Evaluation of resistance to root-knot nematode (Meloidogyne incognita) in okra cultivars

The root-knot nematode, Meloidogyne incognita, is one of the major limiting factors affecting plant growth and yield causing an estimated $100 billion loss per year worldwide. Synthetic pesticides, though instantaneously effective, are usually prohibitively expensive, not readily available, may cause hazards to both man and livestock, and inflict injury to the environment. Notable among the alternatives to nematicides is the use of resistant cultivars which are inexpensive and eco-friendly. In the present studies, twelve okra (Abelmoschus esculentus L) cultivars were evaluated for their resistance against M. incognita under field conditions. Ten-day old okra plants of test cultivars were inoculated with 3000 freshly hatched second stage juveniles of M. incognita. The nematode caused reductions in various growth parameters of all the cultivars to varying levels over their respective controls. None of the cultivars was found completely resistant. The cultivar ‘Sharmeeli’ was highly susceptible as >100 galls were recorded on the roots. Sharmeeli also showed maximum reductions in growth among the cultivars evaluated. The cultivars Anmol and Sindha were susceptible with 71–100 galls. The cultivars Sabz Pari, Super Star, PMS-55 and PMS Beauty were moderately susceptible with 31–70 galls and comparatively less reductions in growth. Cultivars Sanam, Dikshah, Arka Anamika, Ikra-1 and Ikra-2 with 11–30 galls were rated as moderately resistant and showed less damage by the nematode as compared to susceptible cultivars and their planting could provide a useful tool to control root-knot nematodes.     

Response of selected cucumber cultivars to Meloidogyne incognita

Root-knot nematodes (Meloidogyne spp.) are one of the most destructive pathogens of vegetables. The cultivars resistant to root-knot nematodes have comparatively better crop yield than susceptible varieties and can be employed as a component of integrated nematode management. In the present study, 15 cultivars of cucumber (Cucumis sativus) were tested for their response to Meloidogyne incognita by inoculating with 3000 2nd stage juveniles of the nematode. All cultivars varied significantly in causing reductions in growth parameters over their controls. Likewise, all cultivars behaved differently regarding formation of galls and egg masses and reproductive factor. It was found that none of the tested cultivars was found immune or highly resistant. Long Green was the only cultivar which was found resistant against M. incognita as it showed minimum galls (8.2) and reductions in growth parameters. On the other hand, cultivars Mehran, Mirage, Thamin-II, and Royal Sluis were highly susceptible as evident by maximum galls (>100) on their roots and reductions in growth parameters. Similarly, the cultivars Green Wonder, Cucumber Citriolo and Poinsett were moderately susceptible (31–70 galls) while Babylon, Cobra and Falcon-560 (71–100 galls) were susceptible and reductions in growth parameters of these cultivars were less than those in highly susceptible cultivars. Four cultivars, Marketmore, Dynasty, Pioneer-II, and Summer Green, were rated as moderately resistant (11–30 galls) as these cultivars showed less damage by the nematode than moderately susceptible, susceptible and highly susceptible cultivars.     

Screening of three sweet potato (Ipomoea batatas L.) cultivars for resistance to different virulence groups of root-knot nematodes (Meloidogyne spp.) under controlled conditions

The susceptibility of three sweet potato cultivars (Ipomoea batatas L.) C4, TIS 3290 and TIS 9162 was evaluated against 156 isolates of Meloidogyne spp. with the aim to include resistant/tolerant sweet potato cultivars in a crop rotation scheme for the management of root-knot nematodes. The nematode isolates corresponded to races 1, 2 and 3 of Meloidogyne arenaria (n = 7), races 1, 2, 3 and 4 of M. incognita (n = 131) and Meloidogyne javanica (n = 18). Also, the isolates of M. incognita were differentiated in virulence groups: Pepper (n = 35), Pepper-Mi (n = 25), Tomato (n = 41) and Tomato-Mi (n = 30), depending on their ability to parasitize resistant pepper and tomato cultivars. The tested isolates of M. javanica parasitized C4 and TIS 3290, but not TIS 9162, whereas M. arenaria parasitized C4 and TIS 9162, but not TIS 3290, and M. incognita was able to parasitize the three sweet potato cultivars tested. C4 was the most susceptible cultivar to all nematode species tested, especially M. incognita, TIS 3290 was the most resistant and TIS 9162 was in between (7.2, 62.9 and 26.9% of resistant plants, respectively). Susceptibility of the sweet potato cultivars showed slight variations depending on the race or virulence group of M. incognita. The results suggest that sweet potato cultivars TIS 3290 and TIS 9162 may be used as rotation crops in fields where root-knot nematodes are present, their selection depending on the Meloidogyne isolates present. The use of resistant sweet potato cultivars would be preferably combined with other management practices to avoid virulence selection in nematode isolates.     

