CSIRO PUBLISHING

www.publish.csiro.au/journals/app

Australasian Plant Pathology, 2008, 37, 289--297

Australian anguinids: their agricultural impact and control

Ian T. RileyA,B,D and Martin J. BarbettiC
A

Plant and Food Science, School of Agriculture Food and Wine, The University of Adelaide, Waite Campus, Urrbrae, SA 5064, Australia. B Plant and Soil Health, SARDI, Plant Research Centre, Hartley Grove, Urrbrae, SA 5064, Australia. C School of Plant Biology, Faculty of Natural and Agricultural Sciences, The University of Western Australia, Crawley, WA 6009, Australia. D Corresponding author. Email: ian.riley@adelaide.edu.au

Abstract. The life history, importance and management of the anguinid nematodes of Australia are briefly summarised. Anguina funesta is the most important because of its association with the toxigenic actinomycete, Rathayibacter toxicus, and consequent contribution to annual ryegrass toxicity (ARGT). Although study of A. funesta has provided a good understanding of its biology and workable approaches to control, outbreaks of ARGT still occur and issues remain for exports. An undescribed Anguina species, also associated with R. toxicus and livestock deaths (flood plain staggers), is of economic concern but its dispersed and sporadic occurrence has limited impetus and opportunity for its further study. Anguina tritici exists in Australia only as remnant populations, having been eliminated by mechanised seed cleaning and crop rotations over most of its widespread range of a century ago. A further seven anguinids are known from Australia and, although of minimal economic significance, represent an opportunity for description of new species and further study of this highly specialised group of plant-parasitic nematodes. Introduction Three Anguina species (seed-gall nematodes) are or were responsible for economic problems in Australia. Anguina funesta in Lolium rigidum (Price et al. 1979; McKay and Ophel 1993), together with the toxigenic actinomycete (Rathayibacter toxicus), causes annual ryegrass toxicity (ARGT), an ongoing concern for livestock production and oaten hay exports in South Australia (SA) and Western Australia (WA). Anguina sp. in Lachnagrostis filiformis and Polypogon monspeliensis (McKay et al. 1993; Bertozzi and McKay 1995; Bertozzi 2003), also associated with R. toxicus, causes flood plain staggers (FPS), a sporadic poisoning of livestock in New South Wales (NSW) and SA. Anguina tritici in cereals (Southey 1972; Riley 1992), previously found across southern Australia, is now largely an historical problem but persists and causes minor economic impact in WA. In addition, there are several other anguinids (leaf and seed-galls species) that have no direct economic impact but are occasionally of phytosanitary concern for export commodities. This paper summarises the biology, occurrence, impact and control of the three important Anguina species in Australia and catalogues the other anguinids. As part of a compilation of reviews on Australasian nematology, the focus will be the nematodes and their associated microorganisms. In the cases of the corynetoxin (Edgar et al. 1982) poisonings (ARGT and FPS), there is a considerable body of work on the toxicology and development of animal health solutions (e.g. Davies et al. 1993; Edgar 1994; May and Stewart 1998; Than et al. 1998b; Finnie 2006; Lee et al. 2006), such as by stock management, nutrition, antidote and/or vaccine. Also, for ARGT, one approach attempted is to avoid the problem by eliminating and/or replacing the host
Australasian Plant Pathology Society 2008

(Burdass 1986; see Gill 1996a, 1996b and Matthews 1996a, 1996b for reviews of L. rigidum importance, ecology and control). However, these aspects are only mentioned briefly here in an effort to confine the scope of this discussion to the nematology of anguinids in Australia. Anguinids The family Anguinidae belongs to the order Tylenchida that contains stylet-bearing nematodes, many of which feed on plants (Krall 1991). In the Anguinidae there are several genera of seed and leaf gall nematodes, which are highly specialised and obligate parasites that induce galls in various plant tissues (Chizhov and Subbotin 1987; Subbotin et al. 2004). Three anguinid genera, Anguina, Mesoanguina and Subanguina, are recorded in Australia. A. funesta and annual ryegrass toxicity Biology and occurrence A. funesta is a seed-gall nematode that induces galls in the ovaries of Festuca, Lolium and Vulpia species (Price et al. 1979; Riley 1995; Fig. 1a, c). At maturity these galls contain anhydrobiotic second-stage juveniles (J2), the survival and dispersal stage of the nematode and a suitable stage to begin the description of the life cycle (Bird and Stynes 1981a, 1981b, 1981c; McKay et al. 1981; McKay 1985). In the Mediterranean climate of southern Australia, the rains arrive in autumn wetting the galls, which, by this stage, have shattered from the host inflorescence along with the host diaspores and are on the soil surface. The nematodes rehydrate and regain activity as moisture is slowly absorbed through
10.1071/AP08022 0815-3191/08/030289

290

Australasian Plant Pathology

I. T. Riley and M. J. Barbetti

(a)

(b)

(c)

(d )

(e)

(f )

(g)

Fig. 1. (a--c) Diaspores and seeds, nematode galls (dark) and bacterially colonised nematode galls (yellow) in three grass hosts infested with seed-gall nematode, Anguina spp. (a) Lolium rigidum with Anguina funesta and Rathayibacter toxicus; (b) Triticum aestivum with Anguina tritici and Rathayibacter tritici; (c) Vulpia myuros with A. funesta and R. toxicus; (d, e) Dilophospora alopecuri; (d) infected L. rigidum seedheads; (e) conidia adhering to cuticle of A. funesta; (f, g) diaspores, enlarged infested florets, nematode galls (dark) and R. toxicus-colonised nematode galls (yellow) in two hosts infested with the flood plain staggers nematode, Anguina sp.; ( f ) Polypogon monspeliensis (including apical gall, far right); (g) Lachnagrostis filiformis (photographs f and g courtesy of A. McKay).

