Molecular Detection of Ralstonia solanacearum to Facilitate Breeding for Resistance to Bacterial Wilt in Potato.

Abstract

Potato bacterial wilt is caused by the devastating bacterial pathogen Ralstonia solanacearum. Quantitative resistance to this disease has been and is currently introgressed from a number of wild relatives into cultivated varieties through laborious breeding programs. Here, we present two methods that we have developed to facilitate the screening for resistance to bacterial wilt in potato. The first one uses R. solanacearum reporter strains constitutively expressing the luxCDABE operon or the green fluorescent protein (gfp) to follow pathogen colonization in potato germplasm. Luminescent strains are used for nondestructive live imaging, while fluorescent ones enable precise pathogen visualization inside the plant tissues through confocal microscopy. The second method is a BIO-multiplex-PCR assay that is useful for sensitive and specific detection of viable R. solanacearum (IIB-1) cells in latently infected potato plants. This BIO-multiplex-PCR assay can specifically detect IIB-1 sequevar strains as well as strains belonging to all four R. solanacearum phylotypes and is sensitive enough to detect without DNA extraction ten bacterial cells per mL in complex samples.The described methods allow the detection of latent infections in roots and stems of asymptomatic plants and were shown to be efficient tools to assist potato breeding programs.

Plant phenotyping relevance

ジャガイモの細菌病抵抗性スクリーニング用に、病原体レポーター株による非破壊ライブイメージング・共焦点可視化法を開発しており、植物体内の感染・病徴状態の取得が中心的な方法論的貢献です。PCR法単独なら分子診断ですが、本研究は画像ベースの感染表現型評価法も明確に含みます。

Code and data availability

This is a methods/protocol book chapter describing potato bacterial wilt resistance screening and Ralstonia detection procedures. No public phenotype datasets, plant images, analysis code, trained models, or data supplements are mentioned; no availability or deposit statements appear in the supplied blocks.

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