Roberto Lorenzi

Solid phase radioimmunoassay for the quantitation of abscisic acid in plant crude extracts using a new monoclonal antibody.

Summary A new mouse monoclonal antibody against abscisic acid (ABA) was produced and characterized. This antibody is highly specific for free (S)-ABA and shows a good affinity constant. A newly developed solid phase radioimmunoassay (RIA) for the quantitation of ABA content in plant extracts is described. This assay proved to be more sensitive and precise than the commonly used liquid phase RIA. Using this assay ABA was quantified in tomato and bean leaves and in Sechium endosperm. Results were compared with those obtained using the other two monoclonal antibodies produced up to now against ABA. Results obtained with RIA were validated by physicochemical methods.

Hormonal relationships in xylem sap of grafted and ungrafted Prunus rootstocks

Three Prunus species, characterised by different levels of growth vigour, were used as rootstocks for peach [Prunus persica (L.) Batsch] grafting and compared for their hormone transport rates in the sap extracted from both the grafted scions and the ungrafted rootstocks. Two of these species were hybrid rootstocks widely used in peach orchards, while the third was a commercial cultivar, which was used both as rootstock and as scion. The xylem exudates were analysed for the concentrations of indoleacetic acid and for the main cytokinins, namely zeatin, zeatin riboside, isopentenyl-adenine and isopentenyl-adenosine. In ungrafted rootstocks the growth potential was positively correlated with both indoleacetic acid and zeatin riboside transport rates. In the xylem sap of peach scions grafted on the same rootstocks, the growth potential and the zeatin riboside levels were also positively correlated, while a negative correlation was found with indoleacetic acid levels. The results are discussed in view of the most widely accepted hypothesis concerning the control exerted by rootstocks on scion growth.

Gibberellin Biosynthesis in Phaseolus coccineus Suspensor

Summary Ent(14C)-7-α-hydroxykaurenoic acid fed to a cell-free system of Phaseolus coccineus suspensors was incorporated into several products among which GA5, GA1 and GA8 were identified by GC-MS. Thus the suspensor has the enzymatic potential for the biosynthesis of the major gibberellins present in the very young whole seeds. Moreover these results clearly indicate that the suspensor is active in the biosynthesis of growth substances as well as in the transport of nutrients for the embryo (Yeung, 1980).

Indoleacetic acid concentration and metabolism changes during bud development in tubers of two potato (Solanum tuberosum) cultivars.

Plant growth regulators are involved in the control of potato (Solanum tuberosum) tuber dormancy. Evidence concerning the role of IAA is controversial; we therefore investigated its role by analyzing two cultivars with varying lengths of dormancy. We examined the time course of free and conjugated IAA in tuber tissue isolates from the final stages of tuber growth to the end of dormancy, the distribution of free IAA in tuber tissues by in situ analysis, and the biosynthesis of the hormone by feeding experiments. The time course of free IAA showed marked differences between the examined cultivars, although the concentration of the auxin generally was the highest at the early stages of tuber dormancy. Immunodetection showed a similar pattern of IAA distribution in both genotypes: in dormant buds from freshly harvested tubers, the free hormone accumulated mostly in apical meristem, leaf and lateral bud primordia, and differentiating vascular tissues underlying the apical meristem, while at the end of the storage period only axillary bud primordia from growing buds displayed appreciable auxin levels. Feeding experiments indicated that changes in IAA biosynthesis rate were a major cause of auxin variation in buds. In both cultivars, dormancy apparently ceased when free IAA fell below a threshold value. Despite this, our data led us to conclude that IAA would not be directly responsible for inhibiting sprouting. Instead, auxin might shorten dormancy, in a cultivar-dependent manner, by enhancing early developmental processes in buds, ultimately leading to dormancy termination.

Changes in free and conjugated IAA during dormancy and sprouting of potato tubers

The time course of free and conjugated indole-3-acetic acid (IAA) concentrations was investigated in three distinct parts (eyes, subeye tissues and pith) of potato (Solanum tuberosum L. cultivar Monalisa) tubers, from the last period of tuber growth to sprouting under two different storage temperatures (3 and 23˚C). The highest levels of both free and conjugated IAA were detected in eyes, which showed a significant increase of the free hormone concentration from harvest to the end of dormancy, regardless of storage temperature. The concentration of conjugates was higher than free IAA in eyes and subeye tissues at 23˚C, while at 3˚C conjugates prevailed only in eyes. Present results suggest the involvement of IAA in the control of potato tuber dormancy.

