DNA sequence analyses of the nuclear ribosomal ITS1 region of the ribosomal DNA and D1-D3 expansion segments of the 28S gene were conducted to characterize the genetic variation of six amphimictic Hoplolaimus species, including H. magnistylus, H. concaudajuvencus, H. galeatus, Hoplolaimus sp. 1, Hoplolaimus sp. 2 and Hoplolaimus sp. 3, and two closely related parthenogenetic species, H. columbus and H. seinhorsti. PCR amplifications of the combined D1-D3 expansion segments and the ITS1 region each yielded one distinct amplicon. In the D1-D3 region, there was no nucleotide sequence variation between populations of H. columbus, H. magnistylus, Hoplolaimus sp. 2 and Hoplolaimus sp. 3, whereas the ITS1 sequences had nucleotide variation among species. We detected conserved ITS1 regions located at the 3’ and 5’ end of ITS1 and also in the middle of the ITS1 among Hoplolaimus species. These regions were compared with sequences of distantly related Heterodera and Globedera. PCR-RFLP and sequence analysis of ITS1 and 28S PCR products revealed that several haplotypes existed in the same genome of H. columbus, H. magnistylus, H. seinhorsti, H. concaudajuvencus and Hoplolaimus sp. 1. Maximum likelihood and maximum parsimony analysis using the combined ITS1 and D1-D3 expansion segment sequences always produced trees with similar topology; H. columbus and H. seinhorsti grouped in one clade and the other six species (H. galeatus, H. concaudajuvencus, H. magnistylus, Hoplolaimus sp. 1, Hoplolaimus sp. 2, Hoplolaimus sp. 3) grouped in another. Molecular analysis supports morphological schemes for this genus to be divided into two groups based on several phenotypic traits derived from morphological evolution.
28S gene; clades; D1-D3 region genome; haplotypes; Hoplolaimus, lance; ITS1; nematode
DNA sequences of the D2-D3 expansion segments of the 28S gene of ribosomal DNA from 23 taxa of the subfamily Hoplolaiminae were obtained and aligned to infer phylogenetic relationships. The D2 and D3 expansion regions are G-C rich (59.2%), with up to 20.7% genetic divergence between Scutellonema brachyurum and Hoplolaimus concaudajuvencus. Molecular phylogenetic analysis using maximum likelihood and maximum parsimony was conducted using the D2-D3 sequence data. Of 558 characters, 254 characters (45.5%) were variable and 198 characters (35.4%) were parsimony informative. All phylogenetic methods produced a similar topology with two distinct clades: One clade consists of all Hoplolaimus species while the other clade consists of the rest of the studied Hoplolaiminae genera. This result suggests that Hoplolaimus is monophyletic. Another clade consisted of Aorolaimus, Helicotylenchus, Rotylenchus, and Scutellonema species. Phylogenetic analysis using the outgroup species Globodera rostocheinsis suggests that Hoplolaiminae is paraphyletic. In this study, the D2-D3 region had levels of DNA sequence divergence sufficient for phylogenetic analysis and delimiting species of Hoplolaiminae.
28S; analysis; Aorolaimus; clade; D2-D3; Helicotylenchus; Hoplolaiminae; Hoplolaimus; lance; nematode; phylogenetic; Rotylenchus; species; spiral; Scutellonema; taxonomy
Hoplolaimus concaudajuvencus n. sp., of the genus Hoplolaimus Daday, 1905, characterized by larval heteromorphism, is described and illustrated as recovered from ryegrass/bermudagrass golf green turf in Florida. Females and males are closely related to H. galeatus (Cobb, 1913) Thorne, 1935, but have longer stylets with more definitely tulip-shaped stylet knobs which anteriorly tend to close upon the stylet shaft more than in H. galeatus. First and second-stage larvae have a conically-pointed tail unlike any known species of the genus. Subsequent stages, including females, have rounded tails essentially similar to other species of the genus and males possess the typical hopolaimid tail and bursa. The first molt was found to occur within the egg.
