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Management of postharvest diseases of sub-tropical and tropical fruit using their natural resistance mechanisms
Project Leader
Dr Lindy Coates

Email
lindy.coates@deedi.qld.gov.au

Fax
0738462387

Phone
0732554353

Project Country
Philippines [1]

Inactive project countries

Sri Lanka

Project Coordinator Phone

(02) 6217 0553

Project Outcomes
The project team tested activators that were known resistance-inducing agents, including acibenzolar-S-methyl (Bion), and elicitors derived from fungal pathogens (in banana). In another component they characterised some key biochemical defences contributing to the resistance, and identified treatments, varietal properties or other agronomic practices which may influence

their relative concentrations. Mango was the common crop studied in Australia, Sri Lanka and the Philippines. In field trials, Bion was the consistently effective activator of resistance to anthracnose disease, when applied as a foliar spray or as a soil drench 3-5 times throughout the fruiting period. There is clearly the potential for reducing the number of fungicides applied in a given season if Bion is applied. Ultraviolet (UV-C) treatment to harvested mango activated biochemical defences and reduced anthracnose. Another key finding from field trials in Australia (cv. Keitt) and Sri Lanka (cv. Karuthacolomban) was that increasing nitrogen fertilisation enhances anthracnose levels in fruit, which was correlated with high skin nitrogen and lower levels of preformed alk(en)ylresorcinols in skin tissue. (The capacity to analyse the alk(en)ylresorcinol compounds was made possible after establishing collaboration with an expert in Poland.) Some mango cultivars (and rootstocks) consistently showed high levels of resistance to anthracnose, e.g. 'Keitt' in Australia and 'Gira' and 'Karuthacolomban' in Sri Lanka. There was some correlation between resistance amongst varieties and levels of constitutive defences. In Sri Lanka the galloyltannin class of compounds was identified as a major component contributing to antifungal activity in mango peel extracts. All banana work was conducted in Sri Lanka. The existence and partial characterisation of several phenylphenalone-type phytoalexins accumulating in response to infection with Phyllosticta musarum, the pathogen causing freckle disease, was confirmed. Freckle infection also induced other biochemical defences, like pathogenesis-related proteins, phenolics and other structural defences. A banana leaf bioassay system was developed for assessing resistance-inducing capacity of elicitors derived from the banana freckle and anthracnose pathogens. In field trials, preharvest treatment with Bion and salicylic acid reduced anthracnose and crown rot, and stalk-end rot was also reduced by salicylic acid. Fertiliser field trials demonstrated that increased nitrogen enhances anthracnose, while application of potassium reduced anthracnose and finger-end rot, particularly in soils with low initial levels of potassium. As with mango, cultivar differences in the resistance (or susceptibility) to anthracnose were demonstrated. It is recommended that further field trials focus on the incorporation of Bion into field disease management programs, but its registration and adoption remains the decision of Syngenta. Other defence activators should be assessed as they become available. Postharvest UV-C treatment should be assessed under commercial packing-line conditions, and it is hoped that this will have application to disease management in mango in the near future. The information on nitrogen fertilisation in banana and mango could have immediate impact if made widely available to growers and other agricultural/extension staff. The short-term impact of the variety work is that growers/industry could choose more disease-resistant varieties. In the longer term, the selection and adoption of more resistant rootstocks (mango) is feasible and the work on biochemical defences could lead to assays for screening germplasm for resistance as part of a breeding program. The global knowledge of natural plant defence and what affects it has been significantly enhanced in this project, and the capacity of all project teams to conduct such research has been elevated. Some information is available immediately to industry and has been disseminated via workshops and field days.

