Cellular Responses of Resistant and Susceptible Soybean Genotypes Infected with Meloidogyne arenaria Races 1 and 2

The cellular responses induced by Meloidogyne arenaria races 1 and 2 in three soybean genotypes, susceptible CNS, resistant Jackson, and resistant PI 200538, were examined by light microscopy 20 days after inoculation. Differences in giant-cell development were greater between races than among the soybean genotypes. M. arenaria race 1 stimulated small, poorly formed giant-cells in contrast with M. arenaria race 2, which induced well-developed, thick-walled, multinucleate giant-cells. The number of nuclei per giant-celt was variable, but fewer nuclei were usually present in giant-cells induced by race 1 (mean 16 nuclei) than in giant-cells induced by race 2 (mean 41 nuclei). Differences observed in giant-cell development were related to differences in growth and maturation of M. arenaria races 1 and 2 and host suitability of the soybean genotypes.     

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.     

Penetration Rates by Second-stage Juveniles of Meloidogyne spp. and Heterodera glycines into Soybean Roots

The rates of soybean root penetration by freshly hatched second-stage juveniles (J2) of Meloidogyne arenaria, M. hapla, M. incognita, M. javanica, and Heterodera glycines races 1 and 5 were examined over a period of 1 to 240 hours. Heterodera glycines entered roots more quickly than Meloidogyne spp. Penetration by most nematodes was accomplished within 48 hours. The increases in penetration after 48 hours were insufficient to warrant further assessments. Penetration of J2 into roots of soybean seedfings in a styrofoam container was as good or better than in a clay pot. Thus, rapid and accurate root-penetration assessments can be made at 48 hours after inoculation.     

Variability among Populations of Meloidogyne arenaria

Variability in reproduction and pathogenicity of 12 populations of Meloidogyne arenaria race 1 was evaluated on Florunner peanut, Centennial soybean, Rutgers tomato, G70, K326, and Mc944 tobacco, and Carolina Cayenne, Mississippi Nemaheart, and Santanka pepper. Differences among M. arenaria populations in rates of egg production 45 days after inoculation were observed for all cultivars except Santanka pepper. Differences among populations in dry top weights or fresh root weights were recorded on all cultivars. Numbers of nematode eggs produced on Florunner peanut varied from 3,419 to 11,593/g fresh root weight. On resistant tobacco cultivars (G70 and K326), one nematode population produced high numbers of eggs (12,042 and 6,499/g fresh root weight on G70 and K326, respectively), whereas the other populations produced low numbers of eggs (less than 500 eggs/g fresh root weight on both cultivars). Two variant M. arenaria race 1 populations were identified by factor analysis of reproductive rates on all nine cultivars. Differences m reproduction and pathogenicity observed among populations would affect the design of sustainable management systems for M. arenaria.     

Host-Parasite Relationships of Meloidogyne arenaria and M. incognita on Susceptible Soybean

Pathogenicity and reproduction of single and combined populations of Meloidogyne arenaria and M. incognita on a susceptible soybean (Glycine max cv. Davis) were investigated. Significant galling and egg mass production were observed on roots of greenhouse-grown soybean inoculated with M. arenaria and M. incognita, in combination and individually. M. arenaria produced more galls and egg masses than M. incognita, whereas in combined inoculation with both nematode species, gall and egg production was intermediate. In growth chamber tests, inoculations with M. arenaria and M. incognita, singly or in combination, produced more galls and egg masses at 30 C than at 25 C. At 25 C, M. arenaria alone produced significantly more galls and egg masses than the combined M. arenaria plus M. incognita, while M. incognita produced the fewest. At 30 C, numbers of egg masses produced by M. arenaria did not differ significantly from combined M. arenaria and M. incognita. In temperature tank tests, M. incognita produced more galls and egg masses at 28 C than at 24 C soil temperature. In contrast, numbers of galls, egg masses, and eggs of M. arenaria were slightly higher at 24 C than at 28 C. Combined inoculum of both nematode species produced greater numbers of galls at 24 C than at 28 C.     

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.     

