Epinasty promoted by salinity or ethylene is an indicator of salt-sensitivity in tomatoes

Abstract Six genotypes of tomato (Lycopersicon esculentum Mill.) that differ in their salt-tolerance, were exposed to 200 mol m^?3 NaCl for 4 weeks. Seedlings exhibited a marked decline in shoot dry weight accumulation and increased petiolar epinasty after exposure to salinity stress. Ranking accessions on the basis of their relative growth reduction in response to salinity, provided good agreement with the level of epinasty promoted during the salinity treatment. In the absence of salt-stress, leaf epinasty promoted by exogenous ethylene treatment was found to be a positive indicator of the genotypes incipient salt-sensitivity. Endogenous ethylene levels in untreated plants were negatively correlated with ethephon-induced epinasty. Genotypes with normally high endogenous C₂H₄ levels were less responsive to ethephon treatment and also exhibited greater salt-tolerance than genotypes with low endogenous C₂H₄ levels. These observations are consistent with the suggestion that a main feature of adaptation in the genotypes examined may involve modulation of their cellular sensitivity to C₂H₄. The results indicate that leaf epinasty, whether salt- or ethylene-induced, is a sensitive indicator of salt-sensitivity. Ethylene-induced epinasty may, therefore, provide a simple basis upon which to identify and select salt-tolerant plants.     

Prevention of salt-induced epinasty by α-aminooxyacetic acid and cobalt

Ethylene production in leaf petiole and laminae tissues was stimulated in tomato (Lycopersicon esculentum Mill. cv. UCT5) plants exposed to salinity-stress. At the highest salinity level (250 mM NaCl), rates of ethylene production more than doubled over those observed in non-stressed plants. Correspondingly, petiolar epinasty increased with increasing levels of stress impositions. Both responses were suppressed when either 1 mM -aminooxyacetic acid (AOA), or 100 M Co²⁺ was simultaneously applied. Co²⁺, but not AOA, had a pronounced effect on ethylene production resulting from the application of a saturating dose (2 mM) of 1-aminocyclopropane-1-carboxylic acid (ACC), the immediate precursor of ethylene. This result suggests that ethylene production is dependent upon the activity of ethylene forming enzyme (EFE). The magnitude of ethylene stimulation in leaf petioles was related to the salinity level imposed and to the induction of petiole epinasty. In the absence of stress impositions, epinastic responsiveness to ethylene or its precursor, ACC, might provide a simple, indirect criteria to adjudge salt-sensitivity among plants.Research supported by AID contract II, NEB-1070-A-00-2074-00.     

Growth and ionic relations of six triticale cultivars as affected by salinity

Growth and ionic relations were studied in six triticale cultivars of different geographical origins grown in a greenhouse in nutrient solution with or without the addition of 100 mM NaCl. In 21 d old plants of all the six cultivars growth was little affected in the salt treatment, whereas in the subsequent three harvests during vegetative phase (after 31, 38 and 45 d), growth reduction effects of salinity were progressively pronounced. Generally, shoots of all the six cultivars accumulated relatively more K+ as compared to Na+ or Cl-. Differential accumulation of K+, Na+ and Cl- by various cultivars was coupled with variable rates of Na+ and Cl- transport from root to shoot which were — to some extent- related to cultivar differences in growth in saline root media. Chloride content of shoots of the six cultivars was negatively correlated with the relative growth reduction due to salinity at the four harvests.     

The activity of antioxidative enzymes in three strawberry cultivars related to salt-stress tolerance

Effects of salt stress on the time course of stomatal behaviors and the activity of antioxidative enzymes such as catalase (CAT) (EC 1.11.1.6), ascorbate peroxidase (APX) (EC 1.11.1.11), and glutathione reductase (GR) (EC. 1.6.4.2) were studied in three strawberry cultivars. The responses of the cultivars ‘Camarosa’, ‘Tioga,’ and ‘Chandler’ were compared when they were irrigated with nutrient solution containing 0, 8.5, 17.0, and 34.0 mM sodium chloride (NaCl) for 30 days. A significant reduction in stomatal conductance (g_s) was seen particularly on the 30th day of the salt treatments only in Camarosa, which is parallel to transpiration rate (E). CAT activities decreased in all of the salt treatments only in Tioga, while it remained almost unchanged or slightly increased depending on the period in Camarosa and Chandler. APX activity sharply increased in 17.0 and 8.5-mM NaCl treatments for 30 days in Camarosa and Tioga, respectively, whereas it linearly increased based on the NaCl treatments in Chandler. On the other hand, only Camarosa demonstrated a sharp increase in GR activity induced by salinity applied for 30 days. All the data indicated that control of the stomatal behavior, the higher salt-stress tolerance (LT₅₀) and higher constitutive activity of antioxidant enzymes made Camarosa and Tioga relatively salt-tolerant cultivars.     

