Five commercial citrus rate poorly as hosts of the polyphagous fruit flyBactrocera tryoni(Froggatt) (Diptera: Tephritidae) in laboratory studies

Queensland fruit fly, Bactrocera tryoni (Froggatt), is a polyphagous pest, and many citrus types are included among its hosts. While quantification of citrus host use by B. tryoni is lacking, citrus is generally considered a ‘low pressure’ crop. This paper investigates B. tryoni female oviposition preference and offspring performance in five citrus types; Murcott mandarin (Citrus reticulata), Navel orange and Valencia orange (Citrus sinensis), Eureka lemon (Citrus limon) and yellow grapefruit (Citrus paradisi). Oviposition preference was investigated in laboratory-based choice and no-choice experiments, while immature survival and offspring performance were investigated by infesting fruits in the laboratory and evaluating pupal recovery, pupal emergence and F1 fecundity. Fruit size, Brix level and peel toughness were also measured for correlation with host use. Bactrocera tryoni demonstrated an oviposition preference hierarchy among the citrus fruits tested; Murcott and grapefruit were most preferred for oviposition and lemon the least, while preference for Navel and Valencia was intermediate. Peel toughness was negatively correlated with B. tryoni oviposition preference, while no significant correlations were detected between oviposition and Brix level or fruit size. Immature survival in the tested fruit was very low. Murcott was the best host (21% pupal recovery), while all other citrus types that showed pupal recovery of 6% or lower and no pupae were recovered from Valencia orange. In pupae recovered from Navel orange and lemon, adult eclosion was greatly reduced, while in grapefruit and lemon, no eggs were recovered from F1 adults. Based on these laboratory results, many commercial citrus varieties appear to be poor hosts for B. tryoni and may pose a low post-harvest and quarantine risk. These findings need to be confirmed in the field, as they impact on both pre-harvest and post-harvest countermeasures.

A finite element model of seat cushion indentation with a soft tissue human occupant model

Effective digital human model (DHM) simulation of automotive driver packaging ergonomics, safety and comfort depends on accurate modelling of occupant posture, which is strongly related to the mechanical interaction between human body soft tissue and flexible seat components. This paper presents a finite-element study simulating the deflection of seat cushion foam and supportive seat structures, as well as human buttock and thigh soft tissue when seated. The three-dimensional data used for modelling thigh and buttock geometry were taken on one 95th percentile male subject, representing the bivariate percentiles of the combined hip breadth (seated) and buttock-to-knee length distributions of a selected Australian and US population. A thigh-buttock surface shell based on this data was generated for the analytic model. A 6mm neoprene layer was offset from the shell to account for the compression of body tissue expected through sitting in a seat. The thigh-buttock model is therefore made of two layers, covering thin to moderate thigh and buttock proportions, but not more fleshy sizes. To replicate the effects of skin and fat, the neoprene rubber layer was modelled as a hyperelastic material with viscoelastic behaviour in a Neo-Hookean material model. Finite element (FE) analysis was performed in ANSYS V13 WB (Canonsburg, USA). It is hypothesized that the presented FE simulation delivers a valid result, compared to a standard SAE physical test and the real phenomenon of human-seat indentation. The analytical model is based on the CAD assembly of a Ford Territory seat. The optimized seat frame, suspension and foam pad CAD data were transformed and meshed into FE models and indented by the two layer, soft surface human FE model. Converging results with the least computational effort were achieved for a bonded connection between cushion and seat base as well as cushion and suspension, no separation between neoprene and indenter shell and a frictional connection between cushion pad and neoprene. The result is compared to a previous simulation of an indentation with a hard shell human finite-element model of equal geometry, and to the physical indentation result, which is approached with very high fidelity. We conclude that (a) SAE composite buttock form indentation of a suspended seat cushion can be validly simulated in a FE model of merely similar geometry, but using a two-layer hard/soft structure. (b) Human-seat indentation of a suspended seat cushion can be validly simulated with a simplified human buttock-thigh model for a selected anthropomorphism.

