Biochemical study of leaf browning in minimally processed leaves of lettuce (Lactuca sativa L. var. acephala).

A series of biochemical parameters, including the concentration of total ascorbic acid (ASA(tot)) and the activities of phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), and peroxidases (PODs), was investigated during cold storage (72 h at 4 degrees C in the dark) in fresh-cut (minimally processed) leaves of two lettuce (Lactuca sativa L. var. acephala) cultivars differing in the susceptibility to tissue browning: Green Salade Bowl (GSB), susceptible, and Red Salade Bowl (RSB), resistant. The two cultivars showed differences also at the biochemical level. The content in ASA(tot) increased in RSB, as a consequence of increased DHA concentration; conversely, ASA(tot) diminished in GSB, in which ASA was not detectable after 72 h of storage, thus suggesting a disappearance of ascorbate (both ASA and DHA) into nonactive forms. The antioxidant capacity (as determined by using FRAP analysis) decreased significantly during storage in RSB, while a strong increase was observed in GSB. PAL activity increased soon after processing reaching a maximum by 3 h, then it declined to a relatively constant value in RSB, while in GSB it showed a tendency to decrease in the first few hours from harvest and processing. POD activity, at least for chlorogenic acid, increased significantly during storage only in GSB.

How the roots contribute to the ability of Phaseolus vulgaris L. to cope with chilling-induced water stress.

Intact plants and stem-girdled plants of Phaseolus vulgaris grown hydroponically were exposed to 5 degrees C for up to 4 d; stem girdling was used to inhibit the phloem transport from the leaves to the roots. After initial water stress, stomatal closure and an amelioration of root water transport properties allowed the plants to rehydrate and regain turgor. Chilling augmented the concentration of abscisic acid (ABA) content in leaves, roots and xylem sap. In intact plants stomatal closure and leaf ABA accumulation were preceded by a slight alkalinization of xylem sap, but they occurred earlier than any increase in xylem ABA concentration could be detected. Stem girdling did not affect the influence of chilling on plant water relations and leaf ABA content, but it reduced slightly the alkalinization of xylem sap and, principally, prevented the massive ABA accumulation in root tissues and the associated transport in the xylem that was observed in non-girdled plants. When the plants were defoliated just prior to chilling or after 10 h at 5 degrees C, root and xylem sap ABA concentration remained unchanged throughout the whole stress period. When the plants were chilled under conditions preventing the occurrence of leaf water deficit (i.e. at 100% relative humidity), there were no significant variations in endogenous ABA levels. The increase in root hydraulic conductance in chilled plants was a response neither to root ABA accretion, nor to some leaf-borne chemical signal transported downwards in the phloem, nor to low temperature per se, as indicated by the results of the experiments with defoliated or girdled plants and with plants chilled at 100% relative humidity. It was concluded that the root system contributed substantially to the bean's ability to cope with chilling-induced water stress, but not in an ABA-dependent manner.

Involvement of Abscisic Acid in Regulating Water Status in Phaseolus vulgaris L. during Chilling.

During the first hours of chilling, bean (Phaseolus vulgaris L., cv Mondragone) seedlings suffer severe water stress and wilt without any significant increase in leaf abscisic acid (ABA) content (P. Vernieri, A. Pardossi, F. Tognoni [1991] Aust J Plant Physiol 18: 25-35). Plants regain turgor after 30 to 40 h. We hypothesized that inability to rapidly synthesize ABA at low temperatures contributes to chilling-induced water stress and that turgor recovery after 30 to 40 h is mediated by changes in endogenous ABA content. Entire bean seedlings were subjected to long-term (up to 6 d) chilling (3 degrees C, 0.2-0.4 kPa vapor pressure deficit, 100 mumol.m(-2).s(-1) photosynthetic photon flux density, continuous fluorescent light). During the first 24 h, stomata remained open, and plants rapidly wilted as leaf transpiration exceeded root water absorption. During this phase, ABA did not accumulate in leaves or in roots. After 24 h, ABA content increased in both tissues, leaf diffusion resistance increased, and plants rehydrated and regained turgor. No osmotic adjustment was associated with turgor recovery. Following turgor recovery, stomata remained closed, and ABA levels in both roots and leaves were elevated compared with controls. The application of ABA (0.1 mm) to the root system of the plants throughout exposure to 3 degrees C prevented the chilling-induced water stress. Excised leaves fed 0.1 mm ABA via the transpiration stream had greater leaf diffusion resistance at 20 and 3 degrees C compared with non-ABA fed controls, but the amount of ABA needed to elicit a given degree of stomatal closure was higher at 3 degrees C compared with 20 degrees C. These findings suggest that endogenous ABA may play a role in ameliorating plant water status during chilling.

