Optimum Initial Inoculum Levels for Evaluation of Resistance in Tomato to Meloidogyne spp. at Two Different Soil Temperatures

The effects of Meloidogyne incognita or M. javanica at five initial inoculum levels of 20, 100, 200, 1,000, and 2,000 eggs and infective juveniles per seedling on ''Floradade,'' ''Nemarex,'' ''Patriot,'' and ''PI 129149-2(sib)-5'' tomatoes maintained at 25 or 32.5 C were studied. The number of egg masses on roots of the susceptible cultivar Floradade was similar for both species of root-knot nematodes at either 2.5 or 32.5 C soil temperatures. At 25 C, very low numbers of egg masses were produced by both species of root-knot nematodes on Nematex, Patriot, and Lycopersicon peruvianum PI 129149-2(sib)-5. At 32.5 C, the best inoculum level for assessing resistance in these tomato genotypes was 200 eggs and infective juveniles per seedling. With 28 days of incubation, this temperature and inoculum level produced quantitative differences in resistance for both species of Meloidogyne.     

An In Vitro Test for Temperature Sensitivity and Resistance to Meloidogyne incognita in Tomato

An in vitro root explant tissue culture technique is described for determining susceptibility of tomato (Lycopersicon esculentum Mill.) breeding lines and cultivars to the root-knot nematode Meloidogyne incognita. Root explants were taken from 2-day-old seedlings cultured for 30 days at 28 C on Gamborg''s B-5 medium with or without nematode inoculum. The remaining portion of the root and stem from the excised root explants was transferred to soil in pots and grown to maturity in the greenhouse. In vitro root explants were evaluated for growth and occurrence of juveniles, adults, and egg masses. The regenerated plants were used to produce more seed, The proposed technique is simple, reliable, and adapted to routine screening of large numbers of F₁ and F₂ samples, and it utilizes less space than tests performed on intact plants in the greenhouse or growth chamber. Evidence is presented also on the breakdown of resistance to M. incognita under high temperature stress using this in vitro root explant technique.     

Pathological Interaction of a Combination of Heterodera schachtii and Meloidogyne hapla on Tomato

Increased culturing of a tomato population of Heterodera schachtii (UT1C) on tomato for 480 days (eight inoculation periods of 60 days each) significantly increased virulence to ''Stone Improved'' tomato. A synergistic relationship existed between Meloidogyne hapla and H. schaehtii on tomato. A combination of H. schachtii (UTIC) and M. hapla significantly reduced tomato root weights by 65, 64, and 61% below root weights of untreated controls, and single inoculations of M. hapla and H. schachtii, respectively. This corresponded to root reductions of 42, 44, and 46% from a combination of H. schachtii (UT1B) and M. hapla. Antagonism existed between H. schachtii and M. hapla with regard to infection courts and feeding sites. The root-knot galling index dropped from 6.0 with a single inoculation of M. hapla to 4.3 and 3.3 with combined inoculations of M. hapla plus UT1B and M. hapla plus UTIC cyst nematode populations. The pathological virulence of H. schachtii to sugarbeet was not lost by extended culturing on tomato; there were no differences in penetration, maturation, and reproduction between sugarbeet populations continually cultured on sugarbeet and the population continually cultured on tomato.     

Mass Culture of Subanguina picridis and Its Bioherbicidal Efficacy on Acroptilon repens

A Russian knapweed (Acroptilon repens) shoot culture system, initiated from shoot tip culture, was used to generate a source of host plant tissue for the rearing of the nematode Subanguina picridis, a biocontrol agent for Russian knapweed. Young shoots growing on solid B5G medium in petri dishes developed galls on leaves, petioles, and shoot tips 7 days after release of 50 nematodes onto the surface of the medium. After 3 months of culturing, each petri dish yielded 7,000-10,000 nematodes. In vitro cultured Subanguina picridis were virulent on greenhouse-grown Russian knapweed plants. Galls were first found on seedlings 12 days after infestation; after 2 months, 90% of seedlings were galled on leaves, petioles, and shoot tips, with 1-6 galls per seedling. Three months after shoot emergence, 64% of vegetative shoots originating from root segments were also galled by the cultured nematodes. Similarly, vegetatively regenerated shoots of Russian knapweed were also susceptible to infestation by cultured nematodes.     

