Stem Nematode-Fusarium Wilt Complex in Alfalfa as Related to Irrigation Management at Harvest Time

A high moisture level in the top 10 cm of soil at time of cutting of alfalfa increased the incidence of plant mortality and Fusarium wilt in soil infested with Ditylenchus dipsaci and Fusarium oxysporum f. sp. medicaginis in greenhouse and field microplot studies. Ranger alfalfa, susceptible to both D. dipsaci and F. oxysporum f. sp. medicaginis, was less persistent than Moapa 69 (nematode susceptible and Fusarium wilt resistant) and Lahontan alfalfa (nematode resistant with low Fusarium wilt resistance). In the greenhouse, the persistence of Ranger, Moapa 69, and Lahontan alfalfa plants was 46%, 64%, and 67% respectively, in nematode + fungus infested soil at high soil moisture at time of cutting. This compared to 74%, 84%, and 73% persistence of Ranger, Moapa 69, and Lahontan, respectively, at low soil moisture at time of cutting. Shoot weights as a percentage of uninoculated controls at the high soil moisture level were 38%, 40%, and 71% for Ranger, Moapa 69, and Lahontan, respectively. Low soil moisture at time of cutting negated the effect D. dipsaci on plant persistence and growth of subsequent cuttings, and reduced Fusarium wilt of plants in the nematode-fungus treatment; shoot weights were 75%, 90%, and 74% of uninoculated controls for Ranger, Moapa 69, and Lahontan. Similar results were obtained in the field microplot study, and stand persistence and shoot weights were less in nematode + fungus-infested soil at the high soil-moisture level (early irrigation) than at the low soil-moisture level (late irrigation).     

Effects of Environmental Factors and Cultural Practices on Parasitism of Alfalfa by Ditylenchus dipsaci

Cool humid weather enhanced development and reproduction of Ditylenchus dipsaci in alfalfa in laboratory and field studies in Utah. Relative humidity and nematode reproduction were positively correlated (P < 0.05), whereas air temperature and nematode reproduction were negatively correlated (P < 0.05). The greatest number of nematodes per gram of alfalfa tissue was found in nondormant Moapa alfalfa tissue at St. George during April, whereas the greatest numbers of nematodes were found in dormant Ranger alfalfa in June at West Jordan and Smithfield. There was 100% invasion of both resistant Lahontan and susceptible Ranger alfalfa plants at soil moisture levels of 61-94% field capacity. Fall burning of alfalfa to control weeds reduced, and spring burning increased, the incidence of invaded plants, nematodes per gram of plant tissue, and the mortality of susceptible Ranger (P < 0.01) and Moapa (P < 0.01) alfalfa plants over that of plants in nonburned control plots. Fall burning also reduced and spring burning increased the incidence of invaded plants (P < 0.05), but had no influence on nematodes per gram of plant tissue or the mortality of resistant Lahontan and Nevada Synthetic XX alfalfa over those of plants in control plots.     

Relationship of Ditylenchus dipsaci and Harvest Practices to the Persistence of Alfalfa

Persistence of dormant Ranger and nondormant Moapa alfalfas, both susceptible to Ditylenchus dipsaci, varied with stand age and cutting frequency. Stand reduction increased with cutting frequency. In D. dipsaci-infested soil, stand reductions in Ranger 1, 4, and 5 years old exceeded reductions in stands 2 and 3 years old; persistence was greatest in 2-year-old stands. In Moapa alfalfa, D. dipsaci reduced stands the most in years 2 and 3; whereas persistence was greatest in 1-year-old stands. Harvesting Ranger alfalfa one, two, three, and four times during the growing season reduced 2-year-old stands by 10, 14, 19, and 29% in D. dipsaci-infested soil and by 2, 4, 4, and 7% in uninfested soil, respectively. Comparable reductions in Moapa alfalfa were 13, 16, 18, and 38% in infested soil and 0, 2, 4, and 6% in uninfested soil. Cutting frequency had less effect on persistence of resistant semidormant Lahontan grown in D. dipsaci-infested soil relative to susceptible cultivars. Increasing the number of cuttings per year decreased storage of total nonstructural carbohydrate and adversely affected persistence of alfalfa stands and yields; the greatest negative effects occurred on both resistant and susceptible alfalfa in D. dipsaci-infested soil.     

