Holoparasitic Rafflesiaceae possess the most reduced endophytes and yet give rise to the world's largest flowers.

BACKGROUND AND AIMS: Species in the holoparasitic plant family Rafflesiaceae exhibit one of the most highly modified vegetative bodies in flowering plants. Apart from the flower shoot and associated bracts, the parasite is a mycelium-like endophyte living inside their grapevine hosts. This study provides a comprehensive treatment of the endophytic vegetative body for all three genera of Rafflesiaceae (Rafflesia, Rhizanthes and Sapria), and reports on the cytology and development of the endophyte, including its structural connection to the host, shedding light on the poorly understood nature of this symbiosis. METHODS: Serial sectioning and staining with non-specific dyes, periodic-Schiff's reagent and aniline blue were employed in order to characterize the structure of the endophyte across a phylogenetically diverse sampling. KEY RESULTS: A previously identified difference in the nuclear size between Rafflesiaceae endophytes and their hosts was used to investigate the morphology and development of the endophytic body. The endophytes generally comprise uniseriate filaments oriented radially within the host root. The emergence of the parasite from the host during floral development is arrested in some cases by an apparent host response, but otherwise vegetative growth does not appear to elicit suppression by the host. CONCLUSIONS: Rafflesiaceae produce greatly reduced and modified vegetative bodies even when compared with the other holoparasitic angiosperms once grouped with Rafflesiaceae, which possess some vegetative differentiation. Based on previous studies of seeds together with these findings, it is concluded that the endophyte probably develops directly from a proembryo, and not from an embryo proper. Similarly, the flowering shoot arises directly from the undifferentiated endophyte. These filaments produce a protocorm in which a shoot apex originates endogenously by formation of a secondary morphological surface. This degree of modification to the vegetative body is exceptional within angiosperms and warrants additional investigation. Furthermore, the study highlights a mechanical isolation mechanism by which the host may defend itself from the parasite.

Partial shoot reiteration in Wollemia nobilis (Araucariaceae) does not arise from 'axillary meristems'.

BACKGROUND AND AIMS: Conifers are characterized by the paucity of axillary buds which in dicotyledonous trees usually occur at every node. To compensate, conifers also produce 'axillary meristems', which may be stimulated to late development. In juvenile material of Wollemia nobilis (Araucariaceae: Massart's model) first-order (plagiotropic) branches lack both axillary buds and, seemingly, axillary meristems. This contrasts with orthotropic (trunk) axes, which produce branches, either within the terminal bud or as reiterated orthotropic axes originating from axillary meristems. However, plagiotropic axes do produce branches if they are decapitated. This study investigated how this can occur if axillary meristems are not the source. METHODS: The terminal buds of a series of plagiotropic branches on juvenile trees were decapitated in order to generate axillary shoots. Shoots were culled at about weekly intervals to obtain stages in lateral shoot development. Serial sections were cut with a sliding microtome from the distal end of each sample and scanned sequentially for evidence of axillary meristems and early bud development. KEY RESULTS: Anatomical search produced no clear evidence of pre-existing axillary meristems but did reveal stages of bud initiation. Buds were initiated in a group of small starch-rich cortical cells. Further development involved de-differentiation of these small cells and the development of contrasting outer and inner regions. The outer part becomes meristematic and organizes the apex of the new branch. The inner part develops a callus-like tissue of vacuolated cells within which vascular cambia are developed. This kind of insertion of a branch on the parent axis seems not to have been described before. CONCLUSIONS: Axillary meristems in Wollemia characterize the leaf axils of trunk axes so that the origin of reiterated shoots is clear. Plagiotropic axes seemingly lack axillary meristems but still produce axillary branches by distinctive developmental processes. These observations demonstrate limited understanding of branch initiation in trees generally.

Stem vascular architecture in the rattan palm Calamus (Arecaceae-Calamoideae-Calaminae).

Climbing stems in the rattan genus Calamus can reach lengths of well over 100 m, are long-lived, and yet their vascular tissue is entirely primary. Such a combination suggests that stem vasculature is efficient and resistant to hydraulic disruption. By means of an optical shuttle and video recording of sequential images we analyzed the stem of a cultivated species. The stem has vascular features that are unusual compared with those in arborescent palms and seemingly inefficient in terms of long-distance water transport. Axial bundles are discontinuous basally because leaf traces, when followed downwards, always end blindly below. Furthermore, there is no regular distal branching of each leaf trace at its level of departure into a leaf, so that neither a continuing axial bundle nor bridges to adjacent axial bundles are produced as in the standard palm construction. Instead, the axial bundles in the stem periphery are connected to leaf traces and to each other by narrow and irregular transverse or oblique commissures that are not the developmental homologues of bridges. As in other palms, metaxylem within a leaf trace is not continuous into the leaf so that the only connection to a leaf is via protoxylem. Within the stem, protoxylem (tracheids) and metaxylem (vessels) are never contiguous, unlike in other palms, which suggests that water can only move from metaxylem to protoxylem, and hence into the leaf, across a hydraulic resistance. We suggest that this minimizes cavitation of vessels and/or may be associated with an unknown mechanism that refills embolized vessels. Also, the metaxylem can be significant in stem water storage in the absence of abundant ground parenchyma.

Apical dichotomy demonstrated in the angiosperm flagellaria.

An equal dichotomy of an angiosperm vegetative apex, previously suspected on morphological grounds, is demonstrated both externally in whole apices and in histological preparations. Dichotomy is a normal developmental feature with complete continuity in growth from the simple to the bifurcated state. The accessibility of this rare phenomenon in this plant makes further, possibly experimental, approaches feasible.

Analysis of complex vascular systems in plants: optical shuttle method.

The "optical shuttle" is an improved method of photographing with a motion-picture camera, one by one, sequential sections mounted on separate slides. The optical output from two microscopes is combined so that the images can be focused on a single film plane. Slides are photographed alternately through the microscopes. Simultaneous viewing of two successive sections in the microscopes enables initial precise alignment of images. Complex anatomical structures can thus be analyzed with relative ease.