Analysis of the peel structure of different Citrus spp. via light microscopy, SEM and µCT with manual and automatic segmentation.

The peels of lime, lemon, pomelo and citron are investigated at macroscopic and microscopic level. The structural composition of the peels is compared and properties such as peel thickness, proportion of flavedo, density and proportion of intercellular spaces are determined. µCT images are used to visualize vascular bundles and oil glands. SEM images provide information about the appearance of the cellular tissue in the outer flavedo and inner albedo. The proportion of intercellular spaces is quantitatively determined by manual and software-assisted analysis (ilastik). While there are macroscopic differences in the fruits, they differ only slightly in the orientation of the vascular bundles and the arrangement of the oil glands. However, in peel thickness and flavedo thickness, the fruit peels differ significantly from each other. There are no significant differences between the two analysis methods used, although the use of ilastik is preferred due to time reduction of up to 70%. The large amount of intercellular spaces in the albedo but also the denser flavedo both have a mechanical protective function to prevent damage to the fruit. In addition, the entire peel structure is mechanically reinforced by vascular bundles. This combination of penetration protection (flavedo) and energy dissipation (albedo) makes Citrus spp. peels a promising inspiration for technical material systems.

New roles of NO TRANSMITTING TRACT and SEEDSTICK during medial domain development in Arabidopsis fruits.

The gynoecium, the female reproductive part of the flower, is key for plant sexual reproduction. During its development, inner tissues such as the septum and the transmitting tract tissue, important for pollen germination and guidance, are formed. In Arabidopsis, several transcription factors are known to be involved in the development of these tissues. One of them is NO TRANSMITTING TRACT (NTT), essential for transmitting tract formation. We found that the NTT protein can interact with several gynoecium-related transcription factors, including several MADS-box proteins, such as SEEDSTICK (STK), known to specify ovule identity. Evidence suggests that NTT and STK control enzyme and transporter-encoding genes involved in cell wall polysaccharide and lipid distribution in gynoecial medial domain cells. The results indicate that the simultaneous loss of NTT and STK activity affects polysaccharide and lipid deposition and septum fusion, and delays entry of septum cells to their normal degradation program. Furthermore, we identified KAWAK, a direct target of NTT and STK, which is required for the correct formation of fruits in Arabidopsis These findings position NTT and STK as important factors in determining reproductive competence.

Transcriptome and Metabolome Analyses Provide Insights into the Watercore Disorder on "Akibae" Pear Fruit.

Watercore is a physiological disorder that commonly occurs in sand pear cultivars. The typical symptom of watercore tissue is transparency, and it is often accompanied by browning, breakdown and a bitter taste during fruit ripening. To better understand the molecular mechanisms of watercore affecting fruit quality, this study performed transcriptome and metabolome analyses on watercore pulp from "Akibae" fruit 125 days after flowering. The present study found that the "Akibae" pear watercore pulp contained higher sorbitol and sucrose than healthy fruit. Moreover, the structure of the cell wall was destroyed, and the content of pectin, cellulose and hemicellulose was significantly decreased. In addition, the content of ethanol and acetaldehyde was significantly increased, and the content of polyphenol was significantly decreased. Watercore induced up-regulated expression levels of sorbitol synthesis-related (sorbitol-6-phosphate dehydrogenase, S6PDH) and sucrose synthesis-related genes (sucrose synthesis, SS), whereas it inhibited the expression of sorbitol decomposition-related genes (sorbitol dehydrogenase, SDH) and sorbitol transport genes (sorbitol transporter, SOT). Watercore also strongly induced increased expression levels of cell wall-degrading enzymes (polygalactosidase, PG; ellulase, CX; pectin methylesterase, PME), as well as ethanol synthesis-related (alcohol dehydrogenase, ADH), acetaldehyde synthesis-related (pyruvate decarboxylase, PDC) and polyphenol decomposition-related genes (polyphenol oxidase, PPO). Moreover, the genes that are involved in ethylene (1-aminocyclopropane- 1-carboxylate oxidase, ACO; 1-aminocyclopropane- 1-carboxylate synthase, ACS) and abscisic acid (short-chain alcohol dehydrogenase, SDR; aldehyde oxidase, AAO) synthesis were significantly up-regulated. In addition, the bitter tasting amino acids, alkaloids and polyphenols were significantly increased in watercore tissue. Above all, these findings suggested that the metabolic disorder of sorbitol and sucrose can lead to an increase in plant hormones (abscisic acid and ethylene) and anaerobic respiration, resulting in aggravated fruit rot and the formation of bitter substances.

