Populations of Pratylenchus penetrans Relative to Decomposing Nitrogenous Soil Amendments

Populations of Pratylenchus penetrans decreased in soil following addition of 70 and 700 ppm N in the form of nitrate, nitrite, organic nitrogen, or ammonium compounds. Nitrate was less effective than other nitrogen carriers. Population reduction is principally attributed to ammonification during decomposition. This hypothesis is supported by chromatographic analyses of soil atmospheres, survival of nematodes in pure CO₂ and N₂, inverse relationship of CO₂, content in amended soils to nematode populations, and direct relationship of NH₃-N content of amended soils to nematode populations.     

Infection of Red Clover Seedlings by Heterodera trifolii Goffart and Pratylenchus penetrans (Cobb)

Penetration of 3-day-old ''Kenland'' red clover seedlings by Heterodera trifolii Goffart and Pratylenchus penetrans (Cobb) was investigated in 50-mm petri dishes on 1% agat or discs of Miracloth® (Chicopee Mills, Inc., New York, N.Y. 10018). Penetration by both nematodes increased arithmetically with increased numbers in the inoculum. H. trifolii larvae slowly penetrated all root tissue but in relatively low numbers, 25-30% of inocula. Swelling of larvae and formation of syncytia within roots was detectable 96 hr after inoculation. Initially, adults of P. penetrans preferentially penetrated the region 3-10 mm behind the root tip, but subsequently they invaded all along roots. P. penetrans penetration efficiency was high, 75-90% of inocula, and penetration was relatively rapid. When introduced simultaneously neither nematode affected the invasiveness of the other. Prior infection by H. trifolii did not affect the invasiveness of P. penetrans.     

Brassicaceous Seed Meals as Soil Amendments to Suppress the Plant-parasitic Nematodes Pratylenchus penetrans and Meloidogyne incognita

Brassicaceous seed meals are the residual materials remaining after the extraction of oil from seeds; these seed meals contain glucosinolates that potentially degrade to nematotoxic compounds upon incorporation into soil. This study compared the nematode-suppressive ability of four seed meals obtained from Brassica juncea 'Pacific Gold', B. napus 'Dwarf Essex' and 'Sunrise', and Sinapis alba 'IdaGold', against mixed stages of Pratylenchus penetrans and Meloidogyne incognita second-stage juveniles (J2). The brassicaceous seed meals were applied to soil in laboratory assays at rates ranging from 0.5 to 10.0% dry w/w with a nonamended control included. Nematode mortality was assessed after 3 days of exposure and calculated as percentage reduction compared to a nonamended control. Across seed meals, M. incognita J2 were more sensitive to the brassicaceous seed meals compared to mixed stages of P. penetrans. Brassica juncea was the most nematode-suppressive seed meal with rates as low as 0.06% resulting in > 90% suppression of both plant-parasitic nematodes. In general B. napus 'Sunrise' was the least nematode-suppressive seed meal. Intermediate were the seed meals of S. alba and B. napus 'Dwarf Essex'; 90% suppression was achieved at 1.0% and 5.0% S. alba and 0.25% and 2.5% B. napus 'Dwarf Essex', for M. incognita and P. penetrans, respectively. For B. juncea, seed meal glucosinolate-degradation products appeared to be responsible for nematode suppression; deactivated seed meal (wetted and heated at 70 °C for 48 hr) did not result in similar P. penetrans suppression compared to active seed meal. Sinapis alba seed meal particle size also played a role in nematode suppression with ground meal resulting in 93% suppression of P. penetrans compared with 37 to 46% suppression by pelletized S. alba seed meal. This study demonstrates that all seed meals are not equally suppressive to nematodes and that care should be taken when selecting a source of brassicaceous seed meal for plant-parasitic nematode management.     

