Evaluation of Natural and Factitious Food Sources for Pronematus ubiquitus on Tomato Plants

Pronematus ubiquitus (McGregor) is a small iolinid mite that is capable of establishing on tomato plants. Once established, this mite has been shown to control both tomato russet mite, Aculops lycopersici (Tryon) (Acari: Eriophyidae), and tomato powdery mildew (Oidium neolycopersici L. Kiss). In the present study, we explored the effects of a number of alternative food sources on the oviposition rate in the laboratory. First, we assessed the reproduction on food sources that P. ubiquitus can encounter on a tomato crop: tomato pollen and powdery mildew, along with tomato leaf and Typha angustifolia L. In a second laboratory experiment, we evaluated the oviposition rate on two prey mites: the astigmatid Carpoglyphus lactis L. (Acari: Carpoglyphidae) and the tarsonemid Tarsonemus fusarii Cooreman (Acari: Tarsonemidae). Powdery mildew and C. lactis did not support reproduction, whereas tomato pollen and T. fusarii did promote egg laying. However, T. angustifolia pollen resulted in a higher oviposition in both experiments. In a greenhouse trial on individual caged tomato plants, we evaluated the impact of pollen supplementation frequency on the establishment of P. ubiquitus. Here, a pollen addition frequency of every other week was required to allow populations of P. ubiquitus to establish.

Preliminary test: Evaluation and selection of tomato (Lycopersicon esculentum Mill.) varieties resistant to drought and powdery mildew

Powdery mildew (Leveillula taurica) and (Oidium neolycopersici) are two harmful fungi that invade the tomato (Lycopersicon esculentum Mill.) plant and grow in dry conditions. Under the influence of polyethylene glycol 6000 at a concentration of 12%, the total seedlings fresh weight, total seedlings dry weight, seedling growth rate of one seedling, seedling length, and tissue water content percentage for three tomato varieties were assessed. Despite the superiority of the (Mobil) tomato variety in terms of numerical values, the results revealed no significant differences between varieties. As a result, (Mobil) has greater vigor under environmental drought stress of lower osmotic stress than other tomato varieties. More work is required to evaluate the research selection of varieties resistant to biotic stresses in dry areas, such as powdery mildew disease.

Evaluation of Natural and Factitious Food Sources for Pronematus Ubiquitus on Tomato Plants

Pronematus ubiquitus (McGregor) is a small iolinid mite that is capable of establishing on tomato plants. Once established, this mite has been shown to control both tomato russet mite, Aculops lycopersici (Tryon) (Acari: Eriophyidae) and tomato powdery mildew (Oidium neolycopersici L. Kiss). In the present study, we explored the nutritional value of various food sources in the laboratory. First, we assessed the reproduction of two food sources that P. ubiquitus can encounter on a tomato crop: tomato pollen and powdery mildew. In a second laboratory experiment, we evaluated the nutritional value of two types of prey mites: the astigmatid Carpoglyphus lactis L. (Acari: Carpoglyphidae) and the tarsonemid Tarsonemus fusarii (Acari: Tarsonemidae). Powdery mildew and C. lactis did not contribute to the reproduction, whereas tomato pollen and T. fusarii did allow egg-laying. However, Typha angustifolia pollen was a superior food source in both experiments. In a greenhouse trial on individual caged tomato plants, we evaluated the impact of pollen supplementation frequency on establishment of P. ubiquitus. Here, a pollen addition frequency of every other week was required to allow populations of P. ubiquitus to establish.

