Classification Methods and Identification of Reniform Nematode Resistance in Known Soybean Cyst Nematode-Resistant Soybean Genotypes.

Plant parasitic nematodes are a major yield-limiting factor of soybean in the United States and Canada. It has been indicated that soybean cyst nematode (SCN; Heterodera glycines Ichinohe) and reniform nematode (RN; Rotylenchulus reniformis Linford and Oliveira) resistance could be genetically related. For many years, fragmentary data have shown this relationship. This report evaluates RN reproduction on 418 plant introductions (PIs) selected from the U.S. Department of Agriculture Soybean Germplasm Collection with reported SCN resistance. The germplasm was divided into two tests of 214 PIs reported as resistant and 204 PIs reported as moderately resistant to SCN. The defining and reporting of RN resistance changed several times in the last 30 years, causing inconsistencies in RN resistance classification among multiple experiments. Comparison of four RN resistance classification methods was performed: (i) ≤10% as compared with the susceptible check, (ii) using normalized reproduction index (RI) values, and using (iii) transformed data log10(x), and (iv) transformed data log10(x + 1) in an optimal univariate k-means clustering analysis. The method of transformed data log10(x) was selected as the most accurate for classification of RN resistance. Among 418 PIs with reported SCN resistance, the log10(x) method grouped 59 PIs (15%) as resistant and 130 PIs (31%) as moderately resistant to RN. Genotyping of a subset of the most resistant PIs to both nematode species revealed their strong correlation with rhg1-a allele. This research identified genotypes with resistance to two nematode species and potential new sources of RN resistance that could be valuable to breeders in developing resistant cultivars.

Dissecting nematode resistance regions in soybean revealed pleiotropic effect of soybean cyst and reniform nematode resistance genes.

Reniform nematode (RN, Rotylenchulus reniformis Linford & Oliveira) has emerged as one of the most important plant parasitic nematodes of soybean [Glycine max (L.) Merr.]. Planting resistant varieties is the most effective strategy for nematode management. The objective of this study was to identify quantitative trait loci (QTL) for RN resistance in an exotic soybean line, PI 438489B, using two linkage maps constructed from the Universal Soybean Linkage Panel (USLP 1.0) and next-generation whole-genome resequencing (WGRS) technology. Two QTL controlling RN resistance were identified-the soybean cyst nematode (SCN, Heterodera glycines) resistance gene GmSNAP18 at the rhg1 locus and its paralog GmSNAP11. Strong association between resistant phenotype and haplotypes of the GmSNAP11 and GmSNAP18 was observed. The results indicated that GmSNAP11 possibly could have epistatic effect on GmSNAP18, or vice versa, with the presence of a significant correlation in RN resistance of rhg1-a GmSNAP18 vs. rhg1-b GmSNAP18. Most importantly, our preliminary data suggested that GmSNAP18 and GmSNAP11 proteins physically interact in planta, suggesting that they belong to the same pathway for resistance. Unlike GmSNAP18, no indication of GmSNAP11 copy number variation was found. Moreover, gene-based single nucleotide polymorphism (SNP) markers were developed for rapid detection of RN or SCN resistance at these loci. Our analysis substantiates synergic interaction between GmSNAP11 and GmSNAP18 genes and confirms their roles in RN as well as SCN resistance. These results could contribute to a better understanding of evolution and subfunctionalization of genes conferring resistance to multiple nematode species and provide a framework for further investigations.

Plant elicitor peptides promote plant defences against nematodes in soybean.