Taxonomy; Hoplolaimus concaudajuvencus n. sp.; Morphology; Larval heteromorphism
Ten species of stylet-bearing nematodes were recovered in a survey of sycamore (Platanus occidentalis L. ) stands in Georgia. Helicotylenchus, Xiphinema, and Criconemoides were the genera found most frequently. Populations of Hoplolaimus galeatus, Scutellonema brachyurum, Helicotylenchus dihystera and H. pseudorobustus increased on greenhouse-grown sycamore, but Trichodorus christiei, Xiphinema americanum, Meloidogyne hapla, M. arenaria and M. incognita did not. Hoplolaimus galeatus and S. brachyurum are semi-endoparasites; H. dihystera and H. pseudorobustus are migratory endoparasites. Hoplolaimus galeatus caused extensive root necrosis and marked decrease of fresh weights of seedling roots and tops. Helicotylenchus dihystera and S. brachyurum produced only qualitatively different sparse and unhealthy root growth. Helicotylenchus pseudorobustus caused only a reduction in root surface area.
Helicotylenchus; Xiphinema; Hoplolaimus; Trichodorus; Meloidogyne
Treated and nontreated field plots were assayed, and the population density of each plant-parasitic nematode present was compared with crop growth and yield and with the population densities of other nematode species in the same plots. The strongest correlations between nematode population densities and growth responses occurred when soil assays for nematodes were made 55-73 days after planting. Belonolairnus longicaudatus was the most damaging parasite on peanut, Arachis hypogaea, as evidenced by high negative correlations between population densities and plant growth responses. Criconemoides ornatus, Meloidogyne hapla, Helicotylenchus dihystera, Trichodorus christie, Tylenchorhynchus claytoni, and Pratylenchus brachyurus were involved to varying degrees, depending on previous crop and initial densities of these nematodes. Hoplolaimus galeatus and Xiphinema americanum did not appear to affect crop response. The negative correlation of Trichodorus christiei to yield of soybean, Glycine max, was higher than that of Belonolairnus longicaudatus, although both contributed to yield losses. Similar correlation analyses showed that apparent antagonistic or synergistic population-density relationships among nematodes under field conditions depend on the time of sampling and the composition of the nematode community under study.
Belonolaimus longicaudatus and Hoplolaimus galeatus are considered among the most damaging pathogens of turfgrasses in Florida. However, the host status of seashore paspalum (Paspalum vaginatum) is unknown. Glasshouse experiments were performed in 2002 and 2003 to determine the tolerance of 'SeaIsle 1' seashore paspalum to a population of B. longicaudatus and a population of H. galeatus, and to compare to 'Tifdwarf' bermudagrass for differences. Both nematode species reproduced well on either grass, but only B. longicaudatus consistently reduced root growth as measured by root length. Belonolaimus longicaudatus reduced root growth (P ≤ 0.05) by 35% to 45% at 120 days after inoculation on both grasses. In 2003, higher inoculum levels of H. galeatus reduced root growth (P ≤ 0.05) by 19.4% in seashore paspalum and by 14% in bermudagrass after 60 and 120 days of exposure, respectively. Percentage reductions in root length caused by H. galeatus and B. longicaudatus indicated no differences between grass species, although Tifdwarf bermudagrass supported higher soil population densities of both nematodes than SeaIsle 1 seashore paspalum.
Belonolaimus longicaudatus; bermudagrass; Hoplolaimus galeatus; host status; lance nematode; Paspalum vaginatum; seashore paspalum; sting nematode; tolerance
The host-parasite relationships of 13 species of plant parasitic nematodes and five species of hardwoods native to the southeastern United States were tested on greenhouse-grown tree seedlings for 6-10 months. Criteria for parasitism were completion o f life cycle and population increase of nematodes. Belonolaimus longicaudatus, Helicotylenchus dihystera, Scutellonema brachyurum and Tylenchorhynchus claytoni parasitized and reproduced on three or more of the species tested. Hoplolaimus galeatus and Pratylenchus brachyurus parasitized two species, Trichodorus christiei and Criconemoides xenoplax parasitized only red maple. Meloidogyne javanica/Liriodendron tulipifera combination was the only positive root-knot nematode/hardwood host-parasite relationship. Hemicycliophora silvestris, Meloidogyne arenaria, M. incognita, and M. hapla were not parasites of the tree species tested.
