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Root-knot nematodes (RKNs) are plant pests that infect the roots of host plants. Bacillus thuringiensis (Bt) nematicidal proteins exhibited toxicity to nematodes. However, the application of nematicidal proteins for plant protection is hampered by the lack of effective delivery systems in transgenic plants. In this study, we discovered the accumulation of leucoplasts (root plastids) in galls and RKN-induced giant cells. RKN infection causes the degradation of leucoplasts into small vesicle-like structures, which are responsible for delivering proteins to RKNs, as observed through confocal microscopy and immunoelectron microscopy. We showed that different-sized proteins from leucoplasts could be taken up by Meloidogyne incognita female. To further explore the potential applications of leucoplasts, we introduced the Bt crystal protein Cry5Ba2 into tobacco and tomato leucoplasts by fusing it with a transit peptide. The transgenic plants showed significant resistance to RKNs. Intriguingly, RKN females preferentially took up Cry5Ba2 protein when delivered through plastids rather than the cytosol. The decrease in progeny was positively correlated with the delivery efficiency of the nematicidal protein. In conclusion, this study offers new insights into the feeding behavior of RKNs and their ability to ingest leucoplast proteins, and demonstrates that root leucoplasts can be used for delivering nematicidal proteins, thereby offering a promising approach for nematode control. This study uncovers that root-knot nematodes (RKNs) can absorb proteins from root leucoplasts. Based on this finding, an efficient system using root leucoplasts to deliver Bt nematicidal proteins is developed for controlling RKNs , offering a novel strategy for nematode control. [ABSTRACT FROM AUTHOR]

The ultrastructure of stigmatic and stylar secretory tissues was studied in one cultivar of Pyrus communis and six cultivars of Pyrus pyrifolia var. culta (the so-called Nashi pear) using standard light, scanning, and transmission electron microscopy methods. Although both tissues secreted an extracellular fluid necessary for the development of male gametophyte, they differed markedly in the ultrastructure of the extracellular matrix and in the distribution and ultrastructure of organelles. The difference was most evident in regard to the endoplasmic reticulum, which represented the rough, smooth, and vesicular type in stigmatic papillate epidermis and distal stigmatoid tissue cells and occurred mainly as the rough type in form of expanded cisternae filled with fine-fibrillar content in the transmitting tissue of the style.

The genus Spumella, established by Cienkowsky in 1870, is characterized by omnivory, two (rarely three) flagella, a short stick-like structure beneath the flagella, a threadlike stalk, cell division via constriction and cyst formation. Since the first phylogenetic study of Spumella-like flagellates, their paraphyly has consistently been shown, with separation into several genera. More recently, Spumella was carefully investigated using molecular and morphological data to propose seven new species. Classification of this genus and knowledge of its species diversity remain limited because Spumella-like flagellates are extremely difficult to identify based on limited morphological characters. To understand the phylogeny and taxonomy of Spumella, we analyzed molecular and morphological data from 47 strains, including 18 strains isolated from Korean ponds or swamps. Nuclear SSU, ITS and LSU rDNA data were used for maximum likelihood and Bayesian analyses. The molecular data divided the strains into 15 clades, including seven new lineages, each with unique molecular signatures for nuclear SSU rRNA from the E23-2 to E23-5 domains, the spacer between the E23-8 and E23-9 domains of the V4 region and domain 29 of the V5 region. Our results revealed increased species diversity in Spumella. In contrast to the molecular phylogeny results, the taxa showed very similar cell morphologies, suggesting morphological convergence into simple nanoflagellates to enable heterotrophy. Three new species produced stomatocysts in culture. Aspects of stomatocyst morphology, including collar structure, surface ornamentation, and cyst shape, were very useful in differentiating the three species. The general ultrastructure of Spumella bureschii strain Baekdongje012018B8 and S. benthica strain Hwarim032418A5 showed the typical chrysophyte form for the leucoplast, a vestigial chloroplast surrounded by four envelope membranes, supporting the hypothesis that Spumella evolved from a phototroph to a heterotroph via the loss of its photosynthetic ability. Seven new species are proposed: S. benthica, S. communis, S. longicolla, S. oblata, S. rotundata, S. similis, and S. sinechrysos.

