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Rapid Detection of Wheat Blast Pathogen magnaporthe oryzae triticum Pathotype Using Genome-Specific Primers and Cas12a-mediated Technology

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Engineering 2021年第9期7卷 1326-1335页

作者： Houxiang Kang Ye Peng Kangyu Hua Yufei Deng Maria Bellizzi Dipali Rani Gupta Nur Uddin Mahmud Alfredo S.Urashima Sanjoy Kumar Paul Gary Peterson Yilin Zhou Xueping Zhou Md Tofazzal Islam Guo-Liang Wang State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193China Department of Plant Pathology The Ohio State UniversityColumbusOH 43210USA Institute of Biotechnology and Genetic Engineering(IBGE) Bangabandhu Sheikh Mujibur Rahman Agricultural UniversityDhaka 1706Bangladesh Centro de Ciências Agrárias Universidade Federal de São CarlosArarasSP CEP 13600-000Brazil Foreign Disease-Weed Science Research Unit(FDWSRU) United States Department of Agriculture-Agricultural Research Service(USDA-ARS)Ft.DetrickMD 21702USA

Wheat blast,caused by the fungus magnaporthe oryzae triticum(MoT)pathotype,is a devastating disease persistent in South America and *** MoT generally fails to cause visual symptoms in wheat until the heading stage when the infection would have advanced,disease control by fungicide application solely based on the detection of visual symptoms is *** develop an accurate and sensitive method to detect MoT at the seedling and vegetative stages for disease control,we sequenced the genomes of two MoT isolates from Brazil and identified two DNA fragments,MoT-6098 and MoT-6099,that are present in the MoT genome but not in the genome of the rice-infecting magnaporthe oryzae oryzae(MoO)*** polymerase chain reaction(PCR),we confirmed the specificity of the two markers in 53 MoT and MoO isolates from South America and *** test the efficiency of the two markers,we first established a loop-mediated isothermal amplification(LAMP)method to detect MoT at isothermal conditions,without the use of a PCR *** this,we used the Cas12a protein and guide RNAs(gRNAs)to target the MoT-6098 and MoT-6099 *** activated Cas12a showed indiscriminate single-stranded deoxyribonuclease(ssDNase)*** then combined targetdependent Cas12a ssDNase activation with recombinase polymerase amplification(RPA)and nucleic acid lateral flow immunoassay(NALFIA)to develop a method that accurately,sensitively,and cost-effectively detects MoT-specific DNA sequences in infected wheat *** novel technique can be easily adapted for the rapid detection of wheat blast and other important plant diseases in the field.

关键词： Wheat blast magnaporthe oryzae triticum Cas12a Nucleic acid rapid lateral flow immunoassay Field detection

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Ecofriendly Management of Wheat Panicle Blast Caused by magnaporthe oryzae triticum

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Agricultural Sciences 2023年第12期14卷 1751-1765页

作者： A. A. Meshuk F. M. Aminuzzaman M. R. Islam K. Nahar A. Sharmin Department of Plant Pathology Faculty of Agriculture Sher-e-Bangla Agricultural University Sher-e-Bangla Nagar Dhaka Bangladesh Department of Horticulture Faculty of Agriculture Sher-e-Bangla Agricultural University Sher-e-Bangla Nagar Dhaka Bangladesh

In this study, three wheat varieties were tested to determine seed germination and the incidence of magnaporthe oryzae triticum (MoT). Among these varieties, BARI Gom 24 (Prodip) wheat seed exhibited the highest seed germination rate (93%) but also had the highest incidence (30%) of MoT. To manage blast disease in an ecofriendly manner, seven treatments were employed: T1 = Control, T2 = Garlic clove extracts, T3 = Aloe vera leaf extracts, T4 = Black cumin seed extracts, T5 = Neem leaf extracts, T6 = Nativo 75 WG, and T7 = Provax 200 WP. The experiment was conducted using a Randomized Complete Block Design (RCBD) layout with three replications using Prodip wheat variety that exhibited highest MoT infection severity based on laboratory analysis among collected varieties. Data were collected on blast disease incidence (%), disease severity, and various growth and yield parameters of wheat. The experiment’s results indicated that among all the treatments, T7 (Seed treatment with Provax 200 WP) and T5 (Foliar spraying with Neem leaf extract) performed better in controlling blast disease in wheat. The lowest blast disease incidence (%) was observed with T7 (Provax 200 WP), with values of 7.86, 9.86, and 10.19 recorded during the milking stage, soft dough stage, and hard dough stage of wheat, respectively. T5 (Neem leaf extract) also demonstrated a statistically equivalent reduction in blast disease incidence (%). In terms of disease severity, T7 (Seed treatment with Provax 200 WP) showed the lowest values of 1.03, 1.23, and 1.63 during the milking stage, soft dough stage, and hard dough stage of wheat, respectively. Foliar spraying with neem leaf extract also exhibited similar result as of Provax 200 WP regarding panicle blast severity. As a result of these findings, it can be concluded that T5 (Neem leaf extract) is recommended as

关键词： Wheat Panicle Blast magnaporthe oryzae triticum Botanicals Management

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