Comparative Response of Alfalfa to Pratylenchus penetrans Populations

Four populations of Pratylenchus penetrans did not differ (P > 0.05) in their virulence or reproductive capability on Lahontan alfalfa. There was a negative relationship (r = -0 .7 9 ) between plant survival and nematode inocula densities at 26 ± 3 C in the greenhouse. All plants survived at an inoculum level (Pi) of 1 nematode/cm³ soil, whereas survival rates were 50 to 55% at 20 nematodes/cm³ soil. Alfalfa shoot and root weights were negatively correlated (r = - 0.87; P < 0.05) with nematode inoculum densities. Plant shoot weight reductions ranged from 13 % at Pi 1 nematode/cm³ soil to 69% for Pi 20 nematodes/cm³ soil, whereas root weight reductions ranged from 17% for Pi 1 nematode/cm³ soil to 75% for Pi 20 nematodes/cm³ soil. Maximum and minimum nematode reproduction (Pf/Pi) for the P. penetrans populations were 26.7 and 6.2 for Pi 1 and 20 nematodes/cm³ soil, respectively. There were negative correlations between nematode inoculum densities and plant survival (r = 0.84), and soil temperature and plant survival (r = -0 .7 8 ). Nematode reproduction was positively correlated to root weight (r = 0.89).     

Soil Temperature Effects on Onion Seedling Injury by Pratylenchus penetrans

The effect of soil temperatures 7-25 C upon injury to seedling onions by Pratylenchus penetrans was investigated. At 7-13 C < 100 nematodes/g of root caused significant reduction in onion root weights by the fifth week. More than 400 nematodes/g were required to produce comparable injury at 16-25 C.     

Seasonal Populations of Pratylenchus penetrans and Meloidogyne hapla in Strawberry Roots

Strawberry roots were sampled through the year to determine the populations and distribution of Pratylenchus penetrans and Meloidogyne hapla. Three strawberry root types were sampled—structural roots; feeder roots without secondary tissues; and suberized, black perennial roots. Both lesion and root-knot nematodes primarily infected feeder roots from structural roots or healthy perennial roots. Few nematodes were recovered from soil, diseased roots, or suberized roots. Lesion nematode recovery was correlated with healthy roots. In both 1997 and 1998, P. penetrans populations peaked about day 150 (end of May) and then declined. The decline in numbers corresponded to changes in total strawberry root weight and root type distribution. The loss of nematode habitat resulted from loss of roots due to disease and the transition from structural to suberized perennial roots. Meloidogyne hapla juvenile recovery peaked around 170 days (mid June) in 1997 and at 85, 147, 229, and 308 days (late March, late May, mid August, and early November, respectively) in 1998. There appear to be at least four generations per year of M. hapla in Connecticut. Diagnostic samples from an established strawberry bed may be most reliable and useful when they include feeder roots taken in late May.     

Histochemical Root Pathology of Brassica oleracea capitata L. Infected by Pratylenchus penetrans (Cobb) Filipjev and Schuurmans Stekhoyen (Nematoda: Tylenchidae)

Histochemical study of cabbage roots axenically parasitized by Pratylenchus penetrans revealed a five-fold increase in peroxidase activity (localized near lesions), pectic xylem plugs (resembling those caused by Fusarium), and accumulation of oxidase-mediated polyphenols in the region of mechanical injury. Fusarium-resistant cabbage was more susceptible to Pratylenchus than the Fusariunt-susceptible varieties, particularly in the formation of oxidized phenolic compounds. Of 13 fluorescent compounds detected by paper chromatography, one major spot was found to be ferulic acid and a minor one, catechin.     

Suppression of Pratylenchus penetrans Populations in Potato and Tomato using African Marigolds

Current strategies for management of Pratylenchus penetrans in both white potato and tomato consist of the use of fumigant or non-fumigant nematicides or crop rotation. The objective of this study was to determine if double-cropping African marigolds (Tagetes erecta) with potatoes or tomatoes could reduce P. penetrans populations. Plots were 10 m × 3 m arranged in a randomized complete block design with four replications. Treatments included marigolds, potatoes or tomatoes, and natural weedy fallow followed by either potatoes or tomatoes. Nematode populations were sampled before spring planting, between crops in August and after harvest in November. During the 3 years of the study, P. penetrans soil population density declined by an average of 93% from the pre-plant level when marigold was grown in rotation with potato and by 98% when marigold was grown.in rotation with tomato. Weedy fallow preceding potato resulted in an average decline in P. penetrans soil population density of 38%, and a similar decrease (37%) was seen when fallow preceded tomato. There was a significant reduction in the number of P. penetrans found in both potato and tomato roots when the crops followed marigolds. These results suggest that P. penetrans population density may be significantly reduced when marigolds are double-cropped with potatoes or tomatoes.     

