Effect of nitrogen on the skin colour and other quality attributes of ripe ‘Kensington Pride’ mango (Mangifera indica L.) fruit

Near-ripe ‘Kensington Pride’ mango (Mangifera indica L.) fruit with green skin colour generally return lower wholesale and retail prices. Pre-harvest management, especially nitrogen (N) nutrition, appears to be a major causal factor. To obtain an understanding of the extent of the problem in the Burdekin district (dry tropics; the major production area in Australia), green mature ‘Kensington Pride’ mango fruit were harvested from ten orchards and ripened at 20 ± 0.5 O C. Of these orchards, 70% produced fruit with more than 25% of the skin surface area green when ripe. The following year, the effect of N application on skin colour and other quality attributes was investigated on three orchards, one with a high green (HG) skin problem and two with a low green (LG) skin problem. N was applied at pre-flowering and at panicle emergence at the rate of 0,75,150,300 g per tree (soil applied) or 50 g per tree as foliar N for the HG orchard, and 0,150,300,450 g per tree (soil applied) or 50 g per tree (foliar) for the LG orchards. In all orchards the proportion of green colour on the ripe fruit was significantly (P<0.05) higher with soil applications of 150 g N or more per tree. Foliar sprays resulted in a higher proportion of green colour than the highest soil treatment in the HG orchard, but not in the LG orchards. Anthracnose disease severity was significantly (P<0.05) higher with 300 g of N per tree or foliar treatment in the HG orchard, compared with no additional N. Thus, N can reduce mango fruit quality by increasing green colour and anthracnose disease in ripe fruit.

Getting value out of your sensory testing - the difference from control test

Non-parametric difference tests such as triangle and duo-trio tests traditionally are used to establish differences or similarities between products. However they only supply the researcher with partial answers and often further testing is required to establish the nature, size and direction of differences. This paper looks at the advantages of the difference from control (DFC) test (also known as degree of difference test) and discusses appropriate applications of the test. The scope and principle of the test, panel composition and analysis of results are presented with the aid of suitable examples. Two of the major uses of the DFC test are in quality control and shelf-life testing. The role DFC takes in these areas and the use of other tests to complement the testing is discussed. Controls or standards are important in both these areas and the use of standard products, mental and written standards and blind controls are highlighted. The DFC test has applications in products where the duo-trio and triangle tests cannot be used because of the normal heterogeneity of the product. While the DFC test is a simple difference test it can be structured to give the researcher more valuable data and scope to make informed decisions about their product.

A simplified PCR test for early detection of dwarf off-types in micropropagated Cavendish bananas (Musa spp. AAA)

The dwarf somaclonal variant is a major problem affecting micropropagation of the banana cultivar Williams (Musa spp. AAA; subgroup Cavendish). This problem arises from genetic changes that occur during the tissue culture process. Early identification of this problem is difficult and propagators must wait until plants are ex vitro in order to visualise the dwarfism phenotype. In this study, we have improved a SCAR-based molecular diagnostic technique, developed by Damasco et al. [Acta Hortic. 461 (1997) 157], for the early identification of dwarf off-types. We have included a positive internal control in a multiplex PCR and adapted the technique for use with small amounts of fresh in vitro leaf material as PCR template. The control product is a 500 bp fragment from 18S rRNA and is amplified in all tissues irrespective of phenotype. The use of small in vitro leaf material removing the need for genomic DNA extraction.

Comparison of the indirect haemagglutination and gel diffusion test for serotyping Haemophilus parasuis

The aim of this study was to compare the use of indirect haemagglutination (IHA) and gel diffusion (GD) tests for serotyping Haemophilus parasuis by the Kielstein-Rapp-Gabrielson scheme. All 15 serovar reference strains, 72 Australian field isolates, nine Chinese field isolates, and seven isolates from seven experimentally infected pigs were evaluated with both tests. With the IHA test, 14 of the 15 reference strains were correctly serotyped – with serovar 10 failing to give a titre with serovar 10 antiserum. In the GD test, 13 reference strains were correctly serotyped – with antigen from serovars 7 and 8 failing to react with any antiserum. The IHA methodology serotyped a total of 45 of 81 field isolates while the GD methodology serotyped a total of 48 isolates. For 29 isolates, the GD and IHA methods gave discordant results. It was concluded that the IHA is a good additional test for the serotyping of H. parasuis by the KRG scheme if the GD methodology fails to provide a result or shows unusual cross-reactions.

