Optimal Preparation of Tissue Sections for Light-Microscopic Analysis of Phloem Anatomy.

In order to successfully analyze and describe any plant tissue, the first challenge is preparation of good anatomical slides. The challenge is even greater when the target tissue has heterogeneous characteristics, such as the phloem where soft and stiff tissues occur side by side. The goal of this chapter is to present a detailed protocol containing various techniques for optimal preparation of phloem tissue samples for light microscopic analysis. The process typically involves the steps of fixation, softening, embedding, sectioning, staining, and mounting. The protocol can be applied to make samples of phloem and surrounding tissues of stems and roots, from woody to herbaceous plants.

Leaf emergences in Microlepis oleaefolia (DC.) Triana (Melastomataceae) and their probable function: an anatomical and ultrastructural study.

Microlepis oleaefolia (DC.) Triana, an endemic species of Melastomataceae from the Brazilian cerrado, presents very complex leaf structures called as "hairs with root-like base" embedded in the mesophyll. This paper describes the ontogenesis, histochemical and structural aspects of these leaf structures as a framework for further functional studies. Samples of leaves in different developmental stages were processed according to common light and electron microscopy techniques. Fresh material was subjected to histochemical tests to examine the chemical composition of cell walls. The apoplastic transport between leaf emergences and the vascular system was verified by staining with 1% aqueous safranin. The structures are emergences of mixed protoderm and ground meristem origin. They are persistent, predominantly short-stalked and dendritic. Their arms have thick non-lignified cellulosic walls with a loose and heterogeneous aspect; on the inner face, the wall, which appears labyrinthine, presents small irregularly shaped projections directed towards the protoplast. The base of the emergences is composed of sclereids embedded in the mesophyll that reach the vascular system. Assays with aqueous safranin solution revealed it penetrates the cell walls of the arms and showed a connection between the emergence and xylem. Anatomical, chemical and ultrastructural features of leaf emergences of M. oleaefolia indicate that these structures are able to transport substances via apoplast and can absorb or exude solutions.

Pulvinus functional traits in relation to leaf movements: a light and transmission electron microscopy study of the vascular system.

Previous studies on legume pulvini suggest that the vascular system plays an important role in the redistribution of ions and transmission of stimuli during leaf's movements. However, the number of anatomical and ultrastructural studies is limited to few species. The aim of this paper is to investigate the structure and cellular features of the pulvinus vascular system of nine legume species from Brazilian cerrado, looking for structural traits pointing to its participation in the leaf's movements. Samples were excised from the medial region of opened pulvinus of Bauhinia rufa, Copaifera langsdorffii, Senna rugosa (Caesalpinioideae), Andira humilis, Dalbergia miscolobium, Zornia diphylla (Faboideae), Mimosa rixosa, Mimosa flexuosa and Stryphnodendron polyphyllum (Mimosoideae), and were prepared following light microscopy, transmission electron microscopy and histochemical standard techniques. The vascular system occupies a central position, comprises phloem and xylem and is delimited by a living sheath of septate fibers in all the species studied. This living cells sheath connects the cortex to the vascular tissues via numerous plasmodesmata. The absence of fibers and sclereids, the presence of phenolic idioblasts and the abundance and diversity of protein inclusions in the sieve tube members are remarkable features of the phloem. Pitted vessel elements, parenchyma cells with abundant cytoplasm and living fibriform elements characterize the xylem. The lack of lignified tissues and extensive symplastic continuity by plasmodesmata are remarkable features of the vascular system of pulvini of the all studied species.

Plant cell proliferation inside an inorganic host.

In recent years, much attention has been paid to plant cell culture as a tool for the production of secondary metabolites and the expression of recombinant proteins. Plant cell immobilization offers many advantages for biotechnological processes. However, the most extended matrices employed, such as calcium-alginate, cannot fully protect entrapped cells. Sol-gel chemistry of silicates has emerged as an outstanding strategy to obtain biomaterials in which living cells are truly protected. This field of research is rapidly developing and a large number of bacteria and yeast-entrapping ceramics have already been designed for different applications. But even mild thermal and chemical conditions employed in sol-gel synthesis may result harmful to cells of higher organisms. Here we present a method for the immobilization of plant cells that allows cell growth at cavities created inside a silica matrix. Plant cell proliferation was monitored for a 6-month period, at the end of which plant calli of more than 1 mm in diameter were observed inside the inorganic host. The resulting hybrid device had good mechanical stability and proved to be an effective barrier against biological contamination, suggesting that it could be employed for long-term plant cell entrapment applications.