Root-Knot Nematodes and the Process of Ageing in Plants

Infection of plants by root-knot nematodes is often accompanied by physiological changes characteristic of ageing. Ultra-low tissue luminescence of infected plants indicated oxidation of cell-membrane lipids. Cells with membranes subjected to oxidation lose some of their capacity for water retention. Treating tomato and radish with lidocaine hydrochloride, an inhibitor of lipid oxidation, retarded above-ground symptoms of root-knot nematode infection and of ageing.     

Isolation and Characterization of a Rhizobacterial Antagonist of Root-Knot Nematodes

The rhizobacterial strain Jdm2 was isolated from the rhizosphere of the traditional Chinese medicinal herb Trichosanthes kirilowii in Jiangsu province, China, and was identified as Bacillus subtilis. Exposure of cell-free filtrate of the strain to the root-knot nematode Meloidogyne incognita under in vitro conditions caused substantial mortality of the second stage juvenile (J2) and significantly reduced egg hatchability. A greenhouse trial demonstrated that 56 days after treatment with Jdm2, the number of galls associated with M. incognita infection in the tomato (Solanum lycopersicum) roots was significantly reduced compared to controls, and the disease severity of infected plants was lower in treated plants (36%) compared to water control (75%). Consistently, in the field trial, the biocontrol efficacy of Jdm2 reached 69%, 51% and 48% after 30, 60 and 90 days post-transplantation, respectively. As indicated by PCR-DGGE analysis, inoculation with Jdm2 strain had an effect on the bacterial community of the tomato rhizosphere at the first stage, but was not able to imperil the bacterial community stability for long time. The novel bacterial strain Jdm2 enhances plant growth and inhibits nematode activity, and has the potential to be a safe and effective microbial pesticide.     

猕猴桃属种间嫁接亲和性试验研究及抗根结线虫砧木的初步筛选

以中华猕猴桃（Actinidia chinensis)武植2号（Wuzhi-2),通山5号（Tongshan-5)品种和美味猕猴桃（A.deliciosa)金魁（Jinkui)品种为接穗,采用美味猕猴桃等9种类型猕猴桃一年生实生苗为砧木进行嫁接,研究其短期嫁接亲和性,并对这9种砧木进行抗根结线虫（Meloidogyne)筛选试验。结果表明,种间的嫁接亲和性及抗根线虫性 均有显著差异：以美味猕猴桃和中华猕猴桃优良品种实生苗为砧木,与3个品种均有高的亲和性,成活率高达100％,且抗性在9种砧木中最强;其它物种作砧木均有不足,武植2号／山猴猴桃,通山5号／M16,通山5号／山梨猕猴桃3个嫁接组合的亲和性均较好,成活率在80％以上,但山梨猕猴桃较易感病且与金魁的亲和力较低,M16抗性虽较好,但与金魁,武植2号的亲和力也较低,漓江猕猴桃是抗性较强的物种,但它与3种接穗品种的亲和力都较小,其它5种砧木,对根结线虫的抗性均不强,因此结合成活率及抗根结线虫两因素,在供试材料中,美味猴猕桃和中华猕猴桃接适应性广且抗性强,是较好的砧木或选育砧木的亲本材料。     

Effect of Emamectin Benzoate on Root-Knot Nematodes and Tomato Yield

Southern root-knot nematode (Meloidogyne incognita) is an obligate, sedentary endoparasite of more than 3000 plant species, that causes heavy economic losses and limit the development of protected agriculture of China. As a biological pesticide, emamectin benzoate has effectively prevented lepidopteran pests; however, its efficacy to control M. incognita remains unknown. The purpose of the present study was to test soil application of emamectin benzoate for management of M. incognita in laboratory, greenhouse and field trials. Laboratory results showed that emamectin benzoate exhibited high toxicity to M. incognita, with LC₅₀ and LC₉₀ values 3.59 and 18.20 mg L^-1, respectively. In greenhouse tests, emamectin benzoate soil application offered good efficacy against M. incognita while maintaining excellent plant growth. In field trials, emamectin benzoate provided control efficacy against M. incognita and resulted in increased tomato yields. Compared with the untreated control, there was a 36.5% to 81.3% yield increase obtained from all treatments and the highest yield was received from the highest rate of emamectin benzoate. The results confirmed that emamectin benzoate has enormous potential for the control of M. incognita in tomato production in China.     

