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Stefan A Rensing - One of the best experts on this subject based on the ideXlab platform.
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a single target mitochondrial rna editing factor of Funaria hygrometrica can fully reconstitute rna editing at two sites in physcomitrella patens
Plant and Cell Physiology, 2017Co-Authors: Mareike Schallenbergrudinger, Bastian Oldenkott, Manuel Hiss, Phuong Le Trinh, Volker Knoop, Stefan A RensingAbstract:Nuclear-encoded pentatricopeptide repeat (PPR) proteins are key factors for site-specific RNA editing, converting cytidines into uridines in plant mitochondria and chloroplasts. All editing factors in the model moss Physcomitrella patens have a C-terminal DYW domain with similarity to cytidine deaminase. However, numerous editing factors in flowering plants lack such a terminal DYW domain, questioning its immediate role in the pyrimidine base conversion process. Here we further investigate the Physcomitrella DYW-type PPR protein PPR_78, responsible for mitochondrial editing sites cox1eU755SL and rps14eU137SL. Complementation assays with truncated proteins demonstrate that the DYW domain is essential for full PPR_78 editing functionality. The DYW domain can be replaced, however, with its counterpart from another editing factor, PPR_79. The PPR_78 ortholog of the related moss Funaria hygrometrica fully complements the Physcomitrella mutant for editing at both sites, although the editing site in rps14 is lacking in Funaria. Editing factor orthologs in different taxa may thus retain editing capacity for multiple sites despite the absence of editing requirement.
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assigning dyw type ppr proteins to rna editing sites in the funariid mosses physcomitrella patens and Funaria hygrometrica
Plant Journal, 2011Co-Authors: Mareike Rudinger, Stefan A Rensing, Peter Szovenyi, Volker KnoopAbstract:The plant-specific pentatricopeptide repeat (PPR) proteins with variable PPR repeat lengths (PLS-type) and protein extensions up to the carboxyterminal DYW domain have received attention as specific recognition factors for the C-to-U type of RNA editing events in plant organelles. Here, we report a DYW-protein knockout in the model plant Physcomitrella patens specifically affecting mitochondrial RNA editing positions cox1eU755SL and rps14eU137SL. Assignment of DYW proteins and RNA editing sites might best be corroborated by data from a taxon with a slightly different, yet similarly manageable low number of editing sites and DYW proteins. To this end we investigated the mitochondrial editing status of the related funariid moss Funaria hygrometrica. We find that: (i) Funaria lacks three mitochondrial RNA editing positions present in Physcomitrella, (ii) that F. hygrometrica cDNA sequence data identify nine DYW proteins as clear orthologues of their P. patens counterparts, and (iii) that the 'missing' 10th DYW protein in F. hygrometrica is responsible for two mitochondrial editing sites in P. patens lacking in F. hygrometrica (nad3eU230SL, nad4eU272SL). Interestingly, the third site of RNA editing missing in F. hygrometrica (rps14eU137SL) is addressed by the DYW protein characterized here and the presence of its orthologue in F. hygrometrica is explained through its simultaneous action on site cox1eU755SL conserved in both mosses.
Jessica M Budke - One of the best experts on this subject based on the ideXlab platform.
