Influence of Low Temperature on Rate of Development of Meloidogyne incognita and M. hapla Larvae

Development of Meloidogyne incognita and M. hapla larvae in clover roots was studied at 20, 16, 12, and 8 C in growth chambers and in the field from fall through spring, in North Carolina. Larvae of both species invaded roots and developed at 20, 16, and 12 C, but not at 8 C. The time necessary to complete the larval stages at each temperature was determined. The minimal temperature for development of M. incognita larvae was 10.08 C and 8.8 C for M. hapla larvae. In the field, soil temperature at 10 cm deep was favorable for development of larvae until the end of November, and again from February on. All stages of the nematodes survived freezing temperatures in the roots. Reproduction of both species was evident in March or Apri1 after inoculation and accumulation of 8,500 to 11,250 degree-hours.     

Relationships of Initial Population Densities of Meloidogyne incognita and M. hapla to Yield of Tomato

Microplots 80 × 100 cm, infested with varying initial population densities (P_i) of Meloidogyne incognita or M. hapla, were planted to tomato at two locations. Experiments were conducted in a sandy loam soil at Fletcher, N. C. (mountains) where the mean temperature for May to September is ca 20.7 C, and in a loamy saml at Clayton, N. C. (coastal plain) where the mean temperature for May to Septemher is ca 24.8 C. In these experimentally infested plots, M. incognita and M. hapla caused maximunt yield losses of 20-30%, at lhe mountain site with Pi of 0-12,500 eggs and larvae/500 cm³ of soil. In the coaslal plain, M. incognita suppressed yields up to 85%, and M. hapla suppressed yields up to 50% in comparison with the noninfested control. A part of the high losses at this site apparently was due to M. incognita predisposing tomato to the early blight fungus. In a second experintent, in which a nematicide was used to obtain a range of P_is (with P_i as high as 25,000/50 cm³ of soil) at Fletcher, losses due to M. incognita were as great as 50%, but similar densities of M. hapla suppressed yields by only 10-25%. Approximate threshold densities for both species ranged from 500 to 1,000 larvae and eggs (higher for surviving larvae) for the mountain site, whereas nutnbers as low as 20 larvae/500 cm³ of soil of either species caused signiticant damage in the coastal plain. Chemical soil treatments proved useful in obtaining various initial population densities; however, problems were encountered in measuring effective inoculum after such treatments, especially in the heavier soil.     

Influence of Low Temperature on Development of Meloidogyne incognita and M. hapla Eggs in Egg Masses

Egg masses of Meloidogyne incognita and M. hapla were placed in soil at 10, 12, 16, and 20 C. At regular intervals, eggs from samples of egg masses were released from the gelatinous matrices and their developmental stages recorded. The number of days necessary to complete each stage from gastrulation to hatch is given for each temperature. The minimal temperature threshold for the development of eggs was computed by linear regression to be 8.26 C for M. incognita and 6.74 C for M. hapla.     

Retention of Resistance to Meloidogyne incognita in Lycopersicon Genotypes at High Soil Temperature

Lycopersicon glandulosum and L. peruvianum clones and L. esculentum cultivars ''VFN8'' (resistant) and ''Rutgers'' (susceptible) were tested for their resistance to Meloidogyne incognita (race l) at soil temperatures of 25 and 32 C. L. esculentum cv. VFN8 and L. peruvianum Acc. No. 128657, both of which possess the Mi gene, were resistant at 25 C but were susceptible at 32 C. L. glandulosum Acc. No. 126443 and L. peruvianum Acc. No. 270435, with combined resistance to M. hapla and M. incognita, and L. peruvianum Acc. Nos. 129152 and LA2157, with resistance to M. incognita, were highly resistant at both temperatures. In a second experiment three of these accessions under heat stress simulated by 32 C ambient and soil temperature retained a high level of resistance. Two clones of L. glandulosum Acc. No. 126440, with resistance to M. hapla, were moderately susceptible to M. incognita at 25 and highly susceptible at 32 C. M. incognita produced significantly (P = 0.01) more eggs on L. esculentum cv. Rutgers at 32 than at 25 C. This study supports the existence of genes other than the Mi gene that confer resistance to M. incognita and are functional at high soil temperatures.     

