Profiling of phytohormones in apple fruit and buds regarding their role as potential regulators of flower bud formation.

Apple (Malus × domestica Borkh.) cropping behavior, if not regulated, is often manifested by high yields of small-sized fruit in so called ON-years, which are usually followed by strongly reduced crop loads in OFF-years. Such cropping pattern is defined as biennial bearing and causes significant losses in apple production. The growth of apple fruit overlaps with the formation of flower buds, which remain dormant until the following spring. Earlier works proposed that some fruit-derived mobile compounds, as e.g., phytohormones, could suppress flower bud formation that thereby leads to biennial bearing. We addressed this hypothesis by analyzing 39 phytohormones in apple seeds, fruit flesh and by measuring phytohormone export from the fruits of the biennial bearing cultivar 'Fuji' and of the regular bearing cultivar 'Gala'. Moreover, we analyzed the same compounds in bourse buds from fruiting (ON-trees) and non-fruiting (OFF-trees) spurs of both apple cultivars over the period of flower bud formation. Our results showed that apple fruit exported at least 14 phytohormones including indole-3-acetic acid and gibberellin A3; however, their influence on flower bud formation was inconclusive. A gibberellin-like compound, which was detected exclusively in bourse buds, was significantly more abundant in bourse buds from ON-trees compared with OFF-trees. Cultivar differences were marked by the accumulation of trans-zeatin-O-glucoside in bourse buds of 'Gala' ON-trees, whereas the levels of this compound in 'Gala' OFF were significantly lower and comparable to those in 'Fuji' ON- and OFF-trees. Particular phytohormones including five cytokinin forms as well as abscisic acid and its degradation products had higher levels in bourse buds from OFF-trees compared with ON-trees and were therefore proposed as potential promotors of flower bud initiation. The work discusses regulatory roles of phytohormones in flower bud formation in apple based on the novel and to date most comprehensive phytohormone profiles of apple fruit and buds.

Apple fruit recovery from anoxia under controlled atmosphere storage.

This study investigated the dynamic changes in the anaerobic metabolism end products (ethanol, acetaldehyde and ethyl acetate) of different apple cultivars during 84-days of regular air (RA) and controlled atmosphere (CA) storage after 0-, 3-, 7- and 14-days of anoxia. Increasing the length of exposure to anoxia increased the anaerobic metabolites concentrations (AMC) in all cultivars. Fruit could recover from anoxia, and the AMC dissipated after 84-days of storage under both storage systems when the duration of anoxia was <7-days. The rate of metabolite dissipation following 14-days of anoxia differed among cultivars and storage systems. The reduction in AMC was faster under CA than RA-storage for 'Golden Delicious' and 'Jonagold' fruit, while the opposite trend was observed for 'Kanzi'. These results indicate that storage under CA post anoxia maintains better fruit quality during 84-days of storage than RA and effectively reduces the AMC.

Proteomic differences in apple spur buds from high and non-cropping trees during floral initiation.

The cropping behavior of biennial apple (Malus ×domestica Borkh.) cultivars is irregular and often follows a biennial bearing pattern with 'On' years (high crop load and inhibited floral bud formation) followed by 'Off' years (little crop load and a promoted formation of floral buds). To study proteomic differences between floral and vegetative buds, trees of the strongly alternating cultivar 'Fuji' and the regular bearing cultivar 'Gala' were either completely thinned or not thinned at full bloom to establish two cropping treatments with no ('Off') or a high ('On') crop load, respectively. Student's t-Tests indicated significant differences of protein profiles in buds from 2-year old spurs from both treatments at each sampling date. Abundance patterns of protein clusters coincided with the onset of floral bud initiation and were most noticeable in buds from 'On' trees with a decreased abundance of key enzymes of the phenylpropanoid and flavonoid pathways and an increased abundance of histone deacetylase and ferritins. Furthermore, an increased abundance of proteins involved in histone and DNA methylation was found in the buds from 'Off' trees. This study presents the first large-scale, label-free proteomic profiling of floral and vegetative apple buds during the period of floral bud initiation. SIGNIFICANCE: Although several studies exist that address the complex developmental processes associated with the formation of floral buds in apple (Malus ×domestica Borkh.) at transcriptomic level, no data is available for explaining the difference between floral and vegetative buds or biennial and regular bearing cultivars on a proteomic level. This study presents the first large-scale, label-free proteomic profiling of floral and vegetative apple buds from the two cultivars 'Fuji' and 'Royal Gala' during the period of floral bud initiation and renders possible the development of suitable biomarkers for biennial bearing in apple.

Toward Systematic Understanding of Flower Bud Induction in Apple: A Multi-Omics Approach.

