Distribution in time and space of resistance to the pathogenic fungus Erynia neoaphidis in the pea aphid Acyrthosiphon pisum

Two biotypes of pea aphid, Acyrthosiphon pisum, have been recognized in Australia, by their susceptibility (S) or resistance (R) to certain isolates of the fungal pathogen Erynia neoaphidis. The responses of these two biotypes to 40 isolates of the fungus have shown that the R form is largely confined to two States, New South Wales and Victoria, but appears to have recently spread into Queensland and Tasmania. There is no evidence to suggest it occurs outside Australia. Sequential sampling of two field populations of pea aphids during 1981 and 1982 showed that the proportion of R form remained stable at 10.7±3.0 and 14.6±2.6% (mean±standard error) for the two populations. Glasshouse competition experiments run at the comparatively high temperature of 25°C resulted in the R form becoming dominant even when the initial ratio was 4:1 in favour of the S form. The ecological and genetical implications are discussed.

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Résumé Deux types biologiques d'Acyrthosiphon pisum ont été définis, en Australie, suivant leur sensibilité (S) ou leur résistance (R) à certains isolats du champignon pathogène Erynia neoaphidis. Les réponses des deux biotypes à 40 isolats du champignon ont montré que la forme R est essentiellement cantonnée à deux états: Nouvelle Galle due Sud et Victoria, mais a récemment gagné le Queensland et la Tasmanie. Aucun élément ne fait dire qu'elle existe hors d'Australie. Des échantillonnages séquentiels des deux populations de pucerons dans la nature en 1981 et 1982 ont montré que la proportion de la forme R est restée stable à 10,7 et 14,62 dans les deux populations. Des expériences de compétition en serre à la température relativement haute de 25°C ont rendu la forme R dominante, même quand le rapport initial était de 4/1 en faveur de la forme S. La discussion porte sur les conséquences écologiques et génétiques.

     

A novel computerised image analysis method for the measurement of production of conidia from the aphid pathogenic fungus Erynia neoaphidis

A semi-automated method has been developed for the quantification and measurement of conidia discharged by the aphid pathogen Erynia neoaphidis. This was used to compare conidiation by E. neoaphidis-mycosed pea aphid cadavers, mycelial plugs cut from agar plates, mycelial pellets from shake flasks and by mycelial pellets from different phases of liquid batch fermenter culture. Aphid cadavers discharged significantly more and significantly smaller conidia than plugs or pellets. The volume of conidia discharged was stable over the period of discharge (80 h), but more detailed analysis of the size frequency distribution showed that more very small and very large conidia were discharged after 5 h incubation than after 75 h incubation. Biomass harvested at the end of the exponential growth phase in batch fermenter culture produced significantly more conidia than biomass from any other growth phase. The implications of these findings for the development of production and formulation processes for E. neoaphidis as a biological control agent are discussed.     

Ultrastructure and behaviour of the spindle pole body of the aphid-pathogenic fungusErynia neoaphidis

Summary A detailed account of the ultrastructure and behaviour of the spindle pole body (SPB) of the entomophthoraceous fungusErynia neoaphidis is presented for the first time.The SPB consists of extranuclear (ENC) and intranuclear (INC) components. The ENC is a saucepan-shaped structure which lies in a pocket of the nuclear envelope. It is composed of a forked, fibrillar handle and a shallow, cylindrical pan. The pan has a wall of two layers, both of which are thickened with a regular periodicity so that they appear to be beaded. It is postulated that the pan is formed from rough endoplasmic reticulum and that it synthesizes the amorphous, electron-dense material coating the ENC.The INC is a saucer-shaped, electron-dense plaque in which the ends of the spindle microtubules terminate. During metaphase, a clear zone separates the INC from the nuclear envelope and persists until telophase. The roles of the amorphous, electron-dense material and the clear zone as well as the method of SPB replication are discussed.     

Effect of oleic acid on vegetative growth of the aphid-pathogenic fungus Erynia neoaphidis

Abstract The aphid-pathogenic fungus Erynia neophidis grew on a semi-defined medium containing 16 g·1⁻¹ glucose, 3 g·1⁻¹ yeast extract and 5 g·1⁻ mycological peptone only when the medium was supplemented with low concentrations of certain fatty acids. Of these, oleic acid fulfilled the growth requirement at a concentration of 0.02% (v/v), but higher concentrations were toxic, causing complete loss of viability of cultures at a concentration of 0.2% (v/v) in liquid medium and at 4% (v/v) on solid medium. The reduced viability of the fungus in liquid culture compared to that on equivalent solid medium, and at low inoculum density compared to high inoculum density in liquid medium, is explicable in terms of this toxicity.     

