Effect of heat shock on the chilling sensitivity of trichomes and petioles of African violet (Saintpaulia ionantha).

Chilling at 6 degrees C caused an immediate cessation of protoplasmic streaming in trichomes from African violets (Saintpaulia ionantha), and a slower aggregation of chloroplasts in the cells. Streaming slowly recovered upon warming to 20 degrees C, reaching fairly stable rates after 4, 15, 25 and 35 min for tissue chilled for 2 min and for 2, 14 and 24 h, respectively. The rate of ion leakage from excised petioles into an isotonic 0.2 M mannitol solution increased after 12 h of chilling and reached a maximum after 3 days of chilling. A heat shock at 45 degrees C for 6 min reduced chilling-induced rates of ion leakage from excised 1-cm petiole segments by over 50%, namely to levels near that from non-chilled control tissue. Heat-shock treatments themselves had no effect on the rate of ion leakage from non-chilled petiole segments. Protoplasmic streaming was stopped by 1 min of heat shock at 45 degrees C, but slowly recovered to normal levels after about 30 min Chloroplasts aggregation was prevented by a 1 or 2 min 45 degrees C heat-shock treatment administered 1.5 h before chilling, but heat-shock treatments up to 6 min only slightly delayed the reduction in protoplasmic streaming caused by chilling. Tradescantia virginiana did not exhibit symptoms associated with chilling injury in sensitive species (i.e. cessation of protoplasmic streaming in stamen hairs and increased ion leakage from leaf tissue).

Effect of acetaldehyde, arsenite, ethanol, and heat shock on protein synthesis and chilling sensitivity of cucumber radicles.

Exposure to a chilling temperature of 2.5 degrees C for 96 h inhibited the subsequent growth of cucumber seedling radicles at 25 degrees C by 92%. Exposing seedling with 5 +/- 1 mm long radicles to acetaldehyde vapour (275 micro l l(-1)) or to an aqueous ethanol solution (0.6 M) for 2 h, or to 45 degrees C for 10 min before chilling, increased chilling tolerance so that the chilling treatment reduced growth by only 47, 39 or 36%, respectively. All of these effective treatments induced the synthesis of a number of proteins, and suppressed de novo protein synthesis (i.e. the incorporation of [(35)S]-methionine) by about 70%. In contrast, treatment for 2 h with an aqueous arsenite solution (100 micro M) had no effect on chilling sensitivity or the incorporation of [(35)S]-methionine, yet it induced the synthesis of a complement of proteins that were similar to that induced by the effective heat-shock treatment. A unique protein or set of proteins may be responsible for heat-shock-induced chilling tolerance, but none was detected. The ability of various abiotic stresses to suppress protein synthesis may be more important in increasing tolerance to chilling injury than their ability to induce the synthesis of specific proteins.

Antioxidant capacity of lettuce leaf tissue increases after wounding.

Wounding induced the accumulation of phenolic compounds in Iceberg and Romaine lettuce leaf tissue. Phenolic concentrations were quantified after holding the leaf tissue at 10 degrees C for 48 h as the absorbance of a methanol extract at 320 nm, and by the Folin-Ciocalteu method. Heat-shock treatments applied by immersing tissue in 45 degrees C water for 2.5 min before or after wounding reduced the accumulation of phenolic compounds. Compared to the nonwounded, nonheat-shocked controls, these and other wounding and heat-shock treatments produced leaf tissue with a 4-fold range in phenolic content. The antioxidant capacity of the tissue, measured as DPPH (alpha,alpha-diphenyl-beta-picrylhydrazyl)-radical scavenging activity, or as ferric-reducing antioxidant power (FRAP), increased after wounding. The increase was linearly correlated with the increase in phenolic compounds in Iceberg (R(2) > 0.97) and in Romaine (R(2) > 0.95) lettuce leaf tissue. Increased consumption of diets rich in phenolic antioxidants may contribute to reducing human diseases. Treatments that reduce the browning of wounded lettuce leaf tissue by preventing the oxidation of the accumulated wound-induced phenolic compounds may produce a healthier fresh-cut product than treatments that prevent the wound-induced synthesis and accumulation of phenolic compounds with antioxidant properties.

Chilling tolerance of maize, cucumber and rice seedling leaves and roots are differentially affected by salicylic acid.

Salicylic acid (SA) is one component of a complex signalling pathway that is induced by a number of biotic and abiotic stresses. Exposing seedling radicles to aqueous solutions of 0.5 mM salicylic acid for 24 h before chilling at 2.5 degrees C for 1-4 days reduced the chilling-induced increase in electrolyte leakage from maize and rice leaves, and cucumber hypocotyls, but not from their radicles. The SA treatments that induced chilling tolerance in the aerial portion of the seedlings did not induce chilling tolerance in the radicles, even though the SA treatments were applied to the radicles. A comparison of activity among five antioxidant enzymes showed that SA did not alter enzyme activities in the radicles, but that chilling tolerance induced by SA in the aerial portions of maize and cucumber plants was associated with an increase in the activity of glutathione reductase and guaiacol peroxidase.

Reduced chilling tolerance in elongating cucumber seedling radicles is related to their reduced antioxidant enzyme and DPPH-radical scavenging activity.

