Observations of Phytophthora spp. in Water Recirculation Systems in Commercial Hardy Ornamental Nursery Stock

Samples of water and sediment were taken from drains, reservoirs and wells from four commercial hardy ornamental nurseries with water recirculation systems. The samples were taken on seven different dates throughout a single year from August 1994 to July 1995. The samples were screened for Phytophthora species using five different methods: direct plating, three bait tests (using lupin seedlings, apples and Rhododendron leaves) and a DAS-ELISA (double-antibody sandwich enzyme-linked immunosorbent-assay) with two antisera. In the nurseries with old water recirculation systems, Phytophthora species were detected in the drains and in the reservoirs. In the nursery with a new recirculation system, the pathogens were only present in the drains. None of the water samples from wells in any of the nurseries were contaminated. Phytophthora species were present in the water as well as in the sediment samples from drains and reservoirs. They were detected in the water recirculation systems irrespective of the season. The number of isolates increased about sevenfold between late summer and spring. At least 12 different Phytophthora species were identified: some isolates were previously unrecorded species. The epidemiology of the pathogens in outdoor water recirculation systems as well as the importance of the results for commercial nurseries is discussed.     

Aromatic Substances Inhibiting Wheat Powdery Mildew Produced by a Fungus Detected with a New Screening Method for Phylloplane Fungi

Microorganisms isolated from wheat leaf surfaces were screened for inhibition of wheat powdery mildew. A new screening method, in which wheat leaves were inoculated with Blumeria graminis f. sp. tritici and incubated with the cultured microorganisms under non-contact conditions, was developed in the present study. Using this method, 10 phylloplane fungi that inhibited wheat powdery mildew were selected from 408 microorganisms isolated from wheat leaf surfaces. Among these 10 strains, a fungus designated as Kyu-W63 had an especially strong inhibitory effect. Kyu-W63 produced white colonies without spores when cultivated on PDA. Kyu-W63 had a strong aromatic odor when being cultured. Wheat powdery mildew was suppressed even though a membrane filter with a pore size of 0.45 μm was placed between the mycelial colony and wheat leaf segment. However, when activated charcoal was introduced, Kyu-W63 did not inhibit growth of B. graminis. It was presumed that volatile substances were involved in the inhibitory effect of Kyu-W63. GC-MS analysis was used to identify two substances produced by Kyu-W63 with molecular weights of 164 and 166. Kyu-W63 also inhibited the in vitro growth of four plant pathogenic fungi other than B. graminis. Received 19 September 2001/ Accepted in revised form 7 February 2002     

Ultrastructural Localization of Hydrogen Peroxide in Host Leaves Treated with AK-toxin I Produced by Alternaria alternata Japanese Pear Pathotype

The rapid generation of reactive oxygen species (ROS), called the oxidative burst, is one of the earliest host responses to pathogen infection or elicitor treatments. Therefore, we looked for the induction of ROS generation in Japanese pear leaves by the host-specific toxin, AK-toxin I using a cytochemical method for detecting H₂O₂. A small amount of non-specific generation of H₂O₂ was found in the cell walls in toxin- and water-treated susceptible and resistant leaves. Thus, the generation of H₂O₂ at cell walls appears to be caused by wounding stress during sampling. Specific generation of ROS, however, was found only in the membrane fragments and extended desmotubules characteristic of modified sites of the plasma membrane in the toxin-treated susceptible leaves. In addition, generation of H₂O₂ at plasma membranes was observed with higher frequency in toxin-treated susceptible leaves. This result indicates that the H₂O₂ generation was associated with damaged sites in the plasmalemma after toxin treatment and perhaps with the formation of membrane fragments from altered portions of the invaginated plasma membrane. Received 21 September 2001/ Accepted in revised form 25 October 2001     

The Complete Nucleotide Sequence of a Japanese Isolate of White clover mosaic virus Strain RC

The complete nucleotide sequence was determined for genomic RNA of White clover mosaic virus (WClMV-RC) isolated from red clover (Trifolium pratense) in Japan, It is 5843 nucleotides in length, excluding the poly(A) tail at the 3' terminus. Similar to other potexviruses, it contains five open reading frames (ORFs 1 through 5), which putatively encode an RNA-dependent RNA polymerase (RdRp) (147 kDa), a triple gene block (TGB) (26 kDa/13 kDa/7 kDa), and a coat protein (CP) (22 kDa), respectively. The deduced amino acid sequence of the WClMV-RC CP was identical to that of WClMV-O, one of two New Zealand isolates, but only 85% identical to that of WClMV-M, the other New Zealand isolate, because of heterogeneity in the C-termini of CP amino acid sequences. The implication of this CP heterogeneity is discussed. Received 30 August 2001/ Accepted in revised form 11 January 2002     

Variation in the development of secondary dormancy in oilseed rape genotypes under conditions of stress

