Anhydrobiosis in Pratylenchus penetrans.

Anhydrobiotic survival of Pratylenchus penetrans was compared in several soil moisture regimes. Bodies of anhydrobiotic nematodes were coiled. In slow-dried soils, Vineland silt loam (VSL) and Fox loamy sand (FLS), 70 and 58% of the total P. penetrans populations were anhydrobiotic when soil moistures reached ca. 3% and water potential 15 kPa or greater. Coiling began at a much lower water potential in FLS than in VSL. In fast-dried soils, only 31 and 22% of the P. penetrans populations in the same two soil types had entered the anhydrobiotic state at comparable moistures. In the above soils, 76-96% of the P. penetrans were alive immediately after entering the anhydrobiotic state. In slow-dried VSL, some nematodes (1%) survived 770 days. In the other soils, all anhydrobiotic nematodes were dead after 438 days. Anhydrobiosis increased the ability of nematodes to survive subzero temperatures, but it did not increase their ability to survive temperatures above 40 C. Infectivity and reproductivity of rehydrated P. penetrans were not affected by anhydrobiosis.

Morphological Observations of Pratylenchus penetrans from Celery and Strawberry in Southern Ontario.

Pratylenchus penetrans was obtained from Premier strawberry in Norfolk County and the Niagara Peninsula and from celery in the latter area. Host affected the dimensions of P. penetrans to a greater extent than geographical area in Ontario. Adults of P. penetrans from southern Ontario tended to be smaller than those reported elsewhere. The presence of three lip annules was consistent in the seven populations studied, although in some specimens one of the annules did not entirely encompass the head. Crenations around the tail tip of females of P. penetrans was common in the populations studied. Not all of the morphological characters were proportional in size to length of the females of P. penetrans.

Growth of Carrot and Tomato from Oxamyl-coated Seed and Control of Meloidogyne hapla.

Oxamyl was coated on carrot (Daucus carota L. cv. Spartan Fancy-80) and tomato (Lycopersicon esculentum Mill. cv. Glamour) seeds with a polymer sticker for the control of Meloidogyne hapla. The sticker diluted in water 1:1 delayed carrot seedling emergence. Oxamyl at 40 mg/ml in a 1:5 dilution of sticker lowered the rate of carrot seedling emergence until day 13 and plant growth until day 28. Oxamyl at 20 or 40 mg/ml in a 1:5 dilution of sticker on carrot seeds planted in M. hapla-infested muck soil resulted in fewer galled tap roots and fewer galls per root system 4 weeks after planting. Tap root lengths were greater than those of the control. Tomato seedling emergence was delayed and top and root weights were reduced, relative to the control, at 25 days by the sticker diluted 1:1 to 1:3. Oxamyl at 20 or 40 mg/ml in a 1:5 diluted sticker delayed tomato seedling emergence. Top weights of tomato seedlings from seeds coated with 20 mg/ml of oxamyl in a 1:5 diluted sticker planted in a silt loam were greater than control top weights at 4 and 6 weeks. Root weights were greater than those of the control only at 4 weeks. There were fewer galls per gram of root on seedlings from oxamyl-coated seeds and fewer juveniles per pot of soil, relative to the controls, only at 4 weeks.

Coating Soybean Seed with Oxamyl for Control of Heterodera glycines.

Oxamyl coated on soybean (Glycine max (L.) Merr. cv. Elgin) seeds in solutions of 20, 40, 80, and 160 mg/ml had no serious deleterious effects on seedling emergence and growth when planted in sterile soil. Seedling emergence on day 3 was less than that of the uncoated control, but by day 7 emergence was equal to, or greater than, the control. Shoot and root growth from seed coated with oxamyl in 40 and 80 mg/ml solutions was greater than that of the control. In soil infested with soybean cyst nematode, Heterodera glycines, shoot weight of soybean plants from seeds coated with oxamyl in 80 mg/ml solution was 11 and 9% greater at weeks 3 and 7, respectively, than from uncoated seeds. Numbers of juveniles (J3 and J4) and adults of H. glycines observed on the roots of plants from oxamyl-coated seeds were 83, 42, and 49% less at weeks 3, 5, and 7, respectively, than numbers on the roots of the untreated control. Numbers of J2 extracted from the roots of plants from oxamyl-coated seeds were 75% less at weeks 5 and 7 than those extracted from roots of uncoated seeds. The numbers of J2 extracted from the soil planted to oxamyl-coated seeds were 51 and 33% less at weeks 5 and 7, respectively, than from soil planted to uncoated seed.

Infectivity of Pratylenchus penetrans on Alfalfa.

