A detached leaf bioassay to screen Durian cultivars for susceptibility to Phytophthora palmivora

A detached-leaf bioassay was developed and used to screen five durian (Durio zibethinus) cultivars against Phytophthora palmivora isolates from a trunk canker, root and fruit. The fruit isolate was less aggressive than the canker and root isolates. The bioassay using the canker isolate was later used to determine the variation in resistance of D. macarantha and nineteen cultivars of D. zibethinus. The cultivars displayed a range of responses with Parung and Gob being most tolerant, with Gaan Yaow, Chanee and Penang 88 being susceptible. The remaining germplasm fell between Gaan Yaow and Penang 88 in susceptibility. The leaf bioassay was found to be a rapid and reliable method for assessing the susceptibility of durian cultivars.

Comparative growth and pathogenicity of geographical isolates of Sclerotinia sclerotiorum on lentil genotypes

Isolates of Sclerotinia sclerotiorum were collected from infected lentil plants from 2 agro-ecological zones of Syria and used to study their comparative growth on culture media and pathogenicity on different lentil genotypes. The growth studies were carried out on Potato Dextrose Agar (PDA) growth media under laboratory conditions. Mycelial radial growth and sclerotial production were the parameters used to compare the isolates. Pathogenicity studies were carried out with selected isolates on 10 lentil genotypes, infected as detached shoots and as whole potted-plants in the plastic house. The isolates showed considerable variation in cultural characteristics through mycelial growth, mycelial pigmentation and sclerotial production in the media plates. There were significant differences in the growth and sclerotial production of most of the isolates, but no apparent correlation between mycelial growth and sclerotial production among the isolates. Genotype by isolate interactions was significant for the isolates tested for pathogenicity. These interactions, however, appeared to be caused by differences in virulence of the isolates and did not suggest the occurrence of distinct pathogenic races of the pathogen isolates.

Successful management of colletotrichum crown and stolon rot in runner production in sub-tropical Australia

Crown, stolon, and petiole rots caused by Colletotrichum gloeosporioides (C.g.) were first identified in runner beds of the Queensland Approved Runner Scheme (QARS) in February 1989. The outbreaks occurred annually from 1990 to 1994. Minor losses in subsequent fruit crops occurred from 1990 to 1993, with 50% post-establishment losses occurring on fruit farms in southeast Queensland in 1994. The objective of this work was to provide a control strategy for the disease that would give stability to the QARS. Runner-bed trials in 1993-1994 showed that Octave® (462 g/kg prochloraz as the MnCl2 complex) was highly effective in reducing the incidence field symptoms and laboratory recovery of C.g. from symptomless petioles. A simple detached petiole laboratory test for measuring fungicide efficacy in runner bed trials and for laboratory screening of fungicides, is described. Scheme protocols were changed to require that only foundation plants from tissue culture were allowed onto QARS sites. These were to be symptomless and to have tested negative for the presence of C.g. The application of Octave® at fortnightly intervals in all QARS nurseries has reduced the level of visible symptoms and the laboratory recovery of C.g. from symptomless petioles to almost zero.

Pest-damage relationships for Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) on vegetative soybean.

The response of vegetative soybean (Glycine max) to Helicoverpa armigera feeding was studied in irrigated field cages over three years in eastern Australia to determine the relationship between larval density and yield loss, and to develop economic injury levels. Rather than using artificial defoliation techniques, plants were infested with either eggs or larvae of H. armigera, and larvae allowed to feed until death or pupation. Larvae were counted and sized regularly and infestation intensity was calculated in Helicoverpa injury equivalent (HIE) units, where 1 HIE was the consumption of one larva from the start of the infestation period to pupation. In the two experiments where yield loss occurred, the upper threshold for zero yield loss was 7.51 ± 0.21 HIEs and 6.43 ± 1.08 HIEs respectively. In the third experiment, infestation intensity was lower and no loss of seed yield was detected up to 7.0 HIEs. The rate of yield loss/HIE beyond the zero yield loss threshold varied between Experiments 1 and 2 (-9.44 ± 0.80 g and -23.17 ± 3.18 g, respectively). H. armigera infestation also affected plant height and various yield components (including pod and seed numbers and seeds/pod) but did not affect seed size in any experiment. Leaf area loss of plants averaged 841 and 1025 cm2/larva in the two experiments compared to 214 and 302 cm2/larva for cohort larvae feeding on detached leaves at the same time, making clear that artificial defoliation techniques are unsuitable for determining H. armigera economic injury levels on vegetative soybean. Analysis of canopy leaf area and pod profiles indicated that leaf and pod loss occurred from the top of the plant downwards. However, there was an increase in pod numbers closer to the ground at higher pest densities as the plant attempted to compensate for damage. Defoliation at the damage threshold was 18.6 and 28.0% in Experiments 1 and 2, indicating that yield loss from H. armigera feeding occurred at much lower levels of defoliation than previously indicated by artificial defoliation studies. Based on these results, the economic injury level for H. armigera on vegetative soybean is approximately 7.3 HIEs/row-metre in 91 cm rows or 8.0 HIEs/m2.

