Comparisons were made between the free amino acid composition in leaf exudates and that in pure phloem sap, using twin samples taken from a single leaf of two oat (Avena sativa L.) and three barley (Hordeum vulgare L.) varieties. Leaf exudate was collected in a 5 mm EDTA-solution (pH 7.0) from cut leaf blades and phloem sap was obtained through excised aphid (Rhopalosiphum padi L.) stylets. Fluorescent derivatives of amino acids were obtained using 9-fluorenylmethyl chloroformate and were separated by means of high performance liquid chromatography. The total concentration of free amino acids varied considerably in the exudate samples. There was no correlation between the total amino acid content in the exudate samples and that of the corresponding phloem sap samples, but the amino acid composition of the corresponding samples was highly correlated (median R2-value 0.848). There was only limited between-plant variation in phloem sap amino acid composition. Nevertheless, in comparisons involving all samples, many of the amino acids showed significant correlations between their relative amounts in exudate and phloem sap. The results presented here indicate that the exudate technique holds great promise as an interesting alternative to the laborious and time-consuming stylet-cutting technique of obtaining samples for comparative studies of phloem sap.parts of large aphids (4) but as most aphids are small, mechanical excision of their stylets is very difficult. Despite recent technological advances, including the adoption of high frequency microcautery (9) and ruby lasers (1, 3) for cutting the mouthparts, the aphid stylet technique remains laborious and time-consuming and, usually, only small quantities of phloem sap are obtained.The exudation technique involves collection of the sap exuding from cut plant parts. King and Zeevaart (5)
High performance liquid chromatography was used for amino acid analysis of phloem sap from oats and barley. The sap was obtained from severed stylets of the aphid Rhopalosiphum padi (L.) excised by means of high frequency microcautery. Qualitative differences between the cereals were negligible. The proportions of different amino acids largely resembled those demonstrated for whole leaf extracts. Samples gave very weak detector responses when dissolved in water and kept at room temperature. Explanations for this are discussed and a technique to prevent this deterioration is described.
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