The genus Spumella , established by Cienkowsky in 1870, is characterized by omnivory, two (rarely three) flagella, a short stick-like structure beneath the flagella, a threadlike stalk, cell division via constriction and cyst formation. Since the first phylogenetic study of Spumella -like flagellates, their paraphyly has consistently been shown, with separation into several genera. More recently, Spumella was carefully investigated using molecular and morphological data to propose seven new species. Classification of this genus and knowledge of its species diversity remain limited because Spumella -like flagellates are extremely difficult to identify based on limited morphological characters. To understand the phylogeny and taxonomy of Spumella , we analyzed molecular and morphological data from 47 strains, including 18 strains isolated from Korean ponds or swamps. Nuclear SSU, ITS and LSU rDNA data were used for maximum likelihood and Bayesian analyses. The molecular data divided the strains into 15 clades, including seven new lineages, each with unique molecular signatures for nuclear SSU rRNA from the E23-2 to E23-5 domains, the spacer between the E23-8 and E23-9 domains of the V4 region and domain 29 of the V5 region. Our results revealed increased species diversity in Spumella. In contrast to the molecular phylogeny results, the taxa showed very similar cell morphologies, suggesting morphological convergence into simple nanoflagellates to enable heterotrophy. Three new species produced stomatocysts in culture. Aspects of stomatocyst morphology, including collar structure, surface ornamentation, and cyst shape, were very useful in differentiating the three species. The general ultrastructure of Spumella bureschii strain Baekdongje012018B8 and S. benthica strain Hwarim032418A5 showed the typical chrysophyte form for the leucoplast, a vestigial chloroplast surrounded by four envelope membranes, supporting the hypothesis that Spumella evolved from a phototroph to a heterotroph via the loss of its photosynthetic ability. Seven new species are proposed: S. benthica , S. communis , S. longicolla , S. oblata , S. rotundata , S. similis , and S. sinechrysos. [ABSTRACT FROM AUTHOR]

Plastids are pivotal subcellular organelles that have evolved to perform specialized functions in plant cells, including photosynthesis and the production and storage of metabolites. They come in a variety of forms with different characteristics, enabling them to function in a diverse array of organ/tissue/cell-specific developmental processes and with a variety of environmental signals. Here, we have comprehensively reviewed the distinctive roles of plastids and their transition statuses, according to their features. Furthermore, the most recent understanding of their regulatory mechanisms is highlighted at both transcriptional and post-translational levels, with a focus on the greening and non-greening phenotypes.

Spumella-like heterotrophic chrysophytes are important eukaryotic microorganisms that feed on bacteria in aquatic and soil environments. They are characterized by their lack of pigmentation, naked cell surface, and extremely small size. Although Spumella-like chrysophytes have lost their photosynthetic ability, they still possess a leucoplast and retain a plastid genome. We have sequenced the plastid genomes of three non-photosynthetic chrysophytes, Spumella sp. Baeckdong012018B8, Pedospumella sp. Jangsampo120217C5 and Poteriospumella lacustris Yongseonkyo072317C3, and compared them to the previously sequenced plastid genome of “Spumella” sp. NIES-1846 and photosynthetic chrysophytes. We found the plastid genomes of Spumella-like flagellates to be generally conserved with respect to genome structure and housekeeping gene content. We nevertheless also observed lineage-specific gene rearrangements and duplication of partial gene fragments at the boundary of the inverted repeat and single copy regions. Most gene losses correspond to genes for proteins involved in photosynthesis and carbon fixation, except in the case of petF. The newly sequenced plastid genomes range from ~55.7 kbp to ~62.9 kbp in size and share a core set of 45 protein-coding genes, 3 rRNAs, and 32 to 34 tRNAs. Our results provide insight into the evolutionary history of organelle genomes via genome reduction and gene loss related to loss of photosynthesis in chrysophyte evolution.

