Control of root-knot nematode,Meloidogyne incognita and reniform nematode,Rotylenchulus reniformis on pigeonpea and linseed

Abstract

Pigeonpea (Cajanus cajan) and linseed (Linum usitatissimum) are susceptible to Meloidogyne incognita and Rotylenchulus reniformis nematodes. Reduction in different growth parameters (length and weight of plant, number of pods), bulk density of pigeonpea stem, oil content of linseed, chlorophyll content of leaf and water absorption of roots caused by M. incognita and R. reniformis were statistically significant. Similar effects were also observed in plants raised from seeds soaked in different concentrations of water soluble fractions (WSF) of rice polish and pyridoxine solutions, however, the reductions were of a comparatively lesser extent. Higher concentrations of the solutions were more effective when compared to lower ones and pyridoxine was more beneficial than WSF for improving plant growth and reducing disease incidence.     

A greenhouse technique to screen pigeonpea for resistance to Heterodera cajani*

Heterodera cajani is an important nematode pest of pigeonpea in India and a simple and reliable greenhouse procedure has been developed to screen pigeonpea genotypes for resistance to it. In pot experiments, white cysts of H. cajani were counted on the roots of the susceptible genotype ICPL 87 at 15, 30 and 45 days after seedling emergence in soils infested with different levels of H. cajani. The seedlings were rated for the number of white cysts per root system on a one (highly resistant, no cysts) to nine (highly susceptible, more than 30 cysts) scale. White cysts were not easy to see on wet roots but were clearly visible on slightly dried roots. Cyst counts and ratings were more uniform when roots of 30 day old seedlings were evaluated than when 15 or 45 day old seedlings were examined. Effects of different H. cajani infestation levels on the ratings were not significant although the use of higher inoculum densities (16 to 27 eggs and juveniles/cm³ soil) was effective in reducing variability. This procedure was used to screen 60 pigeonpea genotypes and all of them were rated seven or nine. Ten accessions of Atylosia spp. and Rhynchosia spp. were rated three.     

Tolerance to reniform nematode (Rotylenchulus reniformis) race A in pigeonpea (Cajanus cajan) genotypes

The reniform nematode (Rotylenchulus reniformis) is an important pathogen of pigeonpea (Cajanus cajan). Forty‐six medium maturity (mature in 151–200 days at Patancheru, India) pigeonpea genotypes were evaluated for resistance and tolerance to the reniform nematode in greenhouse and field tests, over the period 1990–97. Each genotype was screened for number of nematode egg masses on a 1 (no egg mass = highly resistant) to 9 (> 50 egg masses = highly susceptible) scale. Plant biomass production in carbofurantreated plots was compared with that in non‐treated plots in a field naturally infested with R. reniformis. Pigeonpea genotypes C 11, ICPL 87119 and ICPL 270 were used as nematode susceptible checks. Genotypes with good plant growth, both in nematode‐free and nematode‐infested plots, were identified as tolerant and evaluated for plant growth and yield for at least three years. All the tested genotypes were susceptible (7 and 9 egg mass score). Single‐plant‐selections, based on plant vigour and yield, were made from genotypes showing tolerance to nematode infection. The level of tolerance was enhanced by plant‐to‐progeny row selection for plant vigour and seed yield in a nematode‐sick field for at least three years. The most promising nematode tolerant genotypes produced significantly greater yield and biomass than the locally grown pigeonpea cultivars in fields naturally infested with R. reniformis at two locations. Pigeonpea landraces are considered to be the most likely sources of tolerance to the nematode. These reniform nematode tolerant lines represent new germplasm and they are available in the genebank of pigeonpea at ICRISAT bearing accession numbers ICP 16329, ICP 16330, ICP 16331, ICP 16332, and ICP 16333.     

