Michael E. Matheron

Activity of Boscalid, Fenhexamid, Fluazinam, Fludioxonil, and Vinclozolin on Growth of Sclerotinia minor and S. sclerotiorum and Development of Lettuce Drop

Sclerotinia drop is a major disease of lettuce caused by two soilborne fungi, Sclerotinia minor and S. sclerotiorum. Fungicides such as dicloran (Botran), iprodione (Rovral), and vinclozolin (Ronilan) are currently available in the United States to manage this disease. Studies were conducted to investigate the relative effect of some new fungicides, including boscalid, fenhexamid, fluazinam, and fludioxonil, in comparison with vinclozolin, on growth of S. minor and S. sclerotiorum in agar plate tests as well as control of lettuce drop in the field. At a rate of 0.001 μg/ml, all tested compounds only suppressed mycelial growth of either pathogen from 0 to 20%. At 0.01 μg/ml, mycelial growth of S. minor was reduced 82 to 84% by fludioxonil and fluazinam and only 1 to 16% by boscalid, fenhexamid, and vinclozolin. At the same rate, mycelial growth of S. sclerotiorum was reduced 78% by fluazinam and from 0 to 12% by boscalid, fludioxonil, fenhexamid, and vinclozolin. At 0.1 μg/ml, all tested chemistries except vinclozolin inhibited mycelial growth of S. minor from 70 to 98%, whereas growth of S. sclerotiorum was suppressed 95 to 99% by fludioxonil and fluazinam, significantly less (40 to 47%) by boscalid and fenhexamid, and not at all by vinclozolin. At a rate of 1.0 μg/ml, all tested fungicides reduced mycelial growth of S. minor and S. sclerotiorum from 87 to 100% and 77 to 100%, respectively. Mycelial growth emerging from sclerotia of S. minor was reduced from 98 to 100% by all fungicides tested at a rate of 1.0 μg/ml, whereas growth from sclerotia of S. sclerotiorum was suppressed from 90 to 96% by fenhexamid, fludioxonil, fluazinam, and vinclozolin. In lettuce plots infested with S. minor, boscalid and fluazinam provided the highest level of disease control, significantly greater than that achieved with fenhexamid, fludioxonil, and vinclozolin. In the presence of S. sclerotiorum, the highest degree of disease suppression occurred with application of fluazinam, fludioxonil, and vinclozolin, whereas the least effective compound was fenhexamid. Boscalid and fluazinam were more effective against lettuce drop caused by S. minor than disease caused by S. sclerotiorum.

Impact of Azoxystrobin, Dimethomorph, Fluazinam, Fosetyl-Al, and Metalaxyl on Growth, Sporulation, and Zoospore Cyst Germination of Three Phytophthora spp.

In vitro activity of azoxystrobin, dimethomorph, and fluazinam on growth, sporulation, and zoospore cyst germination of Phytophthora capsici, P. citrophthora, and P. parasitica was compared to that of fosetyl-Al and metalaxyl. The 50% effective concentration (EC50) values for)inhibition of mycelial growth of the three pathogens usually were lowest for dimethomorph and (metalaxyl, ranging from <0.1 to 0.38 μg/ml. However, the 90% effective concentration (EC90) levels for dimethomorph always were lower than the other four tested compounds, with values ranging from 0.32 to 1.6 μg/ml. Mycelial growth of P. capsici, P. citrophthora, and P. parasitica was least affected by azoxystrobin and fluazinam, with estimated (EC90) values >3,000 μg/ml. Reduction of sporangium formation by P. capsici, P. citrophthora, and P. parasitica in the presence of dimethomorph at 1 μg/ml was significantly greater than that recorded for the same concentration of azoxystrobin, fluazinam, and fosetyl-Al. For the three species of Phytophthora, zoospore motility was most sensitive to fluazinam (EC50 and EC90 values of <0.001 μg/ml) and (least sensitive to fosetyl-Al, with (EC50 and EC90 values ranging from 299 to 334 and 518 to 680 μg/ml, respectively). Germination of encysted zoospores of P. capsici, P. citrophthora, and P. parasitica was most sensitive to dimethomorph (EC50 and EC90 values ranging from 3.3 to 7.2 and 5.6 to 21 μg/ml, respectively), intermediate in sensitivity to fluazinam (EC50 and EC90 from 18 to 108 and 67 to >1,000 μg/ml, respectively) and metalaxyl (EC50 and EC90 from 32 to 280 and 49 to 529 μg/ml, respectively), and lowest in sensitivity to azoxystrobin and fosetyl-Al (EC50 and EC90 from 256 to >1,000 μg/ml). The activity of azoxystrobin, dimethomorph, and fluazinam on one or more stages of the life cycle of P. capsici, P. citrophthora, and P. parasitica suggests that these compounds potentially could provide Phytophthora spp. disease control comparable to that of the established fungicides fosetyl-Al and metalaxyl.