Interrelationships of Meloidogyne Species with Flue-cured Tobacco

Microplot and field experiments were conducted to determine relationships of population densities of Meloidogyne spp. to performance of flue-cured tobacco. A 3-yr microplot study of these interactions involved varying initial nematode numbers (P_i).and use of ethoprop to re-establish ranges of nematode densities. Field experiments included various nematicides at different locations. Regression analyses of microplot data from a loamy sand showed that cured-leaf yield losses on ''Coker 319'' for each 10-fold increase in P_i were as follows: M. javanica and M. arenaria—-13-19%; M. incognita—5-10%; M. hapla—3.4-5%; and 3% for M. incognita on resistant ''Speight G-28'' tobacco. A P_i of 750 eggs and larvae/500 cm³ of soil of all species except M. hapla caused a significant yield loss; only large numbers of M. hapla effected a loss. M. arenaria was the most tolerant species to ethoprop. Root-gall indices for microplot and most field-nematicide tests also were correlated negatively with yield. Relationships of P_i(s) and necrosis indices to yield were best characterized by linear regression models, whereas midseason numbers of eggs plus larvae (P_m) and sometimes gall indices vs. yield were better characterized by quadratic models. The relation of field P_m and yield was also adequately described by the Seinhorst model. Degrees of root galling, root necrosis, yield losses, and basic rates of reproduction on tobacco generally increased from M. hapla to M. incognita to M. arenaria to M. javanica.     

Influence of Photoperiod and Temperature on Migrations of Meloidogyne Juveniles

Photoperiod influences the migration of M. incognita juveniles toward tomato roots. Approximately 33% migrated vertically 20 cm in 7 days to roots when 12 h dark were alternated with 12 h light. Only 7% migrated when light was constant for 24 h. Vertical migration of M. incognita juveniles was studied at 14, 16, 18, 20, and 22 C. The migration of M. incognita juveniles begins at about 18 C and reaches its maximum at 22 C. The migration of M. hapla and M. incognita juveniles were compared at 14, 18, and 22 C. Juveniles of M. hapla were able to migrate at a lower temperature than those of M. incognita. With M. hapla, there was no significant difference in migration between 18 and 22 C.     

Application of Isoelectric Focusing to the Taxonomic Identification of Meloidogyne spp.

Meloidogyne incognita, M. arenaria, M. hapla, and M. javanica were distinguishable from each other by isoelectric focusing (IEF) of nematode egg proteins. Proteins extracted from larvae and adults of Hoplolaimus columbus and from eggs of Heterodera glycines had distinctive profiles, also. Protein profiles from eggs, preparasitic larvae and egg-laying adults of M. incognita showed differences. It was necessary to compare samples run at the same time to ensure reliability.     

Effects of Temperature and Root Leachates on Embryogenic Development and Hatching of Meloidogyne chitwoodi and M. hapla

At 20 C the duration of the embryogenic development of Meloiclogyne chitwoodi and M. hapla was about 20 days. At 10 C the embryogenic development was 82-84 days for M. chitwoodi and 95-97 days for M. hapla. The effect of distilled water and root leachates of potato cv. Russet Burbank, tomato cv. Columbian, and wheat cv. Hyslop on the hatching of eggs of the two root-knot nematode species was investigated at 4, 7, 10, 15, 20, and 25 C (± 1 C). Cumulative egg taatch was no greater in root leachates titan in distilled water, but temperature did significantly affect egg hatch (P = 0.05). Less than 1% of the eggs of both nematode species hatched at 4 C. The percent cumulative hatch at 10 C was significantly less (P = 0.05) than at higher temperatures for both nematodes and significantly more (P = 0.05) M. chitwoodi eggs hatched than did M. hapla eggs. At 15 G the percent cumulative hatch of both species was significantly lower (P = 0.05) than that at 20 and 25 C. The percent cumulative egg hatch of two species did not differ at 25 C, but was higher (P = 0.05) at 25 C than at 20 C. At 7 C the emergence of M. chitwoodi juveniles was about seven times (P = 0.01) greater than that of M. hapla in distilled water.     