Australian anguinids

Australasian Plant Pathology

291

persistent floral bracts, gall wall and nematode cuticle. With a sufficient period of soil surface moisture, the surrounding tissues degrade and the nematodes emerge onto the soil surface. If the rainfall is not sufficient and the soil surface dries (as occurs with summer rainfall events), the J2 will return to an anhydrobiotic state until further rain occurs. The J2 can survive through several hydration/dehydration cycles (Preston and Bird 1987). On a moist soil surface, the J2 will actively seek a host plant to invade (Riley and McKay 1991a), probably assisted by rain splash. The J2 congregate at the meristem of infested tillers and wait for ovary initiation before initiating a gall by invading the ovary and modifying its development. Within the gall, J2 feed and moult through two further juvenile stages to adults (females and males, usually a few but up to 20 per gall, Riley and Bertozzi 2004). Sexual reproduction is essential and results in several hundred eggs per gall (Riley and Bertozzi 2004). The nematodes develop to J2 in the eggs and hatch in the gall. The freshly hatched J2 cannot survive desiccation, but mature physiologically to become the survival stage as the host senesces and desiccates in the spring or early summer. The nematode alone would be of minimal economic significance as seed set in L. rigidum is not greatly affected and this grass, although an important pasture species, is regarded as a major weed in southern cropping areas (Gill 1996a). However, the toxigenic R. toxicus adheres to the cuticle of the J2 and is carried into the host inflorescence and nematode galls, where it colonises these structures (Bird and Stynes 1977; Fig. 1a, c). As the host matures, R. toxicus produces potent glycolipid toxins known as corynetoxins (Edgar et al. 1982) that inhibit n-glycosylation of proteins and are often fatal to livestock (Jago et al. 1983). Fig. 1a shows the L. rigidum seed, nematode galls and bacterially colonised galls.

Various factors contribute to the dynamics of invasion and reproduction of A. funesta, including the timing and intensity of autumn rains, time of host germination, host density, interplant competition, grazing frequency and intensity, and degree of bacterial colonisation. Some progress has been made in modelling these interactions (McKay 1985) but a predictive model is still to be developed. Fig. 2 shows the development of the nematode relative to that of the host. For maximum nematode reproduction or bacterial colonisation, and thereby potential of greatest economic impact, it is important that (1) nematode invasion is synchronised with ovary initiation (McKay et al. 1981), and (2) infested tillers are not lost due to grazing or interplant competition or prevented from reaching maturity by grazing or agronomic practice. Since the first outbreaks of ARGT and recognition of its causal organisms in SA (1955, McIntosh et al. 1967) and WA (late 1960s, Gwyn and Hadlow 1971), they have spread across much of the cropping zone in these States (Fig. 3). As the occurrence of problematic population densities of A. funesta will be driven by the factors mentioned above, they occur within rainfall zones that optimise synchronisation of J2 invasion and host development and mostly in fields where L. rigidum is promoted and protected through to maturity by cropping practices. It is still possible that ARGT outbreaks will occur in other areas as A. funesta continues to spread and increase in density. Also, the distribution of A. funesta will always be wider than that of ARGT because not all situations, which enable establishment and persistence, will support high population densities of the nematode. For example, A. funesta has been found at low population densities in Victoria but ARGT outbreaks have not (Riley 1996; Clarke and Nurse 1999). In this case, L. rigidum densities and climatic conditions would appear favourable but A. funesta populations

Life cycle of a seed-gall nematode, Anguina sp. Host

Vegetative Reproductive

Nematode
On soil In plant J2
Moult

In gall J3
Moult

J4
Moult

Adult J1 Egg deposition

Moult in egg

J2 Anhydrobiosis

Gall initiation

Rehydration Emergence

Invasion

Fig. 2. Life cycle of seed-gall Angina species, such as A. funesta and A. tritici.

Hatch

292

Australasian Plant Pathology

I. T. Riley and M. J. Barbetti

Western Australia Anguina tritici

FPS (Anguina sp.) South Australia

New South Wales

ARGT (Anguina funesta)

Victoria

Fig. 3. Map of southern mainland Australia showing the approximate occurrence of Anguina funesta as indicated by annual ryegrass toxicity outbreaks, the flood plain staggers nematode, Anguina sp. and Anguina tritici.

might be regulated by the fungal antagonist, Dilophospora alopecuri (discussed below). A. funesta is not recorded outside Australia. However, molecular phylogenic analysis groups it with anguinids of Euro-Asian origin (Subbotin et al. 2004). It is likely to be exotic to Australia and to have remained unnoticed in its native range because host densities and management do not allow high nematode population densities to develop as they do in L. rigidum in southern Australia. If R. toxicus is not present, then stock poisoning cannot bring attention to infestations of the nematode. Also, the history of the spread of A. funesta and outbreaks of ARGT are consistent with the nematode being an introduced organism (McKay et al. 1985). Agricultural impact The direct impact of A. funesta is livestock deaths and production losses caused by corynetoxins produced by its associated bacterium, R. toxicus. In SA, the number of outbreaks has diminished over the last two decades to only a few per year, but in its peak year ~2500 sheep and 50 cattle were killed by ARGT (McKay and Ophel 1993; McKay and Riley 1993). Outbreaks continue in WA and although detailed records are no longer collected, it is estimated that the long-term average is ~30 000 sheep and several hundred head of cattle killed each year (J. Allen, pers. comm.), a decline from the worst year (1991) when as many as 88 000 sheep died (Roberts et al. 1994). The difficult-to-quantify impacts of A. funesta and ARGT include production losses in livestock with subclinical exposure to corynetoxins (Retallick et al. 2007) and from pastures subjected to control measures. In livestock, subclinical effects on wool and reproduction are known (Davies et al. 1996) and reductions in growth and feed conversion are also suspected. In pastures, the side effects of the common control measures include a decline in pasture productivity and quality as less productive grasses invade and legume seed set and persistence is reduced. Also, landowners hesitate to invest in pasture improvement because the gains might be compromised by a need to control ARGT. Therefore, ARGT can add substantially to management uncertainties.

