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.     

Identification of Alternaria spp. on wheat by pathogenicity assays and sequencing

The pathogenicity of Alternaria spp. isolated from wheat leaves collected in regions where alternaria leaf blight has been reported was compared with that of IMI reference isolates of A. triticina and A. alternata using two durum and two bread wheat genotypes. To identify isolates putatively corresponding to A. triticina , morphological and DNA sequence analyses based on ribosomal DNA from the internal transcribed spacer (ITS) region (ITS1, 5·8S rRNA gene, ITS2) and toxicity bioassays of culture filtrate were combined. Glasshouse inoculations provided reliable information to assess the pathogenicity of A. triticina isolates on wheat. Alternaria leaf blight symptoms were produced by the A. triticina isolates only on durum wheat cv. Bansi, while A. alternata , A. tenuissima and A. arborescens isolates were found to be nonpathogenic on the wheat cultivars tested. Alternaria triticina isolates were distinguished from other Alternaria species by Simmons and Roberts' sporulation pattern 6 and two to three conidia per sporulation unit associated with primary conidia bearing long (> 7  µ m) apical secondary conidiophores. Phylogenetic analysis also proved effective at discriminating wheat-pathogenic A. triticina from other nonpathogenic Alternaria species. Alternaria triticina isolates yielded longer ITS sequences than A. alternata , A. tenuissima and A. arborescens isolates, leading to clear-cut differences as visualized with agarose gel electrophoresis. Additionally, only culture filtrates of A. triticina isolates caused nonspecific necrotic lesions on leaves of 3-week-old wheat plants.     

A Severe Hellenic CMV Tomato Isolate: Symptom Variability in Tobacco, Characterization and Discrimination of Variants

A severe strain of Cucumber mosaic virus (CMV) originating from an infected tomato plant (Gastouni-Olympia, Greece) was isolated in tobacco (Nicotiana tabacum cv. Xanthi nc), after three serial local lesion passages in Chenopodium quinoa and designated CMV-G. CMV-G induces yellow mosaic (YM) symptoms in tobacco. When CMV-G was passed mechanically through C. quinoa, phenotypic variants inducing YM or green mild mosaic (MM) in tobacco were isolated. Aphid transmission, from different hosts, appears to be an effective approach for separating MM variants of CMV-G from YM variants. In particular, aphid transmission from zucchini proved to be very efficient in selecting for MM variants. In contrast, aphids transmitted only YM variants from tomato plants. Molecular characterization of CMV-G and its progeny resulted in their classification in the CMV subgroup IB, free of satellite RNA, being the first discovery of the subgroup IB in Greece. In the Solanaceae family (tobacco, tomato, pepper) YM variants induced more severe symptoms than the MM variants. YM and MM phenotype was stable in tobacco for all seven passages tried using the obtained YM and MM variants. Cross-protection experiments showed that an isolated MM variant was able to protect tobacco plants against a challenge infection by a YM variant.     

Development of specific PCR primers for identification and detection of <Emphasis Type="Italic">Phytophthora capsici</Emphasis> Leon

A PCR-based method was developed for the identification and detection of Phytophthora capsici in pepper plants. Three PCR primers (CAPFW, CAPRV1 and CAPRV2) specific for P. capsiciwere designed based on the sequence of its internal transcribed spacer regions. CAPFW/CAPRV1 amplify a 452 bp product from P. capsici DNA whereas CAPFW/CAPRV2 a 595 bp fragment; neither set amplifies DNA from pepper or several fungi pathogenic to pepper. In conventional (single-round) PCR, the limit of detection was 5 pg DNA for both primer sets, whereas in nested PCR the detection limit for both was of 0.5 fg. However, when the dilution series of target DNA were spiked with plant DNA, amplification declined two-fold in both conventional and nested PCR. The CAPFW/CAPRV2 set in conventional PCR was used to detect P. capsici DNA in inoculated plants. Detection occurred as soon as 8h post-inoculation in stem samples from infected but still symptomless plants. The method was also tested to detect fungal DNA in infected soils.     

