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Validation of HLK effector-based DNA marker for diagnosis of Ralstonia solanacearum infecting solanaceous vegetables.

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Title Validation of HLK effector-based DNA marker for diagnosis of Ralstonia solanacearum infecting solanaceous vegetables.
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Creator B. R. Meena, A N Tripathi and Sujan Majumder
 
Subject effector
 
Description Not Available
NRalstonia solanacearum (Solanaceous wilt pathogen) is the second most important bacterial plant pathogen. Detection and identification of associated pathogens are of prime importance for disease management which emphasizes the need for efficient diagnostic tests and specific detection tools. Molecular techniques are most specific, highly sensitive time saving, less laborious and can simultaneously detect and identify the plant pathogens. A multi-gene family encoding HLK effectors one of the virulence determinants is well conserved in all Ralstonia solanacearum strains indicating their role in fitness of the pathogen was identified for development of specific DNA marker for diagnosis. The primer set (hlk1Fand hlk1R) was designed based on sequence alignment of hlk1 effector genes from 15 different bacterial strain sequences downloaded from NCBI data base and primer oligonucleotide were synthesised. The PCR conditions were optimised to amplify the specific fragment of HLK1 effector gene sequence. The primers were able to amplify size of 203 nucleotides with annealing temperature of 57 oC for Ralstonia solanacearum which was further validated for detection across the isolates of wilt pathogen from different host including eggplant, chilli, potato, cluster bean, marigold and ginger. To check the specificity of primers to Ralstonia solanacearum the amplification attempts were made with four bacterial species (Psuedomonas putida biovar 4, Psuedomonas aurigenosa , Xnathomonas oryzae, Bacillus cibi) and two fungal species (Fusarium oxysporium and Alternaria solani). We were unable to amplify DNA from above mentioned microorganisms which further validates the specificity of primers to Ralstonia solanacearum. The diagnostic tool developed will be helpful to assess bacterial survival, dissemination and disease outbreaks for effective management of bacterial wilt in solanaceous crops.
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Date 2020-09-11T10:04:50Z
2020-09-11T10:04:50Z
2019-10-11
 
Type Presentation
 
Identifier B. R. Meena, A N Tripathi and Sujan Majumder, ICAR-Indian Institute of Vegetable Research 2019. “Validation of HLK effector-based DNA marker for diagnosis of Ralstonia solanacearum infecting solanaceous vegetables. “presentation in the International seminar on “Sustainable Agricultural Development in Changing Global Scenario” to be held on 11-13th October 2019 at IESD, BHU Varanasi, UP
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http://krishi.icar.gov.in/jspui/handle/123456789/41198
 
Language English
 
Relation Not Available;
 
Publisher Royal Association for Science-led Socio-cultural Advancement (RASSA)