In addition to effects on livestock, A. funesta and ARGT have other impacts (both direct and indirect) and these are considered to be much greater than the livestock losses. These include the cost of control measures and the side effects of control (discussed below). The other main direct impact is the loss of standing or harvested feed. Pasture, crop stubbles, hay, feed grains and screenings that have caused ARGT, or are sufficiently infested to present a risk of causing ARGT, cannot be used. Production of cereal hay, such as oaten hay, is grown to help control L. rigidum in cropping areas, but it can be potentially toxic if infested L. rigidum content is high (Than et al. 1998a). Outbreaks of ARGT in Japan from imported Australian hay (Nogawa et al. 1997) gave rise to industry-driven, mandatory pre-export testing to ensure R. toxicus concentrations are below the acceptable threshold. An immunoassay for corynetoxin is available (Than et al. 1998a) but is not implemented for routine use at this stage, so R. toxicus is used as an indicator of potential toxicity (Masters et al. 2006). The cost of testing and loss of premium for hay rejected for export is another direct impact. The potential for corynetoxins to enter the food chain is also a consideration (Edgar 2004). To limit the impact of ARGT outbreaks, stockowners need to maintain frequent, even daily, surveillance of livestock during the late spring and summer. Seeing animals poisoned by corynetoxins is highly distressing for stockowners. If the animals represent a significant part of a long-term genetic improvement program, this stress is further exacerbated. In this way, ARGT has far reaching implications; much more than for example, yield losses in crops resulting from plant disease. Control Control of ARGT has largely concentrated on control of A. funesta, either directly or by controlling host populations (Brown and Vogel 1986; Maling and OCallaghan 1992; McKay and Ophel 1993). The weakest point in the life cycle of A. funesta is after infested tillers elongate but before the J2 develop the capacity to survive anhydrobiotically. During this period, killing infested tillers by mowing, grazing or herbicides also kills the adults, eggs and J2 they contain. Reducing the

Australian anguinids

Australasian Plant Pathology

293

nematode population in this way reduces the risk of ARGT in subsequent years. Furthermore, early treatment of infested tillers can prevent R. toxicus producing toxin (toxin production increases as the plant senesces) and reduce the risk of ARGT in the current season. However, timing is critical. Systemic herbicides, such as glyphosate, act slowly and if applied late, their use might not prevent the nematode entering its survival stage. R. toxicus in cut pasture that is re-wet by rain can resume toxin production. Efforts to control A. funesta by reducing seed production by L. rigidum in the previous season or its establishment in the current season by mowing, herbicides or cultivation can reduce the host population density but not necessarily the risk of ARGT. If J2 survival is unaffected, the reduced population of L. rigidum in the next season can be more heavily infested, with greater bacterial development and still represent a serious risk of ARGT. Controlling A. funesta by reducing the host population needs to be a long-term strategy and is best done in conjunction with direct control of the nematode (Riley and Gill 1994). Genetic resistance against A. funesta, consisting of two dominant genes, is available in commercial ryegrass cultivars (e.g. Lolium hybrid cv. Safeguard, Valley Seeds Australia Pty Ltd, Vic., McKay 1993; Anon. 1994, 2004). As L. rigidum is out-crossing, the resistance genes can be incorporated into the progeny of the local ecotype, if flowering coincides with that of the resistant cultivar, to provide incomplete but still useful resistance. The resistant plants not only fail to support A. funesta reproduction but also act as trap plants. Host resistance can deliver good control of A. funesta provided the background population of susceptible ryegrass is relatively small. If a high frequency of the resistance genes enters and persists in the local ryegrass population, this is an attractive option for control, because cv. Safeguard itself might not persist. However, success depends on achieving good initial control of L. rigidum, which is clearly not consistently achievable. A. funesta and other Anguina species are also vectors of the plant pathogenic fungus, D. alopecuri (Gibson and Sutton 1976; Fig. 1d). The conidial appendages adhere to J2 and are carried into the host (Bird and McKay 1987; Riley and McKay 1990; Fig. 1e). Adhesion to J2 and colonisation of the host inflorescence by D. alopecuri inhibits gall formation and bacterial colonisation. The fungus is widespread over much of the range of A. funesta, including Victoria (Riley 1996). The decline of the frequency of ARGT outbreaks in the Great Southern Region of WA is attributed to this fungus (Riley 1994a) and it could also be the reason that ARGT has not occurred in Victoria. The fungus is produced commercially on cracked rice and applied to areas where ARGT is still problematic (Yan and Riley 1998, 2003, 2005; Biological and Resource Technology Pty Ltd, Bull Creek, WA). Based on farmer experience, it is difficult to establish in dry years, but once established, D. alopecuri has provided useful control (G. Yan, pers. comm.). As with most biological control, it is not effective in all circumstances. Nevertheless, this is the only available method of control that does not depend to greater or lesser extent on controlling L. rigidum, making it an important component of integrated control (Barbetti and Riley 2006). Research on non-toxigenic Rathayibacter species that could displace R. toxicus and provide a complementary biological