A universal test to determine the integrity of RNA, and its suitability for reverse transcription, in animal tissue laboratory specimens

Degradation of RNA in diagnostic specimens can cause false-negative test results and potential misdiagnosis when tests rely on the detection of specific RNA sequence. Current molecular methods of checking RNA integrity tend to be host species or group specific, necessitating libraries of primers and reaction conditions. The objective here was to develop a universal (multi-species) quality assurance tool for determining the integrity of RNA in animal tissues submitted to a laboratory for analyses. Ribosomal RNA (16S rRNA) transcribed from the mitochondrial 16S rDNA was used as template material for reverse transcription to cDNA and was amplified using polymerase chain reaction (PCR). As mitochondrial DNA has a high level of conservation, the primers used were shown to reverse transcribe and amplify RNA from every animal species tested. Deliberate degradation of rRNA template through temperature abuse of samples resulted in no reverse transcription and amplification. Samples spiked with viruses showed that single-stranded viral RNA and rRNA in the same sample degraded at similar rates, hence reverse transcription and PCR amplification of 16S rRNA could be used as a test of sample integrity and suitability for analysis that required the sample's RNA, including viral RNA. This test will be an invaluable quality assurance tool for determination of RNA integrity from tissue samples, thus avoiding erroneous test results that might occur if degraded target RNA is used unknowingly as template material for reverse transcription and subsequent PCR amplification.

The vaccination-challenge trial: the gold standard test to evaluate the protective efficacy of infectious coryza vaccines

Infectious coryza is an upper respiratory tract disease of chickens with the major impact occurring in multi-age flocks. We investigated the relationship between the level of antibodies, as detected by a haemagglutination-inhibition (HI) assay, in infectious coryza-vaccinated chickens and the protection against challenge in those chickens. In one experiment, chickens given a single dose of either of two infectious coryza vaccines lacked a detectable HI response to vaccination but showed significant levels of protection 11 weeks after vaccination. In contrast, in chickens given two doses of an infectious coryza vaccine and challenged 3 weeks after the second vaccine dose, there was a strong serological response with 36/40 birds having a HI titre of 1/20 or greater. In this trial there was an apparent relationship between titre and subsequent protection, with none of the 32 chickens with a titre of 1/40 or 1/80 showing any clinical signs and only one of the same group yielding the challenge organism on culture. In contrast, three of the four vaccinated chickens with a HI titre less than 1/5 developed the typical clinical signs of coryza and yielded the challenge organism on culture. Overall, our results suggest that HI titres cannot be regarded as a definitive predictor of vaccine efficacy. We suggest that the vaccination-challenge trial is the gold standard for the evaluation of the immune response to infectious coryza vaccines.

Gene expression in the skin of Bos taurus and Bos indicus cattle infested with the cattle tick, Rhipicephalus (Boophilus) microplus