Interaction of Vesicular-Arbuscular Mycorrhizae and Cultivars of Alfalfa Susceptible and Resistant to Meloidogyne hapla

The interaction between vesicular-arbuscular mycorrhizal (VAM) fungi and the root-knot nematode (Meloidogyne hapla) was investigated using both nematode-susceptible (Grasslands Wairau) and nematode-resistant (Nevada Synthetic XX) cultivars of alfalfa (Medicago sativa) at four levels of applied phosphate. Mycorrhizal inoculation improved plant growth and reduced nematode numbers and adult development in roots in dually infected cultures of the susceptible cultivar. The tolerance of plants to nematode infection and development when preinfected with mycorrhizal fungi was no greater than when they were inoculated with nematodes and mycorrhizal fungi simultaneously. Growth of plants of the resistant cultivar was unaffected by nematode inoculation but was improved by mycorrhizal inoculation. Numbers of nematode juveniles were lower in the roots of the resistant than of the susceptible cultivar and were further reduced by mycorrhizal inoculation, although no adult nematodes developed in any resistant cultivar treatment. Inoculation of alfalfa with VAM fungi increased the tolerance and resistance of a cultivar susceptible to M. hapla and improved the resistance of a resistant cultivar.     

Mechanism of Resistance to Meloidogyne arenaria in the Peanut Cultivar COAN

Resistance to Meloidogyne arenaria in the peanut cultivar COAN is inherited as a single, dominant gene. The mechanism of resistance to M. arenaria in COAN was evaluated in three experiments. In the first experiment the number of second-stage juveniles (J2) of M. arenaria penetrating roots of the susceptible cultivar Florunner was higher than the number of J2 penetrating roots of the resistant peanut cultivar COAN (P < 0.05). In a second experiment it was determined that the root size and number of potential infection courts (root tips) were similar for the two peanut cultivars. The number of nematodes emigrating from roots of COAN after penetration was greater than emigrated from roots of Florunner (P < 0.05). Necrotic host tissue was rarely observed in roots of COAN infected with M. arenaria, suggesting that resistance to M. arenaria does not involve a necrotic, hypersensitive response. Most of the J2 observed in roots of COAN were restricted to the cortical tissue, with only 1 of 90 J2 observed being associated with the vascular cylinder, whereas in Florunner >70% of the J2 were associated with vascular tissues. Resistance in COAN may be due to constitutive factors in the roots.     

Aggressiveness and Reproduction of Four Meloidogyne arenaria Populations on Soybean

Aggressiveness and reproduction differed among four geographical populations of M. arenaria on six soybean cultivars in field microplots. These differences were consistent over 3 years. The populations did not differ in virulence; i.e., population by cultivar interactions were not significant. Perineal pattern morphology, the North Carolina differential host test, chromosome counts of immature oocytes, and esterase phenotypes confirmed that the four populations were M. arenaria. Three populations were host race 2 and one population was host race 1.     

Effects of Tropical Rotation Crops on Meloidogyne arenaria Population Densities and Vegetable Yields in Microplots

The effects of 12 summer crop rotation treatments on population densities of Meloidogyne arenaria race 1 and on yields of subsequent spring vegetable crops were determined in microplots. The crop sequence was: (i) rotation crops during summer 1991 ; (ii) cover crop of rye (Secale cereale) during winter 1991-92; (iii) squash (Cucurbita pepo) during spring 1992; (iv) rotation crops during summer 1992; (v) rye during winter 1992-93; (vi) eggplant (Solanum melongena) during spring 1993. The 12 rotation treatments were castor (Ricinus communis), cotton (Gossypium hirsutum), velvetbean (Mucuna deeringiana), crotalaria (Crotalaria spectabilis), fallow, hairy indigo (Indigofera hirsuta), American jointvetch (Aeschynomene americana), sorghum-sudangrass (Sorghum bicolor x S. sudanense), soybean (Glycine max), horsebean (Canavalia ensiformis), sesame (Sesamum indicum), and peanut (Arachis hypogaea). Compared to peanut, the first eight rotation treatments resulted in lower (P ≤ 0.05) numbers of M. arenaria juveniles on most sampling dates. Soybean, horsebean, and sesame rotations were less effective in suppressing nematodes. Yield of squash was greater (P ≤ 0.05) following castor, cotton, velvetbean, and crotalaria than following peanut. Compared to the peanut rotation, yield of eggplant was enhanced (P ≤ 0.10) following castor, crotalaria, hairy indigo, American jointvetch, and sorghum-sudangrass. Several of these rotation crops may provide a means for depressing M. arenaria population densities on a short-term basis to enhance yields in a subsequent susceptible vegetable crop.     