control to D. alopecuri is warranted (Riley and McKay 1991b; Riley 1994b; Riley and Yan 1998). However, further progress in this area will require quarantine approval for field evaluation, given that the most promising strains are exotic to Australia. Anguina sp. and flood plain staggers Biology and occurrence The Anguina sp. that parasitises L. filiformis and P. monspeliensis (Fig. 1f, g), studied by Bertozzi (2003) and others (Bourke et al. 1992; Bryden et al. 1994; Bertozzi and McKay 1995; Davis et al. 1995), is awaiting formal description and naming (it will be called the FPS nematode in this paper). Although, in many ways its biology is similar to A. funesta, described above, there are important differences, which largely represent adaptations for survival in ecosystems subject to seasonal inundation. The biology of the FPS nematode is well studied in P. monspeliensis (Bertozzi 2003) but not in L. filiformis. In years when pastures in low-lying areas of south-east SA are flooded for 1--5 months, almost pure stands of P. monspeliensis develop. In dry years, barley grass (chiefly Hordeum leporinum), a non-host for the FPS nematode, dominates these pastures. The adaptations of the FPS nematode to flooding and its host include (1) the induction of galls in apical meristems and panicle branches as well as ovary initials, (2) two generations of galls per year, (3) rapid emergence of J2 from rehydrated galls and host invasion, and (4) ability to survive inundation. Following the autumn rains in the south-east of SA, it is vital for the FPS nematode to emerge, invade and induce galls rapidly because flooding can follow in the winter. Under flooded conditions the J2 cannot invade and, although it appears the J2 can survive longer under water than in moist soil, there is still a considerable attrition rate. Thus, the rapid rehydration, emergence and host invasion (24 h) by the FPS nematode is in distinct contrast to A. funesta, which can take a month to complete this phase of its life cycle (Price et al. 1979). Likewise, if the FPS nematode were to wait at the apical meristem to induce galls in ovary initials, as does A. funesta, it would be unlikely to survive the period of inundation. Its attrition rate would be high because under peak flooding, P. monspeliensis is reduced to a single tiller. The rapid formation of apical galls allows the FPS nematode to complete a life cycle by the early winter and provide a new cohort of J2 to invade tillers that form as the flooding recedes. The second cycle of invasion and gall formation that occurs in the spring leads to the production of seed galls and a relatively minor proportion of panicle branch galls. The development of the FPS nematode from J2 to adults within the gall takes up to 14 days as it does with A. funesta and A. tritici (Swarup and Gupta 1971; Price et al. 1979). By the time heads of P. monspeliensis are emerging from the flag leaf sheath, the first J2 are hatching. In contrast to A. funesta, the FPS nematode induces enlargement of infested florets (Fig. 1f ). The final stages of the life cycle of the FPS nematode parallel that of A. funesta, with the mature galls containing anhydrobiotic J2 that survive summer to emerge with the autumn rains of the next year. As with A. funesta, the FPS nematode is a vector for R. toxicus. The bacterium can colonise the three gall types (apical, panicle and seed galls) and the inflorescences, and produces corynetoxins at concentrations lethal to livestock (Edgar et al. 1994).

294

Australasian Plant Pathology

I. T. Riley and M. J. Barbetti

Little is known from field observations of the FPS nematode in L. filiformis other than it forms apical and seed galls, is associated with R. toxicus and high population densities evidently only occur after several seasons with winter rainfall or floods (conditions that have not reoccurred since its first detection in NSW in 1991). It is likely that the apical galls are important for the survival in a host that matures rapidly following infrequent and transient flooding. The FPS nematode is most common in seasonally inundated pasture in the south-east of SA (Fig. 3), being found in 36% of 265 sites where P. monspeliensis occurred in a 1991--92 survey (Bertozzi 2003). It was not found in host populations along the Murray River in SA in 1991, but a single population of infested P. monspeliensis was found in northern NSW in 1993 (Fig. 3). In L. filiformis, it has only been recorded in the catchment of the Darling River upstream from Bourke, NSW (Fig. 3). Most infested sites were found following outbreaks of FPS in livestock in 1991. Planned and ad hoc surveys in NSW between 1993 and 1996 found that the nematode and bacterium were more widely spread in the upper catchment of the Darling River than indicated by the FPS outbreaks. Despite the overlapping distribution of the hosts, the populations of the FPS nematode in NSW and SA appear largely host specific. However, it is not known if this is due to physiological adaptation or a consequence of the interaction of climatic conditions and host phenology. Agricultural impact The accumulated impact of FPS is much less than that of ARGT, but nevertheless unpredictable, and untreatable poisoning of farm animals will always be a significant concern for stockowners. There are no reliable data on livestock deaths in the south-east of SA from FPS, but deaths and reduced fecundity over more than 30 years are now attributed to FPS. In 1990, a particularly bad year, total losses of 2000 sheep and 40 cattle were attributed to FPS in SA (C. Trengrove, cited as a pers. comm. in McKay et al. 1993). Over the last decade or so, changes in land use, drainage works and dry seasons mean that FPS is now rare in SA. In NSW, deaths of 1722 cattle, 2466 sheep and 11 horses in 1991 were the first and major recognised impact of FPS (Davis et al. 1995). Again, with a lack of successive wet winters and the return of average to below-average rainfall has meant that FPS outbreaks are rare events in northern NSW. Control The sporadic character of FPS nematode and the inundation of pastures make direct control of the nematode difficult. The apical galls of the FPS nematode mean that some methods used for A. funesta are unlikely to be successful because they depend on the vulnerability of pre-survival stage nematodes in galls in the seedhead. Bertozzi (2003) found that promotion of a canopy of clover in the spring would reduce the contribution of apical galls in stunted tillers to the nematodes survival. D. alopecuri occurs in the south-east of SA and might provide useful biological control including the nematodes in apical galls. No control for the FPS nematode in L. filiformis has been proposed and risk monitoring though surveillance for infestations in extensive rangelands is unlikely to be attractive to stockowners.

