Repression of deoxynivalenol accumulation and expression of Tri genes in Fusarium culmorum by fungicides in vitro

A defined medium was developed in which to monitor deoxynivalenol (DON) accumulation, fungal growth and expression of genes involved in trichothecene biosynthesis (designated Tri genes). In liquid culture, DON accumulated shortly after maximum expression of Tri6 and coincident with expression of Tri5. This was generally 96 h after inoculation. The effects of sublethal concentrations of the fungicides azoxystrobin, trifloxystrobin, kresoxim-methyl and tebuconazole on biosynthesis of the trichothecene DON by Fusarium culmorum were studied using this medium. The strobilurin fungicides trifloxystrobin and azoxystrobin significantly reduced the accumulation of DON in culture medium at a range of concentrations. Kresoxim-methyl, also a strobilurin, and tebuconazole, a triazole, did not significantly reduce the accumulation of DON, although levels were lower than those in nonamended cultures. Trifloxystrobin significantly reduced the accumulation of DON when added to cultures before initiation of trichothecene biosynthesis. RT-PCR assays of the expression of Tri6 and Tri5 genes indicated that trifloxystrobin acted by inhibiting the initiation of trichothecene biosynthesis.     

Factors affecting seed dormancy and germination of Johnsongrass, Sorghum halepense (L.) Pers.

Seeds of Johnsongrass [Sorghum halepense (L.) Pers.] germinated to higher percentages (20–30% higher) when incubated at 28 and 35° C than at 10 or 22° C. After-ripening was accelerated by dry storage of these seeds at 50°C. Seeds pre-chilled at 6°C for 2–4 weeks followed by incubation at 28°C germinated 40–60%. Light effects on germination were related to incubation temperatures; inhibitory at 22°C; no response at 28°C; and stimulatory at 35°C. Effects of gibberellin A₃ (GA₃) also varied depending on incubation temperature, sodium hypochlorite (NaOCl) immersion and light conditions. Immersion of dry seeds in either 700 mM NaOCl, 900 mM H₂O₂ or concentrated H₂SO₄ before incubation in water was effective in breaking dormancy. This result suggests the modes of action of H₂SO₄ in the termination of dormancy may be similar to those of NaOCl and H₂SO₄ as previously suggested by Hsiao & Quick (1984), that is by modification or scarification of the hull or seed coat membranes, and also by the supply of additional oxygen to the seed.     

The joint action of fungicides in mixtures: comparison of two methods for synergy calculation

Two methods for evaluating the joint action of fungicides in mixtures were analysed. In order to obtain a relatively rapid answer on the type of interaction between fungicides in a mixture (additivity, synergism or antagonism), one can apply the Abbott formula to data on fungus survival. Tests with this method will not be accurate at high effective dose values. A more accurate determination of the joint action of fungicides can be made by the Wadley method, applied to data on effective doses. This involves more experimental work than the Abbott method. Optimization of mixing ratios of fungicides required a set of experimental data on effective doses with several mixing ratios.     

Identification of divergent variants of Grapevine virus A

Eight isolates of Grapevine virus A (GVA), which induced different symptoms in leaves of Nicotiana benthamiana, were recovered from various grapevines. The dsRNA patterns of two isolates, which consistently induced mild vein clearing (referred here as mild isolates of GVA) were similar, but different from those of other isolates of GVA. Analysis based on overall nucleotide (nt) sequence identity in the 3 terminal part of the GVA genome, comprising part of ORF3 (putative movement protein, MP), entire ORF4 (capsid protein, CP), entire ORF5 and part of 3 UTR, revealed that GVA isolates separate into three groups (I, II, III), sharing 91.0–99.8% nt sequence identity within groups and 78.0–89.3% nt sequence identity between groups. Mild isolates of the virus were group III and shared only 78.0–79.6% nt sequence identity with the other isolates. The comparison of predicted amino acid sequences for MP and CP revealed many amino acid alterations, revealing distinct local net charges of these proteins for mild isolates of the virus. Based on both conserved and divergent nt regions in the CP and ORF5, oligonucleotide primers were designed for the simultaneous RT-PCR detection of all GVA isolates and for the specific detection of the most divergent virus variants represented here by mild isolates of the virus.     