Project ID:
HORT/1997/094

Start Date

01/07/2002

Project Coordinator Fax

(02) 6217 0501

Reference Number
BR-972706-35395

Project Type
Bilateral

Project Status
Concluded

Final Progress Report

Tropical and subtropical fruit crops are particularly susceptible to deterioration by postharvest disease, and significant losses are incurred in all production areas. Current control measures rely heavily on the use of fungicides, both before and after harvest. However, for many reasons we can expect that the use of these fungicides will become increasingly limiting. It is thus imperative that alternative or complementary strategies are sought. The project was undertaken to evaluate the prospect of utilising inherent plant defence mechanisms in the management of postharvest diseases focusing on mango (Australia, Sri Lanka and The Philippines) and banana (Sri Lanka). The key diseases were anthracnose in mango and banana, caused by Colletotrichum gloeosporioides and C. musae, respectively, and stem-end rot in mango, caused by Botryosphaeria spp. A significant component was to identify and evaluate activators of plant defences under field conditions. The activators were known resistance-inducing agents, including acibenzolar-S-methyl (Bion), and elicitors derived from fungal pathogens (in banana). Another component was to characterise some of the key biochemical defences contributing to the resistance, and to identify treatments, varietal properties or other agronomic practices which may influence their relative concentrations. The final key objective was to enhance the capacity of project teams to conduct plant defence research, and provide information to respective industries via workshops and field days. Mango was the crop common to project activities in Australia, Sri Lanka and The Philippines. In field trials, Bion was the most consistently effective activator of resistance to anthracnose disease, when applied as a foliar spray or as a soil drench 3-5 times throughout the fruiting period. There is clearly the potential for reducing the number of fungicides applied in a given season if Bion is applied. UV-C treatment to harvested mango activated biochemical defences and reduced anthracnose. Another key finding from field trials in Australia (cv. Keitt) and Sri Lanka (cv. Karuthacolomban) was that increasing nitrogen fertilisation enhances anthracnose levels in fruit, which was correlated with high skin nitrogen and lower levels of preformed alk(en)ylresorcinols in skin tissue. Some mango cultivars (and rootstocks) consistently showed high levels of resistance to anthracnose, eg. 'Keitt' in Australia and 'Gira' and 'Karuthacolomban' in Sri Lanka. There was some correlation between resistance amongst varieties and levels of

constitutive defences. In Sri Lanka the galloyltannin class of compounds was identified to be a major component contributing to antifungal activity in mango peel extracts. The capacity to analyse the alk(en)ylresorcinol compounds was made possible after establishing collaboration with an expert in Poland. All banana work was conducted in Sri Lanka. The existence and partial characterisation of several phenylphenalone-type phytoalexins accumulating in response to infection with Phyllosticta musarum, the pathogen causing freckle disease, was confirmed. Freckle infection also induced other biochemical defences, like PR proteins, phenolics and other structural defences. A banana leaf bioassay system was developed for assessing resistance-inducing capacity of elicitors derived from the banana freckle and anthracnose (Colletotrichum musae) pathogens, and the active components were identified as 5-C polysaccharides (with a protein component for the C. musae elicitor). In field trials, preharvest treatment with Bion and salicylic acid reduced anthracnose and crown rot, and stalk-end rot was also reduced by salicylic acid. Fertiliser field trials demonstrated that increased nitrogen enhances anthracnose, while application of potassium reduced anthracnose and finger-end rot, particularly in soils with low initial levels of potassium. As with mango, cultivar differences in the resistance (or susceptibility) to anthracnose were demonstrated. It is recommended that further field trials focus on the incorporation of Bion into field disease management programs, but its registration and adoption remains the decision of Syngenta. Other defence activators should be assessed as they become available. Postharvest UV-C treatment should be assessed under commercial packingline conditions, and it is hoped that this will have application to disease management in mango in the near future. The information on nitrogen fertilisation in banana and mango could have immediate impact if made widely available to growers and other agricultural/extension staff. The short term impact of the variety work is that growers/industry could choose more disease resistant varieties. In the longer term, the selection and adoption of more resistant rootstocks (mango) is feasible and the work on biochemical defences could lead to the development of assays for screening germplasm for resistance as part of a breeding program. The global knowledge of natural plant defence and what affects it has been significantly enhanced in this project, and the capacity of all project teams to conduct such research has been elevated. Some information is available immediately to industry and has been disseminated via workshops and field days.