Host status of some broad bean,Vicia faba L., selections to reniform nematode,Rotylenchulus reniformis,under greenhouse conditions in Egypt

Twenty broad bean selections were screened for their relative susceptibility to the reniform nematode, Rotylenchulus reniformis, under greenhouse conditions at 20?±?5?°C. The degree of susceptibility was rated based on number of females per root system of each selection. Hence, G 3, L 47, L 49, L 52, L 57, L 71, L 92, L 99 and L 375 selections were graded as resistant hosts against the nematode infection and sex selections (G 429, L 13, L 16, L 24, L 31 and L 46) were categorised as moderately resistant. However, only the selection L 9 out of 20 selections was ranked as susceptible and four selections, namely L 50, L 101, L 241 and L 348, were rated as highly susceptible against R. reniformis infection. It was observed that reproduction of the nematode was favoured on highly susceptible and susceptible selections but inhibited on resistant ones. Therefore, all tested selections showed great variability in their reaction to the nematode infection according to the host type. Also, various plant growth parameters were discussed. Finally, the differences among the tested selections should serve as an informative resource for plant breeders and cropping systems to limit the loss due to the nematode infection.     

Studies on the pathogenicity of three host races of Rotylenchulus reniformis on castor

Three races (race 2, 3 and 4) of Rotylenchulus reniformis were identified using host differential from 20 populations collected from Aligarh, India. Pathogenicity tests were conducted using 250, 500, 1000, 2000, 4000 and 8000 immature females/kg soil of three races of R. reniformis on castor. The threshold inoculum levels of Race-2, Race-3 and Race-4 on castor were 500, 1000 and 2000 immature females/kg soil, respectively. Out of these three races present in Aligarh district, Race-2 was found more pathogenic followed by Race-3 and Race-4. Infected plants with all the three races of reniform nematode showed stunting, necrosis, leaf shedding and growth reduction at and above the inoculum threshold level of each race. Reduction in castor growth was directly proportional to the inoculum level of R.reniformis. The rate of nematode multiplication was density dependent.     

Identification of Rotylenchulus reniformis Resistant Glycine Lines

Identification of resistance to reniform nematode (Rotylenchulus reniformis) is the first step in developing resistant soybean (Glycine max) cultivars that will benefit growers in the mid-South region of the United States. This study was conducted to identify soybean (G. max and G. soja) lines with resistance to this pathogen. Sixty-one wild and domestic soybean lines were evaluated in replicated growth chamber tests. Six previously untested soybean lines with useful levels of resistance to reniform nematode were identified in both initial screening and subsequent confirmation tests: released germplasm lines DS4-SCN05 (PI 656647) and DS-880 (PI 659348); accession PI 567516 C; and breeding lines DS97-84-1, 02011-126-1-1-2-1 and 02011-126-1-1-5-1. Eleven previously untested moderately susceptible or susceptible lines were also identified: released germplasm lines D68-0099 (PI 573285) and LG01-5087-5; accessions PI 200538, PI 416937, PI 423941, PI 437697, PI 467312, PI 468916, PI 594692, and PI 603751 A; and cultivar Stafford (PI 508269). Results of previously tested lines evaluated in the current study agreed with published reports 69.6% of the time for resistant lines and 87.5% of the time for susceptible lines. Soybean breeders may benefit from incorporating the newly identified resistant lines into their breeding programs.     

Evaluation of Meloidogyne incognita and Rotylenchulus reniformis nematode-resistant cotton cultivars with supplemental Corteva Agriscience nematicides