Targeted Dispersal of the Aphid Pathogenic Fungus Erynia neoaphidis by the Aphid Predator Coccinella septempunctata

The potential of adult and larval C. septempunctata to vector the aphid-specific entomopathogenic fungus E. neoaphidis was assessed through a series of laboratory and field experiments. The ability of coccinellids to vector conidia from a colony of E. neoaphidis -infected pea aphids, Acyrthosiphon pisum, to a colony of uninfected A. pisum was demonstrated in a laboratory study. Adult coccinellids which had previously foraged on plants infested with different densities of sporulating cadavers (1, 5, 15, 30 cadavers per plant) initiated infection in a proportion of uninfected pea aphids (4, 0, 2 and 8%, respectively) when subsequently allowed to forage on A. pisum infested bean plants. Further laboratory studies demonstrated that fourth instar larvae and adult coccinellids artificially inoculated with conidia initiated infection in 11 and 13% of an A. pisum population in which they foraged, respectively. Furthermore, a proportion of A. pisum placed on bean plants which had previously been foraged on by inoculated larval and adult coccinellids also died from infection (3 and 10% of A. pisum, respectively). However, although coccinellid adults inoculated with conidia initiated infection in 19% of A. pisum, cereal aphids, S. avenae , exposed to the inoculated coccinellids did not become infected. A further laboratory study demonstrated that infection of A. pisum only occurred if inoculated coccinellids were transferred to A. pisum populations immediately post inoculation. However, a proportion of A. pisum placed on bean plants which had been foraged on by inoculated coccinellids transferred 0, 4 and 24 h post inoculation died from infection (9, 3 and 7%, respectively). A field study further demonstrated the potential of coccinellids to vector E. neoaphidis. Single spring sown field bean plants (Long Hoos Experimental Plots, IACRRothamsted Farm) were enclosed within nylon mesh bags and 25 adult A. pisum were added to each bag with one of the following treatments: no further addition (control), coccinellid adult (control), inoculated coccinellid adult, inoculated A. pisum or sporulating A. pisum cadavers. No aphids died of E. neoaphidis in the control treatments; 5, 16 and 33% of aphids were infected with E. neoaphidis on the other treatments, respectively.     

Influence of Flight and Colour Morph on Susceptibility of Sitobion avenae to Infection by Erynia neoaphidis

The influence of clone and morph on the infection of Sitobion avenae with Erynia neoaphidis was examined in laboratory assays. The LC 50 -values were low; green alates: 0.8-1.8, green apterae: 2.9, brown alates: 1.5 and brown apterae: 2.6-3.6 conidia mm -2 . Within each of the two inoculation occasions, alates of both clones were significantly more susceptible than apterae, while there was no difference between the two clones. The LT 50 -values at 18°C varied between 6.4 and 7.5 days and there was no significant difference in LT 50 -values between either the two different coloured clones or between alates and apterae.     

Influence of commercially derived lipids and a surfactant on the mode of germination and process of germ-tube formation in primary conidia of two species of Erynia subgenus Neopandora (Zygomycotina: Entomophthorales

Primary conidia of the entomopathogens Erynia (subgenus Neopandora) delphacis (1 isolate) and Erynia ( Neopandora) neoaphidis (3 isolates) were stimulated to form germ-tubes with Tween 20 and with free, long-chain fatty acids, each incorporated into Entomophthora complete medium (ECM). When combined with other basal media (three tested), these compounds did not stimulate germ-tube formation. Triacylglycerols and vegetable oils, added to the same media, allowed almost complete resporulation in the fungi. In both species, Tween 20 (0.1%) encouraged greater germ-tube production (41–69%) than the fatty acids (0.1%) (≤36%). For E. delphacis, Tween 20 and the fatty acids differed significantly, but for E. neoaphidis the differences were almost always insignificant. Myristic and oleic acids stimulated germ-tube formation in both species. Palmitic acid allowed almost complete resporulation of the fungi, except for one isolate of E. neoaphidis that formed germ-tubes. Linoleic acid, tested only for E. delphacis, was fungistatic to most conidia. Higher concentrations of the fatty acids (≤1%) did not increase germ-tube formation, except 1% oleic acid which affected E. delphacis alone (>80% germination and germ-tubes). Linoleic acid, and sometimes also myristic and oleic, were fungistatic and/or toxic, depending on their concentration and on medium composition. Addition of fatty acids to ECM usually extended the lag period, and altered the morphology of the conidia and germ-tubes. These phenomena were not observed with Tween 20. Colonies were formed by E. delphacis alone, stimulated by ECM supplemented with Tween 20 or fatty acids. The results are discussed with respect to biological and physiological aspects of germination, and with respect to the mode of action of the fatty acids and the surfactant. This revised version was published online in August 2006 with corrections to the Cover Date.     

虫霉目新种和新记录

本文报道了虫霉目的一个新种和3个新记录种。新种叶蝉虫疫霉(Erynia cicadellisLi et Fan sp.nov.)是我国南方稻叶蝉虫霉流行病的病原之一,与近缘种飞虱虫疫霉[E．Deiphaeis (Heri) Humber et Ben-Ze’ev] 的主要区别在于后者孢子较大,且不具有假根。蚜虫疫霉[E. aphidis (Hoffm.) Humber et Ben-Ze'ev]系世界不常见种,国内首次在陕西岚皋及福州的桃蚜上记录;飞虱虫疫霉和佩氏虫疫霉[E．Petchii Ben-Ze'ev et Kenneth)皆系南方褐稻虱虫霉流行病的病原。     

飞虱虫疫霉的产孢生物学研究

飞虱虫疫霉Erynia delphacis (Hori) 是水稻害虫稻褐飞虱的天敌,在田间常造成流行病,使大量褐飞虱死亡。在研究利用真菌防治害虫方面,一个很重要的问题就是如何在人工培养基上培养病原菌,并获得大量可以保存的孢子然后用于田间。本试验的重点在于用液体培养的方法获得这种菌的分生孢子,并测定了液体和固体培养的产孢量和产孢时间。