Use of real-time PCR to examine the relationship between disease severity in pea and <Emphasis Type="Italic">Aphanomyces euteiches</Emphasis> DNA content in roots

Aphanomyces euteiches causes severe root rot of peas. Resistance is limited in commercial pea cultivars. Real-time fluorescent PCR assay specific for A. euteiches was used to study the relationship between disease severity and pathogen DNA content in infected peas. Five pea genotypes ranging in levels of resistance were inoculated with five isolates of A. euteiches. Plants were visually rated for disease development and the amount of pathogen DNA in roots was determined using the PCR assay. The susceptible genotypes Genie, DSP and Bolero tended to have significantly more disease and more pathogen DNA than the resistant genotypes 90-2079 and PI 180693. PI 180693 consistently had less disease, while 90-2079 had the lowest amount of pathogen DNA. The Spearman correlation between pathogen DNA quantity and disease development was positive and significant (P < 0.05) for three isolates, but was not significant for two other isolates. This suggests that the real-time PCR assay may have limited application as a selection tool for resistance in pea to A. euteiches. Its utility as a selection tool would be dependent on the correlation between disease development and pathogen DNA content for a given pathogen isolate. The accuracy and specificity of the real-time PCR assay suggests considerable application for the assay in the study of mechanisms of disease resistance and the study of microbial population dynamics in plants.     

Effects of shading and NO3:NH4 ratio on the yield,quality and N metabolism in strawberry

The effect of 50% shading and NO₃:NH₄ ratio (0:100, 75:25, 50:50, and 25:75) in the nutrient solution on growth, yield, quality and N metabolism in hydroponically grown strawberry (Fragaria × ananassa var Camarosa) was evaluated. Both fresh and dry weights of leaves were significantly lower when a high concentration of either NO₃ (100%) or NH₄ (75%) was the sole N source in the nutrient solution. In unshaded plants, increasing of both NH₄ and NO₃ ratio in the nutrient solution reduced photosynthetic (Pn) rate, however in shaded plants the reduction of Pn became more pronounced at a higher ratio of NH₄ in the nutrient solution. The yield in terms of fresh and dry weight of fruit per plant was significantly increased at the 75:25 and 50:50 (NO₃:NH₄) treatments. Fruit size was significantly affected by the treatments, so that the biggest fruits in both shaded and unshaded plants were obtained under the 75:25 and 50:50 (NO₃:NH₄) treatments. Total soluble solid (TSS) in unshaded plants was increased with increasing NH₄ ratio in the nutrient solution, however in shaded plants it was reduced at high NH₄ ratio in the nutrient solution. In both shaded and unshaded plants, higher concentration of NH₄ significantly reduced the post-harvest life of the fruits. The increase of tissue N concentration was nearly proportional to the NH₄ concentration in the nutrient solution. The activity of nitrate reductase (NR) was increased by increasing NH₄ from 0 to 50% and then reduced at a higher ratio of NH₄ in the solution. Shading increased NH₄ concentration so that the shaded plant had nearly twice as high NH₄ concentration in the leaves. The increase of NH₄ concentration induced by shading could be partially the reduction of NH₄ assimilate because of the shortage of carbohydrate.     

Intra-specific variation and inheritance of BYDV-PAV transmission in the aphid <Emphasis Type="Italic">Sitobion avenae</Emphasis>

Vector efficiency of 44 clonal lines (clones) of Sitobion avenae belonging to 31 different genotypes (distinct patterns for five microsatellite loci) originating from Western France was evaluated by transmitting the isolate PAV4 of BYDV-PAV to barley seedlings. Variation in transmission rates from 3.7% to 92.5% was observed, with significant effects of the aphid clone, of the plant species on which clones were collected, and of the reproductive mode of the clones. When genotypes are considered instead of clones, a continuum in transmission rates was observed. A subset of S. avenae clones was tested for transmission of one (10 clones) and 13 (4 clones) other BYDV-PAV isolates, and a clear clone effect modulated by an isolate effect was observed. Crosses were made between clones with different vectoring phenotypes and their F1 progeny were tested for PAV4 transmission. The narrow sense heritability of the PAV transmission character was rather high in the F1 families (h²=0.5) and the segregation analyses suggested an oligo/polygenic determinism of this character. The possibility of generating new transmission variants by sexual reproduction and its consequences on transmission mechanism studies and on BYD epidemics are discussed.     

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.     