Cucumber seedling radicles become more chilling sensitive as they elongate. Chilling seedlings with radicles 20 mm long for 48 h at 2.5 degrees C inhibited subsequent growth by 36%, while it reduced the growth of 70 mm-long radicles by 63%. Although the growth rate of non-chilled cucumber radicles at 25 degrees C is constant from 20 to 80 mm, tissue viability [i.e. reduction of TTC (2,3,5-triphenyltetrazolium chloride) to formazan] and DPPH (alpha,alpha-diphenyl-beta-picrylhydrazyl) radical scavenging activity of apical tissue declines as radicles elongate from 20 to 80 mm in length. TTC reduction, DPPH-radical scavenging activity and protein content of apical tissue were higher in 20 than in 70 mm radicles immediately after chilling and after an additional 48 h of growth at 25 degrees C. Catalase (CAT; EC 1.11.1.6) and ascorbate peroxidase (APX; EC 1.11.1.11) activity was higher in the apical tissue of 20 than in 70 mm radicles before chilling. Immediately after chilling and after an additional 48 h at 25 degrees C, superoxide dismutase (SOD; EC 1.15.1.1), glutathione reductase (GR; EC 1.6.4.2), and guaiacol peroxidase (GPX; EC 1.11.1.7) activity increased more rapidly in 70 mm radicles than in 20 mm radicles (SOD, GR, and GPX activity in 70 mm radicles was 1.5-, 1.9- and 8.6-fold higher, respectively, than in 20 mm radicles). However, APX and CAT activity in 20 mm radicles were always higher than in 70 mm radicles. Growth after chilling enhanced the activity of all antioxidant enzymes compared to that found in non-chilled tissue; however, CAT activity in 70 mm radicles did not recover to levels found in non-chilled tissue. Higher levels of CAT, APX and DPPH-radical scavenging activity are correlated with higher chilling tolerance of 20 mm-long cucumber radicles compared to 70 mm-long radicles.

Involvement of putative chemical wound signals in the induction of phenolic metabolism in wounded lettuce.

Cutting leaves of Romaine lettuce (Lactuca sativa L. cv. Longifolia) produces a wound signal that induces the synthesis of phenylalanine ammonia lyase (PAL, EC 4.3.1.5) and the accumulation of phenolic compounds in cells up to 2 cm from the site of injury, and tissue browning near the site of injury. The response of leaves within a head of Romaine lettuce to putative chemical wound signals [abscisic acid (ABA), jasmonate (JA) and methyl jasmonate (MeJA)] differed significantly with leaf age. Exposure of harvested heads of lettuce to ABA, JA, MeJA, or salicylic acid (SA) did not induce changes in PAL activity, the concentration of phenolic compounds or browning in mature leaf tissue that was similar to the level induced by wounding. Methyl jasmonate applied as vapour (10, 100 or 1000 &mgr;l kg-1 FW), or as an aqueous spray or dip (0.01-100 &mgr;M) at 5 or 10 degrees C did not produce an effect on PAL activity or browning that differed significantly from the untreated controls. In contrast, JA, MeJA and SA did induce elevated levels of PAL activity in younger leaves. However, the levels induced were far lower than those induced by wounding. Wound induced phenolic metabolism in mature leaves appears to be induced by different signals than those functioning in young leaves.

Heat shocks increase the chilling tolerance of rice (Oryza sativa) seedling radicles.

The growth of rice (Oryza sativa L., cv. M202) seedling radicles, initially 10 +/- 1 mm long, was linear for the 96 h it took them to grow to 150 mm at 25 degrees C. Exposure to 5 degrees C for 24 h reduced the rate of growth by about 50%, and longer exposures caused a progressive reduction in growth. Initial radicle length significantly affected chilling sensitivity: with 2 days at 5 degrees C inhibiting growth at 25 degrees C by 23% for 1-mm radicles, 63% for 10-mm radicles, and 87% for 40-mm radicles. Heat shocks of 35 degrees C for 4 min, 40 degrees C for 3 min, 45 degrees C for 2 min, or 50 degrees C for 1 min, prior to chilling, reduced the 75% inhibition of radicle growth caused by 2 days at 5 degrees C to 34%, 25%, 14%, and 13%, respectively. The length of exposure that conferred chilling tolerance increased from less than 2 min for 50 degrees C to over 8 min for 35 degrees C. This increase in effective treatment duration was accompanied by a reduction in the maximum induced chilling tolerance. Practical application of heat-shock treatments to increase the chilling tolerance of rice seedlings may sacrifice a small reduction in maximum chilling tolerance at the lower inductive temperatures for a larger margin of safety in their application.

Antioxidant enzymes and DPPH-radical scavenging activity in chilled and heat-shocked rice (Oryza sativa L.) seedlings radicles.

Chilling whole rice seedlings at 5 degrees C significantly increased the time needed to recover linear growth and reduced the subsequent linear rate of radicle growth. Subjecting nonchilled seedlings to a 45 degrees C heat shock for up to 20 min did not alter subsequent growth, whereas a 3 min heat shock was optimal in reducing growth inhibition caused by 2 days of chilling. The activity of five antioxidant enzymes [superoxide dismutase (EC 1.15.1.1), catalase (CAT; EC 1.11.1.6), ascorbate peroxidase (APX; EC 1.11.1.11), glutathione reductase (GR; EC 1.6.4.2), and guaiacol peroxidase (GPX; EC 1.11.1.7)] and DPPH (1,1-diphenyl-2-picrylhydrazyl)-radical scavenging activity were measured in heat-shocked and/or chilled radicles. Heat shock slightly increased the activity of CAT, APX, and GR and suppressed the increase of GR and GPX activity during recovery from chilling. Increased CAT, APX, GR, and DPPH-radical scavenging activity and protection of CAT activity during chilling appear to be correlated with heat shock-induced chilling tolerance.