For now, the decline in host prevalence in both regions has removed the impetus for the development of control measures, but stockowners and advisors should remain aware of the risk if conditions favourable to the FPS nematode reoccur. A. tritici Biology and occurrence A. tritici, a seed-gall nematode of wheat and some other cereals, has a life cycle similar to that of A. funesta. A. tritici galls (Fig. 1b) are induced by more J2, are larger and contain many more progeny than galls of A. funesta (Riley and Bertozzi 2004). Symptoms of infestation are more obvious, with damage caused to the vegetative apex by feeding/probing of J2 resulting in distorted leaves, tillers and seedheads (Southey 1972). Key survival and spread of A. tritici is in crop seed and no naturalised hosts are known in Australia. This leads to a significantly different dynamic than seen in A. funesta, with its widely naturalised and difficult-to-manage weed as the major host. A. tritici also acts as a vector for a bacterium, the non-toxigenic Rathayibacter tritici, which causes gummosis in the host and increases the impact of the nematode on yield (Riley and Reardon 1995). Galls can also be colonised (Fig. 1b). The nematode is also a potential vector of R. toxicus (Riley 1992) and a recorded vector of D. alopecuri (Atanasoff 1925). While both microorganisms occur in Australia, they have not been seen in association with A. tritici in the field. In the early 1900s, A. tritici was found in wheat fields of the southern states of Australia and was considered a pest of concern. Now it is restricted to relatively few persistent populations in WAs northern wheatbelt, with its short growing season, dry summers and a proportion of fields with tight or continuous wheat rotations (Riley 1992; Fig. 3). However, over much of its former range, modern seed cleaning and crop rotations have made it locally extinct. This dramatic contraction of the range of A. tritici is also a consequence of the lack of naturalised hosts and little or no survival in the field through a growing season in the absence of a host. Agricultural impact In the early 1900s, the impact of A. tritici was on yield and quality of wheat, but there are no extant quantitative estimates of this impact. Currently, the impact is minor, but all wheat deliveries in WA are examined for A. tritici galls, and given that an average of 7.5 Mt of wheat is produced annually (1997--2007, ABARE), this is not without cost. Infested loads can be downgraded or rejected, and must be cleaned before redelivery to realise the full value of the grain. Several countries importing Australian wheat, barley and oats require area, crop or shipment freedom for A. tritici (PHYTO: plant and plant product export conditions database, Australian Quarantine and Inspection Service, http://www.aqis. gov.au/phyto, verified 21 February 2008). Control Under modern, mechanised agricultural practices, crop rotation and seed cleaning can provide full control of A. tritici. This is possible because (1) the galls are easily separated from grain for sowing, (2) the nematode has no naturalised hosts in Australia,

Australian anguinids

Australasian Plant Pathology

295

and (3) with apparently minimal carry-over to a second season in the absence of a host, a host-free break of 1 year is sufficient. The failure to achieve full control in the northern wheatbelt of WA might be, in part, a result of the short growing season and relatively dry soil surface allowing some survival into a second year in the absence of a host. It is somewhat ironic that the short in-field survival of A. tritici is in stark contrast to its unsurpassed longevity in an anhydrobiotic state (Limber 1973; Riley and Mirmoeini 2006). Other Australian anguinids In addition to the three species detailed above, the following anguinids are known from Australia: Anguina australis in Ehrharta longiflora (Steiner 1940; Riley et al. 2001a, 2001b), Anguina microlaenae in Microlaena stipoides (Fawcett 1938; de Silva and Riley 2002), Anguina sp. in Holcus lanatus (Riley et al. 1988), Mesoanguina mobilis in Arctotheca calendula (Chit and Fisher 1975), Subanguina radicicola in Lolium perenne and Poa spp. (McLeod et al. 1994), an undescribed seed-gall anguinid in Austrostipa scabra var. scabra (Riley et al. 1988) and an undescribed leaf gall anguinid in Astrebla pectinata (Powers et al. 2001). None of these have any direct economic impact. A. australis is common in WA but its host is a naturalised grass weed from South Africa for which the nematode provides little or no biological control. Anguina sp. in H. lanatus is widespread and associated with D. alopecuri infections but not R. toxicus, and again the host is only a weedy grass in pastures. Although S. radicicola (a root gall anguinid) is potentially a pest, it is not widespread and of no evident impact in Australia. Although the seed of the hosts that could be infested by some of these species are not normally traded, occasionally exporters of other seeds and grains are asked for a declaration of freedom from one or two of these species. One point for potential confusion is that Anguina agrostis sensu lato, which can include the Anguina sp. from H. lanatus, is considered a quarantine pest in some countries. However, molecular data clearly indicate that Anguina sp. from H. lanatus is a distinct species (Subbotin et al. 2004). Although not recorded in the field, some of the species may have the potential to be vectors for R. toxicus (Riley et al. 2001a). Discussion and conclusions Of the nematodes that affect plant and animal production, the plant gall-forming anguinids hold a unique place. Most plantparasitic nematodes damage roots, affect water and nutrient uptake, produce few, if any, specific aboveground symptoms and are largely spread with soil or vegetative propagating material. Most anguinids are very different. They survive and spread in seed or plant residues, produce specific aboveground symptoms, are highly adapted to the rigors of an exposed, aboveground life and are vectors for plant-colonising bacteria and fungi. This means they have direct effects on yield or quality of produce, are of greater quarantine concern and their association with a toxigenic bacterium gives them the unique status of affecting both plant and animal production. No nematode parasites of livestock are known to feed on plants, even though some have a free-living phase in pasture soil. In the Australian context, the anguinid vectors of R. toxicus, which have contributed to considerable economic and social

impact through ARGT and FPS, continue as an important and largely unique problem in world nematology. In the 1960s, a similar Anguina/Rathayibacter association in Festuca nigrescens in Oregon, in the United States, came and went before the causal agents and biology could be fully characterised. Over 100 refereed scientific papers on A. funesta, R. toxicus, corynetoxins and corynetoxin poisonings (ARGT and FPS) have been published, far more than can be given justice in this brief overview. This represents a considerable portion of the nematological research by South and Western Australian State government laboratories and an important component of plant toxicology research in the CSIRO over the last 30 years. The combined efforts of these researchers have provided a firm understanding of key aspects of this complex problem and to a large degree, provided workable solutions. However, there is much more to learn about Anguina/Rathayibacter associations, but with declining impacts, the perceived priority and financial support for further research on them in Australia has declined. Australias exotic and endemic anguinids are varied and intriguing but apart from the few with economic impact, they are not well studied. Future investigations have the potential to discover new species with unique and highly adapted biology. However, in the absence of economic imperatives, such work will remain for another generation of plant nematologists. Acknowledgements
Dr K. Davies is acknowledged for her encouragement and advice and Dr T. Bertozzi for helpful suggestions during preparation of the manuscript.