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the moss Funaria hygrometrica has cuticular wax similar to vascular plants with distinct composition on leafy gametophyte calyptra and sporophyte capsule surfaces
Annals of Botany, 2016Co-Authors: Lucas Busta, Jessica M Budke, Reinhard JetterAbstract:BACKGROUND AND AIMS Aerial surfaces of land plants are covered with a waxy cuticle to protect against water loss. The amount and composition of cuticular waxes on moss surfaces had rarely been investigated. Accordingly, the degree of similarity between moss and vascular plant waxes, and between maternal and offspring moss structure waxes is unknown. To resolve these issues, this study aimed at providing a comprehensive analysis of the waxes on the leafy gametophyte, gametophyte calyptra and sporophyte capsule of the moss Funaria hygrometrica METHODS Waxes were extracted from the surfaces of leafy gametophytes, gametophyte calyptrae and sporophyte capsules, separated by gas chromatography, identified qualitatively with mass spectrometry, and quantified with flame ionization detection. Diagnostic mass spectral peaks were used to determine the isomer composition of wax esters. KEY RESULTS The surfaces of the leafy gametophyte, calyptra and sporophyte capsule of F. hygrometrica were covered with 0·94, 2·0 and 0·44 μg cm(-2) wax, respectively. While each wax mixture was composed of mainly fatty acid alkyl esters, the waxes from maternal and offspring structures had unique compositional markers. β-Hydroxy fatty acid alkyl esters were limited to the leafy gametophyte and calyptra, while alkanes, aldehydes and diol esters were restricted to the sporophyte capsule. Ubiquitous fatty acids, alcohols, fatty acid alkyl esters, aldehydes and alkanes were all found on at least one surface. CONCLUSIONS This is the first study to determine wax coverage (μg cm(-2)) on a moss surface, enabling direct comparisons with vascular plants, which were shown to have an equal amount or more wax than F. hygrometrica Wax ester biosynthesis is of particular importance in this species, and the ester-forming enzyme(s) in different parts of the moss may have different substrate preferences. Furthermore, the alkane-forming wax biosynthesis pathway, found widely in vascular plants, is active in the sporophyte capsule, but not in the leafy gametophyte or calyptra. Overall, wax composition and coverage on F. hygrometrica were similar to those reported for some vascular plant species, suggesting that the underlying biosynthetic processes in plants of both lineages were inherited from a common ancestor.
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identification of β hydroxy fatty acid esters and primary secondary alkanediol esters in cuticular waxes of the moss Funaria hygrometrica
Phytochemistry, 2016Co-Authors: Lucas Busta, Jessica M Budke, Reinhard JetterAbstract:The plant cuticle, a multi-layered membrane that covers plant aerial surfaces to prevent desiccation, consists of the structural polymer cutin and surface-sealing waxes. Cuticular waxes are complex mixtures of ubiquitous, typically monofunctional fatty acid derivatives and taxon-specific, frequently bifunctional specialty compounds. To further our understanding of the chemical diversity of specialty compounds, the waxes on the aerial structures of the leafy gametophyte, sporophyte capsule, and calyptra of the moss Funaria hygrometrica were surveyed. Respective moss surfaces were extracted, and resulting lipid mixtures were analyzed by gas chromatography-mass spectrometry (GC-MS). The extracts contained ubiquitous wax compound classes along with two prominent, unidentified classes of compounds that exhibited some characteristics of bifunctional structures. Microscale transformations led to derivatives with characteristic MS fragmentation patterns suggesting possible structures for these compounds. To confirm the tentative structure assignments, one compound in each of the suspected homologous series was synthesized. Based on GC-MS comparison with the authentic standards, the first series of compounds was identified as containing esters formed by β-hydroxy fatty acids and wax alcohols, with ester chain lengths varying from C42 to C50 and the most prominent homolog being C46. The second series consisted of fatty acid esters of 1,7-alkanediols, linked via the primary hydroxyl group, with ester chain lengths C40-C52 also dominated by the C46 homolog. The β-hydroxy acid esters were restricted to the sporophyte capsule, and the diol esters to the leafy gametophyte and calyptra. Based on their homolog and isomer distributions, and the presence of free 1,7-triacontanediol, possible biosynthetic reactions leading to these compounds are discussed.
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dehydration protection provided by a maternal cuticle improves offspring fitness in the moss Funaria hygrometrica
Annals of Botany, 2013Co-Authors: Jessica M Budke, Bernard Goffinet, Cynthia S. JonesAbstract:†Background and Aims In bryophytes the sporophyte offspring are in contact with, nourished from, and partially surrounded by the maternal gametophyte throughout their lifespan. During early development, the moss sporophyte is covered by the calyptra, a cap of maternal gametophyte tissue that has a multilayered cuticle. In this study the effects on sporophyte offspring fitness of removing the maternal calyptra cuticle, in combination with dehydration stress, is experimentally determined. †Methods Using the moss Funaria hygrometrica, calyptra cuticle waxes were removed by chemical extraction and individuals were exposed to a short-term dehydration event. Sporophytes were returned to high humidity to complete development and then aspects of sporophyte survival, development, functional morphology, and reproductive output were measured. †Key Results It was found that removal of calyptra cuticle under low humidity results in significant negative impacts to moss sporophyte fitness, resulting in decreased survival, increased tissue damage, incomplete sporophyte development, more peristome malformations, and decreased reproductive output. †Conclusions This study represents the strongest evidence to date that the structure of the calyptra cuticle functions in dehydration protection of the immature moss sporophyte. The investment in a maternal calyptra with a multilayered cuticle increases offspring fitness and provides a functional explanation for calyptra retention across mosses. The moss calyptra may represent the earliest occurance of maternal protection via structural provisioning of a cuticle in green plants.