Meloidogyne incognita- and Rotylenchulus reniformis-resistant new cotton cultivars have recently become available, giving growers a new option in nematode management. The objectives of this study were: (i) to determine the yield potential of the new cultivars PHY 360 W3FE (M. incognita-resistant) and PHY 332 W3FE (R. reniformis-resistant) in nematode-infested fields and (ii) to evaluate the effects of combining the nematicides Reklemel (fluazaindolizine), Vydate C-LV (oxamyl), and the seed treatment BIO^ST Nematicide 100 (heat killed Burkholderia rinojenses and its non-living spent fermentation media) with resistant cotton cultivars on nematode population levels and lint yield. Field experiments in 2020 and 2021 indicated M. incognita population levels were 73% lower on PHY 360 W3FE (R) and 80% lower for R. reniformis on the PHY 332 W3FE (R) at 40 days after planting. Nematode eggs per gram of root were further reduced an average of 86% after the addition of Reklemel and Vydate C-LV when averaging both cultivars over the two years. Tests with BIO^ST Nematicide 100 + Reklemel + Vydate C-LV (0.56 + 2.5 L/ha) in both M. incognita and R. reniformis fields produced higher lint yields. Overall, planting PHY 360 W3FE (R) and PHY 332 W3FE (R) improved yields an average of 364 kg/ha while limiting nematode population increases. The addition of the nematicides further increased yields 152 kg/ha of the nematode-resistant cultivars.     

Effect of Temperature on the Embryogenesis of Geographic Populations of Rotylenchulus reniformis

The effect of temperature on the embryonic development of three populations of reniform nematode (Rotylenchulus reniformis) from the southeastern United States was studied. The development of eggs from single-cell stage to eclosion of second-stage juvenile was monitored at 20, 25, 30, and 35°C. All populations completed embryogenesis in 7 days at 25°C. The greatest differences among populations in time to completion of embryogenesis were observed at 20 and 35°C. Results at the intermediate temperatures (25 and 30°C) were similar for the three populations. The optimal temperature for embryogenesis was calculated to be 31.4°C for the population from Alabama, 28.4°C for the one from Mississippi, and 37.5°C for the one from South Carolina.     

Postinfection Development of Rotylenchulus reniformis on Resistant Gossypium barbadense Accessions

The reniform nematode (Rotylenchulus reniformis) causes significant cotton (Gossypium hirsutum) losses in the southeastern United States. The research objective was to describe the effects of two resistant G. barbadense lines (cultivar TX 110 and accession GB 713) on development and fecundity of reniform nematode. Nematode development and fecundity were evaluated on the resistant lines and susceptible G. hirsutum cultivar Deltapine 16 in three repeated growth chamber experiments. Nematode development on roots early and late in the infection cycle was measured at set intervals from 1 to 25 d after inoculation (DAI) and genotypes were compared based on the number of nematodes in four developmental stages (vermiform, swelling, reniform, and gravid). At 15, 20, and 25 DAI, egg production by individual females parasitizing each genotype was measured. Unique reniform nematode developmental patterns were noted on each of the cotton genotypes. During the early stages of infection, infection and development occurred 1 d faster on susceptible cotton than on the resistant genotypes. Later, progression to the reniform and gravid stages of development occurred first on the susceptible genotype, followed by G. barbadense cultivar TX 110, and finally G. barbadense accession GB 713. Egg production by individual nematodes infecting the three genotypes was similar. This study corroborates delayed development previously reported on G. barbadense cultivar TX 110 and is the first report of delayed infection and development associated with G. barbadense accession GB 713. The different developmental patterns in the resistant genotypes suggest that unique or additional loci may confer resistance in these two lines.     

Effects of Acibenzolar-S-Methyl Application to Rotylenchulus reniformis and Meloidogyne javanica

Effects of acibenzolar-s-methyl, an inducer of systemic acquired resistance in plants, on Rotylenchulus reniformis and Meloidogyne javanica in vitro and in vivo were determined. A single foliar application of acibenzolar at 50 mg/liter (5 ml of solution per plant) to 7-day-old cowpea or soybean seedlings decreased R. reniformis and M. javanica egg production by 50% 30 days after inoculation. The mechanism of acibenzolar on plant-parasitic nematodes was then investigated. Acibenzolar at 50 to 200 mg/liter did not affect movement of R. reniformis and M. javanica or penetration of second-stage juveniles (J2) of M. javanica on cowpea. However, M. javanica development was slowed and fecundity was reduced in plants treated with acibenzolar. On average, 50% of J2 that penetrated acibenzolar-treated cowpeas developed into mature females with eggs, whereas the other 50% exhibited arrested development. The number of eggs per egg mass was 450 in water-treated cowpeas, whereas the number declined to 250 in acibenzolar-treated plants. Acibenzolar may be responsible for stimulating the plants to express some resistance to the nematodes.     