It is unclear whether light affects the structure and activity of exogenous secretory tissues like glandular hairs. Therefore, transmission electron microscopy was first used to study plastid differentiation in glandular hairs and leaves of light-grown rosemary (Rosmarinus officinalis “Arp”) plants kept for 2 weeks under ambient light conditions. During our detailed analyses, among others, we found leucoplasts with tubuloreticular membrane structures resembling prolamellar bodies in stalk cell plastids of peltate glandular hairs. To study the effect of darkness on plastid differentiation, we then dark-forced adult, light-grown rosemary plants for 2 weeks and observed occasionally the development of new shoots with elongated internodes and pale leaves on them. Absorption and fluorescence spectroscopic analyses of the chlorophyllous pigment contents, the native arrangement of the pigment-protein complexes and photosynthetic activity confirmed that the first and second pairs of leaf primordia of dark-forced shoots were partially etiolated (contained low amounts of protochlorophyll/ide and residual chlorophylls, had etio-chloroplasts with prolamellar bodies and low grana, and impaired photosynthesis). Darkness did not influence plastid structure in fifth leaves or secretory tissues (except for head cells of peltate glandular hairs in which rarely tubuloreticular membranes appeared). The mesophyll cells of cotyledons of 2-week-old dark-germinated rosemary seedlings contained etioplasts with highly regular prolamellar bodies similar to those in mesophyll etio-chloroplasts of leaves and clearly differing from tubuloreticular membranes of secretory cells. Analyses of the essential oil composition obtained after solid phase microextraction and gas chromatography-mass spectroscopy showed that in addition to light, the age of the studied organ (i.e., first leaf primordia and leaf tip vs. fifth, fully developed green leaves) and the type of the organ (cotyledon vs. leaves) also strongly influenced the essential oil composition. Therefore, light conditions and developmental stage are both important factors to be considered in case of potential therapeutic, culinary or aromatic uses of rosemary leaves and their essential oils.

Spumella -like heterotrophic chrysophytes are important eukaryotic microorganisms that feed on bacteria in aquatic and soil environments. They are characterized by their lack of pigmentation, naked cell surface, and extremely small size. Although Spumella -like chrysophytes have lost their photosynthetic ability, they still possess a leucoplast and retain a plastid genome. We have sequenced the plastid genomes of three non-photosynthetic chrysophytes, Spumella sp. Baeckdong012018B8, Pedospumella sp. Jangsampo120217C5 and Poteriospumella lacustris Yongseonkyo072317C3, and compared them to the previously sequenced plastid genome of " Spumella " sp. NIES-1846 and photosynthetic chrysophytes. We found the plastid genomes of Spumella -like flagellates to be generally conserved with respect to genome structure and housekeeping gene content. We nevertheless also observed lineage-specific gene rearrangements and duplication of partial gene fragments at the boundary of the inverted repeat and single copy regions. Most gene losses correspond to genes for proteins involved in photosynthesis and carbon fixation, except in the case of pet F. The newly sequenced plastid genomes range from ~55.7 kbp to ~62.9 kbp in size and share a core set of 45 protein-coding genes, 3 rRNAs, and 32 to 34 tRNAs. Our results provide insight into the evolutionary history of organelle genomes via genome reduction and gene loss related to loss of photosynthesis in chrysophyte evolution. [ABSTRACT FROM AUTHOR]

It is unclear whether light affects the structure and activity of exogenous secretory tissues like glandular hairs. Therefore, transmission electron microscopy was first used to study plastid differentiation in glandular hairs and leaves of light-grown rosemary (Rosmarinus officinalis "Arp") plants kept for 2 weeks under ambient light conditions. During our detailed analyses, among others, we found leucoplasts with tubuloreticular membrane structures resembling prolamellar bodies in stalk cell plastids of peltate glandular hairs. To study the effect of darkness on plastid differentiation, we then dark-forced adult, light-grown rosemary plants for 2 weeks and observed occasionally the development of new shoots with elongated internodes and pale leaves on them. Absorption and fluorescence spectroscopic analyses of the chlorophyllous pigment contents, the native arrangement of the pigment-protein complexes and photosynthetic activity confirmed that the first and second pairs of leaf primordia of dark-forced shoots were partially etiolated (contained low amounts of protochlorophyll/ide and residual chlorophylls, had etio-chloroplasts with prolamellar bodies and low grana, and impaired photosynthesis). Darkness did not influence plastid structure in fifth leaves or secretory tissues (except for head cells of peltate glandular hairs in which rarely tubuloreticular membranes appeared). The mesophyll cells of cotyledons of 2-week-old dark-germinated rosemary seedlings contained etioplasts with highly regular prolamellar bodies similar to those in mesophyll etio-chloroplasts of leaves and clearly differing from tubuloreticular membranes of secretory cells. Analyses of the essential oil composition obtained after solid phase microextraction and gas chromatography-mass spectroscopy showed that in addition to light, the age of the studied organ (i.e., first leaf primordia and leaf tip vs. fifth, fully developed green leaves) and the type of the organ (cotyledon vs. leaves) also strongly influenced the essential oil composition. Therefore, light conditions and developmental stage are both important factors to be considered in case of potential therapeutic, culinary or aromatic uses of rosemary leaves and their essential oils. [ABSTRACT FROM AUTHOR]