The infectivity of Pratylenchus penetrans on alfalfa seedlings cv. Du Pulls was studied. The dense root-hair zone was the preferred zone of penetration by females, males, and third-stage larvae. A lesion initially appeared as a water-soaked area at the root surface, becoming yellow and elliptical as the nematode entered the cortex, with dark-brown cells later appearing in the centre as the nematode fed. At 20 C, females penetrated roots earlier, faster, and in greater numbers than either males or third-stage larvae. Females penetrated roots at temperatures from 5 to 35 C, with maximum penetration between 10 and 30 C, while males and third-stage larvae penetrated roots only between 10 and 30 C with maximum penetration a t 20 C. Penetration of roots by females, males, and third-stage larvae increased after storage of 5 C for 35 days, but decreased after storage of 140 days or more. Combinations of the three life stages in pairs neither enhanced nor inhibited penetration of roots by individual life stages; males were not attracted to females. Increasing inoculum density up to 20 nematodes/seedling did not affect penetration.

Efficacy of oxamyl coated on alfalfa seed with a polymer sticker in pratylenchus and meloidogyne infested soils.

A polymer sticker was used as a coating in which oxamyl was applied to seeds of alfalfa cultivar Saranac for the control of Pratylenchus penetrans and Meloidogyne hapla. The sticker, diluted 1:1 (sticker:water) to 1:5, delayed seedling emergence during the first 4 days after planting. By day 13, however, emergence from all sticker treatments was comparable to the control. Shoot growth of seedlings at day 21 was less than that of the control only from seeds coated with a 1:1 dilution; root growth and nodulation were not affected. Sticker-coated seeds absorbed 30-58% as much water in 3.5 hours as was absorbed by uncoated seeds. Oxamyl concentrations of 40-160 mg/ml in a 1:5 sticker : water mixture had no adverse affect on seedling emergence, growth, and nodulation over 3 weeks. Oxamyl at 160 mg/ml was more effective against P. penetrans than M. hapla. Growth of alfalfa in P. penetrans-infested soil was greater than that of the control in each sampling for 11 weeks. The reduction of number of P. penetrans in soil and roots moderated slowly over 11 weeks from 90% to 60%. Shoot and root growth of alfalfa from oxamyl-coated seed in M. hapla-infested soil were greater than those of the control for 7 and 11 weeks, respectively. The reduction in the number of M. hapla in the soil and roots changed from 80% at 7 weeks to 15% at 11 weeks.

Anhydrobiosis in Pratylenchus penetrans and Tylenchorhynchus n. sp. in Cultivated Soils Cropped to Winter Rye.

Anhydrobiotes of Pratylenchus penetrans were found in two cultivated soils sown to rye in southern Ontario during the growing season. Anhydrobiotes at the 0-2.5-cm depth were recovered from 9 and 6 of 11 samplings, respectively, of a Vineland silt loam and a Fox loamy sand during the dry summer of 1983. At the 2.5-15.0-cm depth, anhydrobiotes were recovered less frequently. In the summer of 1984, anhydrobiotes of P. penetrans were recovered once and anhydrobiotes of Tylenchorhynchus n. sp. twice in 11 samplings. The percentages of P. penetrans populations that were anhydrobiotes in 1983 and 1984 were closely related to soil moisture content and corresponding moisture tensions. Populations of P. penetrans were greatest in October in the lower soil depth, 2.5-15.0 cm; those of Tylenchorhynchus n. sp. were greater in the surface layer of soil, 0-2.5 cm, and peaked in August.

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.

Effect of Oxamyl Treatment of Potato Seed Pieces on Pratylenchus penetrans and Yield.

In the first 2 years of a 3-year (1987-89) microplot study, aqueous solutions of oxamyl (32 mg/ml) were applied to cut potato (Solanum tuberosum cv. Russet Burbank) tubers, grown in fine sandy loam infested with Pratylenchus penetrans. The seed-piece treatment alone and the seed-piece treatment followed by three foliar sprays generally reduced (P = 0.05) population densities of P. penetrans in the soil at midseason and in the soil and roots at harvest, compared to the control. In 1989, all seed pieces treated with oxamyl at 96 mg/ml or at 32, 64, and 96 mg/ml followed by a polymer sticker : water (1:4) dip failed to emerge. Only oxamyl at 64 mg/ml reduced (P = 0.05) midseason soil population densities of P. penetrans. A pre-plant soil treatment with 1,3-D reduced (P = 0.05) numbers of P. penetrans at planting each year and increased (P = 0.05) tuber yields in 1988 and 1989 compared to the control. In 1989, tuber yields from the sticker treatment and the oxamyl seed-piece treatment at 64 mg/ml were lower (P = 0.05) than those in the 1,3-D treatment and similar to those from the untreated control, possibly because of phytotoxicity. Oxamyl treatment of potato seed pieces to control P. penetrans does not appear practical for field production.

