Transport of glutamine into the xylem of sunflower (Helianthus annuus).

Sunflower (Helianthus annuus L.) plants were grown on nutrient solution with ammonium nitrogen. After 12 days of growth the ammonium in the nutrient solution was labeled with N (99%). Three hours later glutamine-N in the xylem exudate was labeled for 56% as shown by GC-MS; this percentage increased to 63% after 8, and to 69% after 24 hours of incubation. When the xylem exudate had been collected from the epicotyl instead of the hypocotyl, 15-N abundances were 52%, 56% and 63% respectively. Results are consistent with an import of glutamine into the transpiration stream during its ascension in the xylem. On basis of the differences in abundance of double-labeled, single-labeled and unlabeled glutamine between the two sampling sites it was estimated that at least 20% of the xylem glutamine was imported into xylem along this distance (~4cm)

Overexpression of the aphid-induced serine protease inhibitor <i>CI2c </i>gene in barley affects the generalist green peach aphid, not the specialist bird cherry-oat aphid

<div><p>Aphids are serious pests in crop plants. In an effort to identify plant genes controlling resistance against aphids, we have here studied a protease inhibitor, CI2c in barley (<i>Hordeum vulgare</i> L.). The <i>CI2c</i> gene was earlier shown to be upregulated by herbivory of the bird cherry-oat aphid <i>(Rhopalosiphum padi</i> L.<i>)</i> in barley genotypes with moderate resistance against this aphid, but not in susceptible lines. We hypothesized that CI2c contributes to the resistance. To test this idea, cDNA encoding <i>CI2c</i> was overexpressed in barley and bioassays were carried out with <i>R</i>. <i>padi</i>. For comparison, tests were carried out with the green peach aphid (<i>Myzus persicae</i> Sulzer), for which barley is a poor host. The performance of <i>R</i>. <i>padi</i> was not different on the <i>CI2c</i>-overexpressing lines in comparison to controls in test monitoring behavior and fecundity. <i>M</i>. <i>persicae</i> preference was affected as shown in the choice test, this species moved away from control plants, but remained on the <i>CI2c</i>-overexpressing lines. <i>R</i>. <i>padi</i>-induced responses related to defense were repressed in the overexpressing lines as compared to in control plants or the moderately resistant genotypes. A putative susceptibility gene, coding for a β-1,3-glucanase was more strongly induced by aphids in one of the <i>CI2c</i>-overexpressing lines. The results indicate that the CI2c inhibitor in overexpressing lines affects aphid-induced responses by suppressing defense. This is of little consequence to the specialist <i>R</i>.<i>padi</i>, but causes lower non-host resistance towards the generalist <i>M</i>. <i>persicae</i> in barley.</p></div

Collection of phloem sap in phytoplasma-infected plants

Phytoplasmas colonize specifically the phloem sieve elements (SEs) of plants and influence effectively the plant physiology. To study and understand the interaction of phytoplasmas and host plants an access to the cellular, microscale volume of SEs is demanded. Different methods are suitable to collect phloem sap of phytoplasma-infected plants. The two most common methods are the EDTA-facilitated exudation and the stylectomy. For the EDTA-facilitated method, the cut end of a leaf is placed into an EDTA solution. The EDTA prevents and avoids the Ca2+ dependent (re-) occlusion of SEs by binding Ca2+ ions and the mass flow of SEs is restarted which results in an outflow of the SE content into the EDTA bathing solution. The advantage is on the one hand a simple application and secondly, feasible for all plant species. The stylectomy method requires piercing-sucking insects like any aphids. During phloem-sap ingestion, the stylet is severed by a microcautery device or a laser from the insect body. Due to the high turgor pressure of the SEs the phloem sap is forced out through the remaining stylet and can be collected with a glass capillary, for example. The stylectomy delivers pure phloem sap, however, the collected volumes are in the range of nano liters and the temporal and staff costs are tremendous. A third method is the spontaneous exudation in phytoplasma-infected apple trees providing only in springtime large volumes of vascular sap after cutting along the bark. For the spontaneous exudation the proportion of phloem sap is unclear. Thus, this third method still needs a closer examination in prospective surveys