Evidence for a Dosage Effect of the Mi gene on Partially Virulent Isolates of Meloidogyne javanica

The reproduction of single egg-mass isolates of Meloidogyne javanica from Crete that differed in virulence were compared on tomato (Lycopersicon esculentum) genotypes homozygous or heterozygous for the Mi gene. The reproduction of three isolates with partial virulence was much greater on tomato genotypes heterozygous for the Mi gene (cultivars Scala, Bermuda, and 7353) than on two homozygous genotypes (F8 inbred lines derived from Scala). The reproduction of a highly virulent isolate on the homozygous and heterozygous genotypes was similar to that on a susceptible cultivar. These results pose questions regarding the nature of partial virulence and indicate a quantitative effect of the Mi gene in relation to such virulence.     

A Pathotype System to Describe Intraspecific Variation in Pathogenicity of Meloidogyne chitwoodi

Tests of eight Dutch Meloidogyne chitwoodi isolates to the differential set for host races 1 and 2 in M. chitwoodi provided no evidence for the existence of host race 2 in the Netherlands. The data showed deviations from expected reactions on the differential hosts, which raised doubts of the usefulness of the host race classification in M. chitwoodi. The term ''''pathotype'''' is proposed for groups of isolates of one Meloidogyne sp. that exhibit the same level of pathogenicity on genotypes of one host species. We recommend that the pathotype classification be applied in pathogen-host relationships when several genotypes of a Meloidogyne sp. are tested on several genotypes of one host species. Three pathotypes of M. chitwoodi were identified on Solanum bulbocastanum, suggesting at least two different genetic factors for virulence and resistance in the pathogen and the host species, respectively. The occurrence of several virulence factors in M. chitwoodi will complicate the successful application of resistance factors from S. bulbocastanum for developing resistant potato cultivars.     

Parasitism of Hoplolaimus galeatus on Diploid and Polyploid St. Augustine grasses

''Floratam'' and ''FX-313'' St. Augusfinegrasses (Stenotaphrum secundatum) were compared in a time-course experiment for host suitability and susceptibility to the lance nematode, Hoplolaimus galeatus. Nematode densities were determined in the soil and acid-fuchsin stained roots 42, 84, 126, 168, and 210 days after pots containing 230 cm³ of autoclaved native Margate fine sand/pot were infested with 104 ± 9 nematodes and maintained at 25 ± 2 C in the laboratory. ''FX-313'' was a more suitable host for H. galeatus. Numbers of H. galeatus reached a maximum at 210 days after inoculation, with 5,550 and 4,120 nematodes (adults plus juveniles)/pot for ''FX-313'' and ''Floratam,'' respectively. Root and shoot dry weights of both grasses were not affected by H. galeatus throughout the experiment. Three polyploid, 2n = 30 to 32 (''Floratam,'' ''FX-10,'' and ''Bitterblue'') and three diploid, 2n = 18 (''FX-313,'' ''Florida Common,'' and ''Seville'') S. secundatum genotypes were inoculated with H. galeatus (99 ± 9/pot) and compared with uninoculated controls 210 days after inoculation. St. Augustinegrass genotypes differed as hosts of H. galeatus. ''FX-313'' and ''Florida Common'' represented the high and low extremes, respectively, for nematode reproduction (9,750 and 5,490 nematodes/pot or 4,239 and 2,387 nematodes/100 cm³ of soil). However, differences in root and shoot growth were not detected 210 days after inoculation with H. galeatus.     

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.     