Reproduction of Pratylenchus penetrans (Nematoda:Tylenchida)

Pratylenchus penetrans did not reproduce without males. Cytological examination indicated that cross-fertilization occurred. Females had a chromosome number of 2n = 12. Virgin females reared in isolation laid eggs, but these failed to undergo cleavage. Males reared in isolation produced sperms.     

Infection of Pratylenchus penetrans by Nematode-pathogenic fungi

Eleven fungal isolates were tested in agar dishes for pathogenicity to Pratylenchus penetrans. Of the fungi that produce adhesive conidia, Hirsutella rhossiliensis was a virulent pathogen; Verticillium balanoides, Drechmeria coniospora, and Nematoctonus sp. were weak or nonpathogens. The trapping fungi, Arthrobotrys dactyloides, A. oligospora, Monacrosporium dlipsosporum, and M. cionopagum, killed most of the P. penetrans adults and juveniles added to the fungus cultures. An isolate of Nematoctonus that forms adhesive knobs trapped only a small proportion of the nematodes. In 17-cm³ vials, soil moisture influenced survival of P. penetrans in the presence of H. rhossiliensis; nematode survival decreased with diminishing soil moisture. Hirsutella rhossiliensis and M. ellipsosporum were equally effective in reducing numbers of P. penetrans by 24-25% after 4 days in sand. After 25 days in soil artificially infested with H. rhossiliensis, numbers of P. penetrans were reduced by 28-53%.     

Interactions of Pratylenchus penetrans and Meloidogyne incognita as Coinhabitants in Tomato

Greenhouse experiments on the interactions of Pratylenchus penetrans and Meloidogyne incognita showed that the population densities of both nematode species were depressed whenthey coinhabited tomato roots. Fifty days after inoculation, the population level of a P. penetrans monoculture was about four times higher than when M. incognita was present. Conversely, M. incognita reproduced twice as fast alone as in combination. There were no significant differences in the numbers of P. penetrans when they were inoculated either 10 days prior to or after introduction of M. incognita. Root entry by P. penetrans was significantly inhibited by the presence of M. incognita. Split-root experiments showed that the inhibitory effects of M. incognita upon reproduction of P. penetrans involved factors other than the availability of feeding sites. On the other hand, the inhibitory effects of P. penetrans on M. incognita appeared to be primarily due to the quantity of available roots.     

Penetration of Celery and Alfalfa Roots by Pratylenchus penetrans as Affected by Temperature

A greater percentage of females than juveniles or males of P. penetrans penetrated celery roots grown in infested soil at 5, 18, or 30 C; the difference was greatest at 5 C. The time of initial penetration of alfalfa seedlings inoculated with single nematodes on water agar varied with temperature. Females penetrated the seedlings earlier and over a wider range of temperatures than did males or juveniles. The rate of penetration was highest for females. After initial penetration, the penetration rate decreased with time. At 13-28 C, approximately 80% of roots were penetrated by females and only 25-30% by males and juveniles by the end of the experiment.     

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.     

