Virginia R. Ferris

Association of RFLP markers with loci conferring broad-based resistance to the soybean cyst nematode (Heterodera glycines)

Soybean cyst nematode (SCN) is a major soybean yield-limiting pest. The present study was conducted to map broad-based SCN resistance loci from the cultivar ‘Hartwig’. Two-hundred F2∶3 lines derived from the cross ‘Williams 82’ x ‘Hartwig’ were screened with a fourth-generation SCN inbred and 56 polymorphic molecular markers. Allele states and phenotypes were analyzed using stepwise regression and the model selection was made at P ⩽ 0.01. Four unlinked RFLP markers (A006, A567, A487, A112) were associated with SCN resistance and the partial coefficient of determinations (R2) were 91%, 1%, 1%, and 1%. We have mapped a new, major SCN resistance locus (A006) and three minor loci (A567, A487, A112). This complete mapping will accelerate the transfer of broad-based resistance without linkage drag and aid in the determination of relationships among various SCN-resistant germplasm sources.

MicroRNA Discovery and Analysis of Pinewood Nematode Bursaphelenchus xylophilus by Deep Sequencing

BACKGROUND MicroRNAs (miRNAs) are considered to be very important in regulating the growth, development, behavior and stress response in animals and plants in post-transcriptional gene regulation. Pinewood nematode, Bursaphelenchus xylophilus, is an important invasive plant parasitic nematode in Asia. To have a comprehensive knowledge about miRNAs of the nematode is necessary for further in-depth study on roles of miRNAs in the ecological adaptation of the invasive species. METHODS AND FINDINGS Five small RNA libraries were constructed and sequenced by Illumina/Solexa deep-sequencing technology. A total of 810 miRNA candidates (49 conserved and 761 novel) were predicted by a computational pipeline, of which 57 miRNAs (20 conserved and 37 novel) encoded by 53 miRNA precursors were identified by experimental methods. Ten novel miRNAs were considered to be species-specific miRNAs of B. xylophilus. Comparison of expression profiles of miRNAs in the five small RNA libraries showed that many miRNAs exhibited obviously different expression levels in the third-stage dispersal juvenile and at a cold-stressed status. Most of the miRNAs exhibited obviously down-regulated expression in the dispersal stage. But differences among the three geographic libraries were not prominent. A total of 979 genes were predicted to be targets of these authentic miRNAs. Among them, seven heat shock protein genes were targeted by 14 miRNAs, and six FMRFamide-like neuropeptides genes were targeted by 17 miRNAs. A real-time quantitative polymerase chain reaction was used to quantify the mRNA expression levels of target genes. CONCLUSIONS Basing on the fact that a negative correlation existed between the expression profiles of miRNAs and the mRNA expression profiles of their target genes (hsp, flp) by comparing those of the nematodes at a cold stressed status and a normal status, we suggested that miRNAs might participate in ecological adaptation and behavior regulation of the nematode. This is the first description of miRNAs in plant parasitic nematodes. The results provide a useful resource for further in-depth study on molecular regulation and evolution of miRNAs in plant parasitic nematodes.

First Report of Soybean Cyst Nematode Reproduction on Purple Deadnettle under Field Conditions

Soybean cyst nematode commonly infests soybean fields in the United States and is a threat to profitable soybean production. Purple deadnettle is common in Indiana production fields and frequently occurs at very high densities. Purple deadnettle plants were removed from a research site infested with SCN. Peadnettle roots were found to possess cysts containing eggs. The morphology of cysts and juveniles was consistent with SCN. Based on the distribution of other cyst nematodes in Indiana, a bioassay on soybean from the field, and morphometric observations, we determined the identity of the nematode to be SCN.

Comparative transcriptomics of two pathogenic pinewood nematodes yields insights into parasitic adaptation to life on pine hosts

Bursaphelenchus xylophilus and Bursaphelenchus mucronatus are migratory endoparasitic nematodes that live in pine trees. To gain insight into their molecular similarities and differences, transcriptomes of the two nematodes were analysed. A total of 23,765 and 21,782 contigs (>300 bp) were obtained from B. xylophilus and B. mucronatus, respectively. More than 80% of the contigs could map to each others transcriptome reciprocally. A total of 23,467 and 21,370 Open Reading Frames were predicted, respectively. Besides those known parasitism-related proteins, six new venom allergen-like proteins (VAPs) were found, which were not homologous to known VAPs. Enzymes involved in xenobiotic biodegradation were abundant in the two transcriptomes based on KEGG functional annotation. Metabolism of xenobiotics by cytochrome P450 comprised the main detoxification pathways. The mRNA expression levels of detoxification genes in nematodes living in the host were higher than those in nematodes feeding on fungus. However, there were fewer enzymes involved in the α-pinene degradation. Our results indicate that the two pinewood nematodes have evolved similar molecular mechanisms to adapt to life on pine hosts.

