Mobility of boron-polyol complexes in the hemiparasitic association between Rhinanthus minor and Hordeum vulgare: the effects of nitrogen nutrition.

Boron (B) is an essential nutrient required for plant growth and physiological processes. Long-distance B transport is facilitated by the formation of B-polyol complexes. We investigated B uptake and distribution in response to differing levels of exogenous nitrogen supply in the hemiparasitic association between Rhinanthus minor and Hordeum vulgare (barley) and in unparasitised barley and single Rhinanthus plants. In this system, the polyol mannitol is the major assimilate in Rhinanthus, whereas polyols are not detectable in barley. Furthermore, previous studies have shown that the accumulation of polyols within Rhinanthus is negatively affected by the application of exogenous nitrogen. Within the association, the strongest accumulation of B was detected in lateral buds and inflorescences of Rhinanthus, consistent with the greatest B demand in strong sink organs supplied through the phloem that contain high concentrations of mannitol. In the host, the strongest B accumulation was found in xylem-supported leaf lamellae. Roots and sheaths did not accumulate substantial amounts of B, while re-circulation of B through the phloem vessels accounted for only 10% (unparasitised) and 8% (parasitised) of the xylem sap-imported B in the mannitol-free barley hosts. In contrast, 53% (attached) and 39% (in the absence of a host) of the xylem sap-imported B was re-circulated in the phloem in the mannitol-rich Rhinanthus. We therefore present the first quantitative uptake and flow models of long-distance B transport in polyol-rich and polyol-free plants. Our findings are consistent with a close relationship between B re-translocation and mannitol concentrations in phloem vessels.

Does legume nitrogen fixation underpin host quality for the hemiparasitic plant Rhinanthus minor?

The high quality of leguminous hosts for the parasitic plant Rhinanthus minor (in terms of growth and fecundity), compared with forbs (non-leguminous dicots) has long been assumed to be a function of the legume's ability to fix atmospheric nitrogen (N) from the air and the potential for direct transfer of compatible amino compounds to the parasite. Using associations between Rhinanthus minor and Vicia faba (Fabaceae) that receive N either exclusively via symbiotic associations with rhizobia supplying organic N fixed from N(2) or exclusively through the supply of inorganic nitrate to the substrate, the underlying reasons for the quality of legumes as hosts for this parasite are unravelled. It is shown that sole dependence of the host, V. faba, on N fixation results in lower growth of the attached parasite than when the host is grown in a substrate supplied exclusively with inorganic N. In contrast, the host plants themselves achieved a similar biomass irrespective of their N source. The physiological basis for this is investigated in terms of N and abscisic acid (ABA) partitioning, haustorial penetration, and xylem sap amino acid profiles. It is concluded that legume N fixation does not underpin the quality of legumes as hosts for Rhinanthus but rather the well-developed haustorium formed by the parasite, coupled with the lack of defensive response of the host tissues to the invading haustorium and the presence of sufficient nitrogenous compounds in the xylem sap accessible to the parasite haustoria, would appear to be the primary factors influencing host quality of the legumes.

Growth and development of the facultative root hemiparasite Rhinanthus minor after removal of its host.

Facultative plant hemiparasites exhibit optimal growth only when attached to a suitable host. After attachment, stomata of the parasite remain continuously open, thus, optimising the extraction of host xylem sap. When the host shoot was removed from the hemiparasitic Rhinanthus/barley association ~14 days after attachment, the resulting host-free attached Rhinanthus continued to grow and develop similarly well as the attached parasites. These plants, however, showed altered stomatal behaviour: their stomata were open at daytime and closed at night, whereas parasitising Rhinanthus has continuously open stomata all day and night and unattached single Rhinanthus has practically closed stomata throughout day and night. After removal of the host the root growth was strongly increased, thereby increasing the root-to-shoot ratio. Abscisic acid and cytokinin relationships became more 'normal' with the Rhinanthus roots becoming able to synthesise zeatin nucleotides and zeatin ribosides, thus, behaving much as non-parasitic plants in general. It is suggested that the degrading root system of the host plant produces signals that trigger this conversion. Two explanations for these changes are discussed, the supply of dissolved organic nitrogen by the degrading host root system and a possible strong growth of growth promoting soil microorganisms using the degrading host root system as a substrate.

