The potato rot nematode (Ditylenchus destructor) is a very economically important nematode in agronomic and horticultural plants worldwide.  In this study, 43 populations of D. destructor were collected from different hosts across China, including 37 populations from Chinese herbal medicine plants.  Obtained sequences of ITS-rDNA and D2–D3 of 28S-rDNA genes of D. destructor were compared and analyzed.  Nine types of significant length variations in ITS sequences were observed among all populations.  The differences in ITS1 length were mainly caused by the presence of repetitive elements with substantial base substitutions.  Reconstructions of ITS1 secondary structures showed that the minisatellites formed a stem structure.  Ten haplotypes were observed in all populations based on mutations and variations of helix H9.  Among them, 3 known haplotypes (A–C) were found in 7 populations isolated from potato, sweet potato, and Codonopsis pilosula, and 7 unique haplotypes were found in other 36 populations collected from C. pilosula and Angelica sinensis compared with 7 haplotypes (A–G) according to Subbotin’ system.  These unique haplotypes were different from haplotypes A–G, and we named them as haplotypes H–N.  The present results showed that a total of 14 haplotypes (A–N) of ITS-rDNA have been found in D. destructor.  Phylogenetic analyses of ITS-rDNA and D2–D3 showed that all populations of D. destructor were clustered into two major clades: one clade only containing haplotype A from sweet potato and the other containing haplotypes B–N from other plants.  For further verification, PCR-ITS-RFLP profiles were conducted on 7 new haplotypes.  Collectively, our study suggests that D. destructor populations on Chinese medicinal materials are very different from those on other hosts and this work provides a paradigm for relevant researches.

The cereal cyst nematode, Heterodera avenae, is one of the most economically important pathogens impacting the worldwide production of cereals and is widely distributed in more than 16 regions in China.  The present study used the numbers of nematodes inside the plant roots to evaluate the resistance/susceptibility of different subpopulations of barley Hordeum vulgare (QH2R, QH6R and TB2R) to H. avenae under field and pot conditions.  Nematode development in two highly resistant varieties was also evaluated by in vivo experiment and microscopic observation.  Analyses of 186 selected varieties showed the numbers of susceptible varieties identified with the number of females/cysts per plant (NFP) method were significantly higher than those identified with the Pf/Pi ratio (PPR) method, which indicated that the NFP method rather than the PPR method is more reliable to evaluate the resistance of barley.  The field and pot experiment results indicated that the QH2R subpopulation had lower females/cysts numbers than QH6R and TB2R subpopulations, and eight HR varieties (Sunong 7617, Sunong 7635, Dongyuan 87-14, Rudong 14-46, Rudong 87-57, Rudong 87-8-45, Rudong 88-14-2, and Rudong 88-67-1) were identified in QH2R, with the NFP numbers below 4.2.  Further microscopic observation of nematode development suggested that H. avenae often penetrated less into highly resistant varieties (Sunong 7635 and Dongyuan 87-14) and more frequently failed to develop into females than the susceptible barleys.  The promising resistant varieties identified in the present research might be helpful for breeders to develop CCN-resistant cultivars and control H. avenae populations effectively at low costs.

Stunt nematodes (Tylenchorhynchus spp.) are obligate migratory root ecto-parasitic nematodes found in the fields of many cultivated crops.  These nematodes, with phyto-sanitary potential, are frequently ignored or misdiagnosed as pests, and this may pose a threat to food security.  The accuracy of its identification based on a morphological approach has been challenged recently, due to the overlapping of the morphological and morphometric characters of the species.  Consequently, the objective of this study is to identify and characterize stunt nematodes present in 54 fields cultivated with cereal crops (wheat, maize and rice) in the savannahs of northern Nigeria, using integrative taxonomy and molecular approaches.  The molecular and morphological studies identified and confirmed the presence of T. annulatus as the occurring specie in the savannahs of northern Nigeria.  The phylogenetic analysis was carried out using the internal transcribed spacer (ITS) and 28S genes of ribosomal DNA further confirmed the presence of T. annulatus.  The first molecular characterization and sequences of the ITS and 28S rDNA gene for T. annulatus from Nigeria were provided by this research.  Also, according to our literature search, this is the first report on T. annulatus in wheat, maize and rice in the savannahs of northern Nigeria.  Further study to test the pathogenicity of the parasitic nematode species found in this survey is recommended for the prioritization and development of efficient management strategies.

Potassium (K), an important nutrient element, can improve the stress resistance/tolerance of crops.  The application of K in resisting plant-parasitic nematodes shows that the K treatment can reduce the occurrence of nematode diseases and increase crop yield.  However, data on K₂SO₄ induced rice resistance against the root-knot nematode Meloidogyne graminicola are still lacking.  In this work, K₂SO₄ treatment reduced galls and nematodes in rice plants and delayed the development of nematodes.  Rather than affecting the attractiveness of roots to nematodes and the morphological phenotype of giant cells at feeding sites, such an effect is achieved by rapidly priming hydrogen peroxide (H₂O₂) accumulation and increasing callose deposition.  Meanwhile, galls and nematodes in rice roots were more in the potassium channel OsAKT1 and transporter OsHAK5 gene-deficient plants than in wild-type, while the K₂SO₄-induced resistance showed weaker in the defective plants.  In addition, during the process of nematode infection, the expression of jasmonic acid (JA)/ethylene (ET)/brassinolide (BR) signaling pathway-related genes and pathogenesis-related (PR) genes OsPR1a/OsPR1b was up-regulated in rice after K₂SO₄ treatment.  In conclusion, K₂SO₄ induced rice resistance against M. graminicola.  The mechanism of inducing resistance was to prime the basal defense and required the participation of the K⁺ channel and transporter in rice.  These laid a foundation for further study on the mechanism of rice defense against nematodes and the rational use of potassium fertilizer on improving rice resistance against nematodes in the field.