Effects of NaCl stress on proline and cation accumulation in salt sensitive and tolerant turfgrasses

Summary Concentrations of proline, sodium and potassium in shoot tissues of five turfgrass species were measured following exposure to 170 mM NaCl salinity stress. Salt tolerant ‘Fults’ alkaligrass and ‘Dawson’ red fescue restricted the accumulation of Na-ions to significatnly low levels compared to the salt sensitive Kentucky bluegrasses (‘Adelphi’ and ‘Ram I’) and ‘Jamestown’ red fescue. Accumulation of proline began in all species within 24 h of initiation of salt stress but at a more rapid rate and higher overall concentration for ‘Fults’ alkaligrass. Proline levels were variable and too low in relation to sodium accumulations to have any significant osmoregulatory role in salt tolerance among all cultivars tested with the possible exception of alkaligrass.     

Effect of salinity on phosphate accumulation and injury in soybean

Many soybean [Glycine max (L.) Merr.] genotypes that are grown in solution cultures are highly sensitive to the combination of both salinity and inorganic phosphate (Pi) in the substrate. This effect has been observed on numerous occasions on plants grown in a saline medium that contained a substantial amount of Ca (i.e., CaCl₂/NaCl=0.5 on a molar basis). Because Ca is important in regulating ion transport and membrane permeability, solution culture experiments were designed to examine the effects of various concentrations of Pi and ratios of CaCl₂/NaCl (0 to 0.5 on a molar basis) at a constant osmotic potential (−0.34 MPa) on this adverse interaction. Four soybean cultivars (‘Lee’, ‘Lee 74’ ‘Clark’ and ‘Clark 63’) were tested. No adverse salinity x Pi interaction was found on Lee at any ratio and leaf P and Cl were maintained below 300 and 200 mmol kg⁻¹ dry wt, respectively. Clark, Clark 63 and Lee 74 soybean plants, on the other hand, were severely injured by solution salinity (−0.34 MPa osmotic potential) when substrate Pi was ≥0.12 mM. Reduced substrate Ca did not intensify the salinity x Pi interaction. On the contrary, the onset of injury was hastened and more severe with increased CaCl₂/NaCl ratios in isotonic solutions. Shoot and root growth rates decreased as injury increased. Leaf P concentrations from these cultivars grown in saline solutions with 0.12 mM Pi were excessive (>600 mmol kg⁻¹ dry wt) compared with concentrations commonly found in soybean leaf tissue yet they were independent of the severity of injury. Since leaf Cl increased wiht increased CaCl₂/NaCl ratio, we suspect that the severity of foliar injury was related to the combined effects of excessive P and Cl within the tissue. Lee 74, the only injured cultivar examined that excluded Cl from its leaves, was less sensitive than either Clark cultivar and its injury was characteristically different. Other ion interactions were reported that may have played a role in injury susceptibility.     

Protein profile analysis of salt-responsive proteins in leaves and roots in two cultivars of creeping bentgrass differing in salinity tolerance

Knowledge of stress-responsive proteins is critical for further understanding the molecular mechanisms of stress tolerance. The objectives of this study were to establish a proteomic map for a perennial grass species, creeping bentgrass (A. stolonifera L.), and to identify differentially expressed, salt-responsive proteins in two cultivars differing in salinity tolerance. Plants of two cultivars (‘Penncross’ and ‘Penn-A4’) were irrigated daily with water (control) or NaCl solution to induce salinity stress in a growth chamber. Salinity stress was obtained by adding NaCl solution of 2, 4, 6, and 8 dS m⁻¹ in the soil daily for 2-day intervals at each concentration, and then by watering soil with 10 dS m⁻¹ solution daily for 28 days. For proteomic map, using two-dimensional electrophoresis (2-DE), approximately 420 and 300 protein spots were detected in leaves and roots, respectively. A total of 148 leaf protein spots and 40 root protein spots were excised from the 2-DE gels and subjected to mass spectrometry analysis. In total, 106 leaf protein spots and 24 root protein spots were successfully identified. Leaves had more salt-responsive proteins than roots in both cultivars. The superior salt tolerance in ‘Penn-A4’, indicated by shoot extension rate, relative water content, and cell membrane stability during the 28-day salinity stress could be mainly associated with its higher level of vacuolar H⁺-ATPase in roots and UDP-sulfoquinovose synthase, methionine synthase, and glucan exohydrolase in leaves, as well as increased accumulation of catalase and glutathione S-transferase in leaves. Our results suggest that salinity tolerance in creeping bentgrass could be in part controlled by an alteration of ion transport through vacuolar H⁺-ATPase in roots, maintenance of the functionality and integrity of thylakoid membranes, sustained polyamine biosynthesis, and by the activation of cell wall loosening proteins and antioxidant defense mechanisms.     

