INTERACTION OF TURNIP CRINKLE VIRUS (TCV) AND ITS HOST PLANTS

芜菁皱缩病毒 (TCV) 与其寄主植物的相互作用

• 批准号: 7627616
• 负责人: PAUL TWIGG
• 金额: $ 5.36万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7627616
• 关键词: Acetylcysteine; Alkaloids; Amino Acids; Anabolism; Arabidopsis; Behavior; Binding; Bone callus; Capsid Proteins; Chamaecrista; Collaborations; Complementary DNA; Computer Retrieval of Information on Scientific Projects Database; Consumption; Dental crowns; Deposition; Disease; Eye; Fabaceae; Family; Funding; Future; Genbank; Genes; Genetic; Genome; Genomics; Glean; Grant; Harvest; Individual; Institution; Left; Leghemoglobin; Libraries; Lignin; Lilium; Maize; Manuscripts; Metabolism; Microarray Analysis; Mutate; Names; Nitrogen; Nodule; Open Reading Frames; Oxygen; Panicum; Patient currently pregnant; Plant Leaves; Plant Model; Plant Roots; Plants; Poaceae; Polymerase Chain Reaction; Population; Production; Proteins; Publications; Publishing; RNA; RNA Interference; Recombinant Proteins; Research; Research Personnel; Research Project Grants; Resources; Rhizobium; Seedling; Sequence Analysis; Source; Structure; Students; Symbiosis; Time; Tissues; Transcript; Transcription Coactivator; Turnip - dietary; United States National Institutes of Health; Veratrum; Virus; Work; Zea mays; base; cDNA Arrays; cDNA Library; cyclopamine; expression vector; genetic analysis; infancy; interest; mutant; stem; toe corn; transcription factor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. My main research project is focused on the interaction of Turnip Crinkle Virus (TCV) and its host plants. We are analyzing the genetic interactions of TCV and the model plant Arabidopsis. Currently, we are using two strains of TCV, a wild type strain and a strain developed by the Morris lab at UNL with one amino acid altered in the viral coat protein called R6a. Wild type TCV coat protein shows a binding with a transcriptional activator in Arabidopsis belonging to the NAC family of transcription factors. This interaction causes RNA silencing in the infected plants. The mutated R6a TCV does not show this interaction and causes a more severe and symptomatically different disease. We are analyzing this interaction by inoculating wild type Arabidopsis with normal and mutant TCV individually, as well as a mock inoculation with no virus. We then harvested leaves after 24 and 48 hrs form all three plant populations. We have extracted total RNA from all six tissues in triplicate. These RNAs will soon be used in microarray analysis of Arabidopsis whole genome cDNA arrays. Transcripts identified as being the most consistently up or down regulated from these arrays will be further studied in the near future using real-time PCR to verify the microarray results. I also have several other projects underway mostly in collaboration with Dr. Gautam Sarath, but also with Dr. Christian Tobias and Dr. John Vogel all of the USDA. With Drs. Sarath and Tobias, I have been working on a genomics inititative characterizing the genetics of the biofuel plant Switchgrass (Panicum virgatum). My students and I have to date constructed cDNA libraries from Switchgrass callus tissue, stems, leaves, crowns, and seedlings. Together with Dr. Tobias, we have sequenced and analyzed the 5' end of 15,272 cDNA clones from the leaf, callus, stem ,and crown libraries. All of these sequences are soon to be deposited in GenBank bearing my and my students names as well as the appropriate INBRe acknowledgement. From these sequences, we have gleaned many genes important in a variety of metabolic processes for further characterization. Chief amongst these