THE POTATO TUBER NEMATODE, DITYLENCHUS DESTRUCTOR THORNE, 1945; THE CAUSE OF EELWORM DISEASE IN BULBOUS IRIS

For about twenty-five years a fairly widespread disease of bulbous iris has been known, the cause of which has always been attributed to a biologic race of the stem eelworm Ditylenchus dipsaci (Kühn, 1857), Filipjev, 1936.
Thorne (1945) showed that the nematode causing rot in potato tubers is different from D. dipsaci and named it D. destructor. This nematode has since been reported from Mentha arvensis L. by Hurst (1948) and from Sonchus arvensis L. by Goodey & Goodey (1949).
Examination of nematodes from diseased iris bulbs showed them to possess rounded tail tips and six incisures on each lateral field; characters by which Ditylenchus destructor is distinguished from D. dipsaci.
Cross-inoculation experiments showed that the eelworm causing disease in potato tubers would invade and set up characteristic symptoms in iris bulbs and, in the opposite direction, the eelworm responsible for disease in iris bulbs would give rise to characteristic symptoms in potato tubers. Transfer was also effected from potato and iris to Mentha arvensis and from iris to Sonchus arvensis.
The history of the disease in bulbous iris is briefly reviewed and the biology of Ditylenchus destructor discussed and compared with that of D. dipsaci.
The conclusion that D. destructor is the nematode causing eelworm disease of bulbous iris has been reported earlier (Goodey, J. B. 1950).     

Ultrastructural changes during recovery from anabiosis in the plant parasitic nematode, Ditylenchus

Ultrastructural changes after desiccation and rehydration of the anabiotic fourth-stage juveniles of the plant parasitic nematode Ditylenchus dipsaci (Kuhn) Filipjev are described and quantified. Anabiotic juveniles retain their structural integrity, although the cuticle decreases in thickness and the muscle cell sarcoplasm condenses. In contrast the structure of the non-anabiotic nematode Panagrellus silusae is completely disorganized by desiccation. Following rehydration of D. dipsaci there is a lag phase of 2-3 hr before the nematodes become active. During this period the juveniles undergo an ordered series of morphological changes. The lipid droplets within the intestinal cells coalesce and the cuticle increases in thickness. The muscle cell sarcoplasm expands, the spacing of the thick myofilaments increases and the mitochondria swell before recovering a more normal appearance. These morphological changes, together with earlier metabolic studies, indicate that repair occurs during the lag phase prior to recovery. This may involve membrane repair and the re-establishment of the ionic gradients essential for normal muscle and nerve function.     

Defaecation in the stem nematode, Ditylenchus dipsaci

The process of defaecation in the stem nematode, Ditylenchus dipsaci , has been analysed using cinemicrography. During feeding regular shortenings of the hind end of the body are accompanied by groups of a few defaecation periods. Each period begins as the shortened hind body re-elongates, and consists of several defaecation cycles. In each cycle the posterior rectum becomes dorsally bowed and, as it straightens, the rectal valve, rectum and anus open. Faeces pass out irregularly while the rectum remains open and are forcibly expelled as it closes. Defaecation in D. dipsaci is intermediate between that of the high-pressure nematode Aphelenchoidzs blastophthorus and the low-pressure passive feeder Hexatylus viviparus. We suggest that the main function of defaecation in plant nematodes is to excrete excess water.     

Feeding of the stem nematode, Ditylenchus dipsaci on leaf tissue of field bean, Vicia faba

The behaviour of Ditylenchus dipsaci before, during and after feeding on leaf tissue of Vicia faba is described. Short-term reactions of the food cells and functions of different regions of the pharynx are described and discussed in relation to the changing pressure relationships between nematode and cell.     

Nematode resistance in plants: the battle underground

Parasitic nematodes infect thousands of plant species, but some plants harbor specific resistance genes that defend against these pests. Several nematode resistance genes have been cloned in plants, and most resemble other plant resistance genes. Nematode resistance is generally characterized by host plant cell death near or at the feeding site of the endoparasitic worm. The timing and localization of the resistance response varies with the particular resistance gene and nematode interaction. Although there is genetic evidence that single genes in the nematode can determine whether a plant mounts a resistance response, cognate nematode effectors corresponding to a plant resistance gene have not been identified. However, recent progress in genetics and genomics of both plants and nematodes, and developments in RNA silencing strategies are improving our understanding of the molecular players in this complex interaction. In this article, we review the nature and mechanisms of plant-nematode interactions with respect to resistance in plants.     