Suppression of Phytophthora root and crown rot on pepper plants treated with acibenzolar-S-methyl

The fungicide mefenoxam is registered for the control of Phytophthora blight of peppers caused by Phytophthora capsici. Isolates of the pathogen that are insensitive to mefenoxam, however, have been detected in some locations. Consequently, alternative methods are needed to control Phytophthora blight of peppers. Acibenzolar-S-methyl (ABM, Actigard) is a chemical activator of plant disease resistance that has potential for the management of Phytophthora blight of peppers. The effect of foliar applications of ABM on the development of root and crown rot on pepper plants grown in the greenhouse and inoculated with Phytophthora capsici or in soil naturally infested with the pathogen was evaluated. Inhibition of stem canker development on pepper cvs. Bell Tower and AZ9 after four treatments with ABM (75 μg/ml) was significantly greater than on plants receiving a single application of the chemical. Stem canker length on Bell Tower or AZ9 peppers was inhibited by 93.2 to 97.2% and 87.4 to 92.4% when plants were inoculated with P. capsici at 1 or 5 weeks, respectively, after the fourth application of ABM. Survival of chile pepper plants grown in field soil naturally infested with P. capsici was significantly increased by three foliar applications of ABM (75 μg/ml) compared with nontreated plants in all three trials when pots were watered daily and in two of three trials when pots were flooded for 48 h every 2 weeks. When soil was flooded every 2 weeks to establish conditions highly favorable for disease development, plants treated once with mefenoxam (100 μg/ml) survived significantly longer than those treated with ABM. On the other hand, when water was provided daily without periodic flooding to establish conditions less favorable for disease development, plant survival between the two chemicals was not different in two of three trials. Length of survival among chile pepper plants treated twice with 25, 50, or 75 μg/ml of ABM and grown in soil infested with P. capsici was not different. This work indicates that ABM could be an important management tool for Phytophthora root and crown rot on pepper plants.

Comparison of Five Fungicides on Development of Root, Crown, and Fruit Rot of Chile Pepper and Recovery of Phytophthora capsici from Soil

The activity of five fungicides, azoxystrobin, dimethomorph, fluazinam, fosetyl-Al, and metalaxyl (subsequently replaced with mefenoxam by the manufacturer), was compared for effects on the development of root, crown, and fruit rot of chile pepper and on recovery of Phytophthora capsici from naturally infested soil. When inoculated with zoospores, plants survived longer and shoot and root fresh weights were greater for plants drenched with metalaxyl at 10 μg/ml than for plants treated with the same rate of azoxystrobin or dimethomorph. At 100 μg/ml, the duration of plant survival was greater for dimethomorph and fluazinam than for azoxystrobin; however, shoot and root growth did not differ. In soil naturally infested with P. capsici, survival and growth of shoots and roots for plants treated with dimethomorph at 100 μg/ml were greater than for those treated with the same rate of azoxystrobin or fluazinam. The most effective compounds for inhibition of lesion development on stems and fruit were mefenoxam at 1,200 μg/ml and dimethomorph at 480 μg/ml. Recovery of P. capsici from soil treated with each of the five tested compounds was significantly less than that recorded for soil not receiving a fungicide. The potential and relative value of azoxystrobin, dimethomorph, fosetyl-Al, and fluazinam as chemical management tools for Phytophthora blight on chile pepper, in addition to metalaxyl (replaced with mefenoxam), has been demonstrated.

