Ultrastructure of pit membrane dissolution and movement of Xylella fastidiosa through pit membranes in petioles of Vitis vinifera

Xylella fastidiosa is the causative agent in Pierce’s disease (PD) in Vitis vinifera L. (grape) vines. Xylella fastidiosa colonizes and disseminates itself from one xylem element to another by dissolution and breach of pit membranes. These studies on naturally infected V. vinifera grown under vineyard conditions document by transmission electron microscopy that there is dissolution and breach of pit membranes by X. fastidiosa in vertical and lateral colonization in PD. These processes were documented in two cultivars of V. vinifera: ‘Syrah’ and ‘Cabernet Sauvignon.’

Elemental Analysis of Phloem Tissue in Lemna Minor Root Tips

Studies (1—4) have shown that it is possible to distinguish different stages of phloem tissue differentiation in the developing roots of Lemna minor by examination in the transmission, scanning, and optical microscopes. A disorganized meristem, immediately behind the root-cap, gives rise to the vascular tissue, which consists of single central xylem element surrounded by a ring of phloem parenchyma cells. This ring of cells is first seen at the 4-5 cell stage, but increases to as many as 11 cells by repeated radial anticlinal divisions. At some point, usually at or shortly after the 8 cell stage, two phloem parenchyma cells located opposite each other on the ring of cells, undergo an unsynchronized, periclinal division to give rise to the sieve element and companion cell. Because of the limited number of cells involved, this developmental sequence offers a relatively simple system in which some of the factors underlying cell division and differentiation may be investigated, including the distribution of diffusible low atomic weight elements within individual cells of the phloem tissue.

ATPase in mature and differentiating phloem and xylem.

A cytochemical study using a lead precipitation technique has been made of the distribution of adenosine triphosphatase (ATPase) in mature and differentiating phloem and xylem cells of Nicotiana tabacum and Pisum sativum. The sites of ATPase localization in tobacco phloem were the plasma membrane, endoplasmic reticulum, mitochondria, dictyosomes, plasmodesmata, and the dispersed P proteins of mature sieve elements. In pea phloem sieve elements ATPase was localized in the endoplasmic reticulum, but was not associated with the P proteins or plasma membranes at any stage of their differentiation. In pea transfer cells ATPase activity was associated with the endoplasmic reticulum at all stages of their differentiation and with the plasma membrane of transfer cells that had formed wall ingrowths. In xylem cells of both tobacco and pea the patterns of ATPase activity was similar. At early stages of differentiation ATPase activity was associated with the plasma membrane and the endoplasmic reticulum. At intermediate stages of differentiation ATPase activity continued to be associated with the endoplasmic reticulum, but was no longer associated with the plasma membrane. At later stages of xylem element differentiation ATPase activity was associated with disintegrating organelles and with the hydrolyzing cell walls.

THE DEVELOPMENT OF THE SECONDARY WALL OF THE XYLEM IN ACER PSEUDOPLATANUS

The development of the spirally thickened xylem element from a cambium initial of sycamore Acer pseudoplatanus has been traced by means of electron microscopy. The narrow elongated cambial initial undergoes considerable expansion in all dimensions. The cytoplasm at this stage is distributed in a thin skin between the cell wall and a large vacuole. No correlation has been observed between the distribution of any organelle and the pattern of the eventual thickenings. After the sites of thickening deposition have become apparent, the most conspicuous feature of the cell is the proliferation of Golgi bodies and vesicles. It is suggested that the material of the developing thickenings stems from direct apposition of the material in the Golgi vesicles. After glutaraldehyde fixation, microtubules (200 to 220 A in diameter) are seen to be sited in specific relation to the thickenings, the orientation of the tubules mirroring that of the fibrils seen in the thickenings. Possible reasons for absence of an observable pattern in the expanded but relatively undifferentiated cell are given, and the possible roles of the Golgi apparatus and microtubules in the thickening production are discussed