Host-parasite relations; Liquidambar styraciflua; Acer rubrum; Liriodendron tulipifera; Platanus occidentalis; Populus heterophylla
During September 1990, 30 cotton fields in each of three Missouri counties were surveyed for plant-parasitic nematodes. Soil samples for nematode analysis consisted of a composite of 20 cores collected in a zig-zag pattern within a 1-ha block in each field. Cores were taken from within weed-free cotton rows. Nine genera of plant-parasitic nematodes were found (Rotylenchulus, Helicotylenchus, Hoplolaimus, Meloidogyne, Paratylenchus, Pratylenchus, Tylenchorhynchus, Heterodera, and Trichodorus), and five species were identified: Meloidogyne incognita, Rotylenchulus reniformis, Hoplolaimus galeatus, Pratylenchus vulnus, and P. scribneri. This is the first report of R. reniformis, H. galeatus, P. vulnus, and P. scribneri in Missouri cotton fields and the first report of R. reniformis and P. vulnus in Missouri. The known cotton pathogens M. incognita, R. reniformis, and H. galeatus were found in 30%, 3%, and 2% of the fields sampled, respectively. The correlation between sand content of the soil sample and the number of vermiform M. incognita in the sample was not significant, with r² = 0.13. Select fields where H. galeatus and R. reniformis were found in 1990 were sampled more intensely in 1991. The 1-ha block sampled in 1990 was sampled in 1991, along with three other 1-ha blocks uniformly distributed within the field. In addition, a 1-ha block was sampled in each of eight nearby fields, within 2 km of the first field. The nine plant-parasitic nematode genera identified in the 1990 survey were observed again in 1991. Within-field distribution of M. incognita, R. reniformis, and H. galeatus was not uniform. When M. incognita, R. reniformis, or H. galeatus were present in a field, the same species was found in 38%, 25%, or 50% of nearby fields, respectively.
cotton; Gossypium hirsutum; Meloidogyne incognita; Missouri; nematode; Rotylenchulus reniformis; Hoplolaimus galeatus; survey
Avermectin B₁, isazofos, and fenamiphos were evaluated in greenhouse experiments for efficacy against two common turfgrass parasites, Hoplolaimus galeatus and Tylenchorhynchus dubius. Treatments in all experiments were arranged in a completely randomized design and replicated four times. In the first experiment, avermectin B₁ at rates of 0.2 and 0.4 kg a.i./ha and isazofos at rates of 2.3 and 23 kg a.i./ha significantly reduced populations of both species of parasitic nematodes compared to controls at 14 and 28 days after treatment (P ≤ 0.01). In the second experiment, the greatest reductions in both nematode populations occurred at 28 and 56 days after treatment, where 23 kg a.i./ha of isazofos was applied (P ≤ 0.01). These reductions, however, were not different from reductions of H. galeatus at 28 and 56 days after treatment (P ≤ 0.01) or T. dubius at 56 days after treatment (P ≤ 0.01), where 0.2- and 0.4-kg a.i./ha rates of avermectin B₁ were mixed throughout the soil. In the third experiment, the greatest population reduction of H. galeatus was observed with a 0.4-kg a.i./ha treatment of avermectin B₁ at 56 days after treatment (P ≤ 0.05). T. dubius populations were reduced by the 0.4-kg a.i./ha rate of avermectin B₁ at 28 (P ≤ 0.01), 56 (P ≤ 0.05), and 70 (P ≤ 0.01) days after treatment. In the fourth and fifth experiments, avermectin B₁ at rates of 7.5 and 15.2 kg a.i./ha consistently reduced nematode populations compared to controls and performed as well or better than fenamiphos (P ≤ 0.01).
avermectin; chemical control; fenamiphos; Hoplolaimus galeatus; isazofos; nematode; Poa annua; turfgrass; Tylenchorhynchus dubius
Alyceclover (Alysicarpus spp.) is an annual, high-quality leguminous forage, suitable for production under tropical and subtropical climates where the husbandry of conventional leguminous forages, Trifolium spp., is uneconomical. The damage potential and reproduction of Belonolairaus longicaudatus and Hoplolaimus galeatus on alyceclover were studied under greenhouse conditions, using sand and sandy clay loam soil materials, respectively. Both nematode species reproduced on alyceclover, but only B. longicaudatus was pathogenic. Symptoms of B. longicaudatus damage were suppression of shoot yield, limited root system, stunting, incipient wilting, and occasional seedling mortality. In one experiment, the threshold-damage density was three nematodes/100 cm³ sand, whereas in the other experiment it was zero nematodes.