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examining the ability of calyptrae to produce protonema in Funaria hygrometrica
Evansia, 2012Co-Authors: Melissa A Wynne, Jessica M BudkeAbstract:Abstract. Our research assesses the ability of calyptrae to produce protonema in the moss species Funaria hygrometrica Hedw. Herein the following question is addressed: how long after detachment from the maternal plant are calyptrae able to produce new individuals by means of protonema? Plants from a local Connecticut population were grown in laboratory conditions until 2, 14, or 28 days post-detachment of the calyptra from the leafy gametophyte. Calyptrae were removed, placed onto sterile media, and observed for protonemal growth for 10 weeks. Calyptrae were found to be alive and produced protonema at all three developmental ages. The youngest calyptrae produced the highest percentage of protonemal growth, whereas the oldest calyptrae produced significantly fewer. Our data provide evidence that calyptrae in a laboratory setting remain alive after detachment from the maternal plant and may have the potential to act as a dispersal unit.
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The cuticle on the gametophyte calyptra matures before the sporophyte cuticle in the moss Funaria hygrometrica (Funariaceae).
American Journal of Botany, 2012Co-Authors: Jessica M Budke, Bernard Goffinet, Cynthia S. JonesAbstract: Premise of the study: In vascular plants, leaf primordia prevent desiccation of the shoot apical meristem. Lacking leaves, the undifferentiated moss sporophyte apex is covered by the calyptra, a cap of maternal gametophyte tissue that is hypothesized to function in desiccation protection. Herein, we compare cuticle development on the calyptra and sporophyte to assess the calyptra ’ s potential to protect the sporophyte from desiccation. As the fi rst comprehensive study of moss sporophyte cuticle development, this research broadens our perspectives on cuticle development and evolution across embryophytes. Methods: Calyptrae and sporophytes at nine developmental stages were collected from a laboratory-grown population of the moss Funaria hygrometrica . Tissues were embedded, sectioned, then examined using transmission electron microscopy. Epidermal cells were measured for thickness of the cuticle layers, cell wall thickness, and lumen size. Key results: The calyptra cuticle develops precociously and reaches maturity before the sporophyte cuticle. Calyptrae are covered by a four-layered cuticle at all stages, whereas sporophyte cuticle maturation is delayed until sporangium formation. The development and thickening of the sporophyte cuticle occurs in an acropetal wave. Conclusions: A multilayered calyptra cuticle at the earliest developmental stages is consistent with its ability to protect the immature sporophyte from desiccation. Young sporophytes lack a complex cuticle and thus may require protection, whereas in older sporophytes a mature cuticle develops. The moss calyptra is not a vestigial structure, but rather the calyptra ’ s role in preventing desiccation offers a functional explanation for calyptra retention during the 450 Myr of moss evolution.
Reinhard Jetter - One of the best experts on this subject based on the ideXlab platform.