Adaptations to the Arctic: low-temperature development and cold tolerance in the free-living stages of a parasitic nematode from Svalbard

For nematodes with a direct life cycle, transmission is highly dependent on temperature-related development and survival of the free-living stages. Therefore, in the Arctic, where the winter lasts from October to May, nematode transmission is expected to be focused in the short summer season, yet there is strong evidence that as well as focussing egg output during winter months, the nematode parasite, Marshallagia marshalli, infects Svalbard reindeer during the Arctic winter when temperatures are persistently below freezing. To investigate the potential for development and survival of eggs and infective third-stage larvae in winter and therefore the possibility of for winter transmission, we ran a series of low-temperature laboratory experiments. These provide five key insights into the transmission and survival of the free-living stages of M. marshalli: (1) eggs hatched at temperatures as low as 2 °C, but not below 0 °C, (2) eggs were viable and developed after being exposed to sub-zero temperatures for up to 28 months, (3) infective-stage larvae survived for up to 80 days at 5 °C, (4) infective-stage larvae could survive rapid exposure to temperatures below ?30 °C, and (5) desiccation resistance may be important for long-term larval survival at low temperatures. Together, these results indicate that eggs deposited during the winter are highly tolerant of prevailing environmental conditions and have the potential for rapid development with the onset of spring. It is therefore likely that the parasite remains in the egg stage in the faeces during the winter of deposition, hatch and develop into the infective larval stage in the summer, remaining viable on the tundra until the reindeer host returns to the winter feeding grounds the following winter.     

Penetration and Development of Meloidogyne hapla in Resistant and Susceptible Alfalfa Under Differing Temperatures

Studies were conducted to examine under differing temperatures (12, 16, 20, 24, 28, and 32 C) the penetration anti development of Meloidogyne hapla in resistant lines ''298'' and ''Nev. Syn XX'', and susceptible ''Lahontan'' and ''Ranger'' hardy-type alfalfas. The results indicated that resistance to M. hapla was similar to that previously described for M. incognita in nonhardy alfalfa. Although initial penetration in resistant seedlings was similar to that of susceptible seedlings, nematode larvae failed to establish and develop in root tissues and nematode numbers subsequently declined. In susceptible seedlings, nematode development proceeded rapidly, and egg production began after 5 weeks. Temperature had little influence on the nematode development except to slow the response at the lower temperatures. Other studies were conducted to verify a previously reported immune (no penetration) reaction to M. hapla by the ''Vernal'' selection ''M-4''. When compared to the resistant (penetration without nematode development) Vernal selection ''M-9'' under differing temperatures (20, 24, 28, and 32 C), each selection was equally penetrated by M. hapla but at a lower level than in susceptible Ranger cuttings. Generally, no root galling was observed in either M-4 or M-9; however, very slight galling was found 35 days after inoculation on about 50% of these cuttings when grown at 32 C.     

Role of Nematodes,Nematicides, and Crop Rotation on the Productivity and Quality of Potato,Sweet Potato,Peanut, and Grain Sorghum

The objective of this experiment was to determine the effects of fenamiphos 15G and short-cycle potato (PO)-sweet potato (SP) grown continuously and in rotation with peanut (PE)-grain sorghum (GS) on yield, crop quality, and mixed nematode population densities of Meloidogyne arenaria, M. hapla, M. incognita, and Mesocriconema ornatum. Greater root-gall indices and damage by M. hapla and M. incognita occurred on potato than other crops. Most crop yields were higher and root-gall indices lower from fenamiphos-treated plots than untreated plots. The total yield of potato in the PO-SP and PO-SP-PE-GS sequences increased from 1983 to 1985 in plots infested with M. hapla or M. arenaria and M. incognita in combination and decreased in 1986 to 1987 when root-knot nematode populations shifted to M. incognita. The total yields of sweet potato in the PO-SP-PE-GS sequence were similar in 1983 and 1985, and declined each year in the PO-SP sequence as a consequence of M. incognita population density increase in the soil. Yield of peanut from soil infested with M. hapla increased 82% in fenamiphos-treated plots compared to untreated plots. Fenamiphos treatment increased yield of grain sorghum from 5% to 45% over untreated controls. The declining yields of potato and sweet potato observed with both the PO-SP and PO-SP-PE-GS sequences indicate that these crop systems should not be used longer than 3 years in soil infested with M. incognita, M. arenaria, or M. hapla. Under these conditions, these two cropping systems promote a population shift in favor of M. incognita, which is more damaging to potato and sweet potato than M. arenaria and M. hapla.     