Suppression of Alfalfa Growth by Concommitant Populations of Pratylenchus penetrans and two Fusarium species

Growth of alfalfa (Medicago sativa cv. Vernal) seedlings was compared after inoculation with combinations of either Pratylenchus penetrans and Fusarium soloni or P. penetrans and F. oxysporum f. sp. medicaginis. A synergistic disease interaction occurred in alfalfa when F. oxysporum and P. penetrans were added simultaneously to the soil. Alfalfa growth was suppressed at all inoculum levels of P. penetrans and F. oxysporum, but not with F. solani. Seedlings inoculated with the nematode alone gave lower yields than when inoculated with either Fusarium species alone. Fusarium oxysporum, but not F. solani, was pathogenic to alfalfa under similar experimental conditions. Fusarium oxysporum did not alter the populations of P. penetrans in alfalfa roots, whereas the presence of F. solani was associated with a diminished number of P. penetrans in the roots.     

A Nematicide Seed Treatment to Control Ditylenchus dipsaci on Seedling Alfalfa

Three nematicides were evaluated as seed treatments to control the alfalfa stem nematode (Ditylenchus dipsaci) on seedling alfalfa. Alfalfa seeds were soaked for 10 hours in a 0.5% (formulated by weight) concentration of either carbofuran, phenamiphos or oxamyl in acetone with no adverse effect on seed germination. All three treatments decreased nematode damage and increased survival of ''Ranger'' (susceptible) and ''Lahontan'' (resistant) alfalfa plants, when seeds were planted in soil infested with D. dipsaci. Mean live plant counts after 6 weeks in the untreated control, acetone alone, carbofuran, phenamiphos, and oxamyl treatments, respectively, were 4.3, 6.3, 19.0, 19.8, and 19.0 for Lahontan and 4.5, 1.5, 18.5, 19.3, and 18.0 for Ranger from 20 seeds/pot. Nematicide seed treatments resulted in significantly healthier Ranger alfalfa plants 4 months after planting. The combination of seed treatment and host resistance may provide a means of establishing alfalfa in an alfalfa monocropped system where soil populations of D. dipsaci are high.     

Differential Reaction of Alfalfa Cultivars to Meloidogyne hapla and M. chitwoodi Populations

Meloidogyne hapla reproduced and suppressed growth (P < 0.05) of susceptible Lahontan and Moapa alfalfa at 15, 20, and 25 C. At 30 C, resistant Nevada Syn XX lost resistance to M. hapla. M. hapla invaded and reproduced on Rhizobium meliloti nodules of Lahontan and Moapa, inducing giant cell formation and structural disorder of vascular bundles of nodules without disrupting bacteroids. At 15, 20, and 25 C a M. chitwoodi population from Utah reproduced on Lahontan, Moapa, and Nevada Syn XX alfalfa, suppressing growth (P < 0.05). Final densities of the Utah M. chitwoodi population were greater (P < 0.05) than those of Idaho and Washington State populations on Lahontan at 15 and 25 C and on Nevada Syn XX at 15 C, but were less consistent and smaller (P < 0.05) than those of M. hapla on Lahontan and Moapa at 20 and 25 C. Inconsistent reproduction of the Utah M. chitwoodi population on alfalfa suggests the possible existence of nematode strains revealed by variability in alfalfa resistance. No reproduction or inconsistent final nematode population densities with no damage were observed on Lahontan, Moapa, and Nevada Syn XX plants grown in soil infested with Idaho and Washington State M. chitwoodi populations.     