Alysicarpus spp.; damage threshold; lance nematode; leguminous forage; nematode; sting nematode; susceptible host; tolerant host
Results of surveys from 1978 to 1986 to estimate and identify nematodes in Arkansas soybean fields are presented. The seven most common nematode species in the fields were Heterodera glycines, Quinisulcius acutus, Pratylenchus scribneri, P. alleni, Helicotylenchus pseudorobustus, Tylenchorhynchus ewingi, and Xiphinema americanum. Other nematodes identified from these fields were Pratylenchus brachyurus, P. vulnus, P. zeae, Tylenchorhynchus canalis, T. goffarti, T. martini, Helicotylenchus dihystera, Scutellonema bradys, Xiphinema chambersi, X. rivesi, Hoplolaimus galeatus, H. magnistylus, Paratrichodorus minor (P. christiei), Paratylenchus projectus, P. tenuicaudatus, Criconemella macrodora, C. ornata, and Meiodorus hollisi.
Arkansas; crop lossestimate; Helicotylenchus pseudorobustus; Heterodera glycines; infestation estimate; Pratylenchus alleni; P. scribneri; Quinisulcius acutus; soybean; Tylenchorhynchus ewingi; Xiphinema americanum
A 2-year study was conducted on Merion Kentucky bluegrass turf (Poa pratensis) to identify potential relationships among seasonal population dynamics of nematodes, chemical applications, thatch, tillering, dollar spot caused by Sclerotinia homoeocarpa, clipping weight, and other factors. Numbers of Tylenchorhynchus maximus determined during June were inversely related to the wet weight of grass from May. One or more monthly counts of Paratylenchus hamatus, Criconemella rusium, and T. maximus negatively correlated with the numbers of spring tillers. Applications of benomyl, used for dollar spot control, decreased numbers of T. maximus and free-living nematodes, and this chemical was associated with acidification of the thatch. Hoplolaimus galeatus levels were associated with an estimated 8% increase in the severity of dollar spot.
Criconemella rusium; dollar spot; Hoplolaimus galeatus; Kentucky bluegrass; nematode; Paratylenchus hamatus; Poa pratensis; Sclerotinia homoeocarpa; turfgrass; Tylenchorhynchus maximus
A Pasteuria isolate associated with a population of the lance nematode Hoplolaimus galeatus was discovered in Peru. The infective propagules adhered to adult stages and juveniles and were found filling the bodies of males and females. The endospore and central core diameters measured 4.5 ± 0.4 pm and 1.9 ± 0.2 μm, respectively, which differed from those reported for other Pasteuria isolates found iu North America on the same host. Examinations of endospore ultrastructure with scanning electron microscopy showed the presence of a thin layer of parasporal fibers surrounding the central core, a thin reduced layer of parasporal fibers in contact with the host's cuticle, and a putative basal core ring.
biodiversity; biological control; endospore; Hoplolaimus galeatus; lance nematode; nematode; parasitism; Pasteuria penetrans; Peru
Examination of dispersional characteristics of Pratylenchus scribneri and Hoplolaimus galeatus indicated that there were patches within soybean fields in which both survival and reproduction wexe enhanced in spite of apparent homogeneity of soil type and topography. Treatment with carbofuran reduced the patchiness (or increased the dispersion) for H. galeatus while it had the opposite effect for P. scribneri. P. scribneri was less highly dispersed in conventional tillage plots than in the zero tillage plots. Populations from quadrats contained entirely within the patches could be described by the normal distribution (in the case of P. scribneri) or by the Poisson distribution (in the case of H. galeatus), while populations from quadrats contained entirely outside the patches could be described by the Poisson distribution for both nematodes. None of the distributions tested (Poisson, normal, negative binomial, Neyman's) gave an adequate fit when populations from both inside and outside the patches were considered together. In all instances, log₁₀ and ln transformations reduced the goodness of fit of the data to all of the distributions tested. Even with logarithmic transformations, parametric statistics were not appropriate for analysis of data in most instances.