Changes in ultrastructure, protease and caspase-like activities during flower senescence in Lilium longiflorum

The last phase of flower development is senescence during which nutrients are recycled to developing tissues. The ultimate fate of petal cells is cell death. In this study we used the ethylene-insensitive Lilium longiflorum as a model system to characterize Lily flower senescence from the physiological, biochemical and ultrastructural point of view. Lily flower senescence is highly predictable: it starts three days after flower opening, before visible signs of wilting, and ends with the complete wilting of the corolla within 10 days. The earliest events in L. longiflorum senescence include a fall in fresh and dry weight, fragmentation of nuclear DNA and cellular disruption. Mesophyll cell degradation is associated with vacuole permeabilization and rupture. Protein degradation starts later, coincident with the first visible signs of tepal senescence. A fall in total protein is accompanied by a rise in total proteases, and also by a rise of three classes of caspase-like activity with activities against YVAD, DEVD and VEID. The timing of the appearance of these caspase-like activities argues against their involvement in the regulation of the early stages of senescence, but their possible role in the regulation of the final stages of senescence and cell death is discussed.

Nitric oxide and hydrogen peroxide involvement during programmed cell death of Sechium edule nucellus

The nucellus is a maternal tissue that feeds the developing embryo and the secondary endosperm. During seed development the cells of the nucellus suffer a degenerative process early after fertilization as the cellular endosperm expands and accumulates reserves. Nucellar cell degeneration has been characterized as a form of developmentally programmed cell death (PCD). In this work we show that nucellus PCD is accompanied by a considerable production of both nitric oxide and hydrogen peroxide (NO and H(2)O(2)). Interestingly, each of the two molecules is able to induce the production of the other and to cause cell death when applied to a living nucellus. We show that the induced cell death has features of a PCD, accompanied by profound changes in the morphology of the nuclei and by a massive degradation of nuclear DNA. Moreover, we report that NO and H(2)O(2) cause an induction of caspase-like proteases previously characterized in physiological nucellar PCD.

The effects of (S)-(+)-carvone treatments on seed potato tuber dormancy and sprouting

SummaryPotato seed tubers may suffer from premature sprouting during storage, thus limiting their suitability for cultivation. Commonly used sprout suppressant treatments negatively affect but viability and therefore a reliable method to inhibit bud development must still be found for seed tubers. The monoterpene carvone ((S)-(+)-carvone) was tested in small scale experiments. The vapour of this compound fully inhibited bud growth of tubers cv. Monalisa stored at 23°C without affecting bud viability throughout 6 months of treatment. The most effective range of carvone vapour concentrations was between 0.34 and 1.06 μmol mol−1. With these qualities we can expect carvone to become a suitable sprout suppressant for seed tubers.

Role and metabolism of abscisic acid in potato tuber dormancy and sprouting

Summary Tubers cv. Monalisa were analysed to determine the content of abscisic acid (c-t-(+)ABA) and related metabolites in three tuber parts (eyes, sub-eye tissues and pith). Analyses were performed from the last period of tuber growth until sprouting under two different storage temperatures (3 °C and 23 °C). ABA content rises in eyes and sub-eye tissues as sprouting approaches, regardless of storage temperature. Dihydrophaseic acid is the main ABA metabolite while phaseic acid was not detected, probably because of its further metabolization to DPA. ABA-glucose ester concentrations are generally low in all tissues except in eyes before harvest. The results are discussed in relation to the current hypotheses on the role of ABA in the control of potato tuber dormancy.

Kaurene and kaurenol biosynthesis in cell-free system of Phaseolus coccineus suspensor

Abstract It is shown that suspensor tissue of Phaseolus coccineus can biosynthesize ent -kaur-16-ene and ent -kaur-16-en-19β-ol, two key precursors in the biosynthesis of gibberellins.