Recently, a recessive Arabidopsis thaliana mutant with abundant stromules in leaf epidermal pavement cells was visually screened and isolated. The gene responsible for this mutant phenotype was identified as PARC6, a chloroplast division site regulator gene. The mutant allele parc6-5 carried two point mutations (G62R and W700stop) at the N- and C-terminal ends of the coding sequence, respectively. Here, we further characterized parc6-5 and other parc6 mutant alleles, and showed that PARC6 plays a critical role in plastid morphogenesis in all cell types of the leaf epidermis: pavement cells, trichome cells, and guard cells. Transient expression of PARC6 transit peptide (TP) fused to the green fluorescent protein (GFP) in plant cells showed that the G62R mutation has no or little effect on the TP activity of the PARC6 N-terminal region. Then, plastid morphology was microscopically analyzed in the leaf epidermis of wild-type (WT) and parc6 mutants (parc6-1, parc6-3, parc6-4 and parc6-5) with the aid of stroma-targeted fluorescent proteins. In parc6 pavement cells, plastids often assumed aberrant grape-like morphology, similar to those in severe plastid division mutants, atminE1, and arc6. In parc6 trichome cells, plastids exhibited extreme grape-like aggregations, without the production of giant plastids (>6 µm diameter), as a general phenotype. In parc6 guard cells, plastids exhibited a variety of abnormal phenotypes, including reduced number, enlarged size, and activated stromules, similar to those in atminE1 and arc6 guard cells. Nevertheless, unlike atminE1 and arc6, parc6 exhibited a low number of mini-chloroplasts (< 2 µm diameter) and rarely produced chloroplast-deficient guard cells. Importantly, unlike parc6, the chloroplast division site mutant arc11 exhibited WT-like plastid phenotypes in trichome and guard cells. Finally, observation of parc6 complementation lines expressing a functional PARC6-GFP protein indicated that PARC6-GFP formed a ring-like structure in both constricting and non-constricting chloroplasts, and that PARC6 dynamically changes its configuration during the process of chloroplast division.

Recently, a recessive Arabidopsis thaliana mutant with abundant stromules in leaf epidermal pavement cells was visually screened and isolated. The gene responsible for this mutant phenotype was identified as PARC6 , a chloroplast division site regulator gene. The mutant allele parc6-5 carried two point mutations (G62R and W700stop) at the N- and C-terminal ends of the coding sequence, respectively. Here, we further characterized parc6-5 and other parc6 mutant alleles, and showed that PARC6 plays a critical role in plastid morphogenesis in all cell types of the leaf epidermis: pavement cells, trichome cells, and guard cells. Transient expression of PARC6 transit peptide (TP) fused to the green fluorescent protein (GFP) in plant cells showed that the G62R mutation has no or little effect on the TP activity of the PARC6 N-terminal region. Then, plastid morphology was microscopically analyzed in the leaf epidermis of wild-type (WT) and parc6 mutants (parc6-1 , parc6-3 , parc6-4 and parc6-5) with the aid of stroma-targeted fluorescent proteins. In parc6 pavement cells, plastids often assumed aberrant grape-like morphology, similar to those in severe plastid division mutants, atminE1, and arc6. In parc6 trichome cells, plastids exhibited extreme grape-like aggregations, without the production of giant plastids (>6 µm diameter), as a general phenotype. In parc6 guard cells, plastids exhibited a variety of abnormal phenotypes, including reduced number, enlarged size, and activated stromules, similar to those in atminE1 and arc6 guard cells. Nevertheless, unlike atminE1 and arc6 , parc6 exhibited a low number of mini-chloroplasts (< 2 µm diameter) and rarely produced chloroplast-deficient guard cells. Importantly, unlike parc6 , the chloroplast division site mutant arc11 exhibited WT-like plastid phenotypes in trichome and guard cells. Finally, observation of parc6 complementation lines expressing a functional PARC6-GFP protein indicated that PARC6-GFP formed a ring-like structure in both constricting and non-constricting chloroplasts, and that PARC6 dynamically changes its configuration during the process of chloroplast division. [ABSTRACT FROM AUTHOR]