nematode frequency; population ecology; index of dispersion; plot size selection; data transformation
Seashore paspalum (Paspalum vaginatum) has great potential for use in salt-affected turfgrass sites. Use of this grass on golf courses, athletic fields, and lawns in subtropical coastal areas may aid in conservation of freshwater resources. Belonolaimus longicaudatus and Hoplolaimus galeatus are considered among the most damaging root pathogens of turfgrasses in Florida. Glasshouse experiments were performed in 2002 and 2003 to examine the effects of increasing levels of irrigation salinity on B. longicaudatus and H. galeatus. Irrigation treatments were formulated by concentrating deionized water to six salinity levels (0, 5, 10, 15, 20, and 25 dS/m). Final population densities of H. galeatus followed a negative linear regression (r² = 0.92 and 0.83; P <= 0.01) with increasing salinity levels. Final population densities of B. longicaudatus were quadratically (r² = 0.72 and 0.78; P <= 0.01) related to increasing salinity levels from 0 to 25 dS/m. An increase in population densities of B. longicaudatus was observed at moderate salinity levels (10 and 15 dS/m) compared to 0 dS/m. Root-length comparisons revealed that B. longicaudatus caused root stunting at low salinity levels, 0 to 10 dS/m, but roots were not affected at 15 to 25 dS/m. These results indicate that the ability of B. longicaudatus to feed and stunt root growth was negatively affected at salinity levels of 15 dS/m and above.
Belonolaimus longicaudatus; Hoplolaimus galeatus; lance nematode; Paspalum vaginatum; salinity; seashore paspalum; sting nematode
The phylogenetic position of the human pathogenic fungus Penicillium marneffei was assessed from the nucleotide sequences of the nuclear and mitochondrial ribosomal DNA regions. Phylogenetic analysis determined that P. marneffei is closely related to species of Penicillium subgenus Biverticillium and sexual Talaromyces species with asexual biverticillate Penicillium states. Knowledge of the phylogenetic position of P. marneffei facilitated the design of unique oligonucleotide primers, from the nuclear ribosomal DNA internal transcribed spacer region, for the specific amplification of P. marneffei DNA. These primers were successful at selectively amplifying DNA from six isolates of P. marneffei and excluding the other species tested, which included Penicillium subgenus Biverticillium and Talaromyces species and several well-known fungal pathogens, namely, Aspergillus fumigatus, Coccidioides immitis, Histoplasma capsulatum, and Pneumocystis carinii. The primers that we have developed for the specific amplification of P. marneffei have the potential to be incorporated in a PCR identification system which could be used for the identification of this pathogenic agent from clinical material.
Hoplolaimus magnistylus n. sp. is described and illustrated. It was found in soil around roots of soybean in Arkansas and Mississippi. It is similar to H. galeatus and H. concaudajuvencus. It differs from H. galeatus in all stages primarily by possession of a longer stylet. It differs from H. concaudajuvencus by the possession of rounded tails in second-stage juveniles vs. conically pointed tails with acute termini, having fewer subdivisions in female basal lip annules, and the greater distance from female anterior end to posterior end of esophageal lobes. Morphometrics and descriptions of second-, third-, and fourth-stage juveniles are given. A paratype female of H. sheri was examined and found to have six esophageal gland nuclei.