Summary A substantial amount of seed is left in the fields before and during harvest of oilseed rape. Although this crop exhibits little or no primary dormancy, the absence of certain environmental cues that promote germination of imbibed seeds induces secondary dormancy. The work reported investigated the extent to which environmental stress conditions, including osmotic stress, low oxygen stress and anaerobiosis, induce secondary dormancy in oilseed rape, and examined the variation in development of secondary dormancy between and within genotypes. Osmotic stress was most effective in inducing dormancy. Anaerobic treatment produced very few dormant seeds, as did an atmosphere low in oxygen and high in nitrogen. The development of secondary dormancy under osmotic stress varied considerably between and within genotypes. Dormancy ranged from almost zero to about 60% for winter genotypes and about 85% for spring types. Within genotypes, variations occurred between seed lots and years of harvest. Temperature variations affected the percentage of dormant seeds. More dormant seeds were likely to be produced with incubation under water stress at 20 °C than at 12 °C. In winter genotypes, fewer dormant seeds were produced when incubation temperature and germination test temperatures differed. Thus, incubating at 20 °C and 12 °C, followed by germination tests at 20 °C and 12 °C, respectively, produced most dormant seeds. Also, in the winter genotypes, the potential development of secondary dormancy was positively correlated with the pattern and speed of germination of untreated seeds.     

Movement of Xanthomonas campestris pv. vitians in the stems of lettuce and seed contamination

Xanthomonas campestris pv. vitians , the causal agent of bacterial leaf spot of lettuce (BLS), can be seedborne, but the mechanism by which the bacteria contaminates and/or infects lettuce seed is not known. In this study, the capacity of X. campestris pv. vitians to enter and translocate within the vascular system of lettuce plants was examined. The stems of 8- to 11-week-old lettuce plants were stab-inoculated, and movement of X. campestris pv. vitians was monitored at various intervals. At 4, 8, 12 and 16 h post-inoculation (hpi), X. campestris pv. vitians was recovered from 2 to 10 cm above (depending on stem length) and 2 cm below the inoculation site. Xanthomonas campestris pv. vitians was also recovered from surface-disinfested stem sections of spray-inoculated plants. Together, these results are consistent with X. campestris pv. vitians invading and moving systemically within the vascular system of lettuce plants. To investigate the mechanism of seed contamination, lettuce plants at the vegetative stage of growth were spray-inoculated with X. campestris pv. vitians and allowed to develop BLS. Seed collected from these plants had a 2% incidence of X. campestris pv. vitians external colonization, but no bacteria were recovered from within the seed.     

Genetic variability within Phaeoisariopsis griseola from Central America and its implications for resistance breeding of common bean

The genetic and virulence variability of 112 isolates of Phaeoisariopsis griseola , collected from various locations in Central America, were studied using seven random amplified polymorphic DNA (RAPD) primers and 12 common-bean differential genotypes. Broad molecular diversity ( H  = 0·92) among isolates was found using RAPD markers. Fifty pathotypes were identified on 12 differential bean genotypes, 29 of which were represented by only one isolate. Only 18 pathotypes were found in two or more countries. Pathotype 63-63 was the most virulent and caused leaf spots on all 12 common-bean differential genotypes. Comparison of virulence phenotypes and RAPD profiles to known Andean P. griseola isolates confirmed that all isolates belonged to the Mesoamerican group. Pairwise comparison between individual RAPD loci showed that the majority were in gametic phase linkage disequilibrium, revealing that P. griseola maintains a genetic structure that is consistent with asexual reproduction. The molecular and virulence diversities of P. griseola isolates from Central America imply that using single resistance genes to manage angular leaf spot is inadequate and stacking resistance genes may be necessary to manage the disease effectively.     

Effects of aqueous extracts and decomposition of Mikania micrantha H.B.K. debris on selected agronomic crops

Laboratory and greenhouse experiments were conducted to determine the effects of aqueous extract and the debris of Mile-a-Minute ( Mikania micrantha H.B.K.) on the germination and growth of four test species, viz. tomato ( Lycopersicon esculentum Mill.), Chinese cabbage ( Brassica chinensis L.), corn ( Zea mays L.) and long bean ( Vigna sesquipedalis L.). Germination percentage, radicle length and fresh weight of two crops, Chinese cabbage and tomato, decreased progressively when plants were exposed to increasing concentrations (12.5, 25, 37.5 and 50 g L⁻¹) of aqueous extract of Mikania , but did not affect those of corn and long bean seedlings. The emergence of tomato and Chinese cabbage was greatly reduced when Mikania debris was incorporated into the soil, irrespective of the amount of debris and the decomposition period. In contrast, the emergence of corn and long bean seedlings was not affected. The application of fertilizer (NPK 15 : 15 : 15) enhanced the fresh weight of all seedlings tested as compared with those not supplied with the fertilizer. Four compounds were identified in the leaf extract of Mikania , viz. caffeic acid, p -hydroxybenzaldehyde, resorcinol and vanillic acid.