are the genes involved in lignin biosynthesis which will soon be the basis of our first publications on this material. I am currently beginning a similar genomics initiative with Dr. John Vogel studying another biofuel grass called Bradypodium distachyon. This project is in its infancy, but we are currently constructing four cDNA libraries from various tissues from this plant. I am also working with Dr. Sarath on a genomics study of a prairie legume called Partridge Pea (Chamaecrista fasciculata). This plant is an important prairie species that participates in a symbiotic nitrogen-fixing symbiosis with Rhizobium forming root nodules. We have constructed a cDNA library from the nodule tissue of this plant, and have begun to characterize the 5' end of various sequences. We have isolated a clone for a symbiotic protein called leghemoglobin which we have selected for further characterization. We are currently concentrating on subcloning the open-reading frame into an appropriate expression vector for the production of recombinant protein. We intend to analyze the oxygen-binding behavior and structure of the leghemoglobin protein. This work will shortly produce its first manuscripts. My last project involves a plant called corn lily (Veratrum californicum). This plant is of interest due to its production of the teratogenic and carcinogenic steroidal alkaloid called cyclopamine. This compound is largely concentrated in the roots of the corn lily, but consumption of its leaves by pregnant ewes can produce somewhat gruesome offspring with only one eye or both eyes merged into one socket. To study this interesting plant, we have constructed cDNA libraries from the roots and leaves. We have also analyzed the differences in the transcriptomes of these tissues using cDNA-AFLP analysis. We have isolated and sequenced in excess of 100 individual AFLP fragments which are unique to one tissue or the other. We are currently analyzing these sequences with the hope of identifying some of the genetic factors contributing to cyclopamine synthesis. We hope to publish this work shortly.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 我的主要研究项目的重点是萝卜皱纹病毒（TCV）及其宿主植物的相互作用。 我们正在分析TCV和模型植物拟南芥的遗传相互作用。 目前，我们使用了两种TCV菌株，一种野生型应变和由莫里斯实验室在UNL处开发的菌株，其中一种称为R6A的病毒涂层蛋白中改变的氨基酸。 野生型TCV涂层蛋白在属于NAC转录因子家族的拟南芥中与转录激活剂的结合。 这种相互作用会导致感染植物中的RNA沉默。 突变的R6A TCV并未显示这种相互作用，并引起更严重和症状不同的疾病。 我们正在通过单独接种具有正常和突变体TCV的野生型拟南芥来分析这种相互作用，以及无病毒的模拟接种。 然后，我们在24小时和48小时后收获叶子，这三个植物种群。 我们一式三份从所有六个组织中提取了总RNA。 这些RNA很快将用于拟南芥整个基因组cDNA阵列的微阵列分析。 使用实时PCR验证微阵列结果，将在不久的将来进一步研究被确定为从这些阵列中调节的最稳定的转录本。 我还与Gautam Sarath博士合作，还与Christian Tobias博士和John Vogel博士一起进行了其他几个项目。 与博士。萨拉斯（Sarath）和托比亚斯（Tobias），我一直在研究一种基因组学的发起，该起始特征着生物燃料植物开关草（Panicum virgatum）的遗传学。 我和我的学生必须通过开关草，茎，树叶，冠和幼苗的callus愈伤组织构造的cDNA库。 与Tobias博士一起，我们从叶子，愈伤组织，茎和皇冠库中测序并分析了15,272个cDNA克隆的5'末端。 所有这些序列很快就会存放在带有我和我的学生名字的GenBank中，以及适当的inbre确认。 从这些序列中，我们收集了许多在多种代谢过程中重要的基因，以进一步表征。 其中的主要是木质素生物合成涉及的基因，这些基因很快将成为我们对该材料的第一本出版物的基础。 我目前正在开始使用类似的基因组学计划，约翰·沃格尔（John Vogel）博士研究了另一种名为Bradypodium distachyon的生物燃料草。 该项目还处于起步阶段，但我们目前正在该工厂的各种组织中构建四个cDNA文库。 我还与Sarath博士一起研究了一项名为Partridge Pea（Chamaecrista fasciculata）的草原豆科植物的基因组学研究。 该植物是一种重要的草原物种，参与共生氮的共生，形成根结节。 我们已经从该植物的结节组织构建了一个cDNA库，并开始表征各种序列的5'端。 我们已经分离了一种称为leghemoglobin的共生蛋白的克隆，我们选择了以进一步的表征。 我们目前正在集中于将开读框架亚克隆到适当的表达载体中，以生产重组蛋白。 我们打算分析leghemoglobin蛋白的氧结合行为和结构。 这项工作将很快产生其第一批手稿。 我的最后一个项目涉及一种称为玉米莉莉（Veratrum californicum）的植物。 该植物引起了人们的关注，这是由于其生产称为环孢胺的致癌性和致癌类固醇生物碱。 这种化合物主要集中在玉米百合的根部，但是怀孕的母羊消耗其叶子可以产生一些可怕的后代，只有一只眼睛或两只眼睛合并为一个插座。 为了研究这种有趣的植物，我们已经从根和叶子中构建了cDNA文库。 我们还使用cDNA-AFLP分析分析了这些组织的转录组的差异。 我们已经分离并测序了100个单独的AFLP片段，这些片段是一家组织或另一个组织所独有的。 我们目前正在分析这些序列，以期识别一些造成环保胺合成的遗传因素。 我们希望很快发布这项工作。