Pathogenic variation of Alternaria species associated with leaf blotch and fruit spot of apple in Australia

Four Alternaria species groups (A. longipes, A. arborescens, A. alternata/A. tenuissima and A. tenuissima/A. mali) are associated with leaf blotch and fruit spot of apple in Australia. There is no information on the variability of pathogenicity among the species and isolates within each species causing leaf blotch or fruit spot. We used a detached leaf assay and an in planta fruit inoculation assay to determine the pathogenicity and virulence of the four Alternaria species. Our results showed that isolates within the same species were not specific to either leaf or fruit tissue and showed great variability in pathogenicity and virulence, indicating cross-pathogenicity, which may be isolate dependent rather than species dependent. Generally, virulence of A. tenuissima and A. alternata isolates on leaf and fruit was higher than other species. Isolates of all species groups were pathogenic on leaves of different cultivars, but pathogenicity on fruit of different cultivars varied among isolates and species. Implications of our findings on prevalence of the diseases in different apple-producing regions in Australia and the development of targeted disease management of the diseases are discussed

Infection of mungbean seed by Macrophomina phaseolina is more likely to result from localized pod infection than from systemic plant infection

The ubiquitous fungal pathogen Macrophomina phaseolina is best known as causing charcoal rot and premature death when host plants are subject to post-flowering stress. Overseas reports of M.phaseolina causing a rapid rot during the sprouting of Australian mungbean seed resulted in an investigation of the possible modes of infection of seed. Isolations from serial portions of 10 mungbean plants naturally infected with the pathogen revealed that on most plants there were discrete portions of infected tissue separated by apparently healthy tissue. The results from these studies, together with molecular analysis of isolates collected from infected tissue on two of the plants, suggested that aerial infection of aboveground parts by different isolates is common. Inoculations of roots and aboveground parts of mungbean plants at nine temperaturexsoil moisture incubation combinations and of detached green pods strongly supported the concept that seed infection results from infection of pods by microsclerotia, rather than from hyphae growing systemically through the plant after root or stem infection. This proposal is reinforced by anecdotal evidence that high levels of seed infection are common when rainfall occurs during pod fill, and by the isolation of M.phaseolina from soil peds collected on pods of mungbean plants in the field. However, other experiments showed that when inoculum was placed within 130mm of a green developing pod and a herbicide containing paraquat and diquat was sprayed on the inoculated plants, M.phaseolina was capable of some systemic growth from vegetative tissue into the pods and seeds.

Commercial-scale Alternaria brown spot resistance screening as the first step in breeding new mandarins for Australia

Rapid screening tests and an appreciation of the simple genetic control of Alternaria brown spot (ABS) susceptibility have existed for many years, and yet the application of this knowledge to commercial-scale breeding programs has been limited. Detached leaf assays were first demonstrated more than 40 years ago and reliable data suggesting a single gene determining susceptibility has been emerging for at least 20 years. However it is only recently that the requirement for genetic resistance in new hybrids has become a priority, following increased disease prevalence in Australian mandarin production areas previously considered too dry for the pathogen. Almost all of the high-fruit-quality parents developed so far by the Queensland-based breeding program are susceptible to ABS necessitating the screening of their progeny to avoid commercialisation of susceptible hybrids. This is done effectively and efficiently by spraying 3-6 month old hybrid seedlings with a spore suspension derived from a toxin-producing field isolate of Alternaria alternate, then incubating these seedlings in a cool room at 25°C and high humidity for 5 days. Susceptible seedlings show clear disease symptoms and are discarded. Analysis of observed and expected segregation ratios loosely support the hypothesis for a single dominant gene for susceptibility, but do not rule out the possibility of alternative genetic models. After implementing the routine screening for ABS resistance for three seasons we now have more than 20,000 hybrids growing in field progeny blocks that have been screened for resistance to the ABS disease.

Genome-wide association analysis and differential expression analysis of resistance to Sclerotinia stem rot in Brassica napus

Brassica napus is one of the most important oil crops in the world, and stem rot caused by the fungus Sclerotinia sclerotiorum results in major losses in yield and quality. To elucidate resistance genes and pathogenesis-related genes, genome-wide association analysis of 347 accessions was performed using the Illumina 60K Brassica SNP (single nucleotide polymorphism) array. In addition, the detached stem inoculation assay was used to select five highly resistant (R) and susceptible (S) B. napus lines, 48 h postinoculation with S. sclerotiorum for transcriptome sequencing. We identified 17 significant associations for stem resistance on chromosomes A8 and C6, five of which were on A8 and 12 on C6. The SNPs identified on A8 were located in a 409-kb haplotype block, and those on C6 were consistent with previous QTL mapping efforts. Transcriptome analysis suggested that S. sclerotiorum infection activates the immune system, sulphur metabolism, especially glutathione (GSH) and glucosinolates in both R and S genotypes. Genes found to be specific to the R genotype related to the jasmonic acid pathway, lignin biosynthesis, defence response, signal transduction and encoding transcription factors. Twenty-four genes were identified in both the SNP-trait association and transcriptome sequencing analyses, including a tau class glutathione S-transferase (GSTU) gene cluster. This study provides useful insight into the molecular mechanisms underlying the plant's response to S. sclerotiorum.