Carotenogenesis and chromoplast development during ripening of yellow, orange and red colored Physalis fruit.

MAIN CONCLUSION: Formation of specific ultrastructural chromoplastidal elements during ripening of fruits of three different colored Physalis spp. is closely related to their distinct carotenoid profiles. The accumulation of color-determining carotenoids within the chromoplasts of ripening yellow, orange, and red fruit of Physalis pubescens L., Physalis peruviana L., and Physalis alkekengi L., respectively, was monitored by high-performance liquid chromatography/diode array detector/tandem mass spectrometry (HPLC-DAD-MS/MS) as well as light and transmission electron microscopy. Both yellow and orange fruit gradually accumulated mainly ß-carotene and lutein esters at variable levels, explaining their different colors at full ripeness. Upon commencing ß-carotene biosynthesis, large crystals appeared in their chromoplasts, while large filaments protruding from plastoglobules were characteristic elements of chromoplasts of orange fruit. In contrast to yellow and orange fruit, fully ripe red fruit contained almost no ß-carotene, but esters of both ß-cryptoxanthin and zeaxanthin at very high levels. Tubule bundles and unusual disc-like crystallites were predominant carotenoid-bearing elements in red fruit. Our study supports the earlier hypothesis that the predominant carotenoid type might shape the ultrastructural carotenoid deposition form, which is considered important for color, stability and bioavailability of the contained carotenoids.

Transcription factor CaNAC1 regulates low-temperature-induced phospholipid degradation in green bell pepper.

Phospholipids constitute the main component of biomembranes. During low-temperature storage and transportation of harvested bell peppers (Capsicum annuum), chilling injury participates in their decay. A primary cause of this chilling injury is phospholipid degradation. In this study, three genes encoding phospholipase D (PLD) were identified from bell peppers and their activities were examined under cold stress. Low temperature (4 °C) induced strong accumulation of the CaPLDα4 transcript, suggesting that it is associated with the phenomenon of phospholipid degradation and destruction of cell membranes. Low temperature also significantly induced increased amounts of NAM-ATAF1/2-CUC2 (NAC) domain transcription factors. CaNAC1 was found to interact with the promoter of CaPLD4 in a yeast one-hybrid screen. Electrophoretic mobility shift and ß-glucuronidase reporter assays demonstrated that CaNAC1 binds to the CTGCAG motif in the CaPLDα4 promoter, thereby activating its transcription and controlling phospholipid degradation. The ubiquitination sites of the CaNAC1 protein were characterized by liquid chromatography-tandem mass spectrometry. We conclude that CaNAC1 is a transcriptional activator of CaPLDα4 and suggested that it participates in the degradation of membrane lipids in bell peppers when they are stored at low temperature.

Anatomical Investigation and GC/MS Analysis of 'Coco de Mer', Lodoicea maldivica (Arecaceae).

Lodoicea maldivica (J.F.Gmel.) Pers. (Arecaceae), 'Coco de Mer', is a palm, growing as endemic in the Seychelles islands. Its fruit weighs up to 20 kg and is characterized by a fleshy and fibrous envelope surrounding the nutlike portion. The present work combines a morpho-anatomical and a phytochemical analysis of the fruit exocarp and mesocarp. The exocarp is composed by a layer of palisade cells. The mesocarp is characterized by vascular bundles and by sclereids. In the aerenchyma, the internal zone of the mesocarp, cells aggregates were positive to phenols, while idioblasts were positive to terpenes. We performed a GC/MS analysis with a semi-quantitative relative amount calculation of the recorded compounds. The GC/MS essential oil profile revealed the dominance of acyclic sesquiterpenoids (53.95 %), followed by bicyclic sesquiterpenoids (31.69 %), monoterpenes (11.89 %) and monocyclic sesquiterpenoids (2.44 %). The terpenes detected in higher amounts, ß-caryophyllene and bicyclogermacrene, are known for activity against insect larvae, but have been proposed as antiviral candidates against SARS-CoV-2. The third compound in amount, aromadendrene, is active against bacteria and, again, known to possess insecticidal properties.

