An Aphid-Secreted Salivary Protease Activates Plant Defense in Phloem

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Uptake,Transport and Storage of Cardenolides in Foxglove. Cardenolide Sinks and Occurrence of Cardenolides in the Sieve Tubes of Digitalis lanata1

Cardiac glycoside transport was investigated on the organ and whole plant level. Uptake experiments were carried out with shoot and root cultures of Digitalis lanata. In both systems primary cardenolides, i.e., those with a terminal glucose in their oligosaccharide side chain, were taken up against their concentration gradient, whereas the glucose-free secondary cardenolides were not. Active uptake of primary cardenolides was further evidenced by KCN inhibition of uptake. Using plantlets grown in vitro the long-distance transport of primary cardenolides from the leaves to the roots was demonstrated. Cardenolides were also detected in etiolated leaves, induced on plants with green leaves, which are supposed to be unable to synthezise cardenolides de novo, providing further evidence for long-distance transport. Several primary cardenolides were detected in the honeydew excreted by aphids fed on Digitalis lanata leaves, indicating that the phloem is a transporting tissue for cardenolides. On the other hand, the xylem sap obtained by applying the pressure-chamber technique was cardenolide-free. It was concluded that in Digitalis primary cardenolides serve as both the transport and the storage form of cardenolides. After their synthesis they are either stored in the vacuoles of the source tissue or loaded into the sieve tubes, from which they are unloaded at other sites where they are trapped in the vacuoles of the respective sink tissue.     

Translocation - A key factor limiting the efficiency of nitrogen fixation in legume nodules

A hypothesis is presented that the availability of water for export of nitrogenous products from legume nodules is a major factor limiting the efficiency of symbiotic nitrogen fixation. Water for export of solutes in the xylem probably depends largely on the import of water and reduced carbon in the phloeum, and one function of respiration may be to dispose of reduced carbon in order to increase the supply of water. A second hypothesis presented is that control of gas diffusion in soybean nodules is largely restricted to the cortex nearby the vascular bundles, thus making possible the linkage of solute balances in xylem and phloem with resistance to diffusion. These concepts are used in a re-examination of literature on manipulations of nodules and nodulated plants such as lowering of light levels, water stress, defoliation, stem girdling, and alteration of oxygen supply. The concept of translocation as a major factor limiting efficiency of symbiotic fixation is consistent with the failure of superior rhizobial isolates to improve N input significantly, and this limitation could also prevent exploitation of superior bacterial symbionts in the future     

Honeydew amino acids in relation to sugars and their role in the establishment of ant-attendance hierarchy in eight species of aphids feeding on tansy (Tanacetum vulgare)

Abstract. The ratio of the concentration of honeydew total amino acids to total sugars in the honeydew of eight species of aphids, all feeding on tansy, Tanacetum vulgare (L.), was determined and correlated with honeydew production and ant‐attendance. The honeydew of the five ant‐attended aphid species [Metopeurum fuscoviride (Stroyan), Trama troglodytes (v. Hayd), Aphis vandergooti (Börner), Brachycardus cardui (L.), Aphis fabae (Scopoli)] was rich in total amino acids, ranging from 12.9 to 20.8 nmol µL^?1 compared with the unattended aphid Macrosiphoniella tanacetaria (Kalt.) with only 3 nmol µL^?1. Asparagine, glutamine, glutamic acid and serine (all nonessential amino acids) were the predominant amino acids in the honeydew of all species. The total concentration of amino acids in the phloem sap of tansy was much higher (78.7 nmol µL^?1) then in the honeydew samples, and the predominant amino acids were glutamate (34.3%) and threonine (17.7%). A somewhat unexpected result was the finding that those aphid species with the highest total amino acid concentration in the honeydew always had the highest concentration of sugars. The lowest amino acid–sugar combined value was 104–28.8 nmol µL^?1 in the non ant‐attended species M. tanacetaria, and the highest value was an average of 270–89.9 nmol µL^?1 for the three most intensely attended aphid species M. fuscoviride, A. vandergooti and T. troglodytes. There is no evidence that any single amino acid or group of amino acids in the honeydew acted as an attractant for ant‐attendance in these eight aphid species. The richness of the honeydew (rate of secretion × total concentration of sugars), along with the presence of the attractant sugar melezitose, comprised the critical factors determining the extent of ant‐attendance of the aphids feeding on T. vulgare. The high total amino acid concentration in sugar‐rich honeydews can be explained by the high flow‐through of nutrients in aphids that are particularly well attended by ants.     

