Incidence and Molecular Identification of Cochliobolus carbonum as Causal Organism of Rice Seedling Blight

During last few years, seedling blight and black lesions along culm of rice seedlings have been appearing frequently in nurseries. Identification of the causal organism was the objective of this research. The identification was done based on morpho-pathological characteristics and Internal Transcrib...

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Main Authors: Rabie A.S. El-Shafey, Kotb A. Attia, Fatma A. Mostafa, Rabab M. Elamawi
Format: Article
Language:English
Published: SpringerOpen 2018-12-01
Series:Beni-Suef University Journal of Basic and Applied Sciences
Online Access:http://www.sciencedirect.com/science/article/pii/S2314853517303700
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author Rabie A.S. El-Shafey
Kotb A. Attia
Fatma A. Mostafa
Rabab M. Elamawi
author_facet Rabie A.S. El-Shafey
Kotb A. Attia
Fatma A. Mostafa
Rabab M. Elamawi
author_sort Rabie A.S. El-Shafey
collection DOAJ
description During last few years, seedling blight and black lesions along culm of rice seedlings have been appearing frequently in nurseries. Identification of the causal organism was the objective of this research. The identification was done based on morpho-pathological characteristics and Internal Transcribed Spacer (ITS) region sequencing analysis. The pathogen effects on germination and development of rice seedlings were studied under laboratory and greenhouse conditions. Based on the morphological examination, 10 isolates were identified as Cochliobolus carbonum fungus. This organism recorded different levels of pathogenicity, and most of the isolates showed high virulence, up to 56% as seedling infection with isolate ECC-5. The germination percentage was reduced to 73% for cultivar Sakha 101. Sakha 101 recorded highest infected seedlings percentage (44%) under artificial inoculation. The isolates’ reaction was more variable on seedlings than on leaves of Egyptian rice cultivars. Infection severity on leaves of isolate ECC-5 recorded highest average for infection, while isolate ECC-3 did not exhibit any infection. Most of C. carbonum isolates were induced severe infection on rice seedlings. Therefore, the identified isolates were recorded as a novel pathogen causing seedling blight disease on rice. Soaking in crude extract filtrate of C. carbonium isolates culture reduced germination percentage to 1.55% with Sakha101 and inhibit the shoots and roots growth. The molecular variation using ISJ markers reflected a high level of genetic variation between isolates. The ITS region sequencing of two isolates ECC-7 and ECC-9 was successfully analyzed, and alignment with 19 isolates of Bipolaris zeicola worldwide with 97% identity. Phylogenetic analyses of sequences resulted in a well-resolved phylogeny. The data suggested that ITS region analysis was a potential tool for phylogenetic reconstruction of the new isolates and as was DNA barcode for identification of the fungal species. It confirmed that this organism is a rice seed-borne pathogen that causes seedling blight disease. Keywords: C. carbonum, Rice seedling blight, Internal Transcribed Spacer (ITS) region sequencing analysis
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spelling doaj.art-9153679139524cf0a3bfbc86f8bc5e7f2022-12-22T00:51:10ZengSpringerOpenBeni-Suef University Journal of Basic and Applied Sciences2314-85352018-12-0174652662Incidence and Molecular Identification of Cochliobolus carbonum as Causal Organism of Rice Seedling BlightRabie A.S. El-Shafey0Kotb A. Attia1Fatma A. Mostafa2Rabab M. Elamawi3Rice Research and Training Center, Field Crop Research Institute, Agricultural Research Center, Sakha, Kafr El-Sheikh, 33717, EgyptRice Research and Training Center, Field Crop Research Institute, Agricultural Research Center, Sakha, Kafr El-Sheikh, 33717, EgyptIntegrated Pest Management Department, Plant Pathology Research Institute, Agricultural Research Center, Giza, EgyptRice Pathology Department, Plant Pathology Research Institute, Agricultural Research Center, Sakha, Kafr El-Sheikh, 33717, Egypt; Corresponding author.During last few years, seedling blight and black lesions along culm of rice seedlings have been appearing frequently in nurseries. Identification of the causal organism was the objective of this research. The identification was done based on morpho-pathological characteristics and Internal Transcribed Spacer (ITS) region sequencing analysis. The pathogen effects on germination and development of rice seedlings were studied under laboratory and greenhouse conditions. Based on the morphological examination, 10 isolates were identified as Cochliobolus carbonum fungus. This organism recorded different levels of pathogenicity, and most of the isolates showed high virulence, up to 56% as seedling infection with isolate ECC-5. The germination percentage was reduced to 73% for cultivar Sakha 101. Sakha 101 recorded highest infected seedlings percentage (44%) under artificial inoculation. The isolates’ reaction was more variable on seedlings than on leaves of Egyptian rice cultivars. Infection severity on leaves of isolate ECC-5 recorded highest average for infection, while isolate ECC-3 did not exhibit any infection. Most of C. carbonum isolates were induced severe infection on rice seedlings. Therefore, the identified isolates were recorded as a novel pathogen causing seedling blight disease on rice. Soaking in crude extract filtrate of C. carbonium isolates culture reduced germination percentage to 1.55% with Sakha101 and inhibit the shoots and roots growth. The molecular variation using ISJ markers reflected a high level of genetic variation between isolates. The ITS region sequencing of two isolates ECC-7 and ECC-9 was successfully analyzed, and alignment with 19 isolates of Bipolaris zeicola worldwide with 97% identity. Phylogenetic analyses of sequences resulted in a well-resolved phylogeny. The data suggested that ITS region analysis was a potential tool for phylogenetic reconstruction of the new isolates and as was DNA barcode for identification of the fungal species. It confirmed that this organism is a rice seed-borne pathogen that causes seedling blight disease. Keywords: C. carbonum, Rice seedling blight, Internal Transcribed Spacer (ITS) region sequencing analysishttp://www.sciencedirect.com/science/article/pii/S2314853517303700
spellingShingle Rabie A.S. El-Shafey
Kotb A. Attia
Fatma A. Mostafa
Rabab M. Elamawi
Incidence and Molecular Identification of Cochliobolus carbonum as Causal Organism of Rice Seedling Blight
Beni-Suef University Journal of Basic and Applied Sciences
title Incidence and Molecular Identification of Cochliobolus carbonum as Causal Organism of Rice Seedling Blight
title_full Incidence and Molecular Identification of Cochliobolus carbonum as Causal Organism of Rice Seedling Blight
title_fullStr Incidence and Molecular Identification of Cochliobolus carbonum as Causal Organism of Rice Seedling Blight
title_full_unstemmed Incidence and Molecular Identification of Cochliobolus carbonum as Causal Organism of Rice Seedling Blight
title_short Incidence and Molecular Identification of Cochliobolus carbonum as Causal Organism of Rice Seedling Blight
title_sort incidence and molecular identification of cochliobolus carbonum as causal organism of rice seedling blight
url http://www.sciencedirect.com/science/article/pii/S2314853517303700
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