Physicochemical changes and chilling injury disorders in 'Tango' mandarins stored at low temperatures.

BACKGROUND: The susceptibility to chilling injury and quality changes of 'Tango' mandarins stored at different temperatures was evaluated in fruit grown at two locations in Andalusia (Spain) and grafted on Carrizo Citrange or FA5 rootstock. The peel disorders were also characterized by a microstructural study. RESULTS: Fruit developed chilling injuries, manifested as pitting lesions affecting the equatorial area of the fruit stored at 1 °C or 5 °C; fruit growing on FA5 rootstock showed a slightly lower incidence. The microstructural study revealed that only the upper layers of flavedo were affected in the damaged fruit, the epidermal and hypodermal tissues being dramatically collapsed. Although the fruit was prone to accumulate ethanol, especially after the shelf life that followed the different periods of cold storage, the ethanol did not compromise the overall flavor. CONCLUSIONS: Storage of 'Tango' fruit was limited by chilling injuries when stored at 1 °C or 5 °C for more than 20 days. Moreover, at these temperatures, the fruit was prone to accumulate ethanol and develop off flavors. At 9° C, the fruit could be stored for 30 days without compromising external or internal quality. Growing location and rootstock influenced some quality attributes at harvest but not during storage. © 2020 Society of Chemical Industry.

Transcriptomic and functional analysis of cucumber (Cucumis sativus L.) fruit phloem during early development.

The phloem of the Cucurbitaceae has long been a subject of interest due to its complex nature and the economic importance of the family. As in a limited number of other families, cucurbit phloem is bicollateral, i.e. with sieve tubes on both sides of the xylem. To date little is known about the specialized functions of the internal phloem (IP) and external phloem (EP). Here, a combination of microscopy, fluorescent dye transport analysis, micro-computed tomography, laser capture microdissection and RNA-sequencing (RNA-Seq) were used to study the functions of IP and EP in the vascular bundles (VBs) of cucumber fruit. There is one type of VB in the peduncle, but four in the fruit: peripheral (PeVB), main (MVB), carpel (CVB) and placental (PlVB). The VBs are bicollateral, except for the CVB and PlVB. Phloem mobile tracers and 14 C applied to leaves are transported primarily in the EP, and to a lesser extent in the IP. RNA-Seq data indicate preferential gene transcription in the IP related to differentiation/development, hormone transport, RNA or protein modification/processing/transport, and nitrogen compound metabolism and transport. The EP preferentially expresses genes for stimulus/stress, defense, ion transport and secondary metabolite biosynthesis. The MVB phloem is preferentially involved in photoassimilate transport, unloading and long-distance signaling, while the PeVB plays a more substantial role in morphogenesis and/or development and defense response. CVB and PlVB transcripts are biased toward development of reproductive organs. These findings provide an integrated view of the differentiated structure and function of the vascular tissue in cucumber fruit.

Russeting partially restores apple skin permeability to water vapour.

