Use of Green Fluorescent Protein-Transgenic Strains to Study Pathogenic and Nonpathogenic Lifestyles in Colletotrichum acutatum

ABSTRACT Colletotrichum acutatum, which causes anthracnose disease on strawberry, can also persist on several other plant species without causing disease symptoms. The genetic and molecular bases that determine pathogenic and nonpathogenic lifestyles in C. acutatum are unclear. We developed a transformation system for C. acutatum by electroporation of germinating conidia, and transgenic isolates that express the green fluorescent protein (GFP) were produced. Details of the pathogenic and nonpathogenic lifestyles of C. acutatum were determined by using GFP-transgenic isolates. Major differences between colonization-mediating processes of strawberry and of other plants were observed. On the main host, strawberry, the germinating conidia formed branched, thick hyphae, and large numbers of appressoria were produced that were essential for plant penetration. In strawberry, the fungus developed rapidly, filling the mesophyll with dense mycelium that invaded the cells and caused necrosis of the tissue. In nonpathogenic interactions on pepper, eggplant, and tomato, the conidia germinated, producing thin, straight germ tubes. Appressoria were produced but failed to germinate and penetrate leaf tissue, resulting in epiphytic growth without invasion of the plant. Penetration of the plant occurred only several days after inoculation and was restricted to the intercellular spaces of the first cell layers of infected tissue without causing any visible damage. Much of the new fungal biomass continued to develop on the surface of inoculated organs in the nonpathogenic interaction. The differences in fungal development on strawberry compared with the other plant species suggest that signal molecules, which may be present only in strawberry, trigger appressorial germination and penetration of the primary host.     

Dynamics of insecticide resistance in Bemisia tabaci: a case study with the insect growth regulator pyriproxyfen

The dynamics of pyriproxyfen resistance in Bemisia tabaci (Gennadius) have been studied intensively in cotton fields and greenhouses in Israel. High resistance to pyriproxyfen evolved one year after its introduction for use on flowers in greenhouses, after three successive applications. After ten years of pyriproxyfen use in cotton fields, with only one application per season, a high level of resistance was observed, but its rate of development differed among localities. It is hypothesized that these differences reflect temporal and spatial variations in the availability of alternative host plants for B tabaci, and that pyriproxyfen resistance evolved most readily in geographically isolated areas. It is also likely that the occurrence and development of resistance is partially biotype-related. Pyriproxyfen use has ceased in high-resistance localities, but in areas with low frequencies of resistance, cotton growers have continued to achieve acceptable control of B tabaci with pyriproxyfen. Due to the absence of applications of pyriproxyfen in some cotton fields, resistance levels tended to decline between 1998 and 2001. Laboratory experiments support the hypothesis that this decline reflects, in part, fitness costs associated with pyriproxyfen resistance.     

Note: Effect of the surfactant BB5 on the potency of thiamethoxam against the whiteflyBemisia tabaci

BB5, an alkyl phenoxy polyethylene ethanol formulated in acidic buffer, is an adjuvant added usually to various pesticides to potentiate their activities on various biological systems. Addition of 0.2% BB5 resulted in a considerable increase of thiamethoxam toxicity onBemisia tabaci (Gennadius) nymphs. According to LC₅₀ values, the combined effect was over 25-fold higher than that of thiamethoxam alone. On the other hand, no appreciable combined effect was observed when BB5 and thiamethoxam were applied together onB. tabaci adults. The importance of these results is discussed. http://www.phytoparasitica.org posting Dec. 20, 2004.     