Finish Date
31/12/2005

Extension Start Date

01/01/2006

Commissioned Organisation:
Queensland Department of Primary Industries and Fisheries, Australia

dockey
ACIA-6SEW2R

Project Coordinator Email

baxter@aciar.gov.au

Commissioned Organisation
Queensland Department of Primary Industries and Fisheries, Queensland Horticulture Institute, Australia

Extension Finish Date

30/06/2007

Overview Collaborators
Department of Agriculture, Sri Lanka University of Peradeniya, Sri Lanka Philippine Council for Agriculture, Forestry and Natural Resources Research and Development, Philippines University of the Philippines at Los Banos, Philippines

ACIAR Research Program Manager

Mr Les Baxter

Progress Reports (Year 1, 2, 3 etc) Year 1:

Subproject/objective 1. Host defence mechanisms in mango and avocado fruit. >>> Within a five week period from December 2002 to January 2003 mango and avocado peel was extracted with appropriate organic solvents and the crude extracts partially purified by column (flash) chromatography. Many resulting fractions demonstrated antifungal activity as determined by TLC plate bioassay, and were analysed by 1HNMR and ESMS. Some of the active fractions were found to contain combinations of fatty acids (palmitic, oleic, linoleic and linolenic acids) and also B-sitosterol. Another promising bioactive fraction could not be satisfactorily identified. The major antifungal compound known in avocado, (Z,Z)-2-hydroxy-4-oxohenicosa-12, 15-dien-1-yl acetate (or diene) was identified. This work is continuing. In Sri Lanka bioactive compounds in mango are also being characterised. Crude analyses of pulp tissue suggest that two antifungal compounds (yet to be identified) may be present at higher concentrations than in the peel. Another significant result obtained within the first year demonstrated that if mango fruits were de-sapped (stalks cut and sap drained out) soon after harvest, the postharvest anthracnose disease was more severe compared to fruits that were not de-sapped (stalks left on). This implies a major role for sap, and its antifungal components, in the resistance of mango to postharvest disease. This work will be repeated and extended to include investigation of Australian cultivars, and also characterisation of the antifungal agents involved.

Field trials in mango cv. Keitt have demonstrated that postharvest anthracnose disease is more severe following applications of high levels of nitrogenous fertiliser through the fruiting period, corroborating previous work. Future work will determine whether levels of antifungal compounds are affected by the high N treatments, and whether the stage of application of N fertiliser affects postharvest disease. Preliminary field trials have also demonstrated that preharvest treatment of fruit with a known resistance "activator", acibenzolar-S-methyl, may reduce the severity and delay the onset of postharvest anthracnose disease. Mango peel samples from these preliminary experiments have been stored at -80C for future determination of the effect of ASM treatments on levels of antifungal compounds. Subproject/objective 2. Freckle-induced resistance in banana fruit. Peel of immature banana fruits cv. `Embul' did not contain detectable levels of preformed antifungal compounds. However, the fruit peel tissues responded to freckle infection by Phyllosticta musarum, with several induced defences, most notably by accumulation of phytoalexins. Freckle-infected fruits are resistant to anthracnose disease caused by Colletotrichum musae. Infection by C. musae elicits only traces of the same phytoalexins. Further experiments indicate that there are differences in types and relative abundance of preformed antifungal compounds as well as phytoalexins among cultivars. The phytoalexins are being purified prior to their characterisation. Additionally, the pH of the peel in freckle-infected cv Embul is maintained at a higher level and is more resistant to anthracnose than cv. Kolikuttu. Further experiments will be carried out to investigate the significance of increased peel pH following freckle infection, in the resistance of fruits to anthracnose. Subproject/objective 3. Research capacity enhancement A project commencement workshop was held at Indooroopilly 10-14 February 2003. It was attended by all Australian researchers associated with the project (DPI and UQ), and the project leader and three project scientists from Sri Lanka. The workshop was also attended by additional Israeli and Australian scientists with significant expertise in related fields of study. All participants benefited from the interactions and discussions on research plans, methodologies etc. A workshop handbook was produced and is a valuable resource. The Australian project leader visited Sri Lanka in July 2002 to hold discussions with project teams at the University of Peradeniya and Department of Agriculture. An Australian team member visited Sri Lanka in June 2003 and was able to collaborate with the Department of Agriculture team in the mango nutrition trial. He also presented 2 seminars on Australian mango production, breeding and nutrition research, assisted students with sourcing of reference materials and held discussions with staff regarding project activities. The facilities for plant defence research at the University of Peradeniya have been significantly enhanced in the first year by the purchase of capital equipment, ie. an electrophoresis kit and accessories, a stereo microscope and camera, a freeze dryer, ultrasonic homogeniser and pH meter. These items were purchased in Australia by project staff and then consigned to Sri Lanka.