Relationship between Meloidogyne incognita and Rotylenchulus reniformis as Influenced by Soybean Genotype

The effect of soybean genotype on competition between Meloidogyne incognita race 2 (Mi) and Rotylenchulus reniformis (Rr) was evaluated in greenhouse and microplot replacement series experiments. Soil in pots containing seedlings of ''Davis'' (susceptible to Mi) or ''Buckshot 66'' (resistant to Mi) was infested with 1,000 vermiform individuals in the following Mi:Rr ratios: 0:0, 100:0, 75:25, 50:50, 25:75, or 0:100. After 91 days, the relative nematode yields (number of nematodes in mixed culture divided by the number in nonmixed culture) of each species were calculated based on soil and root nematode populations expressed as nematodes per gram of dry root tissue. To define the relationship between the two species, calculated relative nematode yields were compared with a theoretical noncompetition model using lack-of-fit regression. In the greenhouse, Mi populations on ''Davis'' were stimulated in the presence of Rr. In microplots, low Mi and Rr population densities likely resulted from severe galling and destruction of feeder roots that probably occurred early in the season. Enhanced susceptibility to Mi was not observed on ''Buckshot 66'', which remained resistant to Mi even when colonized by Rr. Host resistance is a key factor in determining the nature of the relationship between Mi and Rr.     

In Vitro Culture and Feeding Behavior of Belonolaimus longicaudatus on Excised Zea mays Roots

A greenhouse population of the sting nematode, Belonolaimus longicaudatus, obtained from an infested golf course in California''s Coachella Valley, was surface-decontaminated and cuhured on excised roots of Zea mays supported by Gamborg''s B5 medium. At 26-27 °C the females laid eggs, and newly emerged juveniles of the second generation completed three molts within 29 days after egg deposition. Sixty days after inoculation with 60 females and 40 males, an average of 529 nematodes and 83 eggs were recovered from the culture. The feeding process consisted of probing, stylet penetration, ingestion, and stylet retraction. Feeding seemed to be necessary before egg deposition or molting occurred. The sting nematode was observed feeding exclusively as an ectoparasite and preferably at the region of cell division and elongation. Vigorous feeding by many nematodes usually caused discoloration of root tips and termination of growth.     

Penetration of Crotalaria juncea,Dolichos lablab,and Sesamum indicum Roots by Meloidogyne javanica

Penetration of Crotalaria juncea (PI 207657 and cv. Tropic Sun) Dolichos lablab cv. Highworth, and Sesamum indicum by juveniles (J2) of Meloidogyne javanica was assessed to investigate the mechanism by which these plants may reduce nematode numbers in the field. Growth chamber experiments were conducted at 25 C, with vials containing 90 g sand infested with 450 J2; tomato (UC 204 C) was included as a susceptible host. Fifteen days after inoculation, roots were stained and the nematodes within stained roots were counted. Both C. juncea lines were highly resistant to penetration, as they contained significantly fewer nematodes per cm of root and per root system than the other plants. Although containing more nematodes per cm of root than C. juncea, S. indicum and D. lablab had significantly fewer nematodes per root system and per cm of root than tomato. Roots were significantly longer in the plants with the lowest nematode penetration. Although C. juncea, D. lablab, and S. indicum may have potential utility as cover or rotation crops in soil infested with M. javanica, further quantitative information on the reproduction of M. javanica and other nematodes in these plants is needed.     

不同番茄材料对B型烟粉虱个体发育和繁殖能力的影响

以甘蓝寄主上连续繁殖多代后的B型烟粉虱为对象,对其在8种番茄材料(4个栽培番茄、3个多毛番茄和1个醋栗番茄)上的产卵量、体型大小、发育历期、存活率以及第2代成虫的产卵量和寿命等生物学参数进行观察.自然情况下(10:00-14:00)接虫,烟粉虱在多毛番茄LA2329上的平均产卵量显著低于栽培番茄9706上的产卵量(分别为11粒,164粒).羽化后,烟粉虱雌虫在多毛番茄LA1777上的寿命显著低于在栽培番茄Moneymaker上的存活寿命(分别为5d,22d);而羽化后雌虫在LA1777上的平均产卵量显著低于在栽培番茄早粉2号上的产卵量(分别为7粒/头,95粒/头).在其他参数,如体型大小、存活率、发育历期等,没有显著性的变化.结果显示,较多毛番茄而言,栽培番茄是烟粉虱的较好寄主.而且,在评价抗烟粉虱番茄材料时,平均产卵量、羽化后雌虫寿命及产卵量是3个有效的评价参数.     

