Photoprotective Role of Photosynthetic and Non-Photosynthetic Pigments in Phillyrea latifolia: Is Their "Antioxidant" Function Prominent in Leaves Exposed to Severe Summer Drought?

Carotenoids and phenylpropanoids play a dual role of limiting and countering photooxidative stress. We hypothesize that their "antioxidant" function is prominent in plants exposed to summer drought, when climatic conditions exacerbate the light stress. To test this, we conducted a field study on Phillyrea latifolia, a Mediterranean evergreen shrub, carrying out daily physiological and biochemical analyses in spring and summer. We also investigated the functional role of the major phenylpropanoids in different leaf tissues. Summer leaves underwent the most severe drought stress concomitantly with a reduction in radiation use efficiency upon being exposed to intense photooxidative stress, particularly during the central hours of the day. In parallel, a significant daily variation in both carotenoids and phenylpropanoids was observed. Our data suggest that the morning-to-midday increase in zeaxanthin derived from the hydroxylation of ß-carotene to sustain non-photochemical quenching and limit lipid peroxidation in thylakoid membranes. We observed substantial spring-to-summer and morning-to-midday increases in quercetin and luteolin derivatives, mostly in the leaf mesophyll. These findings highlight their importance as antioxidants, countering the drought-induced photooxidative stress. We concluded that seasonal and daily changes in photosynthetic and non-photosynthetic pigments may allow P. latifolia leaves to avoid irreversible photodamage and to cope successfully with the Mediterranean harsh climate.

Fraxitoxin, a New Isochromanone Isolated from Diplodia fraxini.

A new isochromanone, named fraxitoxin, was isolated together with (-)-mellein and tyrosol from liquid cultures of Diplodia fraxini, a pathogen involved in the etiology of canker and dieback disease of Fraxinus spp. in Europe. It was characterized as 5-methoxy-3-methylisochroman-1-one using spectroscopic methods (essentially NMR and HR-EI-MS). Its absolute configuration (R) at C(3) was assigned by electronic circular dichroism (ECD) measurements and calculations. Phytotoxic activity of the compound was evaluated on ash, cork and holm oak leaves at concentration of 1 mg/ml by the leaf-puncture assay. Interestingly, fraxitoxin caused necrotic lesions only on ash leaves.

Assessment of factors affecting micropropagation and ex vitro acclimatization of Nyctanthes arbor-tristis L.

Rapid differentiation of multiple shoots was observed in 94% of nodal explants of one year old Nyctanthes arbor-tristis L. plants. Shoot bud induction and multiplication took place on Murashige and Skoog (MS) medium supplemented with two cytokinins, i.e. Benzyladenine (BA) or Kinetin (Kn) either alone or in combination with different auxins, indole-3-butyric acid (IBA), indole-3-acetic acid (IAA) or α-naphthalene acetic acid (NAA). Between different media, pH levels and growth regulators tried, the optimum condition for maximum regenerative response was obtained on MS + Kn (2.5 µM) + N AA (0.5 µM) media at 5.8 pH, forming cultures with 23.26 ± 0.89 number of shoots and 6.36 ± 0.80 cm shoot length after 8 weeks of culture. Histological sections confirmed the formation of multiple buds from nodal explants. Rooting was achieved ex vitro by dipping the basal ends of microshoots in 200 µM IBA for 30 min followed by their transplantation in sterile soilrite. The plantlets with well-developed shoot and root system were successfully established in garden soil and grown outside in a greenhouse with a 80% survival rate.

Preconditioning of axillary buds in thidiazuron-supplemented liquid media improves in vitro shoot multiplication in Nyctanthes arbor-tristis L.

