Studies onArtemisia afra Jacq.: The phloem in stem and leaf

Summary The structure of the phloem was studied in stem and leaf ofArtemisia afra Jacq., with particular attention being given to the sieve element walls. Both primary and secondary sieve elements of stem and midvein have nacreous walls, which persist in mature cells. Histochemical tests indicated that the sieve element wall layers contained some pectin. Sieve element wall layers lack lignin. Sieve elements of the minor veins (secondary and tertiary veins) lack nacreous thickening, although their walls may be relatively thick. These walls and those of contiguous transfer cells are rich in pectic substances. Transfer cell wall ingrowths are more highly developed in tertiary than in secondary veins.     

Leaf structure in relation to solute transport and phloem loading in Zea mays L.

Small and intermediate (longitudinal) vascular bundles of the Zea mays leaf are surrounded by chlorenchymatous bundle sheaths and consist of one or two vessels, variable numbers of vascular parenchyma cells, and two or more sieve tubes some of which are associated with companion cells. Sieve tubes not associated with companion cells have relatively thick walls and commonly are in direct contact with the vessels. The thick-walled sieve tubes have abundant cytoplasmic connections with contiguous vascular parenchyma cells; in contrast, connections between vascular parenchyma cells and thin-walled sieve tubes are rare. Connections are abundant, however, between the thin-walled sieve tubes and their companion cells; the latter have few connections with the vascular parenchyma cells. Plasmolytic studies on leaves of plants taken directly from lighted growth chambers gave osmotic potential values of about-18 bars for the companion cells and thin-walled sieve tubes (the companion cell-sieve tube complexes) and about-11 bars for the vascular parenchyma cells. Judging from the distribution of connections between various cell types of the vascular bundles and from the osmotic potential values of those cell types, it appears that sugar is actively accumulated from the apoplast by the companion cell-sieve tube complex, probably across the plasmalemma of the companion cell. The thick-walled sieve tubes, with their close spatial association with the vessels and possession of plasmalemma tubules, may play a role in retrieval of solutes entering the leaf apoplast in the transpiration stream. The transverse veins have chlorenchymatous bundle sheaths and commonly contain a single vessel and sieve tube. Parenchymatic elements may or may not be present. Like the thick-walled sieve tubes of the longitudinal bundles, the sieve tubes of the transverse veins have plasmalemma tubules, indicating that they too may play a role in retrieval of solutes entering the leaf apoplast in the transpiration stream.     

The microscopy of P-protein filaments in freeze-etched sieve pores

Intact vascular bundles from Nymphoides peltata (S.G. Gmel.) O. Kuntze, shown to have translocated carbon-14, were freeze-fractured and etched for electron microscopy. The interpretation of freezefractured and etched sieve pores and P-protein filaments seen in them is discussed. The entire widths of most of the sieve pores seen contained filaments separated by less than 100 nm. Their arrangement indicates too high a resistance to flow for pressure flow alone to drive translocation at known rates; pumps would be necessary at places along sieve tubes. However, calculations are presented to show that during the time taken to fix pores, by fast freezing or chemically, the filaments in them could rearrange and move further by Brownian and other motion than the distances between filaments which we need to measure. These calculations show that it is not possible, by microscopy alone, to answer the outstanding question “How are filaments arranged in translocating sieve pores?” with enough certainty to tell us whether pressure flow is adequate to explain translocation where filaments are present. The calculations are relevant also to microscopy of other cell structures which may move.     

A guide to the use of the exuding-stylet technique in phloem physiology

The use of exuding stylets holds considerable promise for the investigation of sieve-tube physiology. However, largely because of difficulties in cutting insect stylets, the technique has been applied to only a few plant species. Based on our experience, a comparison is made of the available means of obtaining sieve-tube exudate from the exuding stylets of phloem-feeding insects, including aphids, scale and mealybugs. Forty-one plant species and approx. 35 insect species were tested for their ability to provide stylet exudate. Stylets on all but a few of the plant species tested yielded at least some exudate, but the success rate and duration of exudation on many species were unsatisfactory for detailed investigations of phloem transport. Plant species appears to be the most important factor for obtaining reliably exuding stylets, although the size of the insect species used and the physiological condition of the plant are also important variables. Armored scale provide a simple and reliable source of exuding stylets, but are impractical for most experimental purposes. Radio-frequency microcautery of aphid stylets was substantially the most effective means of cutting stylets. Instructions are provided for constructing a microcautery unit at minimal expense, using a citizen's band radio as the radio-frequency source.Abbreviations CB citizen's band - R.F. radio frequency     

