Was the evolution of plastid genetic machinery discontinuous?

The plastid nucleoid consists of plastid DNA and various, mostly uncharacterized, DNA-binding proteins. The plastid DNA undoubtedly originated from an ancestral cyanobacterial genome, but the origin of the nucleoid proteins appears complex. Initial biochemical analysis of these proteins, as well as comparative genome informatics, suggest that proteins of eukaryotic origin replaced most of the original prokaryotic proteins during the evolution of plastids in the lineage of green plants.     

Analysis of the 5′-leader regions of several plastid genes in protozoa of the phylum apicomplexa and red algae

Apicomplexan parasites contain so-called apicoplasts, which are similar to chloroplasts of red algae. Multiple alignments of the 5′-leader regions of plastid-encoded genes revealed several conserved noncoding regions in parasites as well as in red algae. The regions were assumed to be sites for RNA interactions with regulatory proteins. Conserved sites were found upstream of ycf24, which is required for [Fe-S] cluster development, and several other genes. In particular, a simultaneous regulation was predicted for ycf24, rps4, and rpo B in Toxoplasma gondii. The prediction agreed with the known data that apicoplasts are only required for a short time, but confer pathogenicity on T. gondii. Another site was predicted upstream of rpo B, which encodes the β subunit of RNA polymerase, in red algae Porphyra spp. and parasites Eimeria tenella and Theileria parva.     

Phylogeny of the tribe Colletieae (Rhamnaceae) – a sensitivity analysis of the plastid region trnL-trnF combined with morphology

The phylogenetic relationships within the tribe Colletieae (Rhamnaceae) were examined combining data from a previous morphological analysis with data from the trnL intron and trnL-F spacer. Previous studies have failed to confirm monophyly of the genus Discaria, the only genus of the tribe with an amphiantarctic distribution. The data set was analyzed using direct optimization as implemented in the computer program POY. Direct optimization searches for multiple optimal sequence alignments and is therefore well suited for analyzing DNA sequences including ambiguous alignable regions as found in the present study. Eight different costs were used for treating the indel information. Indels were treated as single events, equal to a fifth character state, or strings of gaps were treated as single events using different costs for opening a gap and extending the gap. The optimal cost set was selected by use of both character-based and topological congruence measures. Both congruence measures agreed upon a single optimal cost set. The resulting tree generally agrees with the current taxonomic treatment of the tribe Colletieae that recognizes six genera out of which three are monotypic. However, monophyly of Discaria was not supported and the results strongly suggest segregating D. nana and D. trinervis, and re-establishing the genus Ochetophila.We are grateful to the curator of the Herbario Gaspar Xuárez, Universidad de Buenos Aires (BAA), and to the curators of the Botanic Garden of the University of Copenhagen, Botanischer Garten und Botanisches Museum Berlin-Dahlem, Jardín Botánico Lucien Hauman (Facultad de Agronomía de la Universidad de Buenos Aires), Rancho Santa Ana Botanic Garden, and Royal Botanic Gardens, Melbourne, for permission to use material from their collections. María Elena Arce, María Martha Bianchi, Eugenia Chaia, and Neville Walsh kindly provided freshly collected material. We also thank Pablo A. Goloboff, Niels Klazenga, Pauline Y. Ladiges, Martín Ramírez, Ole Seberg, Llywela Williams, and two anonymous reviewers for critically reading earlier drafts of this paper. This work was supported in part by grants TG 028 (Universidad de Buenos Aires) and PIP 4027/96 (CONICET) to D. Medan. The University of Copenhagen supported Lone Aagesen (Ph.D. grant). Jürgen Kellermann received a Ph.D.-scholarship from the University of Melbourne.     

Changes in the protein and activity levels of the plastid NADH–plastoquinone–oxidoreductase complex during fruit development

Plastids contain an NADH dehydrogenase complex (Ndh complex) homologous to the mitochondrial complex I (EC 1.6.5.3). In this work, we have analysed the changes in the Ndh complex during ripening of pepper (Capsicum annum L., cv. Maor) and tomato (Lycopersicon esculentum Mill., cv. Marglobe) fruits. The Ndh complex was mainly present in the outer pericarp of tomato fruits, whereas it was evenly distributed in the pericarp of pepper. In both kinds of fruit we observed a decrease in the total amount of Ndh complex from the green to the red stage of development. This decrease corresponds to parallel decreases in the content and activity of the complex in plastids during the transition from chloroplasts to chromoplasts. Levels of plastidial quinol peroxidase activity were also higher during the first stages of tomato fruit development than during the latter stages of ripening. However, when referred to total plastid protein, the amount and activity of the Ndh complex in chloroplasts isolated from green fruits was higher than in chloroplasts isolated from leaves. These results strongly suggest that function of the Ndh complex, probably related to a plastidial electron transport chain, can be important during the first stages of fruit development.     

Erwin Baur or Carl Correns: who really created the theory of plastid inheritance?