The role of glutathione in the plant vacuole is still being debated. In the present paper, the redox state of glutathione and the activity of glutathione S-transferase (GST, E 2.5.1.18) in the vacuole compared to those in leucoplast have been studied. Organelles were isolated from dormant red beet (Beta vulgaris L.) taproots. Two generally used approaches have been applied to quantitatively assess the content of glutathione. Initially, levels of glutathione were measured in isolated organelles after labeling with monochlorobimane (MCB) and imaging with the use of confocal laser scanning microscopy. However, there are factors limiting the specificity of this method, because of which the resulting concentrations of vacuolar GSH have been underestimated. Another approach used was HPLC, which allows to simultaneously quantify the reduced glutathione (GSH) and glutathione disulfide (GSSG). The concentration of the total glutathione (GSHt) and GSSG in vacuoles determined with the aid of HPLC-UV was higher in comparison to that in the leucoplasts. The reduction potential (E h) for the glutathione couple in the vacuoles was more positive (−163 mV), than that in plastids (−282 mV). The relatively rapid increase in fluorescence in the isolated vacuoles and plastids during MCB-labeling has indicated to the contribution of GSTs, since the conjugation of GSH to bimane is catalysed by these enzymes. The GST activity in the vacuoles has been assessed to be quite high compared to that of leucoplasts. The number of isoforms of GSTs also differed markedly in vacuoles and plastids. Collectively, our findings suggest the idea that the glutathione accumulated by central vacuole seems to contribute to the redox processes and to the detoxification, which can take place in this compartment. • The levels of the total glutathione (GSHt) in vacuoles are relatively high with respect to those in leucoplasts during. • GSSG accounts for more than 20% of GSHt content in vacuoles and 8% of GSHt content in leucoplasts. • E h for the glutathione couple of vacuoles was not so reducing like that for leucoplasts. • Glutathione S-transferases (GSTs) function in vacuoles. • The activity of GSTs and their isoenzyme composition differed markedly in vacuoles versus leucoplasts. [ABSTRACT FROM AUTHOR]

Plastids represent a largely diverse group of organelles in plant and algal cells that have several common features but also a broad spectrum of morphological, ultrastructural, biochemical, and physiological differences. Plastids and their structural and metabolic diversity significantly contribute to the functionality and developmental flexibility of the plant body throughout its lifetime. In addition to the multiple roles of given plastid types, this diversity is accomplished in some cases by interconversions between different plastids as a consequence of developmental and environmental signals that regulate plastid differentiation and specialization. In addition to basic plastid structural features, the most important plastid types, the newly characterized peculiar plastids, and future perspectives in plastid biology are also provided in this chapter., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

The plastid proteome changes according to developmental stages. Accruing evidence shows that, in addition to transcriptional and translational controls, preprotein import into plastids is also part of the process regulating plastid proteomes. Different preproteins have distinct preferences for plastids of different tissues. Preproteins are also divided into at least three age-selective groups based on their import preference for chloroplasts of different ages. Both tissue and age selectivity are determined by the transit peptide of each preprotein, and a transit-peptide motif for older-chloroplast preference has been identified. Future challenges lie in identifying other motifs for tissue and age selectivity, as well as in identifying the receptor components that decipher these motifs. Developmental regulation also suggests that caution should be exercised when comparing protein import data generated with plastids isolated from different tissues or with chloroplasts isolated from plants of different ages. [ABSTRACT FROM AUTHOR]