Infestation assessment of root-knot nematodes (Meloidogyne spp.) associated with cucumber in the Pothowar region of Pakistan

Studies to estimate root-knot nematode infestations on cucumber were conducted during mid- to late-season at 378 randomly selected sites in 126 villages of the four districts across the Pothowar zone of the country. The overall mean infestation of root-knot nematodes in the region was found to be 15.66%. The studies revealed variations in the incidence and severity of root-knot nematodes in the four districts. The incidence of root-knot nematodes was the highest in Rawalpindi district (21.86%), followed by 13.89% in Attock. Of the four districts, the minimum incidence of 10.97% was recorded in Jhelum district. The maximum mean severity (3.79), measured in terms of the galling index, was found in Rawalpindi district, while the minimum (1.86) was observed in Jhelum district. The mean severities of root-knot nematodes in the districts of Attock and Chakwal were 2.66 and 2.19 respectively. Variations in incidence and severity were also observed among subdivisions of the districts. Of all the associated species of root-knot nematodes, Meloidogyne incognita constituted 78.53%, Meloidogyne javanica 19.03%, Meloidogyne arenaria 1.82% and Meloidogyne hapla 0.62%. M. incognita and M. javanica were recorded in all of the districts, with M. incognita predominating. M. incognita as a pure population was recorded in 29.63% of the villages, while the other three species were found as mixtures. The most common mixed population was M. incognita and M. javanica, observed in 70.37% of villages in the region. M. arenaria and M. hapla were not found together in any of the population mixtures. The results indicate that cucumber is severely attacked by root-knot nematodes, warranting adoption of strict control measures for its management.     

Characterizing races of Meloidogyne incognita, M. javanica and M. arenaria in the West Mediterranean region of Turkey

A total of 95 samples of Meloidogyne incognita (60), Meloidogyne javanica (28) and Meloidogyne arenaria (7) collected from West Mediterranean region of Turkey were tested for identifying races of the nematodes using the North Carolina Differential Host Test. Races 2 and 6 of M. incognita were identified in 58 and 2 samples, respectively. Race 1 of M. javanica was identified from all 28 samples. Races 2 and 3 of M. arenaria were identified in 5 and 2 samples, respectively. The results also showed that two races of M. incognita and all races of M. javanica did not attack on pepper cv. California Wonder. These findings may contribute to improving resistant varieties and may also provide basis for developing a cropping system to control root-knot nematodes.     

Amendment of soil with African marigold and sunn hemp for management of Meloidogyne incognita in selected legumes

Field experiments were conducted in 2008 and 2009 in the tropical rainforest zone of Nigeria to investigate the effects of amendment of soil with seedlings of African marigold (Tagetes erecta) and sunn hemp (Crotalaria juncea) incorporated singly in plots on Meloidogyne incognita and yield of cowpea and soybean. The experimental field, which was naturally free of plant-parasitic nematodes, was inoculated with chopped roots of M. incognita race 2-infected Celosia argentea roots and planted to tomato to increase M. incognita population at the site.Eight week-old marigold seedlings were incorporated in cowpea or soybean field and eight week-old sunn hemp seedlings were also incorporated in cowpea or soybean field. At the ends of the experiments, M. incognita population densities were significantly higher in control plots than those of the plots amended with marigold or sunn hemp with correspondingly higher grain yield in the amended plots in both cowpea and soybean fields in both years. A significantly higher population of the nematode and consequently, lower yield was associated with cultivar Ife Brown than cultivar Ife Bimpe of cowpea for each treatment whereas in soybean cultivars, the pattern was not definite. Also twelve seedlings of marigold or sunn hemp per plot incorporated into the soil produced significantly higher grain yield in cultivar Ife Brown of cowpea and cultivar TGX 1440 of soybean compared to six seedlings per plot. The results of this study suggest that incorporating marigold or sunn hemp in M. incognita-infected cowpea or soybean field has potentials to suppress M. incognita population and reduce nematode damage on yield of the associated leguminous crops.     

