The life of phi: the development of phi thickenings in roots of the orchids of the genus Miltoniopsis.

Phi thickenings, bands of secondary wall thickenings that reinforce the primary wall of root cortical cells in a wide range of species, are described for the first time in the epiphytic orchid Miltoniopsis. As with phi thickenings found in other plants, the phi thickenings in Miltoniopsis contain highly aligned cellulose running along the lengths of the thickenings, and are lignified but not suberized. Using a combination of histological and immunocytochemical techniques, thickening development can be categorized into three different stages. Microtubules align lengthwise along the thickening during early and intermediate stages of development, and callose is deposited within the thickening in a pattern similar to the microtubules. These developing thickenings also label with the fluorescently tagged lectin wheat germ agglutinin (WGA). These associations with microtubules and callose, and the WGA labeling, all disappear when the phi thickenings are mature. This pattern of callose and WGA deposition show changes in the thickened cell wall composition and may shed light on the function of phi thickenings in plant roots, a role for which has yet to be established.

The Orchid Velamen: A Model System for Studying Patterned Secondary Cell Wall Development?

Understanding the mechanisms through which plants generate secondary cell walls is of more than academic interest: the physical properties of plant-derived materials, including timber and textiles, all depend upon secondary wall cellulose organization. Processes controlling cellulose in the secondary cell wall and their reliance on microtubules have been documented in recent decades, but this understanding is complicated, as secondary walls normally form in the plant's interior where live cell imaging is more difficult. We investigated secondary wall formation in the orchid velamen, a multicellular epidermal layer found around orchid roots that consists of dead cells with lignified secondary cell walls. The patterns of cell wall ridges that form within the velamen vary between different orchid species, but immunolabelling demonstrated that wall deposition is controlled by microtubules. As these patterning events occur at the outer surface of the root, and as orchids are adaptable for tissue culture and genetic manipulation, we conclude that the orchid root velamen may indeed be a suitable model system for studying the organization of the plant cell wall. Notably, roots of the commonly grown orchid Laelia anceps appear ideally suited for developing this research.

The Induction and Roles Played by Phi Thickenings in Orchid Roots.

Phi thickenings are specialised secondary wall thickenings present in the root cortex of many plant species, including both angiosperms and gymnosperms. While environmental stresses induce phi thickenings, their role(s) in the root remain unclear. Suggested functions include regulation of transport through the apoplast in a manner similar to the Casparian strip, limiting fungal infections, and providing mechanical support to the root. We investigated phi thickening induction and function in Miltoniopsis sp., an epiphytic orchid. As movement of a fluorescent tracer through the apoplast was not blocked by phi thickenings, and as phi thickenings developed in the roots of sterile cultures in the absence of fungus and did not prevent fungal colonisation of cortical cells, the phi thickenings in Miltoniopsis did not function as a barrier. Phi thickenings, absent in roots grown on agar, remained absent when plants were transplanted to moist soil, but were induced when plants were transplanted to well-drained media, and by the application of water stress. We suggest that it is likely that phi thickenings stabilise to the root during water stress. Nevertheless, the varied phi thickening induction responses present in different plant species suggest that the phi thickenings may play multiple adaptive roles depending on species.