The induction of flower buds in apple (Malus × domestica Borkh.) is tightly connected to biennial bearing, which is characterized by alternating years with high (ON) and low or no (OFF) crop loads. In order to study this irregular cropping behavior, spur buds from ON- and OFF-trees of the biennial-bearing cultivar 'Fuji' and the regular bearing cultivar 'Gala' were collected. First, the time of flower bud initiation was precisely determined for both cultivars by histological analysis. Moreover, for a systematic understanding of flower bud induction in apple, the physiological and molecular mechanisms within the bud tissue were evaluated over four weeks prior to flower bud initiation by employing a multi-omics approach, including RNA sequencing, proteomic and metabolic profiling. Gene and protein enrichment analysis detected physiological pathways promoting and inhibiting early flower bud development. Metabolic profiles from the cropping treatments revealed a greater abundance of thiamine, chlorogenic acid, and an adenine derivative in spur buds from OFF-trees, whereas tryptophan was more abundant in the buds collected from ON-trees. Cultivar comparison indicated that chlorogenic acid was more abundant in 'Gala' than in 'Fuji' spur buds, whereas the opposite effect was found for tryptophan. Genes controlling tryptophan biosynthesis were not affected by ON- and OFF-treatments, but genes assigned to the metabolism of tryptophan into indoleacetate were differentially expressed between cultivars and treatments. The multi-omics approach permitted analyzing complex plant metabolic processes involved in early flower bud development and more specifically presumably in flower bud induction by tracing some pathways from gene to product level.

Climate Change Enhanced Carotenoid Pro-Vitamin A Levels of Selected Plantain Cultivars.

Diet diversification and the exploitation of traditional, micronutrient-rich germplasm of staple crops are generally regarded as sustainable and low-cost approaches to increase the micronutrient intake of resource-poor people. Sun's UV index was collected daily throughout the year. The study assessed the seasonality of provitamin A carotenoids in three plantain cultivars in response to climatic condition. Fruits were harvested at three maturities and freeze-dried before analysis. The results showed that there were high levels of the sun's UV-B radiations throughout the year with the highest occurring from November to May when the area experienced clear skies with minimal cloud cover. These high levels of the sun's UV-B index occurred between 9.00 h GMT and 17.00 h GMT. The study also showed that α-carotene content increased with maturity in "Apantu" during the rainy seasons ranging from 95 to 172 µg/100 g of dry pulp. Similar trends were observed during the dry season with a range of 28 to 489 µg/100 g. The α-carotene contents were very high in the periods of high sun's UV-B radiations compared to the periods of low sun's UV-B radiations. The α-carotene levels in the giant French plantains showed similar trends. Intermediate French "Oniaba" and False Horn "Apantu" plantain cultivar showed the highest content of ß-carotene during the dry season. The high provitamin A carotenoid levels in the cultivars coincided with the high levels of the sun's UV index.

High crop load and low temperature delay the onset of bud initiation in apple.

The reproductive cycle of apple (Malus × domestica Borkh.) starts with the induction of floral development, however, first morphological changes within the bud appear during the following period of bud initiation. This study identifies the onset and duration of bud initiation in the apple cultivars 'Fuji' and 'Gala', characterized by biennial and non-biennial bearing behaviour, respectively, and describes the effect of crop load and heat accumulation on the temporal pattern of floral development. The onset of flower bud initiation in heavy cropping 'Gala' trees was delayed for 20 days compared to trees with no crop load, but the rate of initiation was not affected by crop load. Bud initiation on heavy cropping 'Fuji' trees was minor, whereas trees with no crop load started initiating buds 19 days earlier than those of 'Gala' despite the same cropping status and growing degree hours in a given year. The onset of bud initiation in 'Fuji' 'off' trees was 5 and 20 days after summer solstice, respectively, in two consecutive growing seasons, suggesting that this process is driven by heat accumulation rather than by daylength. The results indicate, that the genetic make-up of the cultivar determines the onset of bud initiation. This can be delayed by increasing crop loads and low temperatures at the beginning of the flower formation process.

Dynamic controlled atmosphere: Impact of elevated storage temperature on anaerobic metabolism and quality of 'Nicoter' apples.

The aim of the study was to evaluate an elevated (3.0 °C) and low (1.0 °C) storage temperature combined with dynamic controlled atmosphere monitored by respiratory quotient (DCA-RQ) and chlorophyll fluorescence (DCA-CF) on anaerobic metabolism, physiological storage disorders and overall quality of 'Nicoter' ('Kanzi®') apples after 5.5 and 8.0 months of storage plus 7d shelf-life. Fruit stored under DCA-RQ 2.0 accumulated the highest amounts of anaerobic metabolites (acetaldehyde, ethanol and ethyl acetate), regardless of storage temperature and timing of storage outturn evaluation, but it did not result in higher electrolyte leakage. Flesh breakdown, core breakdown and cavity formation were reduced at 3 °C. Storage at 3 °C combined with DCA maintained higher flesh firmness after 8.0 months storage plus 7d shelf-life. 'Nicoter' apples can be stored at 3 °C using a DCA system, based either on CF or on RQ, to save electrical energy.

