Developing a commercial hot water treatment to control post-harvest rots on ‘Fiji Red’ papaya

Fiji exports approximately 800 t year-1 of 'Solo Sunrise' papaya marketed as 'Fiji Red' to international markets which include New Zealand, Australia and Japan. The wet weather conditions from November to April each year result in a significant increase in fungal diseases present in Fiji papaya orchards. The two major pathogens that are causing significant post-harvest losses are: stem end rot (Phytophthora palmivora) and anthracnose (Colletotrichum spp.). The high incidence of post-harvest rots has led to increased rejection rates all along the supply chain, causing a reduction in income to farmers, exporters, importers and retailers of Fiji papaya. It has also undermined the superior quality reputation on the market. In response to this issue, the Fiji Papaya industry led by Nature's Way Cooperative, embarked on series of trials supported by the Australian Centre for International Agricultural Research (ACIAR) to determine the most effective and economical post-harvest control in Fiji papaya. Of all the treatments that were examined, a hot water dip treatment was selected by the industry as the most appropriate technology given the level of control that it provide, the cost effectiveness of the treatment and the fact that it was non-chemical. A commercial hot water unit that fits with the existing quarantine treatment and packing facilities has been designed and a cost benefit analysis for the investment carried out. This paper explores the research findings as well as the industry process that has led to the commercial uptake of this important technology.

What influences fungal communities in cotton soils

Soilborne diseases such as Fusarium wilt, Black root rot and Verticillium wilt have significant impact on cotton production. Fungi are an important component of soil biota with capacity to affect pathogen inoculum levels and their disease causing potential. Very little is known about the soil fungal community structure and management effects in Australian cotton soils. We analysed surface soils from ongoing field experiments monitoring cotton performance and disease incidence in three cotton growing regions, collected prior to 2013 planting, for the genetic diversity and abundance as influenced by soil type, environment and management practices and link it with disease incidence and suppression. Results from the 28S LSU rRNA sequencing based analysis indicated a total of 370 fungal genera in all the cotton soils and the top 25 genera in abundance accounted for the major portion of total fungal community. There were significant differences in the composition and genetic diversity of soil fungi between the different field sites from the three cotton growing regions. Results for diversity indices showed significantly greater diversity in the long-term crop rotation experiment at Narrabri (F6E) and experiments at Cowan and Goondiwindi compared to the Biofumigation and D1 field experiments at ACRI, Narrabri. Diversity was lowest in the soils under brassica crop rotation in Biofumigation experiment. Overall, the diversity and abundance of soil fungal community varied significantly in the three cotton growing regions indicating soil type and environmental effects. These results suggest that changes in soil fungal community may play a notable role in soilborne disease incidence in cotton.

The importance of cotton disease surveys in Queensland for monitoring endemic diseases and detecting new pathogens and pests

The cotton industry in Australia funds biannual disease surveys conducted by plant pathologists. The objective of these surveys is to monitor the distribution and importance of key endemic pests and record the presence or absence of new or exotic diseases. Surveys have been conducted in Queensland since 2002/03, with surveillance undertaken by experienced plant pathologists. Monitoring of endemic diseases indicates the impact of farming practices on disease incidence and severity. The information collected gives direction to cotton disease research. Routine diagnostics has provided early detection of new disease problems which include 1) the identification of Nematospora coryli, a pathogenic yeast associated with seed and internal boll rot; and 2) Rotylenchulus reniformis, a plant-parasitic nematode. This finding established the need for an intensive survey of the Theodore district revealing that reniform was prevalent across the district at populations causing up to 30% yield loss. Surveys have identified an exotic defoliating strain (VCG 1A) and non-defoliating strains of Verticillium dahliae, which cause Verticillium wilt. An intensive study of the diversity of V. dahliae and the impact these strains have on cotton are underway. Results demonstrate the necessity of general multi-pest surveillance systems in broad acre agriculture in providing (1) an ongoing evaluation of current integrated disease management practices and (2) early detection for a suite of exotic pests and previously unknown pests.