Growth of Potato and Control of Pratylenchus penetrans with Oxamyl-treated Seed Pieces in Greenhouse Studies.

Oxamyl was applied to both uncut and cut potato tubers in aqueous solutions of 1,000 to 32,000 mug/ml. Emergence in greenhouse pots was delayed for a day or more after soaking cut tuber pieces in 32,000 mug/ml. After 10 weeks plant growth was greater, relative to the control, when Pratylenchus penetrans-infested soil was planted with cut tubers soaked for 20 minutes in 32,000 mug/ml. Soaking for 40 minutes did not increase nematode control nor affect plant growth. Oxamyl applied to tubers at 1,000 mug/ml reduced the numbers of P. penetrans in the soil by 20% and in the roots by 35%; at 32,000 mug/ml, the numbers of P. penetrans in the soil were reduced by 73-86% and in the roots by 86-97%. The numbers of P. penetrans did not increase in the roots of plants developed from cut tubers soaked in 32,000 mug/ml over a period of 10 weeks, but numbers of lesion nematodes had begun to increase in the soil.

Control of Pratylenchus penetrans and Meloidogyne hapla and Yield Response of Alfalfa Due to Oxamyl Seed Treatments.

Alfalfa (Medicago sativa L. cv. Saranac) seed were soaked for 20 minutes in water, acetone, or methanol containing 10 or 50 mg/ml of oxamyl (Vydate L) or coated with a 2% aqueous cellulose solution containing the same amounts of oxamyl. Seed were analyzed for oxamyl by HPLC immediately after treatment and after 9 and 26 months of storage. Oxamyl content of alfalfa seed did not decline after 26 months of storage. The effects of seed treatment on growth of alfalfa and nematode control were examined using soils infested with Pratylenchus penetrans and Meloidogyne hapla. Germination was not affected by any of the seed treatments. Twenty-one days after sowing, the total growth of alfalfa seedlings grown from seed treated with 50 mg/ml of oxamyl in P. penetrans-infested soils had increased by 62% over controls. Nodulation per pot increased by as much as 267%, and the densities of P. penetrans per gram of root were reduced by as much as 73% compared to control plants. In M. hapla-infested soils, increases in plant growth (32%) and nodulation (71%) also occurred with oxamyl-treated seeds. Root gall reduction (86%) was also substantial due to oxamyl seed treatment.

Growth response of three vegetables to smooth- and crenate-tailed females of three species of pratylenchus.

The effect of morphological variants of females of Pratylenchus penetrans, P. neglectus, and P. crenatus on the growth of three vegetables was studied. Variants were characterized by having either a smooth or crenate tail terminus. Pea was inoculated with variants of P. penetrans, one female per seedling, and grown at light intensities ranging from 1,350 to 21,600 lux in a series of five experiments. Only crenate-tailed females of P. penetrans suppressed the growth of pea and only when pea was grown at 3,900 lux. Radish was inoculated with morphological variants of P. penetrans, P. neglectus, and P. crenatus, four females per seedling, and grown at 3,900 lux in two experiments. Again, truly creuate-tailed females of P. penetrans inhibited growth. The two variants of P. penetrans had a similar infectivity, greater than that of the other two species of Pratylenchus. Only crenate-tailed P. penetrans reproduced on radish. Onion was inoculated with variants of P. penetrans and P. crenatus, four females per seedling, and grown at 14 C at 12,900 lux. Again, only crenate-tailed P. penetrans inhibited growth. The variants of P. penetrans had a similar infectivity, greater than that of P. crenatus. Neither species reproduced on onion at low temperatures.

Biological Control of Meloidogyne hapla on Alfalfa and Tomato with the Fungus Meria coniospora.

This study was to determine whether Arthrobotrys flagrans, A. oligospora, and Meria coniospora would control the root-knot nematode Meloidogyne hapla on alfalfa and tomato. Alfalfa seeds were coated with a fungus-rye powder in 2% cellulose and were planted in infested soil. Three-week-old seedlings from seed treated with M. coniospora had 60% and 58% fewer galls in two experiments than did seedlings from untreated seeds. Numbers of J2 in the soil were not reduced. Plant growth did not improve. When seed of tomato were coated with M. coniospora and planted in M. hapla-infested soil, roots had 34% fewer galls and 47% fewer J2 in the soil at 28 days. After 56 days there was no reduction in J2 numbers. Plant growth did not improve. When roots of tomato transplants were dusted with M. coniospora fungus-rye powder or sprayed with a spore suspension before planting in M. hapla-infested soil, 42% and 35%, respectively, fewer galls developed in 28 days on treated roots than on roots not treated with fungus. The numbers of J2 extracted from roots or recovered from soil were not reduced, however, and plant growth did not improve.