Fusarium head blight and mycotoxin contamination of wheat,a review

Summary An infection of bread wheat by fusarium head blight contaminates the crop with mycotoxins, particularly deoxynivalenol (DON) and nivalenol (NIV). The toxicity and natural occurrence of these mycotoxins in wheat are reviewed. Based on 8 years data of fusarium head blight epidemics of wheat in the Netherlands, DON contamination of the grain was estimated. Fusarium head blight ratings averaged an infection of 1.7% of all spikelets; estimates for DON contamination averaged 0.9 mg kg^–1. Taking a guideline level for DON in uncleaned bread wheat of 2 mg kg^–1, in 1979 and 1982 a wheat crop was produced with estimated DON concentrations above the limit of tolerance. Human and animal exposure to mycotoxins in the Netherlands appears to be small but chronic. The information presented in this paper illustrates the need for an annual evaluation of the crop for fusarium head blight incidence and mycotoxin content, and the necessity of fusarium head blight resistant wheat cultivars.Samenvatting Aaraantasting van tarwe doorFusarium culmorum enFusarium graminearum leidt tot vorming van mycotoxinen in het graan, waarvan deoxynivalenol (DON) en nivalenol (NIV) de belangrijkste toxinen zijn. In dit artikel wordt een overzicht gegeven van de toxicologische aspecten, en het voorkomen van deze toxinen in tarwe. Informatie over DON en NIV in tarwe in West-Europa is schaars. Gebaseerd op gegevens vanFusarium epidemieën in de jaren 1979–1986 wordt een schatting gegeven van de concentratie DON in Nederlandse tarwe. Rekening houdend met de herkomst en verwerking van tarwe, blijken zowel in dierlijk als menselijk voedsel lage concentraties DON chronisch voor te komen. Op basis van een maximaal toelaatbare dagelijkse dosis DON van 3 g kg^–1 lichaamsgewicht is de schatting van de dagelijkse opname van DON in het jaar volgend op de oogst van 1982 net op de grens. Zowel een jaarlijkse inventarisatie vanFusarium aantasting en DON besmetting van het graan, als de ontwikkeling vanFusarium-resistente rassen zijn noodzakelijk.     

Genetic diversity of Pueraria lobata (kudzu) and closely related taxa as revealed by inter-simple sequence repeat analysis

Kudzu (Pueraria lobata) is a noxious weed infesting some areas of the USA. Knowledge of its genetic variation in both native and invasive areas can lead to effective biological control measures. Inter-simple sequence repeat (ISSR) variations were studied in P. lobata and its four closely related congeneric species (P. edulis, P. montana, P. phaseoloides and P. thomsoni). ISSR results allowed a clear separation of these five species. For P. lobata, 108 plants from China and USA were analysed. The samples from the US were genetically closer to Chinese P. lobata populations than to other congeneric populations. High genetic differentiation was found for P. lobata, P. montana and P. thomsoni in Chinese samples. High genetic diversity and low population differentiation was found in P. lobata samples of the US. This supports the hypothesis of multiple introductions into the USA from different sources in Japan or China, followed by subsequent gene exchange and recombination.     

The effect of diflubenzuron on Eggs of the vine Moth,Lobesia (Polychrosis) Botrana Den. & Schiff (Lepidoptera: Tortricidae)

The activity of diflubenzuron against eggs ofLobesia botrana Den. & Schiff. was assayed with a dipping procedure, at 22° and 27° C. The compound was much more active at 27°C (LC-50 = 0.0072%) than at 22°C (LC-50 = 0.195%).     

Response of black, cranberry, kidney, and white bean to linuron

Dry bean producers in Ontario, Canada, have few herbicide options available for annual broad-leaved weed management and there is little information on the tolerance of dry bean to linuron. The tolerance of black, cranberry, kidney, and white bean to the pre-emergence (PRE) application of linuron at the rates of 0, 500, 1000, 1500, 2000, and 2500 g ai ha⁻¹ was evaluated in field studies conducted in 2005 and 2006 at Exeter and in 2006 at Ridgetown, Ontario. The four market classes differed in their response to linuron. Cranberry and kidney bean were more tolerant to the PRE application of linuron than black and white bean. Linuron applied PRE caused as much as 12% injury in cranberry and kidney bean, 47% injury in black bean, and 56% injury in white bean. Linuron applied PRE at 500–2500 g ai ha⁻¹ had no effect on the height of cranberry and kidney bean but decreased the height by 7, 8, and 15% in black bean and by 10, 13, and 23% in white bean at 1500, 2000, and 2500 g ai ha⁻¹, respectively. Linuron applied PRE at the rates evaluated did not cause any adverse affect on the yield of cranberry, kidney, and white bean but black bean yield was reduced by 16% at 2500 g ai ha⁻¹. Based on these results, there is not an adequate margin of crop safety for the PRE application of linuron in black and white bean at rates >1000 g ai ha⁻¹. However, there is a potential for the use of linuron PRE for weed management in cranberry and kidney bean at the rates evaluated.     

Removal of flowers or inflorescences affects ‘Barnea’ olive fruitlet post-anthesis abscission

A typical olive (Olea europaea L.) inflorescence consists of about 20 flowers. However, in many cultivars, only one fruit develops. This is due to massive abscission of flowers and fruitlets, which occurs during the first month after anthesis. In this study, we used the olive cultivar 'Barnea' to characterize the abscission mechanism and to try to increase fruit set by increasing the number of developed fruit per inflorescence. Removing the lateral flowers 3 weeks before anthesis increased fruit set by more than 50%. Removing all inflorescences but one from a branch increased the number of developed fruits from 0.93 to 2.8 during 2017 and from 0.91 to 3.34 fruits per inflorescence in 2018. Sugar quantification in the pistil revealed that starch level is high on the day of anthesis and low 25 days later in abscised as well as in developed fruit. Soluble carbohydrates are low on the day of anthesis, low in abscised flowers/fruitlets 25 days after anthesis and high in developed fruit. Screening the natural variation found in the Israeli germplasm collection revealed that in most cultivars less than one fruit per inflorescence has developed. However, there are unique cultivars with a higher fruit set.