Deep proteome analysis of gerontoplasts from the inner integument of developing seeds of Jatropha curcas.

UNLABELLED: The inner integument of Jatropha curcas seeds is a non-photosynthetic tissue that acts primarily as a conduit for the delivery of nutrients to the embryo and endosperm. In this study we performed a histological and transmission electron microscopy analysis of the inner integument in stages prior to fertilization to 25days after pollination, to establish the structural changes associated with the plastid to gerontoplast transition. This study showed that plastids are subjected to progressive changes, which include the dismantling of the internal membrane system, matrix degradation and the formation of stromule-derived vesicles. A proteome analysis of gerontoplasts isolated from the inner integument at 25days after pollination, resulted in the identification of 1923 proteins, which were involved in a myriad of metabolic functions, such as synthesis of amino acids and fatty acids. Among the identified proteins, were also a number of hydrolases (peptidases, lipases and carbohydrases), which presumably are involved in the ordered dismantling of this organelle to provide additional sources of nutrients for the growing embryo and endosperm. The dataset we provide here may provide a foundation for the study of the proteome changes associated with the plastid to gerontoplast transition in non-photosynthetic tissues. SIGNIFICANCE: We describe ultrastructural features of gerontoplasts isolated from the inner integument of developing seeds of Jatropha curcas, together with a deep proteome analysis of these gerontoplasts. This article explores a new aspect of the biology of plastids, namely the ultrastructural and proteome changes associated with the transition plastid to gerontoplast in a non-photosynthetic tissue.

Floral nectar production and nectary structure of a bee-pollinated shrub from Neotropical savanna.

Biotic pollination is critical for tropical ecosystem functioning, and nectar plays an essential role as it represents the main trophic resource for pollinators. Nevertheless, little is known about the mechanisms that underlie its production, which is essential for understanding the basis of nectar-mediated interactions in ecological and evolutionary approaches. Therefore, this study explores the relationship between the nectar secretion pattern and nectary functional changes in Anemopaegma album, a bee-pollinated species. We analysed the pattern of nectar production under field conditions and investigated floral nectary structural changes in two different developmental stages using light, transmission and scanning electron microscopy. We measured 30.95 ± 23.02 µl (mean ± SD, n = 30) of nectar accumulated inside the nectar chamber (29.26 ± 3.48% sucrose equivalents) at the moment of flower opening. Nectar removal did not influence the pattern of floral nectar production in terms of volume or total sugar but reduced the concentration of the nectar produced during the first 24 h of anthesis. The nectary consisted of an epidermis, a nectary parenchyma and a subnectary parenchyma supplied only by phloem. Starch grains decreased in size and abundance from the subnectary parenchyma toward the epidermis. We observed the degradation of starch grains and incorporation of amyloplasts into vacuoles at the pre-anthesis stage as well as the transformation of amyloplasts into elaioplasts during anthesis. Nectar secretion was continuous during the A. album flower life span, which was related to the functional features of its floral nectary, especially the presence of starch stored in the parenchyma.

Dissecting the mechanism of Solanum lycopersicum and Solanum chilense flower colour formation.

Flowers are the defining feature of angiosperms, and function as indispensable organs for sexual reproduction. Flower colour typically plays an important role in attracting pollinators, and can show considerable variation, even between closely related species. For example, domesticated tomato (S. lycopersicum) has orange/yellow flowers, while the wild relative S. chilense (accession LA2405) has bright yellow flowers. In this study, the mechanism of flower colour formation in these two species was compared by evaluating the accumulation of carotenoids, assessing the expression genes related to carotenoid biosynthetic pathways and observing chromoplast ultrastructure. In S. chilense petals, genes associated with the lutein branch of the carotenoid biosynthetic pathway, phytoene desaturase (PDS), ζ-carotene desaturase (ZDS), lycopene ß-cyclase (LCY-B), ß-ring hydroxylase (CRTR-B) and ε-ring hydroxylase (CRTR-E), were highly expressed, and this was correlated with high levels of lutein accumulation. In contrast, PDS, ZDS and CYC-B from the neoxanthin biosynthetic branch were highly expressed in S. lycopersicum anthers, leading to increased ß-carotene accumulation and hence an orange/yellow colour. Changes in the size, amount and electron density of plastoglobules in chromoplasts provided further evidence of carotenoid accumulation and flower colour formation. Taken together, these results reveal the biochemical basis of differences in carotenoid pigment accumulation and colour between petals and anthers in tomato.

Proteome analysis of plastids from developing seeds of Jatropha curcas L.