taxonomy; morphology; new species
Three high-pressure liquid injection machines were used to inject isazofos into the root zone of tuff`grass plots to evaluate its potential for control of Hoplolaimus galeatus and Tylenchorhynchus dubius. A Rogers root zone injector delivering isazofos at 2.3 kg a.i./ha through 30° and 60° spray tips at 5,000 psi (3.45 × 10⁷ Pascals) significantly reduced nematode populations at 32 days after a single application and 33 days after a second application. In a second experiment with the Rogers injector at 2.3 kg a.i./ha, H. galeatus populations were significantly lower at 16 days after a single application and at 42 and 61 days after a second application with the 60° spray angle tips. An Envirojet turfgrass injector used to inject isazofos at 1.15 kg a.i./ha and 2.88 kg a.i./ha at 3,000 psi (1.38 × 107 Pascals) significantly reduced nematode populations at 7 days after treatment at the low rate and at 63 days after treatment with both application rates. A Landpride material injector applying isazofos at 6.9 and 13.8 kg a.i./ha at 2,000 psi (1.38 × 10⁷ Pascals) significantly reduced nematode populations at 7, 14, and 63 days after treatment at the high rate and at 63 days after the low-rate application. Although suppression of nematodes with isazofos was found, the degree of suppression is probably not enough to warrant recommendation of high-pressure delivery of isazofos for control of H. galeatus and T. dubius populations infesting turfgrasses.
Agrostis canina; Agrostis palustris; annual bluegrass; control; creeping bentgrass; high-pressure liquid injection; Hoplolaimus galeatus; isazofos; Lolium perenne; nematicide; nematode; perennial ryegrass; Poa annua; tuff`grass; Tylenchorhynchus dubius; velvet bentgrass
An identification key to 29 valid species of Hoplolaimus is given. A compendium of the most important diagnostic characters for use in identification of species is included as a practical alternative and supplement to the key. Diagnosis of Hoplolaimus is emended and lists of species of the genus, their synonymies, species inquirendae, nomina nuda, and species transferred to other genera are given. Hoplolaimus sheri, H. chambus, H. casparus, and H. capensis are recognized as valid species.
Compendium; diagnostic; Hoplolaimidae; Hoplolaimus; identification; key; lance nematode; morphology; Nematoda; taxonomy
Background and Aims
The genus Sinojackia consists of eight species, all endemic to China. All species of Sinojackia are endangered or threatened owing to poor recruitment within populations. Information on molecular phylogenetics is critical for developing successful conservation strategies for this genus.
Combined DNA sequence data from the nuclear ribosomal internal transcribed spacer regions and plastid psbA–trnH intergenic spacer and microsatellite data were used to infer a phylogeny of the genus.
Parsimony analysis of the combined sequence data and multivariate analysis based on fruit characters indicated that Sinojackia dolichocarpa is monophyletic and genetically well separated from the other Sinojackia species, thus supporting its rank at the generic level as Changiostyrax. Phylogenetic relationships within Sinojackia sensu stricto are unresolved from the combined sequence data. A UPGMA dendrogram based on seven microsatellite loci of 96 individual plants yielded a first-diverging cluster of all individuals of S. microcarpa. The remaining species form another cluster without any definitive patterns corresponding to current species circumscriptions, suggesting either extensive hybridization or incipient speciation.
The results suggest that there are too many species recognized within Sinojackia sensu stricto, but this must be further assessed with comprehensive morphological and taxonomic revisionary work. The implications of the phylogenetic data for conservation are discussed.
Changiostyrax; conservation; phylogeny; Sinojackia; Styracaceae
One hundred and eleven golf courses from 39 counties in the Carolinas were surveyed for plant-parasitic nematodes. Species diversity within habitats was analyzed with five diversity indices including Diversity index (H’), Evenness (J’), Richness (SR), Dominance (λ) and Diversity (H2). The results revealed a remarkably high diversity of 24 nematode species belonging to 19 genera and 11 families. Of those, 23 species were found in SC, 19 species in NC, and 18 species were detected in both states. Helicotylenchus dihystera, Mesocriconema xenoplax, Hoplolaimus galeatus, Tylenchorhynchus claytoni, Belonolaimus longicaudatus, Meloidogyne graminis and Paratrichodorus minor were the most prevalent and abundant species in golf course turfgrasses in both states. Twelve species were new records of plant parasitic nematodes in turfgrasses in both NC and SC. The results also revealed effects of different habitats on diversity of nematode species in turfgrass ecosystem. H’ and SR values were higher in SC than in NC. H’, J’ and H2 values were significantly higher in sandy than in clay soil in NC, but no significant differences between sand and clay soil were detected in SC or in pooled data from both states. There were no significant differences for all indices among the management zones (putting green, fairway and tee) in NC. However, in SC and pooled data, H’, SR and H2 were significantly higher in putting greens than in fairways and tees. Significant differences from different grass species (bermudagrass, creeping bentgrass and zoysiagrass) were detected only in H’, which was significantly higher in zoysiagrass than in bentgrass or bermudagrass in NC. In pooled data, H’ was significantly higher in zoysiagrass samples than in creeping bentgrass samples but was not significantly different from bermudagrass samples.