Rootstock selection, nitrogen and calcium influence postharvest disease in avocado

Field trials evaluating several parameters of growth, fruit yield and quality of 'Hass' avocado grafted to different rootstocks were established in 2004-2005 in four different growing regions of Australia. Fruit were harvested in three seasons from 2008, ripened and assessed for severity and incidence of anthracnose and stem end rot diseases. Peel samples were collected at harvest and analysed for concentrations of the cations (N, K, Ca, Mg). Rootstock significantly affected marketability of fruit (no stem end rot and less than 5% anthracnose) in 58% of the total number of trials evaluated, with better quality fruit harvested from 'Hass' grafted to Guatemalan or West Indian rootstocks such as 'A10' or 'Velvick'. Fruit quality was frequently poor from trees grafted to Mexican race rootstocks, regardless of growing location. Correlation analyses showed that fruit from rootstocks with superior fruit quality was often associated with lower skin N and higher Ca concentrations. There were significant positive correlations between anthracnose and skin N or N:Ca ratio in 75% of trials evaluated. There was a significant negative correlation between anthracnose and Ca in 42% of trials. The correlations between stem end rot and skin N (positive) or Ca (negative) were each significant in 42% of trials. Based on the results in this project, N:Ca ratios in the skin of unripe avocado fruit at harvest may provide one of the best indicators of potential postharvest disease in ripe fruit, and may have implications for fertiliser regimes.

Role of Aeromonas hydrophila in bacterial septicemia of cultured carps in Khouzestan Province and investigation on protectivity of bacterin (s) prepared from acute isolate(s) of it

Motile Aeromonas are the most common bacteria of freshwater in the world that cause disease in fish and other cold-blooded and warm-blooded hosts. Among this group of bacteria, Aeromonas hydrophila is important in causing complications such as fin rot, skin ulcers and lethal hemorrhagic septicemia in fish. Several virulence factors involved in the pathogenesis of Aeromonas hydrophila, including extracellular enzymes (protease, lipase, elastase, gelatinase and nuclease) and toxins. From the exotoxins, hemolysin, aerolysin and cytolytic enterotoxin play an important role in pathogenesis. Detection of virulence markers by PCR as a key component of determining the pathogenesis of the bacteria and using indigenous vaccines for better immunization against this disease is important. In this study, a total of 200 fanned carps (126 common carp. 39 silver carp and 35 of grass carp) with symptoms suspected aeromonas septicemia were isolated from Khouzestan province farms. 125 bacteria belong to Aeromonas genus detected by biochemical and PCR methods. 31 of all isolates recognized as Aeromonas hydrophila with biochemical methods, I6srRNA detection and Lipase genes. Results showed that the role of Aeromonas sp. and Aeromonas hydrophila in fish with disease symptoms were 62.5% and 15.5% respectively. By using specific primers, three virulence genes including hemolysin, aerolysine and cytolytic enterotoxin were detected in these confirmed isolates, that 18 isolates (58/06%) hemolysin positive (hlyA +), 16 isolates (51/61%) aerolysine positive (aerA+) and 23 isolates (74/19%) for cytolytic enterotoxin gene (act+) were positive. The result of present study showed that most of the confirmed isolates genotype was hlyA+ act- with frequency equal to 51/61%. For investigating the protection effect of acut strain of bacteria, UV inactivated bacterin was used.

Termoterapia e quimioterapia de banana 'Prata-Anã' associadas à temperatura de armazenamento no controle de podridões em pós-colheita

O objetivo deste estudo foi avaliar o efeito da termoterapia (56 ºC por 6 min) e quimioterapia (propiconazole 250 ml.l-1) associado com temperatura de armazenamento (temperatura ambiente, 18 ºC e 13 ºC) no controle de podridões de bananas (Musa spp.) 'Prata-Anã' (AAB) em pós-colheita. Os tratamentos apresentaram diferenças significativas na percentagem de área lesionada por fruto, perda de peso e coloração externa da casca em todas as temperaturas de armazenamento. A quimioterapia e a combinação termoterapia e quimioterapia evitaram a manifestação de podridões nas três condições de armazenamento, enquanto a termoterapia reduziu a percentagem de área lesionada por fruto de 98% para 11% em temperatura ambiente, de 8% para 7% em 18 ºC e de 10% para 0% em 13 ºC, sendo mais eficiente sob a temperatura de 13 ºC. Frutos não tratados perderam 25%, 10% e 3% de peso e atingiram a cor 7, 5 e 1 em temperatura ambiente, 18 ºC e 13 ºC, respectivamente. Frutos tratados com termoterapia e quimioterapia perderam 24, 11 e 5% e 20, 10 e 3%, e atingiram índice médio de cor 4 e 3,5, respectivamente. O período de conservação foi estendido para 18, 24 e 45 dias em temperatura ambiente, 18 ºC e 13 ºC, respectivamente. A combinação dos métodos evitou a manifestação de podridões, reduziu a perda de peso e manteve a cor da casca e a qualidade dos frutos.