OBSERVATIONS ON THE ATTACK BY THE STEM EELWORM, DITYLENCHUS DIPSACI, ON STRAWBERRY

Four biologic races of Ditylenchus dipsaci can cause stem eelworm disease on twelve varieties of strawberry, all but one cultivated in Great Britain. The course of attack on strawberry, host reaction and the ways in which the disease originates and is disseminated are described.     

DNA polymorphism in the stem nematode Ditylenchus dipsaci: development of diagnostic markers for normal and giant races.

DNA polymorphism in the Ditylenchus dipsaci complex was investigated using amplified fragment length polymorphism (AFLP) to determine the relationships among populations growing mainly on Vicia faba and to develop diagnostic markers. Twenty-two populations of D. dipsaci originating from different geographical areas and one population of Ditylenchus myceliophagus were used. AFLP proved to be a powerful method to reveal intraspecific polymorphism even within the giant type. The analysis showed a clear distinction between the giant and normal populations, with genetic distances similar to those observed between normal populations and D. myceliophagus or giant populations and D. myceliophagus, strengthening the hypothesis that these two nematode types could be considered distinct species. Two specific AFLP markers differentiating the two types were converted into sequenced characterized amplified region (SCAR) markers. Used in a multiplex PCR, the SCAR primers proved to be a rapid and efficient tool to identify the giant and the normal types of D. dipsaci.     

Population increase of Ditylenchus dipsaci (Kühn) in the narcissus and the spread of the nematode through the soil

A bioassay technique, using forty onion seedlings per pot, was used to determine the soil population of the narcissus race of Ditylenchus dipsaci .
In experiments with narcissus, a progressive increase of D. dipsaci populations was shown, reaching a peak at the end of the growing season. This rise in population in May/June was associated with a rise in temperature. Increase in inoculum gave a significantly larger population in the foliage but not in the bulbs. Nematodes moved actively and were transported passively both up and down within the host plant. The presence of spikkels was associated with nematodes in the active intercalary meristem.
Migration from the plant into the soil and back into the plant, mostly via the base of the bulb, was continuous throughout the growing season and related to the population increase within the plant. Migration of nematodes from wetted dried leaf tissue continued over 60 days. Spread of the nematode through the soil was slow in the absence of external agencies, such as water run-off and methods of cultivation.     

Cryopreservation of Radopholus similis, a tropical plant-parasitic nematode

For obligate plant-parasitic nematodes, cryopreservation has advantages over the usual preservation methods on whole plants or axenic culture systems, because the latter two are labourious and time and space consuming. In addition, cross contamination among different isolates can occur easily. Moreover, specific genetic studies require maintenance of the original population. The nematode under investigation, Radopholus similis, is a plant-parasitic nematode from the humid tropics. Therefore, any treatment at low temperatures is likely to add extra stress to the nematode, making the development of a cryopreservation protocol extremely difficult. In this paper, we describe experiments to achieve a successful cryopreservation protocol for the tropical nematode R. similis using vitrification solution-based methods based on a well defined mixture of cryoprotectants in combination with ultra-rapid cooling and thawing rates. A two-step treatment was used consisting of an incubation in glycerol followed by the application of a vitrifying mixture of methanol, glycerol and glucose. After cryopreservation, the pathogenicity of the nematodes was not altered, since they could infect and reproduce on carrot discs after recovery in the Ringer solution. The cryopreservation method described can be used for routine cryopreservation of R. similis lines from different origins.     

Compatibility Between Pasteuria penetrans Isolates and Meloidogyne Populations from Spain

Several populations of Pasteuria isolated from fields in Spain were compared with other Pasteuria populations, held in collections at the Institute de Recerca i Tecnologia, Agroalimentaries (IRTA), Cabrils or IACR-Rothamsted, for their ability to adhere to and infect root-knot nematodes ( Meloidogyne spp.) grown on host plants differing in their susceptibility to root-knot nematodes. The results showed a high level of variation in both the ability of a population of Pasteuria to adhere to a particular population of nematode and vice versa. In particular the isolates of Pasteuria originating from M. hapla retained a high level of specificity for the species from which they originated. The infection of the nematodes by the bacteria was generally low, even when nematodes were encumbered with relatively high levels of spores. It is suggested that prolonged storage (6 years) may reduce the ability of spores to infect nematodes independently of adhesion.     

