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Molecular assays to detect and quantify DNA from viable cells of the seedborne pathogen Xanthomonas hortorum pv. carotae in carrot seed were developed and evaluated for use on nontreated and hot-water-treated seed lots. Both a TaqMan real-time polymerase chain reaction (PCR) assay and a loop-mediated isothermal amplification (LAMP) dilution endpoint assay detected and quantified DNA from viable pathogen cells after treatment of carrot seed washes with the livedead discriminating dye propidium monoazide (PMA). The detection limits of the assays were approximately 10¹ CFU for pure cultures of X. hor-torum pv. carotae, and 10² to 10³ CFU/g seed from naturally infested carrot seed lots. X. hortorum pv. carotae in and on carrot seed was killed by soaking the seed in hot water (52°C for 25 min), and a subsequent PMA treatment of these hot-water-treated seed washes suppressed detection of the pathogen with both the real-time PCR and LAMP assays. For 36 commercial seed lots treated with PMA but not hot water, regression of colony counts of X. hortorum pv. carotae measured by dilution plating on a semiselective agar medium versus estimates of pathogen CFU determined by the molecular assays resulted in significant (P < 0.05) linear relationships (R2 = 0.68 for the real-time PCR assay and 0.79 for the LAMP assay). The molecular assays provided quantitative estimates of X. hortorum pv. carotae infestations in carrot seed lots in <24 h, which is a significant improvement over the 7 to 14 days required to obtain results from the traditional dilutionplating assay. [ABSTRACT FROM AUTHOR]

Cladosporium variabile, Stemphylium botryosum, and Verticillium dahliae are seedborne and seed-transmitted pathogens of spinach. Spinach seed treatments in 1.2% NaOCl for 10 to 60 rain, or hot water (40, 45, 50, 55, and 60°C) for 10 to 40 min, were evaluated for eradication of these fungi from seed. C. variabile and V. dahliae were largely eradicated by chlorine treatment for ≥10 min. Although chlorine treatment reduced the incidence of S. botryosum, this fungus was not eradicated after 60 min in chlorine. Seed germination was not affected adversely by chlorine treatment, even after 60 min. In contrast, germination was reduced significantly by hot water treatment at 50°12 for ≥30 min or 55 or 60°C for ≥10 min. C. variabile was eradicated from seed treated in 40°C water for 10 min. V dahliae was eradicated from seed treated at 55°C for ≥30 rain or 60°C for ≥10 min. S. botryosum was eradicated from a lightly infected seed lot (5% incidence) by hot water treatment at 55 or 60% for ≥10 min, but could not be eradicated from two heavily infected lots (»65% incidence), even at 60°C for 40 min. Using precisely controlled parameters, chlorine or hot water seed treatments can be used to eradicate C. variabile and reduce the incidence of S. botryosum and V. dahliae in spinach seed without damaging germination. [ABSTRACT FROM AUTHOR]

Two pathogenicity groups of Fusarium oxysporum f. sp. spinaciae, the causal agent of Fusarium wilt of spinach (Spinacia oleracea), were described recently based on virulence of isolates on proprietary spinach inbreds. In this study, a wide range in severity of wilt was observed for 68 spinach cultivars inoculated with an isolate of each pathogenicity group, with 22 (32.4%) cultivars displaying differential responses to the isolates. In a second set of trials, seven spinach cultivars were inoculated with five isolates of each pathogenicity group. The cultivars had similar wilt responses to isolates within each group. In both sets of trials, the most severe wilt developed on cultivars inoculated with pathogenicity group 2 isolates when daylength was shorter and light intensity lower. To test whether light intensity exacerbates severity of Fusarium wilt, three spinach cultivars were inoculated with two isolates of each pathogenicity group and grown with or without shading. Shaded plants developed more severe wilt than nonshaded plants. This difference in wilt severity was greatest for plants inoculated with pathogenicity group 2 isolates. We propose naming isolates of pathogenicity groups 1 and 2 as races 1 and 2 of F. oxysporum f. sp. spinaciae, respectively, and recommend the cultivars Kiowa (susceptible to both races) and Magnetic (susceptible to race 2 and highly resistant to race 1) as differentials. Results of this study should help breeders screen spinach germplasm for resistance to both races of F. oxysporum f. sp. spinaciae. [ABSTRACT FROM AUTHOR]

The efficacy of 14 seed and drench treatments for control of soilborne damping-off pathogens in organic production of spinach was evaluated in a greenhouse study. The efficacy of each treatment was compared with nontreated seed and seed treated with a conventional fungicide for control of Fusariurn oxysporuin f. sp. spinaciae, Pythiurn ultimurn, and Rhizoctonia solani. Two experimental seed treatments, GTG I and GTG II (each comprised of a proprietary organic disinfectant and the latter also containing Trichoderma harzianum T22), provided equivalent control to the conventional fungicide, mefenoxam, against P. ultimum in one trial and significant reduction of damping-off in the second trial. Natural II and Natural X (Streptomycele products), and Subtilex (Bacillus subtilis) seed treatments each suppressed damping-off significantly in one of the two trials. For R. solani, GTG I and Natural It seed treatments reduced damping-off as effectively as a drench with the fungicide Terraclor (pentachloronitrobenzene). A soil drench with Prestop (Gliocladium catenulatuin) suppressed postemergence wilt caused by F oxysporum in both trials; a compost tea drench and seed treatment with Yield Shield (Bacillus pumilis) each suppressed postemergence wilt in only one of two trials. GTG I and GTG II significantly increased seed germination compared to nontreated seed. No treatment was effective against all three pathogens, and some treatments exacerbated damping-off. [ABSTRACT FROM AUTHOR]