Plant elicitor peptides (Peps) are widely distributed among angiosperms, and have been shown to amplify immune responses in multiple plant families. Here, we characterize three Peps from soybean (Glycine max) and describe their effects on plant defences against two damaging agricultural pests, the root-knot nematode (Meloidogyne incognita) and the soybean cyst nematode (Heterodera glycines). Seed treatments with exogenous GmPep1, GmPep2 or GmPep3 significantly reduced the reproduction of both nematodes. Pep treatment also protected plants from the inhibitory effects of root-knot nematodes on above-ground growth, and up-regulated basal expression levels of nematode-responsive defence genes. GmPep1 induced the expression of its propeptide precursor (GmPROPEP1), a nucleotide-binding site leucine-rich repeat protein (NBS-LRR), a pectin methylesterase inhibitor (PMEI), Respiratory Burst Oxidase Protein D (RBOHD) and the accumulation of reactive oxygen species (ROS) in leaves. In addition, GmPep2 and GmPep3 seed treatments up-regulated RBOHD expression and ROS accumulation in roots and leaves. These results suggest that GmPeps activate plant defences through systemic transcriptional reprogramming and ROS signalling, and that Pep seed treatments represent a potential strategy for nematode management.

Advancements in breeding, genetics, and genomics for resistance to three nematode species in soybean.

KEY MESSAGE: Integration of genetic analysis, molecular biology, and genomic approaches drastically enhanced our understanding of genetic control of nematode resistance and provided effective breeding strategies in soybeans. Three nematode species, including soybean cyst (SCN, Heterodera glycine), root-knot (RKN, Meloidogyne incognita), and reniform (RN, Rotylenchulus reniformis), are the most destructive pests and have spread to soybean growing areas worldwide. Host plant resistance has played an important role in their control. This review focuses on genetic, genomic studies, and breeding efforts over the past two decades to identify and improve host resistance to these three nematode species. Advancements in genetics, genomics, and bioinformatics have improved our understanding of the molecular and genetic mechanisms of nematode resistance and enabled researchers to generate large-scale genomic resources and marker-trait associations. Whole-genome resequencing, genotyping-by-sequencing, genome-wide association studies, and haplotype analyses have been employed to map and dissect genomic locations for nematode resistance. Recently, two major SCN-resistant loci, Rhg1 and Rhg4, were cloned and other novel resistance quantitative trait loci (QTL) have been discovered. Based on these discoveries, gene-specific DNA markers have been developed for both Rhg1 and Rhg4 loci, which were useful for marker-assisted selection. With RKN resistance QTL being mapped, candidate genes responsible for RKN resistance were identified, leading to the development of functional single nucleotide polymorphism markers. So far, three resistances QTL have been genetically mapped for RN resistance. With nematode species overcoming the host plant resistance, continuous efforts in the identification and deployment of new resistance genes are required to support the development of soybean cultivars with multiple and durable resistance to these pests.

Identification of quantitative trait loci underlying resistance to southern root-knot and reniform nematodes in soybean accession PI 567516C.

Soybean cyst nematode (SCN, Heterodera glycine Ichinohe), southern root-knot nematode [SRKN, Meloidogyne incognita (Kofoid and White) Chitwood] and reniform nematode (RN, Rotylenchulus reniformis Linford and Oliveira) are three important plant-parasitic pests in soybean. Previous study showed that plant introduction (PI) 567516C harbored novel quantitative trait loci (QTL) conferring SCN resistance to soybean. However, QTL underlying resistance to SRKN and RN in PI 567516C remain unknown. The objectives of this study were to identify QTL for resistance to SRKN and RN in PI 567516C. Two hundred and forty-seven F6:9 recombinant inbred lines, derived from a cross between cultivar Magellan and PI 567516C, were evaluated for resistance to SRKN and RN. Two hundred and thirty-eight simple sequence repeats and 687 single nucleotide polymorphism markers were used to construct a genetic linkage map. Three significant QTL associated with resistance to SRKN were mapped on chromosomes (Chrs.) 10, 13 and 17. Two significant QTL associated with resistance to RN were detected on Chrs. 11 and 18. Whole-genome resequencing revealed that there might be Peking-type Rhg1 in PI 567516C. Our study provides useful information to employ PI 567516C in soybean breeding in order to develop new cultivars with resistance to multiple nematodes.

Taxonomic and molecular identification of hemicaloosia, hemicycliophora, gracilacus and paratylenchus species (nematoda: criconematidae).