Effect of soil temperature and moisture on survival of eggs and first-instar larvae of Delia radicum

Edaphic factors such as soil temperature and moisture influence soil-dwelling insects, whose most vulnerable stages typically are eggs and young larvae. In this study, the survival of eggs and first-instar larvae of the cabbage maggot, Delia radicum L., was measured under laboratory conditions after exposure to a range of soil temperatures and moistures. When eggs were exposed to constant temperature (20-29°C) and humidity (5-200% [wt:wt]), temperature had no significant effect on survival, whereas humidity <25% [wt:wt] caused egg mortality. The gradual exposure of eggs to high temperatures resulted in low mortality below 33°C, but <5% of eggs survived at 40°C. When first-instar larvae were exposed to constant temperature (17-29°C) and humidity (5-100% [wt:wt]), both factors as well as their interaction had a significant effect on larval survival, which was nil at 5% (wt:wt) for all temperatures but increased from 21.9 to 42.8% at 17°C and from 34.1 to 55.0% at 29°C, for soil moisture contents of 15% and 100% (wt:wt), respectively. Eggs of D. radicum are resistant to low soil moisture and high temperature conditions. Larval survival tends to increase with an increase in soil temperature and moisture. It is suggested that soil temperature be integrated into insect development simulation models instead of air temperature, to build more effective models for cabbage maggot management.     

Nature of Sweet Potato Resistance to Meloidogyne incognita and the Effects of Temperature on Parasitism

Penetration, rate of development, and total population of Meloidogyne incognita in roots of susceptible ''Allgold'' and resistant ''Nemagold'' sweet potatoes increased with temperature 24-32 C. Rate of larval penetration in ''Allgold'' was significantly higher than in ''Nemagold'' after 48 hr of root exposure at 24, 28, and 32 C. At 24, 28, and 32 C (16 hr) day and 20 C (8 hr) night temperature the life cycle of M. incognita required 42, 32, and 28 days in ''Allgold'', and 44, 33, and 31 days in ''Nemagold''; mature females in the first generation were 40, 40, 40, and 10, 22, 20 respectively. The correlation between the length of time roots were allowed to grow in the soil prior to inoculation and number of larvae recovered from the roots after inoculation was positive for ''Allgold'' and negative for ''Nemagold''. Therefore, a root exudate repellent to M. incognita larvae is proposed as a hypothetical basis for resistance to M. incognita in sweet potatoes.     

Histopathology of Selected cultivars of Tobacco infected with Meloidogyne species

Rates of nematode penetration and the histopathology of root infections in fluecured tobacco cultivars ''McNair-944,'' ''Speight G-28,'' and ''NC-89'' with either Meloidogyne arenaria, M. incognita, M. hapla, or M. javanica were investigated. Penetration of root tips by juveniles of all species into the M. incognita-resistant NC-89 and G-28 was much less than that on the susceptible McNair-944. Few juveniles of M. incognita were detected in resistant cultivars 7 and 14 days after inoculation. Infection sites exhibited some cavities and extensive necrotic tissue at 14 days; less necrotic tissue and no intact nematodes were observed 35 days after inoculation. Although some females of M. arenaria reached maturity and produced eggs, considerable necrosis was induced in the resistant cultivars. Meloidogyne hapla and M. javanica developed on all cultivars, but there was necrotic tissue at some infection sites in the resistant cultivars. The occurrence of single multistructured nuclei in the syncytia of most M. hapla infections differed from the numerous small nuclei found in syncytia caused by the other three species.     