Interaction of Fusarium oxysporum f. sp. ciceri and Meloidogyne javanica on Cicer arietinum

Interaction of Meloidogyne javanica and Fusarium oxysporum f. sp. ciceri was studied on Fusarium wilt-susceptible (JG 62 and K 850) and resistant (JG 74 and Avrodhi) chickpea cultivars. In greenhouse experiments, inoculation of M. javanica juveniles prior to F. oxysporum f. sp. ciceri caused greater wilt incidence in susceptible cultivars and induced vascular discoloration in roots of resistant cultivars. Nematode reproduction was greatest (P = 0.05) at 25 °C. Number of galls and percentage of root area galled increased when the temperature was increased from 15 °C to 25 °C. Wilt incidence was greater at 20 °C than at 25 °C. Chlorosis of leaves and vascular discoloration of plants did not occur at 15 °C. The nematode enhanced the wilt incidence in wilt-susceptible cultivars only at 25 °C. Interaction between the two pathogens on shoot and root weights was significant only at 20 °C, and F. o. ciceri suppressed the nematode density at this temperature. Wilt incidence was greater in clayey (48% clay) than in loamy sand (85% sand) soils. The nematode caused greater plant damage on loamy sand than on clayey soil. Fusarium wilt resistance in Avrodhi and JG 74 was stable in the presence of M. javanica across temperatures and soil types.     

Early Root Response to Meloidogyne incognita in Resistant and Susceptible Alfalfa Cultivars

The early events of Meloidogyne incognita behavior and associated host responses following root penetration were studied in resistant (cv. Moapa 69) and susceptible (cv. Lahontan) alfalfa. Ten-day-old seedlings of alfalfa cultivars were inoculated with second-stage juveniles (J2) and harvested 12, 24, 48, and 72 hours and 7, 14, and 21 days later. Both cultivars supported similar root penetration and initial J2 migration. By 72 hours after inoculation the majority of J2 were amassed inside the vascular cylinder in roots of susceptible Lahontan, while J2 had not entered the vascular cylinder of resistant Moapa 69 and remained clumped at the root apex. Nematode development progressed normally in Lahontan, but J2 were not observed in Moapa 69 after day 7. The greatest differences between RNA translation products isolated from inoculated and uninoculated roots of Lahanton occurred 72 hours after inoculation. Only minor differences in gene expression were observed between inoculated and uninoculated Moapa 69 roots at 72 hours. Comparison of translation products from inoculated versus mechanically wounded Lahontan roots revealed products that were specific to or enhanced in nematode-infected plants. Moapa 69 appears to possess a type of resistance to M. incognita that does not depend on a conventional hypersensitive response.     

Biology and Pathogenicity of Pratylenchus neglectus on Alfalfa

Pratylenchus neglectus reduced the growth of alfalfa cultivars in greenhouse and growth chamber studies. Inocula (1,000, 5,000 and 10,000 nematodes per plant) reduced shoot dry weights of Ranger by 16, 27, and 40%, of Lahontan by 16, 32, and 40%, and of Nevada Synthetic XX (Nev Syn XX) by 18, 26, and 37%, respectively, at 26 ñ 2 C. Pratylenchus neglectus at 1,000 nematodes per plant reduced Ranger shoot dry weights by 5, 12, 18, and 27%, at 15, 20, 25, and 30 C, respectively, whereas 5,000 nematodes per plant reduced shoot dry weights by 12, 17, 26, and 38%, respectively, at similar temperatures. Reductions in dry root weights were directly related to reductions in shoot growth. At 1,000 nematodes per plant, Ranger root dry weights were reduced by 3, 14, 40, and 40%, whereas 5,000 nematodes per plant reduced root dry weight by 25, 31, 59, and 63%, respectively, at similar temperatures. Similar results were observed on Lahontan and Nev Syn XX at the same inoculum levels and soil temperatures. Nematode reproductive indices (final nematode population per plant divided by initial nematode inoculum per plant) were higher at 1,000 nematodes per plant than at 5,000 nematodes per plant, were positively correlated with temperature, and were unaffected by cultivar.     

