Progressive Decline in Xylem Inflow into Developing Plums

Recent evidence suggests xylem functionality may decline in developing European plums. Loss of xylem function may have negative consequences for fruit quality. The aim of this study was to establish and localize the loss of xylem functionality, both spatially and temporally using detached fruit. Fruit were detached from the tree under water and fed through a capillary mounted on the cut end of the pedicel. The rate of water movement through the capillary was recorded. Fruit were held above dry silica gel [≈0% relative humidity (RH)] or above water (≈100% RH) to maximize or minimize transpiration, respectively. Water inflow rate depended on developmental stage. It increased from stage I to a maximum at early stage III and then decreased until maturity. Feeding acid fuchsin to developing fruit revealed a progressive decline in dye distribution. The decline progressed basipetally, from the stylar end toward the stem end. At the mature stage III, only the pedicel/fruit junction was stained. The same pattern was observed in four further plum cultivars at the mature stage III. The inflow into early stage III fruit decreased as the RH increased. In contrast, the inflow was less dependent of RH at the mature stage III. Abrading the fruit skin cuticle had no effect on water inflow during early and mature stage III but did markedly increase fruit transpiration rate. Decreasing the osmotic potential (more concentrated) of the feeding solution decreased the water inflow. Our results indicate a progressive loss of xylem functionality in European plum. Transpiration and osmotic pull are the main drivers of this xylem inflow

Low cuticle deposition rate in ‘Apple’ mango increases elastic strain, weakens the cuticle and increases russet

Russeting compromises appearance and downgrades the market value of many fruitcrops, including of the mango cv. ‘Apple’. The objective was to identify the mechanistic basis of ‘Apple’ mango’s high susceptibility to russeting. We focused on fruit growth, cuticle deposition, stress/strain relaxation analysis and the mechanical properties of the cuticle. The non-susceptible mango cv. ‘Tommy Atkins’ served for comparison. Compared with ‘Tommy Atkins’, fruit of ‘Apple’ had a lower mass, a smaller surface area and a lower growth rate. There were little differences between the epidermal and hypodermal cells of ‘Apple’ and ‘Tommy Atkins’ including cell size, cell orientation and cell number. Lenticel density decreased during development, being lower in ‘Apple’ than in ‘Tommy Atkins’. The mean lenticel area increased during development but was consistently greater in ‘Apple’ than in ‘Tommy Atkins’. The deposition rate of the cuticular membrane was initially rapid but later slowed till it matched the area expansion rate, thereafter mass per unit area was effectively constant. The cuticle of ‘Apple’ is thinner than that of ‘Tommy Atkins’. Cumulative strain increased sigmoidally with fruit growth. Strains released stepwise on excision and isolation (εexc+iso), and on wax extraction (εextr) were higher in ‘Apple’ than in ‘Tommy Atkins’. Membrane stiffness increased during development being consistently lower in ‘Apple’ than in ‘Tommy Atkins’. Membrane fracture force (Fmax) was low and constant in developing ‘Apple’ but increased in ‘Tommy Atkin’. Membrane strain at fracture (εmax) decreased linearly during development but was lower in ‘Apple’ than in ‘Tommy Atkins’. Frequency of membrane failure associated with lenticels increased during development and was consistently higher in ‘Apple’ than in ‘Tommy Atkins’. The lower rate of cuticular deposition, the higher strain releases on excision, isolation and wax extraction and the weaker cuticle account for the high russet susceptibility of ‘Apple’ mango

Neck shrivel in European plum is caused by cuticular microcracks, resulting from rapid lateral expansion of the neck late in development