Sucrose transport into the phloem of Ricinus communis L. seedlings as measured by the analysis of sieve-tube sap

Careful cutting of the hypocotyl of Ricinus communis L. seedlings led to the exudation of pure sieve-tube sap for 2–3 h. This offered the possibility of testing the phloem-loading system qualitatively and quantitatively by incubating the cotyledons with different solutes of various concentrations to determine whether or not these solutes were loaded into the sieve tubes. The concentration which was achieved by loading and the time course could also be documented. This study concentrated on the loading of sucrose because it is the major naturally translocated sieve-tube compound. The sucrose concentration of sieve-tube sap was approx. 300 mM when the cotyledons were buried in the endosperm. When the cotyledons were excised from the endosperm and incubated in buffer, the sucrose concentration decreased gradually to 80–100 mM. This sucrose level was maintained for several hours by starch breakdown. Incubation of the excised cotyledons in sucrose caused the sucrose concentration in the sieve tubes to rise from 80 to 400 mM, depending on the sucrose concentration in the medium. Thus the sucrose concentration in the sieve tubes could be manipulated over a wide range. The transfer of labelled sucrose to the sieve-tube sap took 10 min; full isotope equilibration was finally reached after 2 h. An increase of K⁺ in the medium or in the sieve tubes did not change the sucrose concentration in the sievetube sap. Similarly the experimentally induced change of sucrose concentration in the sieve tubes did not affect the K⁺ concentration in the exudate. High concentrations of K⁺, however, strongly reduced the flow rate of exudation. Similar results were obtained with Na⁺ (data not shown). The minimum translocation speed in the sieve tubes in vivo was calculated from the growth increment of the seedling to be 1.03 m·h^-1, a value, which on average was also obtained for the exudation system with the endosperm attached. This comparison of the in-vivo rate of phloem transport and the exudation rate from cut hypocotyls indicates that sink control of phloem transport in the seedlings of that particular age was small, if there was any at all, and that the results from the experimental exudation system were probably not falsified by removal of the sink tissues.Abbreviations PTS 3-hydroxy-5,8, 10-pyrenetrisulfonate     

Current perspectives on plasmodesmata: structure and function

Recent studies on plasmodesmata have shown that these important intercellular passages for communication and transport are much more sophisticated in both structure and regulatory abilities than previously imagined. A complex, but not well understood, substructure has been revealed by a variety of increasingly reliable ultrastructural techniques. Proteinaceous particles are seen within the cytoplasmic sleeve surrounding the desmotubule. Dye-coupling studies have provided experimental evidence for the physical pathway of solute movement, supporting conclusions about substructural dimensions within plasmodesmata drawn from the ultrastructural studies. Calcium has been identified as a major factor in the regulation of intercellular communication via plasmodesmata. Evidence from studies on virus movement through plasmodesmata suggests a direct interaction between virallycoded movement proteins and plasmodesmata in the systemic spread of many viruses. There is increasing evidence, albeit indirect, that in some plant species phloem loading may involve transport of photoassimilate entirely within the symplast from mesophyll cells to the sieve element-companion cell complexes of minor veins.     

Resistance of different lucerne cultivars to the pea aphid Acyrthosiphon pisum: influence of phloem composition on aphid fecundity

Sugars and amino acids were analysed in the phloem sap (sampling by the exudation method) of four clones of lucerne (Medicago sativa L.) characterized by their resistance to the pea aphid, Acyrthosiphon pisum (Harris) (Homoptera: Aphididae). Differences in the qualitative amino acid balance are shown and a negative correlation between the reproductive rate of the aphid and the sugars/amino acids ratio is underlined. To demonstrate any causal relationship between these two variables, the reproduction of A. pisum on artificial diets copying the sap of the two extreme clones (resistant and susceptible) was measured. The results show that the resistance is not a simple nutritional effect; however, the amino acid balance contributes to the resistance exhibited by some cultivars.

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Résumé L'analyse (sucres et acides aminés) de la sève de 4 cultivars de luzerne (Medicago sativa) caractérisés pour leur résistance au puceron du pois (Acyrthosiphon pisum (Harris)) (Homoptera: Aphididae), est réalisée sur des exsudats de phloème. La teneur en sucres (exprimée en équivalent saccharose) montre d'importantes différences: rapport de 1 à 3 respectivement entre les clones sensible et résistant. La teneur globale en acides aminés exsudés varie en sens inverse, entraînant une corrélation négative pour les 4 clones étudiés entre le rapport sucre/acides aminés (0,6 à 3,6) et la résistance mesurée par le taux net de reproduction à 14 jours (78,5 à 11,2 larves/femelle). L'analyse des acidses aminés révèle de plus des différences qualitatives entre cultivars, notamment pour certains acides aminés essentiels méthionine, histidine, lysine, aromatiques.Le rôle de l'équilibre en acides aminés et en sacharose dans la résistance de certains cultivars au puceron du pois est apprécié par des mesures de la reproduction de A. pisum sur des milieux artificiels reproduisant le rapport sucre/acides aminés et/ou le spectre des acides aminés des 2 clones extrêmes (sensible et résistant). La résistance n'apparaît pas être le résultat d'une simple différence de la qualité nutritionnelle de la sève; la balance en acides aminés doit cependant participer pour une part à la résistance au puceron.