Year 2:
Year 2 (01/07/2003-30/06/2004) 1. Host defence mechanisms in mango and avocado fruit (Sri Lanka and Australia) .>>>

In Sri Lanka, one antifungal fraction of mango peel extract has been tentatively identified as a mixture of galloyl tannins with glycosidic linkages. Galloyl tannins have previously been isolated from other plant species and have been shown to possess antifungal activity. In both Australia and Sri Lanka, differences were found in the resistance of several mango cultivars to anthracnose. Peel and sap samples have been taken from the different cultivars for analysis of antifungal resorcinols. In both countries it was also found that non-de-sapped mango fruit had lower levels of postharvest disease (anthracnose and stem-end rot) than de-sapped fruit, suggesting that sap plays a role in mango disease resistance. Withholding nitrogen fertiliser or delaying application until late in fruit development resulted in lower anthracnose levels in Keitt mango fruit, compared to fruit from trees receiving earlier applications of nitrogen. Peel samples from this trial are currently awaiting analysis for resorcinol levels. Preharvest applications of acibenzolar-S-methyl (Bion) did not significantly reduce anthracnose levels in Kensington Pride mango fruit. However, there was a trend showing less anthracnose in acibenzolar-S-methyl treated fruit compared to control fruit. In preliminary studies, rootstock choice was shown to influence the susceptibility of Kensington Pride mangoes to anthracnose. In avocado, peel samples were taken from Hass fruitlets sampled at several different stages of fruit development. These are to be analysed for antifungal diene levels in the coming months. Attempts will be made to correlate this data with differences in the susceptibility of avocado fruit to infection by C. gloeosporioides (the anthracnose and pepper spot pathogen) at different stages of fruit maturity. 2. Freckle-induced resistance in banana fruit (Sri Lanka). >> Local banana cultivars exhibited differential susceptibility to freckle disease and anthracnose in Sri Lankan studies. The cultivar "Puvalu", which is highly susceptible to freckle, was found to accumulate high levels of phytoalexins following severe infection by the freckle fungus. As a result, this cultivar is highly resistant to anthracnose. The cultivar Seeni, however, which is resistant to both freckle and anthracnose, was found to contain a preformed antifungal substance that was not detected in any other cultivars tested. Chemical analysis of the phytoalexins induced by freckle infection is underway. Freckle infection was also shown to delay the respiratory climacteric peak and ripening of fruit compared to non-freckled fruit, as well as significantly increasing pH of the fruit peel in all commercial cultivars tested. Harvested banana fruit cultivars of Embul, were sprayed with different concentrations of the defence activators salicylic acid (SA) or acibenzolar-S-methyl (Bion), and inoculated 72 hours later with Colletotrichum musae. SA (1000 mg/l) and Bion (400 mg/l) treatments significantly reduced anthracnose development compared with controls. 3. Research capacity enhancement. >>> Three research students from the University of Peradeniya (Sri Lanka) visited DPI&F (Indooroopilly and Mareeba) in January 2004 to undergo research training. One of the QDPIF team visited the University of Peradeniya in June 2004 and provided research methodology training to the Peradeniya group, particularly in pathogen isolation, identification and field trials. A two day workshop was presented at the Postgraduate Institute of Science (University of Peradeniya) on Postharvest handling and disease control in fruit to project collaborators as well as participants from industry and other institutions.