Lateral Transfers of Serine Hydroxymethyltransferase (glyA) and UDP-N-Acetylglucosamine Enolpyruvyl Transferase (murA) Genes from Free-living Actinobacteria to the Parasitic Chlamydiae

The chlamydiae are important human and animal pathogens which form a phylogentically distinct lineage within the Bacteria. There is evidence that some genes in these obligate intracellular parasites have undergone lateral exchange with other free-living organisms. In the present work, we describe two interesting cases of lateral gene transfer between chlamydiae and actinobacteria, which have been identified based on the shared presence of conserved inserts in two important proteins. In the enzyme serine hydroxymethyltransferase (SHMT or GlyA protein), which links amino acid and nucleotide metabolisms by generating the key intermediate for one-carbon transfer reactions, two conserved inserts of 3 and 31 amino acids (aa) are uniquely present in various chlamydiae species as well as in a subset of Actinobacteria and in the Treponema species. Similarly, in the enzyme UDP-N-acetylglucosamine enolpyruvyl transferase (MurA), which is involved in the synthesis of cell wall peptidoglycan, a 16-aa conserved insert is specifically present in various sequenced chlamydiae and a subset of actinobacteria (i.e., Streptomyces, Actinomyces, Tropheryma, Bifidobacterium, Leifsonia, Arthrobacter, and Brevibacterium). To determine the phylogenetic depths of the GlyA and MurA inserts, the fragments of these genes from two chlamydiae-like species, Simkania negevensis and Waddlia chondrophila, were PCR amplified and sequenced. The presence of the corresponding inserts in both these species strongly indicates that these inserts are distinctive characteristics of the Chlamydiales order. In phylogenetic trees based on GlyA and MurA protein sequences, the chlamydiae species (and also the Treponema species in the case of GlyA) branched with a high affinity with various insert-containing actinobacteria within a clade of other actinobacteria. These results provide strong evidence that the shared presence of these indels in these bacteria is very likely a consequence of ancient lateral gene transfers from actinobacteria to chlamydiae. Pairwise sequence identity and the branching pattern of the GlyA homologues in the phylogenetic tree indicates that the glyA gene was initially transferred from an actinobacteria to an ancestor of the Treponema genus and from there it was acquired by the common ancestor of the Chlamydiales. [Reviewing Editor: Dr. Siv Andersson]     

The Frequency of Eubacterium-to-Eukaryote Lateral Gene Transfers Shows Significant Cross-Taxa Variation Within Amoebozoa

Single-celled bacterivorous eukaryotes offer excellent test cases for evaluation of the frequency of prey-to-predator lateral gene transfer (LGT). Here we use analysis of expressed sequence tag (EST) data sets to quantify the extent of LGT from eubacteria to two amoebae, Acanthamoeba castellanii and Hartmannella vermiformis. Stringent screening for LGT proceeded in several steps intended to enrich for authentic events while at the same time minimizing the incidence of false positives due to factors such as limitations in database coverage and ancient paralogy. The results were compared with data obtained when the same methodology was applied to EST libraries from a number of other eukaryotic taxa. Significant differences in the extent of apparent eubacterium-to-eukaryote LGT were found between taxa. Our results indicate that there may be substantial inter-taxon variation in the number of LGT events that become fixed even between amoebozoan species that have similar feeding modalities. Electronic Supplementary Material Electronic Supplementary material is available for this article at and accessible for authorised users. [Reviewing Editor: Martin Kreitman]     

Effects of Tagetes patula on Active and Inactive Stages of Root-Knot Nematodes

Although marigold (Tagetes patula) is known to produce allelopathic compounds toxic to plant-parasitic nematodes, suppression of Meloidogyne incognita can be inconsistent. Two greenhouse experiments were conducted to test whether marigold is more effective in suppressing Meloidogyne spp. when it is active rather than dormant. Soils infested with Meloidogyne spp. were collected and conditioned in the greenhouse either by 1) keeping the soil dry (DRY), 2) irrigating with water (IRR), or 3) drenching with cucumber (Cucumis sativus) leachate (CL) for 5 wk. These soils were then either planted with cucumber, marigold or remained bare for 10 wk. Suppression of nematode by marigold was then assayed using cucumber. DRY conditioning resulted in the highest number of inactive nematodes, whereas CL and IRR had higher numbers of active nematodes than DRY. At the end of the cucumber bioassay, marigold suppressed the numbers of Meloidogyne females in cucumber roots if the soil was conditioned in IRR or CL, but not in DRY. However, in separate laboratory assays, marigold root leachate slightly reduced M. incognita J2 activity but did not reduce egg hatch (P > 0.05). These finding suggest that marigold can only suppress Meloidogyne spp. when marigold is actively growing. This further suggests that marigold will more efficiently suppress Meloidogyne spp. if planted when these nematodes are in active stage.     