Tensile properties of pumpkin peel and flesh tissue and review of current testing methods

In South and Southeast Asia, postharvest loss causes material waste of up to 66% in fruits and vegetables, 30% in oilseeds and pulses, and 49% in roots and tubers. The efficiency of postharvest equipment directly affects industrial-scale food production. To enhance current processing methods and devices, it is essential to analyze the responses of food materials under loading operations. Food materials undergo different types of mechanical loading during postharvest and processing stages. Therefore, it is important to determine the properties of these materials under different types of loads, such as tensile, compression, and indentation. This study presents a comprehensive analysis of the available literature on the tensile properties of different food samples. The aim of this review was to categorize the available methods of tensile testing for agricultural crops and food materials to investigate an appropriate sample size and tensile test method. The results were then applied to perform tensile tests on pumpkin flesh and peel samples, in particular on arc-sided samples at a constant loading rate of 20 mm min-1. The results showed the maximum tensile stress of pumpkin flesh and peel samples to be 0.535 and 1.45 MPa, respectively. The elastic modulus of the flesh and peel samples was 6.82 and 25.2 MPa, respectively, while the failure modulus values were 14.51 and 30.88 MPa, respectively. The results of the tensile tests were also used to develop a finite element model of mechanical peeling of tough-skinned vegetables. However, to study the effects of deformation rate, moisture content, and texture of the tissue on the tensile responses of food materials, more investigation needs to be done in the future.

Impact of a school-based food garden on attitudes and identification skills regarding vegetables and fruit : a 12-month intervention trial

Objective To determine changes in ability to identify specific vegetables and fruits, and attitudes towards vegetables and fruit, associated with the introduction of a school-based food garden. Design A 12-month intervention trial using a historical control (control n 132, intervention n 120), class-based, self-administered questionnaires requiring one-word answers and 3-point Likert scale responses. Setting A state primary school (grades 4 to 7) in a low socio-economic area of Brisbane, Australia. Intervention The introduction of a school-based food garden, including the funding of a teacher coordinator for 11 h/week to facilitate integration of garden activities into the curriculum. Main outcome measures Ability to identify a series of vegetables and fruits, attitudes towards vegetables and fruit. Analysis Frequency distributions for each item were generated and χ2 analyses were used to determine statistical significance. Exploratory factor analysis was employed to detect major trends in data. Results The intervention led to enhanced ability to identify individual vegetables and fruits, greater attention to origins of produce (garden-grown and fresh), changes to perceived consumption of vegetables and fruits, and enhanced confidence in preparing fruit and vegetable snacks, but decreased interest in trying new fruits. Conclusions The introduction of this school-based food garden was associated with skill and attitudinal changes conducive to enhancing vegetable and fruit consumption. The ways in which such changes might impact on dietary behaviours and intake require further analysis.

Citrus host utilisation by the Queensland fruit fly, Bactrocera tryoni (Frogatt) (Diptera:Tephritidae) : from individuals to populations

Fruit flies are the insects which cause maggots in your backyard fruit and vegetables. They are not just a nuisance to gardeners, but the single greatest insect threat to commercial and subsistence fruit growers throughout Asia, Australia and the Pacific. Queensland fruit fly, the focus of this PhD, costs Australia an estimated $100million per year. I focused specifically on how Queensland fruit fly uses different commercial citrus varieties. I identified specific plant related mechanisms which increase a fruit’s resistance to fruit fly attack. This information can be used by plant breeders to make fruit less prone to fruit fly damage.