Populations of Hemicycliophora epicharoides, H. gigas, H. labiata, H. pruni, H. shepherdi, H. vidua, H. zuckermani, Gracilacus straeleni, and Paratylenchus labiosus were obtained from different geographical areas in the continental United States and characterized morphological and molecularly. Two new species of Hemicycliophorinae: Hemicaloosia uarki n. sp from Pinetree, St. Francis County, Arkansas, and Hemicycliophora wyei n. sp from Wayne County, North Carolina, are also described. Hemicaloosia uarki n. sp. is characterized by having two lip annuli separated from the rest of body and directed anteriorly, a long stylet (106-124 µm), long body length (1,081-1,326 µm) and a single lateral fields demarcated by interruptions of the body annuli. Hemicycliophora wyei n. sp. showed a lateral fields demarked by two faint lines with transverse anastomoses and/or breaks of the striae; an elongated not offset conical tail with distinct annulations and a rounded tip and long vulval lips with a vulval sleeve. The molecular characterizations of the new (H. uarki n. sp. and H. wyei n. sp.) and known species of Criconematidae using the ITS1 rDNA gene sequence and the molecular phylogenetic relationships are provided.

Taxonomic and molecular identification of bakernema, criconema, hemicriconemoides, ogma and xenocriconemella species (nematoda: criconematidae).

Populations of Bakernema inaequale, C. petasum, C. sphagni, C. mutabile, Ogma octangulare, Xenocriconemella macrodora and Hemicriconemoides chitwoodi were identified and re-described from different geographical areas in the continental United States and molecularly characterized. Two new species of spine nematodes Criconema arkaense n. sp. from Washington County and Lee County, Arkansas and Criconema warrenense n. sp from Warren, Bradley County, Arkansas are also described and named. Criconema arkaense is characterize by having a conspicuous lip region offset from the body with two annuli, short rounded tail with a thin cuticular sheath and subterminal anus. Criconema warrenense n. sp. has two lip region annuli about the same width, first annulus directed posteriorly, separated by a narrow neck annulus and a short conoid tail, unilobed non-folded annulus. The molecular characterization of Criconema arkaense and Criconema warrenense using ITS1 rDNA gene sequence and the molecular phylogenetic relationships of these new species along with the known spines nematodes are provided.

Taxonomic and molecular identification of mesocriconema and criconemoides species (nematoda: criconematidae).

Populations of Mesocriconema curvatum, M. kirjanovae, M. onoense, M. ornatum, M. sphaerocephala, M. surinamense, M. vadense, M. xenoplax, and Criconemoides informis from different geographical areas in the continental United States were characterized morphologically and molecularly. A new ring nematode from Washington County, Arkansas, is also described and named Mesocriconema ozarkiense n. sp., This new species is characterized by females with small flattened submedian lobes, lower than or at the same level as the labial disc, vagina straight, very well developed spermatheca without sperm, no more than one anastomoses, L=379-512 µm, V=89-93, stylet length = 49-61 µm, R=107-119, annuli with slightly crenate margins on tail portion and a simple anterior vulval lip. The molecular characterization of M. ozarkiense n. sp. using the ITS rRNA gene sequence and the phylogenesis relationship of this new species with the ring nematodes included in this study are provided.

Longidorus kheirii n. sp. (Nematoda: Longidoridae) from Iran.