First Report of Fusarium Wilt of Lettuce Caused by Fusarium oxysporum f. sp. lactucae in Arizona

A new wilt and root rot disease was observed in 6 and 11 commercial fields of lettuce (Lactuca sativa) in western Arizona during the fall of 2001 and 2002, respectively. Distance between infested sites ranged from approximately 0.5 to 39 km. Five head lettuce cultivars as well as a red leaf lettuce cultivar were affected. Disease symptoms included yellowing and wilting of leaves, as well as stunting and plant death. The cortex of the crown and upper root of infected plants usually was decayed and reddish brown. Disease symptoms first appeared at the time of plant thinning and continued to develop up to plant maturity. Fusarium oxysporum was consistently isolated from symptomatic plant roots. Seeds of cv. Lighthouse were planted in nonsterile vermiculite within 3.0-cm-square × 7.0-cm-deep cells in a transplant tray and thinned to a single plant per cell. When the first true leaves were emerging, 10 individual seedlings were inoculated with a single-spore isolate of F. oxysporum recovered from diseased lettuce root cortex tissue. Inoculum was prepared by growing the fungus on potato dextrose agar in 100-mm-diameter × 15-mm-deep plastic petri dishes at 28°C with a 12-h photoperiod under fluorescent light. Once the fungus completely covered the agar surface, 50 ml of sterile distilled water was added to the dish, and the mycelia and conidia on the surface were scraped off the agar and suspended in the water. This fungal suspension was decanted, and a 2-ml aliquot containing 1.8 × 105 CFU was pipetted into the vermiculite near the stem of each lettuce seedling. Ten plants grown in noninfested vermiculite served as uninoculated controls. After inoculation, plants were maintained in a growth chamber at 28°C with a 12-h photoperiod under fluorescent light for 3 weeks. Symptoms of yellowing, wilt, vascular decay, and often plant death developed during the incubation period on all inoculated plants but not on control plants. Fusarium oxysporum was consistently reisolated from inoculated plants but not from uninoculated plants. The experiment was repeated and yielded the same results. A wilt and root rot disease of lettuce attributed to F. oxysporum f. sp. lactucae was first reported in Japan in 1967 (3) and subsequently in the United States (San Joaquin Valley of California) in 1993 (2), and Italy in 2002 (1). The researchers of the U.S. report did not cite the earlier work from Japan and described the pathogen as F. oxysporum f. sp. lactucum. The Arizona isolate used to demonstrate pathogenicity was of the same vegetative compatibility group as an isolate of the pathogen from lettuce in California reported in 1993. Several companies grow and harvest lettuce in Arizona and California. At the end of production and harvest in the fall, tractors, implements, and harvesting equipment are transported from the San Joaquin Valley in California to western Arizona. The similarity between the isolate of F. oxysporum f. sp. lactucae from western Arizona and the San Joaquin Valley of California suggest a possible introduction of the pathogen into Arizona from California, perhaps on soil adhering to farm equipment. To our knowledge, this is the first report of F. oxysporum f. sp. lactucae infecting lettuce in Arizona. References: (1) A. Garibaldi et al. Plant Dis. 86:1052, 2002. (2) J. C. Hubbard and J. S. Gerik. Plant Dis. 77:750, 1993. (3) T. Matuo and S. Motohashi. Trans. Mycol. Soc. Jpn. 8:13, 1967.

Influence of Soil Temperature and Moisture on Eruptive Germination and Viability of Sclerotia of Sclerotinia minor and S. sclerotiorum

The effect of soil temperature and moisture on eruptive germination and viability of sclerotia of Sclerotinia minor and S. sclerotiorum in field soil was examined. In two trials at constant temperatures, the proportion of sclerotia of both pathogens that germinated in wet soil ( ≥-0.02 MPa) tended to decrease as soil temperature increased from 15 to 40°C, with no germination of sclerotia of S. minor and S. sclerotiorum detected after 1 and 2 weeks, respectively, at 40°C. In contrast, after 1 to 4 weeks in dry soil ( ≤-100 MPa) at 40°C, germination of sclerotia of S. minor and S. sclerotiorum ranged from 28 to 55% and 42 to 77%, respectively. In field trials, the germination rate of sclerotia of S. minor and S. sclerotiorum after 2 to 8 weeks in irrigated soil on the surface or buried at a depth of 5 cm was significantly lower than that for sclerotia maintained in dry soil at the same depths. On the other hand, after burial at a depth of 10 cm, germination of sclerotia in irrigated and dry soil did not differ significantly after 2 to 8 weeks for S. minor and after 2, 4, and 8 weeks for S. sclerotiorum. For both pathogens, germination of sclerotia from 2 to 8 weeks in irrigated soil with a mean temperature of 32°C was significantly lower than that for sclerotia in irrigated soil with a mean temperature of 26°C. In microplot trials conducted in July and August, no sclerotia of S. minor and S. sclerotiorum germinated after 2 and 3 weeks, respectively, after recovery from flooded soil with mean soil temperatures ranging from 30 to 33°C. A flood irrigation is often applied to fields for salt management during July or August in the Yuma lettuce production region. Results from these studies suggest that maintaining this flooding event for 2 to 3 weeks in fields with a history of lettuce drop caused by S. minor and S. sclerotiorum could significantly reduce the population of viable sclerotia.