Food and Brisbane households : dietary, social and health consequences of food insecurity

Background & Aims: Access to sufficient amounts of safe and culturally-acceptable foods is a fundamental human right. Food security exists when all people, at all times, have physical, social, and economic access to sufficient, safe and nutritious food to meet their dietary needs and food preferences for an active and healthy life. Food insecurity therefore occurs when the availability or access to sufficient amounts of nutritionally-adequate, culturally-appropriate and safe foods, or, the ability to acquire such foods in socially-acceptable ways, is limited. Food insecurity may result in significant adverse effects for the individual and these outcomes may vary between adults and children. Among adults, food insecurity may be associated with overweight or obesity, poorer self-rated general health, depression, increased health-care utilisation and dietary intakes less consistent with national recommendations. Among children, food insecurity may result in poorer self or parent-reported general health, behavioural problems, lower levels of academic achievement and poor social outcomes. The majority of research investigating the potential correlates of food insecurity has been undertaken in the United States (US), where regular national screening for food insecurity is undertaken using a comprehensive multi-item measurement. In Australia, screening for food insecurity takes place on a three yearly basis via the use of a crude, single-item included in the National Health Survey (NHS). This measure has been shown to underestimate the prevalence of food insecurity by 5%. From 1995 – 2004, the prevalence of food insecurity among the Australian population remained stable at 5%. Due to the perceived low prevalence of this issue, screening for food insecurity was not undertaken in the most recent NHS. Furthermore, there are few Australian studies investigating the potential determinants of food insecurity and none investigating potential outcomes among adults and children. This study aimed to examine these issues by a) investigating the prevalence of food insecurity among households residing in disadvantaged urban areas and comparing prevalence rates estimated by the more comprehensive 18-item and 6-item United States Department of Agriculture (USDA) Food Security Survey Module (FSSM) to those estimated by the current single-item measure used for surveillance in Australia and b) investigating the potential determinants and outcomes of food insecurity, Methods: A comprehensive literature review was undertaken to investigate the potential determinants and consequences of food insecurity among developed countries. This was followed by a cross-sectional study in which 1000 households from the most disadvantaged 5% of Brisbane areas were sampled and data collected via mail-based survey (final response rate = 53%, n = 505). Data were collected for food security status, sociodemographic characteristics (household income, education, age, gender, employment status, housing tenure and living arrangements), fruit and vegetable intakes, meat and take-away consumption, presence of depressive symptoms, presence of chronic disease and body mass index (BMI) among adults. Among children, data pertaining to BMI, parent-reported general health, days away from school and activities and behavioural problems were collected. Rasch analysis was used to investigate the psychometric properties of the 18-, 10- and 6-item adaptations of the USDA-FSSM, and McNemar's test was used to investigate the difference in the prevalence of food insecurity as measured by these three adaptations compared to the current single-item measure used in Australia. Chi square and logistic regression were used to investigate the differences in dietary and health outcomes among adults and health and behavioural outcomes among children. Results were adjusted for equivalised household income and, where necessary, for indigenous status, education and family type. Results: Overall, 25% of households in these urbanised-disadvantaged areas reported experiencing food insecurity; this increased to 34% when only households with children were analysed. The current reliance on a single-item measure to screen for food insecurity may underestimate the true burden among the Australian population, as this measure was shown to significantly underestimate the prevalence of food insecurity by five percentage points. Internationally, major potential determinants of food insecurity included poverty and indicators of poverty, such as low-income, unemployment and lower levels of education. Ethnicity, age, transportation and cooking and financial skills were also found to be potential determinants of food insecurity. Among Australian adults in disadvantaged urban areas, food insecurity was associated with a three-fold increase in experiencing poorer self-rated general health and a two-to-five-fold increase in the risk of depression. Furthermore, adults from food insecure households were twoto- three times more likely to have seen a general practitioner and/or been admitted to hospital within the previous six months, compared to their food secure counterparts. Weight status and intakes of fruits, vegetables and meat were not associated with food insecurity. Among Australian households with children, those in the lowest tertile were over 16 times more likely to experience food insecurity compared to those in the highest tertile for income. After adjustment for equivalised household income, children from food insecure households were three times more likely to have missed days away from school or other activities. Furthermore, children from food insecure households displayed a two-fold increase in atypical emotions and behavioural difficulties. Conclusions: Food insecurity is an important public health issue and may contribute to the burden on the health care system through its associations with depression and increased health care utilisation among adults and behavioural and emotional problems among children. Current efforts to monitor food insecurity in Australia do not occur frequently and use a tool that may underestimate the prevalence of food insecurity. Efforts should be made to improve the regularity of screening for food insecurity via the use of a more accurate screening measure. Most of the current strategies that aim to alleviate food insecurity do not sufficiently address the issue of insufficient financial resources for acquiring food; a factor which is an important determinant of food insecurity. Programs to address this issue should be developed in collaboration with groups at higher risk of developing food insecurity and should incorporate strategies to address the issue of low income as a barrier to food acquisition.

