Structure and functioning of oil cavities in the shoot apex of Metrodorea nigra A. St.-Hil. (Rutaceae).

This study investigates the histology and subcellular features of secretory cavities during the development of the shoot apex of Metrodorea nigra A. St.-Hil. in order to better understand the functioning of these glands. This Rutaceae species is a very suitable model for studying secretory cavity life span, since the shoot apex exhibits both dormant and growth stages during its annual cycle. Shoot apices were collected during the dormant and growth stages from populations of M. nigra growing under natural conditions. Materials were processed using standard techniques for light and electron microscopy. The secretory cavities originate under the protodermis, and their initiation is restricted to the early developmental stage of shoot organs, which are protected by a hood-shaped structure. Secretory cavities have a multi-seriate epithelium surrounding a lumen that expands schizolysigenously. Oil production begins before lumen formation. When the shoot apex resumes development after the dormant stage, the glands remain active in oil secretion in the developing shoot apex and fully expanded leaves. The mature epithelial cells are flattened and exhibit very thin walls, large oil bodies, leucoplasts surrounded by endoplasmic reticulum, and mitochondria with unusual morphology. The tangential walls of the epithelial cells facing the lumen undergo continuous peeling. The vacuole extrusion appears to be the primary mode of release oil into the lumen, in an exocytotic way. The continuity of oil secretion is ensured by the replacement of the damaged inner epithelial cells by divisions in the parenchyma layer that surround the oil gland, likely a meristematic sheath.

Carotenoid accumulation affects redox status, starch metabolism, and flavonoid/anthocyanin accumulation in citrus.

BACKGROUND: Carotenoids are indispensable plant secondary metabolites that are involved in photosynthesis, antioxidation, and phytohormone biosynthesis. Carotenoids are likely involved in other biological functions that have yet to be discovered. In this study, we integrated genomic, biochemical, and cellular studies to gain deep insight into carotenoid-related biological processes in citrus calli overexpressing CrtB (phytoene synthase from Pantoea agglomerans). Fortunella hindsii Swingle (a citrus relative) and Malus hupehensis (a wild apple) calli were also utilized as supporting systems to investigate the effect of altered carotenoid accumulation on carotenoid-related biological processes. RESULTS: Transcriptomic analysis provided deep insight into the carotenoid-related biological processes of redox status, starch metabolism, and flavonoid/anthocyanin accumulation. By applying biochemical and cytological analyses, we determined that the altered redox status was associated with variations in O2 (-) and H2O2 levels. We also ascertained a decline in starch accumulation in carotenoid-rich calli. Furthermore, via an extensive cellular investigation of the newly constructed CrtB overexpressing Fortunella hindsii Swingle, we demonstrated that starch level reducation occurred in parallel with significant carotenoid accumulation. Moreover, studying anthocyanin-rich Malus hupehensis calli showed a negative effect of carotenoids on anthocyanin accumulation. CONCLUSIONS: In citrus, altered carotenoid accumulation resulted in dramatic effects on metabolic processes involved in redox modification, starch degradation, and flavonoid/anthocyanin biosynthesis. These findings provided new perspectives to understand the biological importance of carotenogenesis and of the developmental processes associated with the nutritional and sensory qualities of agricultural products that accumulate carotenoids.

ULTRASTRUCTURAL RESPONSE OF EMBRYONIC AXES OF Fortunella polyandra TO DEHYDRATION AND CRYOPRESERVATION.

BACKGROUND: To further understand the survival characteristics of desiccation-sensitive excised embryonic axes of Fortunella polyandra to desiccation and cryopreservation it is necessary to study the impact of drying rates on both the ultrastructure and electrolyte leakage. OBJECTIVE: To examine the effects of two different drying regimes (silica gel and ultra-rapid) on the survival, ultrastructure and membrane leakage characteristics of excised embryonic axes of F. polyandra before and after cryopreservation. MATERIALS AND METHODS: The effects of the drying regimes on the survival, ultrastructure and membrane integrity of the excised embryonic axes of F. polyandra was determined. Survival was assessed in vitro, and the integrity of membranes following drying was estimated by electrolyte leakage and observation under the transmission electron microscope (TEM). Survival and ultrastructural changes were also observed after cryopreservation. RESULTS: Electrolyte leakage increased with decreasing water content of the embryonic axes, indicative of substantial subcellular damage, after both ultra-rapid dehydration (to water contents of <0.16 g H2O g(-1) dw) and silica gel dehydration (to <0.28 g H2O g(-1) dw water content). Ultrastructurally, axes showed increasing cytoplasm and vacuole shrinkage and disruption of cell membranes with longer dehydration periods. Normal seedling recovery of 50 to 47% for cryopreserved embryonic axes of F. polyandra was observed after ultra-rapid and silica gel drying respectively. Extreme cell injury was observed after exposure to liquid nitrogen at high moisture content. Although cells of dehydrated axes encountered stress during cryopreservation, the main damage occurred during the dehydration step. CONCLUSION: For surviving axes, the damage was less severe and the axes grew to become normal seedlings. Ultrastructural studies reveal the damage of the cells at different rates of dehydration and during cryopreservation.

Fusion within and between whorls of floral organs in Galipeinae (Rutaceae): structural features and evolutionary implications.