Biocontrol of Lettuce Drop Caused by Sclerotinia sclerotiorum and S. minor in Desert Agroecosystems

Field experiments were conducted over 2 years in Yuma County, AZ, and Imperial County, CA, to determine the efficacy of several biocontrol agents for the management of lettuce drop caused by Sclerotinia spp. Commercial formulations of Trichoderma harzianum (Plantshield, Supersivit), Gliocladium virens (Soilgard), Coniothyrium minitans (Contans), and Bacillus subtilis (Companion) were evaluated and compared with the chemical fungicide iprodione (Rovral) against Sclerotinia sclerotiorum and S. minor. A single application of biocontrol products or of Rovral did not reduce lettuce drop caused by either Sclerotinia species. However, two applications of Contans, one at planting and one at post-thinning, significantly reduced the incidence of lettuce drop caused by S. sclerotiorum and increased yield but had no effect on S. minor at both locations in both years. Two applications of other biocontrol products did not significantly reduce disease incidence despite medium to high recovery following application. In contrast, Contans was only sporadically recovered following application. In vitro fungicide sensitivity evaluation revealed that both Trichoderma and Gliocladium species were tolerant to iprodione, dicloran (Botran), and vinclozolin (Ronilan) up to 1,000 ppm a.i., whereas both Sclerotinia spp. and C. minitans were sensitive to all three fungicides above 1 ppm. In summary, Contans was the most effective treatment for the control of lettuce drop caused by S. sclerotiorum, but no treatment was effective against S. minor in the desert lettuce production systems.

Effect of planting date, cultivar, and stage of plant development on incidence of fusarium wilt of lettuce in desert production fields

Fusarium wilt of lettuce, first recognized in Japan in 1955, has since been discovered in the United States (California in 1990, Arizona in 2001), Iran (1995), Taiwan (1998), and Italy (2001). In Arizona, the causal agent, Fusarium oxysporum f. sp. lactucae, has been recovered from lettuce plants in 27 different lettuce fields during the 2001 to 2003 production seasons. Studies were initiated to examine the impact of planting date, cultivar, and stage of plant development on the incidence of disease in the field. In 2002 and 2003, tested lettuce cultivars were sown in at least one of the following planting windows; early-season (September), mid-season (October), and late-season (December). Within each planting window, significant differences in disease incidence among lettuce cultivars were noted at plant maturity. The mean incidence of Fusarium wilt on cultivars sown in September, October, and December was 92.3, 15.1, and 2.0%, respectively, in 2002 and 74.2, 5.1, and 0.7%, respectively, in 2003. The mean soil temperatures at the10-cm depth during the September, October, and December plantings for both years were 26, 14, and 14°C, respectively. Initial symptoms of Fusarium wilt were apparent as early as 14 days after seeding, with increasing incidence of disease noted as the crop developed and reached maturity. Among all lettuce cultivars planted in September, only one and two cultivars of romaine in 2002 and 2003, respectively, reached maturity with ≤5% incidence of Fusarium wilt, whereas the lowest incidence of disease among crisphead, green leaf, red leaf, or butterhead cultivars was 73.7, 27.0, 20.2, and 65.7%, respectively, in 2002 and 62.1, 29.0, 100, and 100%, respectively, in 2003. For October plantings, all romaine cultivars had ≤5% incidence of Fusarium wilt at maturity, whereas disease incidence among tested cultivars of crisphead lettuce in 2002 and 2003 ranged from 0.8 to 66.8% and 0.3 to 43.3%, respectively. When planted in December, 82 and 88% of tested cultivars, including all romaine entries, reached maturity with ≤1% incidence of Fusarium wilt. Selection of appropriate lettuce cultivars and planting times should allow successful production of lettuce in the southwestern Arizona production region with minimal or no incidence of disease in fields infested with F. oxysporum f. sp. lactucae. On the other hand, successful production of lettuce in infested fields when temperatures favor disease development will not be possible until lettuce cultivars are developed that possess high tolerance or resistance to the pathogen.

