 范代娣 Ho Nam Chang 《高校化学工程学报》 EI CAS CSCD 北大核心 2004年第5期633-637,共5页
测定了高密度R. eutropha细胞培养过程中二氧化碳的生成量。当通气量维持在1.0vvm时,出口气体中二氧化碳的最大浓度高达15%。为了了解高密度细胞培养过程中二氧化碳对细胞生长代谢可能产生的抑制作用,通过施加不同浓度、不同持续时间的... 测定了高密度R. eutropha细胞培养过程中二氧化碳的生成量。当通气量维持在1.0vvm时,出口气体中二氧化碳的最大浓度高达15%。为了了解高密度细胞培养过程中二氧化碳对细胞生长代谢可能产生的抑制作用,通过施加不同浓度、不同持续时间的二氧化碳脉冲,模拟测试了5升发酵罐内二氧化碳对R. eutropha细胞生长代谢的抑制作用。实验发现,二氧化碳对细胞生长的抑制作用随着二氧化碳脉冲时间的增长和浓度的增高而增强。当在细胞指数生长初期施加6小时浓度为22%的二氧化碳脉冲,最终细胞浓度降低了31%。 展开更多
关键词 高密度细胞培养 二氧化碳抑制作用 二氧化碳生成 ralstonia eutropha
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Ralstonia eutropha CH34酯酶基因在大肠杆菌中的表达及酶学特性
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作者 宋水山 贾振华 高振贤 《河北省科学院学报》 CAS 2001年第1期39-42,共4页
从RalstoniaeutrophaCH34基因文库中筛选出编码酯酶EstA的基因 ,将其克隆于大肠杆菌可以得到稳定表达 ,表达产物具有较高的酯酶活性。对重组酯酶的各种酶学性质进行的研究表明 ,酯酶EstA的最适反应 pH为 7.0 ,其最适反应温度为 5 0~ 80... 从RalstoniaeutrophaCH34基因文库中筛选出编码酯酶EstA的基因 ,将其克隆于大肠杆菌可以得到稳定表达 ,表达产物具有较高的酯酶活性。对重组酯酶的各种酶学性质进行的研究表明 ,酯酶EstA的最适反应 pH为 7.0 ,其最适反应温度为 5 0~ 80℃ ,该酶对酸碱环境适应性较强。在 pH4~ 10 .5范围内保持稳定 ,而在 0~ 6 0℃范围内保温 2h ,该酶仍能保持 80 %以上的活力。 展开更多
关键词 重组酯酶 基因表达 酶学性质 ralstonia eutropha CH34 大肠杆菌 酯酶活性
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利用GFP标记的Ralstonia solanacearum鉴定马铃薯青枯病抗性 被引量:3
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作者 陈卓 肖熙鸥 +3 位作者 陈曙 李可 邹华芬 金辉 《中国瓜菜》 CAS 北大核心 2021年第1期35-41,共7页
青枯病是危害马铃薯产业的重要病害,选育与推广抗青枯病品种是防控青枯病最高效的手段。目前已有的人工接种和田间抗性鉴定的方法不能有效区分处于潜伏侵染与完全抵抗侵染的材料,而利用GFP标记的Ralstonia solanacearum能够有效解决上... 青枯病是危害马铃薯产业的重要病害,选育与推广抗青枯病品种是防控青枯病最高效的手段。目前已有的人工接种和田间抗性鉴定的方法不能有效区分处于潜伏侵染与完全抵抗侵染的材料,而利用GFP标记的Ralstonia solanacearum能够有效解决上述问题。笔者通过电击转化法将广宿主载体pBBR1MCS2-Tac-EGFP导入青枯菌P2中,成功获得了能够稳定遗传且不影响其致病力并带有绿色荧光报告的青枯菌菌株P2-Tac-EGFP。通过对感抗材料进行接种,结果表明,P2-Tac-EGFP能够有效区分感抗材料,并且P2-Tac-EGFP在感病材料的根部和茎部大量分布,而在抗病材料的根部仅有少量分布。综上所述,利用GFP标记的R.solanacearum能够快速准确地鉴定马铃薯青枯病抗性。 展开更多
关键词 马铃薯 ralstonia solanacearum GFP 抗性
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繁殖体数量对外来Ralstonia solanacearum在红壤中入侵潜力的影响 被引量:1
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作者 马超 庄睿花 +6 位作者 王时聪 许邶 柴如山 屠人凤 朱林 李道林 郜红建 《安徽农业大学学报》 CAS CSCD 2019年第4期665-670,共6页
外来种的繁殖体压力因可调控其他入侵影响因素(如外来种特性)而备受入侵生态学家的关注。前人研究指出剂量-响应曲线能够定量分析繁殖体数量对入侵潜力的影响,但关于该曲线形状的认识尚未统一。利用土壤微宇宙,比较了初始接种量为10^3(P... 外来种的繁殖体压力因可调控其他入侵影响因素(如外来种特性)而备受入侵生态学家的关注。前人研究指出剂量-响应曲线能够定量分析繁殖体数量对入侵潜力的影响,但关于该曲线形状的认识尚未统一。利用土壤微宇宙,比较了初始接种量为10^3(PP3)、10^5(PP5)、10^7(PP7)和10^9 CFU·g^-1(PP9)的青枯病菌Ralstonia solanacearum进入土壤3和42 d后的存活量,并对初始接种量和存活量之间关系进行拟合,试图探清地下部微生物入侵的剂量-响应曲线状况。研究发现,不同接种量处理外来R.solanacearum接入土壤3 d后的存活量差异显著(P<0.05),从大到小依次为PP9、PP7、PP5和PP3,初始接种量和存活量之间拟合的剂量-响应曲线为指数型;而外来Ralstonia solanacearum接入土壤42 d后,除了初始接种量为109 CFU·g^-1处理的存活量稍高外,其余处理间差异不显著,此时的剂量-响应曲线呈直线型(斜率约为0)。结果表明,外来病原菌入侵土壤时的剂量-响应关系会随其进入土壤时间的延长而改变,前期为指数型,后期转为直线型,这说明外来R.solanacearum在红壤中的入侵潜力在入侵前期随繁殖体数量增加而呈指数增长,在入侵后期不受繁殖体数量的影响。 展开更多
关键词 微生物入侵 繁殖体数量 剂量-响应曲线 青枯菌 红壤
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Silicon impacts on soil microflora under Ralstonia Solanacearum inoculation 被引量:4
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作者 LIN Wei-peng JIANG Ni-hao +4 位作者 PENG Li FAN Xue-ying GAO Yang WANG Guo-ping CAI Kun-zheng 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2020年第1期251-264,共14页
