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Characterization of a new curtovirus, Pepper yellow dwarf virus, from chile pepper in New Mexico.

机译:从新墨西哥州的辣椒中鉴定出一种新的轮状病毒,辣椒黄矮病毒。

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摘要

Curly top disease in chile pepper caused by curtoviruses is a challenging disease in New Mexico. Currently, curtoviruses in New Mexico have not been studied thoroughly. No one approach manages curly top disease in chile efficiently. So, the objectives of this project are to characterize curtoviruses in New Mexico, and identify curtovirus-resistant lines in chile for control of curly top disease. This project includes three parts: (1) Identifying curtovirus strains in New Mexico; (2) Antibody production for curtovirus dectection by ELISA; (3) Screening for curtovirus resistant chile plants.;Using eight primers designed for PCR, complete genomes of three curtoviruses isolated from chile pepper samples representing the three groups of curtoviruses in New Mexico were sequenced. Comparisons of whole sequences of the genomes revealed that DGSW171601 isolate belongs to Beet mild curly top virus (BMCTV). LRME27601 isolate belongs to Beet severe curly top virus (BSCTV), and LJN17601 isolate presents a new curtovirus species designated as Pepper yellow dwarf virus (PeYDV). Pepper yellow dwarf virus has a circular single stranded 2959 by DNA genome. This new curtovirus species may be the result of recombination between BMCTV-W4 and BSCTV and mutations in the genome, especially in the region of ORFs C1 to C4.;The coat protein gene was cloned from the new curtovirus, Pepper yellow dwarf virus (PeYDV) and inserted into the pRSET-A vector to express the coat protein in E. coli. Polyclonal antibodies against curtoviruses were produced in rabbits using the coat protein as antigen. These polyclonal antibodies were purified through the DEAE column. Testing their immune function showed that these polyclonal antibodies can be used directly for ELISA testing to detect the presence of curtoviruses in chile plants.;NM64, Tabasco, and NuMex chile varieties were inoculated with Agrobacterium carrying a recombinant curtovirus in Ti-plasmid for virus resistant screening. ELISA and PCR were used to detect the presence of the curtovirus in inoculated plants. The data showed that NM64 chile variety is susceptible to curtoviruses. The infection rate was approximately 68%. Tabasco and NuMex chile varieties are partially resistant to curtoviruses. The infection rates were approximately 16% and 28%, respectively.
机译:在新墨西哥州,由轮状病毒引起的辣椒的卷毛病是具有挑战性的疾病。目前,尚未对新墨西哥州的弯曲病毒进行深入研究。没有一种方法可以有效地管理智利的卷发病。因此,该项目的目标是表征新墨西哥州的弯曲病毒,并确定辣椒中抗弯曲病毒的品系,以控制卷曲顶病。该项目包括三个部分:(1)确定新墨西哥州的弧菌病毒株; (2)通过ELISA检测弯曲病毒的抗体生产; (3)筛选耐弯曲病毒的智利植物。;使用设计用于PCR的八种引物,对代表新墨西哥州三组弯曲病毒的辣椒样品分离的三种弯曲病毒的完整基因组进行测序。基因组全序列的比较表明,DGSW171601分离株属于甜菜轻度卷曲病毒(BMCTV)。 LRME27601分离株属于甜菜严重卷曲顶病毒(BSCTV),LJN17601分离株具有一种新的弯曲病毒种,称为胡椒黄矮病毒(PeYDV)。辣椒黄矮病毒的DNA基因组具有环状单链2959。这种新的弯曲病毒可能是BMCTV-W4和BSCTV重组以及基因组突变的结果,特别是在ORF C1至C4区域。;外壳蛋白基因是从新的弯曲病毒辣椒黄矮病毒(PeYDV)克隆而来的)插入pRSET-A载体以在大肠杆菌中表达外壳蛋白。使用外壳蛋白作为抗原在兔中产生了针对弯曲病毒的多克隆抗体。这些多克隆抗体通过DEAE柱纯化。测试其免疫功能表明,这些多克隆抗体可直接用于ELISA测试,以检测辣椒植物中弯曲病毒的存在。;将农杆菌接种于携带Ti-质粒的重组弯曲杆菌病毒的农杆菌农杆菌NM64,塔巴斯科州和NuMex智利品种中以抵抗病毒筛选。 ELISA和PCR用于检测接种植物中弯曲病毒的存在。数据表明,NM64辣椒品种易受轮状病毒感染。感染率约为68%。塔巴斯科州(Tabasco)和NuMex智利品种对弯曲病毒有​​部分抗性。感染率分别约为16%和28%。

著录项

  • 作者

    Lam, Nhan D.;

  • 作者单位

    New Mexico State University.;

  • 授予单位 New Mexico State University.;
  • 学科 Biology Molecular.;Agriculture Plant Pathology.;Biology Virology.
  • 学位 Ph.D.
  • 年度 2009
  • 页码 77 p.
  • 总页数 77
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分子遗传学;植物病理学;
  • 关键词

  • 入库时间 2022-08-17 11:37:41

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