单克隆抗体检测栗疫菌dsRNA的研究初报

 本文利用dsRNA特异性单克隆抗体,采用双抗体夹芯ELISA法,检测了栗疫菌中的dsRNA.结果表明,该法不仅能从栗疫菌低毒力菌株的总核酸粗提液中检测到dsRNA而不受其它核酸的干扰,而且比电泳法简便迅速,在大量筛选含dsRNA的栗疫菌低毒力菌株中具有很大的实用价值。     

Biological Control of Sclerotinia Diseases of Rapeseed by Aerial Applications of the Mycoparasite Coniothyrium minitans

Indoor and field experiments were conducted to evaluate the efficacy of applying the mycoparasite Coniothyrium minitans to the aerial parts of rapeseed plants at the flowering stage to control sclerotinia diseases caused by Sclerotinia sclerotiorum. Under controlled conditions, a petal inoculation technique was used to determine the effect of conidial suspensions of C. minitans on suppression of sclerotinia leaf blight. Results showed that C. minitans was effective in inhibiting infection initiated by ascospores of S. sclerotiorum on flower petals by restricting mycelial growth of the pathogen. Suppression of lesion development was related to the conidial concentration of C. minitans, with larger lesions at low concentration (5×10³conidia ml⁻¹), but smaller lesions at high concentration (5×10⁴ conidia ml⁻¹ or higher). When C. minitans-treated rapeseed leaves were inoculated with mycelia of S. sclerotiorum, C. minitans failed to prevent infection of leaves, but caused a significant reduction in number of sclerotia produced on the diseased leaves. No significant difference in efficacy was detected between the two isolates of C. minitans, LRC 2137 and Chy-1, on the two rapeseed cultivars, Westar (spring type) and Zhongyou 821 (winter type). Results of field trials showed a significant reduction of stem rot of rapeseed in four (1997, 1999, 2003 and 2004) out of five years by aerial application of C. minitans, compared with controls. No significant difference in suppressive efficacy was observed between the treatments of C. minitans (10⁶ conidia ml⁻¹), C. minitans (10⁶ conidia ml⁻¹) + benomyl (50 μg ml⁻¹) and benomyl (100 μg ml⁻¹) in 2003, and between the treatments of C. minitans (10⁶ conidia ml⁻¹), C. minitans (10⁶ conidia ml⁻¹) + vinclozolin (100 μg ml⁻¹) and vinclozolin (500 μg ml⁻¹) in 2004. Sclerotia of S. sclerotiorum collected from diseased plants in plots treated with C. minitans in 1999, 2000 and 2003, or with C. minitans + benomyl in 2003 were infected by C. minitans at frequencies ranging from 21.3 to 54.5%. This study concludes that aerial spraying of C. minitans is an effective method for controlling sclerotinia diseases of rapeseed.     

Capsid protein sequence gene analysis of <Emphasis Type="Italic">Apple mosaic virus</Emphasis> infecting pears

Apple mosaic virus (ApMV, genus Ilarvirus) was detected in pears, a previously non-reported virus host. No symptoms were visible on the hosts leaves. Seventeen out of 22 randomly selected pear trees in Italy (Lombardy) and in three regions in the Czech Republic were ApMV-infected. All nine newly sequenced ApMV isolates from pears had a 15-nucleotide insertion in the capsid protein gene in identical position of that of apple isolates compared with isolates from hop and prunes. The insertion is the most prominent (but not essential) modification of the capsid protein gene, which results in a phylogenetic separation of ApMV isolates into three clusters. Sequence analysis data of an additional 15 isolates revealed a sequence correlation with kernelled fruit trees (apple and pear).     

Soil solarization as a substitute for methyl bromide fumigation in greenhouse tomato production in Cyprus

Preplant soil fumigation with methyl bromide (MB) is presently standard practice in greenhouse tomato production. Since this compound is scheduled to be phased out by 2005, the possibility of using solarization as an alternative soil disinfestation method was examined in four greenhouse tomato trials. Solarization was applied for 8 weeks in July-August, using transparent polyethylene sheets for soil mulching, and compared with MB fumigation applied in September, before planting, at 80 g/m². Solarization raised the maximum soil temperature by 9°C and reduced the population density ofFusarium spp. in soil by 91–98%. Similar reductions of soil inoculum (95–99%) were obtained with MB fumigation. Both methods provided effective control of Fusarium wilt, Verticillium wilt and corky root rot on tomato plants. MB fumigation was in addition highly effective against root-knot nematodes, whereas nematode control with solarization did not exceed 50%. Both treatments resulted in similar fruit yield increases, ranging within 90–140% compared with plants grown in untreated soil. During the second cropping season following soil treatment, solarization exhibited two times higher residual effectiveness against vascular wilt diseases compared with MB fumigation. The latter treatment, however, was superior to solarization in its residual effectiveness against root-knot nematodes and to a lesser extent against corky root rot. Fruit yields from solarized and MB-fumigated soil during the second cropping season were higher than those obtained from untreated soil by approximately 35% and 60%, respectively. In Cyprus, solarization appears to be an effective alternative to MB fumigation in greenhouse tomato production, especially if integrated with other approaches enabling more effective nematode control.