References
Anon. (1994) Annual ryegrass Lolium rigidum Guard. Plant Varieties Journal 7, 16. Anon. (2004) Hybrid short-lived ryegrass (Lolium hybrid) Safeguard. Plant Varieties Journal 17, 91. Anon. (2005) Standard for minimising risk of corynetoxin contamination of hay and straw for export. (Australian Quarantine and Inspection Service, Department of Agriculture Fisheries and Forestry: Barton, ACT) Atanasoff D (1925) The Dilophospora disease in cereals. Phytopathology 15, 11--40. Barbetti MJ, Riley IT (2006) Field application of Dilophospora alopecuri to manage annual ryegrass toxicity caused by Rathayibacter toxicus. Plant Disease 90, 229--232. doi: 10.1094/PD-90-0229 Bertozzi T (2003) Biology and control of the anguinid nematode associated with flood plain staggers. PhD Thesis, The University of Adelaide. Bertozzi T, McKay AC (1995) Incidence on Polypogon monspeliensis of Clavibacter toxicus and Anguina sp., the organisms associated with flood plain staggers in South Australia. Australian Journal of Experimental Research 35, 567--569. doi: 10.1071/EA9950567 Bird AF, McKay AC (1987) Adhesion of conidia of the fungus Dilophospora alopecuri to the cuticle of the nematode Anguina agrostis, the vector in annual ryegrass toxicity. International Journal for Parasitology 17, 1239--1247. doi: 10.1016/0020-7519(87)90088-9 Bird AF, Stynes BA (1977) The morphology of a Corynebacterium sp. parasitic on annual ryegrass. Phytopatholgy 67, 828--830. Bird AF, Stynes BA (1981a) The life cycle of Anguina agrostis: embryogenesis. International Journal for Parasitology 11, 23--33. doi: 10.1016/0020-7519(81)90022-9 Bird AF, Stynes BA (1981b) The life cycle of Anguina agrostis: postembryonic growth of the second stage larva. International Journal for Parasitology 11, 243--250. doi: 10.1016/0020-7519(81)90056-4

296

Australasian Plant Pathology

I. T. Riley and M. J. Barbetti

Bird AF, Stynes BA (1981c) The life cycle of Anguina agrostis: development in host plant. International Journal for Parasitology 11, 431--440. doi: 10.1016/0020-7519(81)90061-8 Bourke CA, Carrigan MJ, Love SCJ (1992) Flood plain staggers, a tunicaminyluracil toxicosis of cattle in northern New South Wales. Australian Veterinary Journal 69, 228--229. Brown AGP, Vogel P (1986) Annual ryegrass toxicity research update. Journal of Agriculture Western Australia 27, 3--6. [4th series] Bryden WL, Trengrove CL, Davis EO, Giesecke PR, Curran GC (1994) Corynetoxicosis of livestock: a nematode-bacterium disease complex associated with different grasses. In Plant-associated toxins: agricultural, phytochemical and ecological aspects. (Eds SM Colegate, PR Dorling) pp. 410--415. (CAB International: Wallingford, UK) Burdass WJ (1986) Control strategies for annual ryegrass toxicity. Journal of Agriculture Western Australia 27, 710. [4th series] Chit W, Fisher JM (1975) Anguina mobilis n. sp., a parasite of capeweed (Arctotheca calendula). Nematologica 21, 53--61. Chizhov VN, Subbotin SA (1987) Revision of the nematode subfamily Anguininae (Nematoda: Tylenchida) on the basis of their biological characteristic. In English translations of selected taxonomic papers in nematology. Vol. 4. (Ed. R Fortuner) pp. 5--18. (California Department of Food and Agriculture: Sacramento, CA) Clarke R, Nurse G (1999) Annual ryegrass toxicity. Agricultural Notes AG0129, Department of Primary Industries, Victoria. 3 pp. Davies SC, Williams IH, White CL, Allen JG, Edgar JA, Cockrum PA, Stewart P (1993) Protective effect of cobalt against apparent liver damage in sheep exposed to toxic annual ryegrass. Australian Veterinary Journal 70, 186--187. Davies SC, White CL, Williams IH, Allen JG, Croker KP (1996) Sublethal exposure to corynetoxins affects production of grazing sheep. Australian Journal of Experimental Agriculture 36, 649--655. doi: 10.1071/ EA9960649 Davis EO, Curran GE, Hetherington WT, Norris DA, Wise GA, Roth IJ, Seawright AA, Bryden WL (1995) Clinical, pathological and epidemiological aspects of flood plain staggers, a corynetoxicosis of livestock grazing Agrostis avenacea. Australian Veterinary Journal 72, 187--190. de Silva P, Riley IT (2002) Aspects of the survival and reproduction of Anguina microlaenae (Nematoda: Anguinidae). Transactions of the Royal Society of South Australia 126, 45--49. Edgar J (2004) Future impact of food safety issues on animal production and trade: implication for research. Australian Journal of Experimental Agriculture 44, 1073--1078. doi: 10.1071/EA03222 Edgar JA (1994) Toxins in temperate grasses -- implications and solutions. New Zealand Journal of Agricultural Research 37, 341--347. Edgar JA, Fralm JL, Cockrum PA, Anderton N, Jago MV, Culvenor CCJ, Jones AJ, Murray K, Shaw KJ (1982) Corynetoxins, causative agents of annual ryegrass toxicity; their identification as tunicamycin group antibiotics. Journal of the Chemical Society, Chemical Communications, 222--224. doi: 10.1039/c39820000222 Edgar JA, Cockrum PA, Stewart PL, Anderton NA, Payne AL (1994) Identification of corynetoxins as the cause of poisoning associated with annual beardgrass [Polypogon monspeliensis (L.) Desf.] and blown grass (Agrostis avenacea C. Gemelin). In Plant-associated toxins: agricultural, phytochemical and ecological aspects. (Eds SM Colegate, PR Dorling) pp. 393--398. (CAB International: Wallingford, UK) Fawcett SGM (1938) A disease of the Australian grass Microlaena stipoides R. Br. caused by a nematode Anguillulina microlaenae n. sp. Journal of Helminthology 16, 17--32. Finnie JW (2006) Review of corynetoxins poisoning of livestock, a neurological disorder produced by a nematode-bacterium complex. Australian Veterinary Journal 84, 271--277. doi: 10.1111/j.17510813.2006.00019.x