Differential Response to Root-Knot Nematodes in Prunus Species and Correlative Genetic Implications

Responses of 17 Prunus rootstocks or accessions (11 from the subgenus Amygdalus and 6 from the subgenus Prunophora) were evaluated against 11 isolates of Meloidogyne spp. including one M. arenaria, four M. incognita, four M. javanica, one M. hispanica, and an unclassified population from Florida. Characterization of plant response to root-knot nematodes was based on a gall index rating. Numbers of females and juveniles plus eggs in the roots were determined for 10 of the rootstocks evaluated against one M. arenaria, one M. incognita, one M. javanica, and the Florida isolate. These 10 rootstocks plus Nemaguard and Nemared were retested by growing three different rootstock genotypes together in containers of soil infested individually with each of the above four isolates. Garfi and Garrigues almonds, GF.305 and Rutgers Red Leaf peaches, and the peach-almond GF.677 were susceptible to all isolates. Differences in resistance were detected among the other rootstocks of the subgenus Amygdalus. The peach-almond GF.557 and Summergrand peach were resistant to M. arenaria and M. incognita but susceptible to M. javanica and the Florida isolate. Nemaguard, Nemared, and its two hybrids G x N no. 15 and G x N no. 22 were resistant to all but the Florida isolate. In the subgenus Prunophora, Myrobalan plums P.1079, P.2175, P.2980, and P.2984; Marianna plum 29C; and P. insititia plum AD.101 were resistant to all isolates. Thus, two different genetic systems of RKN resistance were found in the subgenus Amygdalus: one system acting against M. arenaria and M. incognita, and another system also acting against M. javanica. Prunophora rootstocks bear a complete genetic system for resistance also acting against the Florida isolate. The hypotheses on the relationships between these systems and the corresponding putative genes of resistance are presented.     

The Genome of Nectria haematococca: Contribution of Supernumerary Chromosomes to Gene Expansion

The ascomycetous fungus Nectria haematococca, (asexual name Fusarium solani), is a member of a group of >50 species known as the “Fusarium solani species complex”. Members of this complex have diverse biological properties including the ability to cause disease on >100 genera of plants and opportunistic infections in humans. The current research analyzed the most extensively studied member of this complex, N. haematococca mating population VI (MPVI). Several genes controlling the ability of individual isolates of this species to colonize specific habitats are located on supernumerary chromosomes. Optical mapping revealed that the sequenced isolate has 17 chromosomes ranging from 530 kb to 6.52 Mb and that the physical size of the genome, 54.43 Mb, and the number of predicted genes, 15,707, are among the largest reported for ascomycetes. Two classes of genes have contributed to gene expansion: specific genes that are not found in other fungi including its closest sequenced relative, Fusarium graminearum; and genes that commonly occur as single copies in other fungi but are present as multiple copies in N. haematococca MPVI. Some of these additional genes appear to have resulted from gene duplication events, while others may have been acquired through horizontal gene transfer. The supernumerary nature of three chromosomes, 14, 15, and 17, was confirmed by their absence in pulsed field gel electrophoresis experiments of some isolates and by demonstrating that these isolates lacked chromosome-specific sequences found on the ends of these chromosomes. These supernumerary chromosomes contain more repeat sequences, are enriched in unique and duplicated genes, and have a lower G+C content in comparison to the other chromosomes. Although the origin(s) of the extra genes and the supernumerary chromosomes is not known, the gene expansion and its large genome size are consistent with this species'' diverse range of habitats. Furthermore, the presence of unique genes on supernumerary chromosomes might account for individual isolates having different environmental niches.     

Host Movement and the Genetic Structure of Populations of Parasitic Nematodes

Mitochondrial DNA (mtDNA) sequence data were used to compare the population genetic structures of five species of parasitic nematodes from three different hosts: Ostertagia ostertagi and Haemonchus placei from cattle, H. contortus and Teladorsagia circumcincta from sheep, and Mazamastrongylus odocoilei from white-tailed deer. The parasites of sheep and cattle showed a pattern consistent with high gene flow among populations. The parasite of deer showed a pattern of substantial population subdivision and isolation by distance. It appears that host movement is an important determinant of population genetic structure in these nematodes. High gene flow in the parasites of livestock also indicates great opportunity for the spread of rare alleles that confer resistance to anthelmintic drugs. All species, including the parasite of deer, had unusually high within-population diversities (averages of 0.019-0.027 substitutions per site between pairs of individuals from the same population). Large effective population sizes (Ne), perhaps in combination with rapid mtDNA evolution, appear to be the most likely explanation for these high within-population diversities.     