Micromorphological investigations on the terpene-producing trichomes at the active secretory stage were performed on leaves and flowers of the Mexican Salvia greggii A. Gray (Lamiaceae), cultivated in Italy. Two types of glandular trichomes are described and compared: peltate, widespread on both the vegetative and the reproductive organs, and medium-stalked capitate, typical of the calyx abaxial side. The histochemical evidences indicate an exclusive terpene production for both morphotypes. TEM survey highlighted several common ultrastructural features that validated the light microscope observations, however a great variability was detected at the plastidome level. Different plastid populations characterized each trichome type, also in relation to the different distribution pattern on the plant epidermis. In the peltates plastids invariably displayed a well-developed internal membrane system and various types of osmiophilic inclusions: on the leaf and the calyx plastoglobuli were also observed, whereas on the corolla abundant starch grains. True leucoplast are only detected in the calyx capitate trichomes. 49 compounds were detected in the essential oil (EO) obtained from the aerial parts, with terpenes representing 98.76% of the total EO, besides negligible amounts of non-terpene derivatives and apocarotenoids. Sesquiterpenes and monoterpenes occurred in similar relative abundances: 52.7% versus 46.1%, respectively. Among sesquiterpenes, γ-muurolene (11.48%) and guaiol (6.15%) dominated for hydrocarbons and oxygenated compounds, respectively. β-pinene (20.24%) prevailed among the monoterpene hydrocarbons, resulting also as the most abundant compound in the essential oil. 1,8-cineole (11.00%) is the dominant compound of the oxygenated monoterpenes. The ultrastructural appearance of the plastidome was linked with the various terpene classes present in the EO profile. We argued that monoterpenes were probably synthetized in the calyx capitate trichomes and the sesquiterpenes in the peltates. [ABSTRACT FROM AUTHOR]

Plastoglobuli (PGs) are plastid lipoprotein particles surrounded by a membrane lipid monolayer. PGs contain small specialized proteomes and metabolomes. They are present in different plastid types (e.g., chloroplasts, chromoplasts, and elaioplasts) and are dynamic in size and shape in response to abiotic stress or developmental transitions. PGs in chromoplasts are highly enriched in carotenoid esters and enzymes involved in carotenoid metabolism. PGs in chloroplasts are associated with thylakoids and contain ∼30 core proteins (including six ABC1 kinases) as well as additional proteins recruited under specific conditions. Systems analysis has suggested that chloroplast PGs function in metabolism of prenyl lipids (e.g., tocopherols, plastoquinone, and phylloquinone); redox and photosynthetic regulation; plastid biogenesis; and senescence, including recycling of phytol, remobilization of thylakoid lipids, and metabolism of jasmonate. These functionalities contribute to chloroplast PGs' role in responses to stresses such as high light and nitrogen starvation. PGs are thus lipid microcompartments with multiple functions integrated into plastid metabolism, developmental transitions, and environmental adaptation. This review provides an in-depth overview of PG experimental observations, summarizes the present understanding of PG features and functions, and provides a conceptual framework for PG research and the realization of opportunities for crop improvement. [ABSTRACT FROM AUTHOR]

Premise of research. The genus Citrus includes species that are among the most important tree fruit crops. Although fruit juice is the primary product, monoterpenoid essential oil obtained from the peel is an important value-added commodity. Peel monoterpenes are synthesized by subepidermal secretory cavities that consist of glandular cells surrounding an extracellular oil storage space. Several previous studies have focused on early secretory cavity development, ending with the initiation of secretion. In order to better understand the process of monoterpene formation, transport, and storage, it is important to obtain detailed information concerning plant oil glands during all phases of development. Methodology. TEM was performed on secretory cavities isolated from the exocarp of grapefruit (Citrus x paradisi Macfad. 'Duncan') preserved by microwave-assisted fixation or high-pressure freezing and freeze substitution. Immunocytochemistry was employed to localize (+)-limonene synthase. Tomography was used to examine leucoplast ultrastructure. Pivotal results. Glandular epithelial cells of secretory cavities persisted for several months after their formation. Lipid accumulation occurred within plastoglobule-like lipid bodies (PGs), vacuoles, and cytoplasmic lipid bodies of epithelial cells. (+)-Limonene synthase, the enzyme that catalyzes the primary flux-carrying reaction in Citrus monoterpene biosynthesis, was localized to tubules within oil gland leucoplasts and was not associated with PGs. No bulk movement of lipids from PGs to vacuoles or to the extracellular oil storage cavity was observed. Conclusions. Oil glands remain biosynthetically active throughout fruit development in grapefruit. The high degree of oil gland metabolic specialization is reflected in unique cellular features, such as leucoplasts, abundant plastid-endoplasmic reticulum membrane contact sites, and the accumulation of fibrillar material in vacuoles. The mechanism of the movement of monoterpenes from plastids to the central cavity is unknown, but our observations suggest that it does not involve vesicles or disruption of the large PGs. [ABSTRACT FROM AUTHOR]

