Molecular, functional and ultrastructural characterisation of plastids from six species of the parasitic flowering plant genus Cuscuta.

Plastids of Cuscuta reflexa Roxb., C. subinclusa D. et H., C. gronovii Willd. and C. campestris Yunck. possess thylakoids and contain both chlorophyll a and b in a ratio similar to that of stem tissue of the systematically closely related but 'normal' green Ipomoea tricolor. In contrast, plastids of C. odorata R. et P. and C. grandiflora H.B.K. do not contain any chlorophyll or possess thylakoids. Light-driven electron transport, as measured by oxygen evolution and indicated by analysis of chlorophyll fluorescence, was present in all chlorophyll-containing species. The photosystem II efficiency was low and ranged from 0.511 to 0.687. The plastid rbcL gene could not be detected in C. odorata, but was present in all other tested species. Neither rbcL transcripts nor the large subunit of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) could be detected in C. odorata and C. grandiflora. Low amounts of the large subunit of Rubisco were detected immunologically in all other Cuscuta species. Apparently, the genus Cuscuta comprises species with different degrees of plastid functionality, ranging from intact chloroplasts, via plastids with impaired protein production and gene expression to plastids with reduced plastome gene content.

The implication of a plastid-derived factor in the transcriptional control of nuclear genes encoding the light-harvesting chlorophyll a/b protein.

In carotenoid-deficient albina mutants of barley and in barley plants treated with the herbicide Norflurazon the light-dependent accumulation of the mRNA for the light-harvesting chlorophyll a/b protein (LHCP) is blocked. Thus, the elimination of a functional chloroplast, either as a result of mutation or as a result of herbicide treatment, can lead to the specific suppression of the expression of a nuclear gene encoding a plastid-localized protein. These results confirm and extend earlier observations on maize [Mayfield and Taylor (1984) Eur. J. Biochem. 144, 79-84]. The inhibition of mRNA accumulation appears to be specific for the LHCP; the mRNAs encoding the small subunit of ribulose-1,5-bisphosphate carboxylase and the NADPH: protochlorophyllide oxidoreductase are relatively unaffected. The failure of the albina mutants and of Norflurazon-treated plants to accumulate the LHCP mRNA is not exclusively caused by an instability of the transcript but rather by the inability of the plants to enhance the rate of transcription of the LHCP genes during illumination. Several chlorophyll-deficient xantha mutants of barley, which are blocked after protoporphyrin IX or Mg-protoporphyrin, and the chlorophyll-b-less mutant chlorina f2 accumulate the LHCP mRNA to almost normal levels during illumination. Thus, if any of the reactions leading to chlorophyll formation is involved in the control of LHCP mRNA accumulation it should be one between the formation of protochlorophyllide and the esterification of chlorophyllide a. While the nature of the regulatory factor(s) has not been identified our results suggest that, in addition to phytochrome (Pfr), plastid-dependent factors are required for a continuous light-dependent transcription of nuclear genes encoding the LHCP.

Filament formation in vitro of a sieve tube protein from Cucurbita maxima and Cucurbita pepo.

Phloem proteins of the sieve tube exudate from Cucurbita maxima Duch. and Cucurbita pepo L. were investigated as to their filament forming ability in vitro. From the two main proteins (116000 dalton, 30000 dalton) only the 116000 dalton protein was found to form reversibly distinct filaments of 6-7 nm diameter upon removal of SH-protecting agents from the buffer, whereas the 30000 dalton protein was precipitated as amorphous material under these conditions. The protein filaments were similar to the filaments ocurring within the sieve tube cells in vivo.

Protein filaments-structural components of the phloem exudate : I. Observations with Cucurbita and Nicotiana.

Fine structure and chemical composition of the phloem exudate of Cucurbita maxima and Nicotaiana glauca x suaveolens are investigated. Filamentous structures, several microns in length, are identified as structural components of the exudate by means of negative staining and electron microscopy. Two types of filaments are described: one form measures nearly 40 Å in diameter and shows a beaded appearance with regular spacings of about 50 Å; it is termed "elementary filament". The second form has a diameter of about 90 Å and presumably consists of two helically arranged 40 Å subunits.The proteinaceous nature of the filaments is indicated by chemical analysis. The main macromolecular component of the exudate is demonstrated to be protein. Only traces of nucleic acids are detectable, lipids and polysaccharides cannot be found. The identity of the protein filaments with the filamentous structures ("slime", "P-protein"), as revealed in thin sections of mature sieve tubes, is discussed.

Fine structure of mycorrhiza in Neottia nidus-avis (L.) L. C. Rich. (Orchidaceae).

The fine structural features of the symbiotic relationship between Neottia and the fungus Rhizoctonia have been examined. Different stages of development of the fungus within the orchid's root cells are described. The fungus-attacked Neottia cells show striking changes. The central vacuole is partly filled by a conspicuous plasmatic network while the nucleus enlarges considerably. Of special interest is the development of an extended rough-surfaced ER.

[On the plasmatic filaments in assimilate conducting cells, their development and fine structure]. / Zur Herkunft und Struktur der Plasmafilamente in Assimilatleitbahnen.

Taking into account the literature on the so-called sieve-tube slime ("mictoplasm", "slime strands") and regarding its fine structure more in detail the term plasmatic filament ("Plasmafilament") is proposed and will be used in this paper to characterize the individual exceedingly fine subunit of the plasmatic network (or slime) in sieve elements. Up to now plasmatic filaments have mostly been erroneously called "fibrils". The dimension of a fibrill has now been defined anew and differentiated from its subunit "plasmatic filament".In the first part of these investigations some aspects of the development of plasmatic filaments and their spreading over the total lumen of Dioscorea sieve elements will be reported.Previous to the first appearance of filaments the later sieve element abounds in plasmatic components, the groundplasm being extremely rich in ribosomes (Fig. 1). The difference between young sieve elements and the neighbouring parenchyma cells is nearly imperceptible apart from a slight variation in ribosome density. Plastids are very useful in distinguishing these two cell types from each other. The development of osmiophilic inclusions that characterize sieve-element plastids in Dioscorea has already been initiated in these very young cells.The earliest stages in the formation of plasmatic filaments that up to now have been revealed in Dioscorea show masses of filaments, some short and granular in appearance (Fig. 2: *), some already elngated and filamentous (Fig. 2: F). After expanding over the entire cell those filaments still look like having their origin directly in groundplasm (Fig. 5). Elements of the ER-system and many ribosomes cross the plasmatic filaments during all developmental stages of their network, which is at no time surrounded by any membrane.In sieve elements of Dioscorea, Primula, Cuscuta and Cucumis our investigations furthermore yielded some detail on the filament substructure. A cross-sectioned plasmatic filament is composed of an osmiophilic outer ring with a light centre (Fig. 11) corresponding in a longitudinal view to two deeply contrasted outer layers and an inner one without any contrast (Fig. 8). An individual filament has an overall diameter of 120-150 Å and an up to now indeterminable length that exceeds at least several microns.The real nature of these fine structures will be discussed in relation to similar structures and their meaning in plant and animal cells.