Interaction of Pratylenchus penetrans and Rhizoctonia fragariae in Strawberry Black Root Rot

A split-root technique was used to examine the interaction between Pratylenchus penetrans and the cortical root-rotting pathogen Rhizoctonia fragariae in strawberry black root rot. Plants inoculated with both pathogens on the same half of a split-root crown had greater levels of root rot than plants inoculated separately or with either pathogen alone. Isolation of R. fragariae from field-grown roots differed with root type and time of sampling. Fungal infection of structural roots was low until fruiting, whereas perennial root colonization was high. Isolation of R. fragariae from feeder roots was variable, but was greater from feeder roots on perennial than from structural roots. Isolation of the fungus was greater from structural roots with nematode lesions than from non-symptomatic roots. Rhizoctonia fragariae was a common resident on the sloughed cortex of healthy perennial roots. From this source, the fungus may infect additional roots. The direct effects of lesion nematode feeding and movement are cortical cell damage and death. Indirect effects include discoloration of the endodermis and early polyderm formation. Perhaps weakened or dying cells caused directly or indirectly by P. penetrans are more susceptible to R. fragariae, leading to increased disease.     

Age Structure and Community Diversity of Nematodes Associated with Maize in Iowa Sandy Soils

Age structure of nematode populations around maize growing in sandy soils in Iowa was studied at soil depths of 0-15and 15-30 cm for 2 years. Numbers of Longidorus breviannulatus were generally greater at 0-15 cm than at 15-30 cm deep until mid to late season. The decline in numbers of females as the season progressed indicates that fecundity slowed and is evidence of only one generation per year. Peak populations of Pratylenchus scribneri and Xiphinema americanum occurred in late August or early September. Adults of Hoplolaimus galeatus were few in the roots but common in the soil, indicating that fertilization occurred mostly in the soil. Numbers of P. scribneri were generally greater at the lower depth, especially late in the season. Community diversity (H'') was less when nematode biomass was used instead of numbers. Numbers of H. galeatus did not decline over the winter. Numbers of L. breviannulatus, P. scribneri, and X. americanum declined significantly over the winter, but not between spring cultivation and planting.     

Artificial Feeding Systems for Plant-Parasitic Nematodes

Feeding of an "obligate" plant-parasitic nematode (nonfungal feeder), Pratylenchus scribneri, in the absence of plant tissue was demonstrated in an artificial system consisting of liquid media and indicator dyes including amaranth and various nontoxic food colors. Among the compounds tested, sucrose, dextrose, Gamborg''s B5 medium, and DL-methionine stimulated a small percentage of feeding (12-36%). A high percentage of feeding (90-100%) occurred in a filtrate from excised corn roots cultured in Gamborg''s B5 medium. This feeding system has the potential to develop an artificial medium for plant-parasitic nematodes and to screen novel nematicides that are stomach poisons.     

Pathogenicity and Histopathology of Rotylenchulus reniformis Infecting Cantaloup

Rotylenchulus reniformis was pathogenic to cantaloup (Cucumis melo ''Perlita'') under greenhouse conditions. These findings confirm field symptoms of cantaloup infected with R. reniformis. Histopathological studies show that the nematode penetrates the cortex perpendicular to the vascular system and comes to rest with the head against the endodermis in young roots. Feeding stimulated the pericycle to either side of the endodermal feeding cell and caused cell hypertrophy with enlargement of the nucleoli and granular thickening of the cytoplasm. In older roots where the endodermis had collapsed, the nematode fed directly into the pericycle and caused similar symptoms. Nematode development was more rapid at 27 C than at 21 C.     

The Effects of Temperature on Pratylenchus scribneri and P. alleni Populations on Soybeans and Tomatoes

In soil temperature tests, rates of Pratylenchus scribneri and P. alleni reproduction were measured at various lemperatures on ''Clark 63'' and ''Cutler 71'' soybeans and ''Rutgers'' tomatoes. Recovered P. scribneri equaled or exceeded initial inoculum levels at temperatures of 27.5 C or higher on soybeans, and at 20 C or higher on tomatoes. Population increases were greatest at 3.5 C on both hosts. Populations increased on soybeans, but not on tomatoes, when soil temperature was raised from 25 to 35 C for either 3 or 9 days. Recovered P. alleni were less than the initial inoculum at 27.5 C but higher at 32 and 37.5 C and at 27.5 C on tomatoes, the lowest temperature tested for this nematode. In the field, soil temperatures 10 cm deep in eastern Kansas soybean growing areas reach 27.5 C only occasionally and for relatively short periods, which probably explains the relatively low and variable populations of P. scribneri and P. alleni on soybeans there.     

