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.

The role of ethylene and cold temperature in the regulation of the apple POLYGALACTURONASE1 gene and fruit softening

Fruit softening in apple (Malus 3 domestica) is associated with an increase in the ripening hormone ethylene. Here, we show that in cv Royal Gala apples that have the ethylene biosynthetic gene ACC OXIDASE1 suppressed, a cold treatment preconditions the apples to soften independently of added ethylene. When a cold treatment is followed by an ethylene treatment, a more rapid softening occurs than in apples that have not had a cold treatment. Apple fruit softening has been associated with the increase in the expression of cell wall hydrolase genes. One such gene, POLYGALACTURONASE1 (PG1), increases in expression both with ethylene and following a cold treatment. Transcriptional regulation of PG1 through the ethylene pathway is likely to be through an ETHYLENE-INSENSITIVE3-like transcription factor, which increases in expression during apple fruit development and transactivates the PG1 promoter in transient assays in the presence of ethylene. A coldrelated gene that resembles a COLD BINDING FACTOR (CBF) class of gene also transactivates the PG1 promoter. The transactivation by the CBF-like gene is greatly enhanced by the addition of exogenous ethylene. These observations give a possible molecular mechanism for the coldand ethylene-regulated control of fruit softening and suggest that either these two pathways act independently and synergistically with each other or cold enhances the ethylene response such that background levels of ethylene in the ethylene-suppressed apples is sufficient to induce fruit softening in apples.

High temperature reduces apple fruit colour via modulation of the anthocyanin regulatory complex

The biosynthesis of anthocyanin in many plants is affected by environmental conditions. In apple (Malus×domestica Borkh.), concentrations of fruit anthocyanins are lower under hot climatic conditions. We examined the anthocyanin accumulation in the peel of maturing 'Mondial Gala' and 'Royal Gala' apples, grown in both temperate and hot climates, and using artificial heating of on-tree fruit. Heat caused a dramatic reduction of both peel anthocyanin concentration and transcripts of the genes of the anthocyanin biosynthetic pathway. Heating fruit rapidly reduced expression of the R2R3 MYB transcription factor (MYB10) responsible for coordinative regulation for red skin colour, as well as expression of other genes in the transcriptional activation complex. A single night of low temperatures is sufficient to elicit a large increase in transcription of MYB10 and consequently the biosynthetic pathway. Candidate genes that can repress anthocyanin biosynthesis did not appear to be responsible for reductions in anthocyanin content. We propose that temperature-induced regulation of anthocyanin biosynthesis is primarily caused by altered transcript levels of the activating anthocyanin regulatory complex.

The genome of the domesticated apple (Malus × domestica Borkh.)

We report a high-quality draft genome sequence of the domesticated apple (Malus × domestica). We show that a relatively recent (>50 million years ago) genome-wide duplication (GWD) has resulted in the transition from nine ancestral chromosomes to 17 chromosomes in the Pyreae. Traces of older GWDs partly support the monophyly of the ancestral paleohexaploidy of eudicots. Phylogenetic reconstruction of Pyreae and the genus Malus, relative to major Rosaceae taxa, identified the progenitor of the cultivated apple as M. sieversii. Expansion of gene families reported to be involved in fruit development may explain formation of the pome, a Pyreae-specific false fruit that develops by proliferation of the basal part of the sepals, the receptacle. In apple, a subclade of MADS-box genes, normally involved in flower and fruit development, is expanded to include 15 members, as are other gene families involved in Rosaceae-specific metabolism, such as transport and assimilation of sorbitol.

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.

Rescaling clustering trees using impact ratios for robust hierarchical speaker clustering

We present a novel method for improving hierarchical speaker clustering in the tasks of speaker diarization and speaker linking. In hierarchical clustering, a tree can be formed that demonstrates various levels of clustering. We propose a ratio that expresses the impact of each cluster on the formation of this tree and use this to rescale cluster scores. This provides score normalisation based on the impact of each cluster. We use a state-of-the-art speaker diarization and linking system across the SAIVT-BNEWS corpus to show that our proposed impact ratio can provide a relative improvement of 16% in diarization error rate (DER).

Effect of cell wall properties on porosity and shrinkage of dried apple

Water removal during drying depends on the pathway of water migration from food materials. Moreover, the water removal rate also depends on the characteristics of the cell wall of plant tissue. In this study, the influence of cell wall properties (such as moisture distribution, stiffness, thickness and cell dimension) on porosity and shrinkage of dried product was investigated. Cell wall stiffness depends on a complex combination of plant cell microstructure, composition of food materials and the water-holding capacity of the cell. In this work, a preliminary investigation of the cell wall properties of apple was conducted in order to predict changes of porosity and shrinkage during drying. Cell wall characteristics of two types of apple (Granny Smith and Red Delicious) were investigated under convective drying to correlate with porosity and shrinkage. A scanning electron microscope (SEM), 2kN Intron, pycnometer and ImageJ software were used in order to measure and analyse cell characteristics, water holding capacity of cell walls, porosity and shrinkage. The cell firmness of the Red Delicious apple was found to be higher than for Granny Smith apples. A remarkable relationship was observed between cell wall characteristics when compare with heat and mass transfer characteristics. It was also found that the evolution of porosity and shrinkage are noticeably influenced by the nature of the cell wall during convective drying. This study has revealed a better understanding of porosity and the shrinkage of dried food at microscopy (cell) level, and will provide better insights to attain energy-effective drying processes and improved quality of dried foods.