Validity of dietary questionnaires in Sri Lankan adults and the association of dietary intake with obesity

Traditionally, infectious diseases and under-nutrition have been considered major health problems in Sri Lanka with little attention paid to obesity and associated non-communicable diseases (NCDs). However, the recent Sri Lanka Diabetes and Cardiovascular Study (SLDCS) reported the epidemic level of obesity, diabetes and metabolic syndrome. Moreover, obesity-associated NCDs is the leading cause of death in Sri Lanka and there is an exponential increase in hospitalization due to NCDs adversely affecting the development of the country. Despite Sri Lanka having a very high prevalence of NCDs and associated mortality, little is known about the causative factors for this burden. It is widely believed that the global NCD epidemic is associated with recent lifestyle changes, especially dietary factors. In the absence of sufficient data on dietary habits in Sri Lanka, successful interventions to manage these serious health issues would not be possible. In view of the current situation the dietary survey was undertaken to assess the intakes of energy, macro-nutrients and selected other nutrients with respect to socio demographic characteristics and the nutritional status of Sri Lankan adults especially focusing on obesity. Another aim of this study was to develop and validate a culturally specific food frequency questionnaire (FFQ) to assess dietary risk factors of NCDs in Sri Lankan adults. Data were collected from a subset of the national SLDCS using a multi-stage, stratified, random sampling procedure (n=500). However, data collection in the SLDCS was affected by the prevailing civil war which resulted in no data being collected from Northern and Eastern provinces. To obtain a nationally representative sample, additional subjects (n=100) were later recruited from the two provinces using similar selection criteria. Ethical Approval for this study was obtained from the Ethical Review Committee, Faculty of Medicine, University of Colombo, Sri Lanka and informed consent was obtained from the subjects before data were collected. Dietary data were obtained using the 24-h Dietary Recall (24HDR) method. Subjects were asked to recall all foods and beverages, consumed over the previous 24-hour period. Respondents were probed for the types of foods and food preparation methods. For the FFQ validation study, a 7-day weight diet record (7-d WDR) was used as the reference method. All foods recorded in the 24 HDR were converted into grams and then intake of energy and nutrients were analysed using NutriSurvey 2007 (EBISpro, Germany) which was modified for Sri Lankan food recipes. Socio-demographic details and body weight perception were collected from interviewer-administrated questionnaire. BMI was calculated and overweight (BMI ≥23 kg.m-2), obesity (BMI ≥25 kg.m-2) and abdominal obesity (Men: WC ≥ 90 cm; Women: WC ≥ 80 cm) were categorized according to Asia-pacific anthropometric cut-offs. The SPSS v. 16 for Windows and Minitab v10 were used for statistical analysis purposes. From a total of 600 eligible subjects, 491 (81.8%) participated of whom 34.5% (n=169) were males. Subjects were well distributed among different socio-economic parameters. A total of 312 different food items were recorded and nutritionists grouped similar food items which resulted in a total of 178 items. After performing step-wise multiple regression, 93 foods explained 90% of the variance for total energy intake, carbohydrates, protein, total fat and dietary fibre. Finally, 90 food items and 12 photographs were selected. Seventy-seven subjects completed (response rate = 65%) the FFQ and 7-day WDR. Estimated mean energy intake (SD) from FFQ (1794±398 kcal) and 7DWR (1698±333 kcal, P<0.001) was significantly different due to a significant overestimation of carbohydrate (~10 g/d, P<0.001) and to some extent fat (~5 g/d, NS). Significant positive correlations were found between the FFQ and 7DWR for energy (r = 0.39), carbohydrate (r = 0.47), protein (r = 0.26), fat (r =0.17) and dietary fiber (r = 0.32). Bland-Altman graphs indicated fairly good agreement between methods with no relationship between bias and average intake of each nutrient examined. The findings from the nutrition survey showed on average, Sri Lankan