Susceptibility to the commercially important fruit disorder ‘neck shrivel’ differs among European plum cultivars. Radial cuticular microcracking occurs in the neck regions of susceptible cultivars, but not in non-susceptible ones, so would seem to be causal. However, the reason for the microcracking is unknown. The objective was to identify potential relationships between fruit growth pattern and microcracking incidence in the neck (proximal) and stylar (distal) ends of selected shrivel-susceptible and non-susceptible cultivars. Growth analysis revealed two allometric categories: The first category, the ‘narrow-neck’ cultivars, showed hypoallometric growth in the neck region (i.e., slower growth than in the region of maximum diameter) during early development (stages I + II). Later (during stage III) the neck region was ‘filled out’ by hyperallometric growth (i.e., faster than in the region of maximum diameter). The second category, the ‘broad-neck’ cultivars, had more symmetrical, allometric growth (all regions grew equally fast) throughout development. The narrow-neck cultivars exhibited extensive radial cuticular microcracking in the neck region, but little microcracking in the stylar region. In contrast, the broad-neck cultivars exhibited little microcracking overall, with no difference between the neck and stylar regions. Across all cultivars, a positive relationship was obtained for the level of microcracking in the neck region and the difference in allometric growth ratios between stage III and stages I + II. There were no similar relationships for the stylar region. The results demonstrate that accelerated stage III neck growth in the narrow-neck plum cultivars is associated with more microcracking and thus with more shrivel

Russet Susceptibility in Apple Is Associated with Skin Cells that Are Larger, More Variable in Size, and of Reduced Fracture Strain

Russeting is an economically important surface disorder in apple (Malus × domestica Borkh). Indirect evidence suggests an irregular skin structure may be the cause of the phenomenon. The objective of this study was to characterize epidermal and hypodermal cell morphology and the mechanical properties of the skins of apple cultivars of differing russet susceptibility. Dimensions of epidermal and hypodermal cells were determined using microscopy. Stiffness (S), maximum force (Fmax), and maximum strain (εmax) at failure were quantified using uniaxial tensile tests of skin strips. Particularly during early fruit development, epidermal cells (EC) and hypodermal cells (HC) in russet non-susceptible cultivars occurred in greater numbers per unit area than in russet-susceptible ones. The EC and HC were lower in height, shorter in length, and of reduced tangential surface area. There were little differences in S or Fmax between non-susceptible and susceptible cultivars. However, the εmax were higher for the skins of non-susceptible cultivars, than for those of susceptible ones. This difference was larger for the young than for the later growth stages. It is concluded that russet-susceptible cultivars generally have larger cells and a wider distribution of cell sizes for both EC and HC. These result in decreased εmax for the skin during early fruit development when russet susceptibility is high. This increases the chances of skin failures which is known to trigger russeting

Direct Evidence for a Radial Gradient in Age of the Apple Fruit Cuticle

The pattern of cuticle deposition plays an important role in managing strain buildup in fruit cuticles. Cuticular strain is the primary trigger for numerous fruit-surface disorders in many fruit crop species. Recent evidence indicates a strain gradient may exist within the apple fruit cuticle. The outer layers of the cuticle are more strained and thus more susceptible to microcracking than the inner layers. A radial gradient in cuticle age is the most likely explanation. Our study aimed to establish whether (or not) deposition of new cutin in a developing apple fruit occurs on the inner surface of the cuticle, i.e., immediately abutting the outward-facing epidermal cell wall. Developing apples were fed with 13C oleic acid through the skin. Following a 14-d period for incorporation, the fruit was harvested and the cuticular membranes (CMs) isolated enzymatically. The CMs were then ablated to varying extents from the inner or the outer surfaces, using a cold atmospheric pressure plasma (CAPP). Afterwards, the ablated CMs were dewaxed and the 13C contents were determined by mass spectrometry. The incorporation of 13C in the cutin fraction was higher than in the wax fraction. The 13C content was highest in non-ablated, dewaxed CM (DCM) and decreased as ablation depth from the inner surface increased. There was no change in 13C content when ablation was carried out from the outer surface. As fruit development proceeded, more 13C label was found towards the middle of the DCM. These results offered direct evidence for deposition of cutin being on the inner surface of the cuticle, resulting in a radial gradient in cuticular age—the most recent deposition (youngest) being on the inner cuticle surface (abutting the epidermal cell wall) and the earliest deposition (oldest) being on the outer surface (abutting the atmosphere). Copyright © 2021 Si, Khanal, Schlüter and Knoche