Interaction between Meloidogyne incognita and Fusarium oxysporum f. sp. phaseoli on Selected Bean Genotypes

Four bean genotypes (IPA-1, A-107, A-211, and Calima), representing all possible combinations of resistance and susceptibility to Fusarium oxysporum f. sp. phaseoli (Fop) and Meloidogyne incognita, were each inoculated with three population densities of these pathogens. Calima and A-107 were resistant to Fop; A-107 and A-211 were resistant to M. incognita; and IPA-1 was susceptible to both pathogens. In Fop-susceptible lines (IPA-1 and A-211), the presence of M. incognita contributed to an earlier onset and increased severity of Fusarium wilt symptoms and plant stunting. However, the Fop-resistant Calima developed symptoms of Fusarium wilt only in the presence of M. incognita. Genotype A-107 (resistant to both M. incognita and Fop) exhibited Fusarium wilt symptoms and a moderately susceptible reaction to Fop only after the breakdown of its M. incognita resistance by elevated incubation temperatures (27 C). Root galling and reproduction of M. incognita was generally increased as inoculum density of M. incognita was increased on the M. incognita susceptible cultivars. However, these factors were decreased as the inoculum density of Fop was increased. It was concluded that severe infections of bean roots by M. incognita increase the severity of Fusarium wilt on Fop-susceptible genotypes and may modify the resistant reaction to Fop.     

The Influence of Trichoderma harzianum on the Root-knot Fusarium Wilt Complex in Cotton

Wilt-susceptible cultivar ''Rowden'' cotton was inoculated wilh Meloidogyne incognita (N), Trichoderma harzianum (T), and Fusarium oxysporum f. sp. vasinfectum (F) alone and in all combinations in various time sequences. Plants inoculated with F alone or in combination with T did not develop wilt, Simultaneous inoculation of 7-day-old seedlings with all three organisms (NTF) produced earliest wilt. However, plants receiving nematodes at 7 days and Fusarium and Trichoderma at 2 or 4 weeks later (N-T-F, N-TF) developed the greatest wilt between 49-84 days after initial nematode inoculation. During the same period, Fusarium added 4 weeks after initial nematode inoculation (N-F) and Fusarium added 4 weeks after initial simultaneous inoculation of nematode and Trichoderma (NT-F) produced the least wilt. The addition of Fusarium inhibited nematode reproduction. Simultaneous inoculation with nematodes and Trichoderma (NT-) resulted in the greatest root gall development, whereas nematodes alone produced the greatest number of larvae. In comparison with noninoculated controls (CK), treatments involving all three organisms inhibited plant growth, plants inoculated with the nematode alone (N-) or with nematodes and Trichoderma (NT-) simultaneously had greatest root weight. Any treatment involving the nematode resulted in fewer bolls per plant and greater necrosis on roots than the noninoculated checks.     

Influence of Soil Fumigation on the Fusarium-Root-knot Nematode Disease Complex of Cotton in California

For control of the root-knot nematode, Meloidogyne incognita, and the pathogenic wilt fungus, Fusarium oxysporum, on cotton, soil fumigants were applied in the field at conventional and higher rates. Conventional rates suppressed Fusarium wilt but higher rates gave quicker early growth, better stands, less stand loss over the season, a lower percentage of plants infected with wilt, fewer plants with vascular discoloration, and fewer nematodes. The best treatment about doubled the yields of untreated controls in one experiment and quadrupled them in another.     