Two-year investigations on the integrated control of weeds and root parasites in Virginia bright tobacco (Nicotiana tabacum L.) in central Italy

The current revision of pesticides in the EU, including those for soil disinfection, will necessitate the introduction of alternative integrated methods for the control of weeds and parasites in tobacco production in Italy. The present study compared the effect of some soil fumigants, biocontrol agents, cover crops and mulching for the control of tobacco weeds and root parasites. The results show that the different soil fumigants had an important role in increasing tobacco yield, due to their ability to control weeds and the nematodes Meloidogyne incognita and Meloidogyne javanica. In particular, a typical nematicide like 1,3-dichloropropene did not control only nematodes but also Portulaca oleracea, a common weed in Italian tobacco fields. Fenamiphos, although showed a certain efficacy in controlling root-knot nematodes, did not generally have a satisfactory activity at the dose of 40 g m⁻¹; cover crops like Trifolium squarrosum and Eruca sativa and a commercial formulation composed by the endomycorrhizal fungi Glomus spp., the antagonistic fungi Trichoderma spp. and bacteria including Pseudomonas, Bacillus and Streptomyces (Micosat F, CCS Aosta, Italy), were not able to control root tobacco diseases. The investigations confirmed that a prolonged monoculture causes a reduction of black root rot. Soil fumigation and the use of Good Agricultural Practices (GAP), influencing the inoculum density of Chalara populations, can contribute to the maintenance or to the establishment of particularly unfavourable conditions for the saprophytic growth of the pathogen, leading to a lower development of the disease.     

Cis- and trans-protopinium,a novel nematicide,for the eco-friendly management of root-knot nematodes

Plant-derived nematicides are being increasingly used while select synthetic nematicides are phased out for environmental concerns. This is the first report on the in vitro nematicidal activity of cis- and trans-protopinium isolated from roots of Fumaria parviflora and its in vivo effect on the southern root-knot nematode Meloidogyne incognita. Cis- and trans-protopinium was isolated from the methanolic fraction FM2.1, and its structure elucidated using ¹³C and ¹H nuclear magnetic resonance (NMR). The NMR spectra were characterized using deuterated dimethyl sulfoxide (DMSO) at temperatures of 25 and 80 °C. In an in vitro study, over 120 h of incubation, the area under cumulative percent hatch inhibition and mortality of cis- and trans-protopinium reached 100% at a concentration of 200 μg mL⁻¹. In the greenhouse and field settings, cis- and trans-protopinium was evaluated against M. incognita on tomato at a concentration of 100, 200, and 300 μg mL⁻¹ for two consecutive seasons, that is, spring and autumn, in 2010. At a concentration of 300 μg mL⁻¹ in the greenhouse and field trials during spring and autumn, cis- and trans-protopinium significantly reduced the nematode galling index, the number of females per gram of root, and the reproduction factor, as well as increased plant height, fresh and dry shoot weights, and root length. Therefore, cis- and trans-protopinium can be used as an effective and safe nematicide against M. incognita on tomato in an organic and sustainable agricultural production system. Phytochemicals have various agricultural applications, especially to control economically important nematode pests.     

Virulence response to the Mi.1 gene of Meloidogyne populations from tomato in greenhouses

In a comparison of reproductive traits of 29 field populations of Meloidogyne spp. on resistant and susceptible tomatoes, 48% of the populations were virulent against the Mi.1 gene. Virulent populations produced more (P < 0.05) egg masses/plant, eggs/plant, eggs/g root, and showed higher infection frequencies and multiplication rates than the avirulent populations. The fecundity of females did not differ between the resistant and susceptible genotype, and did not change with the virulence status of the populations. On the resistant genotype, the populations collected from the susceptible tomato showed lower (P < 0.05) reproductive traits than those collected from rootstocks followed by those from resistant tomato. Meloidogyne javanica produced more (P < 0.05) egg masses/plant and eggs/g root than Meloidogyne incognita on the resistant genotype. A time course experiment was conducted to determine if root penetration by M. javanica populations with different Mi.1 virulence status is conditioned by the expression of resistance on tomato. Significant differences between populations were observed as early as of 2 days post-inoculation on the susceptible genotype. On the resistant genotype, low juvenile numbers from populations with acquired virulence penetrated the roots up to 4 dpi, but their numbers significantly increased 7 dpi. Juveniles of the natural virulent population followed similar penetration pattern on the resistant and susceptible genotypes. Peroxidases are involved in the oxidative burst typically associated with the hypersensitive response in incompatible host–pathogen interactions. Isoelectrofocusing isozyme analysis revealed an increase in the peroxidase activity on the resistant genotype after inoculation with the avirulent population and to a lesser degree with the populations with acquired virulence in comparison with the natural virulent population.     