Interactions Among Pratylenchus penetrans,P. scribneri,and Verticillium dahliae in the Potato Early Dying Disease Complex

Microplots were infested with combinations of the fungus Verticillium dahliae and Pratylenchus penetrans and P. scribneri to test for individual and combined effects of these organisms on potato yield and nematode reproduction. Verticillium dahliae alone caused yield losses in all 3 years of the experiment, and the interaction between P. penetrans and V. dahliae was significant (P ≤ 0.05) in 2 years. Pratylenchus penetrans alone caused yield losses in 2 years and P. scribneri alone caused yield losses in 1 year. No two-way or three-way interaction was found involving P. scribneri. In 1987, reproduction for low densities of P. penetrans was 5 times higher when P. scribneri was also present than when it was absent, and 3.5 times higher in 1988. In nematode species mixtures, reproduction of P. scribneri was decreased by V. dahliae in 1987-88. The final population density of P. scribneri was negatively affected by V. dahliae and positively related to the initial proportion of P. scribneri to P. penetrans. In species mixtures with proportions of P. penetrans ranging from 0.1 to 0.5, reproduction of P. penetrans was negatively affected by V. dahliae and decreased linearly in relation to the increase in the initial proportion of P. penetrans in both years. The final population density of P. penetrans was affected only by V. dahliae.     

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.     

Diagnostic Probes Targeting the Major Sperm Protein Gene That May Be Useful in the Molecular Identification of Nematodes

Discrimination of closely related nematode species is typically problematic when traditional identification characteristics are prone to intraspecific variation. In this study, a molecular approach that can distinguish Pratylenchus penetrans and P. scribneri is described. The approach uses universal primers in conjunction with polymerase chain reaction (PCR) to amplify equivalent fragments of the major sperm protein (msp) gene from any nematode. This gene fragment typically includes an intron of variable sequence. The presence of this highly variable segment in an otherwise conserved gene sequence allows P. penetrans and P. scribneri to be distinguished by either a species-specific amplification or by dot-blot hybridization. The approach is potentially of general utility in species-specific identification of nematodes.     

Some Ultrastructural Changes Induced in Resistant and Susceptible Soybean Roots Following Infection by Rotylenchulus reniformis

A developmental electron microscopic study of the parasitism of Rolylenchulus reniforrnis in resistant ''Peking'' and susceptible ''Lee'' soybeans was made during a 21-day period under controlled conditions. Within 2 days of inoculation, the nematode had penetrated the cortical cells to the endodermis where it inserted its stylet, secreted and initiated syncytial formation and cell hypertrophy. Syncytia primarily involved pericycle tissues and, to a lesser extent, xylem parenchyma and endodermis. When identifiable, the cell into which the nematode stylet was inserted to initiate syncytial development was endodermal. Susceptible tissues exhibited two basic phases of development during this infection period: (i) an initial phase represented by partial cell wail lysis and separation; and (ii) an anabolic phase, characterized by organelle proliferation and development accompanied by secondary wall deposits, which provided nutrition for sessile female development. The resistant or hypersensitive reaction (HR) lacked the anabolic phase found in the susceptible reaction, and was characterized by an extension and usually accelerated type of Iysis found in the first phase of the syncytial development. The HR was usually very evident 4 days after inoculation, and could be identified by an almost complete lysis of the cell walls and cytoplasm. The possibility that the initial cell of the developing syncytium or "prosyncyte" may influence a susceptible or resistant reaction is discussed. Successive stages of cell wall dissolution and the deposition of secondary cell walls are described.     

Influence of Temperature on Population Development of Eight Species of Pratylenchus on Soybean

In a soil temperature study, population increase on ''Clark 63'' soybeatt was most rapid at 30 C in Pratylenchus alleni, P. brachyurus, P. cofleae, P. neglectus, P. scribneri, and P. zeae and at 25 C in P. penetrans and P. vulnus. The last two were the only species that reproduced at 15 C. Populations of all species increased over the range of 20-30 C, except those of P. neglectus at 20 C and P. coffeae, which was not tested below 25 C. Only P. brachyurus, P. neglectus, P. scribneri and P. zeae reproduced at 35 C. At their optimum temperatures, P. scribneri exhibited the greatest population increase, 1248-fold, and P. penetrans the least, 32-fold. This is the first report of soybean as a host for P. vulnus.     

