樟疫霉高信赖度特异性检测靶标PHYCI_587572的鉴定及基于该靶标的试纸条技术研究
摘 要
疫霉属（Phytophthora）是属于卵菌门的一类病原菌，其寄主范围大、发病重且较难控制，能对植物产量造成巨大损失。林木上疫霉菌的分类与鉴定是目前病原生物中分类、鉴定中最困难的属之一。疫霉菌引起的植物病害在症状上与一些其他病原菌引起的病害非常相似，而且疫霉菌的分离和培养都较困难。樟疫霉（Phytophthora cinnamomi）引起的病害是林木植物的毁灭性病害之一。目前樟疫霉的检验检疫主要依赖诱捕、分离、病原菌的形态观察等传统检测方法，耗时较长且效率低，未能满足快速、准确、灵敏的检测需求，因此快速分子检测樟疫霉，阻断传播源头是当前阻止病害传播的重要途径之一。无论是何种检测技术，其可靠性取决于特异性分子靶标的发掘。前人发掘的分子检测靶标，由于缺乏基因组学的支持，没有经过系统筛选，检测结果往往可信度不够。因此挖掘在种间具有差异性，但在种内菌株间具有较高保守性的基因片段作为分子检测靶标是挖掘分子检测靶标的关键。为此，本研究采用生物信息学和比较基因组学方法筛选并鉴定出新特异性靶标，并基于新发掘靶标采用重组酶聚合酶扩增技术（Recombinase Polymerase Amplification，RPA）和侧流层析技术（Lateral Flow Dipstick, LFD）建立樟疫霉（P. cinnamomi）的试纸条快速检测体系。
关键词：樟疫霉；PHYCI_587572；植物检疫；RPA技术；LFD检测
PHYCI_587572: an RxLR Effector Gene and New Biomarker in A Recombinase Polymerase Amplification Assay for Rapid Detection of Phytophthora cinnamomi
ABSTRACT
The genus Phytophthora consists of many species that cause important diseases in ornamental, agronomic, and forest ecosystems worldwide. Phytophthora spp. are responsible for serious disease worldwide and can occur on a wide range of different crops which cause a great loss. Phytophthora cinnamomi is a devastating pathogen causing root and crown rot and dieback diseases of nearly 5000 plant species. Accurate and Rapid detection of P. cinnamomi plays a fundamental role within the current disease prevention and management programs. With the advent of molecular technology, other faster and more cost-effective techniques including Polymerase Chain Reaction (PCR), real-time fluorescence quantitative PCR, isothermal diagnostic assays having become the mainstream in rapid detection of many Phytophthora species. The reliability of any detection technique depends on the discovery of specific molecular detection targets. Due to the lack of genomics support, molecular detection targets discovered by previous researchers have not been systematically screened, which often leads to problems in the credibility of detection results. Thus, an ideal detection assay must be sensitive enough to detect low levels of P. hibernalis in both symptomatic and asymptomatic plants. Additionally, it must be specific to P. cinnamomi , as misidentification of relatively low-risk Phytophthora species may lead to unnecessary management and removal of plants, which can be costly..Therefore, A basic RPA (Recombinase Polymerase Amplification) assay was established for P. cinnamomi and a rapid assay including 25-min recombinase polymerase amplification and 5-min visualization using lateral flow dipsticks was developed for detecting P. cinnamomi. Meanwhile, based on bioinformatics and comparative genomics methods, some new special molecular detection targets for P. cinnamomi were also identified from genomic
sequence data.
Key words: Phytophthora cinnamomi; Target; Disease diagnosis; Recombinase Polymerase Amplification; Lateral Flow Dipstick
以上是毕业论文文献综述，课题毕业论文、任务书、外文翻译、程序设计、图纸设计等资料可联系客服协助查找。