Penetration and Post-infectional Development and Reproduction of Meloidogyne arenaria Races 1 and 2 on Susceptible and Resistant Soybean Genotypes

Penetration, post-infectional development, reproduction, and fecundity of Meloidogyne arenaria races 1 and 2 were studied on susceptible (CNS), partially resistant (Jackson), and highly resistant (PI 200538 and PI 230977) soybean genotypes in the greenhouse. The ability to locate and invade roots was similar between races, but more juveniles penetrated roots of susceptible CNS than the resistant genotypes. At 10 days after inoculation, 56% and 99% to 100% of race 1 second-stage juveniles were vermiform or sexually undifferentiated in CNS and the resistant genotypes, respectively. In contrast, only 2%, 42%, 44%, and 62% of race 2 juveniles had not initiated development in CNS, Jackson, PI 200538, and PI 230977, respectively. By 20 days after inoculation, 88% to 100% of race 2 nematodes in roots of all genotypes were females, whereas only 25% and 1% of race 1 were females in CNS and the resistant genotypes, respectively. For all four genotypes, race 1 produce 85% to 96% fewer eggs per root system 45 days after inoculation than race 2. At 45 days after inoculation race 2 produced more eggs on CNS than the other genotypes.     

Reproductive and Damage Potential of Ditylenchus destructor on Six Peanut Cultivars

Commercial peanut cultivars were evaluated for host suitability and sensitivity to Ditylenchus destructor. All cultivars were susceptible. Approximately 94% of the final population were in the pods. Highest Pf occurred at harvest on early maturing cultivars. Damage occurred on four of six cuttivars at Pi = 100/3 liters of soil and all six cultivars at Pi = 1,000. Norden and Selmani were the most susceptible cultivars. Sellie was the most tolerant and highest yielding cultivar. This cultivar may be the most profitable one for growers.     

Effects of Resistance in Phaseolus vulgaris on Development of Meloidogyne Species

Use of resistant Phaseolus vulgaris germplasm has a potential role in limiting damaging effects of Meloidogyne spp. on bean production. Effects of two genetic resistance systems in common bean germptasm on penetration and development of Meloidogyne spp. were studied under growth room conditions at 22°C to 25°C. Nemasnap (gene system 1) and G1805 (gene system 2) were inoculated with second-stage juveniles (J2) of M. incognita race 2 and M. arenaria race 1, respectively; Black Valentine was used as the susceptible control. Up to 7 days after inoculation, there were no differences in numbers of M. incognita J2 penetrating roots of Black Valentine and Nemasnap; subsequently, more nematodes were present in Black Valentine roots (P < 0.05). More nematodes reached advanced stages of development in Black Valentine than in Nemasnap roots (P < 0.05). Total numbers of M. arenaria were greater in Black Valentine than in G 1805 roots from 14 days after inoculation (P < 0.05). Advanced stages of development occurred earlier and in greater numbers in Black Valentine plants than in G1805 plants. In these studies, resistance to M. incognita race 2 and M. arenaria race 1 in bean germplasm, which contain gene system 1 and gene system 2, respectively, was expressed by delayed nematode development rather than by differential penetration compared with susceptible plants.     