Membrane receptor-mediated mechano-transduction maintains cell integrity during pollen tube growth within the pistil

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Ribosomes in incompatible pollen tubes in the Solanaceae

Some members of the Solanaceae have a self‐incompatibility mechanism preventing self‐fertilization. Stylar ribonucleases (S‐RNases) are responsible for growth inhibition of self‐pollen tubes. A prevalent model postulates that the S‐RNases act as intracellular cytotoxins that degrade ribosomal RNA, and possibly also messenger RNA, in the incompatible pollen tubes. Since ribosomes and polysomes are easily noticed with the electron microscope, it should be possible to confirm disintegration of these structures. However, our inspection by electron microscopy revealed the presence of ribosomes and polysomes in pollen tubes formed after self‐pollination of the self‐sterile species Brugmansia (Datura) suaveolens and Nicotiana alata . There was no decrease over time in the number of bound ribosomes per unit of rough endoplasmic reticulum (RER) membrane. The results indicate that the inhibition of tube growth is not due to a general degradation of ribosomal and messenger RNA. Therefore, the substrate for S‐RNases presumably is very specific.     

Restoration of a Forest Understory After the Removal of an Invasive Shrub, Amur Honeysuckle (Lonicera maackii)

The recruitment of native seedlings is often reduced in areas where the invasive Amur honeysuckle (Lonicera maackii) is abundant. To address this recruitment problem, we evaluated the effectiveness of L. maackii eradication methods and restoration efforts using seedlings of six native tree species planted within eradication and unmanipulated (control) plots. Two eradication methods using glyphosate herbicide were evaluated: cut and paint and stem injection with an EZ‐Ject lance. Lonicera maackii density and biomass as well as microenvironmental characteristics were measured to study their effects on seedling growth and survivorship. Mean biomass of Amur honeysuckle was 361 ± 69 kg/ha, and density was 21,380 ± 3,171 plants/ha. Both eradication treatments were effective in killing L. maackii (≥ 94%). The injection treatment was most effective on large L. maackii individuals (>1.5 cm diameter), was 43% faster to apply than cutting and painting and less fatiguing for the operator, decreased operator exposure to herbicide, and minimized impact to nontarget vegetation. Deer browse tree protectors were used on half of the seedlings, but did not affect survivorship or growth. After 3 years, survival of native seedlings was significantly less where L. maackii was left intact (32 ± 3%) compared with the eradication plots (p < 0.002). Seedling survival was significantly different between cut (51 ± 3%) and injected (45 ± 3%) plots. Species had different final percent survival and rates of mortality. Species survival differed greatly by species (in descending order): Fraxinus pennsylvanica > Quercus muehlenbergii ≥ Prunus serotina≥ Juglans nigra > Cercis canadensis > Cornus florida. Survivorship and growth of native seedlings was affected by a severe first‐year drought and by site location. One site exhibited greater spring soil moisture, pH, percent open canopy, and had greater survivorship relative to the other site (55 ± 2 vs. 30 ± 2%). Overall, both L. maackii eradication methods were successful, but restorationists should be aware of the potential for differential survivorship of native seedlings depending on species identity and microenvironmental conditions.     

Transmission of Electric Signals in Sieve Tubes of Zucchini Plants

Membrane potentials of ?;160 to ?210 mV were recorded with microelectrodes inserted into meta-phloem sieve tubes of intact zucchini plants (Cucurbita pepo L. var. medullosa Alef.). The effects of darkness, white light and colored light on membrane potential were studied. Reference electrodes were in contact with the apoplast via fluid-filled cavities or “drinks”. Electrolyte solutions (100 mM) in the cavities could be quickly replaced by flushing with 100 mM solutions of sucrose, KCl, sorbitol, or EDTA without altering osmolarity. KCl and EDTA caused depolarization of the sieve tube membrane potential, while sucrose caused depolarization or hyperpolarization of the sieve tube membrane potential in mature or growing plant parts respectively. Recovery of the original voltage was recorded when rapid (sucrose) or slow (sorbitol) transients occurred. When two measuring circuits were installed, one in a growing fruit and the other in the petiole of the subtending mature leaf, the alteration of the sieve tube membrane potential at one site was accompanied by an alteration of the potential at the other site after a few seconds. The responses were opposite in the exporting leaf and importing fruit when sucrose was applied. The signal, transmitted via the sieve tubes, reached maximum velocities of 10 cm per second.