adults consumed over 14 portions of starch/d; moreover, males consumed 5 more portions of cereal than females. Sri Lankan adults consumed on average 3.56 portions of added sugars/d. Moreover, mean daily intake of fruit (0.43) and vegetable (1.73) portions was well below minimum dietary recommendations (fruits 2 portions/d; vegetables 3 portions/d). The total fruit and vegetable intake was 2.16 portions/d. Daily consumption of meat or alternatives was 1.75 portions and the sum of meat and pulses was 2.78 portions/d. Starchy foods were consumed by all participants and over 88% met the minimum daily recommendations. Importantly, nearly 70% of adults exceeded the maximum daily recommendation for starch (11portions/d) and a considerable proportion consumed larger numbers of starch servings daily, particularly men. More than 12% of men consumed over 25 starch servings/d. In contrast to their starch consumption, participants reported very low intakes of other food groups. Only 11.6%, 2.1% and 3.5% of adults consumed the minimum daily recommended servings of vegetables, fruits, and fruits and vegetables combined, respectively. Six out of ten adult Sri Lankans sampled did not consume any fruits. Milk and dairy consumption was extremely low; over a third of the population did not consume any dairy products and less than 1% of adults consumed 2 portions of dairy/d. A quarter of Sri Lankans did not report consumption of meat and pulses. Regarding protein consumption, 36.2% attained the minimum Sri Lankan recommendation for protein; and significantly more men than women achieved the recommendation of ≥3 servings of meat or alternatives daily (men 42.6%, women 32.8%; P<0.05). Over 70% of energy was derived from carbohydrates (Male:72.8±6.4%, Female:73.9±6.7%), followed by fat (Male:19.9±6.1%, Female:18.5±5.7%) and proteins (Male:10.6±2.1%, Female:10.9±5.6%). The average intake of dietary fiber was 21.3 g/day and 16.3 g/day for males and females, respectively. There was a significant difference in nutritional intake related to ethnicities, areas of residence, education levels and BMI categories. Similarly, dietary diversity was significantly associated with several socio-economic parameters among Sri Lankan adults. Adults with BMI ≥25 kg.m-2 and abdominally obese Sri Lankan adults had the highest diet diversity values. Age-adjusted prevalence (95% confidence interval) of overweight, obesity, and abdominal obesity among Sri Lankan adults were 17.1% (13.8-20.7), 28.8% (24.8-33.1), and 30.8% (26.8-35.2), respectively. Men, compared with women, were less overweight, 14.2% (9.4-20.5) versus 18.5% (14.4-23.3), P = 0.03, less obese, 21.0% (14.9-27.7) versus 32.7% (27.6-38.2), P < .05; and less abdominally obese, 11.9% (7.4-17.8) versus 40.6% (35.1-46.2), P < .05. Although, prevalence of obesity has reached to epidemic level body weight misperception was common among Sri Lankan adults. Two-thirds of overweight males and 44.7% of females considered themselves as in "about right weight". Over one third of both male and female obese subjects perceived themselves as "about right weight" or "underweight". Nearly 32% of centrally obese men and women perceived that their waist circumference is about right. People who perceived overweight or very overweight (n = 154) only 63.6% tried to lose their body weight (n = 98), and quarter of adults seek advices from professionals (n = 39). A number of important conclusions can be drawn from this research project. Firstly, the newly developed FFQ is an acceptable tool for assessing the nutrient intake of Sri Lankans and will assist proper categorization of individuals by dietary exposure. Secondly, a substantial proportion of the Sri Lankan population does not consume a varied and balanced diet, which is suggestive of a close association between the nutrition-related NCDs in the country and unhealthy eating habits. Moreover, dietary diversity is positively associated with several socio-demographic characteristics and obesity among Sri Lankan adults. Lastly, although obesity is a major health issue among Sri Lankan adults, body weight misperception was common among underweight, healthy weight, overweight, and obese adults in Sri Lanka. Over 2/3 of overweight and 1/3 of obese Sri Lankan adults believe that they are in "right weight" or "under-weight" categories.