BACKGROUND AND AIMS: Most genera of the neotropical Galipeinae (tribe Galipeeae, Rutoideae) exhibit several forms and degrees of fusion between the floral organs, including the union of petals into an apparently sympetalous corolla, the joining of the stamens among themselves and to the corolla, and the partial to complete connation of carpels. Though these and others floral traits are currently used in the circumscription of species in Galipeinae, few studies have shown in detail in which way (postgenital or congenital) and to what extent these fusions occur. To elucidate these anatomical conditions, a structural study of the flowers of the Galipeinae species was carried out. METHODS: Flowers of six species from three genera of Galipeinae were studied in their morphology, anatomy and development with stereomicroscopy, light microscopy and scanning electron microscopy (SEM). KEY RESULTS: The floral tube is formed by synorganization of stamens with petals in all species, and exhibits three main patterns: (1) Conchocarpus heterophyllus and C. minutiflorus have a floral tube formed by marginal coherence/adherence of petals and filaments due to interwining trichomes (postgenital connection); (2) Erythrochiton brasiliensis has a tube formed by congenital fusion of petals and filaments; and (3) Galipea jasminiflora and Conchocarpus macrophyllus have a tube formed distally with the first pattern, and proximally with the second pattern. Although floral tubes seem to be homologous within Galipeinae, this is not true at the level of the family: the floral tube of Correa (from an only distantly related clade of the family) is formed by postgenital union of the petals representing a convergent structure. The gynoecium of the studied species of Galipeinae shows a great variability in the extent of fusion of carpel flanks. Even though different structures for the mature gynoecium were found in each genus, all genera show postgenitally fused carpel apices, which is related to the formation of a compitum, as described earlier for other members of Rutaceae. CONCLUSIONS: The degree and diversity of fusions of floral organs in Galipeinae is unique within the order Sapindales. A study of the amount of diversification of Galipeinae in South America and comparison with other clades of Rutaceae would be of interest.

Floral ontogeny of Ruteae (Rutaceae) and its systematic implications.

Floral development was investigated in Ruta graveolens and Psilopeganum sinense, representing two genera in the tribe Ruteae. Special attention was paid to the sequence of initiation of organ whorls in the androecium and gynoecium. The antepetalous stamens arise at the same level as the antesepalous stamens in both species. The carpels are antepetalous in both taxa, indicating the androecium in both genera is obdiplostemonous. Compared with floral ontogeny of the ancestral genus Phellodendron (Toddalioideae), the obdiplostemonous androecium is a derived condition. The floral apex in P. sinense is quadrangular before initiation of the two carpels. Additionally, there are four dorsal and four ventral traces in the ovary. Integrated morphological and anatomical evidence indicates that the bicarpellate gynoecium in Psilopeganum most likely evolved from a tetracarpellate ancestor. Considering the similarities in morphological, geographical and chromosomal features, the ancestor may be Ruta-like. Further molecular phylogenetic and genetic studies are needed to verify this assumption.

Pollen morphology of the genus Skimmia (Rutaceae) and its taxonomic implications.

Pollen morphology of the genus Skimmia was studied. Of six species of the genus, five species were investigated by light microscopy and scanning electron microscopy. Pollen grains of Skimmia had striate to striate-reticulate sculpture and variable aperture number (3-7), and these characters differed from even the phylogenetically closest relatives such as Dictamnus and Casimiroa which have 3-colporate pollen grains with reticulate exine. In Skimmia laureola, pollen dimorphism is suggested in the aperture number in accordance with the floral system. Pollen grains of Skimmia japonica, a morphologically very variable species, were extensively surveyed covering its whole distribution area. The grains were also striate to striate-reticulate, and the exine sculpture was not different between varieties. However, the aperture number of S. japonica showed a geographical variation ranging from 4 to 6; in Sakhalin, Hokkaido, most part of Honshu and Taiwan, grains with 4-5 apertures were dominant, while in the southern part of Japan, including the Ryukyu Islands and the Izu Islands, those with 5-6 apertures were common.

Effects of nucleo-cytoplasmic interactions on leaf volatile compounds from citrus somatic diploid hybrids.

Three diploid citrus somatic hybrids (cybrids) were produced by fusions combining nucellar callus-derived protoplasts of Willow Leaf mandarin (Citrus deliciosa Ten.) and Commune clementine (Citrus clementina Hort. ex Tan.) with, respectively, leaf protoplasts of Eureka lemon [Citrus limon (L.) Burm.] and Marumi kumquat [Fortunella japonica (Thunb.) Swing.] and leaf protoplasts of Marumi kumquat. Ploidy and origins of the nuclear, chloroplastic, and mitochondrial genomes were investigated by flow cytometry and nuclear and cytoplasmic simple sequence repeat analyses. Volatile compounds were extracted from the leaves of the three cybrids by a pentane/ether (1:1) mixture, analyzed by GC-MS, and compared to those of their parents. The cybrids were found to be very close to their nucleus-giving parent, suggesting that the main information for volatile compounds biosynthesis is contained in the nucleus. However, nucleo-cytoplasmic interactions occurred: the (mandarin + lemon) cybrid, possessing nucleus and chloroplasts of lemon and mitochondria from mandarin, synthesizes more monoterpene alcohols and esters than its nucleus-giving parent; the (clementine + kumquat) cybrid, possessing nucleus from kumquat and organelles from mandarin, synthesizes more monoterpene and sesquiterpene hydrocarbons and sesquiterpene alcohols than its nucleus-giving parent.