Implications of on-farm research for local knowledge related to fruit flies and the weaver ant Oecophylla longinoda in mango production

We interviewed half of the mango-growers in northern Benin, including 15 farmers involved in a regional fruit fly project, and held focus group discussions with women fruit-pickers. They were asked about pest management and their knowledge of a weaver ant, Oecophylla longinoda. All considered low yields due to fruit flies to be the principal constraint upon mango production, estimating economic losses to be between 20 and 45%. None could recognize damage during the first 2 days after fruit fly egg deposition. On-farm research persuaded farmers to stop using insecticides and it also changed negative perceptions of Oecophylla. Over 80% of the farmers involved in on-farm research, compared to 25% of those not involved, reported Oecophylla to be beneficial. All fruit-pickers knew that ants protected mango from fruit flies, with 60% attributing better mango quality in terms of appearance, shelf-life and sweetness to the presence of Oecophylla. Nevertheless, 40% of the pickers still considered weaver ants a nuisance pest during harvest. Ways of reducing this nuisance need to be developed for Oecophylla to gain wider acceptance by mango-growers.     

Storage performance of clementines treated with hot water,sodium carbonate and sodium bicarbonate dips

Summary

The potential of using hot water (2.5 min at 45°C), 2% sodium carbonate or 2% sodium bicarbonate solutions alone or combined with hot water, for control of Penicillium digitatum (green mould) was investigated on commecially ripe clementines. The effect of such treatments on the post-harvest storage life of clementines was determined during two months analysing weight loss, increase in rind deformation and internal maturity. Carbonate and bicarbonate solutions effectively control green mould during storage but hot water does not. Compared with the control and hot-water treated fruits, clementines treated with carbonate and, to a lesser extend with bicarbonate, showed higher weight loss and lower firmness during storage. In contrast, lower increase in ripening index was found when the fruits were treated with carbonate or bicarbonate. Fruits treated with hot water alone or combined with carbonate exhibited no change in ripening during the first month of storage. Rinsing the fruit significantly reduces weight and firmness loss in the fruits treated with carbonate and bicarbonate, but did not change the ripening index. The primary finding of this work was that carbonate and bicarbonate treatments might significantly increase the loss in weight and firmness during storage. Rinsing the fruit immediately after treatment appeared to be a simple solution to avoid these losses in quality.     

Variable response of open-pollinated seedling progeny of avocado to Phytophthora root rot

Phytophthora root rot, caused byPhytophthora cinnamomi Rands, is the most important disease of avocado (Persea americana Miller). In an attempt to identify root rot-resistant rootstocks that could ultimately be used under conditions in southern Florida, we screened open-pollinated progeny of avocado from the National Germplasm Repository in Miami. From 1996 to 1998, a total of 2,355 seedlings from 51 accessions were examined in potting mix artificially infested withP. cinnamomi. Most seedlings developed severe root rot, but tolerance was observed in some families (i.e., progeny of certain accessions). Although the most susceptible families developed mean disease ratings of up to 97% root necrosis, mean ratings for the most tolerant families were less than 60%. There was also a strong relationship between the racial background of the female parent and the tolerance of seedlings. Seedlings of the West Indian race and hybrids between it and the Guatemalan race were significantly more tolerant than those from other parents (P< 0.05). Individuals in several families developed < 50% root necrosis, the arbitrary standard of tolerance in this study. Twelve families accounted for 82% (188 of 229) of the tolerant seedlings, and only two of these did not have a West Indian or Guatemalan × West Indian pedigree. Broad-sense heritability for PRR tolerance was 0.45. This is the first report on the inheritance of PRR tolerance in avocado and on the influence of genotype and racial pedigree under controlled conditions.     

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.     

Evaluation of non-chemical seed treatment methods for the control of Alternaria dauci and A. radicina on carrot seeds

The current study was initiated to evaluate the efficacy of physical methods (hot water, aerated steam, electron treatment) and agents of natural origin (resistance inducers, plant derived products, micro-organisms) as seed treatments of carrots for control of Alternaria dauci and A. radicina. Control of both Alternaria species by seed treatment with the resistance inducers was generally poor. Results were also not satisfactory with most of the formulated commercial micro-organism preparations. Based on the average of five field trials, one of these, BA 2552 (Pseudomonas chlororaphis), provided a low but significant increase in plant stand. Among the experimental micro-organisms, the best results were obtained with Pseudomonas sp. strain MF 416 and Clonostachys rosea strain IK726. A similar level of efficacy was provided by seed treatment with an emulsion (1%) of thyme oil in water. Good and consistent control was generally achieved with the physical methods aerated steam, hot water and electron treatment. Aerated steam treatment was, apart from the thiram-containing chemical standard, the best single treatment, and its performance may at least partially be due to extensive pre-testing, resulting in dosages optimally adapted to the respective seed lot. In some of the experiments the effect of the hot water treatment, which was tested at a fixed, not specifically adapted dosage, was significantly improved when combined with a Pseudomonas sp. MF 416 or C. rosea IK726 treatment. The results are discussed in relation to the outcome of experiments in which the same seed treatment methods and agents were tested in other seed-borne vegetable pathosystems.