Child eating behaviour outcomes of an early feeding intervention to reduce risk indicators for child obesity : the NOURISH RCT

Objective Describe parent-reported child eating behaviour and maternal parenting impact outcomes of an infant feeding intervention to reduce child obesity risk. Design and Methods An assessor masked Randomised Controlled Trial (RCT) with concealed allocation of individual mother-infant dyads. The NOURISH RCT enrolled 698 first-time mothers (mean age 30.1 years, SD=5.3) with healthy term infants (51% female) aged 4.3 months (SD=1.0) at baseline. Outcomes were assessed six months post-intervention when the children were 2-years old. Mothers reported on child eating behaviours using the Child Eating Behaviour Questionnaire (CEBQ), food preferences and dietary intake using a 24-hour telephone recall. Parenting was assessed using five scales validated for use in Australia. Results Intervention effects were evident on the CEBQ overall (MANOVA P=.002) and 4/8 subscales: child satiety responsiveness (P=.03), fussiness (P=.01), emotional overeating (P<.01), and food responsiveness (P=.06). Intervention children ‘liked’ more fruits (P<.01) and fewer non-core foods and beverages (Ps=.06, .03). The intervention mothers reported greater ‘autonomy encouragement’ (P=.002) Conclusions Anticipatory guidance on protective feeding practices appears to have modest positive impacts on child eating behaviours that are postulated to reduce future obesity risk.

The potential role of lycopene for the prevention and therapy of prostate cancer : from molecular mechanisms to clinical evidence

Lycopene is a phytochemical that belongs to a group of pigments known as carotenoids. It is red, lipophilic and naturally occurring in many fruits and vegetables, with tomatoes and tomato-based products containing the highest concentrations of bioavailable lycopene. Several epidemiological studies have linked increased lycopene consumption with decreased prostate cancer risk. These findings are supported by in vitro and in vivo experiments showing that lycopene not only enhances the antioxidant response of prostate cells, but that it is even able to inhibit proliferation, induce apoptosis and decrease the metastatic capacity of prostate cancer cells. However, there is still no clearly proven clinical evidence supporting the use of lycopene in the prevention or treatment of prostate cancer, due to the only limited number of published randomized clinical trials and the varying quality of existing studies. The scope of this article is to discuss the potential impact of lycopene on prostate cancer by giving an overview about its molecular mechanisms and clinical effects. © 2013 by the authors; licensee MDPI, Basel, Switzerland.