Extraction of Root-associated Meloidogyne incognita and Rotylenchulus reniformis

A technique based on physical maceration of root tissue was developed to extract vermiform and swollen stages of Meloidogyne incognita and Rotylenchulus reniformis. Experiments conducted on soybean and tomato evaluated the efficiency of method (stir, grind), NaOC1 concentration (0%, 0.5%), and duration (lx, 2x) on extraction of nematodes and eggs from 60-day-old populations. Root-associated populations of R. reniformis were considerably lower than those of M. incognita, so development of the method focused on the latter. Grinding liberated more nematodes than stirring, but the reverse was true for egg extraction. Among grinding treatments, a duration of 10 seconds in 0.5% NaOCl provided the most efficient extraction of nematodes and eggs. Among stirring treatments, a duration of 10 minutes in 0.5% NaOCl provided the most efficient extraction of eggs. These techniques were compared on soybean roots 30 days older than those on which the procedures were first evaluated, with consistent results.     

Studies on inheritance of resistance and allelic relationships for strain 2 of pigeonpea sterility mosaic pathogen

Inheritance of resistance and allelic relationships were studied in three resistant pigeonpea sources for strain 2 of sterility mosaic pathogen. The resistant genotypes (ICP 7035, ICP 7349 and ICP 8850) were crossed with susceptible genotypes (BDN1 and LRG30) to determine the inheritance of resistance. The resistant and susceptible genotypes were also crossed among themselves to obtain information on their allelic relationships. Parents, F₁ and F₂ generations were sown in pots and screened using infector-hedge technique. Observations in parents, F₁ and F₂ generations, indicated dominance of resistance in certain crosses and the dominance of susceptibility in others. Disease reaction appeared to be governed by two independent non-allelic genes, with at least three multiple alleles, at one of the loci.     

Resistance to cyst nematode (Heterodera cajani) in pigeonpea cultivars and in wild relatives of Cajanus*

Heterodera cajani is an important nematode pest of pigeonpea in India. Evaluation of 58 pigeonpea cultivars and 61 accessions of Cajanus acutifolius, C. cajanifolius, C. grandifolius, C. lanceolatus, C. lineatus, C. mollis, C. pla-tycarpus, C. reticulatus, C. scarabaeoides, C. sericeus, C. volubilis, Flemingia macrophylla, F. stricta, F. strobilifera, Rhynchosia aurea, R. bracteata, R. cana, R. densiflora, R. minima, R. rothii, R. suaveolens and R. sublobata revealed that the tested pigeonpea cultivars lacked resistance to H. cajani. Eight accessions of wild relatives were resistant and 20 accessions were moderately resistant. Based on the white cyst number on roots and low plant-to-plant variation, two accessions of C. scarabaeoides (ICPWs 111 and 128), three accessions of Flemingia spp. (ICPWs 194, 202 and 203), and one accession each of R. rothii (ICPW 257), R. densiflora (ICPW 224), and R. aurea (ICPW 210) were identified as resistant and promising for use in intergeneric hybridisation programmes.     