The occurrence of Anguillulina dipsaci (Kühn.) on weed hosts, including new host records in fields of oats affected by 'tulip-root'

The examination of eighteen weeds in three fields of oats affected with 'tulip-root' is described. Cleavers, common chickweed, mouse-ear chickweed and sandwort, were found to be hosts of A. dipsaci , the last two weeds being new host records. Successful transferences of A. dipsaci have been made from chickweed to oats and from oats back to chickweed. Transference of A. dipsaci has also been made from cleavers and sandwort to oats.     

Water uptake and morphological changes during recovery from anabiosis in the plant-parasitic nematode, Ditylenchus dipsaci

The changes in water content and structural organization that occur during the rehydration of the anabiotic 4th-stage juveniles of the plant-parasitic nematode Ditylenchus dipsaci are described. Morphological changes include the coalescence of lipid droplets and the appearance of a hyaline layer beneath the cuticle. Changes in diameter are mainly responsible for volume changes. Changes in the appearance of the hyaline layer suggests that the muscle cells undergo some sort of repair process before activity resumes.     

Ultrastructural changes during desiccation of the anhydrobiotic nematode Ditylenchus dipsaci

Ultrastructural changes during desiccation of the anhydrobiotic nematode Ditylenchus dipsaci were followed and quantified after preparation of material at different levels of hydration using freeze substitution techniques. Some shrinkage was caused by processing in the more hydrated specimens but the changes observed correspond to those observed in live nematodes by light microscopy, indicating that the technique is useful for following changes during desiccation. The overall pattern of changes was a rapid decrease in the magnitude of the measured parameter during the first 5 min of desiccation, followed by a slower rate of decrease upon further desiccation. This was observed in the cuticle, the lateral hypodermal cords and the muscle cells and is consistent with the pattern of water loss of the nematode. The contractile region of the muscle cells, however, proved an exception and the muscle fibres appear to resist shrinkage and packing until water loss becomes severe. The mitochondria swell and then shrink during desiccation, which may indicate disruption of the permeability of the mitochondrial membrane. A decrease in the thickness of the cortical zone was the most prominent change in the cuticle and this may be related to the permeability slump which occurs during the first 5 min of desiccation.     

Wolbachia genomes: revealing the biology of parasitism and mutualism

Wolbachia bacteria are endosymbiotic partners of many animal species, in which they behave as either parasites (in arthropod hosts) or mutualists (in nematode hosts). What biochemistry and biology underpin these diverse lifestyles? The recent complete sequencing of genomes from Wolbachia that infect the arthropod Drosophila melanogaster and the nematode Brugia malayi, together with the partial genome sequencing of three Wolbachia strains found in drosophilids, enables this question to begin to be addressed. Parasitic arthropod Wolbachia are characterized by the presence of phages that carry ankyrin-repeat proteins; these proteins might be exported to the host cell to manipulate reproduction. In nematode Wolbachia, which lack these phages, several biochemical pathways can deliver essential metabolites to the nematode hosts. Nematode Wolbachia might also have a role in modulating the mammalian host immune system but the sequenced Wolbachia genomes lack the genes to synthesize lipopolysaccharide, raising questions about the nature of the inducing molecule. The Wolbachia surface protein might carry out this function.     

续断对兔骨折愈合过程中相关基因表达和血钙、磷含量的影响

本研究通过检测中药材续断对家兔骨折模型愈合过程中与成骨密切相关基因的表达,以及血清中钙、磷含量变化,探讨其对骨折愈合的促进机制。构建家兔骨折缺损模型,术后按组分别给予续断和蒸馏水灌胃,并分别检测骨保护素(OPG)、骨保护素配体(OPGL)、局部转化生长因子β1(TGF-β1)和骨形态发生蛋白-2(BMP-2)的基因表达以及血清中Ca、P、碱性磷酸酶(ALP)含量。结果表明,续断治疗组中血钙、血磷和ALP的含量在灌胃第2周,第3周和第4周后均有明显升高,且在第三周时达到最大值。同时,OPG、TGF-β1、BMP-2三个基因在用续断治疗的不同时期呈现不同程度的表达上调,OPGL则在治疗早期表达下调。推测续断对骨折的治疗可能是通过调控OPG、OPGL、TGF-β1、BMP-2等基因在骨愈合不同阶段的表达量和血清中Ca、P、ALP的含量来促进骨骼生长。     