An R2R3 MYB transcription factor determines red petal colour in an Actinidia (kiwifruit) hybrid population

Background Red colour in kiwifruit results from the presence of anthocyanin pigments. Their expression, however, is complex, and varies among genotypes, species, tissues and environments. An understanding of the biosynthesis, physiology and genetics of the anthocyanins involved, and the control of their expression in different tissues, is required. A complex, the MBW complex, consisting of R2R3-MYB and bHLH transcription factors together with a WD-repeat protein, activates anthocyanin 3-O-galactosyltransferase (F3GT1) to produce anthocyanins. We examined the expression and genetic control of anthocyanins in flowers of Actinidia hybrid families segregating for red and white petal colour. Results Four inter-related backcross families between Actinidia chinensis Planch. var. chinensis and Actinidia eriantha Benth. were identified that segregated 1:1 for red or white petal colour. Flower pigments consisted of five known anthocyanins (two delphinidin-based and three cyanidin-based) and three unknowns. Intensity and hue differed in red petals from pale pink to deep magenta, and while intensity of colour increased with total concentration of anthocyanin, no association was found between any particular anthocyanin data and hue. Real time qPCR demonstrated that an R2R3 MYB, MYB110a, was expressed at significant levels in red-petalled progeny, but not in individuals with white petals. A microsatellite marker was developed that identified alleles that segregated with red petal colour, but not with ovary, stamen filament, or fruit flesh colour in these families. The marker mapped to chromosome 10 in Actinidia. The white petal phenotype was complemented by syringing Agrobacterium tumefaciens carrying Actinidia 35S::MYB110a into the petal tissue. Red pigments developed in white petals both with, and without, co-transformation with Actinidia bHLH partners. MYB110a was shown to directly activate Actinidia F3GT1 in transient assays. Conclusions The transcription factor, MYB110a, regulates anthocyanin production in petals in this hybrid population, but not in other flower tissues or mature fruit. The identification of delphinidin-based anthocyanins in these flowers provides candidates for colour enhancement in novel fruits.

Analysis of genetically modified red-fleshed apples reveals effects on growth and consumer attributes

Consumers of whole foods, such as fruits, demand consistent high quality and seek varieties with enhanced health properties, convenience or novel taste. We have raised the polyphenolic content of apple by genetic engineering of the anthocyanin pathway using the apple transcription factor MYB10. These apples have very high concentrations of foliar, flower and fruit anthocyanins, especially in the fruit peel. Independent lines were examined for impacts on tree growth, photosynthesis and fruit characteristics. Fruit were analysed for changes in metabolite and transcript levels. Fruit were also used in taste trials to study the consumer perception of such a novel apple. No negative taste attributes were associated with the elevated anthocyanins. Modification with this one gene provides near isogenic material and allows us to examine the effects on an established cultivar, with a view to enhancing consumer appeal independently of other fruit qualities. © 2012 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd.

Apple skin patterning is associated with differential expression of MYb10

Background Some apple (Malus × domestica Borkh.) varieties have attractive striping patterns, a quality attribute that is important for determining apple fruit market acceptance. Most apple cultivars (e.g. 'Royal Gala') produce fruit with a defined fruit pigment pattern, but in the case of 'Honeycrisp' apple, trees can produce fruits of two different kinds: striped and blushed. The causes of this phenomenon are unknown. Results Here we show that striped areas of 'Honeycrisp' and 'Royal Gala' are due to sectorial increases in anthocyanin concentration. Transcript levels of the major biosynthetic genes and MYB10, a transcription factor that upregulates apple anthocyanin production, correlated with increased anthocyanin concentration in stripes. However, nucleotide changes in the promoter and coding sequence of MYB10 do not correlate with skin pattern in 'Honeycrisp' and other cultivars differing in peel pigmentation patterns. A survey of methylation levels throughout the coding region of MYB10 and a 2.5 Kb region 5' of the ATG translation start site indicated that an area 900 bp long, starting 1400 bp upstream of the translation start site, is highly methylated. Cytosine methylation was present in all three contexts, with higher methylation levels observed for CHH and CHG (where H is A, C or T) than for CG. Comparisons of methylation levels of the MYB10 promoter in 'Honeycrisp' red and green stripes indicated that they correlate with peel phenotypes, with an enrichment of methylation observed in green stripes. Conclusions Differences in anthocyanin levels between red and green stripes can be explained by differential transcript accumulation of MYB10. Different levels of MYB10 transcript in red versus green stripes are inversely associated with methylation levels in the promoter region. Although observed methylation differences are modest, trends are consistent across years and differences are statistically significant. Methylation may be associated with the presence of a TRIM retrotransposon within the promoter region, but the presence of the TRIM element alone cannot explain the phenotypic variability observed in 'Honeycrisp'. We suggest that methylation in the MYB10 promoter is more variable in 'Honeycrisp' than in 'Royal Gala', leading to more variable color patterns in the peel of this cultivar.