Two Simple Methods for the Collection of Individual Life Stages of Reniform Nematode,Rotylenchulus reniformis

The sedentary semi-endoparasitic nematode Rotylenchulus reniformis, the reniform nematode, is a serious pest of cotton and soybean in the United States. In recent years, interest in the molecular biology of the interaction between R. reniformis and its plant hosts has increased; however, the unusual life cycle of R. reniformis presents a unique set of challenges to researchers who wish to study the developmental expression of a particular nematode gene or evaluate life stage–specific effects of a specific treatment such as RNA-interference or a potential nematicide. In this report, we describe a simple method to collect R. reniformis juvenile and vermiform adult life stages under in vitro conditions and a second method to collect viable parasitic sedentary females from host plant roots. Rotylenchulus reniformis eggs were hatched over a Baermann funnel and the resultant second-stage juveniles incubated in petri plates containing sterile water at 30°C. Nematode development was monitored through the appearance of fourth-stage juveniles and specific time-points at which each developmental stage predominated were determined. Viable parasitic sedentary females were collected from infected roots using a second method that combined blending, sieving, and sucrose flotation. Rotylenchulus reniformis life stages collected with these methods can be used for nucleic acid or protein extraction or other experimental purposes that rely on life stage–specific data.     

Suppression of Rotylenchulus reniformis 122-cm Deep Endorses Resistance Introgression in Gossypium

Nine sources of resistance to Rotylenchulus reniformis in Gossypium (cotton) were tested by measuring population density (Pf) and root-length density 0 to 122 cm deep. A Pf in the plow layer less than the autumn sample treatment threshold used by consultants was considered the minimum criterion for acceptable resistance, regardless of population density at planting (Pi). Other criteria were ample roots and a Pf lower than on the susceptible control, as in pot studies. In a Texas field in 2001 and 2002, no resistant accessions had Pf less than the control but all did in microplots into which nematodes from Louisiana were introduced. An environmental chamber experiment ruled out nematode genetic variance and implicated unknown soil factors. Pf in field experiments in Louisiana, Mississippi, and Alabama were below threshold for zero, six and four of the accessions and above threshold in the control. Gossypium arboreum A2–87 and G. barbadense GB-713 were the most resistant accessions. Results indicate that cultivars developed from these sources will suppress R. reniformis populations but less than in pots in a single season.     

Effects of Soil Solarization on Rotylenchulus reniformis in the Lower Rio Grande Valley of Texas

Soil solarization was evaluated for control of Rotylenchulus reniformis in the lower Rio Grande Valley of Texas. In field experiments, solarization significantly reduced soil nematode population densities 0-15 cm deep and increased yields of lettuce and cowpea. The length of time required for 90% mortality of nematodes in soil heated under controlled conditions in the laboratory varied from 25 hours to less than 1 hour between 41 and 47 C. Daily exposures of nematode-infested soil to lethal temperatures for sublethal time periods had a cumulative lethal effect. In water, vermiform stages required up to 10 days to recover from sublethal thermal stress. Eggs were similar to juveniles in their sensitivity to high temperatures. Lethal time-temperatures under controlled conditions were in general agreement with field results.     

Vertical Distribution of Rotylenchulus reniformis in Cotton Fields

The possible impact of Rotylenchulus reniformis below plow depth was evaluated by measuring the vertical distribution of R. reniformis and soil texture in 20 symptomatic fields on 17 farms across six states. The mean nematode population density per field, 0 to 122 cm deep, ranged from 0.4 to 63 nematodes/g soil, and in 15 fields more than half of the R. reniformis present were below 30.5 cm, which is the greatest depth usually plowed by farmers or sampled by consultants. In 11 fields measured, root density was greatest in the top 15 cm of soil; however, roots consistently penetrated 92 to 122 cm deep by midseason, and in five fields in Texas and Louisiana the ratio of nematodes to root-length density within soil increased with depth. Repeated sampling during the year in Texas indicated that up to 20% of the nematodes in soil below 60 cm in the fall survived the winter. Differences between Baermann funnel and sugar flotation extraction methods were not important when compared with field-to-field differences in nematode populations and field-specific vertical distribution patterns. The results support the interpretation that R. reniformis below plow depth can significantly impact diagnosis and treatment of cotton fields infested with R. reniformis.     