Year 3:
Australia-Sri Lanka component. >> Subproject/objective 1. >> Host defence mechanisms in mango and avocado fruit. >>>

In Australian research, mango peel and sap extracts have been analysed by HPLC for quantification of constitutive antifungal compounds 5-n-heptadecenylresorcinol (C17:1) and 5-n-pentadecylresorcinol (C15:0). The analyses were performed at UQ, Gatton and also by a collaborator in Poland. The analyses confirm that the heptadecenyl resorcinol is most abundant in mango. Pentadecyl resorcinol is also present in lower amounts. The abundance of the compounds varies among cultivars, eg. Kensington Pride>Keitt=Celebration>Kent=R2E2=Nam Doc Mai=Calypso=Honey Gold=Brooks (peel extracts) and Keitt> Kensington Pride=R2E2>Calypso=Celebration> Honey Gold>Nam Doc Mai (sap extracts). These findings are broadly in line with comparative reaction to inoculation with Colletotrichum gloeosporioides, the anthracnose pathogen, ie. from most tolerant or resistant to most susceptible, Keitt>Kensington Pride>Calypso>Honey Gold>R2E2>Nam Doc Mai. Similar varietal responses to anthracnose have been reported in Sri Lankan cultivars: Twelve Sri Lankan mango cultivars were screened for anthracnose development and the initial studies indicated that the cultivars, 'Petti'and 'Walu' were highly susceptible to anthracnose while cvs. 'Gira', 'KC' , 'Peterpassand', and 'Pol' were the most resistant. 'Kohu' 'Malwan', 'Rata', 'Neelum', 'Wal' and 'Seylum' were moderately susceptible. Antifungal activity levels in the cultivars are being evaluated. A second season's field trial in Australia has demonstrated that maintaining 2-3 cm stalks at harvest could reduce the severity of postharvest anthracnose and stem-end rot diseases. Further analyses demonstrated a strong relationship between resorcinol compounds, skin nitrogen and amount of nitrogen fertiliser applied to mango trees. As skin N increases, resorcinol levels decrease. These studies imply a major role for constitutive resorcinol compounds in the defence response of mango to these pathogens. In other Australian field studies, pre-harvest treatment with the plant defence activator acibenzolar-S-methyl (ASM, Bion) again significantly enhanced resistance to postharvest anthracnose in mango. Another product, soluble potassium silicate, which may function similarly to ASM, was also trialled, and shown to enhance resistance to anthracnose. Treatments were effective when applied three times through the fruiting period as a fruit dip, or soil drench. Field spray treatments with salicylic acid also reduced severity of anthracnose. There was evidence from laboratory tests in Australia and Sri Lanka that postharvest exposure of mango fruit to ultra violet-C radiation, or treatment with ASM or salicylic acid (SA) could also decrease the severity of anthracnose. The apparent induction of resistance by these field and lab treatments was often accompanied by increases in activities of pathogenesis-related proteins, chitinase and B-1-3-glucanase. In mango fertiliser trials conducted in Sri Lanka, high potassium levels were shown to reduce anthracnose severity in fruit whereas high nitrogen levels increased anthracnose. Sri Lankan field trials also showed that high potassium levels reduce anthracnose severity in banana fruit. Project staff in Australia have experienced ongoing difficulties in establishing suitable extraction and HPLC techniques for quantifying diene compounds in avocado peel extracts. We have negotiated access to an HPLC machine in the Forestry building at the QDPIF Indooroopilly Research Centre, but due to time constraints and unexpected inconsistencies in the chromatography, it has not yet been possible to establish a reliable HPLC method that would serve for routine analysis of samples. Recently a project staff member has undertaken training in HPLC theory and practice, and we hope to optimise the analyses with assistance from a new technical officer based at Forestry in the near future. Subproject/objective 2. >>> Freckle-induced resistance in banana fruit. >>> In Sri lanka, polyacrylamide gel electrophoresis of extracts from banana infected with the freckle