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the moss Funaria hygrometrica has cuticular wax similar to vascular plants with distinct composition on leafy gametophyte calyptra and sporophyte capsule surfaces
Annals of Botany, 2016Co-Authors: Lucas Busta, Jessica M Budke, Reinhard JetterAbstract:BACKGROUND AND AIMS Aerial surfaces of land plants are covered with a waxy cuticle to protect against water loss. The amount and composition of cuticular waxes on moss surfaces had rarely been investigated. Accordingly, the degree of similarity between moss and vascular plant waxes, and between maternal and offspring moss structure waxes is unknown. To resolve these issues, this study aimed at providing a comprehensive analysis of the waxes on the leafy gametophyte, gametophyte calyptra and sporophyte capsule of the moss Funaria hygrometrica METHODS Waxes were extracted from the surfaces of leafy gametophytes, gametophyte calyptrae and sporophyte capsules, separated by gas chromatography, identified qualitatively with mass spectrometry, and quantified with flame ionization detection. Diagnostic mass spectral peaks were used to determine the isomer composition of wax esters. KEY RESULTS The surfaces of the leafy gametophyte, calyptra and sporophyte capsule of F. hygrometrica were covered with 0·94, 2·0 and 0·44 μg cm(-2) wax, respectively. While each wax mixture was composed of mainly fatty acid alkyl esters, the waxes from maternal and offspring structures had unique compositional markers. β-Hydroxy fatty acid alkyl esters were limited to the leafy gametophyte and calyptra, while alkanes, aldehydes and diol esters were restricted to the sporophyte capsule. Ubiquitous fatty acids, alcohols, fatty acid alkyl esters, aldehydes and alkanes were all found on at least one surface. CONCLUSIONS This is the first study to determine wax coverage (μg cm(-2)) on a moss surface, enabling direct comparisons with vascular plants, which were shown to have an equal amount or more wax than F. hygrometrica Wax ester biosynthesis is of particular importance in this species, and the ester-forming enzyme(s) in different parts of the moss may have different substrate preferences. Furthermore, the alkane-forming wax biosynthesis pathway, found widely in vascular plants, is active in the sporophyte capsule, but not in the leafy gametophyte or calyptra. Overall, wax composition and coverage on F. hygrometrica were similar to those reported for some vascular plant species, suggesting that the underlying biosynthetic processes in plants of both lineages were inherited from a common ancestor.
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identification of β hydroxy fatty acid esters and primary secondary alkanediol esters in cuticular waxes of the moss Funaria hygrometrica
Phytochemistry, 2016Co-Authors: Lucas Busta, Jessica M Budke, Reinhard JetterAbstract:The plant cuticle, a multi-layered membrane that covers plant aerial surfaces to prevent desiccation, consists of the structural polymer cutin and surface-sealing waxes. Cuticular waxes are complex mixtures of ubiquitous, typically monofunctional fatty acid derivatives and taxon-specific, frequently bifunctional specialty compounds. To further our understanding of the chemical diversity of specialty compounds, the waxes on the aerial structures of the leafy gametophyte, sporophyte capsule, and calyptra of the moss Funaria hygrometrica were surveyed. Respective moss surfaces were extracted, and resulting lipid mixtures were analyzed by gas chromatography-mass spectrometry (GC-MS). The extracts contained ubiquitous wax compound classes along with two prominent, unidentified classes of compounds that exhibited some characteristics of bifunctional structures. Microscale transformations led to derivatives with characteristic MS fragmentation patterns suggesting possible structures for these compounds. To confirm the tentative structure assignments, one compound in each of the suspected homologous series was synthesized. Based on GC-MS comparison with the authentic standards, the first series of compounds was identified as containing esters formed by β-hydroxy fatty acids and wax alcohols, with ester chain lengths varying from C42 to C50 and the most prominent homolog being C46. The second series consisted of fatty acid esters of 1,7-alkanediols, linked via the primary hydroxyl group, with ester chain lengths C40-C52 also dominated by the C46 homolog. The β-hydroxy acid esters were restricted to the sporophyte capsule, and the diol esters to the leafy gametophyte and calyptra. Based on their homolog and isomer distributions, and the presence of free 1,7-triacontanediol, possible biosynthetic reactions leading to these compounds are discussed.
Mareike Schallenbergrudinger - One of the best experts on this subject based on the ideXlab platform.