Immunological detection of acetaldehyde-protein adducts in ethanol-treated carrot cells.

Polyclonal antibodies able to recognize protein-acetaldehyde conjugates were produced and characterized. The antibodies react with sodium cyanoborohydride-reduced Schiff's bases between acetaldehyde and a protein, independently of the nature of the macromolecule binding the acetaldehyde moiety. Only conjugates between acetaldehyde or propionaldehyde and a protein are recognized; conjugates obtained with other aldehydes are not reactive. Results concerning the formation of acetaldehyde adducts with carrot (Daucus carota L.) proteins are presented as well as the presence of such conjugates in ethanol-treated carrot cell cultures, a system highly sensitive to the presence of ethanol in the culture medium.

Growth regulator levels in embryo and suspensor of Phaseolus coccineus at two stages of development.

Gibberellins and auxins were extracted from embryos and suspensors of Phaseolus coccineus L. at two stages of development: A) heart-shaped embryo and B) cotyledonary embryo with suspensor in the initial stage of degeneration. The time interval between the two stages was 5-6 days.In both embryos and suspensors, gibberellin (GA)-like activity was found in three fractions: F-1 (ethyl acetate fraction at pH 8.0), F-2 (free GAs) and F-3 (bound GAs). At stage A, the total GA activity in the suspensor was about 30 times greater than in the embryo and the bound GAs contributed by about 90% to the total GA content. A dramatic decrease in level of bound GA-like substances was found in suspensors at stage B, when the level of total GAs in the embryo had increased to 10 times that at stage A. This might suggest a transport of GAs from the suspensor to the embryo. In both embryo and suspensor, qualitative changes in GAs with shift in activity of the fractions tested occurred at the two developmental stages.The methanolic extracts of stage A suspensors showed two inhibitors, one much more active than the other, and two large peaks of growth promoting activity at Rf 0.4-0.7; in stage A embryos, the general activity of the extracts was lower and the promoting effect was spread over Rf 0.3-0.9.The present results seem to support the view that the suspensor plays a role in embryogenesis by acting as a site of synthesis of growth regulators needed by the embryo.

Growth of bean and tomato plants as affected by root absorbed growth substances and atmospheric carbon dioxide.

Bean and tomato plants were grown in solution culture root media containing pre-determined concentrations of gibberellin A3 (GA), 1-naphthaleneacetic acid (NAA), N(6)-benzyladenine (BA), (2-chloroethyl)trimethylammonium chloride (CCC), and at atmospheric levels of 300 and 1000 ppm of CO2. Net assimilation rates (NAR), relative growth rates (RGR), leaf area ratios (LAR), root to top dry weight ratios (R/T) and changes in dry weight, size, and form of each organ were recorded.Gibberellin had no effect on RGR of either plant species but increased the NAR of tomatoes at 1000 ppm CO2. Total dry weight was only slightly affected by GA but root growth and R/T were markedly depressed. CCC had no effect on NAR, but decreased RCR and LAR. Root growth of beans and R/T in both plants were promoted by CCC. NAR and RGR were strongly inhibited by BA and NAA. Inhibition of stem and leaf growth by CCC and NAA was greater than that for roots; thus, R/T ratios were increased. Root branching was promoted by NAA.High (1000 ppm), compared to the low (300 ppm), atmospheric levels of CO2 generally promoted root growth and produced an increase in the R/T, both in the absence and presence of chemical treatment. The multiplicity of effects of the rootabsorbed chemical growth substances and CO2 on growth and photosynthesis is discussed.