pathogen, Phyllosticta musarum, showed presence of at least 6 proteins or peptides that were absent or less intense in non-infected banana. Chitinase activity and total phenolic compounds were also enhanced in infected banana. A polysaccharide fraction from pycnidia of the freckle fungus has been partially purified. It is particularly effective at eliciting phytoalexin accumulation and phenolic-associated browning when applied to banana petiole segments. It is being further purified by gel filtration chromatography. Subproject/objective 3. >>> Research capacity enhancement. >>> The Sri Lankan project leader visited project staff at DPI&F, Indooroopilly and Mareeba in October 2004 where project matters were discussed and visits made to the fields and laboratories. Research capacity at Peradeniya was further enhanced through the purchase of an orbital shaker, magnetic stirrer, research microscope and ammonia meter. Project members are now able to routinely analyse and quantify antifungal resorcinol compounds by HPLC technique after receiving some standard compounds and technical assistance from a Polish collaborator. Australia - Philippines component. >>> This project was extended to the Philippines during this reporting period, with co-funding from AusAID for the Philippines-Australia component The first annual report for the Philippines component is not due until 31 December 2005. In August 2004, the Philippines project leader and partners from the University of the Philippines at Los Baos and Mindanao Provincial agencies visited Australia. The team met colleagues at DPI&F Indooroopilly and Nambour, and were briefed on the activities of the Sri Lankan component of the project. They participated in an undergraduate student session lead by the University of Queensland Gatton supervisor of students who are involved in the project. They then held discussions with their Australian counterparts on project extension matters and jointly developed a tentative work plans for the 2004/2005 and 2005/2006 seasons. In the 2004-2005 Australian mango season, preliminary investigations of effects of activators and other products on diseases in mango were made at trial sites in north Queensland and SE Queensland. The following activators were identified for use in the field trials: a silicon-based product, acibenzolar-S-methyl, Pentra bark (R) surfactant (Agrichem product), and a kaolin and copper base product. The efficacy of these activators in spray and/or drench field applications for the control of postharvest rots of mangoes was investigated singly and in combination with some fungicides. This was compared with an industry fungicide standard program that included azoxystrobin. Preliminary results from the DPI&F Ayr Research Station site and on two mango cultivars, Kensington Pride and R2E2, suggests that none of the activators gave satisfactory control on anthracnose and stem end rots, the two main postharvest diseases of concern, when compared with the standard fungicide program. The use of the activators in combination with two fungicide sprays, however, gave good disease control, comparable to results with five sprays in the standard fungicide spray program. At one orchard, two trunk injections with the silicon product significantly decreased the severity of postharvest anthracnose. The above findings were taken into consideration in revising the 2005-2006 work plans for the Philippines and Australia. Other factors that could have impeded on a response from the activators have been identified and will also be investigated during the 2005 season.

Dr Chrys Akem, DPI&F, Ayr (Australian research leader of the Philippine component), visited the Philippines in April 2005. He participated in a project initiation workshop (held in tandem with another new project PHT/2003/071 mango IPM/Supply chain improvement), visited potential field sites with Filipino partners, assisted in development of a revised work-plan and delivered a lecture to the staff of PCARRD, UPLB and the Crop Protection Institute, focusing on the Australian mango industry and current strategies for the management of field and postharvest diseases.