Soybean cyst nematode (SCN) Heterodera glycines is considered as the major constraint to soybean production.  GmSHMT08 at Rhg4 locus on chromosome 08, encoding a serine hydroxylmethyltransferase, is a major gene underlying resistance against H. glycines in Peking-type soybeans.  However, the molecular mechanism underpinning this resistance is less well characterized, and whether GmSHMT08 could interact with proteins in H. glycines remains unclear.  In this study, yeast two-hybrid screening was conducted using GmSHMT08 as a bait protein, and a fragment of a 70-kDa heat shock protein (HgHSP70) was screened from H. glycines that exhibited interaction with GmSHMT08.  This interaction was verified by both GST pull-down and bimolecular fluorescence complementation assays.  Our finding reveals HgHSP70 could be applied as a potential candidate gene for further exploring the mechanism on GmSHMT08-mediated resistance against SCN H. glycines.

The root-knot nematode Meloidogyne graminicola, which is distributed worldwide, is considered a major constraint on rice production in Asia.  The present study used the root gall index and number of nematodes inside the roots to evaluate resistance/susceptibility to M. graminicola in different subpopulations of Oryza sativa (aus, hybrid aus, indica, hybrid indica, temperate japonica, tropical japonica).  Nematode development in highly resistant varieties was also evaluated.  Analyses of randomly selected 35 varieties showed the number of M. graminicola nematodes inside the roots correlated very strongly (r=0.87, P≤0.05) with the nematode gall index, and the results from pot and field experiments revealed similar rankings of the varieties for resistance/susceptibility.  Among the 136 tested varieties, temperate japonica displayed the highest gall index, followed by tropical japonica, indica, hybrid indica, aus, and hybrid aus. Zhonghua 11 (aus), Shenliangyou 1 (hybrid aus) and Cliangyou 4418 (hybrid indica) were highly resistant to M. graminicola under both pot and field conditions.  Further examination of nematode development suggested that compared to susceptible rice, M. graminicola penetrated less often into highly resistant varieties and more frequently failed to develop into females.  The promising varieties found in the present research might be useful for the breeding of hybrid rice in China and for the further development of practical nematode management measures.   

Plant-parasitic nematodes are very common on cereal crops and cause economic losses via reduction in grain quality and quantity. During 2014, 83 soil samples were collected from wheat and barley fields in 21 districts of 13 provinces across five regions (Central Anatolia, Marmara, Aegean, Southeast Anatolia, and Black Sea Region) of Turkey. Cyst-forming nematodes were found in 66 samples (80%), and the internal transcribed spacer (ITS) sequencing and species-specific PCR identified the species in 64 samples as Heterodera filipjevi, Heterodera latipons, and Heterodera avenae. The predominant pathogenic cereal cyst nematode was H. filipjevi, which was found in all five regions surveyed. H. avenae was only detected in Southeast Anatolia whereas H. latipons was detected in Southeast Anatolia and Central Anatolia. ITS-rDNA phylogenetic analyses showed that H. avenae isolates from China clustered with H. australis, and Turkish isolates were closely related to European and USA isolates of this species. H. filipjevi from Turkey and China were clustered closely with those from the UK, Germany, Russia, and the USA. The density of many of these populations exceeded or approached the maximum threshold level for economic loss. To our knowledge, this is the first report of H. filipjevi in Diyarbakir, Edirne, and Kutahya provinces, and the first report of H. avenae in Diyarbakir Province. These results exhibit the most rigorous analysis to date on the occurrence and distribution of Heterodera spp. in Turkey’s major wheat-producing areas, thus providing a basis for more specific resistance breeding, as well as other management practices.

The cereal cyst nematode (CCN, Heteroder aavenae) causes serious yield loss on cereal crops, especially wheat, worldwide. Daxing population in Beijing City and Huangyuan population in Qinghai Province, China, are two CCN populations. In this study, the CCN pathotypes of Daxing and Huangyuan populations were characterized by tests on 23 standard “International Test Assortment” with the local species Wenmai 19 as the susceptible control. Tested materials were grouped by three nematode populations’ virulence on resistant genes (Rha1, Rha2, Rha3, Cre1) and nonresistant genes, varieties and lines. Both Daxing and Huangyuan populations were avirulent to Ortolan (Ha1). Barley cvs. Ortolan, Siri, Morocco, Bajo Aragon 1-1, and Martin 403-2 were all resistant to both populations. Cultivars Herta, Harlan 43 and wheat Iskamish-K-2-light were all susceptible to Huangyuan population, all of them, however, were resistant to Daxing population. The other five oats were all resistant to the two tested CCN populations. Except Iskamisch K-2-light, all the other wheat cultivars (Capa, Loros×Koga, AUS 10894, and Psathias) were susceptible to Daxing population. Because the pathotypes of the two tested CCN populations in Beijing and Qinghai were not identical to any of the 13 pathotypes previously characterized by the test assortment, we classified Daxing and Huangyuan populations as the new pathotypes, named Ha91.