Extreme resistance to potato virus V in clones of Solanum tuberosum that are also resistant to potato viruses Y and A: evidence for a locus conferring broad-spectrum potyvirus resistance

 Extreme resistance to the potato V potyvirus (PVV) was found in four potato cultivars that contain Ry genes from Solanum stoloniferum. When plants of these cultivars, were inoculated by grafting in shoot tips from PVV-infected tomato plants, necrotic symptoms developed in some cultivars, although a full hypersensitive reaction was not elicited, while other cultivars were symptomless. PVV replication was not detected in any of the inoculated plants by ELISA, an infectivity assay of leaf extracts by manual inoculation to Nicotiana benthamiana indicator plants, or by ‘return grafting’ of shoot tips taken from newly developed shoots of the potato plants to virus-free indicator plants of tomato. These methods readily detected PVV infection in inoculated plants of cv ‘Flourball’, which does not contain an Ry gene and is susceptible, and in cvs ‘Maris Piper’ and ‘Dr Macintosh’, which contain gene Nv conditioning a hypersensitive reaction to inoculation. One of the Ry-containing cultivars, ‘Barbara’, has been previously shown to contain two genes that control extreme resistance, defined as no viral replication in intact plants, to the potyviruses potato viruses Y and A (PVY and PVA). These genes are: Ry _sto , which conditions resistance to PVY and PVA, and gene Ra, which conditions resistance to PVA only. It was found that in genotypes from a progeny of the cross ‘Barbara’ (Ry _sto /Ra)בFlourball’ (ry/ra), extreme resistance to PVV segregated with gene Ry _sto . It is proposed that either gene Ry _sto conditions broad-spectrum extreme resistance to the distinct potyviruses PVY, PVA, and PVV or that Ry _sto represents a family of genetically closely linked genes each controlling resistance to a specific virus. Received: 27 December 1996 / Accepted: 9 June 1997     

Germination kinetics and seed reserve mobilization in two flax (<Emphasis Type="Italic">Linum usitatissimum</Emphasis> L.) cultivars under moderate salt stress

Because of its high contents of protein, α-linolenic-rich oil, lignans, and fiber, demand is increasing for flax(Linum usitatissi-mum L.) and flax seed oil as a food source. In this comparative survey, we examined germination and the mobilization of seed storage products (lipids and soluble proteins) of 3-d-old seedlings from two flax cultivars (N 51 and H 52) challenged with moderate salinity (up to 200 mM NaCl). At the highest salt concentration, germination appeared to be cultivar-dependent, with that of ‘N 51’ being less impaired and delayed than in ’H 52’. Sodium chloride inhibited germination via osmotic and toxic effects, so that seed viability was altered, especially in ‘H 52’. At 200 mM NaCl, lipid mobilization was delayed in the earliest germination phases. This response was associated with increased proportions of linolenic acid contents in both cultivars and more linolenic acid-rich molecular species of TAGs. Irrespective of the salt level, soluble protein contents in both cultivars decreased over time, although a salt-related precocity of protein degradation occurred at 200 mM NaCl.     