Morphological and biochemical changes during somatic embryogenesis in mahogany, Swietenia macrophylla (Meliaceae)

Intensive exploitation of mahogany wood (Swietenia macrophylla, Meliaceae) has resulted in the loss of natural populations. Somatic embryogenesis offers an alternative to clonal propagation and conservation of mahogany. This study describes biochemical (carbohydrates, total phenols, total flavonoids, protein, and plant growth regulators content) and histological characteristics of the somatic embryogenesis process in mahogany. Calli were obtained by culturing cotyledons of seeds from immature fruits for six weeks on semi-solid MS medium supplemented with 1.0 mgL-1 of kinetin and 4.0 mgL-1 of 2, 4-D. Primary callus was cultured on half strength semi-solid MS medium supplemented with 1.0 mgl-1 6-BA (6-benzylaminopurine) and embryogenic structures were obtained. Embryo development from globular-shaped somatic embryos to the cotyledonary stage was confirmed by histology and scanning electron microscopy. Shoot initiation was observed after somatic embryos were transferred to germination and maturation medium. Endogenous concentrations of carbohydrates, total phenols, total flavonoids, protein, and plant growth regulators were determined in embryogenic (EC) and non-embryogenic (NEC) calli of mahogany. Embryogenic cultures contained significantly higher concentrations of IAA (indoleacetic acid), ABA (abscisic acid), and GAs (Gibberellins 1+3+20), whereas non-embryogenic calli contained more total phenols, flavonoids and resistant starch. Fructose and glucose were not present at detectable levels in EC or NEC, whereas soluble starch and sucrose were only detectable in EC. Concentrations of total proteins, Z/ZR (Zeatin/zeatin riboside) and iP/iPA (N6-(Δ2-isopentenyl) adenine and N6-(Δ2-isopentenyl) adenosine) were similar in EC and NEC.

La explotación intensiva de la madera de caoba (Swietenia macrophylla) ha provocado la pérdida de poblaciones naturales. La embriogénesis somática ofrece una alternativa a la propagación clonal y la conservación de esta especie. Este estudio describe las características bioquímicas (contenido de carbohidratos, fenoles totales, flavonoides totales, proteínas y reguladores del crecimiento) e histológicas del proceso de embriogénesis somática en caoba. Los callos se obtuvieron cultivando cotiledones de semillas de frutos inmaduros durante seis semanas en medio MS semisólido suplementado con 1.0 mgL-1 de kinetina y 4.0 mgL-1 de 2, 4-D. Luego se cultivó el callo primario en medio MS semisólido y un suplemento de 1.0 mgl-1 BA y se obtuvieron estructuras embriogénicas. El desarrollo de embriones somáticos de forma globular a la etapa cotiledonar se confirmó por histología y microscopía electrónica de barrido. La iniciación del brote se observó después de que los embriones somáticos se transfirieron a un medio de germinación y maduración. Se determinaron las concentraciones endógenas de carbohidratos, fenoles totales, flavonoides totales, proteínas y reguladores del crecimiento en callos embriogénicos (EC) y no embriogénicos (NEC) de caoba. Los cultivos embriogénicos contenían concentraciones significativamente más altas de IAA, ABA y GA, mientras que los callos no embriogénicos contenían más fenoles totales, flavonoides y almidón resistente. La fructosa y la glucosa no estaban presentes en niveles detectables en EC o NEC, mientras que el almidón soluble y la sacarosa solo se detectaron en el EC. Las concentraciones de proteínas totales, Z / ZR e iP / iPA fueron similares en EC y NEC.

Single nucleotide polymorphism analysis reveals heterogeneity within a seedling tree population of a polyembryonic mango cultivar.

Within a polyembryonic mango seedling tree population, the genetic background of individuals should be identical because vigorous plants for cultivation are expected to develop from nucellar embryos representing maternal clones. Due to the fact that the mango cultivar 'Hôi' is assigned to the polyembryonic ecotype, an intra-cultivar variability of ethylene receptor genes was unexpected. Ethylene receptors in plants are conserved, but the number of receptors or receptor isoforms is variable regarding different plant species. However, it is shown here that the ethylene receptor MiETR1 is present in various isoforms within the mango cultivar 'Hôi'. The investigation of single nucleotide polymorphisms revealed that different MiETR1 isoforms can not be discriminated simply by individual single nucleotide exchanges but by the specific arrangement of single nucleotide polymorphisms at certain positions in the exons of MiETR1. Furthermore, an MiETR1 isoform devoid of introns in the genomic sequence was identified. The investigation demonstrates some limitations of high resolution melting and ScreenClust analysis and points out the necessity of sequencing to identify individual isoforms and to determine the variability within the tree population.