Growth and Development of Romanomermis culicivorax In Vitro

Various combinations of vertebrate and invertebrate tissue culture and microbiological media were utilized in an attempt to culture Romanomermis culicivorax (Mermithidae: Nematoda) in vitro. Most media were unsuitable and caused nematodes to become lumpy, vacuolated, and granular. Slow and limited growth and development of internal structures of the nematodes were obtained with variously supplemented Grace''s tissue culture and Schneider''s Drosophila media. In an enriched Grace''s medium, development attained by the nematodes after 3-4 wk was comparable to 4-5-day-old parasites grown in vivo in the mosquito host, Culex pipiens. Two molts were observed in vitro. Maximum dimensions in vitro were 7.0-mm length and 87-μm width at the widest point. The stichosome, stichocytes, and trophosome developed prominently. A filiform tail and highly cuticularized tube persisted throughout the culture period in vitro.     

Hatch and Reproduction of Globodera tabacum tabacum in Response to Tobacco,Tomato, or Black Nightshade

The effects of broadleaf tobacco, tomato, and black nightshade on juvenile hatch and reproduction of Globodera tabacum tabacum were determined in laboratory and greenhouse experiments. Root exudates from nightshade stimulated greater egg hatch than those from either ''Rutgers'' tomato or ''86-4'' tobacco. Hatch was greater at higher proportions of root exudates for all three plant species. Root exudates from plants greater than 3 weeks old stimulated more hatch than younger plants. No regression relationships existed between plant age and nematode batch. In other experiments, hatch from eggs in cysts was higher for tomato and nightshade after 10 weeks in greenhouse pots compared to tobacco and bare soil. Numbers of second-stage juveniles in eggs in cysts produced from a previous generation on the same host were highest on nightshade and less on tomato and tobacco. Cysts of variable age recovered from field soil had increased hatch in both root exudates or water compared to recently produced cysts from plants in growth chambers. Globodera t. tabacum may be subject to both host and environmentally mediated diapause.     

Liquid Culture of the Entomogenous Nematode Steinernema feltiae with Its Bacterial Symbiont

The insect-parasitic nematode, Steinernema feltiae Filipjev strain 42, was reared in liquid culture along with its bacterial symbiont, Xenorhabdus nematophilus Thomas &Poinar. First-stage juveniles developed into reproducing adults in a maintenance salts medium containing resuspended Xenorhabdus cells and the yeast Kluyveromyces marxianus (Hansen) van der Walt or cholesterol. Cultures with media depths greater than 4 mm required aeration. Nematode populations increased as bacterial density increased. An optimal culture system was obtained when the bacteria and nematodes developed in a semidefined medium containing tryptic soy, yeast extract, and cholesterol and were incubated on a rotary shaker at 25 ± 1 C. Under these conditions, up to 86% of the final population were infective juveniles.     

Histopathology of Okra and Ridgeseed Spurge Infected with Meloidodera charis

Histological observations of okra Abelomoschus esculentus ''Clemson Spineless'' and ridgeseed spurge Euphorbia glyptosperma (a common weed) infected with Meloidodera charis Hopper, indicated that the juvenile nematode penetrated the roots intercellularly. Within 5 days after plant emergence the nematode positioned its body in the cortical tissue parallel to the vascular system. By 10 days after plant emergence the juvenile had extended its head into the vascular system and initiated giant cell formation, generally in protophloem tissue. Giant cells were one celled and usually multi-nucleate. Eggs were observed in the female body 30 days after plants emerged and juveniles were found within the female body by 40 days. Nematode development progressed equally in the root system of either host plant. Generally, throughout the nematode''s life cycle its entire body remained inside the cortical tissue of okra. In ridgeseed spurge, however, the posterior portion of the female erupted through the host epidermis as early as 15 days after plant emergence; only the head and neck remained embedded in the host. The nematode caused extensive tissue disruption in the cortical and vascular system of both plant species. Corn, Zea mays, was another host of the nematode.     