Red colouration in apple fruit is due to the activity of the MYB transcription factor, MdMYB10

Anthocyanin concentration is an important determinant of the colour of many fruits. In apple (Malus x domestica), centuries of breeding have produced numerous varieties in which levels of anthocyanin pigment vary widely and change in response to environmental and developmental stimuli. The apple fruit cortex is usually colourless, although germplasm does exist where the cortex is highly pigmented due to the accumulation of either anthocyanins or carotenoids. From studies in a diverse array of plant species, it is apparent that anthocyanin biosynthesis is controlled at the level of transcription. Here we report the transcript levels of the anthocyanin biosynthetic genes in a red-fleshed apple compared with a white-fleshed cultivar. We also describe an apple MYB transcription factor, MdMYB10, that is similar in sequence to known anthocyanin regulators in other species. We further show that this transcription factor can induce anthocyanin accumulation in both heterologous and homologous systems, generating pigmented patches in transient assays in tobacco leaves and highly pigmented apple plants following stable transformation with constitutively expressed MdMYB10. Efficient induction of anthocyanin biosynthesis in transient assays by MdMYB10 was dependent on the co-expression of two distinct bHLH proteins from apple, MdbHLH3 and MdbHLH33. The strong correlation between the expression of MdMYB10 and apple anthocyanin levels during fruit development suggests that this transcription factor is responsible for controlling anthocyanin biosynthesis in apple fruit; in the red-fleshed cultivar and in the skin of other varieties, there is an induction of MdMYB10 expression concurrent with colour formation during development. Characterization of MdMYB10 has implications for the development of new varieties through classical breeding or a biotechnological approach.

Transcriptional analysis of apple fruit proanthocyanidin biosynthesis

Proanthocyanidins (PAs) are products of the flavonoid pathway, which also leads to the production of anthocyanins and flavonols. Many flavonoids have antioxidant properties and may have beneficial effects for human health. PAs are found in the seeds and fruits of many plants. In apple fruit (Malus × domestica Borkh.), the flavonoid biosynthetic pathway is most active in the skin, with the flavan-3-ols, catechin, and epicatechin acting as the initiating units for the synthesis of PA polymers. This study examined the genes involved in the production of PAs in three apple cultivars: two heritage apple cultivars, Hetlina and Devonshire Quarrenden, and a commercial cultivar, Royal Gala. HPLC analysis shows that tree-ripe fruit from Hetlina and Devonshire Quarrenden had a higher phenolic content than Royal Gala. Epicatechin and catechin biosynthesis is under the control of the biosynthetic enzymes anthocyanidin reductase (ANR) and leucoanthocyanidin reductase (LAR1), respectively. Counter-intuitively, real-time quantitative PCR analysis showed that the expression levels of Royal Gala LAR1 and ANR were significantly higher than those of both Devonshire Quarrenden and Hetlina. This suggests that a compensatory feedback mechanism may be active, whereby low concentrations of PAs may induce higher expression of gene transcripts. Further investigation is required into the regulation of these key enzymes in apple.

Practice and procedure : solicitors' liens for costs

In Patterson v Cohen [2005] NSWSC 635 Hamilton J examined the authorities in relation to what are commonly called 'fruits of litigation' liens. The judgment provides a very useful summary of the principles which apply.