Effect of combination of bio-agents and mineral nutrients for the management of alfalfa wilt pathogen Fusarium oxysporum f. sp. medicaginis

Abstract

Biological and nutrient management of soil borne disease is increasingly gaining stature as a possible practical and safe approach. Inhibitory effects of fungal and bacterial antagonists were tested under in vitro conditions against the wilt pathogen of alfalfa Fusarium oxysporum f. sp. medicaginis. Trichoderma harzianum and Pseudomonas fluorescens (PI 5) were found to be effective against the alfalfa wilt pathogen. Manganese sulphate at 500 and 750 ppm inhibited the mycelial growth of F. oxysporumf. sp. medicaginis under in vitro conditions. In pot culture studies, manganese sulphate at 12.5 mg/kg reduced the wilt incidence (23.33%). Combined application of manganese sulphate 12.5 mg/kg + T. harzianum 1.25 mg/kg of soil significantly reduced the wilt incidence accompanied by improved plant growth and yield in pot culture. The mixture of manganese sulphate (25 kg/ha) + T. harzianum (2.5 kg/ha) significantly reduced the wilt incidence when applied as a basal dose in the field conditions. The average mean of disease reduction was 62.42% over control.     

Evaluation of Hot Water Treatments for Management of Ditylenchus dipsaci and Fungi in Daffodil Bulbs

Treatment of daffodil (Narcissus pseudonarcissus) bulbs in a 0.37% formaldehyde water solution at 44 C for 240 minutes is a standard practice in California for management of the stem and bulb nematode, Ditylenchus dipsaci. Recent concern over the safety of formaldehyde and growers'' requests for a shorter treatment time prompted a reevaluation of the procedure. The time (Y, in minutes) required to raise the temperature at the bulb center from 25 to 44 C was related to bulb circumference (X, in cm) and is described by the linear regression Y = -15 + 3.4X. The time required for 100% mortality of D. dipsaci in vitro without formaldehyde was 150, 60, and 15 minutes at 44, 46, and 48 C, respectively. Hot water treatment (HWT) with 0.37% formaldehyde at 44 C for 150 minutes controlled D. dipsaci and did not have a detrimental effect on plant growth and flower production. Shorter formaldehyde-HWT of 90, 45, and 30 minutes at 46, 48, and 50 C, respectively, controlled D. dipsaci but suppressed plant growth and flower production. Fungal genera commonly isolated from the bulbs in association with D. dipsaci were Penicillium sp., Fusarium oxysporum f. sp. narcissi, and Mucor plumbeus, representing 60, 25, and 5%, respectively, of the total fungi isolated. These fungi caused severe necrosis in daffodil bulbs. HWT at 44 C for 240 minutes reduced the number of colonies recovered from bulbs. The effects of formaldehyde, glutaraldehyde, and sodium hypochlorite in reducing the population of fungi within bulbs were variable. Satisfactory control of D. dipsaci within bulbs can be achieved with HWT of bulbs at 44 C for 150 minutes with 0.37% formaldehyde or at 44 C for 240 minutes without chemicals.     

Interaction of Population Levels of Fusarium oxysporum f. sp. vasinfectum and Meloidogyne incognita on Cotton

In autoclaved greenhouse soil without Fusarium oxysporum f. sp. vasinfectum, Meloidogyne incognita did not cause leaf or vascular discoloration of 59-day-old cotton plants. Plants had root galls with as few as 50 Meloidogyne larvae per plant. Root galling was directly proportional to the initial nematode population level. Fusarium wilt symptoms occurred without nematodes with 77,000 fungus propagules or more per gram of soil. As few as 50 Meloidogyne larvae accompanying 650 fungus propagules caused Fusarium wilt. With few exceptions, leaf symptoms appeared sooner as numbers of either or both organisms increased. In soils infested with both organisms, the extent of fungal invasion and colonization was well correlated with the extent of nematode galling and other indications of the Fusarium wilt syndrome.     