Host Status of 'SeaIsle 1' Seashore Paspalum (Paspalum vaginatum) to Belonolaimus longicaudatus and Hoplolaimus galeatus

Belonolaimus longicaudatus and Hoplolaimus galeatus are considered among the most damaging pathogens of turfgrasses in Florida. However, the host status of seashore paspalum (Paspalum vaginatum) is unknown. Glasshouse experiments were performed in 2002 and 2003 to determine the tolerance of ''SeaIsle 1'' seashore paspalum to a population of B. longicaudatus and a population of H. galeatus, and to compare to ''Tifdwarf'' bermudagrass for differences. Both nematode species reproduced well on either grass, but only B. longicaudatus consistently reduced root growth as measured by root length. Belonolaimus longicaudatus reduced root growth (P ≤ 0.05) by 35% to 45% at 120 days after inoculation on both grasses. In 2003, higher inoculum levels of H. galeatus reduced root growth (P ≤ 0.05) by 19.4% in seashore paspalum and by 14% in bermudagrass after 60 and 120 days of exposure, respectively. Percentage reductions in root length caused by H. galeatus and B. longicaudatus indicated no differences between grass species, although Tifdwarf bermudagrass supported higher soil population densities of both nematodes than SeaIsle 1 seashore paspalum.     

Meloidogyne javanica Parasitic on Peanut

Peanut fields in four governorates of Egypt were surveyed to identify species of Meloidogyne present. Fourteen populations obtained from peanut roots were all identified as M. javanica based on perineal patterns, stylet and body lengths of second-stage juveniles, esterase phenotypes, and restriction fragment length polymorphisms of mtDNA. Three of 14 populations, all from contiguous fields in the Behara governorate, had individuals with a unique two-isozyme esterase phenotype. All populations of M. javanica tested on peanut had levels of reproduction on the M. arenaria-susceptible peanut cultivar Florunner that were not different from M. arenaria (P = 0.05), and had lower levels of reproduction on the M. arenaria-resistant genotype TxAG-7 than on Florunner (P = 0.05). Reproduction of the five Egyptian populations of M. javanica tested was lower on root-knot nematode resistant tomato cultivars Better Boy and Celebrity than on the root-knot nematode susceptible cultivar Rutgers (P = 0.05). These data are evidence that some populations of M. javanica are parasitic on peanut and that the peanut and tomato genotypes resistant to M. arenaria are also resistant to these populations of M. javanica.     

Greenhouse Studies on the Effect of Marigolds (Tagetes spp.) on Four Meloidogyne Species

The effects of preplanted marigold on tomato root galling and multiplication of Meloidogyne incognita, M. javanica, M. arenaria, and M. hapla were studied. Marigold cultivars of Tagetes patula, T. erecta, T. signata, and a Tagetes hybrid all reduced galling and numbers of second-stage juveniles in subsequent tomato compared to the tomato-tomato control. All four Meloidogyne spp. reproduced on T. signata ''Tangerine Gem''. Several cultivars of T. patula and T. erecta suppressed galling and reproduction of Meloidogyne spp. on tomato to levels lower than or comparable to a fallow control. Phytotoxic effects of marigold on tomato were not observed. Several of the tested marigold cultivars are ready for full-scale field evaluation against Meloidogyne spp.     

Host Differences among Florida Populations of Belonolaimus longicaudatus Rau

Three populations of Belonolaimus longicaudatus from Gainesville, Fuller''s Crossing, and Sanford, all in Florida, were tested for parasitism on Citrus jambhiri (rough lemon), Arachis hypogaea (''Early Runner'' peanut), Fragaria sp. (''Florida 90'' hybrid strawberry), and Lycopersicon esculentum (''Rutgers'' tomato). The three populations were found to be three different physiological races because: (i) the Fuller''s Crossing population reproduced well on and caused injury to rough lemon and tomato but not strawberry; (ii) the Gainesville population reproduced well on and caused injury to peanut, strawberry and tomato but not on rough lemon; (iii) the Sanford population reproduced well on and caused injury to peanut, reproduced well on tomato, but neither reproduced upon nor caused injury to strawberry or rough lemon. Morphologically, females of the Sanford population have a lower "c" value and a greater number of tail annules than those of the Gainesville and Fuller''s Crossing populations. Females and males of the Fuller''s Crossing population had higher "a" values than those of the other two populations.