The behaviour and multiplication of pollen plastids have remained elusive despite their crucial involvement in cytoplasmic inheritance. Here, we present live images of plastids in pollen grains and growing tubes from transgenic Arabidopsis thaliana lines expressing stroma-localised FtsZ1–green-fluorescent protein fusion in a vegetative cell-specific manner. Vegetative cells in mature pollen contained a morphologically heterogeneous population of round to ellipsoidal plastids, whilst those in late-developing (maturing) pollen included plastids that could have one or two constriction sites. Furthermore, plastids in pollen tubes exhibited remarkable tubulation, stromule (stroma-filled tubule) extension, and back-and-forth movement along the direction of tube growth. Plastid division, which involves the FtsZ1 ring, was rarely observed in mature pollen grains. [ABSTRACT FROM AUTHOR]

Stromules (stroma-filled tubules) are highly dynamic structures extending from the surface of all plastid types examined so far, including proplastids, chloroplasts, etioplasts, leucoplasts, amyloplasts, and chromoplasts. Stromules are usually 0.35–0.85 μm in diameter and of variable length, from short beak-like projections to linear or branched structures up to 220 μm long. They are enclosed by the inner and outer plastid envelope membranes and enable the transfer of molecules as large as Rubisco (∼560 kDa) between interconnected plastids. Stromules occur in all cell types, but stromule morphology and the proportion of plastids with stromules vary from tissue to tissue and at different stages of plant development. In general, stromules are more abundant in tissues containing non-green plastids, and in cells containing smaller plastids. The primary function of stromules is still unresolved, although the presence of stromules markedly increases the plastid surface area, potentially increasing transport to and from the cytosol. Other functions of stromules, such as transfer of macromolecules between plastids and starch granule formation in cereal endosperm, may be restricted to particular tissues and cell types. [ABSTRACT FROM PUBLISHER]

A cDNA encoding NADP-dependent malic enzyme was isolated from castor bean (Ricinus communis L.) endosperm. The 2362-bp cDNA encodes a 641 amino acid protein, which is predicted to have a 50 amino acid chloroplast transit peptide. Southern blotting revealed a single copy of the gene. Recombinant protein expression confirmed that the cDNA encodes a NADP-dependent malic enzyme; both Western blotting and enzyme assays detected NADP-dependent malic enzyme. Removal of the transit peptide in the recombinant protein did not affect the pH optimum of the enzyme, but it did result in an increased specific activity. [ABSTRACT FROM AUTHOR]

Summary: Two heterotrophic members of the Dictyochophyceae (stramenopiles), Pteridomonas danica and Ciliophrys infusionum, were investigated. An undescribed organelle bounded by four membranes and closely associated with the nucleus was detected in P. danica. The outermost membrane was continuous with the outer nuclear membrane. These features strongly suggested that this organelle was a vestigial chloroplast. A photosynthetic gene, rbc L, was successfully amplified by polymerase chain reaction (PCR) from P. danica and C. infusionum. These sequences were readily and well aligned with those of photosynthetic stramenopiles. Phylogenetic trees of 18S rDNA and rbc L were constructed. In all the trees obtained, P. danica and C. infusionum appeared in two different clades, the Pedinellales clade and the Ciliophryales/Rhizochromulinales clade, each of which contained photosynthetic members as well as heterotrophic members. The results indicated that the loss of photosynthetic ability occurred independently in P. danica and C. infusionum. This is the first report of the presence of a vestigial chloroplast (leucoplast) in colorless dictyochophytes. [Copyright &y& Elsevier]