Gibson IAS, Sutton BC (1976) Dilophospora alopecuri. Commonwealth Mycological Institute Descriptions of Pathogenic Fungi and Bacteria No. 490. (Commonwealth Mycological Institute: Kew, UK) Gill GS (1996a) Why annual ryegrass is a problem in Australian agriculture. Plant Protection Quarterly 11(Suppl. 1), 193--195. Gill GS (1996b) Ecology of annual ryegrass. Plant Protection Quarterly 11 (Suppl. 1), 195--198. Gwyn R, Hadlow AJ (1971) Toxicity syndrome in sheep grazing Wimmera ryegrass in Western Australia. Australian Veterinary Journal 47, 408. Jago MV, Payne AL, Peterson JE, Bagust TJ (1983) Inhibition of glycosylation by corynetoxin, the causative agent of annual ryegrass toxicity: a comparison with tunicamycin. Chemical-Biological Interactions 45, 223--234. doi: 10.1016/0009-2797(83)90070-4 Krall EL (1991) Wheat and grass nematodes: Anguina, Subanguina, and related genera. In Manual of agricultural nematology. (Ed. WR Nickel) pp. 721--760. (Marcel Dekker: New York) Lee C, Colegate S, Fisher AD (2006) Development of a maze test and its application to assess spatial learning and memory in Merino sheep. Applied Animal Behaviour Science 96, 43--51. doi: 10.1016/j. applanim.2005.06.001 Limber DP (1973) Notes on the longevity of Anguina tritici (Steinbuch, 1799) Filipjev, 1936, and its ability to invade wheat seedlings after thirty-two years of dormancy. Proceedings of the Helminthological Society of Washington 40, 272--274. Maling I, OCallaghan AM (1992) Research, farm monitoring and extension of ARGT control using herbicide/grazing strategies. Final Report on Wool Research Corporation project DAW 71P. Western Australian Department of Agriculture Miscellaneous Publication No. 12/92. 59 pp. Masters AM, Gregory AR, Evans RJ, Speijers JE, Sutherland SS (2006) An enzyme-linked immunosorbent assay for the detection of Rathayibacter toxicus, the bacterium involved in annual ryegrass toxicity, in hay. Australian Journal of Agricultural Research 57, 731--742. doi: 10.1071/AR03125 Matthews JM (1996a) Cultural management of annual ryegrass. Plant Protection Quarterly 11(Suppl. 1), 198--200. Matthews JM (1996b) Chemical management of annual ryegrass. Plant Protection Quarterly 11(Suppl. 1), 200--202. May C, Stewart PL (1998) Development of a toxin-binding agent as a treatment for tunicaminyluracil toxicity: protection against tunicamycin poisoning of sheep. Australian Veterinary Journal 76, 752--756. McIntosh GH, Rac R, Thomas MR (1967) Toxicity of parasitised Wimmera ryegrass, Lolium rigidum, for sheep and cattle. Australian Veterinary Journal 43, 349--353. McKay AC (1985) Investigations to develop methods to control the nematode associated with annual ryegrass toxicity. PhD Thesis, The University of Adelaide, Australia. McKay AC (1993) Development of annual ryegrass resistant to Anguina funesta, the vector in annual ryegrass toxicity. In Pests of pastures: weed, invertebrate and disease pests of Australian sheep pastures. (Ed.ESDelfosse)pp.226--229.(CSIROInformationServices:Melbourne) McKay AC, Ophel KM (1993) Toxigenic Clavibacter/Anguina associations infecting grass seedheads. Annual Review of Phytopathology 31, 153--169. doi: 10.1146/annurev.phyto.31.1.153 McKay AC, Riley IT (1993) Sampling ryegrass to assess the risk of annual ryegrass toxicity. Australian Veterinary Journal 70, 241--243. AJ (1981) Ecological field studies on Anguina McKay AC, Fisher JM, Dube funesta, the vector in annual ryegrass toxicity. Australian Journal of Agricultural Research 32, 917--927. doi: 10.1071/AR9810917 McKay AC, Fisher JM, Giesecke R, Crosby J (1985) Are farmers controlling annual ryegrass toxicity in South Australia? In Plant toxicology. Proceedings of the Australia-USA poisonous plants symposium. (Eds AA Seawright, MP Hegarty, LF James, RF Keeler) pp. 559--568. (Queensland Poisonous Plants Committee, Queensland Department of Primary Industries, Animal Research Institute: Yeerongpilly)