In vitro storage of unfertilized eggs of the Eurasian perch and its effect on egg viability rates and the occurrence of larval malformations

Ova ageing is the most important factor affecting fish egg quality after ovulation. Long-term storage of fish ova, using cryopreservation and vitrification techniques, has been unsuccessful to date. Instead, short-term in vitro ova storage has been used successfully and optimized in some cultured fish species. In vitro ova storage can drastically improve mass production of larvae and juveniles in the hatcheries by providing the possibility of the synchronous artificial fertilization for different females. To study how long unfertilized eggs of Eurasian perch (Perca fluviatilis L.) can retain their fertilizing ability after stripping, eggs were stored at temperatures of 4°C, 8°C and 12°C for 72 h post-stripping (HPS). The stored eggs of four female perch were separately fertilized at 0 h (i.e. control eggs fertilized before storage) and at 6-hour intervals during the experimental period of 72 h. The embryos reaching the eyed-egg and hatched-larvae stages, eyed-egg mortality and larval malformation rates were recorded as indices of egg quality. The results indicated that the maximum eyed eggs and hatched larvae (86% and 63%, respectively) were observed for eggs fertilized immediately after stripping, whereas the storage of the eggs at 4°C for 48 HPS decreased the eyed-egg and hatched-larvae rates to 46% and 17%, respectively. The use of a higher storage temperature resulted in a more rapid decrease in egg viability: eyed-egg and hatched-larvae rates of 23% and 9%, respectively, were obtained after 48 HPS storage at 8°C and 2% and 1% for eggs stored at 12°C. Eyed-egg mortality and larval malformation rates were not significantly affected by post-stripping ova ageing for at least up to 36 h. Thereafter, both values increased significantly and were measured to be the highest in the most aged ova. The present study demonstrated that stripped Eurasian perch eggs can be stored for at least 12 h at 4°C to 12°C without a significant reduction in their quality.     

Interrelationship of Meloidogyne hapla and Ditylenchus dipsaci on Resistant and Susceptible Alfalfa

Simultaneous inoculations of alfalfa with Meloidogyne hapla larvae and Ditylenchus dipsaci at 16, 20, 24, and 28 C did not depress penetration of either nematode in ''Nev Syn XX'' -a selection resistant to M. hapla and D. dipsaci, ''Vernal 298'' -a selection resistant to M. hapla and susceptible to D. dipsaci, ''Lahontan'' -a cultivar resistant to D. dipsaci and susceptible to M. hapla, and ''Ranger'' -a cultivar susceptible to both M. hapla and D, dipsaci. Infection with D. dipsaci depressed growth of susceptible ''Vernal 298'' and ''Ranger'' at all soil temperatures, except for ''Vernal 298'' at 16 C. Infection with M. hapla alone did not depress growth of any of the alfalfas. A combination of M. hapla and D. dipsaci resulted in a synergistic weight depression on ''Ranger'' at all soil temperatures. Inoculation of the four alfalfas with D. dipsaci 2, 4, 6, and 8 wk before inoculation with M. hapla at 16, 20, 24, and 28 C did not influence the resistance or susceptibility of ''Nev Syn XX,'' ''Lahontan,'' or ''Ranger.'' However, galling of ''Vernal 298'' by M. hapla was affected by soil temperature, plant age, and inoculation with D. dipsaci.     

Development ofDiatraea Saccharalis [Lep.: Pyralidae] at constant temperatures

At constant temperatures between 15.6 and 32°C the incubation time of eggs ofDiatraea saccharalis (F.) was reduced by each increase in temperature. At 34°C the time decreased. Highest (98.6%) and lowest (9.9%) egg hatch occurred at 26 and 34°C, respectively. Larvae completed development at temperatures ranging from 22 to 34°C; however, only 4.4% of the larvae pupated at 34°C. Duration of the larval stage at 30°C (♂=18.1 days; ♀=19.1 days) was ca. 14 days shorter than at 22°C. Maximum rate of development in the pupal stage occurred at 28°C (ca. 6.8 days), and a higher temperature increased developmental time and mortality. Adult longevity and egg production generally were reduced with increasing temperatures and egg production was highest at 24°C (729.8 eggs/ moth). As many as 7 larval stages occurred; but most larvae completed development in 5 stages, and none completed development in less than 5 stages. The female larval stage was ca. 1 day longer than that of males, and this difference occurred primarily in the 5th stage.     