Field Interrelationships among Heterodera glycines,Pratylenchus scribneri,and Three Other Nematode Species Associated with Soybean

Field experiments were conducted in 1982 and 1983 to assess interactions between Heterodera glycines and Pratylenchus scribneri on soybean in southern Illinois. Soybean cyst nematode susceptible cultivar Williams 79 and resistant cultivar Fayette were treated or not treated with aldicarb 15G. Initial population densities were 35 H. glycines cysts containing eggs, 100 P. scribneri, 30 Helicotylenchus pseudorobustus, 225 Paratylenchus projectus, and 85 Tylenchorhynchus martini per 250 cm³ soil in 1982, whereas in 1983 populations were 11 H. glycines cysts, 330 P. scribneri, and 620 H. pseudorobustus. In both years H. glycines populations increased on nontreated Williams 79, decreased on both treated and nontreated Fayette, and remained at initial levels on treated Williams 79. Recovery of P. scribneri per gram dry root was different between nontreated cultivars in 1982 but not in 1983. Aldicarb treatment suppressed soil and root populations of P. scribneri on both cultivars in both years. Populations of H. pseudorobustus, P. projectus, and T. martini at harvest indicated little population increase on either nontreated cultivar. In 1982 H. glycines caused yield suppression but P. scribneri did not, as differences in yield occurred between cultivars but not between aldicarb treatments. In 1983, however, there were no yield differences between cultivars, but aldicarb application resulted in yield increase in both cultivars. In 1983 the yield increase resulting from P. scribneri control was approximately 25%. No synergistic effect on yield was observed between H. glycines and P. scribneri.     

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.     

Casparian bands occur in the periderm of Pelargonium hortorum stem and root

Background and Aims

Casparian bands are characteristic of the endodermis and exodermis of roots, but also occur infrequently in other plant organs, for example stems and leaves. To date, these structures have not been detected in phellem cells of a periderm. The aim of this study was to determine whether Casparian bands occur in phellem cells using tests that are known to detect Casparian bands in cells that also contain suberin lamellae. Both natural periderm and wound-induced structures were examined in shoots and roots.

Methods

Using Pelargonium hortorum as a candidate species, the following tests were conducted: (1) staining with berberine and counterstaining with aniline blue, (2) mounting sections in concentrated sulphuric acid and (3) investigating the permeability of the walls with berberine as an apoplastic, fluorescent tracer.

Key Results

(1) Berberine–aniline blue staining revealed a modification in the radial and transverse walls of mature phellem cells in both stems and roots. Three days after wounding through to the cortex of stems, the boundary zone cells (pre-existing, living cells nearest the wound) had developed vividly stained primary walls. By 17 d, staining of mature phellem cells of wound-induced periderm was similar to that of natural periderm. (2) Mature native phellem cells of stems resisted acid digestion. (3) Berberine was excluded from the anticlinal (radial and transverse) walls of mature phellem cells in stems and roots, and from the wound-induced boundary zone.

Conclusions

Casparian bands are present in mature phellem cells in both stems and roots of P. hortorum. It is proposed that Casparian bands act to retard water loss and pathogen entry through the primary cell walls of the phellem cells, thus contributing to the main functions of the periderm.     

Distribution of Pratylenchus scribneri between Root and Soil Habitats

The abundance of Pratylenchus scribneri in soil and root habitats was compared in potato and corn plots during 1986-88. Nematodes were extracted from 100-cm³ soil samples and the roots contained within the samples. The percentage of the population recovered from soil, similar among years and crops, averaged ca. 50% at the beginning and end of the growing season and ca. 20% from early to late season. Proportionately more adults and fourth-stage juveniles than younger stages were located outside roots until harvest. In a related study, nematodes were isolated from the roots, root surfaces, and soil associated with roots of whole corn and potato plants sampled from the field. Nematode population estimates calculated from the whole plant samples were generally lower than those based on soil cores, but showed similar patterns of population growth. Nematode density per gram dry weight was highest in roots, intermediate on root surfaces, and lowest in soil. Estimates of the absolute abundance of nematodes in each of the three habitats were highest in roots or soil, depending on the sampling date, and lowest on root surfaces. This study demonstrates that P. scribneri inhabits soil environments even when host roots are present and illustrates the importance of considering all possible habitats when estimating the size of Pratylenchus spp. populations.     

