Institute of Plant Protection, Key Lab of Food Quality and Safety of Jiangsu Province-State, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, P.R. China.
Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, P.R. China.
Plant Dis. 2022 Jan;106(1):231-246. doi: 10.1094/PDIS-06-21-1223-RE. Epub 2022 Jan 21.
Yields of edible rhizome from cultivation of the perennial hydrophyte lotus () can be severely reduced by rhizome rot disease caused by species. There is a lack of rapid field-applicable methods for detection of these pathogens on lotus plants displaying symptoms of rhizome rot. (91%) and (9%) were identified at different frequencies from lotus samples showing symptoms of rhizome rot. Because these two species can cause different severity of disease and their morphology is similar, molecular diagnostic-based methods to detect these two species were developed. Based on the comparison of the mitochondrial genome of the two species, three specific DNA loci targets were found. The designed primer sets for conventional PCR, quantitative PCR, and loop-mediated isothermal amplification (LAMP) precisely distinguished the above two species when isolated from lotus and other plants. The LAMP detection limits were 10 pg/μl and 1 pg/μl of total DNA for and , respectively. We also carried out field-mimicked experiments on lotus seedlings and rhizomes (including inoculated samples and field-diseased samples), and the results indicated that the LAMP primer sets and the supporting portable methods are suitable for rapid diagnosis of the lotus disease in the field. The LAMP-based detection method will aid in the rapid identification of whether or is infecting lotus plants with symptoms of rhizome rot and can facilitate efficient pesticide use and prevent disease spread through vegetative propagation of -infected lotus rhizomes.
由 引起的根茎腐烂病会严重降低多年生水生植物莲藕()的可食用根茎产量。对于表现出根茎腐烂症状的莲藕植物，缺乏快速适用于现场的检测这些病原体的方法。从表现出根茎腐烂症状的莲藕样本中，不同频率地鉴定出 （91%）和 （9%）。由于这两个物种可以引起不同严重程度的疾病，并且它们的形态相似，因此开发了基于分子诊断的方法来检测这两个物种。基于两个物种的线粒体基因组的比较，发现了三个特定的 DNA 靶标。设计的用于常规 PCR、定量 PCR 和环介导等温扩增 (LAMP) 的引物组能够从莲藕和其他植物中准确地区分这两个物种。LAMP 的检测限分别为 和 的 10 pg/μl 和 1 pg/μl 的总 DNA。我们还在莲藕幼苗和根茎（包括接种样本和田间患病样本）上进行了现场模拟实验，结果表明 LAMP 引物组和支持的便携式方法适用于现场快速诊断莲藕病害。基于 LAMP 的检测方法将有助于快速识别是否有 或 感染有根茎腐烂症状的莲藕植物，并有助于通过受感染的莲藕根茎的营养繁殖来有效使用农药和防止疾病传播。
