Genome Resource: Ralstonia solanacearum Phylotype II Sequevar 1 (Race 3 Biovar 2) Strain UW848 From the 2020 U.S. Geranium Introduction

Ralstonia solanacearum phylotype II sequevar 1 (RsII-1, formerly race 3 biovar 2) causes tomato bacterial wilt, potato brown rot, and Southern wilt of geranium. Strains in RsII-1 cause wilting in potato and tomato at cooler temperatures than tropical lowland R. solanacearum strains. Although periodically introduced, RsII-1 has not established in the United States. This pathogen is of quarantine concern and listed as a Federal Select Agent. We report a rapidly sequenced (<2 days) draft genome of UW848, a RsII-1 isolate introduced to the United States in geranium cuttings in spring 2020. UW848 belongs to the near-clonal cluster of RsII-1 global pandemic strains.

Microbial biodiversity as related to crop succession and potato intercropping for management of brown rot disease

Potato brown rot, caused by Ralstonia solanacearum, ranked globally as the second most important bacterial plant pathogen. In the present study, the influence of different cropping programs in potato brown rot management was investigated in four infected fields in Egypt. Two districts were selected as sandy soils in Giza (Wardan) and Behera (Ganuob El-Tahrir) governorates. The other two were selected as silty clay in Minufyia (Talia) and Beni-Suef (Sids) governorates. The followed crop succession included corn, potato intercropped with cabbage, onion, cowpea, wheat, corn again, and ended by potato. The pathogen was undetectable after corn, onion, and wheat. It decreased in cowpea and cabbage rhizospheres in the clay soils. The pathogen was undetectable at all districts, except at Sids, where the pathogen was significantly decreased but was not eradicated. This was possibly attributed to the high ratio of NO3− and Na+ at this district. Decreased R. solanacearum density after corn coincided with the high ratio of fluorescent pseudomonads, endospores, and actinomycetes, being most clear in the poor soils (Wardan) and less clear under iron excess at Ganoub El-Tahrir as well as the clay soils. Corn rhizosphere supported an array of antagonistic actinomycetes such as strains similar to Streptomyces intermedius, Streptomyces albidoflavus group, Streptomyces argenteolus group, and Streptomyces erythrogriseus. Intercropping potato with cabbage decreased the density of the pathogen in rhizosphere, which is associated with greater antagonistic fluorescent pseudomonads, Bacillus spp. and Serratia spp. Onion soil and rhizosphere associated with abundance of antagonists and fluorescent pseudomonads, followed by S. maltophilia and Bacillus spp. Wheat soil and rhizosphere supported fluorescent pseudomonads and antagonistic Streptomyces spp., especially in sandy soils. The pathogen was undetectable after planting the ending potato in the three districts, Wardan, Ganoub El-Tahrir, and Talia. This was accompanied by a general oligotrophism and increased ratio of fluorescent pseudomonads, endospores bacteria, and actinomycetes along with a diversity of R. solanacearum antagonists such as S. maltophilia, Citrobacter freundii, Acinetobacter sp., Delftia sp., and Serratia marcescens.

Enlargement of Gold Nanoparticles for Sensitive Immunochromatographic Diagnostics of Potato Brown Rot

Lateral flow immunoassay (LFIA) is a convenient tool for rapid field-based control of various bacterial targets. However, for many applications, the detection limits obtained by LFIA are not sufficient. In this paper, we propose enlarging gold nanoparticles’ (GNPs) size to develop a sensitive lateral flow immunoassay to detect Ralstonia solanacearum. This bacterium is a quarantine organism that causes potato brown rot. We fabricated lateral flow test strips using gold nanoparticles (17.4 ± 1.0 nm) as a label and their conjugates with antibodies specific to R. solanacearum. We proposed a signal enhancement in the test strips’ test zone due to the tetrachloroauric (III) anion reduction on the GNP surface, and the increase in size of the gold nanoparticles on the test strips was approximately up to 100 nm, as confirmed by scanning electron microscopy. Overall, the gold enhancement approach decreased the detection limit of R. solanacearum by 33 times, to as low as 3 × 104 cells∙mL–1 in the potato tuber extract. The achieved detection limit allows the diagnosis of latent infection in potato tubers. The developed approach based on gold enhancement does not complicate analyses and requires only 3 min. The developed assay together with the sample preparation and gold enlargement requires 15 min. Thus, the developed approach is promising for the development of lateral flow test strips and their subsequent introduction into diagnostic practice.

Complete Genome Sequences of the Plant Pathogens Ralstonia solanacearum Type Strain K60 and R. solanacearum Race 3 Biovar 2 Strain UW551

ABSTRACT Ralstonia solanacearum is a globally distributed plant pathogen that causes bacterial wilt diseases of many crop hosts, threatening both sustenance farming and industrial agriculture. Here, we present closed genome sequences for the R. solanacearum type strain, K60, and the cool-tolerant potato brown rot strain R. solanacearum UW551, a highly regulated U.S. select agent pathogen.