Biology and control of white blister rust of Brassicaceae vegetables

This research project was a response to the outbreak of white blister rust disease and its spread to Brassica oleracea crops in Australia. Disease management strategies based on the understanding of the Albugo - Brassica pathosystem, epidemiology, and the use of chemical control measures were developed for the Australian Brassica industry.

Races of Albugo candida causing white blister rust on brassica vegetables in Australia

Albugo Candida races of Australian isolates from B. oleracea var. italica (broccoli), B. rapa var. pekinensis (pak choi) and var. chinensis (Chinese cabbage), and Capsella bursa-pastoris (shepherd's purse) were identified on a set of Brassicaceae hosts. Isolates from broccoli were identified as A. candida race 9 (Ac 9), from pak choi and Chinese cabbage as the sub-race V of Ac 7 or as a mixed population of sub-races A and V of the same race. Isolates from Shepherd's purse were identified as Ac 4. Australian Ac 9 isolates caused white blister disease on broccoli, broccolini, cauliflower, Brussels sprout and other related Brassicaceae hosts including B. nigra (black mustard), B. napus (oilseed rape), and on B. rapa (turnip rape) 'Torch' (a differential host of Ac 7). All cultivars of cabbage (B. oleracea var. capitata) and one new broccoli 'Booster' inoculated with isolates from broccoli were immune to the isolate tested. This result indicates that the Australian A. candida varies from the European one that causes disease on cabbage as well as on other B. oleracea varieties and additionally on shepherd's purse and an American one that causes disease on cabbage. The genotype of B. nigra tested was susceptible to both Ac 9 and Ac 7. This result indicates that B. nigra can serve as a host for both races. This study provides the first record of white blister disease on B. napus ('Hobson' and 'Regent') in Australia.

Molecular phylogenetic relationships between Albugo candida collections on the Brassicaceae in Australia

Recent phylogenetic analyses of Albugo candida using the mitochondrial cytochrome c oxidase subunit II (cox2) gene, the nuclear ribosomal RNA large subunit (LSU) gene and the nuclear ribosomal RNA internal transcribed spacer (ITS) gene region have revealed significant genetic variation and led to the description of new species in the A. candida complex. This study examined the genetic diversity within Australian collections of A. candida from various Brassicaceae species in a range of geographic locations. Phylogenetic analysis of 31 Australian A. candida collections from 11 hosts using the rDNA ITS region, rDNA LSU region and cox2 mtDNA showed that the majority of Australian A. candida collections were the common form of A. candida. One collection from a common weed host, hairy bitter cress (Cardamine hirsuta), was found to belong to a previously reported but undescribed species, while three collections, also from C. hirsuta, were found to belong to a new undescribed species.

Blister pack

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Studies on physiologic specialisation of albugo candida causing white blister on broccoli in Victoria

White blister caused by oomycete Albugo candida (Pers. Ex. Lev.) Kuntze, (AC), is an important disease affecting many cruciferous hosts, including vegetable brassicas. The outbreaks of white blister in broccoli and cauliflower crops (Brassica oleracea var. italica and var. botrytis) in Southern Australia in the last three years led to restrictions on movement of fresh produce and seedlings from the disease-affected areas. Current classification of AC races is based on physiologic specialisation of this pathogen. Race 9 has been identified to cause white blister on B. oleracea in the USA. We report on specialisation of AC causing disease in Victorian broccoli crops and the use of molecular tools for the separation of AC races. In a glasshouse, 12 Brassicaceae species/varieties replicated 6 times, were inoculated twice at the fully developed cotyledon stage with a distilled water suspension of zoosporangia (1x104 per ml) collected from a single broccoli leaf. Two weeks after inoculation the incidence of white blister on cotyledons and seedling leaves of cauliflower, broccoli, black mustard and Indian mustard was 79.7, 78.4, 73.7 and 6.9% respectively. Cabbage plants were symptomless indicating that further specialisation of the pathogen may have occurred in Australia. High disease incidence among black mustard plants shows that the Australian isolate differs from overseas AC race 9. The interaction of a number of B. oleracea varieties to a range of AC isolates from various hosts will be investigated. Degenerate primers are now being used to amplify actin and β-tubulin genes to identify race specific polymorphisms in AC isolates from three different hosts (wild radish, Chinese cabbage, and broccoli). Differing PCR amplification efficiencies from broccoli and wild radish isolates using degenerate actin primers indicates sequence differences in the two isolates. The fragments are now being cloned and sequenced for race comparison.