Influence of Winter Annual Weed Management and Crop Rotation on Soybean Cyst Nematode (Heterodera Glycines) and Winter Annual Weeds

Abstract Certain winter annual weeds have been documented as alternative hosts to soybean cyst nematode (SCN), and infestations of such species have become common in no-till production fields in the Midwest. This research was conducted to determine the influence of herbicide- and cover-crop-based winter annual weed management systems and crop rotation on winter annual weed growth and seed production, SCN population density, and crop yield. Two crop rotations (continuous soybean and soybean-corn) and six winter annual weed management systems (a nontreated control, fall and spring herbicide applications, spring-applied herbicide, fall-applied herbicide, fall-seeded annual ryegrass, and fall-seeded winter wheat) were evaluated in no-tillage systems from fall 2003 to 2006 at West Lafayette, IN and Vincennes, IN. Fall or spring herbicide treatments generally resulted in lower winter annual weed densities than cover crops. Densities of henbit and purple deadnettle increased over years in the cover crop systems but remained constant in the herbicide systems. Averaged over sites and years, winter annual weed densities were nearly 45% lower in the spring than the fall due to winter mortality. Corn yield was reduced by the cover crops at West Lafayette but not Vincennes. Winter annual weed management system had no influence on soybean yield. SCN population density was reduced by including corn in the crop sequence but was not influenced by winter annual weed management. The density of weedy host species of SCN in the experimental area was relatively low (less than 75 plants m−2) compared to densities that can be observed in production fields. The results of these experiments suggest that inclusion of corn into a cropping sequence is a much more valuable SCN management tool than winter annual weed management. In addition, control of winter annual weeds, specifically for SCN management, may not be warranted in fields with low weed density. Nomenclature: Soybean cyst nematode, Heterodera glycines Ichinohe; corn, Zea mays L.; soybean, Glycine max (L.) Merr; wheat, Triticum aestivum L.

A new species and new synonym in Heptagenia walsh (Ephemeroptera: Heptageniidae: Heptageniinae) based on molecular and morphological evidence.

Abstract A new mayfly species, Heptagenia whitingi Webb & McCafferty n.sp. is described from larvae, a male subimago, a female adult, and eggs collected from large rivers in the west-central portion of North America. Larvae are differentiated from other North American Heptagenia Walsh by a pair of large, rectangular pale markings on abdominal tergum 4, and the combination of having the posterior margin of the abdominal terga with bluntly pointed spines less than half the length of the fine setae, small blunt spines on the posterior margin of the caudal filaments, and numerous rows of setae laterally on the ventral surface of the labrum. A 630 bp partial sequence of the mitochondrial gene cytochrome oxidase 1 (COI) from three specimens of H. whitingi n.sp. was compared with those of 12 specimens representing eight other North American species of Heptagenia. Intraspecific sequence divergences based on Kimura-2-parameter (K2P) distance ranged from 0–1.1%. Interspecific sequence divergence based on K2P distance ranged from 8.9–20.0%. Heptagenia whitingi n.sp. differed from its sister taxon H. flavescens (Walsh) by 11.7%. Heptagenia diabasia Burks and H. elegantula (Eaton) differed from each other by only 1.1%; these two alleged species show a clinal pattern in larval abdominal coloration and there are no structural differences between the semaphorants. On this basis, H. diabasia is placed as a junior subjective synonym of H. elegantula, n.syn.