     

Age-specific flight behavior in relation to body weight and lipid content of Ips calligraphus reared in slash pine bolts with thick or thin inner bark (phloem)

Rearing larvae of Ips calligraphus (Germar) (Coleoptera: Scolytidae) in bolts of typical slash pine, Pinus elliottii Engelm. var. elliottii, with thin (TN; 0.5–1.5 mm) phloem (inner bark) relative to adult beetle body width significantly reduced body weight and lipid content of emerging adults compared with thick (TK; 2.5–3.5 mm) phloem-reared adults. Unmated (pre-nuptial) TK beetles averaged 2-fold longer daily and life-time flight duration than TN beetles. Daily flights were highly variable, averaging <20 min (longest daily flight=173 min). TK and TN beetles initiated flight, and the greatest number flew, on the first or second day after emergence form their rearing bolt; % of beetles flying and survival gradually declined to zero by day 9. Males (both TK and TN) averaged 2.5 days of flight, ca. 1 day longer than females. Fliers lived ca. 2-fold longer than non-fliers. Among TK and TN fliers, significant but weak positive correlations occurred between body weight and lifetime flight duration. Poor correlation between lifetime flight duration and longevity suggests little physiological cost to flight, but effects on fecundity were not assessed. Flight by mated, egg-laying (post-nuptial) beetles was substantially delayed (peaking on day 6), suggesting degeneration and subsequent regeneration of flight muscles.

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Résumé Le comportement de vol d'Ips calligraphus GERMAR (Scolytidae) a été examiné au laboratoire par une technique de vol captif. La génération parentale avait été artificiellement introduite, suivant la taille des adultes (1,6 à 2,1 mm) dans des morceaux de Pinus elliottii Engelm. variété elliottii au phloeme épais (TK=2,5 à 3,5 mm) ou fin (TN=0,5 à 1,5 mm) les larves se sont développées dans ces morceaux de pin. Le poids et la teneur en lipide des adultes obtenus à partir de TN étaient significativement inférieure à ceux de TK. Le poids des mâles les plus gros a été plus réduit que celui des femelles; les mâles les plus petits semblaient être moins efficaces dans l'initiation des attaques sur les arbres, et les femelles les plus petites ont eu une fécondité réduite.Les durées de vol quotidiennes pendant la vie des mâles TK vierges étaient en moyenne deux fois plus longues que celles des mâles TN. Les durées de vol quotidiennes étaient très variables: en moyenne moins de 20 minutes, mais la plus longue a été de 173 mn. En moyennes les adultes TN et TK ont commencé à voler, et volaient en plus grand nombre, le premier et le second jour après l'émergence. Ensuite la fréquence d'adultes volants et le pourcentage de surviveants, ont graduellement diminué jusqu'à tomber à zéro le neuvième jour. Les mâles TN et TK avaient des vols quotidiens plus longs et volaient environ 1 jour de plus que les femelles. L'activité de vol accrue des mâles reflète leur besoin de trouver dans la nature des arbres convenables; les vols des femelles peuvent généralement être plus brefs, étant plus directement orientés comme une réponse aux phéromones mâles.Les voiliers ont vécu environ 2 fois plus longtemps que les non-voiliers. Parmi les voiliers TK et TN il y avait une corrélation positive étroite entre le poids du corps et la durée totale des vols. Il n'y en a qu'une faible corrélation entre cette durée totale des vols et la longévité, suggérant un faible coût physiologique du vol. L'influence de cette durée des vols et du prélèvement consécutif de lipides sur la fécondité n'a pas été examinée. L'initiation au vol chez les individus fécondés a été sérieusement retardée (maximum le sixième jour) par rapport aux vierges, ce qui suggère un dégénérescence des muscles du vol associée à la reproduction, suivie d'une régénération importante, comme cela a été signalé chez d'autres Scolytes.Ces résultats contribuent à notre compréhension des facteurs conditionnant le comportement de vol et d'autres aspects fondamentaux de la biologie des insectes des pins. Une telle connaissance est nécessaire au développement d'une politique efficace de protection.

     

Import of 14C-Photosynthate by Developing Leaves of Sugarcane

Sink-to-source transition was studied in developing sugarcane (Saccharum interspecific variety L62–96) leaves. Fully-expanded, mature sugarcane leaves were fed ¹⁴CO₂ for 20 minutes, incorporating about 617 Bq. After five hours the leaves of each plant were cut into 1-cm-length segments that were weighed and then placed in scintillation cocktail for counting. All leaves younger than the leaf fed ¹⁴CO₂ imported labeled photoassimilate. Three to four leaves had both importing and non-importing regions within the blade and a distinct transition region between them. A transition region was observed in leaves which had expanded to between 30 and 90 % of final blade length. Radioactivity per gram fresh weight was calculated as a measure of sink strength. Sink strength was greatest in the youngest leaf and declined with leaf age. The results of this study indicate that 1) import of photosynthate by developing sugarcane leaves occurs over a longer span of developmental ages than in dicotyledonous leaves and 2) the actual tissue region undergoing transition within such a leaf can be resolved as narrow zone between the importing and non-importing regions.