Cytokinin flows from Hordeum vulgare to the hemiparasite Rhinanthus minor and the influence of infection on host and parasite cytokinins relations.

Using the facultative root hemiparasite Rhinanthus minor L. and Hordeum vulgare L. as a host, the flows, depositions and metabolism of zeatin-type cytokinins [zeatin (Z), zeatin riboside (ZR), zeatin nucleotide (ZN)] within the host, the parasite and between host and parasite have been studied during the period 41-54 d after planting (i.e. ~30-43 d after successful attachment of the parasite to the host). Parasitism decreased the synthesis of Z in the root (by 57%) and decreased xylem flows (by 56%) and metabolism (by 71%) in leaf laminae. However, phloem flows of Z were increased by 3-fold in the host barley. The deposition of Z in the roots of Rhinanthus and the flows in xylem and phloem were increased by 20, 12, 29-fold, respectively, after successfully attaching to the host barley. However, net biosynthesis of Z in Rhinanthus roots decreased by 35% after attachment. This indicates that a large portion (70%) of xylem flow of Z in attached Rhinanthus was extracted from the host. In singly growing Rhinanthus plants, the balance of Z deposition in the shoot was negative (i.e. Z was metabolised and exported back to root in the phloem). Xylem flows and deposition of ZR and ZN showed comparable quantitative changes after attachment. A significant deposition of Z, ZR and ZN was detected in the haustoria of the Rhinanthus / barley association. The possible physiological functions of the large quantities of Z and ZR and ZN derived from the host barley, for the improved leaf development and the stomatal reactions of the parasitising Rhinanthus are discussed.

Abscisic acid (ABA) flows from Hordeum vulgare to the hemiparasite Rhinanthus minor and the influence of infection on host and parasite abscisic acid relations.

Using the facultative root hemiparasite Rhinanthus minor and Hordeum vulgare as a host, the flows, depositions, and metabolism of abscisic acid (ABA) within the host, within the parasite, and between host and parasite have been studied. When the plants were supplied with 5 mM NO(3)(-), there were weak or no effects of parasitism on ABA flows, biosynthesis, and ABA degradation in barley. However, ABA deposition was significantly affected in the leaf laminae (3-fold) and in the leaf sheath (2.4-fold), but not in roots. Dramatic changes in ABA flows, metabolism, and deposition on a per plant basis, however, have been observed in Rhinanthus. Biosynthesis in the roots was 12-fold higher after attachment, resulting in 14-fold higher ABA flows in the xylem. A large portion of this ABA was metabolized, a small portion was deposited. Phloem flows of ABA were increased 13-fold after attachment. The concentrations of ABA in tissues and transport fluids were higher in attached Rhinanthus by an order of magnitude than in host tissues and xylem sap. The same tendency was also found in a comparison between single Rhinanthus and unparasitized barley. As compared with 5 mM NO(3)(-), lower NO(3)(-) or 1 mM NH(4)(+) supply doubled the ABA concentrations in barley leaf laminae, while having only small or non-significant effects in the other organs. The possible function of ABA for the parasite is discussed.

Solute flows from Hordeum vulgare to the hemiparasite Rhinanthus minor and the influence of infection on host and parasite nutrient relations.

Using the facultative root hemiparasite Rhinanthus minor and Hordeum vulgare L. as a host, the flows and partitioning of mineral nutrients within the host, the parasite and between host and parasite have been studied during the study period 41-54 d after planting, i.e approximately 30-43 d after successful attachment of the parasite to the host. In parasitising Rhinanthus shoot growth was 12-fold, but root growth only 2-fold increased compared to non-parasitising plants. Conversely, in the Hordeum host, shoot dry matter growth was clearly reduced, by 33% in leaf laminae and by 52% in leaf sheaths, whereas root growth was only slightly reduced as a consequence of parasitism. Growth-dependent increments of total nitrogen (N) and phosphorus (P) and of potassium (K), calcium (Ca) and magnesium (Mg) in parasitising Rhinanthus shoot were strongly increased, particularly increments of total N and P, which were 18 and 42 times, respectively, higher than in solitary Rhinanthus. However, increments of the above mineral nutrients in leaf sheaths of parasitised Hordeum vulgare were more strongly decreased than in leaf laminae in response to parasitic attack. Estimation of the flows of nutrients revealed that Rhinanthus withdrew from the host xylem sap about the same percentage of each nutrients: 18% of total N, 22% of P and 20% of K. Within the host almost all net flows of nutrient ions were decreased due to parasitism, but retranslocation from shoot to root was somewhat increased for all nutrients. Quantitative information is provided to show that the substantially increased growth in the shoot of attached Rhinanthus and the observed decrease in Hordeum shoot growth after infection were related to strongly elevated supply of nitrogen and phosphorus in the parasite and to incipient deficiency of these nutrients in the parasitised host. The flows of nutrients between host and parasite are discussed in terms of low selectivity of nutrient abstraction from the host xylem by the hemiparasite Rhinanthus minor.