MAIN CONCLUSION: The higher water loss of russeted fruit results from the higher permeance of the periderm of the russeted skin as compared to that of the intact cuticle and epidermis. Apple fruit surfaces are often in-parallel composites, comprising areas of intact cuticle (atop a healthy epidermis) adjacent to areas covered by periderm (so-called russet). The occurrence of non-russeting and russeting genotypes makes this species an ideal model to study the barrier properties of its composite skin. The objective was to quantify the water vapour permeances of non-russeted ([Formula: see text]) and russeted fruit skins ([Formula: see text]). Rates of water loss from whole fruit ([Formula: see text]) and excised epidermal skin segments (ES) or peridermal skin segments (PS) were quantified gravimetrically. The [Formula: see text] was larger in russeting than in non-russeting genotypes because [Formula: see text] exceeded [Formula: see text] by about twofold. Also, the [Formula: see text] of russeting genotypes was larger than that of non-russeting genotypes. Generally, [Formula: see text] was more variable than [Formula: see text]. These differences were consistent across seasons and genotypes. The lower [Formula: see text] as compared to [Formula: see text] resulted primarily from the higher wax content of the cuticle of the [Formula: see text]. For non-russeted genotypes, the value of [Formula: see text] was significantly related to the permeance determined on the same intact fruit ([Formula: see text]). Close relationships were also found between the [Formula: see text] calculated from [Formula: see text] determined on the same fruit and the measured [Formula: see text]. For russeting genotypes, the [Formula: see text] or [Formula: see text] were not correlated with [Formula: see text]. The [Formula: see text] calculated from [Formula: see text], [Formula: see text], [Formula: see text] and [Formula: see text] (all determined on an individual-fruit basis) was significantly correlated with the measured [Formula: see text]. Our results demonstrate that the periderm permeance exceeds the cuticle permeance and that permeances of non-russeted surfaces of russeting genotypes exceed those of non-russeting genotypes.

Classification of fruit trichomes in cucumber and effects of plant hormones on type II fruit trichome development.

MAIN CONCLUSION: Cucumber fruit trichomes could be classified into eight types; all of them are multicellular with complex and different developmental processes as compared with unicellular trichomes in other plants. The fruit trichomes or fruit spines of cucumber, Cucumis sativus L., are highly specialized structures originating from epidermal cells with diverse morphology, which grow perpendicular to the fruit surface. To understand the underlying molecular mechanisms of fruit trichome development, in this study, we conducted morphological characterization and classification of cucumber fruit trichomes and their developmental processes. We examined the fruit trichomes among 200 cucumber varieties, which could be classified into eight morphologically distinct types (I-VIII). Investigation of the organogenesis of the eight types of trichomes revealed two main developmental patterns. The development of glandular trichomes had multiple stages including initiation and expansion of the trichome precursor cell protuberating out of the epidermal surface, followed by periclinal bipartition to two cells (top and bottom) which later formed the head region and the stalk, respectively, through subsequent cell divisions. The non-glandular trichome development started with the expansion of the precursor cell perpendicularly to the epidermal plane followed by cell periclinal division to form a stalk comprising of some rectangle cells and a pointed apex cell. The base cell then started anticlinal bipartition to two cells, which then underwent many cell divisions to form a multicellular spherical structure. In addition, phytohormones as environmental cues were closely related to trichome development. We found that GA and BAP were capable of increasing trichome number per fruit with distinct effects under different concentrations.

Origin of the Taxaceae aril: evolutionary implications of seed-cone teratologies in Pseudotaxus chienii.

Background and Aims: Fleshy structures that promote biotic dispersal by ingestion have evolved many times in seed plants. Within the yew family Taxaceae sensu lato (six genera, including Cephalotaxus), it remains controversial whether the characteristic fleshy structure surrounding the seed is interpreted as a novel outgrowth of the base of the ovule (i.e. an aril) or a fleshy seed coat that is entirely derived from the integument (i.e. a sarcotesta). This paper presents a detailed study of both wild-type and teratological seed cones of Pseudotaxus chienii, including morphology, anatomy and ontogeny. Methods: Wild-type and teratological seed cones were investigated with the classical paraffin technique and subsequent astrablue/safranin staining and scanning electron microscopy. Key Results: The wild-type seed cone of Pseudotaxus possesses a fleshy white aril that is cup-like and not entirely fused to the seed. In the teratological seed cones investigated, the aril was bilobed and consisted of two free halves. In both wild-type and teratological cones, the aril was initiated as two lateral primordia in a transverse plane, but in wild-type cones the two primordia became extended into a ring primordium, which grew apically, leading to the cup-like shape. The teratological seed cones lacked a ring primordium and the two lateral aril lobes remained free throughout their entire ontogeny, alternating with the scale-like leaves inserted below them on the same branch; in some cases, these leaves also became fleshy. Conclusions: Based on the ontogeny and arrangement of the two fleshy aril lobes in the teratological seed cones of Pseudotaxus, we suggest that the typical aril of Taxaceae could be readily interpreted as a fused pair of strongly swollen leaves rather than a modified integument. Our investigations of the cup-like aril of Pseudotaxus demonstrate a similarity not only with other Taxaceae but also with relatively distantly related conifers such as Phyllocladus (Podocarpaceae).