Comparisons of Female and Egg Assays to Identify Rotylenchulus reniformis Resistance in Cotton

More plants can be screened for reniform nematode resistance each year if the time involved can be shortened. In this study, the hypothesis that female counts are as efficient as egg counts in identifying resistant genotypes was tested. In two greenhouse experiments Gossypium genotypes which varied from resistant to susceptible to reniform nematode (Rotylenchulus reniformis) were compared to a susceptible control cultivar. Infested field soil served as the inoculum source for the first experiment, and vermiform stages extracted from greenhouse cultures were used to infest soil in the second experiment. Six replicates of each genotype were harvested 25 d after planting and swollen females were counted. The remaining plants were harvested 35 d after planting and eggs extracted from the roots were counted. Processing and counting times recorded in the first experiment were similar for both assessment methods, but 10 additional days were required for egg-based assessment. Contrast analyses showed that assessments based on females per gram of root were equivalent to assessments based on eggs per gram of root for the five genotypes tested in the first experiment and for an expanded set of 13 genotypes tested in the second experiment. The results indicated that either life stage can be used to screen for resistance.     

Sensitivity of Meloidogyne incognita and Rotylenchulus reniformis to Abamectin

Avermectins are macrocyclic lactones produced by Streptomyces avermitilis. Abamectin is a blend of B_1a and B_1b avermectins that is being used as a seed treatment to control plant-parasitic nematodes on cotton and some vegetable crops. No LD₅₀ values, data on nematode recovery following brief exposure, or effects of sublethal concentrations on infectivity of the plant-parasitic nematodes Meloidogyne incognita or Rotylenchulus reniformis are available. Using an assay of nematode mobility, LD₅₀ values of 1.56 μg/ml and 32.9 μg/ml were calculated based on 2 hr exposure for M. incognita and R. reniformis, respectively. There was no recovery of either nematode after exposure for 1 hr. Mortality of M. incognita continued to increase following a 1 hr exposure, whereas R. reniformis mortality remained unchanged at 24 hr after the nematodes were removed from the abamectin solution. Sublethal concentrations of 1.56 to 0.39 μg/ml for M. incognita and 32.9 to 8.2 μg/ml for R. reniformis reduced infectivity of each nematode on tomato roots. The toxicity of abamectin to these nematodes was comparable to that of aldicarb.     

Sensitivity of Meloidogyne incognita and Rotylenchulus reniformis to Fluopyram

Fluopyram is a succinate dehydrogenase inhibitor (SDHI) fungicide that is being evaluated as a seed treatment and in-furrow spray at planting on row crops for management of fungal diseases and its effect on plant-parasitic nematodes. Currently, there are no data on nematode toxicity, nematode recovery, or effects on nematode infection for Meloidogyne incognita or Rotylenchulus reniformis after exposure to low concentrations of fluopyram. Nematode toxicity and recovery experiments were conducted in aqueous solutions of fluopyram, while root infection assays were conducted on tomato. Nematode paralysis was observed after 2 hr of exposure at 1.0 µg/ml fluopyram for both nematode species. Using an assay of nematode motility, 2-hr EC₅₀ values of 5.18 and 12.99 µg/ml fluopyram were calculated for M. incognita and R. reniformis, respectively. Nematode recovery in motility was greater than 50% for M. incognita and R. reniformis 24 hr after nematodes were rinsed and removed from a 1-hr treatment of 5.18 and 12.99 µg/ml fluopyram, respectively. Nematode infection of tomato roots was reduced and inversely proportional to 1-hr treatments with water solutions of fluopyram at low concentrations, which ranged from 1.3 to 5.2 µg/ml for M. incognita and 3.3 to 13.0 µg/ml for R. reniformis. Though fluopyram is nematistatic, low concentrations of the fungicide were effective at reducing the ability of both nematode species to infect tomato roots.