Damage Threshold of Meloidogyne hapla to Lettuce in Organic Soil

Lettuce was seeded in pots in the greenhouse and in field microplots in 1991 and 1992. Pots and microplots were filled with untreated or fumigated organic soil infested with Meloidogyne hapla at seven initial population densities (Pi) (0 to 32 eggs/cm³ soil). Lettuce weight, severity of root galling, and number of eggs per root system (Pf) were determined after 8 weeks. At the highest Pi, M. hapla caused yield losses up to 64% in the microplots and plant death in the greenhouse tests. The Seinhorst equation was used to describe the relation between lettuce weight and Pi (r² = 0.73 - 0.98) and to calculate the damage threshold density (T). Values of T were 7 and 8 eggs/cm³ soil in the greenhouse tests of 1991 and 1992, respectively. In the microplot tests, T was 1 egg/cm³ soil in 1991 and 2 eggs/cm³ soil in 1992. The damage threshold was the same in untreated and fumigated soils. At low Pi, root galling was more severe in the pots than in the microplots. Pf increased with increasing Pi of M. hapla in both tests, but declined at Pi > T in the greenhouse tests. The reproduction rate (Pf/Pi) of M. hapla was highest at the lowest Pi.     

Hirsutella rhossiliensisand Verticillium chlamydosporium as Biocontrol Agents of the Root-knot Nematode Meloidogyne hapla on Lettuce

Hirsutella rhossiliensis and Verticillium chlamydosporium infected second-stage juveniles (J2) and eggs of Meloidogyne hapla, respectively, in petri dishes and in organic soil in pots planted to lettuce in the greenhouse. In vitro, H. rhossiliensis produced 78 to 124 spores/infected J2 of M. hapla. The number of J2 in roots of lettuce seedlings decreased exponentially with increasing numbers of vegetative colonies of H. rhossiliensis in the soil. At an infestation of 8 M. hapla eggs/cm³ soil, 1.9 colonies of H. rhossiliensis/cm³ soil were needed for a 50% decrease in J2 penetration of lettuce roots. Egg-mass colonization with V. chlamydosporium varied from 16% to 43% when soil was infested with 8 M. hapla eggs and treated with 5,000 or 10,000 chlamydospores of V. chlamydosporium/cm³ soil. This treatment resulted in fewer J2 entering roots of bioassay lettuce seedlings planted in the infested soils after harvesting the first lettuce plants 7 weeks after infestation with M. hapla. Hirsutella rhossiliensis (0 to 4.3 colonies/cm3 soil), V. chlamydosporium (500 to 10,000 chlamydospores/cm3 soil), or their combination, added to organic soils with 8 M. hapla eggs/cm³ soil, generally did not affect lettuce weight, root galling, or egg production of M. hapla. However, when lettuce was replanted in a mix of infested and uninfested soil (1:3 and 1:7, v:v), egg production was lower in soils with V. chlamydosporium than in soils without the fungus. Both fungi have potential to reduce the M. hapla population, but at densities below 8 eggs/cm³ soil.     

Effect of Soil Temperature on Reproduction of Meloidogyne chitwoodi and M. hapla Alone and in Combination on Potato and M. chitwoodi on Rotation Plants

Meloidogyne chitwoodi developed and reproduced more rapidly than M. hapla in potato roots at 15, 20, or 25 C when both species of nematodes were inoculated simultaneously at 250 or 1,000 juveniles of each. At 30 C significantly more M. hapla than M. chitwoodi females were found at the lower inoculum level after 41 days. More M. chitwoodi than M. hapla juveniles were extracted from soil at 15, 20, and 25 C, but only at the lower inoculum level at 30 C. Potato was considered a more suitable host for M. chitwoodi than M. hapla because of M. chitwoodi''s greater reproduction at 15, 20, and 25 C. Corn and wheat cultivars tested supported M. chitwoodi reproduction at temperatures of 10, 15, 20, and 25 C, but fewest eggs were produced on these plants at 20 C. Temperatures of 10 to 25 C had little influence on the low reproduction of M. chitwoodi on four alfalfa cultivars. M. chitwoodi reproduced on the alfalfa entry Mn PL9HF.