Year 4:
Objective 1. Host defence mechanisms in mango fruit (Sri Lanka, the Philippines and Australia). Field trials to evaluate the efficacy of defence activators for postharvest disease control in mango were conducted in Australia (North and South Queensland) and the Philippines. In North Queensland, KasilR (potassium silicate) applied as a soil drench and BionR (acibenzolarS-methyl) applied as a foliar spray significantly reduced the incidence of stem-end rot and total postharvest disease in 'R2E2' mango fruit. Reductions in anthracnose by these treatments were not statistically significant this season. There were also no significant reductions in postharvest disease levels in 'Kensington Pride' (KP) mangoes by defence activator treatments at both the North and South Queensland field sites, although there was an encouraging trend of reduced anthracnose by two silicon treatments at one field site in south Queensland. Variability in the response of mango to defence activator treatments may be related to timing and method of application, soil type and regularity of flowering/fruit set . In the Philippines where trials were conducted on 'Carabao' mango at two field sites (Site 1: Davao del Sur, relatively dry mango production area and, Site 2: Davao del Norte, relatively wet mango production area), BionR significantly reduced postharvest anthracnose, blossom blight and scab at site 2. Scab was also reduced by BionR at site 1. KasilR and rice hull ash (a mulch containing over 60% silica) reduced anthracnose and blossom blight under certain conditions at site 2. Follow-up trials are planned for Australia, the Philippines and Sri Lanka for the next mango season. In Sri Lanka, desapping mango fruit significantly reduced both anthracnose and stem-end rot in two local cultivars, confirming results from last season. Desapping 'KP' mangoes in Australia however did not reduce disease, which contrasts with results obtained in Australia last year. Fruit in the current season were very mature with low volumes of sap, which may account for the different result. A further trial to investigate the effect of maturity and desapping on postharvest disease in 'KP' mango is planned for the 06/07 mango season in Australia. A selection of Sri Lankan mango cultivars of differing susceptibility to postharvest disease (anthracnose and stem-end rot) were analysed for chitinase activity in sap, sap volumes and galloyl tannin levels in peel. Peel samples have also been extracted for resorcinol analysis. Results obtained to date indicate some correlation between galloyl tannin levels and anthracnose resistance, although one highly susceptible cultivar did have high levels of galloyl tannin. Results of resorcinol analyses will provide more information when they become available. Objective 2. Host defence mechanisms in banana fruit (Sri Lanka). Higher levels of total soluble phenolic acids, free phenolic acids, glycosidically-bound phenolic acids, ester-bound phenolic acids and cell wall-bound phenolic acids were found in freckleinfected banana peel compared to non-freckled peel. PR-proteins (chitinase and -1,

3-glucanase) were also higher in freckled peel. Elicitors were separated from cell wall extracts of Phyllosticta musarum (freckle pathogen) and Colletotrichum musae (anthracnose pathogen) using gel filtration chromatography and resulting fractions were screened using a banana petiole bioassay. Fractions eliciting the strongest defence responses were then identified for further investigation. The elicitor of C. musae was found to be a protein whereas that of P. musarum contained glycoproteins. Further purification and characterisation of these are continuing. Anthracnose development was lower in banana cultivars which had relatively low peel pH (ie. below 5.5) during ripening. In liquid culture C. musae increased pH levels of the growth medium by secreting ammonia. The data suggests that in certain cultivars mechanisms may exist to counter pathogen-induced pH changes. A field trial was conducted to evaluate the defence activators BionR, salicylic acid and K2HPO4. While results are still being analysed, there are promising indications that treatments reduced anthracnose and crown rot. Objective 3. Research capacity enhancement (Sri Lanka and Australia). Two industry training workshops were held in Sri Lanka in October 2005. The 2-day workshops were designed to instruct participants and improve postharvest handling capacity in Sri Lanka. The topics covered included production issues, numerous aspects of postharvest handling, quality and supply chain management. Field visits were also included to see vegetable production and harvest first hand. The first workshop, 'Postharvest Handling of Fruits and Vegetables' was held at the Postgraduate Institute of Science, University of Peradeniya. The target group was researchers, academics, industry personnel and others possessing sufficient background knowledge. The second workshop, 'Postharvest Handling of Cut Flowers and Vegetables' was held in Nuwara Eliya and was attended by growers, sellers, researchers and industry personnel. Both workshops were highly successful and well attended.