In this study, we performed a proteomic analysis of plastids isolated from the endosperm of developing Jatropha curcas seeds that were in the initial stage of deposition of protein and lipid reserves. Proteins extracted from the plastids were digested with trypsin, and the peptides were applied to an EASY-nano LC system coupled inline to an ESI-LTQ-Orbitrap Velos mass spectrometer, and this led to the identification of 1103 proteins representing 804 protein groups, of which 923 proteins were considered as true identifications, and this considerably expands the repertoire of J. curcas proteins identified so far. Of the identified proteins, only five are encoded in the plastid genome, and none of them are involved in photosynthesis, evidentiating the nonphotosynthetic nature of the isolated plastids. Homologues for 824 out of 923 identified proteins were present in PPDB, SUBA, or PlProt databases while homologues for 13 proteins were not found in any of the three plastid proteins databases but were marked as plastidial by at least one of the three prediction programs used. Functional classification showed that proteins belonging to amino acids metabolism comprise the main functional class, followed by carbohydrate, energy, and lipid metabolisms. The small and large subunits of Rubisco were identified, and their presence in the plastids is considered to be an adaptive feature counterbalancing for the loss of one-third of the carbon as CO2 as a result of the conversion of carbohydrate to oil through glycolysis. While several enzymes involved in the biosynthesis of several precursors of diterpenoids were identified, we were unable to identify any terpene synthase/cyclase, which suggests that the plastids isolated from the endosperm of developing seeds do not synthesize phorbol esters. In conclusion, our study provides insights into the major biosynthetic pathways and certain unique features of the plastids from the endosperm of developing seeds at the whole proteome level.

Ultrastructural features of Mimulus aurantiacus (Scrophulariaceae) pollen tubes in vivo.

The aim of this study is to give information on ultrastructure of in vivo pollen tubes of Mimulus aurantiacus which were collected from the Botanical Garden of the University of California at Berkeley. Materials were prepared according to electron microscopy methods and examined under Zeiss electron microscope. Four zones were examined in the pollen tubes of Mimulus aurantiacus. Apical zone: Mitochondria, smooth endoplasmic reticulum, rough endoplasmic reticulum, dictyosomes and secretory vesicles were observed. Subapical zone: This area contained abundant rough endoplasmic reticulum and occasionally some smooth endoplasmic reticulum. The polysomes, mitochondria, proplastids that contain starch, small vacuoles and a few lipid bodies were detected. Nuclear zone: Both generative and vegetative cell nuclei lie in this zone. The vegetative cell nucleus was large and long. Rough endoplasmic reticulum, mitochondria, ribosomes, dictyosomes, and amyloplasts that are rich of starch were observed. Vacuolation and plug formation zone: Cytoplasm of the tubes was full of large vacuoles. Few organelles such as mitochondria, dictyosome and rough endoplasmic reticulum were detected along their periphery.

Ultrastructural features of Mimulus aurantiacus (Scrophulariaceae) pollen tubes in vivo

The aim of this study is to give information on ultrastructure of in vivo pollen tubes of Mimulus aurantiacus which were collected from the Botanical Garden of the University of California at Berkeley. Materials were prepared according to electron microscopy methods and examined under Zeiss electron microscope. Four zones were examined in the pollen tubes of Mimulus aurantiacus. APICAL ZONE: Mitochondria, smooth endoplasmic reticulum, rough endoplasmic reticulum, dictyosomes and secretory vesicles were observed. SUBAPICAL ZONE: This area contained abundant rough endoplasmic reticulum and occasionally some smooth endoplasmic reticulum. The polysomes, mitochondria, proplastids that contain starch, small vacuoles and a few lipid bodies were detected. NUCLEAR ZONE: Both generative and vegetative cell nuclei lie in this zone. The vegetative cell nucleus was large and long. Rough endoplasmic reticulum, mitochondria, ribosomes, dictyosomes, and amyloplasts that are rich of starch were observed. VACUOLATION AND PLUG FORMATION ZONE: Cytoplasm of the tubes was full of large vacuoles. Few organelles such as mitochondria, dictyosome and rough endoplasmic reticulum were detected along their periphery.