Termoterapia de banana 'Prata-Anã' no controle de podridões em pós-colheita

O objetivo desse estudo foi determinar a tolerância de banana (Musa spp.) 'Prata-Anã' (AAB) e do fungo Colletotrichum musae à termoterapia no controle de podridões em pós-colheita. Experimentos in vivo e in vitro foram instalados em delineamento inteiramente casualizado, seguindo um esquema fatorial 4x5 (temperatura x tempo). Os tratamentos consistiram na imersão dos frutos (buquês) e do fungo (esporos e micélio) em água aquecida a 47, 50, 53 e 56 ºC, durante 0, 3, 6, 9 e 12 min. A exposição dos frutos a 56 ºC durante 9 min causou escurecimento da casca nas extremidades dos frutos, porém, as características físicas e químicas dos frutos não foram alteradas pelos tratamentos. Frutos inoculados e tratados a 56 ºC durante 6 min não apresentaram podridões nem escurecimento da casca, enquanto aqueles não tratados apresentaram 64% da área lesionada / fruto. A partir das combinações 53 ºC / 9 mi. e 56 ºC / 3 min a germinação de esporos foi reduzida para 4% e 0%, respectivamente. A combinação 56 ºC / 12 min reduziu, mas não paralisou o crescimento micelial. O tratamento 56 ºC / 6 min retardou mas não paralisou o crescimento micelial in vitro, porém foi efetivo no controle completo das podridões in vivo. Esse tratamento evitou a manifestação de podridões no inverno (maio), mas não no verão (novembro), mostrando-se influenciado pelas condições climáticas próximas à colheita dos cachos. A termoterapia pode ser recomendada para controle de podridão em pós-colheita de banana devendo ser ajustada para diferentes estações do ano.

Pesquisa de fungos endofíticos presentes em videira (Vitis vinifera L.) com capacidade para inibir o crescimento do agente causal da podridão negra (Guignardia bidwellii)

Neste trabalho testou-se o potencial antagonista de 16 fungos endofíticos isolados de videiras (Vitis vinifera L.), de castas representativas do Alentejo produzidas em modo de proteção integrada e em modo biológico, contra Guignardia bidwellii. Os isolados identificados após ITS-PCR e sequenciação pertencem aos géneros Epicoccum, Alternaria, Botrytis, Athelia, Phoma e Gibberella. Os isolados testados mostraram atividade antagonista contra G. bidwellii quer por inibição direta, quer através da produção de compostos voláteis, à exceção dos dois isolados de B. cinerea. No entanto, todos os isolados produziram alguns compostos voláteis com reconhecida atividade antimicrobiana, tais como benzaldeído, 3-metil-1-butanol e derivados de ácido propanoico. Foi ainda observado que seis dos isolados produziram também metabolitos não voláteis com capacidade de inibir o crescimento de G. bidwellii. Os resultados obtidos vêm mostrar o potencial dos fungos endofíticos como agentes de luta biológica no controlo de G. bidwellii, podendo constituir novas alternativas no âmbito de Proteção de Plantas; ABSTRACT: Endophytic fungi present in grapevines (Vitis vinifera L.) with the ability to inhibit the growth of the causal agent of black rot (Guignardia bidwellii) In this work the antagonistic potential of 16 endophytic grapevine fungi isolates (Vitis vinifera L.), from representative cultivars of the Alentejo region produced either under integrated pest management or organic mode, was tested against Guignardia bidwellii. Isolates were identified through ITS-PCR and sequencing, as belonging to the genera Epicoccum, Alternaria, Botrytis, Athelia, Phoma and Gibberella. Isolates showed antagonist activity against G. bidwellii either by direct inhibition or through the production of volatile compounds, with the exception of two isolates of B. cinerea. Nevertheless, all isolates produced volatile compounds with known antimicrobial activity such as benzaldehyde, 3-methyl-1-butanol and propionic acid derivatives. Additionally, six isolates produced non-volatile metabolites with the ability to inhibit G. bidwellii growth. These results show the potential that endophytic fungi have as agents for biological control of G. bidwellii, opening new options in the field of Plant Protection.