Cretaceous asterid evolution: fruits of Eydeia jerseyensis sp. nov. (Cornales) from the upper Turonian of eastern North America.

BACKGROUND AND AIMS: The asterids (>80 000 extant species) appear in the fossil record with considerable diversity near the Turonian-Coniacian boundary (~90 Ma; Late Cretaceous) and are strongly represented in the earliest diverging lineage, Cornales. These early asterid representatives have so far been reported from western North America and eastern Asia. In this study, we characterize a new cornalean taxon based on charcoalified fruits from the upper Turonian of eastern North America, a separate landmass from western North America at the time, and identify early palaeobiogeographical patterns of Cornales during the Cretaceous. METHODS: Fossils were studied and imaged using scanning electron microscopy and micro-computed tomography (micro-CT) scanning. To assess the systematic affinities of the fossils, phylogenetic analyses were conducted using maximum parsimony. KEY RESULTS: The charcoalified fruits are represented by tri-locular woody endocarps with dorsal apically opening germination valves. Three septa intersect to form a robust central axis. Endocarp ground tissue consists of two zones: an outer endocarp composed of isodiametric sclereids and an inner endocarp containing circum-locular fibres. Central vasculature is absent; however, there are several small vascular bundles scattered within the septa. Phylogenetic analysis places the new taxon within the extinct genus Eydeia. DISCUSSION: Thick-walled endocarps with apically opening germination valves, no central vascular bundle and one seed per locule are indicative of the order Cornales. Comparative analysis suggests that the fossils represent a new species, Eydeia jerseyensis sp. nov. This new taxon is the first evidence of Cornales in eastern North America during the Cretaceous and provides insights into the palaeobiogeography and initial diversification of the order.

Methyl salicylate delays peel yellowing of 'Zaosu' pear (Pyrus bretschneideri) during storage by regulating chlorophyll metabolism and maintaining chloroplast ultrastructure.

BACKGROUND: In some cultivars, yellowing resulting from chlorophyll breakdown has a direct and negative effect on food supply and health. The 'Zaosu' pear (Pyrus bretschneideri Rehd.), a commercial Asian pear cultivar in China, rapidly turns yellow when stored at room temperature after harvest. To develop techniques that delay or suppress chlorophyll degradation, the effects of methyl salicylate (MeSA) on yellowing in 'Zaosu' pear fruit during storage were evaluated. RESULTS: Compared with the untreated fruit, the application of 0.05 mmol L-1 MeSA delayed the decline of the total chlorophyll, chlorophyll a and chlorophyll b content, and maintained more intact chloroplasts with fewer and smaller plastoglobuli. Methyl salicylate suppressed enzyme activities, including chlorophyllase, chlorophyll-degrading peroxidase, Mg dechelatase, and pheophytinase, and the expression levels of NYC, NOL, CLH, SGR, PPH, PAO and RCCR in treated fruit. CONCLUSION: Methyl salicylate could delay chlorophyll breakdown in the fruit. The results also suggested that the conversion from chlorophyll a to pheophorbide a could proceed via two pathways, and that alternative pathways for the breakdown of chlorophyll a exist in 'Zaosu' pears. © 2019 Society of Chemical Industry.

Vesicles Are Persistent Features of Different Plastids.

Peripheral vesicles in plastids have been observed repeatedly, primarily in proplastids and developing chloroplasts, in which they are suggested to function in thylakoid biogenesis. Previous observations of vesicles in mature chloroplasts have mainly concerned low temperature pretreated plants occasionally treated with inhibitors blocking vesicle fusion. Here, we show that such vesicle-like structures occur not only in chloroplasts and proplastids, but also in etioplasts, etio-chloroplasts, leucoplasts, chromoplasts and even transforming desiccoplasts without any specific pretreatment. Observations are made both in C3 and C4 species, in different cell types (meristematic, epidermis, mesophyll, bundle sheath and secretory cells) and different organs (roots, stems, leaves, floral parts and fruits). Until recently not much focus has been given to the idea that vesicle transport in chloroplasts could be mediated by proteins, but recent data suggest that the vesicle system of chloroplasts has similarities with the cytosolic coat protein complex II system. All current data taken together support the idea of an ongoing, active and protein-mediated vesicle transport not only in chloroplasts but also in other plastids, obviously occurring regardless of chemical modifications, temperature and plastid developmental stage.