Historical reviews of the field of non-Mendelian genetics and many other publications credit Erwin Baur and Carl Correns equally for the development of the theory of plastid inheritance. However, a study of the original literature indicates that this conclusion is not correct. Analysis of the relevant articles leads to the conclusion that Baur alone deserves credit for the theory of plastid inheritance. In his classic article on the inheritance properties of white-margined Pelargonium plants, Baur (1909) stated: (1) The plastids are carriers of hereditary factors which are able to mutate. (2) In variegated plants, random sorting-out of plastids is taking place. (3) The genetic results indicate a biparental inheritance of plastids by egg cells and sperm cells in Pelargonium. By contrast, Correns held the view that in variegated plants there is a maternally transmitted labile state of the cytoplasm which switches either to a permanently "healthy" state (allowing the "indifferent" plastids to become green chloroplasts) or to a permanently "diseased, ill" cytoplasmic state (causing white plastids and cells). Otto Renner supported Baur's theory and worked out important characteristics of plastid inheritance in the genus Oenothera. In the 1930s Renner reported many more observations, which established plastid inheritance as a widely accepted genetic theory.     

Solubilization of the plastidial lysophosphatidylcholine acyltransferase from Allium porrum leaves: towards plants devoid of eukaryotic plastid lipids?

To analyse the involvement of the plastidial lysophosphatidylcholine (lyso-PC) acyltransferase in the import of the extraplastidial lipid precursors required for eukaryotic plastid lipid synthesis, we plan to obtain transgenic plants. Since no sequence of lyso-PC acyltransferase is known, the purification of this enzyme has been undertaken to establish its sequence. First we determined the conditions allowing the solubilization of this membrane-bound enzyme. It is shown that by using CHAPS as a detergent, a lyso-PC acyltransferase activity is associated with the solubilized proteins.     

OsCpn60α1, encoding the plastid chaperonin 60α subunit, is essential for folding of rbcL

Chaperonins are involved in protein-folding. The rice genome encodes six plastid chaperonin subunits (Cpn60) — three α and three β. Our study showed that they were differentially expressed during normal plant development. Moreover, five were induced by heat stress (42°C) but not by cold (10°C). The oscpn60α1 mutant had a pale-green phenotype at the seedling stage and development ceased after the fourth leaf appeared. Transiently expressed OsCpn60α1:GFP fusion protein was localized to the chloroplast stroma. Immuno-blot analysis indicated that the level of Rubisco large subunit (rbcL) was severely reduced in the mutant while levels were unchanged for some imported proteins, e.g., stromal heat shock protein 70 (Hsp70) and chlorophyll a/b binding protein 1 (Lhcb1). This demonstrated that OsCpn60α1 is required for the folding of rbcL and that failure of that process is seedling-lethal.     

The search for the missing link: a relic plastid in Perkinsus?

Perkinsus marinus (Phylum Perkinsozoa) is a protozoan parasite that has devastated natural and farmed oyster populations in the USA, significantly affecting the shellfish industry and the estuarine environment. The other two genera in the phylum, Parvilucifera and Rastrimonas, are parasites of microeukaryotes. The Perkinsozoa occupies a key position at the base of the dinoflagellate branch, close to its divergence from the Apicomplexa, a clade that includes parasitic protista, many harbouring a relic plastid. Thus, as a taxon that has also evolved toward parasitism, the Perkinsozoa has attracted the attention of biologists interested in the evolution of this organelle, both in its ultrastructure and the conservation, loss or transfer of its genes. A review of the recent literature reveals mounting evidence in support of the presence of a relic plastid in P. marinus, including the presence of multimembrane structures, characteristic metabolic pathways and proteins with a bipartite N-terminal extension. Further, these findings raise intriguing questions regarding the potential functions and unique adaptation of the putative plastid and/or plastid genes in the Perkinsozoa. In this review we analyse the above-mentioned evidence and evaluate the potential future directions and expected benefits of addressing such questions. Given the rapidly expanding molecular/genetic resources and methodological toolbox for Perkinsus spp., these organisms should complement the currently established models for investigating plastid evolution within the Chromalveolata.     

Characterization of the 16S–23S internal transcribed spacer among 34 higher plants: suitability for interspecific plastid transformation

Biomanufacturing by chloroplast transgene expression has the potential to produce significant amounts of biopharmaceuticals, endow plants with novel commercial or humanitarian capabilities, enhance phytoremediation methods and harden plants against adverse environments. Plastid bioengineering exploits the phenomenon of homologous recombination to specifically integrate heterologous sequences into the plastid genome. Previous research suggests the plastid genome 16S–23S internal transcribed spacer provides an advantageous integration site for transgene expression. To characterize the suitability of the 16S–23S region for interspecific recombination, we developed primers against conserved plastid sequences and amplified ∼2.6 kb from 25 plant species. We analyzed the amplicons with nine species from Genbank for homeology, phylogenetic relationships, potential to form chimeric rDNA elements disruptive to translational/replication systems, and the potential number of recombination events for various minimal essential processing segments (MEPS) lengths. Multiple sequence alignment of the 34 species revealed considerable conservation, with identities exceeding 95% among the angiosperms. Substitutions were statistically clustered, generally in noncoding sites, although proposed functional elements such as the OriA region and 3′ terminus of the 16S rRNA exhibited unexpected variation. The nonrandom distribution of substitutions undermines the established, statistical method of estimating the number of recombination initiation sites. This finding is further substantiated by comparing statistical estimates of the number of MEPS sites to a direct count at three different MEPS lengths. We frame this in silico analysis in terms of the potential of the 16S–23S region as a target for interspecific transformation, and describe a ‘primer-to-plastid’ system to rapidly generate species-specific flanking regions for transformation vectors.     