Reproduction of Meloidogyne spp. on Resistant Peanut Genotypes from Three Breeding Programs

Three described species of root-knot nematode parasitize peanut (Arachis hypogaea): Meloidogyne arenaria race 1 (Ma), M. hapla (Mh), and M. javanica (Mj). Peanut cultivars with broad resistance to Meloidogyne spp. will be useful regardless of the species present in the field. The objective of this study was to determine whether peanut genotypes with resistance to M. arenaria originating from three different breeding programs were also resistant to M. hapla and M. javanica. The experiment used a factorial arrangement (completely randomized) with peanut genotype and nematode population as the factors. The five peanut genotypes were ''COAN'' and AT 0812 (highly resistant to Ma), C209-6-13 (moderately resistant to Ma), and ''Southern Runner'' and ''Georgia Green'' (susceptible to Ma). The four nematode populations were two isolates of Ma (Gibbs and Gop) and one isolate each of Mh and Mj. On COAN or AT 0812, both Ma and Mj produced <10% of the eggs produced on Georgia Green. On the peanut genotype C209-6-13, Ma and Mj produced about 50% of the eggs produced on Georgia Green. None of the resistant genotypes exhibited a high level of resistance to Mh. The lack of resistance to Mh in any cultivars or advanced germplasm is a concern because the identity of a Meloidogyne sp. in a particular peanut field is generally not known. Breeding efforts should focus on moving genes for resistance to M. hapla into advanced peanut germplasm, and combining genes for resistance to the major Meloidogyne spp. in a single cultivar.     

Damage Functions for Three Meloidogyne Species on Arachis hypogaea in Texas

The yield response of Florunner peanut to different initial population (Pi) densities of Meloidogyne arenaria, M. javanica, and an undescribed Meloidogyne species (isolate 93-13a) was determined in microplots in 1995 and 1996. Seven Pi''s (0, 0.5, 1, 5, 10, 50, and 100 eggs and J2/500 cm³ soil) were used for each Meloidogyne species in both years. The three species reproduced abundantly on Florunner in both years. In 1995, mean reproduction differed among the three species; mean Rf values were 10,253 for isolate 93-13, 4,256 for M. arenaria, and 513 for M. javanica. In 1996, the reproduction of M. arenaria (mean Rf = 7,820) and isolate 93-13a (mean Rf = 7,506) were similar, and both had greater reproduction on peanut than did M. javanica (mean Rf = 2,325). All three nematode species caused root and pod galling, and a positive relationship was observed between Pi and the percentage of pods galled. Meloidogyne arenaria caused a higher percentage of pod galling than did M. javanica or isolate 93-13a. A negative linear relationship between log₁₀ (Pi + 1) and pod yield was observed for all three nematode species each year. The yield response slopes were similar except for that of M. javanica, which was less negative than that of isolate 93-13a in 1995, and less negative than that of M. arenaria and isolate 93-13a in 1996.     

Identification of Sources of Resistance to Four Species of Root-knot Nematodes in Tobacco

Resistance to the southern root-knot nematode, Meloidogyne incognita races 1 and 3, has been identified, incorporated, and deployed into commercial cultivars of tobacco, Nicotiana tabacum. Cultivars with resistance to other economically important root-knot nematode species attacking tobacco, M. arenaria, M. hapla, M. javanica, and other host-specific races of M. incognita, are not available in the United States. Twenty-eight tobacco genotypes of diverse origin and two standard cultivars, NC 2326 (susceptible) and Speight G 28 (resistant to M. incognita races 1 and 3), were screened for resistance to eight root-knot nematode populations of North Carolina origin. Based on root gall indices at 8 to 12 weeks after inoculation, all genotypes except NC 2326 and Okinawa were resistant to M. arenaria race 1, and races 1 and 3 of M. incognita. Except for slight root galling, genotypes resistant to M. arenaria race 1 responded similarly to races 1 and 3 of M. incognita. All genotypes except NC 2326, Okinawa, and Speight G 28 showed resistance to M. javanica. Okinawa, while supporting lower reproduction of M. javanica than NC 2326, was rated as moderately susceptible. Tobacco breeding lines 81-R-617A, 81-RL- 2K, SA 1213, SA 1214, SA 1223, and SA 1224 were resistant to M. arenaria race 2, and thus may be used as sources of resistance to this pathogen. No resistance to M. hapla and only moderate resistance to races 2 and 4 of M. incognita were found in any of the tobacco genotypes. Under natural field infestations of M. arenaria race 2, nematode development on resistant tobacco breeding lines 81-RL-2K, SA 1214, and SA 1215 was similar to a susceptible cultivar with some nematicide treatments; however, quantity and quality of yield were inferior compared to K 326 plus nematicides.     