Patterns of microcracking in apple fruit skin reflect those of the cuticular ridges and of the epidermal cell walls.

MAIN CONCLUSION: Microcracks in the cuticle of developing apples are aligned with ridges on the inner cuticle surface and are indicative of stress-strain concentrations above the anticlinal cell walls. Microcracks occur in cuticles of most fruits. Growth strains are considered causal. In apples (Malus × domestica), microcracks usually form a mesh pattern similar to that formed by cuticular ridges. Ridge patterns are similar to those of the epidermal cells' anticlinal walls. Our aim was to identify the mechanistic bases for these pattern similarities. By quantifying ridge depth, ridge width, and the areas enclosed by ridges, we reveal the presence of major and minor ridges. Major ridges enclose two-to-four epidermal cells, minor ridges only one cell. There are similar and overlying patterns of microcracking on the cuticle's outer surface and of ridges on its inner surface-microcracks generally follow the outlines of the major ridges. In biaxial tensile tests at 20 kPa, strains were low and microcracks shallow, but at > 40 kPa, strains were higher and microcracks deeper. Microcracks traversing the cuticle are usually aligned with the anticlinal walls of the underlying epidermal cells. In general, increased skin strain is associated with increased skin transpiration. Transpiration increases are reversible for low strains but irreversible for high strains. The alignment of cuticular microcracks with the major ridges, and these with the anticlinal cell walls, indicates associated stress/strain concentrations.

Cuticle Biosynthesis in Tomato Leaves Is Developmentally Regulated by Abscisic Acid.

The expansion of aerial organs in plants is coupled with the synthesis and deposition of a hydrophobic cuticle, composed of cutin and waxes, which is critically important in limiting water loss. While the abiotic stress-related hormone abscisic acid (ABA) is known to up-regulate wax accumulation in response to drought, the hormonal regulation of cuticle biosynthesis during organ ontogeny is poorly understood. To address the hypothesis that ABA also mediates cuticle formation during organ development, we assessed the effect of ABA deficiency on cuticle formation in three ABA biosynthesis-impaired tomato mutants. The mutant leaf cuticles were thinner, had structural abnormalities, and had a substantial reduction in levels of cutin. ABA deficiency also consistently resulted in differences in the composition of leaf cutin and cuticular waxes. Exogenous application of ABA partially rescued these phenotypes, confirming that they were a consequence of reduced ABA levels. The ABA mutants also showed reduced expression of genes involved in cutin or wax formation. This difference was again countered by exogenous ABA, further indicating regulation of cuticle biosynthesis by ABA. The fruit cuticles were affected differently by the ABA-associated mutations, but in general were thicker. However, no structural abnormalities were observed, and the cutin and wax compositions were less affected than in leaf cuticles, suggesting that ABA action influences cuticle formation in an organ-dependent manner. These results suggest dual roles for ABA in regulating leaf cuticle formation: one that is fundamentally associated with leaf expansion, independent of abiotic stress, and another that is drought induced.

Fossil moonseeds from the Paleogene of West Gondwana (Patagonia, Argentina).