North Carolina; South Carolina; detection; distribution; diversity; ecology; golf course; identification; plant-parasitic nematode; turfgrass
Anopheles subpictus sensu lato is a major malaria vector in South and Southeast Asia. Based initially on polytene chromosome inversion polymorphism, and subsequently on morphological characterization, four sibling species A-D were reported from India. The present study uses molecular methods to further characterize and identify sibling species in Sri Lanka.
Mosquitoes from Sri Lanka were morphologically identified to species and sequenced for the ribosomal internal transcribed spacer-2 (ITS2) and the mitochondrial cytochrome c oxidase subunit-I (COI) genes. These sequences, together with others from GenBank, were used to construct phylogenetic trees and parsimony haplotype networks and to test for genetic population structure.
Both ITS2 and COI sequences revealed two divergent clades indicating that the Subpictus complex in Sri Lanka is composed of two genetically distinct species that correspond to species A and species B from India. Phylogenetic analysis showed that species A and species B do not form a monophyletic clade but instead share genetic similarity with Anopheles vagus and Anopheles sundaicus s.l., respectively. An allele specific identification method based on ITS2 variation was developed for the reliable identification of species A and B in Sri Lanka.
Further multidisciplinary studies are needed to establish the species status of all chromosomal forms in the Subpictus complex. This study emphasizes the difficulties in using morphological characters for species identification in An. subpictus s.l. in Sri Lanka and demonstrates the utility of an allele specific identification method that can be used to characterize the differential bio-ecological traits of species A and B in Sri Lanka.
Anopheles subpictus; Anopheles sundaicus; Cytochrome c oxidase subunit-I; ITS2; Malaria; Sibling species; Sri Lanka
Pinus edulis and Juniperus monosperma seedlings were inoculated separately with each of seven nematode species, and grown for 9 months at 20 C soil temperature. Hoplolaimus galeatus, Rotylenchus pumilis, Tylenchus exiguus, and Xiphinema americanum parasitized P. edulis seedlings, but did not significantly reduce seedling growth. Pinus edulis was not a host for Tylenchorhynchus cylindricus, Aphelenchoides cibolensis, or Criconemoides humilis. Xiphinema americanum and R. pumilis parasitized J. monosperma seedlings, and reduced their root weights and root collar diameters. Juniperus monosperma was not a host for A. cibolensis and T. exiguus, and parasitism of this tree species by T. cylindricus and C. humilis remains uncertain.
forest trees; tree seedlings; pine; juniper
Attachment of relatively low numbers of endospores from two isolates of Pasteuria spp. to several species of nematodes was consistently achieved in 2-5 minutes with a centrifugation technique. The rate of attachment of Pasteuria penetrans at 10⁴ endospores/0.1 ml/tube to second-stage juveniles (J2) of Meloidogyne javanica, M. incognita race 1, M. incognita race 3, and M. arenaria races 1 and 2 in two tests averaged 4.4, 5.2, 0.1, 0.3, and 0 endospores per J2, respectively. The rate of attachment Pasteuria sp. at 10³ endospores/0.1 ml/tube to individuals of Hoplolaimus galeatus, Belonolaimus longicaudatus, M. arenaria race 1, M. javanica, and M. incognita race 1 in two tests averaged 0.8, 0.04, 0, 0, and 0 endospores per nematode, respectively. The rate of attachment of P. penetrans to M. javanica at 10³, 10⁴, or 10⁵ endospores/0.1 ml/tube from two tests averaged 1.0, 5.7, and 28.3 endospores per J2, respectively. All of the J2 had endospores attached following centrifugation in tubes with 10⁴ and 10⁵ endospores/0.1 ml/tube.
bacterium; biological control; centrifugation; endospore; Meloidogyne arenaria; M. incognita; M. javanica; method; nematode; Pasteuria penetrans; Pasteuria sp.