Silicon(Si) can increase plant resistance against bacterial wilt caused by Ralstonia solanacearum and enhance plant immune response. However, whether Si alleviates soil-borne disease stress through altering soil micro... Silicon(Si) can increase plant resistance against bacterial wilt caused by Ralstonia solanacearum and enhance plant immune response. However, whether Si alleviates soil-borne disease stress through altering soil microbial community component and diversity is not clear. In this study, effects of Si application under R. solanacearum inoculation with or without plant on soil bacterial and fungal communities were investigated through high-throughput pyrosequencing technique. The results showed that Si addition significantly reduced bacterial wilt incidence. However, Si did not reduce the amount of R. solanacearum in rhizosphere soil. Principal components analysis showed that soil microbial community composition was strongly influenced by Si addition. Total 63.7% bacterial operational taxonomic units(OTUs) and 43.8% fungal OTUs were regulated by Si addition regardless of the presence of tomato plants, indicating the independent effects of Si on soil microbial community. Si-added soil harbored a lower abundance of Fusarium, Pseudomonas, and Faecalibacterium. Our finding further demonstrated that exogenous Si could significantly influence soil microbial community component, and this may provide additional insight into the mechanism of Si-enhanced plant resistance against soil-borne pathogens. 展开更多
关键词 bacterial wilt deep pyrosequencing ralstonia solanacearum SILICON soil microbial community
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Physiological response and phenolic metabolism in tomato (Solanum lycopersicum) mediated by silicon under Ralstonia solanacearum infection 被引量:3
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作者 FAN Xue-ying LIN Wei-peng +2 位作者 LIU Rui JIANG Ni-hao CAI Kun-zheng 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2018年第10期2160-2171,共12页
Bacterial wilt, caused by Ralstonia solanacearum(Rs) is a serious soil-borne disease and silicon can enhance tomato resistance against this disease. However, few studies have focused on the mechanisms of Si-mediated... Bacterial wilt, caused by Ralstonia solanacearum(Rs) is a serious soil-borne disease and silicon can enhance tomato resistance against this disease. However, few studies have focused on the mechanisms of Si-mediated pathogen resistance from the rhizosphere perspective. In this study, two tomato genotypes, HYT(susceptible) and H7996(resistant), were used to investigate the effects of silicon application on disease inhibition, root growth, and organic acid content in both roots and root exudates under R. solanacearum infection. The results showed that Si application significantly suppressed bacterial wilt in HYT, but had no effect in H7996. Silicon concentrations in roots, stems and leaves of tomato were significantly increased by Si treatment under R. solanacearum inoculation. In HYT, Si application increased root dry weight by 22.8-51.6% and leaf photosynthesis by 30.6-208.0%, and reduced the concentrations of citric acid in root exudates by 71.4% and in roots by 83.5%. However, organic acids did not influence R. solanacearum growth. Results also demonstrated that salicylic acid(SA) content in roots was significantly increased by silicon addition for H7996 and exogenous SA application could reduce bacterial wilt disease index. Collectively, these results suggest that Si-modulated phenolic compound metabolism in roots or root exudates, especially citric acid and SA, may be a potential mechanism in the amelioration of bacterial wilt disease by Si. 展开更多
关键词 SILICON ROOT root exudates Solanum lycopersicum ralstonia solanacearum organic acids
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Effect of K1, K2 Anti-Bacterial Agents on Tobacco Ralstonia Solanacearum 被引量:8
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作者 Anan Wang Zhifeng Zhao +1 位作者 Zhenzhen Liu Yixin Liu 《Engineering(科研)》 2010年第11期930-934,共5页