Year 5:
Final report received 17/09/2007

Project Background and Objectives

Sri Lanka, the Philippines and Australia are significant producers of tropical fruit with good prospects for market development. However with current control measures, field and postharvest disease losses can affect productivity and hamper market access. The shelf-life of most tropical and subtropical fruit crops is limited by their high susceptibility to postharvest diseases caused by fungi - examples are the diseases anthracnose and stem-end rot - with losses of 20 per cent common. In mango, anthracnose also blights flowers and can cause complete crop loss before harvest, particularly if rain occurs at flowering. Field application of fungicides (e.g. copper compounds, mancozeb), and postharvest treatment with hot water and fungicides, currently form the basis for control of such pathogens, however due to the inadequacy of current options for field and postharvest disease control, alternative strategies need to be developed. Until recently, little attention has been given to the fact that plants have evolved their own powerful defence mechanisms to limit and prevent disease on developing fruit. These include biochemical (e.g. pathogenesis-related proteins, phytoalexins) and physical (e.g. lignification)

barriers to pathogen invasion, and may be constitutive (preformed) or inducible in nature. The chemical defences, involving preformed or induced chemicals, cause infections to remain localised and quiescent (with colonisation restricted). As climacteric fruit ripen, the defence mechanisms begin to break down (antifungal compound levels drop) and disease begins to develop. Some cultivars have naturally higher levels of the constitutive antifungals (for example the cultivar Hass avocado) and so disease development in ripening fruit is delayed, allowing more fruit to be marketed and consumed before disease develops. Furthermore, constitutive mechanisms may be up-regulated (induced) by a range of elicitors, to enhance host defences (and delay disease development).

Collaborating Institutions
Department of Agriculture, Sri Lanka University of Peradeniya, Sri Lanka Philippine Council for Agriculture, Forestry and Natural Resources Research and Development, Philippines University of the Philippines at Los Banos, Philippines

Program Areas
Horticulture [2]

Overview Methodologies
This project characterised natural disease resistance mechanisms in selected commercially important fruit crops from Sri Lanka (mango, banana), and Australia (mango, avocado). Strategies were developed for their enhancement, using defence-boosting treatments and improved grower practice, to reduce field losses and suppress postharvest disease development (Sri Lanka and the Philippines - mango; Australia - mango and avocado). The Sri Lanka-Australia component of the project undertook a more detailed understanding of the nature and composition of natural defence responses of mango, banana and avocado fruit to pathogen invasion during fruit development and postharvest storage. The Sri Lankan and Australian teams sought to define the physicochemical nature of these host defence systems in terms of genotypic (species, variety or cultivar) and phenotypic (growth rate, production environment) influences. Because harvested fruit becomes less able to suppress pathogen invasion as it ripens, relationships between levels of natural defence systems and fruit physiology (maturity and ripening-related changes in composition) were also examined. The impact of cultural practices on defence systems was also studied as part of formulating recommendations for the field management of postharvest diseases. In other studies the research team investigated the physicochemical host defence systems or each of the host-pathogen systems under study, their induction and maintenance for natural defence. As well, the effectiveness of chemical (e.g. salicylic acid) and/or biological (e.g. non-pathogenic mutant) treatments was studied against the background knowledge of genotype and phenotype influences on host defence systems and their interaction with fruit physiology. The success of the initial work on defence boosting treatments for mango (1/7/2002-30/5/2004) in the Sri Lankan-Australian collaboration led to the expansion of this component to the Philippines. The Philippine-Australian component sought to evaluate and demonstrate sustainable alternative approaches to improve control of field and postharvest diseases of mangoes. This

entailed participatory evaluation of superior treatments at farmer field sites and research stations at several locations.

Overview Objectives
The project aimed to improve control options for diseases of tropical fruit, both in the field and postharvest, by exploring the powerful defence mechanisms that plants have evolved to limit and prevent disease on developing fruit. By so doing, the project also aimed to reduce reliance on synthetic fungicides.

Project Budget
$991912.00 Philippines [1]

Grant Report Value

$1091103.00

Grant Report Recipient

Queensland Department of Primary Industries and Fisheries

Grant Report Recipient Post Code

4068

Grant Report Finish Date

30/06/2007

Grant Report Start Date

25/01/2002

Related publications
Postharvest handling of tropical fruit
[3]

Source URL: http://aciar.gov.au/project/HORT/1997/094 Links: [1] http://aciar.gov.au/country/Philippines [2] http://aciar.gov.au/programarea/Horticulture [3] http://aciar.gov.au/publication/pr050