Longidorus kheirii n. sp., a parthenogenetic species, was found in soil samples collected from the rhizosphere of Rosa sp. growing in a natural mountainous region close to Maragheh city, northwestern Iran. It is characterised by having a long body (6.7-9 mm), a 19.5-23 mum wide head continuous with the body contour, a truncate and slightly concave lip region with convex sides between the anterior end and the guide-ring, an odontostyle 113-130 mum long, an odontophore 69-97.5 mum long, a body width of 90.5-117.5 mum at the mid-body, a long, wide oesophageal bulb (149.5-193.5 x 39.5-48 mum), a tail length of 47-72 mum, a male with 11 ventromedian supplements and spicules of 85 mum in length, and four juvenile stages. The ribosomal 18S rDNA gene of L. kheirii n. sp., L. leptocephalus Hooper, 1961, L. profundorum Hooper, 1966 L. euonymus Mali & Hooper, 1973 and two unidentified species listed as Longidorus sp. 1 and Longidorus sp. 2, all recovered from northwestern Iran in the same survey, and the ITS1 of L. kheirii n. sp. and Longidorus sp. 1 were sequenced in order to investigate the phylogenetic relationships with other previously sequenced Longidorus species.

Intraspecific Variability of Rotylenchulus reniformis from Cotton-growing Regions in the United States.

Reniform nematode (Rotylenchulus reniformis) is a major pest of cotton in the southeastern United States. The objective of this study was to examine the variation of reniform nematode populations from cotton-growing locations in the United States where it is prevalent. Multivariate analysis of variance and discriminant analysis were used to determine the variability of morphology in males and immature females. Reproduction indices of populations were measured on selected soybean and cotton genotypes in the greenhouse. High variability in morphometrics and reproduction was observed within all the populations, and several differences were found among populations. DNA sequences of the nuclear ribosomal first internal transcribed spacer region (ITS1) were compared among populations from the United States and to sequences of populations from Brazil, Colombia, Honduras, and Japan. No polymorphic nucleotide sites were observed among the amphimictic populations. Only a parthenogenic population from Japan was distinct. The phenotypic polymorphism of the species in the United States could impact the effectiveness of management strategies based on host plant resistance.

Histological Changes in Gossypium hirsutum Associated with Reduced Reproduction of Rotylenchulus reniformis.

The reniform nematode (Rotylenchulus reniformis) is an important parasite of upland cotton (Gossypium hirsutum). Parasitism involves the formation of syncytia to provide nutrition for the female. Events that occur at the feeding site may determine the degree of susceptibility of cotton plants to reniform nematode. The objective of this work was to describe histological modifications associated with reduced reproduction of Rotylenchulus reniformis in upland cotton roots. 'Deltapine 50' cotton and a selection from this line with a moderate level of resistance were inoculated with reniform nematode in the greenhouse, and observations on roots were made 3, 6, 9, 12, and 15 days after inoculation. No differences in penetration behavior or in the formation and characteristics of syncytia were observed. Reduced reproduction was correlated with an earlier degeneration and collapse of the syncytial cells, and occasionally, with lack of hypertrophy of the pericycle cells involved. These two mechanisms accounted for 40% to 60% reduction of reproduction of reniform nematode in the plants examined.

Histological Observations of Rotylenchulus reniformis on Gossypium longicalyx and Interspecific Cotton Hybrids.

Observations on the development of reniform nematode (Rotylenchulus reniformis) on roots of Gossypium longicalyx, G. hirsutum, and two interspecific hybrids derived from them were made by light microscopy. Gossypium longicalyx is reported to be immune to reniform nematode, but the mechanism(s) for resistance are unknown. Penetration of G. longicalyx roots by female nematodes was confirmed, and incipient swelling of the females, indicating initiation of maturation of the reproductive system, was observed. Female maturation occurred up to the formation of a single embryo inside the female body but not beyond this point. In both hybrids, development was inhibited but progressed further than in the immune parent. Reactions ranged from highly compatible, with the formation of active syncytia and full development of females, to incompatible with little or no development of the female. Compatible plants showed characteristic hypertrophied cells, enlarged nuclei, dense cytoplasm, and partial dissolution of cell walls, whereas incompatible plant reactions included lignification of the cells adjacent to the nematode head, or the complete collapse and necrosis of the cells involved. The need to characterize reactions and to carefully select among the plants descended from the hybrids during the introgression process, as well as the importance of combining the results of reproduction tests with histological observation of the plant-nematode interactions, is discussed.