Hamim Islam, Suzuki Jon Y, Borth Wayne B, Melzer Michael J, Wall Marisa M, Hu John S
Department of Plant Pathology, Bangladesh Agricultural University, Mymensingh, Bangladesh.
USDA-ARS, Daniel K. Inouye U.S. Pacific Basin Agricultural Research Center, Hilo, HI, United States.
Front Microbiol. 2022 Aug 25;13:930329. doi: 10.3389/fmicb.2022.930329. eCollection 2022.
Viral diseases in plants have a significant impact on agricultural productivity. Effective detection is needed to facilitate accurate diagnosis and characterization of virus infections essential for crop protection and disease management. For sensitive polymerase chain reaction (PCR)-based methods, it is important to preserve the integrity of nucleic acids in plant tissue samples. This is especially critical when samples are collected from isolated areas, regions distant from a laboratory, or in developing countries that lack appropriate facilities or equipment for diagnostic analyses. RNA provides effective, reliable sample storage by stabilizing both RNA and DNA in plant tissue samples. Our work indicated that total RNA or DNA extracted from virus-infected leaf samples preserved in RNA was suitable for reverse transcription polymerase chain reaction (RT-PCR), PCR, Sanger sequencing, high-throughput sequencing (HTS), and enzyme-linked immunosorbent assay (ELISA)-based diagnostic analyses. We demonstrated the effectiveness of this technology using leaf tissue samples from plants with virus symptoms grown in farmers' fields in Bangladesh. The results revealed that RNA technology was effective for detection and characterization of viruses from samples collected from remote areas and stored for extended periods. Adoption of this technology by developing countries with limited laboratory facilities could greatly increase their capacity to detect and diagnose viral infections in crop plants using modern analytical techniques.
植物病毒病对农业生产力有重大影响。需要进行有效的检测，以促进对病毒感染的准确诊断和特征描述，这对于作物保护和病害管理至关重要。对于基于敏感聚合酶链反应（PCR）的方法，保持植物组织样本中核酸的完整性很重要。当样本从偏远地区、远离实验室的区域或缺乏适当诊断分析设施或设备的发展中国家采集时，这一点尤为关键。RNA 通过稳定植物组织样本中的RNA和DNA，提供了有效、可靠的样本储存方法。我们的研究表明，保存在RNA中的病毒感染叶片样本中提取的总RNA或DNA适用于逆转录聚合酶链反应（RT-PCR）、PCR、桑格测序、高通量测序（HTS）以及基于酶联免疫吸附测定（ELISA）的诊断分析。我们利用孟加拉国农民田间出现病毒症状的植物叶片组织样本证明了该技术的有效性。结果表明，RNA技术对于从偏远地区采集并长期保存的样本中的病毒检测和特征描述是有效的。实验室设施有限的发展中国家采用这项技术，可以极大地提高其使用现代分析技术检测和诊断作物植物病毒感染的能力。