Ultrastructural Pathology of Cells Affected by Pratylenchus penetrans in Alfalfa Roots.

Cortical parenchyma cells penetrated and fed upon by Pratylenchus penetrans for 48 hours contained only cytoplasmic debris. Proximal cells had an increase in tannin deposits, degenerated mitochondria, increased numbers of ribosomes, and no internal membrane structure. Often the endodermis was collapsed and contained massive tannin deposits on the inner cell wall and cell lumen. Similar observations were made in the stele, except tannin deposits were not as prominent. Multivesicnlate structures were observed both in the endodermis and in the stele.

Population development and reproduction of fungus- and bacterium feeding nematodes in the presence of insect growth regulators.

The insect growth regulators (IGRs), diflubenzuron and BAY SIR 8514, at 300 and 1,000 ppm a.i. in potato dextrose agar (PDA) inhibited the radial growth of the fungus Rhizoctonia solani host of Aphelenchus avenae. The IGRs had no effect on the growth of the bacterium Pseudomonas pseudoalcaligenes host of Acrobeloides nanus and Diplogaster iheritieri. At 59 ppm a.i., neither IGR inhibited the population development of A. nanus and D. iheritieri on P. pseudoalcaligenes; however, diflubenzuron stimulated the population development of D. iheritieri. At 300 ppm, both IGRs inhibited the population development of A. nanus and D. iheritieri; however, BAY SIR 8514 was more effective than diflubenzuron except on A. nanus L's. At 300 ppm, only BAY SIR 8514 affected the population development of A. avenae, except the L's. At 1,000 ppm, both IGRs inhibited development, except diflubenzuron for L and L's. Again, BAY SIR 8514 was more effective than diflubenzuron. With single females of A. nanus and D. iheritieri, both IGRs at 300 ppm reduced egg laying, inhibited embryonation, and slowed larval development.

Inhibition of Acetylcholinesterases from Aphelenchus avenae by Carbofuran and Fenamiphos.

Exposure to carbofuran and fenamiphos for 72 hours reduced the numbers of active Aphelenchus avenae in aqueous suspension by > 75%. When nematicides were removed, many A. avenae exposed to carbofuran resumed normal movement but A. avenae treated with fenamiphos did not recover. Acetylcholinesterase (AChE) activity was suppressed by > 95% in nematodes treated with carbofuran or fenamiphos. However, 48 hours after treated nematodes had been placed in water, AChE activity in carbofuran treated populations was 98% of the levels in control nematodes. Nematodes that had been treated with fenamiphos showed only slight AChE recovery. The antidotes, atropine sulfate and 2-PAM, were largely ineffective in counteracting the toxic effects of the nematicides.

Carbamate and Organophosphorus Nematicides: Acetylcholinesterase inhibition and Effects on Dispersal.

The sensitivities of acetylcholinesterases (ACHE) from the fungus-feeder Aphelenchus avenae and the plant-parasitic species Helicotylenchus dihystera and Pratylenchus penetrans and the housefly, Musca domestica, were compared using a radiometric assay which utilized H(3) acetylcholine as a substrate. Nematode ACHE were generally less sensitive to inhibition by organophosphorns and carbamate pesticides than were ACHE from the housefly. ACHE from the plant-parasitic species and A. avenae were generally similar in sensitivity. In soil, carbamates were more toxic than the organophosphorus pesticides to A. avenae. All pesticides tested affected nematode movement, but fenamiphos was more inhibitory than others. The effects on dispersal of nematodes may be an important mechanism in control by some nematicides.

Sensitivity of Acetylcholinesterases from Aphelenchus avenae to Organophosphorous and Carbamate Pesticides.

The sensitivities of acetylcholinesterases (ACHE) from the nematode Aphelenchus avenae and the house fly Musca domestica to various pesticides were compared using a colorimetric assay. ACHE from A. avenae were generally less sensitive than ACHE from M. domestica to inhibition by organophosphorous and carbamate pesticides. Carbamates were somewhat more inhibiting than organophosphorous pesticides to nematode ACHE. In vivo tests with concentrations of various pesticides up to 500 ppm in sand caused less than 100% mortality of nematodes.