Managing Meloidogyne incognita and Bemisia tabaci with thiacloprid in cucumber crops in China

Field trials were conducted to determine the potential use of thiacloprid for integrated Meloidogyne incognita and Bemisia tabaci B biotype control in cucumber (Cucumis sativus Linn.) in China. The following five treatments were evaluated: three thiacloprid doses (7.5, 15, and 30 kg ha⁻¹), an avermectin treatment (7.5 L ha⁻¹) and an untreated control. All thiacloprid application rates were effective for reducing the M. incognita and B. tabaci B biotype populations. M. incognita was reduced by 51.0%∼86.7% on 60th day and B. tabaci B biotype was reduced by about 37.2%∼95.3% within 21 days, respectively. In addition, greater thiacloprid doses resulted in fewer nematodes and whiteflies. Over two successive years, the cucumber plants that were treated with thiacloprid at a dose of 15 kg ha⁻¹ had the greatest plant height, plant vigor and marketable yield, which were 134.1 cm, 91, 1514.3 g plant⁻¹, respectively in 2010–2011 and 151.9 cm, 93, 1651.4 g plant⁻¹, respectively in 2011–2012. In addition, thiacloprid was superior to avermectin. The results of this study demonstrated that thiacloprid could be used in cucumber production for the integrated control of M. incognita and B. tabaci B biotype. Furthermore, a dose of 15 kg ha⁻¹ of thiacloprid was recommended for controlling nematode and whitefly populations according to the control effect and marketable cucumber yield.     

Root-knot nematodes,a growing problem for Conilon coffee in Espírito Santo state,Brazil

Root-knot nematodes (Meloidogyne spp.) are non-quarantine pests that are subject to regulations in Brazil because they are limiting to coffee production and easily spread by planting infected seedlings. Containing their dissemination requires knowledge of their distribution in coffee-producing areas in order to establish phyto-sanitary measures. The object of this work is to evaluate the distribution of Meloidogyne spp. in Coffea spp. in Espírito Santo state, where coffee growing is expanding. Soil and root samples were collected in Coffea arabica and Coffea canephora Conilon. Identification of Meloidogyne spp. was carried out by esterase isoenzyme phenotype and by the host range. In Espírito Santo, Meloidogyneincognita, Meloidogyne exigua, and Meloidogyne paranaensis were identified. Phenotypes I1 and I2 of M. incognita were present in 21% of all the sampled properties. This species was found mainly in the Serrana region and north of the state, and principally in plantations of C. canephora. Only phenotype E1 of M. exigua was detected, distributed in 23.8% of the properties and only in plantations of C. arabica. M. exigua was found in all the municipalities sampled in the southern region of the state and in 66.6% of the municipalities of the Serrana region. M. paranaensis, phenotype P1, was detected only in the northern region, and in 100% of the properties sampled in Baixo Guandu municipality. The study was complemented by sampling in the Zona da Mata region of Minas Gerais state, where only M. exigua, phenotype E1, was found in C. arabica plantations. M. exigua was detected in all the sampled municipalities except Paula Cândido. The populations were subjected to differential host plant tests to determine the physiological races. Races 1 and 2 of M. incognita and M. exigua were found in Espírito Santo state, and race 2 of M. exigua in Zona da Mata in Minas Gerais state. The presence of the three most important root-knot nematodes in coffee plants in Espírito Santo state indicates the need to establish measures that will contain their dissemination.     