Development of Soybean Cyst Nematode on Henbit (Lamium amplexicaule) and Purple Deadnettle (Lamium Purpureum)

A survey of seven production fields in Indiana, Illinois, and Ohio was conducted to assess henbit and purple deadnettle growth and soybean cyst nematode (SCN) development and reproduction on these weeds. Autumn and spring growth of purple deadnettle and henbit was influenced by location within each state. In general, winter annual weeds were larger in size and reached maturity earlier in the spring at the southern sample sites than those in the north. All growth stages of SCN were found to be associated with henbit and purple deadnettle at both autumn and spring sample timings. SCN juveniles were generally found infecting roots at highest abundance in the spring. SCN cyst and egg production also were widespread and occurred to a much higher degree during the autumn than the spring developmental period. The results of this survey indicate that management tactics designed to minimize the potential for SCN reproduction on winter annual weeds would probably be most effective if conducted in the autumn, when the majority of SCN reproduction occurred. However, spring populations of winter annual weeds that harbor SCN juveniles might facilitate additional SCN reproduction and population increase if the weeds are not controlled in a timely manner prior to planting. Nomenclature: Henbit, Lamium amplexicaule L. LAMAM; purple deadnettle, Lamium purpureum L. LAMPU; soybean, Glycine max (L.) Merr; soybean cyst nematode, Heterodera glycines Ichinohe.

Biometric Analyses in the Genus Labronema (Nematoda: Dorylaimida) With a Description of L. Thornei N. Sp. 1)

Biometrics for Labronema ferox Thorne 1939 and L. thornei n. sp., reared on agar substrates are given. L. ferox completes its life cycle under laboratory conditions in 3-4 months. L. thornei takes 5-6 months to complete its life cycle under similar conditions. Both species exhibit a pronounced heteromorphism in shape and length of tail. L. thornei, a monosexual species, is described.

Tests of current hypotheses of mayfly (Ephemeroptera) phylogeny using molecular (18s rDNA) data

Abstract Partial 18s rDNA sequences from 22 exemplar mayfly species (Ephemeroptera) representing 20 families were analyzed to obtain a best phylogenetic tree for comparison to previous phylogenetic hypotheses. With respect to relationships among the three major groupings, our molecular data support the hypothesis that Pisciforma and Setisura comprise a monophyletic sister group to the Furcatergalia, rather than the hypothesis that Setisura and Furcatergalia comprise a monophyletic group stemming from the Pisciforma. Within Pisciforma, acceptable trees show that Baetidae separates at the base of the Pisciforma clade. The data suggest that Pisciforma is paraphyletic and do not support the grouping of all Southern hemisphere families as a monophyletic group. An evolutionary sequence favored by the data suggests a grouping of Siphlonuridae, Rallidentidae, Nesameletidae, and Ameletidae and a grouping of Oniscigastridae, Ameletopsidae, and Acanthametropodidae. The data support the monophyly of Setisura (Heptageniidae, Arthropleidae, Pseudironidae, Oligoneuriidae, Isonychiidae, and Coloburiscidae). Within Setisura, a bootstrap/jackknife test places the families Heptageniidae, Arthropleidae, and Pseudironidae in one clade at 100% frequency. Also supported are hypotheses that Pseudironidae is a sister group to a Heptageniidae–Arthropleidae group and that a sister relationship exists between the latter two families. Hypotheses that Pseudironidae separated from other Setisura families at an earlier stage and comprises a sister group to a Heptageniidae–Oligonuriidae lineage or that Pseudironidae should be moved out of Setisura are not supported.

Influence of Intraspecific Henbit (Lamium amplexicaule) and Purple Deadnettle (Lamium purpureum) Competition on Soybean Cyst Nematode Reproduction

Abstract A greenhouse study was conducted to determine the effect of henbit and purple deadnettle density on weed biomass accumulation and soybean cyst nematode (SCN) reproduction. SCN did not impact shoot or root dry weight of purple deadnettle, henbit, or soybean. Foliar and root biomass of henbit and purple deadnettle were comparable but the biomass per stem was higher for purple deadnettle. Shoot and root biomass per pot of henbit and purple deadnettle at corresponding plant densities were statistically similar and were generally higher with increasing plant density. Henbit produced a greater number of stems than purple deadnettle and the least number of stems for both species existed at low densities. Purple deadnettle allowed for more SCN reproduction than did henbit. Weed densities also influenced SCN cyst and egg production but the results were species dependent. The highest SCN reproduction per pot was supported at low to moderate densities of purple deadnettle but at moderate to high densities of henbit. These results suggest that purple deadnettle should be more aggressively managed than henbit in management programs for SCN, but that henbit, especially at high densities, can support SCN reproduction at levels near those of purple deadnettle. Nomenclature: Henbit, Lamium amplexicaule L. LAMAM; purple deadnettle Lamium purpureum L. LAMPU; soybean Glycine max (L.) Merr. ‘Williams 82’ and ‘PI437654’