The subcellular compartmentation of isopentenyl diphosphate (IPP) synthesis was examined in secretory cells isolated from glandular trichomes of peppermint ( Mentha x piperita L. cv. Black Mitcham). As a consequence of their anatomy and the conditions of their isolation, the isolated secretory cells are non-specifically permeable to low-molecular-weight water-soluble metabolites. Thus, the cytoplasm is readily accessible to the exogenous buffer whereas the selective permeability of subcellular organelles is maintained. With the appropriate choice of exogenous substrates, this feature allows the assessment of cytoplasmic and organellar (e.g. plastidic) metabolism in situ. Glycolytic substrates such as [C]glucose-6-phosphate and [C]pyruvic acid are incorporated into both monoterpenes and sesquiterpenes with a monoterpene:sesquiterpene ratio that closely mimics that observed in vivo, indicating that the correct subcellular partitioning of these substrates is maintained in this model system. Additionally, exogenous [C]mevalonic acid and [C]IPP, which are both intitially metabolized in the cytoplasm, produce an abnormally high proportion of sesquiterpenes. In contrast, incubation with either [C]citrate or [C]acetyl-CoA results in the accumulation of 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) with no detectable isoprenoids formed. Taken together, these results indicate that the cytoplasmic mevalonic acid pathway is blocked at HMG-CoA reductase and that the IPP utilized for both monoterpene and sesquiterpene biosynthesis is synthesized exclusively in the plastids. [ABSTRACT FROM AUTHOR]

The greening of the upper part of the outer Allium cepa L. bulb scales, in particular along the vascular regions, is limited to the hypodermal cells in which typical leucoplasts are transformed to normal and functional chloroplasts. This process is light dependent and cannot afterwards be reversed or modified by darkness. The changes in fine structure are described and briefly discussed. [ABSTRACT FROM AUTHOR]

Quantitative electron microscopy of serial sections was used to study the Polytoma papillatum cell and some of its constituents (nucleus, chondriome, leucoplast) during its vegetative life cycle. The volumes of cells just entering into or passing through mitosis varied considerably and seemed to determine the number of subsequent division processes. Whereas a volumetric balance existed between the cell (100%) and the chondriome (8-9%) during the whole life cycle, there was a correlation between cell and nuclear volumes (8-10%) only during interphase growth and the onset of mitosis. At telophase the nucleus-to-cell-volume ratio was reduced to 2%, but gradually increased during cytokinesis (4.6% at early cytokinesis; 6.5% at late cytokinesis) until it reached the initial value again in newly formed daughter cells. The leucoplast-to-cell-volume ratio (10-26%) varied considerably without any recognizable dependence upon cell cycling. The mean short axis of mitochondrial profiles was proportional to the mean diameter (=thickness) of the mitochondria; the specific surface (outer membrane area per 100 μm mitochondrial volume), and the surface-to-volume ratio changed rhythmically. Changes in mitochondrial surface-to-volume ratio (Sc/Vc) were apparently correlated with changes in mitochondrial diameter (Dc). This relationship can be approximately described by the function Sc/Vc=4/Dc. Deviations of the surface-to-volume ratios of the nuclei from the surface-to-volume ratios of idealized spheres of equal size, indicating profound changes in nuclear shape, were found mainly during mitosis. Results were compared with those obtained from other morphometric investigations and discussed with regard to their functional meaning. [ABSTRACT FROM AUTHOR]

The development of proplastids to chloroplasts or leucoplasts was followed in uniseriate nonglandular hairs, containing chloroplasts, and in glandular hairs, containing leucoplasts, of Stachys lanata leaves. Both hairs grow at the same time from the epidermis. Both chloroplasts and leucoplasts originate from structurally identical proplastids in meristematic cells. At the early stage, ribosomes are absent. After production of the first ribosomes, each plastid population differentiates along its own developmental pathway. In nonglandular hairs, ribosome multiplication occurs within the plastid stroma and a thylakoid system with grana stacks is formed. Ribosomes are attached to membrane vesicles produced by the inner membrane of the plastid envelope, to thylakoids, or are clustered in specialized areas of the stroma. Further development produces typical, small differentiated chloroplasts. In glandular hairs, the production of additional complete ribosomes ceases early. Smaller, transient dense particles are seen within the stroma in the course of the developmental process. The plastids increase in size and there is a transient tubular system but no thylakoids develop. Plastid division proceeds by neck constriction. The latter stage of leucoplast differentiation is characterized by the complete disappearance of ribosomal material and of tubular inner membranes. DNA nucleoids are obvious throughout the developmental period. Unlike chloroplasts, the development of these nonphotosynthetic heterotrophic organelles and the expression of their specialized metabolic activity probably does not involve the synthesis of specific peptides via plastoribosomes. [ABSTRACT FROM AUTHOR]