An efficient tissue culture technology has been designed for mass multiplication of Nyctanthes arbor-tristis L. by preculturing nodal explants in thidiazuron (TDZ)-supplemented liquid Murashige and Skoog (MS) media. Direct inoculation of nodal segments on semi-solid MS medium augmented with various concentrations of TDZ (0.1 to 0.9 µM) produced shoots but with low regeneration response and few shoots per explant. Hence, nodal explants were pretreated with greater concentrations of TDZ (5 to 100 µM) in liquid MS media for different durations (4, 8, 12, and 16 days) with the aim of improving shoot regeneration response from cultured explants. After pretreatment, explants were transferred to agar-solidified hormone-free MS medium. Best response in terms of percent regeneration (94%), number of shoots per explant (20.00 ± 1.15), and greatest shoot length (7.23 ± 0.83 cm) were obtained with nodal segments pretreated in 75 µM TDZ for 8 days. Similarly, root induction was obtained from pulse-treated microshoots for 24 h with 200 µM indole-3-butyric acid (IBA) followed by their transfer to 1/2 MS medium which produced an average of 5.50 ± 0.92 roots per microshoot. The rooted plantlets were transplanted to soil with 80% success rate.

Responses to changes in Ca2+ supply in two Mediterranean evergreens, Phillyrea latifolia and Pistacia lentiscus, during salinity stress and subsequent relief.

BACKGROUND AND AIMS: Changes in root-zone Ca(2+) concentration affect a plant's performance under high salinity, an issue poorly investigated for Mediterranean xerophytes, which may suffer from transient root-zone salinity stress in calcareous soils. It was hypothesized that high-Ca(2+) supply may affect differentially the response to salinity stress of species differing in their strategy of Na(+) allocation at organ level. Phillyrea latifolia and Pistacia lentiscus, which have been reported to greatly differ for Na(+) uptake and transport rates to the leaves, were studied. Methods In plants exposed to 0 mM or 200 mM NaCl and supplied with 2.0 mM or 8.0 mM Ca(2+), under 100 % solar irradiance, measurements were conducted of (a) gas exchange, PSII photochemistry and plant growth; (b) water and ionic relations; (c) the activity of superoxide dismutase and the lipid peroxidation; and (d) the concentration of individual polyphenols. Gas exchange and plant growth were also estimated during a period of relief from salinity stress. Key Results The performance of Pistacia lentiscus decreased to a significantly smaller degree than that of Phillyrea latifolia because of high salinity. Ameliorative effects of high-Ca(2+) supply were more evident in Phillyrea latifolia than in Pistacia lentiscus. High-Ca(2+) reduced steeply the Na(+) transport to the leaves in salt-treated Phillyrea latifolia, and allowed a faster recovery of gas exchange and growth rates as compared with low-Ca(2+) plants, during the period of relief from salinity. Salt-induced biochemical adjustments, mostly devoted to counter salt-induced oxidative damage, were greater in Phillyrea latifolia than in Pistacia lentiscus. CONCLUSIONS: An increased Ca(2+) : Na(+) ratio may be of greater benefit for Phillyrea latifolia than for Pistacia lentiscus, as in the former, adaptive mechanisms to high root-zone salinity are primarily devoted to restrict the accumulation of potentially toxic ions in sensitive shoot organs.

Gene expression induced by chronic ozone in the Mediterranean shrub Phillyrea latifolia: analysis by cDNA-AFLP.

Seedlings of Phillyrea latifolia L., a Mediterranean shrub, were exposed for 90 days to 110 nl l(-1) ozone (O(3)). Comparison of the cDNA-amplified fragment length polymorphism (cDNA-AFLP) patterns for treated and control plants allowed the identification and cloning of 88 differential sequences induced by O(3). The differential expression of 67 cloned sequences was further confirmed by RT-PCR. The functions of 36 cloned sequences, corresponding to seven of the twelve gene functional classes of Arabidopsis, were presumed on the basis of their homology with characterized gene sequences. Ozone induction of genes homologous to 24 of the clones has been reported in other plant species, whereas the induction of the 12 remaining sequences has not been observed before. Ozone activation of these newly identified genes could be a result of the chronic exposure to low O(3) concentration, because in most previous studies, acute treatments, involving high O(3) dosages, were applied. Possible roles of the cloned sequences in the response of P. latifolia to O(3) and other causes of oxidative stress are discussed.