Chloropicrin Soil Fumigation Reduces <Emphasis Type="Italic">Spongospora subterranea</Emphasis> Soil Inoculum Levels but Does Not Control Powdery Scab Disease on Roots and Tubers of Potato

The effect of chloropicrin fumigation on the soil populations of Spongospora subterranea and the development of powdery scab, formation of root galls and tuber yield was investigated in seven field trials conducted in Minnesota and North Dakota. Sixteen potato cultivars, with different levels of susceptibility to disease on roots and tubers, were planted in plots treated with chloropicrin at rates ranging from zero to 201.8 kg a.i. ha^?1. The amount of S. subterranea DNA in soil was determined using qPCR. Bioassays were conducted to further assess the effect of chloropicrin fumigation on root colonization by S. subterranea in two potato cultivars with contrasting disease susceptibility. In the field, chloropicrin applied at rates between 70.1 to 201.8 kg a.i. ha^?1 significantly decreased S. subterranea initial inoculum in soil but increased the amount of disease observed on roots and tubers of susceptible cultivars. The effect of increasing disease was confirmed in controlled conditions experiments. Although the amount of S. subterranea DNA in roots of bioassay plants increased with increasing chloropicrin rates, it remained similar among potato cultivars. Chloropicrin fumigation significantly increased tuber yield which in cultivars such as Shepody and Umatilla Russet were associated with the amount root galls (r = 0.30; P < 0.03). Results of these studies contradict earlier reports on the use of chloropicrin fumigation for the control of powdery scab. Factors other than inoculum level, such as environmental conditions that affect inoculum efficiency and host susceptibility, may be significant contributors to the development of powdery scab and root gall formation.     

Biocontrol of bacterial wilt of potato with an avirulent strain ofpseudomonas solanacearum: interactions with root-knot nematodes

Field tests for biological control of bacterial wilt of potatoes, caused byPseudomonas solanacearum, were conducted in Florida in 1984 and 1985. Potato seedpieces of tolerant (Ontario) and susceptible (Pungo and/or Atlantic) cultivars were immersed in a suspension of the avirulent, non-bacteriocinogenic strain ofP. solanacearum, B82, prior to planting in naturally-infested soil. Relatively low populations ofP solanacearum (2 to 4 log cfu/g soil) and root-knot nematodes (0 to 33 larvae ofMeloidogyne incognita/100 cm³ soil) were present in the soil before planting. In 1984, there was approximately a 50% reduction in incidence of bacterial wilt and brown rot in B82-treated plants of cultivar Atlantic, as compared with controls. With cultivar Ontario, on the other hand, disease incidence was very low and differences between treatments were not significant. In 1985, the incidence of bacterial wilt was low in all plots. Treatment with strain B82 reduced incidence of tuber brown rot in all cultivars, but differences between treatments were not significant at P = 0.05. There was a significant correlation, however, between incidence of wilt and root-knot nematode populations in the soil. Therefore, the possibility that control of bacterial wilt in 1984 was indirect,i e, via an effect of B82 treatment on root infection byM incognita, was examined. Greenhouse tests were conducted to determine the potential effect of seedpiece treatment with strain B82 on nematode infection of cultivars Atlantic and Ontario. In two out of three trials, the B82 treatment significantly reduced incidence of root knot. These results suggest that interactions of the biocontrol agent with root-knot nematodes may determine the effectiveness of a treatment designed for bacterial wilt control in the field.     

Management of Meloidogyne incognita in yam-based cropping systems with cover crops

The effect of intercropping cover crops was with yam was assessed for nematode management both in pot and field conditions in Nigeria. The cover crops were sown with yams in pots containing sterile soil and inoculated with 5000 eggs of Meloidogyne incognita. In the field, each cover crop was separately intercropped with yams inoculated with 10,000 nematodes. From both pot and field experiments, no nematode damage was observed on yam tubers that were intercropped with Aeschynomene histrix, Crotolaria juncea, and Tagetes erecta. Damage in intercropped tubers was reduced by 72.7% with Mucuna pruriens, Centrosema pubescens, and Pueraria phaseoloides and by 58.3% with Stylosanthes guianensis compared to yams planted without cover crops; they also had lower (p ≤ 0.05) nematode populations. Cajanus cajan, Lablab purpureus and Vigna unguiculata however, supported high nematode populations and led to nematode damage in intercropped yams. Tubers from M. incognita-inoculated plants were more damaged and lost 42% more weight (p ≤ 0.05) following three months of storage than tubers from uninoculated plants. Intercropping yams with selected cover crops can be useful in managing M. incognita without reducing yam yields.     