O objetivo deste estudo é informar sobre a ultraestrutura de tubos de pólen de Mimulus aurantiacus in vivo coletados no "Botanical Garden" da Universidade da Califórnia em Berkeley. O material foi preparado de acordo com os métodos de microscopia eletrônica e examinado em microscópio eletrônico Zeiss. Quatro zonas dos tubos de pólen de Mimulus aurantiacus foram examinadas. ZONA APICAL: foram observados mitocôndrias, retículo endoplasmático liso; retículo endoplasmático rugoso, dictiossomos e vesículas secretoras. ZONA SUBAPICAL: esta área continha retículo endoplasmático rugoso em abundância e, ocasionalmente, algum retículo endoplasmático liso. Foram detectados polissomos, mitocôndrias, proplastídeos que contêm amido, pequenos vacúolos e alguns corpos lipídicos. ZONA NUCLEAR: nesta área, existem tanto núcleos de células geradoras como vegetativas. O núcleo de célula vegetativa é grande e longo. Foram observados retículo endoplasmático rugoso, mitocôndria, ribossomos, dictiossomos e amiloplastos ricos em amido. ZONA DE VACUOLIZAÇÃO E DE FORMAÇÃO DE "PLUG": o citoplasma dos tubos estava cheio de grandes vacúolos. Algumas organelas como mitocôndria, dictiossomo e retículo endoplasmático rugoso foram detectadas em toda a periferia desta área.

The floral nectary of Hymenaea stigonocarpa (Fabaceae, Caesalpinioideae): structural aspects during floral development.

BACKGROUND AND AIMS: Considering that few studies on nectary anatomy and ultrastructure are available for chiropterophilous flowers and the importance of Hymenaea stigonocarpa in natural 'cerrado' communities, the present study sought to analyse the structure and cellular modifications that take place within its nectaries during the different stages of floral development, with special emphasis on plastid dynamics. METHODS: For the structural and ultrastructural studies the nectary was processed as per usual techniques and studied under light, scanning and transmission electron microscopy. Histochemical tests were employed to identify the main metabolites on nectary tissue and secretion samples. KEY RESULTS: The floral nectary consists of the inner epidermis of the hypanthium and vascularized parenchyma. Some evidence indicates that the nectar release occurs via the stomata. The high populations of mitochondria, and their juxtaposition with amyloplasts, seem to be related to energy needs for starch hydrolysis. Among the alterations observed during the secretory phase, the reduction in the plastid stromatic density and starch grain size are highlighted. When the secretory stage begins, the plastid envelope disappears and a new membrane is formed, enclosing this region and giving rise to new vacuoles. After the secretory stage, cellular structures named 'extrastomatic bodies' were observed and seem to be related to the nectar resorption. CONCLUSIONS: Starch hydrolysis contributes to nectar formation, in addition to the photosynthates derived directly from the phloem. In these nectaries, the secretion is an energy-requiring process. During the secretion stage, some plastids show starch grain hydrolysis and membrane rupture, and it was observed that the region previously occupied by this organelle continued to be reasonably well defined, and gave rise to new vacuoles. The extrastomatic bodies appear to be related to the resorption of uncollected nectar.

Fine structural study of the red seaweed Gymnogongrus torulosus (Phyllophoraceae, Rhodophyta).

The present study analyzed several characters of the red seaweed Gymnogongrus torulosus, such as cellular structure of the thallus, cuticle, pit plug and cell wall ultrastructure, and morphology of some organelles like plastids, Golgi bodies and mitochondria. Also, anomalous chloroplasts with thylakoid disorganization were found in medullary cells. The significance of this thylakoid disposition is still unclear. This is one of the first studies focused on the fine structure of a red alga recorded in Argentina.

Fine structural study of the red seaweed Gymnogongrus torulosus (Phyllophoraceae, Rhodophyta)

The present study analyzed several characters of the red seaweed Gymnogongrus torulosus, such as cellular structure of the thallus, cuticle, pit plug and cell wall ultrastructure, and morphology of some organelles like plastids, Golgi bodies and mitochondria. Also, anomalous chloroplasts with thylakoid disorganization were found in medullary cells. The significance of this thylakoid disposition is still unclear. This is one of the first studies focused on the fine structure of a red alga recorded in Argentina.

Fine structural study of the red seaweed Gymnogongrus torulosus (Phyllophoraceae, Rhodophyta)

The present study analyzed several characters of the red seaweed Gymnogongrus torulosus, such as cellular structure of the thallus, cuticle, pit plug and cell wall ultrastructure, and morphology of some organelles like plastids, Golgi bodies and mitochondria. Also, anomalous chloroplasts with thylakoid disorganization were found in medullary cells. The significance of this thylakoid disposition is still unclear. This is one of the first studies focused on the fine structure of a red alga recorded in Argentina. (AU)