Cladistic Approaches in the Study of Soil and Plant Parasitic Nematodes

Cladistic analysis of free-living soil nematodes of the Leptonchoidea(Nematoda: Dorylaimida) resulted in groupings different fromthose obtained by traditional methods. We can interpret distributionsof species groups obtained by phyletic analysis in relationto plate tectonic events. Similar techniques are applicableto plant parasitic nematodes. Grouping on the basis of synapomorphiesproduced a cladogram of genera of the family Heteroderidae (Nematoda:Tylenchida) in which Meloidodera and Cryphodera appear to bethe most ancestral genera and the cyst forming genera the mostderived. A cladogram of groups of species in Heterodera sensulato showed a major division, with the round cyst nematodesand the Cacli group in one grouping and the rest of the Heteroderaspecies in the second. I interpret present-day distributionsby a strict vicariance view and suggest potential falsifiers;and also discuss ancient dispersal routes as alternative waysof thinking about nematode distribution.     

A Satellite Explosion in the Genome of Holocentric Nematodes

Centromere sequences in the genome are associated with the formation of kinetochores, where spindle microtubules grow in mitosis. Centromere sequences usually have long tandem repeats (satellites). In holocentric nematodes it is not clear how kinetochores are formed during mitosis; they are distributed throughout the chromosomes. For this reason it appeared of interest to study the satellites in nematodes in order to determine if they offer any clue on how kinetochores are assembled in these species. We have studied the satellites in the genome of six nematode species. We found that the presence of satellites depends on whether the nematode chromosomes are holocentric or monocentric. It turns out that holocentric nematodes are unique because they have a large number of satellites scattered throughout their genome. Their number, length and composition are different in each species: they apparently have very little evolutionary conservation. In contrast, no scattered satellites are found in the monocentric nematode Trichinella spiralis. It appears that the absence/presence of scattered satellites in the genome distinguishes monocentric from holocentric nematodes. We conclude that the presence of satellites is related to the holocentric nature of the chromosomes of most nematodes. Satellites may stabilize a higher order structure of chromatin and facilitate the formation of kinetochores. We also present a new program, SATFIND, which is suited to find satellite sequences.     

Histologial Studies on the Root-Knot Nematodes Nonresistant and Resistant Root Apex of Humulus lupulus L.

The nematodes (Meloidogy incognita Chitwood.) invade near the root apex of the hop (Humulus lupulus L.), and usually lie near the meristematic zone. Fallowing their entrance, there are the formation of the giant cells and the consequent continuous divisions and activities of the flat meristematic cells, which result in formation of the nematode galls. Their sizes are also related to the number of nematodes. String beads nematode galls are often formed at the nematode-non resistant root part of the "Comet". The process of nematodes entering and later changing of the cells and tissue is almost the same as the formation of the nematode gall in other plants. Fewer nematodes can also enter the root tip of the highly resistant "Qingdao Dahua", but the subsequent special differentiation of the cells and tissues in the root have greatly restrained the activities of nematodes and ultimately to their death.     

Response of Plant Parasitic and Free Living Soil Nematodes to Composted Animal Manure Soil Amendments

In an outside pot experiment, dry pig manure processed on pine sawdust litter and fermented for seven days by house fly larvae (fermented manure), and pine sawdust applied alone, and in combination with a spring application of inorganic nitrogen fertilizer were used to determine their effects on plant parasitic and free-living soil nematodes on sugar beets (cv. Antek). Non amended soil was used as a control. All treatments with fermented pig manure and sawdust with nitrogen fertilizer decreased number of plant parasitic nematodes and also root-fungal feeding nematodes compared to the untreated control. Sawdust applied alone had no effect on plant parasitic and root-fungal feeding nematode suppression. Free-living nematodes which were mainly bacteriovores and fungivores were significantly more abundant in soil amended with fermented pig manure, while the sawdust had no effect on these nematodes. The effect of all tested treatments on omnivores-predators was rather random, and in general, the number of these nematodes decreased after soil amendment applications compared to the untreated control.     