Australian anguinids

Australasian Plant Pathology

297

McKay AC, Ophel KM, Reardon TB, Gooden JM (1993) Livestock deaths associated with Clavibacter toxicus/Anguina sp., infection in seedheads of Agrostis avenacea and Polypogon monspeliensis. Plant Disease 77, 635--641. McLeod R, Reay F, Smyth J (1994) Plant nematodes of Australia listed by plant and by genes. (NSW Agriculture: Orange) Nogawa M, Ishikawa T, Miyazaki S, Suto Y, Sato W, Taneichi A, Kobayashi M (1997) Ryegrass intoxication in cattle and sheep fed oaten hay imported from Australia. Journal of the Japan Veterinary Medical Association 50, 321--326. Powers TO, Szalanski AL, Mullin PG, Harris TS, Bertozzi T, Griesbach JA (2001) Identification of seed gall nematodes of agronomic and regulatory concern with PCR-RFLP of ITS1. Journal of Nematology 33, 191--194. Preston CM, Bird AF (1987) Physiological and morphological changes associated with recovery from anabiosis in the dauer larva of the nematode Anguina agrostis. Parasitology 95, 125--133. Price PC, Fisher JM, Kerr A (1979) Annual ryegrass toxicity: parasitism of Lolium rigidum by a seed gall forming nematode (Anguina sp.). Annals of Applied Biology 91, 359--369. Retallick MAS, Colegate SM, Michalski WP (2007) A toxicoproteomic approach to understanding annual ryegrass toxicity. In Poisonous plants: global research and solutions. (Eds KE Panter, TL Wierenga, JA Pfister) pp. 230--234. (CAB International: Wallingford, UK) Riley IT (1992) Anguina tritici is a potential vector of Clavibacter toxicus. Australasian Plant Pathology 21, 147--148. doi: 10.1071/APP9920147 Riley IT (1994a) Dilophospora alopecuri and decline in annual ryegrass toxicity in Western Australia. Australian Journal of Agricultural Research 45, 841--850. doi: 10.1071/AR9940841 Riley IT (1994b) Opportunities for biological control of the toxigenic Anguina/Clavibacter association in annual ryegrass. In Plantassociated toxins: agricultural, phytochemical and ecological aspects. (Eds SM Colegate, PR Dorling) pp. 439--444. (CAB International: Wallingford, UK) Riley IT (1995) Vulpia myuros and the annual ryegrass toxicity organisms, Anguina funesta, and Clavibacter toxicus. Fundamental and Applied Nematology 18, 595--598. Riley IT (1996) Dilophospora alopecuri on Lolium rigidum and Holcus lanatus in south-eastern Australia. Australasian Plant Pathology 25, 255--259. doi: 10.1071/AP96047 Riley IT, Bertozzi T (2004) Variation in sex ratios in four Anguina (Nematoda: Anguinidae) species. Transaction of the Royal Society of South Australia 128, 43--46. Riley IT, Gill GS (1994) Relationship between herbicide resistance in Lolium rigidum and populations of Anguina funesta, the nematode vector in annual ryegrass toxicity. Australian Journal of Experimental Agriculture 34, 633--635. doi: 10.1071/EA9940633 Riley IT, McKay AC (1990) Specificity of the adhesion of some plant pathogenic micro-organisms to the cuticle of nematodes in the genus Anguina (Nematoda: Anguinidae). Nematologica 36, 90--103. Riley IT, McKay AC (1991a) Invasion of some grasses by Anguina funesta (Nematoda: Anguinidae) juveniles. Nematologica 37, 447--454. Riley IT, McKay AC (1991b) Susceptibility of Anguina funesta (Nematoda: Anguinidae) populations to adhesion by the toxigenic Clavibacter sp. responsible for annual ryegrass toxicity. Nematologica 37, 439--446. Riley IT, Mirmoeini Z (2006) Longevity of Anguina tritici. Australasian Nematology Newsletter 17, 16. Riley IT, Reardon TB (1995) Isolation and characterisation of Clavibacter tritici associated with Anguina tritici in Western Australia. Plant Pathology 44, 805--810. doi: 10.1111/j.1365-3059.1995.tb02739.x

Riley IT, Yan G (1998) Evaluation of non-toxigenic, nematode-vectored Clavibacter species for potential control of Clavibacter toxicus, the bacterium responsible for annual ryegrass toxicity. 1998 Annual Scientific Meeting of the Australian Society of Microbiology, 27 September--2 October 1998, Hobart, Tasmania. Microbiology Australia 19(4), A124. Riley IT, Reardon TB, McKay AC (1988) Electrophoretic resolution of species boundaries in seed-gall nematodes, Anguina spp. (Nematoda: Anguinidae), from some graminaceous hosts in Australia and New Zealand. Nematologica 34, 401--411. Riley IT, Schmitz A, de Silva P (2001a) Anguina australis, a vector for Rathayibacter toxicus in Ehrharta longiflora. Australasian Plant Pathology 30, 361--364. doi: 10.1071/AP01048 Riley IT, Shedley D, Sivasithamparam K (2001b) Anhydrobiosis and reproduction in Anguina australis. Australasian Plant Pathology 30, 361--364. doi: 10.1071/AP01048 Roberts WD, Mlodawski G, Macdonagh A, Gibson R, Bucat J (1994) The distribution of annual ryegrass toxicity in Western Australia. In Plantassociated toxins: agricultural, phytochemical and ecological aspects. (Eds SM Colegate, PR Dorling) pp. 51--56. (CAB International: Wallingford, UK) Southey JF (1972) Anguina tritici. CIH description of plant parasitic nematodes. Set 1, No. 13. (Commonwealth Institute of Helminthology: St Albans, UK) Steiner G (1940) Opuscula miscellanea nematologica. VIII. Proceedings of the Helminthological Society of Washington 7, 54--62. Subbotin SA, Krall EL, Riley IT, Chizhov VN, Staelens A, De Loose M, Moens M (2004) Evolution of the gall-forming plant parasitic nematodes (Tylenchida: Anguinidae) and their relationships with hosts as inferred from internal transcribed spacer sequences of nuclear ribosomal DNA. Molecular Phylogenetics and Evolution 30, 226--235. doi: 10.1016/ S1055-7903(03)00188-X Swarup G, Gupta P (1971) On the ear-cockle and tundu diseases of wheat. II. Studies on Anguina tritici and Corynebacterium tritici. Indian Phytopathology 24, 359--365. Than KA, Cao Y, Michalewicz A, Edgar JA (1998a) Development of an immunoassay for corynetoxins. In Toxic plants and other natural toxicants. (Eds T Garland, AC Barr) pp. 49--54. (CAB International: Wallingford, UK) Than KA, Cao Y, Michalewicz A, Edgar JA (1998b) Development of a vaccine against annual ryegrass toxicity. In Toxic plants and other natural toxicants. (Eds T Garland, AC Barr) pp. 165--168. (CAB International: Wallingford, UK) Yan G, Riley IT (1998) Using twist fungus (Dilophospora alopecuri) to reduce the risk of annual ryegrass toxicity. Agriculture Western Australia Farmnote No. 33/98. 4 pp. Yan G, Riley IT (2003) Fermentation period, moisture content, and substrate effects on inoculum effectiveness of Dilophospora alopecuri: a biocontrol agent of annual ryegrass toxicity. Biological Control 26, 174--179. doi: 10.1016/S1049-9644(02)00131-7 Yan G, Riley IT (2005) Variation in pathogenicity between isolates of Dilophospora alopecuri, an antagonist of the causal organisms of annual ryegrass toxicity. Australian Journal of Experimental Agriculture 45, 1157--1162. doi: 10.1071/EA04012

Manuscript received 20 January 2008, accepted 11 February 2008

http://www.publish.csiro.au/journals/app

Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.