Water flows in the parasitic association Rhinanthus minor/Hordeum vulgare.

Using the facultative root hemiparasite Rhinanthus minor and its host Hordeum vulgare several aspects of water relations have been measured in this parasitic association. Extraction of xylem sap by the parasite from the host's roots is facilitated by con siderably higher transpiration per leaf area in the parasite than in the host and by the fact that stomata of attached Rhinanthus were open all day and night despite extremely high ABA concentrations in the leaves. By comparison, another root hemiparasite, Melampyrum arvense, parasitizing various grasses in the field, showed normal diurnal stomatal behaviour. The abnormal behaviour of Rhinanthus stomata was not due to anatomical reasons as closure could be induced by applying high external ABA concentrations. Remarkable differences have been detected between the hydraulic conductance of barley seminal roots showing relatively low values and that of Rhinanthus seminal roots showing very high values. The latter could be related to the observed high ABA concentrations in these roots. Whole plant water uptake, transpirational losses, growth-dependent deposition, and the flows of water within the plants have been measured in singly growing Rhinanthus and Hordeum plants and in the parasitic association between the two. Water uptake, deposition and transpiration in Rhinanthus were dramatically increased after attachment to the barley host; most of the water used by the parasite was extracted as xylem sap from the host, thereby scavenging 20% of the total water taken up by the host's roots. This water uptake by the parasitized host, however, due to a parasite-induced reduction in the host's growth, was decreased by 22% as compared to non-parasitized barley. The overall changes in growth-related water deposition in the host and parasite pointed to decreased shoot growth and relatively favoured root growth in the host and to strongly favoured shoot growth in the parasite. These changes in the host became more severe, when more than one Rhinanthus was parasitizing one barley plant.

Flows of elements, ions and abscisic acid in Ricinus communis and site of nitrate reduction under potassium limitation.

In a pot experiment Ricinus communis plants were cultivated in quartz sand and supplied daily with a nutrient solution which contained 4 mol m(-3) nitrate as the nitrogen source and either full strength potassium (1.3 mol m(-3), control) or 8% potassium (0.1 mol m(-3), K(+)-limitation). Although the final fresh weight of the whole plant was not affected by K(+)-limitation, the root-shoot ratio was increased due to a relatively increased root growth and inhibited development of younger shoot parts. Owing to K(+)-limitation, photosynthesis was slightly decreased, while dark respiration of the shoot markedly decreased and root respiration was nearly doubled. The transport of carbon in the phloem, and to some extent in the xylem, was greater and the root was favoured in the partitioning of carbon. This was also true for nitrogen and potassium which were both taken up at lower rates, particularly potassium. In these two cases a high remobilization and recycling from the old part of the shoot was observed. By contrast, uptake of sodium was 2.4-fold higher under K(+)-limitation and this resulted in increased flows in the plants, which was discussed generally as a means for charge balance (in combination with a slight increase in uptake of magnesium and calcium). Nitrate reduction took place in the same portion in the root and shoot. This was a shift to the root compared to the control and points to an inhibition of xylem transport caused by limitation of K(+) as an easily permeating countercation. Low K(+) supply also resulted in an increased biosynthesis of ABA in the roots (265%). This caused a slightly increased deposition of ABA in the roots (193%) and a 4.6-fold higher root-to-shoot and a doubled shoot-to-root ABA signal in the xylem or phloem, respectively. The high degradation of ABA in the shoots prevented ABA accumulation there.