The cattle tick Rhipicephalus microplus (formerly Boophilus microplus) is responsible for severe production losses to the cattle industry worldwide. It has long been known that different breeds of cattle can resist tick infestation to varying degrees; however, the mechanisms by which resistant cattle prevent heavy infestation are largely unknown. The aim of this study was to determine whether gene expression varied significantly between skin sampling sites (neck, chest and tail region), and whether changes in gene expression could be detected in samples taken at tick attachment sites (tick attached to skin sample) compared with samples taken from non-attachment sites (no tick attachment). We present here the results of an experiment examining the expression of a panel of forty-four genes in skin sections taken from Bos indicus (Brahman) cattle of known high resistance, and Bos taurus (Holstein-Friesian) cattle of known low resistance to the cattle tick. The forty-four genes chosen for this study included genes known to be involved in several immune processes, some structural genes, and some genes previously suggested to be of importance in tick resistance by other researchers. The expression of fifteen gene transcripts increased significantly in Holstein-Friesian skin samples at tick attachment sites. The higher expression of many genes involved in innate inflammatory processes in the Holstein-Friesian animals at tick attachment sites suggests this breed is exhibiting a non-directed pathological response to infestation. Of the forty-four genes analysed, no transcripts were detected in higher abundance at tick attachment sites in the Brahman cattle compared with similar samples from the Holstein-Friesian group, nor difference between attachment site and non-attachment site samples within the Brahman group. The results presented here suggest that the means by which these two cattle breeds respond to tick infestation differ and warrant further investigation.

Expression of intracellular calcium signalling genes in cattle skin during tick infestation

It is widely acknowledged that changes in intracellular calcium ion (Ca2+) concentration provide dynamic signals that control a plethora of cellular processes, including triggering and mediating host defence mechanisms. In this study, quantitative real-time PCR was used to analyse gene expression of 14 Ca2+ signalling proteins in skin obtained from high tick-resistant (HR) and low tick-resistant (LR) cattle following artificial challenge with cattle tick (Rhipicephalus (Boophilus) microplus). Up-regulation of numerous genes was observed in both HR and LR skin following tick challenge, however substantially higher transcription activation was found in HR tissue. The elevated expression in HR skin of specific Ca2+ signalling genes such as AHNAK, CASQ, IL2, NFAT2CIP and PLCG1 may be related to host resistance. Our data suggest that Ca2+ and its associated proteins might play an important role in host response to ticks and that further investigation is warranted.

Test of the enemy release hypothesis: the native magpie moth prefers a native fireweed (Senecio pinnatifolius) to its introduced congener (S. madagascariensis).

The enemy release hypothesis predicts that native herbivores will either prefer or cause more damage to native than introduced plant species. We tested this using preference and performance experiments in the laboratory and surveys of leaf damage caused by the magpie moth Nyctemera amica on a co-occuring native and introduced species of fireweed (Senecio) in eastern Australia. In the laboratory, ovipositing females and feeding larvae preferred the native S. pinnatifolius over the introduced S. madagascariensis. Larvae performed equally well on foliage of S. pinnatifolius and S. madagascariensis: pupal weights did not differ between insects reared on the two species, but growth rates were significantly faster on S. pinnatifolius. In the field, foliage damage was significantly greater on native S. pinnatifolius than introduced S. madagascariensis. These results support the enemy release hypothesis, and suggest that the failure of native consumers to switch to introduced species contributes to their invasive success. Both plant species experienced reduced, rather than increased, levels of herbivory when growing in mixed populations, as opposed to pure stands in the field; thus, there was no evidence that apparent competition occurred.

Skin damage to two new mango cultivars during irradiation and cold storage.

Mangoes can express several skin disorders following important postharvest treatments. Responses are often cultivar specific. This paper reports the responses of two new Australian mango cultivars to some of these treatments. 'Honey Gold' mango develops "under skin browning" early during cold storage. This is thought to be partly caused by a discolouration of the latex vessels which then spreads to the surrounding cells. The symptoms appear to be worse in fruit from hotter production areas and that have been cooled to temperatures below 18C soon after harvest. Current commercial recommendations are to cool fruit to 18C, which limits postharvest handling options. Recent trials have confirmed that delayed or slowed cooling after harvest can reduce under skin browning. The defect may also be associated with physical injury to the skin during harvesting and packing. Irradiation is potentially an important disinfestation treatment for fruit fly in Australian mangoes. The 'B74' mango cultivar develops significant skin damage following irradiation, mainly due to discolouration of the cells surrounding the lenticels. Recent results confirmed that fruit harvested directly from the tree into trays without exposure to water or postharvest chemicals are not damaged by irradiation, while commercially harvested and packed fruit are damaged. Several major harvest and postharvest steps appear to increase lenticel sensitivity to irradiation. Further work is required to develop commercially acceptable protocols to facilitate 'Honey Gold' and 'B74' mango distribution and marketing.