Pathogenicity and Interaction of Three Nematode Species on Six Bermudagrasses

The ring nematode (Criconemoides ornatus), stunt nematode (Tylenchorhynchus martini), and sting nematode (Belonolaimus Iongicaudatus) reproduced readily on six bermudagrasses (Common, ''U-3'', ''Tufcote'', ''Continental'', ''Tiffine'', and ''Tifdwarf''). Populations of a single nematode species influenced the population development of a second and third parasitic nematode species on a particular host plant. Activity of most nematodes adversely affected reproduction of other nematode species in mixed cultures. Generally, the number of fibrous roots produced by plants decreased as the number of nematode species in the treatments increased. Tifdwarf bermudagrass appeared to be more tolerant to C. ornatus and T. martini than other grasses tested.     

Anatomical Response of Grain Sorghum Roots to Meloidogyne incognita acrita

The cotton root-knot nematode, Meloidogyne incognita acrita, reproduced on the roots of grain sorghum, causing syncytia in the cortex or stele of lateral roots. Giant cells developed either singly with few nuclei or in groups with many nuclei. Giant cells that developed in groups appeared the same as those which developed singly. The pericycle and endodermis were interrupted at the site of nematode invasion. Large areas of these tissues were absent for one-third of the circumference of the stele and extended 1.5 mm longitudinally along the root. In the area where pericycle and endodernris were absent, the parenchyma of the cortex extended to the vascular elements, and abnormal xylem surrounding giant cells extended into the region of the cortex. Root-knot galls appeared on sorghum roots as elongate swellings, discrete knots, or swellings with root proliferation. Galls were not observed on brace roots.     

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.     

Comparison of Compatible and Incompatible Response of Potato to Meloidogyne incognita

One susceptible (D6) and two resistant (E2 and N4) clones of Solanum sparsipilum × (S. phureja × haploid of S. tuberosum) were used to study the responses of potato roots and tubers to race 1 of Meloidogyne incognita (Kofoid &White) Chitwood. The compatible response was characterized by rapid penetration of large numbers of second-stage juveniles (J2) into roots, cessation of root growth, and occasional curving of root tips. The life cycle of M. incognita in the susceptible clone was completed in 25 days at 23-28 C. The incompatible response was characterized by penetration of fewer J2 into roots, necrosis of feeding sites within 2-7 days, and lack of nematode development. There were no differences in response of tubers from resistant and susceptible clones to nematode infection. Small numbers of J2 were detected in tubers, but they did not develop.     

Feeding of the Nematode Acrobeloides nanus on Bacteria

Information on the effect of bacteria-feeding nematodes on bacterial populations in the soil is sparse. We have isolated, cultured, and microscopically examined bacteria and nematodes coexisting within an agricultural soil and have studied their feeding relationship. The bacterium Pseudomonas corrugata isolate 2140R is a biocontrol agent against the pathogenic fungus Gaeumannomyces graminis var. tritici. The nematode Acrobeloides nanus is a cosmopolitan, bacteria-feeding organism widespread in agricultural and arid soils throughout Australia. Using light and electron microscopy, we observed the ingestion and breakdown of P. corrugata in the pharynx of A. nanus and bacterial passage through the nematode intestine as well as the accumulation of fluorescent compounds from ingested and broken P. fluorescens in the lumen of the nematode''s intestine. We also observed A. nanus feeding, growing, and reproducing on the Gram-positive bacterium Clavibacter toxicus, the causative agent of the disease annual ryegrass toxicity, and detected crushed bacteria in the nematode''s intestine.     

Histological Observations of Rotylenchulus reniformis on Gossypium longicalyx and Interspecific Cotton Hybrids

Observations on the development of reniform nematode (Rotylenchulus reniformis) on roots of Gossypium longicalyx, G. hirsutum, and two interspecific hybrids derived from them were made by light microscopy. Gossypium longicalyx is reported to be immune to reniform nematode, but the mechanism(s) for resistance are unknown. Penetration of G. longicalyx roots by female nematodes was confirmed, and incipient swelling of the females, indicating initiation of maturation of the reproductive system, was observed. Female maturation occurred up to the formation of a single embryo inside the female body but not beyond this point. In both hybrids, development was inhibited but progressed further than in the immune parent. Reactions ranged from highly compatible, with the formation of active syncytia and full development of females, to incompatible with little or no development of the female. Compatible plants showed characteristic hypertrophied cells, enlarged nuclei, dense cytoplasm, and partial dissolution of cell walls, whereas incompatible plant reactions included lignification of the cells adjacent to the nematode head, or the complete collapse and necrosis of the cells involved. The need to characterize reactions and to carefully select among the plants descended from the hybrids during the introgression process, as well as the importance of combining the results of reproduction tests with histological observation of the plant-nematode interactions, is discussed.