Resistance to Phytophthora citrophthora and P. parasitica and Nursery Characteristics of Several Citrus Rootstocks

Studies were conducted to compare existing and potential citrus rootstocks with respect to resistance to root rot and gummosis caused by Phytophthora citrophthora and P. parasitica in greenhouse and growth chamber experiments and horticultural performance under simulated nursery conditions. Depending upon rootstock and experiment, mean root weights resulting from inoculation with P. citrophthora were 27 to 96% lower than the comparable controls. In similar experiments with the same rootstocks, inoculation with P. parasitica resulted in root weights that were 38 to 95% less than weights of the noninoculated controls. During 1994 or 1995, mean root weight reduction compared with noninoculated plants among Citrus macrophylla, rough lemon, C. volkameriana, and Sunki mandarin × Flying Dragon trifoliate (62-109-19) attributable to P. citrophthora and mean root weight reduction among C. macrophylla, C. volkameriana, rough lemon, Sacaton citrumelo, Sunki mandarin × Flying Dragon trifoliate (62-109-19), African shaddock × Rubidoux trifoliate, and Shekwasha mandarin × English trifoliate attributable to P. parasitica were significantly less than those recorded for all other tested rootstocks. Rootstocks that sustained a low percentage of root weight reduction generally experienced a low percentage of shoot weight reduction and survived longer as well. In evaluation of resistance to gummosis, depending on rootstock and experiment, the mean length of stem lesions caused by P. citrophthora on rootstocks ranged from 0.2 to 25.0 mm, whereas values for P. parasitica ranged from 0.2 to 18.5 mm. Stem lesions smaller than 5 mm in length were recorded for 21 and 14 of 36 different rootstocks inoculated with P. citrophthora and P. parasitica, respectively. On the other hand, P. citrophthora and P. parasitica caused stem lesions of at least 10 mm in length on 8 and 16 citrus rootstocks, respectively. Desirable nursery characteristics, including vigorous growth, minimal branching, and high leaf chlorophyll content, were demonstrated most prominently by Gomiri rough lemon, C. volkameriana, and Benton citrange, and to a lesser degree by some other rootstocks. Possible factors that could account for inconsistent classification of some citrus rootstocks as susceptible or resistant to Phytophthora root rot and gummosis are discussed.

Evaluation of Soil Solarization and Flooding As Management Tools for Fusarium Wilt of Lettuce

Fusarium wilt of lettuce caused by Fusarium oxysporum f. sp. lactucae continues to spread and cause economic losses in Arizona lettuce fields since the initial discovery of the disease in the state in 2001. Studies were initiated to assess the potential of summer soil solarization and flooding as management tools for Fusarium wilt of lettuce in southwestern Arizona production fields. In microplot studies, lettuce plant growth in soil naturally infested with F. oxysporum f. sp. lactucae that was solarized from 2 to 8 weeks was consistently greater than growth in nonsolarized soil. Growth of lettuce in flooded soil containing the pathogen occasionally was significantly higher than in nonflooded soil; however, the effect on plant growth and health was not as consistent as that recorded for solarized soil. In four trials within a field containing F. oxysporum f. sp. lactucae, the incidence of Fusarium wilt on lettuce sown in soil after solarization was reduced from 42 to 91% compared with disease in nonsolarized plots. There was no significant benefit of a 2- over a 1-month solarization period under the conditions of these trials, where the mean soil temperature at a depth of 5 cm during a 1-month solarization period in 2005 and 2006 was 47 and 49°C, respectively. These findings suggest that soil solarization can be an effective tool for management of Fusarium wilt on lettuce, especially when used within an integrated program in conjunction with existing disease management tactics.