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Transgenic Resistance Confers Effective Field Level Control of Bacterial Spot Disease in Tomato

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Abstract

We investigated whether lines of transgenic tomato (Solanum lycopersicum) expressing the Bs2 resistance gene from pepper, a close relative of tomato, demonstrate improved resistance to bacterial spot disease caused by Xanthomonas species in replicated multi-year field trials under commercial type growing conditions. We report that the presence of the Bs2 gene in the highly susceptible VF 36 background reduced disease to extremely low levels, and VF 36-Bs2 plants displayed the lowest disease severity amongst all tomato varieties tested, including commercial and breeding lines with host resistance. Yields of marketable fruit from transgenic lines were typically 2.5 times that of the non-transformed parent line, but varied between 1.5 and 11.5 fold depending on weather conditions and disease pressure. Trials were conducted without application of any copper-based bactericides, presently in wide use despite negative impacts on the environment. This is the first demonstration of effective field resistance in a transgenic genotype based on a plant R gene and provides an opportunity for control of a devastating pathogen while eliminating ineffective copper pesticides.

Citation: Horvath DM, Stall RE, Jones JB, Pauly MH, Vallad GE, et al. (2012) Transgenic Resistance Confers Effective Field Level Control of Bacterial Spot Disease in Tomato. PLoS ONE 7(8): e42036. doi:10.1371/journal.pone.0042036

Received: May 25, 2012; Accepted: June 29, 2012; Published: August 1, 2012

Copyright: © Horvath et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
IntroductionBacterial spot disease, a complex of four Xanthomonas species, is among the most widespread and destructive diseases of tomatoes and peppers throughout the world, causing lesions on aerial plant parts leading to defoliation and fruit loss [1]. It has chronically afflicted U.S. tomato production, particularly in Florida, where the largest production of fresh market tomatoes occurs. Ninety-seven per cent of Florida acres are affected, and yield losses may reach fifty per cent of marketable production [2].

Various crop protection compounds have been used to control bacterial spot in commercial tomato and pepper production. In the 1950s, streptomycin was commonly used to control plant diseases caused by bacteria, including bacterial spot. Xanthomonas euvesicatoria (Race T1), the prevalent bacterial spot race in Florida at the time, quickly became resistant to streptomycin [3], [4], and its use was discontinued. In the 1960s, fixed copper compounds and copper-fungicide mixes became the primary means of bacterial spot control. Initially fixed copper was used alone, but resistance in xanthomonads developed quickly [5]. In response to observing increased efficacy when copper was mixed with ethylenebisdithiocarbamate (EBDC) fungicides such as maneb and mancozeb, growers began mixing fixed copper products with EBDC fungicides for improved bacterial spot control [5]. However, even these copper-fungicide mixes have become ineffective against tomato races of the pathogen, especially under conditions of high humidity and warm temperatures that favor heavy disease pressure (eg [6], [7], [8], [9]; Table S1).

Because crop protection compounds do not control the copper tolerant Xanthomonas races responsible for this disease, genetic resistance against bacterial spot has been a priority in tomato breeding programs. These breeding efforts have been slowed by the complex genetics of resistance and changing races of the pathogen, and consequently there are no commercial varieties with effective resistance to Xanthomonas. One commonly bred form of plant disease resistance relies on the evolution of specific intracellular immune receptors encoded by disease resistance, orR, genes. R genes have been selected through conventional breeding for over 100 years [10]. They encode specific receptors that recognize gene products made by specific races of a given pathogen species. These pathogen components are termed effectors, and they contribute to pathogen virulence by suppressing or modulating host defenses in susceptible plant genotypes that lack a corresponding R gene [11].

AvrBs2 is an effector that is highly conserved in a number of Xanthomonas species that infect a wide range of plant hosts, including tomato [12]. Unlike other key effectors, it is present in all six races of the tomato bacterial spot disease complex (Table S2). Mutations in AvrBs2 can impair virulence, indicating that it plays an important role in pathogenicity and may be a good target for durable resistance [12].

AvrBs2 is recognized by the R protein Bs2, identified in pepper, a fellow member of the Solanaceae and close relative of tomato. In the current study, we tested whether Bs2 transgenic tomato lines were effective against current field races of bacterial spot in multi-year field trials conducted in two commercial growing regions of Florida. Our results demonstrated that tomatoes carrying the Bs2gene and which received no bactericidal crop protection compounds had the highest disease resistance of all the genotypes tested and had significantly increased yields relative to control.

Continues @ PLOS ONE: Transgenic Resistance Confers Effective Field Level Control of Bacterial Spot Disease in Tomato:


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