Influence of Phosphorus on Gas Exchange and Plant Growth of Two Morphologically Distinct Types of Capsicum Annuum

Tolerance to phosphorus stress was studied in Capsicum annuum L. Chile ancho cv. San Luis and bell pepper cv. Jupiter plants. Plants were fertilized weekly with Long-Ashton nutrient solution (LANS) modified to supply 0, 11, 22, 44, 66, or 88 g(P) m^-3 (P0, P11, P22, P44, P66, P88). Phosphorus stress occurred in both cultivars at P0 and P11, with reduced plant growth and development. At P0, the lowest percentage of total biomass was directed toward reproductive growth. The root/shoot ratio was greatest at P0, reflecting greater dry matter partitioning to the root system. Growth of ‘San Luis’ was more sensitive to phosphorus stress than ‘Jupiter’. A greater percentage of total biomass was directed towards reproductive growth in ‘Jupiter’ than ‘San Luis’. Increasing P nutrition elevated leaf tissue P in both cultivars with highest leaf tissue P at P88. There were no differences in tissue P between P0 and P11 ‘San Luis’ plants, whereas P0 ‘Jupiter’ plants had the lowest tissue P. Low P-plants generally had the highest tissue N and lowest S, Mn, and B. In both cultivars, gas exchange was lowest at P0, as indicated by reduced stomatal conductance (g_s) and net photosynthetic rate (P_N). Internal CO₂ concentration and leaf-to-air vapor pressure difference (VPD) were generally highest with P-stressed plants. Phosphorus use efficiency, as indicated by P_N per unit of leaf tissue P concentration (P_N/P), was highest at P11. Generally, no P treatments exceeded the gas exchange levels obtained by P44 (full strength LANS) plants. Both P_N and g_s declined during reproductive growth in ‘San Luis’, which fruits more rapidly than ‘Jupiter’, whereas no reduction in gas exchange occurred with ‘Jupiter’. This revised version was published online in September 2006 with corrections to the Cover Date.     

Salinity tolerance of normal-fruited and cherry tomato cultivars

The salinity tolerances (NaCl) of 8 normal-fruited tomato cultivars (Lycopersicon esculentum Mill.) and 4 cherry tomato cultivars (L. esculentum var.cerasiforme) were determined by yield-substrate EC response curves, according to the Mass-Hoffman model, modified by van Genuchten and Hoffman (1984). The same model was used to determine the response curves of leaf dry-weight, stem dry-weight, and plant height against substrate EC and also between yield and leaf concentrations of Cl- and Na ions.According to the salinity-threshold (maximum EC-value without yield reduction) and slope (yield decrease per unit EC increase) parameters, determined from the yield-EC response curves, the cherry tomato cultivars were more salt-tolerant than the normal-fruited ones. However, on the basis of vegetative growth characters-EC response curves, cherry tomato cultivars and normal-fruited ones were similarly affected by NaCl.The ranking of the cultivars by their salinity tolerance, determined from the plots of yield vs. leaf concentrations of Cl- and Na ions, was the same as that evaluated from the yield vs. substrate EC plots.     

The response of two rice cultivars to external Na/Ca ratio

The response of the rice cultivars ‘M9’ and ‘M-201’ to nutrient cultures salinated at −0.4 MPa with varying ratios of Na and Ca was studied. Although the dry matter production of both cultivars was sensitive to the Na/Ca ratio, this correlation was significant only for M-201. Calcium nutrition was severely affected by the composition of the external solution, and the laminae exhibited Ca-deficiency symptoms at Na/Ca molar ratios of 78 and 198. Sodium concentration in the shoot decreased as the Na/Ca ratio in the external solution decreased. Patterns of Na and Cl distribution in the shoot tissues were similar; both ions were accumulated preferentially in the tillers and older leaves. The Na-induced inhibition of Ca uptake and transport appears to be more limiting to shoot growth of M9 and M-201 than Na toxicity per se.     

Response of tomato cultivars to salinity

The responses of five tomato cultivars (L. esculentum Mill) of different degrees of salt tolerance were examined over a range of 0 to 140 mM NaCl applied for 3 and 10 weeks. Judged by both Na and Cl accumulations and maintenance of K, Ca and Mg contents with increasing salinity, the most tolerant cultivars (Pera and GC-72) showed different responses. The greater salt tolerance of cv Pera was associated with a higher Cl and Na accumulation and a lower K content in the shoot than those found in the other cultivars, typical of a halophytic response to salinity. However, the greater salt tolerance of cv GC-72 was associated with a retention of Na and Cl in the root, restriction of their translocation to the shoot and maintenance of potassium selectivity under saline conditions. The salt tolerance mechanisms that operated in the remaining cultivars were similar to that of cv GC-72, as at first they excluded Na and Cl from the shoots, accumulating them in the roots; with longer treatment, the ability to regulate Na and Cl concentrations in the plant was lost only in the most salt sensitive cultivar (Volgogradskij), resulting in a massive influx of both ions into the shoot.The salt sensitivity of some tomato cultivars to salinity could be due to both the toxic effect of Na and Cl ions and nutritional imbalance induced by salinity, as plant growth was inversely correlated with Na and Cl contents and directly correlated with K and Ca contents. This study displays that there is not a single salt tolerance mechanism, since different physiological responses among tomato cultivars have been found.     