Toxicity of spiromesifen to the developmental stages of Bemisia tabaci biotype B

BACKGROUND: Spiromesifen is a novel insecticidal/acaricidal compound derived from spirocyclic tetronic acids that acts effectively against whiteflies and mites via inhibition of acetyl‐CoA‐carboxylase, a lipid metabolism enzyme. The effects of spiromesifen on the developmental stages of the whitefly Bemisia tabaci (Gennadius) were studied under laboratory conditions to generate baseline action thresholds for field evaluations of the compound. RESULTS: Adult B. tabaci mortality rate after spiromesifen treatment (5 mg L^?1) was 40%. Treatment with 0.5 mg L^?1 reduced fecundity per female by more than 80%, and fertility was almost nil. LC₅₀ for eggs was 2.6 mg L^?1, and for first instar 0.5 mg L^?1. Scanning electron microscopy revealed that eggs laid by treated adult females had an abnormally perforated chorion, and females were unable to complete oviposition. Light and fluorescent microscopy showed significantly smaller eggs following treatment, and smaller, abnormally formed and improperly localized bacteriomes in eggs and nymphs. The number of ovarioles counted in females treated with 5 mg L^?1 was significantly reduced. Spiromesifen showed no cross‐resistance with other commonly used insecticides from different chemical groups, and resistance monitoring in Israel showed no development of field resistance to this insecticide after 1 year of use. CONCLUSION: The strong effect on juvenile stages of B. tabaci with a unique mode of action and the absence of cross‐resistance with major commonly used insecticides from different chemical groups suggest the use of spiromesifen in pest and resistance management programmes. Copyright © 2008 Society of Chemical Industry     

Effect of milbemectin on the sweetpotato whitefly,Bemisia tabad

Milbemectin has a chemical structure close to the group of avermectins, which are derived fromStreptomyces avermitilis, and is considered primarily an efficient miticide. Effects of milbemectin on the sweetpotato whitefly,Bemisia tabaci Gennadius, were investigated under laboratory and field conditions. In bioassays conducted under controlled chamber conditions, the compound affected 1st instars ofB. tabaci, resulting in a LC₉₀ of 0.06 mg a.i. I-1. Later stage larvae were much less affected. Milbemectin is highly photodegradable in sunlight. In laboratory assays, when treated cotton seedlings were subjected to 3 h of sunlight before being exposed toB. tabaci adults, no mortality of the whiteflies was observed. Milbemectin at a concentration of 2 mg a.i. I-1 applied in combination with 0.2% ‘Ultra Fine’ mineral oil showed a residual activity of 67% adult mortality 10 days after application, whereas milbemectin alone had no appreciable activity. The effect of milbemectin on whitefly populations in a cotton field was compared with that of cypermethrin and of untreated control. Although milbemectin was not applied with mineral oil, it was more effective than cypermethrin in controlling the whitefly populations. This insecticide/miticide seems not to affect appreciably natural enemies ofB. tabaci. Milbemectin may be considered a compound with the potential for controllingB. tabaci populations. Mineral oils enhanced the potency of milbemectin on both whitefly larvae and adults. http://www.phytoparasitica.org posting July 27, 1999. Contribution No. 501/99 from the Inst. of Plant Protection, Agricultural Research Organization.     

Potassium salt microinjection into Xenopus oocytes mimics gonadotropin treatment

Gonadotropin stimulates protein synthesis and growth in ovarian oocytes. The hormone is also known to modify transfollicular K+ fluxes and is now shown to cause increased intraoocytic K+ activity (aK). The hormone's effect on aK was duplicated by microinjecting K+ salts into oocytes which were incubated in paraffin oil. This treatment mimicked the influence of gonadotropin on both the rate of protein synthesis and the synthesis of specific polypeptides. These findings suggest that gonadotropin-stimulated oocyte growth is attributable largely to the hormone's influence on transfollicular K+ fluxes. They support the hypothesis that the K+ flux and aK changes observed during cell activation are critical in causing subsequent increases in protein synthesis and growth.     

Stratigraphic sections, bedding sequences, and random processes

Both dependent and independent random processes can be used to study and synthesize stratigraphic sections and bedding sequences. Synthetic stratigraphic sections can be modeled to fit any geologic problem with a good correspondence between the synthetic and actual stratigraphic sections. To synthesize a sedimentary sequence, one needs only a transition procedure to go from one lithology or bedding type to another, and frequency distributions of thickness for the various lithologies. A sedimentation process with a "memory" is termed Markovian; if the past has no influence on either present or future sedimentation, it is called an independent trials process.