Pinus silvestris and Pinus halepensis trees grown in Germany and Spain, respectively, showing abnormal shoot branching, dwarfed needles and other symptoms were examined for the presence of plant-pathogenic mollicutes (phytoplasmas). While phytoplasmas could not be detected unambiguously with microscopical methods, PCR amplification using universal phytoplasma primers yielded positive results. Samples collected from symptomatic and non-symptomatic plant parts of both symptomatic Pinus silvestris and Pinus halepensis trees tested positive. Also, surrounding non-symptomatic trees proved to be phytoplasma-infected. Comparisons revealed that the 16S rRNA gene sequences of the phytoplasmas identified in Pinus silvestris and Pinus halepensis were nearly identical. However, the pine phytoplasma is only distantly related to other phytoplasmas. The closest relatives are members of the palm lethal yellowing and rice yellow dwarf groups and ‘Candidatus Phytoplasma castaneae’, which share between 94·5 and 96·6 % 16S rRNA gene sequence similarity. From these data it can be concluded that the phytoplasmas identified in the two Pinus species represent a coherent but discrete taxon; it is proposed that this taxon be distinguished at putative species level under the name ‘Candidatus Phytoplasma pini’.

Nondispersive protein bodies present in the sieve elements in addition to dispersive P-protein are characteristic features of many woody dicotyledons; their origin may be nuclear or cytoplasmic. While nuclear nondispersive protein bodies are found in only two families, the Boraginaceae and Myristicaceae, bodies of cytoplasmic origin are present in 39 of the more than 350 families screened. These results were obtained from 228 dicotyledons studied with the electron microscope and data of additional species from the literature. The terminology, origin, form and distribution of nondispersive protein bodies are discussed. Their ultrastructural composition is described as being predominantly spindle-shaped, compound- spherical, rod-shaped and rosette-like. Based on the data accumulated from over 450 species (of about 3000 screened) it is evident that their taxonomic range is confined to a few dicotyledon superorders. Compound-spherical nondispersive protein bodies are characteristic of most of the Malvanae/Violanae; spindle-shaped forms are restricted to the Fabaceae (Rutanae). Rosanae-Proteanae-Myrtanae and the Magnolianae are the only other superorders that contain nondispersive protein bodies in several of their families. Evolutionary trends and possible taxonomic consequences implied in this distribution are discussed.

Of all the ultrastructural features recognized within sieve elements their specific plastids provide the most successful characters in seed plant systematics. Sieve–element plastids are classified into subtypes and forms according to their protein and starch contents. Presently 26 different forms grouped into six subtypes within the two basic types (S– and P–) may be discerned. Plastid forms containing protein crystals, protein filaments and starch grains are proposed to be primitive, forms missing any one of these contents held to be derived (a synoptical key is given to ease the identification of the different plastid forms). Based on the quantitative distribution of over 1500 investigated species and the suggested evolution of plastid forms a cladistic diagram is prepared to demonstrate interrelationships between forms of sieve–element plastids and the evolution of seed plant taxa. Correlations exist between subtype PII and Monocotyledons, subtype PHI and Centrospermae, subtype PIV and Fabales. Genuine plastid forms characterize Buxaceae (PVIc), Erythroxylaceae and Rhizophoraceae (both PVc) and Cyrillaceae (PVcf). The Magnoliiflorae are distinct by presence in its families of a great number of forms of subtype PI. Phylogenetic correlations for some of these taxa are discussed. – Crystalline P–protein of sieve–elements provides another character to be used for the delimitation of some families (e.g. Fabaceae), while presently ER–complexes or other organelles of sieve elements do not contribute to seed plant systematics.

Sieve-element plastids may contain any combination of protein crystals (c), protein filaments (f), and starch grains (s), or none of these. All structurally distinct combinations (=forms) possible (s, cs, cfs, c, cf, fs, f, o) are recorded among the 2100 dicotyledons investigated so far with the transmission electron microscope (representing 381 of more than 460 families described). The six forms that include c and/or f define the P-type and are found in some 620 species, mainly confined to Magnolianae, Caryophyllanae, Vitanae, and Rutanae (Fabales, Linales, Rhizophorales). A detailed family-by-family analysis and quantitative form-by-form comparison suggests that form-Ss plastids give rise to all other forms. Based on a logic model connecting all forms via one-step-alterations (loss or gain of starch, protein crystals, or filaments), the distribution of P-type plastids within dicotyledons, their interconnection, and their relationships to the S-type taxa are discussed. Due to the diversity of sieve-element plastids found within one taxon, the Magnolianae (Pand S-type) are held to be a center of P-type evolution and more primitive than Caryophyllanae (P-type, only) or other superorders. The significance of sieve-element plastid data to the shaping of some of the higher dicotyledon taxa is discussed.

Based on TEM investigations of some 1850 species and SEM examinations of about 6000 species of the Angiospermae, this is a survey of ultrastructural and micromorphological data (excluding pollen wall characters) which contribute valuable information for the classification of angiosperms. TEM characters predominately relate to phloem features, such as sieve–element plastids and crystalline P–protein, and to those equally present in other tissues, e.g. nuclear protein crystals and dilated ER–cisternae (DC). Of these, the sieve–element plastids with their types and subtypes (S, PI–PVI) and their over 20 forms represent the most thoroughly investigated TEM character. SEM characters mainly relate to epidermal surface features and can be grouped into four categories: (1) Cellular arrangement or cellular pattern; (2) Shape of cells (the “primary sculpture” of a surface); (3) Relief of outer cell walls (“the secondary sculpture” superimposed on the primary sculpture), caused mainly by cuticular striations and superficially visible wall inclusions and wall thickenings; (4) Epicuticular secretions (the “tertiary sculpture” superimposed on the secondary sculpture), i.e. mainly waxes and related substances. Ultrastructural evidence from sieve–element plastids for the classification of Mag–noliiflorae, Caryophylliflorae, Fabiflorae and the unity of the Monocotyledoneae is discussed, while further plastid data are listed for single families (e.g. Buxaceae, Cyrillaceae, Erythroxylaceae, Eucryphiaceae, Gunneraceae, Rhizophoraceae, Vitaceae). Crystalline P–protein dominates in Malviflorae, Violiflorae and Fabiflorae. Nuclear protein crystals are a specific feature of sieve elements of Boraginaceae. DC characterize Capparales s.lat. Micromorphological evidence derived from specific trichomes is presented as an aid to the characterization of Urticales and Loasales, while a detailed analysis of the micromorphology of the seed coat of Cactaceae and Orchidaceae provided new information for the classification of these families at the tribal and generic levels. As a completely new systematic feature for the classification of the Monocotyledoneae first results of micromorphological differences in wax crystalloids and their orientation patterns are presented: The Liliiflorae s. str. are clearly separated against the Zingiberiflorae–Commeliniflorae (incl. Velloziales, Bromeliales, Typhales) and Areciflorae, both characterized by two mutually exclusive and very specific wax types and delimited against taxa with unspecific waxes in the rest of the monocotyledons and all dicotyledons.