Test and develop sustainable and profitable grazing strategies to manage for rainfall variability

The Wambiana grazing trial started in 1997 to test and develop sustainable and profitable grazing strategies to manage for rainfall variability. It is important that this trial continue as the results are still relatively short-term relative to rainfall cycles and significant treatment changes are still occurring. This new proposal will maintain baseline treatments but will modify others based on trial learning’s to date. It builds on treatment differences and evidence collected over the last 12 years to deliver evidence-based guidelines and principles for sustainable and productive management. The trial also links to other projects modelling water quality, climate change, methane emissions and soil C sequestration on grazing lands.

Development of commercial application of an avocado fruit robustness test

Fruit quality is one of the major factors limiting growth in avocado retail sales. Avocado growers are often unaware of their end-use fruit quality since quality problems only manifest upon fruit ripening and growers receive limited feedback from the supply chain. If growers were aware of their expected fruit quality they would be equipped to make better marketing decisions and if necessary to take remedial actions to improve their fruit quality. Avotest is being developed as a quick and easy method of determining expected end-use fruit quality before the start of the commercial fruit harvest. The test aims at distinguishing between blocks with robust fruit and those with less robust fruit. The test could also be used to predict the resulting fruit quality after the implementation of new farming practices.

Reducing skin damage and improving efficiency of Calypso mango

Calypso mango is a relatively new variety owned by DEEDI and managed/marketed by One Harvest (Queensland-based). It is a major mango variety for the retail chains. Its main limitation is a sensitive skin, which results in lenticel spotting and skin browning.

Reducing flesh bruising and skin spotting in Hass avocado

Up to 80% of avocados on the retail shelf have defects in the flesh which reduces consumer satisfaction. Flesh bruising is the single most important contributor. Avocados also develop skin spotting during harvesting and packing which can reduce domestic and international customer confidence. This project will identify where bruising occurs, develop decision aid tools to help industry reduce flesh bruising in ripe fruit, and understand the commercial impacts of skin spotting. The project will include a PhD student with stipend coming from an international scholarship and in kind support from the University of Queensland.

Corymbia leaf oils, latitude, hybrids and herbivory: a test using common-garden field trials

Foliar oils, particularly monoterpenes, can influence the susceptibility of plants to herbivory. In plants, including eucalypts, monoterpenes are often associated with plant defence. A recent analysis revealed an increase in foliar oil content with increasing latitudinal endemism, and we tested this pattern using three eucalypt taxa comprising a latitudinal replacement cline. We also examined the relative concentrations of two monoterpenes (alpha-pinene and 1,8-cineole), for which meta-analyses also showed latitudinal variation, using hybrids of these three taxa with Corymbia torelliana. These, and pure C. torelliana, were then assessed in common-garden field plots for the abundance and distribution of herbivory by four distinct herbivore taxa. Differing feeding strategies among these herbivores allowed us to test hypotheses regarding heritability of susceptibility and relationships to alpha-pinene and 1,8-cineole. We found no support for an increase in foliar oil content with increasing latitude, nor did our analysis support predictions for consistent variation in alpha-pinene and 1,8-cineole contents with latitude. However, herbivore species showed differential responses to different taxa and monoterpene contents. For example, eriophyid mites, the most monophagous of our censused herbivores, avoided the pure species, but fed on hybrid taxa, supporting hypotheses on hybrid susceptibility. The most polyphagous herbivore (leaf blister sawfly Phylacteophaga froggatti) showed no evidence of response to plant secondary metabolites, while the distribution and abundance patterns of Paropsis atomaria showed some relationship to monoterpene yields.