Discovery and Characterization of a Novel Tomato mlo Mutant from an EMS Mutagenized Micro-Tom Population

In tomato (Solanum lycopersicum), there are at least three SlMLO (Mildew resistance Locus O) genes acting as susceptibility genes for the powdery mildew disease caused by Oidium neolycopersici, namely SlMLO1, SlMLO5 and SlMLO8. Of the three homologs, the SlMLO1 gene plays a major role since a natural mutant allele called ol-2 can almost completely prevent fungal penetration by formation of papillae. The ol-2 allele contains a 19-bp deletion in the coding sequence of the SlMLO1 gene, resulting in a premature stop codon within the second cytoplasmic loop of the predicted protein. In this study, we have developed a new genetic resource (M200) in the tomato cv. Micro-Tom genetic background by means of ethyl methane sulfonate (EMS) mutagenesis. The mutant M200 containing a novel allele (the m200 allele) of the tomato SlMLO1 gene showed profound resistance against powdery mildew with no fungal sporulation. Compared to the coding sequence of the SlMLO1 gene, the m200 allele carries a point mutation at T65A. The SNP results in a premature stop codon L22* located in the first transmembrane domain of the complete SlMLO1 protein. The length of the predicted protein is 21 amino acids, while the SlMLO1 full-length protein is 513 amino acids. A high-resolution melting (HRM) marker was developed to distinguish the mutated m200 allele from the SlMLO1 allele in backcross populations. The mutant allele conferred recessive resistance that was associated with papillae formation at fungal penetration sites of plant epidermal cells. A comprehensive list of known mlo mutations found in natural and artificial mutants is presented, which serves as a particularly valuable resource for powdery mildew resistance breeding.

ShNPSN11, a vesicle-transport-related gene, confers disease resistance in tomato to Oidium neolycopersici

Tomato powdery mildew, caused by Oidium neolycopersici, is a fungal disease that results in severe yield loss in infected plants. Herein, we describe the function of a class of proteins, soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs), which play a role in vesicle transport during defense signaling. To date, there have been no reports describing the function of tomato SNAREs during resistance signaling to powdery mildew. Using a combination of classical plant pathology-, genetics-, and cell biology-based approaches, we evaluate the role of ShNPSN11 in resistance to the powdery mildew pathogen O. neolycopersici. Quantitative RT-PCR analysis of tomato SNAREs revealed that ShNPSN11 mRNA accumulation in disease-resistant varieties was significantly increased following pathogen, compared with susceptible varieties, suggesting a role during induced defense signaling. Using in planta subcellular localization, we demonstrate that ShNPSN11 was primarily localized at the plasma membrane, consistent with the localization of SNARE proteins and their role in defense signaling and trafficking. Silencing of ShNPSN11 resulted in increased susceptibility to O. neolycopersici, with pathogen-induced levels of H2O2 and cell death elicitation in ShNPSN11-silenced lines showing a marked reduction. Transient expression of ShNPSN11 did not result in the induction of a hypersensitive cell death response or suppress cell death induced by BAX. Taken together, these data demonstrate that ShNPSNl11 plays an important role in defense activation and host resistance to O. neolycopersici in tomato LA1777.

Development of DNA markers for Slmlo1.1, a new mutant allele of the powdery mildew resistance gene SlMlo1 in tomato (Solanum lycopersicum)

Reductions in growth and quality due to powdery mildew (PM) disease cause significant economic losses in tomato production. Oidium neolycopersici was identified as the fungal species responsible for tomato PM disease in South Korea in the present study, based on morphological and internal transcribed spacer DNA sequence analyses of PM samples collected from two remote regions (Muju and Miryang). The genes involved in resistance to this pathogen in the tomato accession ‘KNU-12’ (Solanum lycopersicum var. cerasiforme) were evaluated, and the inheritance of PM resistance in ‘KNU-12’ was found to be conferred via simple Mendelian inheritance of a mutant allele of the PM susceptibility locus Ol-2 (SlMlo1). Full-length cDNA analysis of this newly identified mutant allele (Slmlo1.1) showed that a 1-bp deletion in its coding region led to a frameshift mutation possibly resulting in SlMlo1 loss-of-function. An alternatively spliced transcript of Slmlo1.1 was observed in the cDNA sequences of ‘KNU-12’, but its direct influence on PM resistance is unclear. A derived cleaved amplified polymorphic sequence (dCAPS) and a high-resolution melting (HRM) marker were developed based on the 1-bp deletion in Slmlo1.1, and could be used for efficient marker-assisted selection (MAS) using ‘KNU-12’ as the source for durable and broad-spectrum resistance to PM.