Effect of pre-bloom leaf defoliation on cluster morphology and disease pressure

Mestrado Vinifera Euromaster - Instituto Superior de Agronomia - UL

Metodos de irrigacao e adubacao na cultura do tomate industrial.

Estudou-se a influencia de dois metodos de irrigacao, dois metodos e duas formulas de adubacao, na cultura do tomate industrial (Lycopersicon esculentum Mill), cultivar Rossol (VFN) usando-se "faixas sub-sub-divididas" em oito repeticoes. Por ocasiao da colheita, foi feita a classificacao, contagem e pesagem dos frutos comerciaveis, verificaram-se diferencas significativas para metodos de adubacao e sua interacao com metodos de irrigacao, ao nivel de probabilidade de 5 e 1%, respectivamente. O metodo de adubacao em sulco apresentou maior rendimento que em cova, quanto irrigado por aspersao, ocorrendo o contrario quando a irrigacao foi por sulco. A producao de frutos com podridao apical foi afetada pelos metodos de irrigacao, metodos e formulas de adubacao, e interacoes, tendo sido encontradas diferencas significativas a niveis de 0,05% de probabilidade. No metodo de irrigacao por aspersao, ocorreu a menor percentagem de frutos com este disturbio fisiologico.

Controle da podridão de raiz e promoção de crescimento em hidroponia com bactérias.

2009

STUDIES OF FUNCTIONALIZED NANOPARTICLES FOR SMART SELF-ASSEMBLY AND AS CONTROLLED DRUG DELIVERY

This dissertation is related to the studies of functionalized nanoparticles for self-assembly and as controlled drug delivery system. The whole topic is composed of two parts. In the first part, the research was conducted to design and synthesize a new type of ionic peptide-functionalized copolymer conjugates for self-assembly into nanoparticle fibers and 3D scaffolds with the ability of multi-drug loading and governing the release rate of each drug for tissue engineering. The self-assembly study confirmed that such peptide-functionalized amphiphilic copolymers underwent different self-assembly behavior. The bigger nanoparticles were more easily assembled into nanoparticle fibers and 3D scaffolds with larger pore size, while the smaller nanoparticle underwent faster self-assembly to form more compact 3D scaffolds with smaller porosity but more stable structure. Controlled release studies confirmed the ability of governing simultaneous release of different model drugs with independent release rate from a same scaffold. Cytotoxicity tests showed that all synthesized peptides, copolymers and peptide-copolymer conjugates were biocompatible with SW-620 cell lines and NIH3T3 cell lines. This new type of self-assembled scaffolds combined the advantages of peptide nanofibers and versatile controlled release of polymeric nanoparticles to achieve simultaneous multi-drug loading and controlled release of each drug, uniform distribution and flexibility of hydrogel scaffolds. The investigations in second part were first to design and synthesize organic biocide-loaded nanoparticles for low-leaching wood preservation using a cost-effective one-pot method to synthesize amphiphilic chitosan-g-PMMA nanoparticles loading with ~25-28 wt.% of the fungicide tebuconazole with particle size of ~100 nm diameter by FESEM. FESEM analysis confirmed efficient penetration of nanoparticles throughout the treated wooden stake with dimension of 19 × 19 × 455 mm^3. Leaching studies showed that biocide introduced into sapwood via nanoparticles leached only ~9% compared with the amount leached from tebuconazole solution-treated control, while soil jar tests showed that the nanoparticle-treated wood blocks were effectively protected from biological decay tested against G. trabeum, a brown rot fungus. Copper oxide nanoparticles with and without polymer stabilizers were also investigated to use as inorganic wood preservatives to clarify the factor affecting copper leaching from treated wood. Copper oxide nanoparticles with uniform diameters of ~10 nm and ~50 nm were prepared, and the leachates from southern pine sapwood treated with these nanoparticles were analyzed. It was found by TEM and EDS analysis that significant numbers of nanoparticles leached from the treated wood. The 50 nm nanoparticles leached slightly less than a soluble copper salt control, but 10 nm nanoparticles leached substantially more than the control. The effect of polymer stabilizers on nanoparticle leaching was also investigated. Results showed that polymer stabilizers increased leaching. The trends showed that nanoparticle size was a major factor in copper leaching.