Changes in surface features during desiccation of the anhydrobiotic plant parasitic nematode Ditylenchus dipsaci

The anhydrobiotic nematode Ditylenchus dipsaci is a fast-dehydration strategist, itself generating the slow rate of water loss necessary for survival. A permeability slump occurs during the initial phases of desiccation. This may be produced by changes in the nematode's cuticle. Two scanning electron microscopic techniques were used to follow changes in surface structures during desiccation. Freeze substitution and critical-point drying produced artifacts that obscured changes produced by the desiccation of the nematode. Low-temperature field emission scanning electron microscopy (FESEM) was successful in following changes that reflected those observed by light microscopy (LM). Significant changes in diameter, the lateral alae, and the cuticular annulations were demonstrated using this technique. Two types of annulations were observed: the major annulations, which extended to meet the margins of the lateral alae, and the minor annulations, which did not. With desiccation the prominence of the annulations increased, their spacing decreased, and the minor annulations extended closer to the margins of the lateral alae. These observations are consistent with the permeability slump resulting from a decrease in the width of the annulation groove and an increase in its depth. However, this requires confirmation using techniques that can follow annulation changes in individual nematodes.     

Transmission of the hop strain of arabis mosaic virus by Xiphinema diversicaudatum*

Hop plants became infected with the hop strain of arabis mosaic virus (AMV(H)) when grown in hopfield and woodland soil in which infected plants had been growing. Infection occurred in soil infested with the dagger nematode Xiphinema diversicaudatum, but neither in uninfested soil nor in soil previously heated to kill nematodes. X. diversicaudatum transferred direct from hop soils transmitted AMV(H) to young herbaceous plants and to hop seedlings; some of the hop seedlings developed nettlehead disease. A larger proportion of plants was infected using X. diversicaudatum obtained from a woodland soil and then given access to the roots of hop or herbaceous plants infected with AMV(H). AMV(H) was transmitted by adults and by larvae, in which the virus persisted for at least 36 and 29 wk, respectively. Difficulties were encountered in detecting AMV(H) in infected hop plants, due partly to the delay in virus movement from roots to shoots. Infection of hop shoots was seldom detected until the year after the roots were infested and sometimes nettlehead symptoms did not appear until the third year. Isolates of arabis mosiac virus from strawberry did not infect hop. The results are discussed in relation to the etiology and control of nettlehead and related diseases of hop.     

A SNARE-Like Protein and Biotin Are Implicated in Soybean Cyst Nematode Virulence

Phytoparasitic nematodes that are able to infect and reproduce on plants that are considered resistant are referred to as virulent. The mechanism(s) that virulent nematodes employ to evade or suppress host plant defenses are not well understood. Here we report the use of a genetic strategy (allelic imbalance analysis) to associate single nucleotide polymorphisms (SNPs) with nematode virulence genes in Heterodera glycines, the soybean cyst nematode (SCN). To accomplish this analysis, a custom SCN SNP array was developed and used to genotype SCN F3-derived populations grown on resistant and susceptible soybean plants. Three SNPs reproducibly showed allele imbalances between nematodes grown on resistant and susceptible plants. Two candidate SCN virulence genes that were tightly linked to the SNPs were identified. One SCN gene encoded biotin synthase (HgBioB), and the other encoded a bacterial-like protein containing a putative SNARE domain (HgSLP-1). The two genes mapped to two different linkage groups. HgBioB contained sequence polymorphisms between avirulent and virulent nematodes. However, the gene encoding HgSLP-1 had reduced copy number in virulent nematode populations and appears to produce multiple forms of the protein via intron retention and alternative splicing. We show that HgSLP-1 is an esophageal-gland protein that is secreted by the nematode during plant parasitism. Furthermore, in bacterial co-expression experiments, HgSLP-1 co-purified with the SCN resistance protein Rhg1 α-SNAP, suggesting that these two proteins physically interact. Collectively our data suggest that multiple SCN genes are involved in SCN virulence, and that HgSLP-1 may function as an avirulence protein and when absent it helps SCN evade host defenses.