Multipurpose disinfecting solutions and their interactions with a silicone hydrogel lens

Purpose: To assess the compatibility of a new silicone hydrogel lens, asmofilcon A (with four multipurpose disinfecting solutions: OPTIFREE RepleniSH, ReNu MultiPlus, Solo-Care Aqua and MeniCare Soft). Ocular responses and subjective responses were monitored with each lens-care system combination.

Methods: The study was conducted as a prospective, bilateral, clinical trial with a single-masked investigator, and randomized cross-over design with four phases, (one for each care system). Each study phase comprised of two consecutive days of lens wear where the lenses were inserted on day 1 directly from the blister-packs and worn for over 8 hr, then inserted on day 2 after overnight disinfection with one of the study lens care systems. Twenty-five adapted soft contact lens wearers who were able to wear their habitual lenses comfortably for more than 12 hr were recruited.

Results: There were statistically significant differences in corneal staining found for all the lens-care systems when comparing the results of day 1 (from the blister pack) with day 2 (following care system use) (P < 0.05). ReNu MultiPlus solution had the highest grade for corneal staining at the 2-hr time point on day 2 which then decreased by 6 hr (P < 0.05). There was no difference between the lens care systems and the rating of subjective comfort over either of the two days. The rating of dryness and burning sensations were only slightly increased at 6 hr for all lens care systems except ReNu MultiPlus where burning was highest on insertion (P < 0.05).

Conclusion: Corneal staining observed in this study does not seem to have been related to the presence of polyhexamethylene biguanide (0.0001% wv) that was present in three of the four care systems. Only one care system (ReNu MultiPlus) demonstrated an associated level of corneal staining that was statistically significant; however, this was not considered to be of clinical relevance. These results suggest that using this novel surface-treated silicone hydrogel lens may result in less lens and lens care-related interactions.

Impact of a rub and rinse on solution-induced corneal staining

Purpose. To investigate whether the inclusion of a rub and rinse step before contact lens disinfection has an impact on solution-induced corneal staining.

Methods. This was a prospective, double-masked, single investigator study. Twenty participants were recruited for two visits, where balafilcon-A lenses were worn bilaterally for 2 h. Each pair of lenses was prepared using two different methodologies. The “control” lens was transferred from the blister pack directly into a storage case containing polyhexamethylene biguanide-based lens care solution. The contralateral “test” lens was rubbed and simultaneously rinsed using the same polyhexamethylene biguanide-based care solution, for either 60 s (visit 1) or 20 s (visit 2). Both lenses were then soaked in the solution overnight. After baseline corneal staining assessments, the lenses were inserted following a randomized contralateral model. After 2 h, lenses were removed, corneal staining was regraded, and comfort scores were obtained.

Results. Rubbed and rinsed test lenses induced significantly less corneal staining than control lenses for all participants during visit 1 (mean ± SD: 516 ± 843 vs. 2170 ± 902; p < 0.001) and visit 2 (522 ± 417 vs. 2091 ± 965; p < 0.001). There was no significant difference between the test lenses during visits 1 and 2 (p = 0.72) or controls (p = 0.50). Comfort scores did not differ between eyes (p > 0.05).

Conclusions. Corneal staining induced after 2 h of lens wear with the combination of balafilcon-A and polyhexamethylene biguanide-based lens care solution can be significantly reduced by including a rub and rinse step before overnight soaking. Further work is required to establish the longevity of this effect during the monthly wearing cycle.