Interactions of Concomitant Species of Nematodes and Fusarium oxysporum f. sp. vasinfectum on Cotton

Meloidogyne incognita, Hoplolaintus galeatus, and North Carolina and Georgia populations of Belonolaimus longicaudatus were introduced singly and in various combinations with Fusarium oxysporum f. sp. vasinfectum on wilt-susceptible ''Rowden'' cotton. Of all the nematodes, the combination of the N. C. population of B. longicaudatus with Fusarium promoted greatest wilt development. H. galeatus had no effect on wilt. With Fusarium plus M. incognito or B. longicaudatus, high nematode levels promoted greater wilt than low levels. The combination of either population of B. longicaudatus with M. incognita and Fusarium induced greater wilt development than comparable inoculum densities of either nematode alone or where H. galeatus was substituted for either of these nematodes. Nematode reproduction was inversely related to wilt development. Without Fusarium, however, the high inoculum level resulted in greater reproduction of all nematode species on cotton. Combining M. incognita with B. longicaudatus or H. galeatus gave mutually depressive effects on final nematode populations. The interactions of H. gateatus with B. longicaudatus varied with two populations of the latter.     

Association of Criconemella xenoplax and Fusarium spp. with Root Necrosis and Growth of Peach

Criconemella xenoplax, Fusarium solani, and F. oxysporum caused necrosis of Nemaguard peach feeder roots in greenhouse tests. Root necrosis was more extensive in the presence of either fungus than wtih C. xenoplax alone. Shoot growth and plant height were less for plants inoculated with F. oxysporum or F. solani than for plants inoculated with the fungi plus C. xenoplax. Neither synergistic nor additive effects on root necrosis or plant growth occurred between C. xenoplax and the fungal pathogens.     

Predisposition of Broadleaf Tobacco to Fusarium Wilt by Early Infection with Globodera tabacum tabacum or Meloidogyne hapla

In greenhouse experiments, broadleaf tobacco plants were inoculated with tobacco cyst (Globodera tabacum tabacum) or root-knot (Meloidogyne hapla) nematodes 3, 2, or 1 week before or at the same time as Fusarium oxysporum. Plants infected with nematodes prior to fungal inoculation had greater Fusarium wilt incidence and severity than those simultaneously inoculated. G. t. tabacum increased wilt incidence and severity more than did M. hapla. Mechanical root wounding within 1 week of F. oxysporum inoculation increased wilt severity. In field experiments, early-season G. t. tabacum control by preplant soil application of oxamyl indirectly limited the incidence and severity of wilt. Wilt incidence was 48%, 23%, and 8% in 1989 and 64%, 60%, and 19% in 1990 for 0.0, 2.2, and 6.7 kg oxamyl/ha, respectively. Early infection of tobacco by G. t. tabacum predisposed broadleaf tobacco to wilt by F. oxysporum.     

Plantago lanceolata and Plantago rugelii Extracts are Toxic to Meloidogyne incognita but not to Certain Microbes

Extracts from the plants Plantago lanceolata and P. rugelii were evaluated for toxicity to the root-knot nematode Meloidogyne incognita, the beneficial microbes Enterobacter cloacae, Pseudomonas fluorescens and Trichoderma virens, and the plant-pathogenic fungi Fusarium oxysporum f. sp. gladioli, Phytophthora capsici, Pythium ultimum, and Rhizoctonia solani. Wild plants were collected, roots were excised from shoots, and the plant parts were dried and ground to a powder. One set of extracts (10% w/v) was prepared in water and another in methanol. Treatments included extract concentrations of 25%, 50%, 75% and 100%, and water controls. Meloidogyne incognita egg hatch was recorded after 7-day exposure to the treatments, and second-stage juvenile (J2) activity after 48 hours. All extracts were toxic to eggs and J2, with P. lanceolata shoot extract tending to have the most activity against M. incognita. Numbers of active J2 remained the same or decreased in a 24-hour water rinse following the 48-hour extract treatment, indicating that the extracts were lethal. When data from water- and methanol-extracted roots and shoots of both plant species were combined for analysis, J2 tended to be more sensitive than eggs to the toxic compounds at lower concentrations, while the higher concentrations (75% and 100%) were equally toxic to both life stages. The effective concentrations causing 50% reduction (EC₅₀) in egg hatch and in J2 viability were 44.4% and 43.7%, respectively. No extract was toxic to any of the bacteria or fungi in our assays.