Foliar application of nitrate or ammonium as sole nitrogen supply in Ricinus communis. II. The flows of cations, chloride and abscisic acid.

Following a precultivation with pedospheric nitrogen nutrition, Ricinus plants were supplied with nitrogen solely by spraying nitrate or ammonium solution onto the leaves during the experimental period. The chemical composition of tissues, xylem and phloem exudates was determined and on the basis of the previously determined nitrogen flows (Peuke et al., New Phytologist (1998), 138, 657-687) the flows of potassium, sodium, magnesium, calcium, chloride and ABA were modelled. These data, which permit quantification of net-uptake, transport in xylem and phloem, and utilization in shoot and root, were compared with results obtained in plants with pedospherically-supplied nitrate or ammonium and data in the literature. Although the overall effects on the chemical composition of supplying ammonium to the leaves were not as pronounced as in pedospherically supplied plants, there were some typical responses of plants fed with ammonium (ammonium syndrome). In particular, in ammonium-sprayed plants uptake and transport of magnesium decreased and chloride uptake was increased compared with nitrate-sprayed plants. Furthermore, acropetal ABA transport in the xylem in ammonium-sprayed Ricinus was threefold higher than in nitrate-sprayed plants. Additionally, concentrations of anions were more or less increased in tissues, particularly in the roots, and transport fluids. The overall signal from ammonium-sprayed leaves without a direct effect of ammonium ions on uptake and transport systems in the root is discussed.

Ion distribution in relation to leaf age in Leptochloa fusca* (L.) Kunth (Kallar grass): I. K, Na, Ca and Mg.

Using Leptochloa fusca (Kallar grass) plants, the distribution of K, Na, Mg and Ca between leaves of various ages has been studied. Plants grown in a salt-affected, reclaimed field, in a solution culture and in soil in pots at 10, 100 and 125 mM NaCl have been analyzed. Despite the presence of salt-secreting glands on Leptochloa fusca leaves, Na concentrations increased strongly with leaf age, while K concentrations were highest in young leaves and decreased with increasing leaf age. This was due to K retranslocation, which was clearly intensified at higher external NaCl concentration. The data point to the importance of K recycling in this halophyte and also to the use of Na by this salt-secreting species for turgor maintenance in mature and old leaves. Both Mg and particularly Ca concentrations increased with leaf age, indicating that xylem import surpassed phloem export even for the phloem-mobile ion, Mg. In young leaves, Mg concentrations exceeded those of Ca.

Ion distribution in relation to leaf age in Leptochloa fusca (L.) Kunth* (Kallar grass) II. Anions*†.

Using Leptochloa fusca (L.) Kunth (Kallar grass) plants, the distribution of Cl- , NO3 - , H3 PO4 - , SO4 2- and malate between leaves of various ages has been studied. Plants grown in a reclaimed, salt-affected field, in solution culture and in soil at 10, 100 and 125 mM NaCl have been analyzed. Apparently due to excretion by salt secreting glands on L. fusca leaves and to phloem export, Cl- concentrations did not increase strongly with leaf age. On a leaf f. wt basis, chloride secretion was constant over the series of increasingly aged mature leaves. If it was related to the chloride increments in the leaves, chloride secretion increased strongly from younger to mature leaves and reached between ISO and 200% of the concurrent Cl- deposition in the lamina. Changes in the tissue concentrations of nitrate and phosphate with leafage showed a maximum in recently matured leaves. Decreases in older leaves were attributed to nitrate reduction and export of reduced nitrogen and to retranslocation of phosphate. In leaves of field-grown L. fusca nitrate was non-detectable. Sulphate and malate concentrations in laminae continued to increase from the youngest to the oldest leaves. The increasing negative charge resulting from these increases in divalent anions can be accounted for by the loss of charge occurring in connection with reduction of nitrate and export of phosphate. Higher external salinity led, apart from increases in tissue Cl- , to noticeable decreases in tissue nitrate and phosphate but not in sulphate and malate concentrations, the latter being even increased at higher external NaCl. The observed changes in anion concentrations are compared with and discussed in relation to changes found in Ricinus communis and in Atriplex hortensis.