Cultivation offers effective management of subterranean clover damping‐off and root disease

Soil cultivation studies involving subterranean clover pastures were undertaken utilizing field cores from five farms and two in‐field trials. Tap and lateral root disease in cores was less (p < .001) severe and root and shoot weights greater (p < .001) following simulated cultivation. Germination and severity of root disease were both affected (p < .005) by three‐way interactions with cultivation, cultivar and field site. Cultivation in cores suppressed tap root disease for cultivars Meteora and Riverina across the five sites and Seaton Park for two sites. In‐field trials confirmed cultivation reduces root disease severity and increases germination and plant productivity. The best in‐field treatment was cultivation + fumigation that reduced (p < .05) tap and lateral root disease and increased nodulation and root and shoot weights for Riverina, Seaton Park and Woogenellup. Cultivation + fumigation also increased (p < .05) germination for Woogenellup and Seaton Park. There were negative correlations (all p < .001) between tap and lateral root disease with nodulation (R² = .85, R² = .58, respectively); tap root disease with root and shoot weight (R² = .58, R² = .854, respectively); and lateral root disease with root and shoot weight (R² = .83, R² = .64 respectively). This study highlights the close relationship between severe root disease and reduced nodulation, likely explaining much of the widespread poor nodulation in subterranean clover pastures. This study confirms that damping‐off and root disease can be mitigated by cultivation, offering producers flexibility in disease management, especially where autumn–winter feed shortages occur on a regular basis.     

Nemato-toxic potential of Betel (Piper betle L.) (Piperaceae) leaf

In vitro and in-planta trials were conducted to determine the nemato-toxic potential of aqueous leaf extracts of Betel (Piper betle L.) against the root-knot nematode species, Meloidogyne incognita, at five concentrations, standard extract (S) [1:2 (w/v] and its four dilutions, 20%, 40%, 60%. 80% of S. In the in vitro trials, second-stage juveniles (J₂s) and eggs of M. incognita were directly exposed to the extract while in the in-planta trials, the effect of the extract was evaluated as a root dip treatment using tomato plants. The findings indicated that the extract was lethal to J₂s but also inhibited egg hatch. The seedling dip treatments reduced root infestations in terms of gall formation, egg production and J₂ population densities in soil while simultaneously enhancing the growth of tomato plants. All these effects varied in a dose-dependent manner. Based on the LC₅₀ value, the eggs were found to be less sensitive to the extract than J₂s. One hundred percent of mortality of J₂s was recorded at four concentration levels, 40%, 60% and 80% of S and S, while 100% egg inhibition was only recorded at S. The highest reductions in gall formation (i.e., number of galls/root system), egg production and J₂ population were all recorded at S as 83%, 87% and 84% compared to the untreated water control. The maximum growth enhancement, which was 235% of the control, was detected in the root length of treated tomato plants. It appears the aqueous leaf extract of P. betle shows promise as a sustainable eco-friendly nematicide for the management of M. incognita.     

Flame soil disinfestation: A novel,promising, non-chemical method to control soilborne nematodes,fungal and bacterial pathogens in China

Flame soil disinfestation (FSD) is a novel, promising non-chemical method to control soilborne nematodes, fungal and bacterial pathogens in China. The efficacy of FSD on soilborne nematodes, fungal and bacterial pathogens was evaluated during two field trials. The field trials revealed that the treatment with FSD once (FSD1) and treatment with FSD twice (FSD2) sharply reduced the total number of soilborne nematodes (>95%) and completely controlled Meloidogyne incognita in the soil. Both FSD1 and FSD2 also provided promising efficacy against soilborne Fusarium oxysporum (>44%), Phytophthora spp. (>47%) and Ralstonia solanacearum (>67%) on media. However, there was no significant difference between FSD1 and FSD2 in controlling soilborne nematodes, fungal and bacterial pathogens (P = 0.05). Currently, there are two challenges to distribute the technology in China: its high cost and relatively low efficiency because of the low speed of application compared with soil chemical fumigation. Despite the drawbacks, FSD is still promising in organic agriculture for controlling soilborne nematodes, fungal and bacterial pathogens.     