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a single target mitochondrial rna editing factor of Funaria hygrometrica can fully reconstitute rna editing at two sites in physcomitrella patens
Plant and Cell Physiology, 2017Co-Authors: Mareike Schallenbergrudinger, Bastian Oldenkott, Manuel Hiss, Phuong Le Trinh, Volker Knoop, Stefan A RensingAbstract:Nuclear-encoded pentatricopeptide repeat (PPR) proteins are key factors for site-specific RNA editing, converting cytidines into uridines in plant mitochondria and chloroplasts. All editing factors in the model moss Physcomitrella patens have a C-terminal DYW domain with similarity to cytidine deaminase. However, numerous editing factors in flowering plants lack such a terminal DYW domain, questioning its immediate role in the pyrimidine base conversion process. Here we further investigate the Physcomitrella DYW-type PPR protein PPR_78, responsible for mitochondrial editing sites cox1eU755SL and rps14eU137SL. Complementation assays with truncated proteins demonstrate that the DYW domain is essential for full PPR_78 editing functionality. The DYW domain can be replaced, however, with its counterpart from another editing factor, PPR_79. The PPR_78 ortholog of the related moss Funaria hygrometrica fully complements the Physcomitrella mutant for editing at both sites, although the editing site in rps14 is lacking in Funaria. Editing factor orthologs in different taxa may thus retain editing capacity for multiple sites despite the absence of editing requirement.
Jeffrey G Duckett - One of the best experts on this subject based on the ideXlab platform.
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nuclear differentiation in the tilamentous caulonema of the moss Funaria hygrometrica
2016Co-Authors: Keith I Kingham, Jeffrey G Duckett, Matthew C P Glyn, Andrew R LeitchAbstract:Nuclei from different cell types in plants and animals show many features of differentiation; they differ in shape, volume, structure, ultrastructure and in the distribution of nuclear components. Using the filamentous caulonema of the moss Funaria hygrometrica Hedw. this study records the changes in cytoplasmic organization alongside the reorganization of the interphase nucleus. Events taking place in the meristematic cells at or near the tip of the advancing caulonemal filaments (e.g. acquisition of polarity, tip growth, nuclear and cell division, side branch initiation) are associated with haploid nuclei (1C DNA amount 0 5 pg) that are spherical or slightly oval, with no blocks of condensed chromatin, and a large central nucleolus with a large granular component. Maturation of the caulonemal cells involves wall thickening and pigmentation concomitant with suspension of elongate plastids in linear arrays along endoplasmic strands. Many cells become highly polarized with the majority of the organelles at their apical ends. These cytoplasmic changes are associated with endoreduplication of the genome to about 8C. Endoreduplication occurs by amplification of the 1 C genome to give nuclei with 1C-8C DNA amounts. There is no evidence of differential amplification of the genome. The amplification in the copy number of ribosomal RNA genes is associated with the heterochromatinisation of the genes within the nucleolus. At the same time the nucleolus reduces in volume owing to a diminution of the granular component and all components of the nucleolus become spatially separate. There is an increased nuclear volume associated with endoreduplication and the nucleus elongates causing an increase in the surface area of the nuclear envelope. These major nuclear reorganizations are associated with a stable distribution of the 'D' polypeptide involved in pre-mRNA splicing. Scrutiny of published data suggests that similar differentiation events might be encountered commonly in other organisms. The changing nuclear morphology probably reflects the changing activity of the nucleus and the cell. It might be that nuclear reorganization changes the balance of genes or gene products and the spatial distribution of the component parts to enable the new nuclear functions. These results suggest that nuclear differentiation is a fundamental feature of cell differentiation.
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ephemerella readeri mull hal physcomitrella readeri mull hal i g stone g a m scott funariidae bryophyta a genus and species new to europe
Journal of Bryology, 2010Co-Authors: E J Hooper, Jeffrey G Duckett, A C Cuming, W E Kunin, Silvia PresselAbstract:AbstractA morphological and molecular analysis of a Physcomitrella, collected from a reservoir margin in the north of England, revealed this to be P. readeri, a species new to Europe. The present study clarifies previous confusion over the taxonomy of P. readeri showing it to be clearly distinct in both sporophytic and gametophytic characters from P. patens and uniform across its world range from England to USA, Japan and Australasia. While phylograms of the ITS1 region from both the Physcomitrella species, Physcomitrium pyriforme (Hedw.) Bruch & Schimp., Enthosthodon attenuatus (Dicks.) Bryhn and Funaria hygrometrica Hedw., place the first two in separate clades, in ITS2 phylograms they occur as sister taxa. This, together with previous genealogical studies on the speciation history of the Physcomitrella–Physcomitrium species complex, and morphology, suggests that generic rank is appropriate for P. readeri. We therefore reinstate the original name Ephemerella readeri Mull. Hal. Recent records at several ...