Russeting in Apple Is Initiated After Exposure to Moisture Ends—I. Histological Evidence

Russeting (periderm formation) is a critical fruit-surface disorder in apple (Malus × domestica Borkh.). The first symptom of insipient russeting is cuticular microcracking. Humid and rainy weather increases russeting. The aim was to determine the ontogeny of moisture-induced russeting in ‘Pinova’ apple. We recorded the effects of duration of exposure to water and the stage of fruit development at exposure on microcracking, periderm formation and cuticle deposition. Early on (21 or 31 days after full bloom; DAFB) short periods (2 to 12 d) of moisture exposure induced cuticular microcracking—but not later on (66 or 93 DAFB). A periderm was not formed during moisture exposure but 4 d after exposure ended. A periderm was formed in the hypodermis beneath a microcrack. Russeting frequency and severity were low for up to 4 d of moisture exposure but increased after 6 d. Cuticle thickness was not affected by moisture for up to 8 d but decreased for longer exposures. Cuticular ridge thickness decreased around a microcrack. In general, moisture did not affect cuticular strain release. We conclude that a hypodermal periderm forms after termination of moisture exposure and after microcrack formation. Reduced cuticle deposition may cause moisture-induced microcracking and, thus, russeting. © 2020 by the authors. Licensee MDPI, Basel, Switzerland

Time course of changes in the transcriptome during russet induction in apple fruit

Background: Russeting is a major problem in many fruit crops. Russeting is caused by environmental factors such as wounding or moisture exposure of the fruit surface. Despite extensive research, the molecular sequence that triggers russet initiation remains unclear. Here, we present high-resolution transcriptomic data by controlled russet induction at very early stages of fruit development. During Phase I, a patch of the fruit surface is exposed to surface moisture. For Phase II, moisture exposure is terminated, and the formerly exposed surface remains dry. We targeted differentially expressed transcripts as soon as 24 h after russet induction. Results: During moisture exposure (Phase I) of ‘Pinova’ apple, transcripts associated with the cell cycle, cell wall, and cuticle synthesis (SHN3) decrease, while those related to abiotic stress increase. NAC35 and MYB17 were the earliest induced genes during Phase I. They are therefore linked to the initial processes of cuticle microcracking. After moisture removal (Phase II), the expression of genes related to meristematic activity increased (WOX4 within 24 h, MYB84 within 48 h). Genes related to lignin synthesis (MYB52) and suberin synthesis (MYB93, WRKY56) were upregulated within 3 d after moisture removal. WOX4 and AP2B3 are the earliest differentially expressed genes induced in Phase II. They are therefore linked to early events in periderm formation. The expression profiles were consistent between two different seasons and mirrored differences in russet susceptibility in a comparison of cultivars. Furthermore, expression profiles during Phase II of moisture induction were largely identical to those following wounding. Conclusions: The combination of a unique controlled russet induction technique with high-resolution transcriptomic data allowed for the very first time to analyse the formation of cuticular microcracks and periderm in apple fruit immediately after the onset of triggering factors. This data provides valuable insights into the spatial-temporal dynamics of russeting, including the synthesis of cuticles, dedifferentiation of cells, and impregnation of cell walls with suberin and lignin

Apple fruit periderms (russeting) induced by wounding or by moisture have the same histologies, chemistries and gene expressions