INTERACTIVE EFFECTS OF CLIMATE CHANGE AND FUNGAL COMMUNITIES ON THE DECOMPOSITION OF WOOD-DERIVED CARBON IN FOREST SOILS

Soils are the largest sinks of carbon in terrestrial ecosystems. Soil organic carbon is important for ecosystem balance as it supplies plants with nutrients, maintains soil structure, and helps control the exchange of CO2 with the atmosphere. The processes in which wood carbon is stabilized and destabilized in forest soils is still not understood completely. This study attempts to measure early wood decomposition by different fungal communities (inoculation with pure colonies of brown or white rot, or the original microbial community) under various interacting treatments: wood quality (wood from +CO2, +CO2+O3, or ambient atmosphere Aspen-FACE treatments from Rhinelander, WI), temperature (ambient or warmed), soil texture (loamy or sandy textured soil), and wood location (plot surface or buried 15cm below surface). Control plots with no wood chips added were also monitored throughout the study. By using isotopically-labelled wood chips from the Aspen-FACE experiment, we are able to track wood-derived carbon losses as soil CO2 efflux and as leached dissolved organic carbon (DOC). We analyzed soil water for chemical characteristics such as, total phenolics, SUVA254, humification, and molecular size. Wood chip samples were also analyzed for their proportion of lignin:carbohydrates using FTIR analysis at three time intervals throughout 12 months of decomposition. After two years of measurements, the average total soil CO2 efflux rates were significantly different depending on wood location, temperature, and wood quality. The wood-derived portion soil CO2 efflux also varied significantly by wood location, temperature, and wood quality. The average total DOC and the wood-derived portion of DOC differed between inoculation treatments, wood location, and temperature. Soil water chemical characteristics varied significantly by inoculation treatments, temperature, and wood quality. After 12 months of decomposition the proportion of lignin:carbohydrates varied significantly by inoculation treatment, with white rot having the only average proportional decrease in lignin:carbohydrates. Both soil CO2 efflux and DOC losses indicate that wood location is important. Carbon losses were greater from surface wood chips compared with buried wood chips, implying the importance of buried wood for total ecosystem carbon stabilization. Treatments associated with climate change also had an effect on the level of decomposition. DOC losses, soil water characteristics, and FTIR data demonstrate the importance of fungal community on the degree of decomposition and the resulting byproducts found throughout the soil.

Exploring the potential of novel biomixtures and Lentinula edodes fungus for the degradation of selected pesticides. Evaluation for use in biobed systems

An approach to reduce the contamination of water sourceswith pesticides is the use of biopurificaction systems. The active core of these systems is the biomixture. The composition of biomixtures depends on the availability of local agro-industrial wastes and design should be adapted to every region. In Portugal, cork processing is generally regarded as environmentally friendly and would be interesting to find applications for its industry residues. In this work the potential use of different substrates in biomixtures, as cork (CBX); cork and straw, coat pine and LECA (Light Expanded Clay Aggregates), was tested on the degradation of terbuthylazine, difenoconazole, diflufenican and pendimethalin pesticides. Bioaugmentation strategies using the white-rot fungus Lentinula edodes inoculated into the CBX, was also assessed. The results obtained from this study clearly demonstrated the relevance of using natural biosorbents as cork residues to increase the capacity of pesticide dissipation in biomixtures for establishing biobeds. Furthermore, higher degradation of all the pesticides was achieved by use of bioaugmented biomixtures. Indeed, the biomixtures inoculated with L. edodes EL1were able to mineralize the selected xenobiotics, revelling that these white-rot fungi might be a suitable fungus for being used as inoculum sources in on-farm sustainable biopurification system, in order to increase its degradation efficiency. After 120 days, maximum degradation of terbuthylazine, difenoconazole, diflufenican and pendimethalin, of bioaugmented CBX, was 89.9%, 75.0%, 65.0% and 99.4%, respectively. The dominant metabolic route of terbuthylazine in biomixtures inoculated with L. edodes EL1 proceeded mainly via hydroxylation, towards production of terbuthylazine-hydroxy-2 metabolite. Finally, sorption process to cork by pesticides proved to be a reversible process,working cork as a mitigating factor reducing the toxicity to microorganisms in the biomixture, especially in the early stages.

Bunch yield of interspecific hybrids of American oil palm with oil palm in the juvenile phase.

To identify superior genotypes, 42 progenies of interspecific hybrids between American oil palm and oil palm were evaluated in an area with incidence of bud rot. The following variables were assessed: total bunch yield (TBY), number of bunches (NB) and mean bunch weight (MBW) from the third to the sixth year of cultivation and estimates of genetic parameters obtained by REML/BLUP procedures.