Histopathology of Pea Roots Axenically Infected by Pratylenchus penetrans

The histological changes in pea roots axenically infected by Pratylenchus penetrans were studied and described. Roots of pea seedlings growing aseptically on the surface of nutrient agar slants were inoculated with axenized nematodes. Six hours after inoculation most of the nematodes introduced were probing the root epidermis, but none had completely entered though a few were observed with their anterior section already in the root. Most of the nematodes penetrated the roots after 12 hr inoculation. From 18 to 24 hr after inoculation the nematodes were mostly in the mid-cortex. Invaded regions of the cortex often showed orange discoloration. As incubation continued, the number of nematodes in these roots increased, and feeding and reproductive activities extended deeper into the cortex. These activities resulted in extensive breakdown of the cortex. No nematodes were observed within the stele of infected roots; however, the endodermis of infected roots stained dark-brown. Gravid female nematodes probed the root endodermis and some endodermal cells appeared to collapse after prolonged probing by the nematode. All stages in the life cycle of the nematode were observed in infected roots; the female to male ratio inside the root was about 5:1.     

Suppression of the root-lesion nematode (Pratylenchus penetrans) in alfalfa (Medicago sativa) by Streptomyces spp.

Strains of Streptomyces were tested for their ability to reduce population densities of the root-lesion nematode (RLN), Pratylenchus penetrans, in roots of alfalfa (Medicago sativa) in growth chamber assays. Previously, these strains were shown to suppress potato scab disease, caused by Streptomyces scabies, in field experiments and to inhibit in vitrogrowth of a wide range of plant-pathogenic fungi and bacteria. Inoculation with Streptomyces at planting significantly reduced RLN population densities in roots of both susceptible and resistant alfalfa varieties grown in either heat-treated or untreated soil. Reductions in RLN population densities were observed 6 weeks after nematode inoculation. Shoot dry matter was not affected by any treatment; root dry weight was reduced in Streptomycesplus nematode treatments compared to the nematode inoculation alone in some experiments but was not affected by Streptomyces when RLN was absent. Mutant strains not producing antibiotics in vitro also reduced RLN population densities in alfalfa roots and all strains maintained high population densities after inoculation into heat-treated soil and on alfalfa roots. These strains may be useful in multi-crop, multi-pathogen management programs to augment genetic resistance to plant diseases.     

Characterization of Pratylenchus penetrans from Ten Geographically Isolated Populations Based on Their Reaction on Potato

Single female cuhures of Pratylenchus penetrans were established from soil and root samples collected from 10 geographically isolated locations in North America. The resultant isolates were used to evaluate nematode egression from and multiplication on roots of potato clones to distinguish intraspecific differences among isolates. The 10 nematode isolates were statistically separated into four groups based on percentage of nematodes that egressed from the P. penetrans-resistant potato done L 118-2. The Cornell (CR), Wisconsin (WI), Long Island (LI), and Adirondack (AD) isolates, selected as representative isolates of each of the four groups, exhibited 53%, 39%, 25%, and 10% egression from L118-2, respectively. Reproduction of these four isolates was measured on three potato cultivars (Russet Burbank, Butte, and Hudson) and two breeding lines (NY85 and L118-2). The LI and AD isolates reproduced well on all five potato clones. The CR isolate reproduced well on Russet Burbank and NY85 but significantly less on Butte, Hudson, and L118-2. Reproduction of the WI isolate was less than the LI and AD isolates but more than the CR isolate on all potato clones tested except Russet Burbank. Reproduction of the WI isolate on Russet Burbank was less than the other three isolates. Based on these results, four distinct intraspecific variants of P. penetrans are proposed: Cornell, Wisconsin, Long Island, and Adirondack.     

Effect of Hirsutella rhossiliensis on Infection of Potato by Pratylenchus penetrans

We evaluated the ability of the nematode-pathogenic fungus Hirsutella rhossiliensis (Deuteromycotina: Hyphomycetes) to reduce root penetration and population increase of Pratylenchus penetrans on potato. Experiments were conducted at 24 C in a growth chamber. When nematodes were placed on the soil surface 8 cm from a 14-day-old potato cutting, the fungus decreased the number entering roots by 25%. To determine the effect of the fungus on population increase after the nematodes entered roots, we transplanted potato cuttings infected with P. penetrans into Hirsutella-infested and uninfested soil. After 60 days, the total number of nematodes (roots and soil) was 20 ± 4% lower in Hirsutella-infested than in uninfested soil.