The tobacco Ralstonia Solanacearum were both cultured on nutrient agar plates and inoculated in seedling stage of tobacco, then treated with K1 and K2, two anti-bacterial agents, at a serial con-centrations to study t... The tobacco Ralstonia Solanacearum were both cultured on nutrient agar plates and inoculated in seedling stage of tobacco, then treated with K1 and K2, two anti-bacterial agents, at a serial con-centrations to study their inhibitory efficiency. The result indicated that K1 can inhibit R. Solanacearum growth entirely, at the concentration range from 1/50 to 1/5000. K2 can reach the same result at the concentration range from 1/50 to 1/50000. Compared with the control plates, K1, at the concentration 1/50000, had no significant differences, and the average number of colony per plate was 112-115. The immature tobacco shown wilt as soon as inoculated with R. Solanacearum, and recovered gradually after using K1, K2. The densities of microbial suspension, handled by K1, K2 within 10 hs, were both significantly lower than the controlled ones. The optical microscopy also shown that handled microbial body differed from the controlled, whose body was regular short, rod shape as opposed to the handled ones with irregular rod shape and damaged body. All the results indicated that K1 and K2 both had inhibitory effects on tobacco R. Solanacearum, and K2 was more efficient than K1. 展开更多
关键词 ANTI-BACTERIAL AGENTS K1 K2 Inhibition TOBACCO ralstonia SOLANACEARUM
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Ralstonia eutropha CH34酯酶基因的克隆和序列分析 被引量:2
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作者 宋水山 Alexander Steinbchel 《微生物学报》 CAS CSCD 北大核心 2001年第4期402-407,共6页
采用含α 乙酸萘酯和固兰RR的表面琼脂法从RalstoniaeutrophaCH34的基因文库中筛选酯酶基因estA ,对含有estA的 1 7kbDNA片段的核甘酸序列分析表明 ,该基因全长82 5bp,编码由 2 75个氨基酸组成的EstA蛋白 ,分子量为 30 785D。经推导氨... 采用含α 乙酸萘酯和固兰RR的表面琼脂法从RalstoniaeutrophaCH34的基因文库中筛选酯酶基因estA ,对含有estA的 1 7kbDNA片段的核甘酸序列分析表明 ,该基因全长82 5bp,编码由 2 75个氨基酸组成的EstA蛋白 ,分子量为 30 785D。经推导氨基酸序列的同源性分析 ,发现EstA与参与芳香化合物代谢中间位裂解途径的水解酶有很高的同源性。 展开更多
关键词 ReutrophaCH34 酯酶基因 DNA序列 大肠杆菌 克隆 表达
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Detection and Analysis of Number of Ralstonia solanacearum in Soil before Winter Tillage in Fuzhou Tobacco-growing Area, Jiangxi Province 被引量:1
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作者 Zhang Zhigao Hu Liwei +3 位作者 Feng Xiaohu Mou Wenjun Wang Libing Zhang Rui 《Plant Diseases and Pests》 CAS 2019年第5期1-3,10,共4页
[Objective]The paper was to optimize the tobacco planting area in Fuzhou City and to prevent the outbreak of tobacco bacterial wilt in large area.[Method]At the end of 2017,soil samples were collected from plots plann... [Objective]The paper was to optimize the tobacco planting area in Fuzhou City and to prevent the outbreak of tobacco bacterial wilt in large area.[Method]At the end of 2017,soil samples were collected from plots planned to be planted with tobacco in the following year in Yihuang,Guangchang,Lichuan and Le’an counties.[Result]Among 352 plots,116 plots were infected by Ralstonia solanacearum,while 236 plots were free of the pathogen,and the infected plots accounted for 32.95% of total plots.Among them,75 plots exceeded the order of magnitudes of 103,accounting for 21.31% of total plots and 64.66% of infected plots.It is suggested that the plots with an order of magnitude above 103 should be pretreated with quicklime or purple soil,or conducted crop rotation,or seeds must be directly abandoned;the dosage of biocontrol agents should be increased in planting.The plots with an order of magnitude below 103 should be pretreated with quicklime or purple soil,and the dosage of biocontrol agents should be increased in planting.[Conclusion]The results provide reliable theoretical basis and data support for soil improvement and bacterial wilt control. 展开更多