Relationships of Plant Parasitic Nematodes to Sites in Native Iowa Prairies

Soil samples were collected from three native Iowa prairies and analyzed for plant paiasitic nematodes and selected soil properties. Sites or nematodes were clustered with similarities related to habitat by a cluster analysis of site by nematode species and of nematodes by site. Some nematodes occurred in a wide range of prairie habitats, whereas others were more restricted. For example, greater numbers of Xiphinema americanum were in the low, well-drained sites than in the low wet sites or upland dry sites. Wet sites contained fewer nematodes than well-drained sites. Well-drained sites contained mainly Tylenchorhynchus maximus, Helicotylenchus pseudorobustus, and X. americanum. Wetter sites contained almost exclusively X. chambersi, H. hydrophilus, Telylenchus joctus, and an undescribed species of Tylenchorhynchus.     

Occurrence of Gasterophilus intestinalis and some Parasitic Nematodes of Horses in Sweden

A survey was performed on the occurrence of some internal parasites in 461 horses (1–30 years old) slaughtered from October 1992 to September 1993 at the Linköping abattoir in central Sweden. Macroscopical examination was carried out specifically for parasites of the tear ducts and conjunctival sacs of the eyes, and of selected parts of the alimentary tract and car-dio-vascular system. The following parasites were found in selected parts of the large intestine: encapsulated cyathostome larvae (in 35.6% of the horses), and mature strongyle worms (17.4%); in the stomach: Gasterophilus intestinalis (12.3% during October-June) and Habronema muscae (1.1%); and in the conjunctival sac: Thelazia lacrymalis (3.1%). Significantly more encapsulated cyathostome larvae were found during January to June than during other times of the year, and horses aged 1–5 years harboured significantly more larvae than older horses. Severe damage to the cranial mesenteric artery and its main branches was noticed in 16 (6.1%) out of 263 horses specifically examined. However, the 4th stage larva of Strongylus vulgaris was only recovered in 6 (2.3%) of the horses. Quantitative and qualitative faecal egg counts were done on 412 and 384 of the horses, respectively. Eggs of strongyles, Parascaris equorum and Strongyloides westeri were found in 78.1%, 1.9% and 0.2% of the faecal samples, respectively. Highest prevalence of strongyle eggs was found during July to September. Third stage larvae of the following nematodes were recovered from faecal cultures: subfamily Cyathostomum sensu lato (78.1%) Triodontophorus spp. (6.5%), Strongylus vulgaris (3.6%), Trichostrongylus axei (1.3%), while Gyalocephalus sp., Oesophagodontus sp., Poteriostomum sp. and Strongylus edentatus each comprised less than 0.5%.     

Effect of Tillage and Water Management on the Population Behaviour of Root-Knot Nematodes Meloidogyne Triticoryzae in Rice Crop

The effects of puddling and water regimes on hydraulic conductivity (cm/day) of soil and on bulk density (mg/m 3 ) during rice culture, soil physical characteristics of the experimental field population densities of plant-parasitic nematodes have been studied. Puddling reduced the bulk density of soil and decreased the hydraulic conductivity in the upper layers but not in the deeper layers of soil aeration was reduced due to high moisture levels retained in the puddled soil. Population density of M. triticoryzae declined in puddled soil. The invasion of the roots by the second-generation infective juveniles was reduced. The population density of the root-knot nematodes was higher in the non-puddled soil especially in unsubmerged condition compared to puddled and submerged soil. However, if the seedlings were already infected before transplanting and submergence, the nematode could survive well and reproduce within the aerenchyma of the root.     

Comparison of Sequences from the Ribosomal DNA Intergenic Region of Meloidogyne mayaguensis and Other Major Tropical Root-Knot Nematodes

The unusual arrangement of the 5S ribosomal gene within the intergenic sequence (IGS) of the ribosomal cistron, previously reported for Meloidogyne arenaria, was also found in the ribosomal DNA of two other economically important species of tropical root-knot nematodes, M, incognita and M. javanica. This arrangement also was found in M. hapla, which is important in temperate regions, and M. mayaguensis, a virulent species of concern in West Africa. Amplification of the region between the 5S and 18S genes by PCR yielded products of three different sizes such that M. mayaguensis could be readily differentiated from the other species in this study. This product can be amplified from single juveniles, females, or egg masses. The sequences obtained in this region for one line of each of M. incognita, M. arenaria, and M. javanica were very similar, reflecting the close relationships of these lineages. The M. mayaguensis sequence for this region had a number of small deletions and insertions of various sizes, including possible sequence duplications.