Free‐running circadian rhythm of plastid movements in individual cells of pyrocystis lunula (dinophyta)

Abstract

Pyrocystis lunula, a dinoflagellate lacking periodicity in spontaneous bio‐luminescence, shows a characteristic circadian rhythm of plastid movements which can be monitored photographically in individual cells. The rhythm persists under free‐running conditions in constant dim light (40–50 Ix), but is damped out already in LL of 100 lx.     

Occurrence of unidentified bodies that resemble “plastid initials” in poplar cells after breaking of dormancy in midwinter

Summary Electron-microscopic studies of plastids in cortical cells of poplar (Populus euramericana cv. gelrica) were carried out to examine whether any structural changes were initiated after breaking of dormancy in midwinter under non-growing conditions. After the breaking of dormancy, ultrastructural changes became evident and the profiles of plastids became heterogeneous. Organelles resembling the plastid initials proposed by Mühlenthaler and Frey-Wyssling in 1965 were frequently observed concomitant with changes in the plastid envelope. The formation of plastid initials appeared to be initiated by the formation of septa in pre-existing plastids. After this stage, narrow connections appeared between the initials and the parent plastids. Approximately 50 days after the breaking of dormancy in late March, further heterogeneity in the profiles of plastids was observed. At this stage, young plastids (plastids without starch granules) were frequently observed and the formation of plastid initials was hardly ever observed. These observations suggest that the plastid initials may be present for only a limited period in the cortical cells of the poplar and may be the precursors of the proplastids. Similar ultrastructural profiles were found in cortical cells of mulberry and in leaf buds of apple trees, suggesting that such changes in the ultrastructure of plastids are a general feature of perennials.     

Analysis of plastid and nuclear DNA data in plant phylogenetics——evaluation and improvement

Correct combination of plastid(cp)and nuclear(nr)DNA data for plant phylogenetic reconstructions is not a new issue,but with an increasing number of nrDNA loci being used,it is of ever greater practical concern.For accurately reconstructing the phylogeny and evolutionary history of plant groups,correct treatment of phylogenetic incongruence is a vital step in the proper analysis of cpDNA and nrDNA data.We first evaluated the current status of analyzing cpDNA and nrDNA data by searching all articles published in the journal Systematic Botany between 2005 and 2011.Many studies combining cpDNA and nrDNA data did not rigorously assess the combinability of the data sets,or did not address in detail possible reasons for incongruence between the two data sets.By reviewing various methods,we outline a procedure to more accurately analyze and/or combine cpDNA and nrDNA data,which includes four steps:identifying significant incongruence,determining conflicting taxa,providing possible interpretations for incongruence,and reconstructing the phylogeny after treating incongruence.Particular attention is given to explanation of the cause of incongruence.We hope that our procedure will help raise awareness of the importance of rigorous analysis and help identify the cause of incongruence before combining cpDNA and nrDNA data.     

Effect of cortisone on the developmental pattern of the neutral and the acid β-galactosidases of the small intestine of the rat

1. The developmental pattern and effect of cortisone on acid beta-galactosidase and neutral beta-galactosidase were studied in postnatal rats by a recently proposed method for their independent determination. 2. After birth the acid beta-galactosidase activity increases in the ileum, whereas it decreases slightly in the jejunum. On day 16 after birth the activity in the ileum decreases and in 20-day-old rats activity in both parts of the intestine decreases to adult values. In suckling animals the activity in the ileum exceeds the jejunal activity severalfold and in adult animals the activity in the jejunum is slightly higher than that in the ileum. 3. Neutral beta-galactosidase activity is high after birth and decreases in both jejunum and ileum after day 20 after birth. In 12-20-day-old rats activity in both parts is essentially the same, but in adult animals jejunal activity exceeds ileal activity four-to five-fold. 4. Cortisone (0.5, 2.0 or 5.0mg/100g body wt. daily for 4 days) does not influence the activity of either enzyme in 60-day-old rats. Acid beta-galactosidase activity is decreased after cortisone treatment in 8-, 12-, 16-and 18-day-old rats, with sensitivity to cortisone increasing with the approach of weaning. No effect of cortisone on acid beta-galactosidase is seen in 8-day-old rats. Neutral beta-galactosidase activity is increased in the ileum of 8-, 12-, 16- and 18-day old rats, but only in the jejunum of 8-and 12-day-old rats.