PREMISE OF THE STUDY: The fossil record is critical for testing biogeographic hypotheses. Menispermaceae (moonseeds) are a widespread family with a rich fossil record and alternative hypotheses related to their origin and diversification. The family is well-represented in Cenozoic deposits of the northern hemisphere, but the record in the southern hemisphere is sparse. Filling in the southern record of moonseeds will improve our ability to evaluate alternative biogeographic hypotheses. METHODS: Fossils were collected from the Salamanca (early Paleocene, Danian) and the Huitrera (early Eocene, Ypresian) formations in Chubut Province, Argentina. We photographed them using light microscopy, epifluorescence, and scanning electron microscopy and compared the fossils with similar extant and fossil Menispermaceae using herbarium specimens and published literature. KEY RESULTS: We describe fossil leaves and endocarps attributed to Menispermaceae from Argentinean Patagonia. The leaves are identified to the family, and the endocarps are further identified to the tribe Cissampelideae. The Salamancan endocarp is assigned to the extant genus Stephania. These fossils significantly expand the known range of Menispermaceae in South America, and they include the oldest (ca. 64 Ma) unequivocal evidence of the family worldwide. CONCLUSIONS: Our findings highlight the importance of West Gondwana in the evolution of Menispermaceae during the Paleogene. Currently, the fossil record does not discern between a Laurasian or Gondwanan origin; however, it does demonstrate that Menispermaceae grew well outside the tropics by the early Paleocene. The endocarps' affinity with Cissampelideae suggests that diversification of the family was well underway by the earliest Paleocene.

Antagonistic Activity and the Mechanism of Bacillus amyloliquefaciens DH-4 Against Citrus Green Mold.

Citrus fruit usually suffer significant losses during the storage and transportation stages. Green mold, a postharvest rot of citrus fruit caused by Penicillium digitatum, is one of the most serious fungal diseases. In this study, the antagonist strain DH-4 was identified as Bacillus amyloliquefaciens according to morphological observation and 16S ribosomal DNA analysis. In addition, it showed broad antifungal activity, especially the suppression of Penicillium spp. The culture filtrate of strain DH-4 exhibited apparent activity against P. digitatum in vitro and in vivo. In storage, the culture filtrate with DH-4 in it showed a better antiseptic effect. The antifungal substances in the culture filtrate, produced by strain DH-4, displayed stable activity in various extreme conditions. In addition, the antifungal substances in the culture filtrate were identified as macrolactin, bacillaene, iturins, fengycin, and surfactin by ultraperformance liquid chromatography (UPLC) electrospray ionization mass spectrometry analysis. The UPLC fractions containing these antifungal compounds were basically heat tolerant and all responsible for the antagonistic activity against P. digitatum. Transmission electron microscope observation indicated that the antifungal substances might cause abnormalities in the P. digitatum cellular ultrastructure, which could be the possible mode of action of B. amyloliquefaciens against P. digitatum. In addition, it was confirmed via scanning electron microscope analysis that the main way it inhibited P. digitatum was by secreting antimicrobial compounds without direct interaction. This study contributes to the understanding of the mechanism of B. amyloliquefaciens against citrus green mold as well as providing a potential application for the biocontrol of postharvest rot diseases in citrus fruit.

Physiological, molecular and ultrastructural analyses during ripening and over-ripening of banana (Musa spp., AAA group, Cavendish sub-group) fruit suggest characteristics of programmed cell death.

BACKGROUND: Programmed cell death (PCD) is a part of plant development that has been studied for petal senescence and vegetative tissue but has not been thoroughly investigated for fleshy fruits. The purpose of this research was to examine ripening and over-ripening in banana fruit to determine if there were processes in common to previously described PCD. RESULTS: Loss of cellular integrity (over 40%) and development of senescence related dark spot (SRDS) occurred after day 8 in banana peel. Nuclease and protease activity in the peel increased during ripening starting from day 2, and decreased during over-ripening. The highest activity was for proteases and nucleases with apparent molecular weights of 86 kDa and 27 kDa, respectively. Images of SRDS showed shrinkage of the upper layers of cells, visually suggesting cell death. Decrease of electron dense areas was evident in TEM micrographs of nuclei. CONCLUSION: This study shows for the first time that ripening and over-ripening of banana peel share physiological and molecular processes previously described in plant PCD. SRDS could represent a morphotype of PCD that characterizes a structural and biochemical failure in the upper layers of the peel, thereafter spreading to lower and adjacent layers of cells. © 2017 Society of Chemical Industry.

Tannosomes in the Pericarp Cells of Maloideae (Rosaceae).

Data on the presence of structural units termed tannosomes in the pericarp cells of Maloideae has been obtained for the first time. Tannosomes merge in the vacuoles to form "tannoglobules."