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.     

Partial Characterization of Cytosolic Superoxide Dismutase Activity in the Interaction of Meloidogyne incognita with Two Cultivars of Glycine max

The closely related soybean (Glycine max) cultivars Centennial and Pickett 71 were confirmed to be resistant and susceptible, respectively, to the root-knot nematode Meloidogryne incognita. Increases in superoxide dismutase (SOD) activity were detected in roots of both soybean cultivars 48 hours following inoculation. Superoxide dismutase activity increased in roots of the susceptible cultivar overall, but declined after 96 hours in roots of the resistant cultivar. The isoelectric points of SOD isolated from preparasitic and parasitic developmental stages of the nematode appeared to differ. The SOD activity increased dramatically as nematodes matured and enlarged. Plant and nematode SOD were present as ca. 40-kDa cuprozinc dimers. Initial increases in SOD activity in infected tissue appeared to involve nematode regulation of plant gene expression. However, as the nematode enlarged, SOD activity could be detected within the female body only.     

Persistence and Suppressiveness of Pasteuria penetrans to Meloidogyne arenaria Race

The long-term persistence and suppressiveness of Pasteuria penetrans against Meloidogyne arenaria race 1 were investigated in a formerly root-knot nematode suppressive site following 9 years of continuous cultivation of three treatments and 4 years of continuous peanut. The three treatments were two M. arenaria race 1 nonhost crops, bahiagrass (Paspalum notatum cv. Pensacola var. Tifton 9), rhizomal peanut (Arachis glabrata cv. Florigraze), and weed fallow. Two root-knot nematode susceptible weeds commonly observed in weed fallow plots were hairy indigo (Indigofera hirsuta) and alyce clover (Alysicarpus vaginalis). The percentage of J2 with endospores attached reached the highest level of 87% in 2000 in weed fallow, and 63% and 53% in 2002 in bahiagrass and rhizomal peanut, respectively. The percentage of endospore-filled females extracted from peanut roots grown in weed fallow plots increased from nondetectable in 1999 to 56% in 2002, whereas the percentages in bahiagrass and rhizomal peanut plots were 41% and 16%, respectively. Over 4 years, however, there was no strong evidence that endospores densities reached suppressive levels because peanut roots, pods, and pegs were heavily galled, and yields were suppressed. This might be attributed to the discovery of M. javanica infecting peanut in this field in early autumn 2001. A laboratory test confirmed that although the P. penetrans isolate specific to M. arenaria attached to M. javanica J2, no development occurred. In summary, P. penetrans increased on M. arenaria over a 4-year period, but apparently because of infection of M. javanica on peanut at the field site root-knot disease was not suppressed. This was confirmed by a suppressive soil test that showed a higher level of soil suppressiveness than occurred in the field (P ≤ 0.01).     

Impact of Herbicides on Heterodera glycines Susceptible and Resistant Soybean Cultivars

Several abiotic and biotic stresses can affect soybean in a growing season. Heterodera glycines, soybean cyst nematode, reduces yield of soybean more than any other pathogen in the United States. Field and greenhouse studies were conducted to determine whether preemergence and postemergence herbicides modified the reproduction of H. glycines, and to determine the effects of possible interactive stresses caused by herbicides and H. glycines on soybean growth and yield. Heterodera glycines reproduction factor (Rf) generally was less on resistant than susceptible cultivars, resulting in a yield advantage for resistant cultivars. The yield advantage of resistant cultivars was due to more pods per plant on resistant than susceptible cultivars. Pendimethalin reduced H. glycines Rf on the susceptible cultivars in 1998 at Champaign, Illinois, and in greenhouse studies reduced dry root weight of H. glycines-resistant and susceptible cultivars, therefore reducing Rf on the susceptible cultivars. The interactive stresses from acifluorfen or imazethapyr and H. glycines reduced the dry shoot weight of the resistant cultivar Jack in a greenhouse study. Herbicides did not affect resistant cultivars'' ability to suppress H. glycines Rf; therefore, growers planting resistant cultivars should make herbicide decisions based on weeds present and cultivar tolerance to the herbicide.