Root-knot and reniform nematode reproduction on kenaf and sunn hemp compared with that on nematode resistant and susceptible cotton

Kenaf (Hibiscus cannabinus L.) and sunn hemp (Crotalaria juncea L.) are industrial fiber crops that are being grown increasingly in areas where cotton (Gossypium hirsutum L.) is the major crop. The nematodes causing most cotton yield losses in the USA are the southern root-knot nematode, Meloidogyne incognita (Kofoid and White) Chitwood, and the reniform nematode, Rotylenchulus reniformis Linford and Oliveira. For nematode management, knowledge about the comparative effects of kenaf, sunn hemp, and cotton on nematode population densities in the soil is important. The objective of this study was to clarify the host status of kenaf and sunn hemp to M. incognita and R. reniformis by directly comparing reproduction of each nematode on each crop with the same nematode's reproduction on nematode-resistant and nematode-susceptible cotton. Comparisons were made in six experiments under growth chamber, microplot, and field conditions. The results showed that sunn hemp is resistant, but not equally resistant to M. incognita and R. reniformis. Reproduction of R. reniformis on sunn hemp was nearly undetectable, whereas reproduction of M. incognita was greater than on resistant cotton (Auburn 623, Auburn 634) and up to 20% of that on susceptible cotton (Deltapine 16, Deltapine 50, or Deltapine 5409). Kenaf was confirmed to elevate populations of M. incognita to levels that devestate cotton, but found to support substantially less reproduction by R. reniformis than cotton does. Two populations of R. reniformis differed substantially in reproduction on kenaf even though the same populations reproduced similarly on cotton. In conclusion, distinguishing M. incognita from R. reniformis is essential to the management of nematodes in rotations involving kenaf, sunn hemp and cotton.     

Histopathological changes induced by Meloidogyne incognita in some ornamental plants

Histopathological changes induced by the root-knot nematode (Meloidogyne incognita) in five ornamental plants, specifically, Calendula (Calendula officinalis), Centaurea (Centaurea montana), Papaver (Papaver somniferum), Chrysanthemum (Chrysanthemum morifolium) and Dianthus (Dianthus caryophyllus), were investigated. Based on the galling index (GI), Centaurea was classified as susceptible, Calendula as moderately susceptible, and Papaver as moderately resistant, while Chrysanthemum and Dianthus were highly resistant to M. incognita infection. The histopathology of the galled roots of Centaurea, Calendula, and Papaver during later stages of infection showed that nematodes were localized entirely within the cortex and generally oriented horizontally to the vascular cylinder. Most of the females were mature, and a few of them were associated with egg masses. Giant cells with a variation in cell sizes were observed in the galled roots of all three of the plant species and exhibited a granular cytoplasm and hypertrophied nuclei as a typical reaction to nematode feeding.     

Rapid detection of Meloidogyne spp. by LAMP assay in soil and roots

Loop-mediated isothermal amplification (LAMP), a novel DNA amplification technique, has been used to detect a variety of pathogens including viruses, fungi, bacteria and parasites. However, diagnosis of sedentary plant-parasitic nematode (PPN) species has not yet been attempted. In this study, we developed a universal LAMP set (RKN-LAMP) for the diagnosis of four common Meloidogyne species (Meloidogyne incognita, M. arenaria, M. javanica and M. hapla), and M. incognita-specific LAMP set (Mi-LAMP). In both assays, a typical ladder-like pattern on gel electrophoresis was observed in all positive samples but not in the negative controls. Amplification products were further confirmed using restriction analysis of the Hpa II enzyme, detection by visual inspection using SYBR Green I and the lateral flow dipstick (LFD) assay. The two LAMP sets were specifically able to detect four common Meloidogyne species and M. incognita populations having several different geographical origins and pathotypes. No cross reaction with DNA of other PPNs was observed. Sensitivity of the RKN- and Mi-LAMP was 10 and 100 fg of pure genome DNA respectively. Both LAMP sets could also amplified crude DNA isolated from the galled root tissue and from soil containing juveniles of M. incognita. The RKN- and Mi-LAMP sets offer the advantages of simplicity, rapidity and cost effectiveness. Both LAMP sets will be instrumental for the diagnosis of Meloidogyne spp. by local extension and regulatory personnel.