Now we can “see the forest and the trees too” but there are risks : Camera trapping and privacy law in Australia

The use of camera traps in wildlife management is an increasingly common practice. A phenomenon which is also becoming more common is for such camera traps to unintentionally film individuals engaged in a variety of activities, ranging from the innocent to the nefarious and including lewd or potentially embarrassing behaviour. It is therefore possible for the use of camera traps to accidentally encroach upon the privacy rights of persons who venture into the area of surveillance. In this chapter we describe the legal framework of privacy in Australia and discuss the potential risk of this sleeping tiger for users of camera traps. We also present the results of a survey of camera trap users to assess the frequency of such unintended captures and the nature of activity being filmed before discussing the practical implications of these laws for camera traps users in this country and make recommendations.

Left ventricular volume and valve gradient analysis using an Apple Macintosh computer

A description of a computer program to analyse cine angiograms of the heart and pressure waveforms to calculate valve gradients.

Using decision trees to understand structure in missing data

Objectives Demonstrate the application of decision trees – classification and regression trees (CARTs), and their cousins, boosted regression trees (BRTs) – to understand structure in missing data. Setting Data taken from employees at three different industry sites in Australia. Participants 7915 observations were included. Materials and Methods The approach was evaluated using an occupational health dataset comprising results of questionnaires, medical tests, and environmental monitoring. Statistical methods included standard statistical tests and the ‘rpart’ and ‘gbm’ packages for CART and BRT analyses, respectively, from the statistical software ‘R’. A simulation study was conducted to explore the capability of decision tree models in describing data with missingness artificially introduced. Results CART and BRT models were effective in highlighting a missingness structure in the data, related to the Type of data (medical or environmental), the site in which it was collected, the number of visits and the presence of extreme values. The simulation study revealed that CART models were able to identify variables and values responsible for inducing missingness. There was greater variation in variable importance for unstructured compared to structured missingness. Discussion Both CART and BRT models were effective in describing structural missingness in data. CART models may be preferred over BRT models for exploratory analysis of missing data, and selecting variables important for predicting missingness. BRT models can show how values of other variables influence missingness, which may prove useful for researchers. Conclusion Researchers are encouraged to use CART and BRT models to explore and understand missing data.

Clash of the titans: Apple, Adobe, and Microsoft under fire at IT pricing inquiry

Today, an Australian parliamentary committee grilled the IT titans - Apple, Adobe, and Microsoft - on price discrimination against Australian consumers. The IT companies were evasive under questioning.

ACE Research Vignette 043: Can academic entrepreneurship research identify and explain the next Apple, Google, or (even) Halfbrick studios?

This series of research vignettes is aimed at sharing current and interesting research findings from our team of international entrepreneurship researchers. This vignette, written by Professor Per Davidsson, reports findings on the extremely skewed distributions of entrepreneurship outcomes and other key variables of interest to entrepreneurship research and practice, as well as what this means for what and how we can learn through academic research.

Failure to launch: The self-regulating Md -MYB10R6 gene from apple is active in flowers but not leaves of Petunia

Coloured foliage due to anthocyanin pigments (bronze/red/black) is an attractive trait that is often lacking in many bedding, ornamental and horticultural plants. Apples (Malus × domestica) containing an allelic variant of the anthocyanin regulator, Md-MYB10R6, are highly pigmented throughout the plant, due to autoregulation by MYB10 upon its own promoter. We investigated whether Md-MYB10R6 from apple is capable of functioning within the heterologous host Petunia hybrida to generate plants with novel pigmentation patterns. The Md-MYB10R6 transgene (MYB10–R6pro:MYB10:MYB10term) activated anthocyanin synthesis when transiently expressed in Antirrhinumroseadorsea petals and petunia leaf discs. Stable transgenic petunias containing Md-MYB10R6 lacked foliar pigmentation but had coloured flowers, complementing the an2 phenotype of ‘Mitchell’ petunia. The absence of foliar pigmentation was due to the failure of the Md-MYB10R6 gene to self-activate in vegetative tissues, suggesting that additional protein partners are required for Md-MYB10 to activate target genes in this heterologous system. In petunia flowers, where endogenous components including MYB-bHLH-WDR (MBW) proteins were present, expression of the Md-MYB10R6 promoter was initiated, allowing auto-regulation to occur and activating anthocyanin production. Md-MYB10 is capable of operating within the petunia MBW gene regulation network that controls the expression of the anthocyanin biosynthesis genes, AN1 (bHLH) and MYBx (R3-MYB repressor) in petals.

Can’t see the trees for the forest? Why IS-ServQual items matter

Despite longstanding concern with the dimensionality of the service quality construct as measured by ServQual and IS-ServQual instruments, variations on the IS-ServQual instrument have been enduringly prominent in both academic research and practice in the field of IS. We explain the continuing popularity of the instrument based on the salience of the item set for predicting overall customer satisfaction, suggesting that the preoccupation with the dimensions has been a distraction. The implicit mutual exclusivity of the items suggests a more appropriate conceptualization of IS-ServQual as a formative index. This conceptualization resolves the paradox in IS-ServQual research, that of how an instrument with such well-known and well-documented weaknesses continue to be very influential and widely used by academics and practitioners. A formative conceptualization acknowledges and addresses the criticisms of IS-ServQual, while simultaneously explaining its enduring salience by focusing on the items rather than the “dimensions.” By employing an opportunistic sample and adopting the most recent IS-ServQual instrument published in a leading IS journal (virtually, any valid IS- ServQual sample in combination with a previously tested instrument variant would suffice for study purposes), we demonstrate that when re-specified as both first-order and second-order formatives, IS-ServQual has good model quality metrics and high predictive power on customer satisfaction. We conclude that this formative specification has higher practical use and is more defensible theoretically.