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the role of dna methylation on nuclear and cell differentiation in the filamentous caulonema of the moss Funaria hygrometrica
New Phytologist, 1998Co-Authors: Keith I Kingham, Jeffrey G Duckett, Blanka Gazdova, Ales Kovarik, Milan Bezdek, Andrew R LeitchAbstract:SUMMARY This paper describes the effects of hypomethylation with 5-azacytidine (5azaC) and dihydroxypropyladenine (DHPA) on protonemata of the moss Funaria hygrometrica Hedw. Following treatment with 5azaC or DHPA, hypomethylation of the EcoR II (CCA(T)GG) and Pst I (CTGCAG) sites in ribosomal DNA (rDNA) was confirmed using restriction enzyme analysis and Southern hybridization. Hypomethylation of the genome had profound effects on protonemal differentiation. Whilst apical cell organization and cell dimensions and shape remained unchanged, there was a marked retardation in both cytoplasmic and nuclear differentiation. Developmental abnormalities included: late and erratic side branch formation, some loss of the distinction between chloronema and caulonema, formation of aberrant buds, and loss of the potential to form brood and tmema cells after 5azaC treatment which in general had more profound effects than DHPA. Cytologically, caulonema cells were less highly polarized or unpolarized, and tended to retain small round chloroplasts, whilst nuclei in the hypomethylated protonema endoreduplicated to lower levels and tended to remain more spherical. Increased nucleolar volumes and loss of intranucleolar rDNA heterochromatin following hypomethylation might be the result of increased transcriptional activity of ribosomal RNA (rRNA) genes and drug-induced DNA decondensation, respectively. Growing hypomethylated protonema at 25 ?C induced extremely atypical cells and development. This temperature sensitivity and aberrations in development overall can be attributed to changes in the normal patterns of gene expression brought about by hypomethylation of gene promoter or regulator regions.
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nuclear differentiation in the filamentous caulonema of the moss Funaria hygrometrica
New Phytologist, 1995Co-Authors: Keith I Kingham, Jeffrey G Duckett, Matthew C P Glyn, Andrew R LeitchAbstract:Nuclei from different cell types in plants and animals show many features of differentiation; they differ in shape, volume, structure, ultrastructure and in the distribution of nuclear components. Using the filamentous caulonema of the moss Funaria hygrometrica Hedw. this study records the changes in cytoplasmic organization alongside the reorganization of the interphase nucleus, Events taking place in the meristematic cells at or near the lip of the advancing caulonemal filaments (e.g. acquisition of polarity, tip growth, nuclear and cell division, side branch initiation] are associated with haploid nuclei (1C DXA amount 0.5 pg) that are spherical or slightly oval, with no blocks of condensed chromatin, and a large central nucleolus with a large granular component. Maturation of the caulonemal cells involves wall thickening and pigmentation concomitant with suspension of elongate plastids in linear arrays along endoplasmic strands. Many cells become highly polarized with the majority of the organelles at their apical ends. These eytoplasmic changes are associated with endoreduplication of the genome to about 8C, endoreduplication occurs by amplification of the 1C genome to give nuclei with IC-SC DNA amounts. There is no evidence of differential amplification of the genome. The amplification in the copy number of ribosomal RNA genes is associated with the heterochromatinisation of the genes within the nucleolus. At the same time the nucleolus reduces in volume owing to a diminution of the granular component and all components of the nucleolus become spatial separate. There is an increased nuclear volume associated with endoreduplication and the nucleus elongates causing an increase in the surface area of the nuclear envelope. These major nuclear reorganizations are associated with a stable distribution of the 'D' polypeptide involved in pre-mRNA splicing. Scrutiny of published data suggests that similar differentiation events might be encountered commonly in other organisms. The changing nuclear morphology probably reflects the changing activity of the nucleus and the cell. It might be that nuclear reorganization changes the balance of genes or gene products and the spatial distribution of the component pans to enable the new nuclear functions. These results suggest that nuclear differentiation is a fundamental feature of cell differentiation.