关键词 TOBACCO ralstonia SOLANACEARUM Winter TILLAGE BIOCONTROL
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pBBR1MCS2-Tac-EGFP广宿主载体适宜标记Ralstonia solanacearum
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作者 肖熙鸥 林文秋 +3 位作者 陈卓 邹春香 金辉 邹华芬 《热带作物学报》 CSCD 北大核心 2021年第6期1700-1705,共6页
利用标记基因追踪病原菌在植物体内的入侵和定殖,是研究病原菌-寄主互作的重要手段。本研究利用电转化法将广宿主载体pBBR1MCS2-Tac-EGFP导入青枯雷尔氏菌(Ralstonia solanacearum)GMI1000菌株中,获得了青枯雷尔氏菌带绿色荧光标记的转... 利用标记基因追踪病原菌在植物体内的入侵和定殖,是研究病原菌-寄主互作的重要手段。本研究利用电转化法将广宿主载体pBBR1MCS2-Tac-EGFP导入青枯雷尔氏菌(Ralstonia solanacearum)GMI1000菌株中,获得了青枯雷尔氏菌带绿色荧光标记的转化子。转接试验结果表明,转化子的抗生素抗性和绿色荧光强度有良好的遗传稳定性。pBBR1MCS2-Tac-EGFP不影响GMI1000菌株的致病力,且EGFP蛋白能够在植物中稳定表达。灌根法接种试验结果表明,病原菌在第1天即完成对根系的侵染,并在第6天扩散至其他组织,随后造成植株萎蔫。研究结果表明所获转化子可用于后续的病原菌侵染机理等方面的研究。 展开更多
关键词 青枯雷尔氏菌 绿色荧光标记 侵染动态 pBBR1MCS2-Tac-EGFP载体
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Differential Space-Time Expression of StLTPbl Gene Between Resistant and Susceptible Potato Genotypes in Response to Ralstonia solanacearum
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作者 GAO Gang JIN Li-ping +1 位作者 XIE Kai-yun QU Dong-yu 《Agricultural Sciences in China》 CAS CSCD 2008年第6期713-725,共13页
This study is to investigate the role of lipid transfer protein (LTP1) gene of potato (Solanum tuberosum) in bacterial wilt (Ralstonia solanacearum) resistance. A novel cDNA clone encoding nsLTP was isolated fro... This study is to investigate the role of lipid transfer protein (LTP1) gene of potato (Solanum tuberosum) in bacterial wilt (Ralstonia solanacearum) resistance. A novel cDNA clone encoding nsLTP was isolated from cultivated potato (Solanum tuberosum) infected with R. solanacearum by 5'-rapid amplification of cDNA ends (RACE). The temporal and spatial expression of StLTPbl was studied during the early stages of potato-R, solanacearum interaction by reverse transcriptase PCR (RT-PCR) and Northern blotting. The sequence analysis of the cloned cDNA, named StLTPbl, showed 691 bp which encoded a type 1 nsLTP of 91 amino acids. Construction of a phylogenic tree showed that StLTPbl is well conserved in the coding region with high identity at the amino acid level with other Solanaceae nsLTPs. The temporal and spatial expression of StLTPbl was studied during the early stages of potato-R, solanacearum interaction. StLTPbl transcription is induced faster and transcripts accumulate to higher concentrations in resistant compared with susceptible genotypes by the pathogen. Dominant differences in the pathogen-induced gene expression pattern between the upper and lower leaves and stems were observed within the same genotypes. In situ hybridization results showed that the StLTPbl mRNA was localized in phloem cells of vascular tissues in potato leaf and stem tissues after pathogen infection. Salicylic acid, methyl jasmonate and abscisic acid could induce StLTPbl gene expression without significant difference between the upper and lower tissues. These abiotic elicitors could produce a long-lastingeffect on the StLTPbl during early stages of potato-R, solanacearum interaction. Differential expression of StLTPbl gene between resistance and susceptible potato genotypes in response to R. solanacearum suggests that this gene plays a key role in plant defense mechanisms. 展开更多
关键词 Solanum tuberosum NSLTP gene expression ralstonia solanacearum
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