Russeting is a cosmetic defect of some fruit skins. Russeting (botanically: induction of periderm formation) can result from various environmental factors including wounding and surface moisture. The objective was to compare periderms resulting from wounding with those from exposure to moisture in developing apple fruit. Wounding or moisture exposure both resulted in cuticular microcracking. Cross-sections revealed suberized hypodermal cell walls by 4 d, and the start of periderm formation by 8 d after wounding or moisture treatment. The expression of selected target genes was similar in wound and moisture induced periderms. Transcription factors involved in the regulation of suberin (MYB93) and lignin (MYB42) synthesis, genes involved in the synthesis (CYP86B1) and the transport (ABCG20) of suberin monomers and two uncharacterized transcription factors (NAC038 and NAC058) were all upregulated in induced periderm samples. Genes involved in cutin (GPAT6, SHN3) and wax synthesis (KCS10, WSD1, CER6) and transport of cutin monomers and wax components (ABCG11) were all downregulated. Levels of typical suberin monomers (ω-hydroxy-C20, -C22 and -C24 acids) and total suberin were high in the periderms, but low in the cuticle. Periderms were induced only when wounding occurred during early fruit development (32 and 66 days after full bloom (DAFB)) but not later (93 DAFB). Wound and moisture induced periderms are very similar morphologically, histologically, compositionally and molecularly

Ultrasonication in Soy Processing for Enhanced Protein and Sugar Yields and Subsequent Bacterial Nisin Production

Soy protein recovery from hexane-defatted soybean flakes using conventional methods is generally low. Importantly, some tightly-bound sugar in the soy flakes ends up in soy protein, thereby deteriorating the usefulness and quality of soy protein as a food ingredient. This research investigated the use of high-power ultrasound prior to soy protein extraction to simultaneously enhance protein yield and facilitate more sugar release in soy whey. The nutrient-rich soy whey was then used as a cheap growth medium to produce high-value nisin using Lactococcus lactis subsp. lactis. A nisin sensitive organism Micrococcus luteus was used as an indicator organism for international unit determination of nisin production as compared to standard. Soy flakes and water was mixed at the ratio of 1:10 (w/w). The slurry was then sonicated for 15, 30, 60 and 120 sec at a frequency of 20 kHz. The ultrasonic amplitude was maintained at 84 µmpp (peak to peak amplitude in µm) for all sonication durations. The results showed that with ultrasound pretreatment, the protein yield improved as much as by 46% in soy extract and sugar release by 50% with respect to nonsonicated samples (control). To maximize nisin production from soy whey, different parameters, such as aeration/agitation and incubation period were optimized. Nisin production from standard medium, DeMan, Rogosa and Sharpe (MRS) and soy whey was tested and compared. Maximum nisin production was achieved in stationary conditions and showed a continuous increase in yield till 48h of incubation (incubation period beyond that was not tested). Maximum nisin yield of 1.78 g/L of soy whey was obtained at 30°C and pH of 4.5 as opposed to 2.96 g/L of nisin with MRS medium

Russeting in apple is initiated after exposure to moisture ends: Molecular and biochemical evidence

Exposure of the fruit surface to moisture during early development is causal in russeting of apple (Malus × domestica Borkh.). Moisture exposure results in formation of microcracks and de-creased cuticle thickness. Periderm differentiation begins in the hypodermis, but only after discon-tinuation of moisture exposure. Expressions of selected genes involved in cutin, wax and suberin synthesis were quantified, as were the wax, cutin and suberin compositions. Experiments were con-ducted in two phases. In Phase I (31 days after full bloom) the fruit surface was exposed to moisture for 6 or 12 d. Phase II was after moisture exposure had been discontinued. Unexposed areas on the same fruit served as unexposed controls. During Phase I, cutin and wax synthesis genes were down-regulated only in the moisture-exposed patches. During Phase II, suberin synthesis genes were up-regulated only in the moisture-exposed patches. The expressions of cutin and wax genes in the moisture-exposed patches increased slightly during Phase II, but the levels of expression were much lower than in the control patches. Amounts and compositions of cutin, wax and suberin were con-sistent with the gene expressions. Thus, moisture-induced russet is a two-step process: moisture exposure reduces cutin and wax synthesis, moisture removal triggers suberin synthesis. © 2020 by the authors. Licensee MDPI, Basel, Switzerland