Inhibition of ethylene synthesis in tomato plants subjected to anaerobic root stress

Enhanced ethylene production and leaf epinasty are characteristic responses of tomato (Lycopersicon esculentum Mill.) to waterlogging. It has been proposed (Bradford, Yang 1980 Plant Physiol 65: 322-326) that this results from the synthesis of the immediate precursor of ethylene, 1-aminocyclopropane-1-carboxylic acid (ACC), in the waterlogged roots, its export in the transpiration stream to the shoot, and its rapid conversion to ethylene. Inhibitors of the ethylene biosynthetic pathway are available for further testing of this ACC transport hypothesis: aminooxyacetic acid (AOA) or aminoethoxyvinylglycine (AVG) block the synthesis of ACC, whereas CO²⁺ prevents its conversion to ethylene. AOA and AVG, supplied in the nutrient solution, were found to inhibit the synthesis and export of ACC from anaerobic roots, whereas Co²⁺ had no effect, as predicted from their respective sites of action. Transport of the inhibitors to the shoot was demonstrated by their ability to block wound ethylene synthesis in excised petioles. All three inhibitors reduced petiolar ethylene production and epinasty in anaerobically stressed tomato plants. With AOA and AVG, this was due to the prevention of ACC import from the roots as well as inhibition of ACC synthesis in the petioles. With Co²⁺, conversion of both root- and petiole-synthesized ACC to ethylene was blocked. Collectively, these data support the hypothesis that the export of ACC from low O₂ roots to the shoot is an important factor in the ethylene physiology of waterlogged tomato plants.     

Salinity-induced changes in essential oil,pigments and salts accumulation in sweet basil (Ocimum basilicum) in relation to alterations of morphological development

The objective of the project was to study salinity-induced effects on essential oil, pigments and salts accumulation in sweet basil (Ocimum basilicum, the cultivar Perrie) in relation to the alteration of plant morphological development and yield production. Hydroponically grown plants were exposed to one of six NaCl concentrations (1, 25, 50, 75, 100 and 130 mM NaCl). Inhibitory effects of salinity on biomass production of the shoot and the root, and area of individual leaves were apparent already under cultivation with 25 mM NaCl. Elevation of salinity from 1 to 100 mM NaCl induced 63% and 61% reductions in fresh and dry herb biomass production, respectively. The stress-induced reduction of foliage biomass sourced mainly from inhibition of leaf area development rather than reduction of internode and leaf number. Cl and Na concentrations in the leaves, stems and roots increased with elevation of NaCl concentration in the cultivation solution. While the extent of Cl accumulation was leaves>stems>roots, Na was largely excluded from the leaves and was preferentially accumulated in roots and the stems, potentially accounting for the moderate sensitivity of the leaf tissue to salinity. Salt stress increased the contents of essential oil and carotenoids in the leaves that may further account for the moderate sensitivity of sweet basil to salinity and suggest a potential for agro-industrial production. A twofold increase in both carotenoid concentration and the percent of essential oil in the fresh tissue was observed by elevation of the salinity from 1 to 130 mM NaCl. Overall, the stress induced increase of the percent of essential oil in the tissue in the salinity range 1–75 mM NaCl was about 50%, and thereby compensated for the similar reduction of biomass production in this salinity range, so that oil production on per plant basis was not reduced by salinity.     

Root apoplastic barriers block Na+ transport to shoots in rice (Oryza sativa L.)

Rice is an important crop that is very sensitive to salinity. However, some varieties differ greatly in this feature, making investigations of salinity tolerance mechanisms possible. The cultivar Pokkali is salinity tolerant and is known to have more extensive hydrophobic barriers in its roots than does IR20, a more sensitive cultivar. These barriers located in the root endodermis and exodermis prevent the direct entry of external fluid into the stele. However, it is known that in the case of rice, these barriers are bypassed by most of the Na(+) that enters the shoot. Exposing plants to a moderate stress of 100 mM NaCl resulted in deposition of additional hydrophobic aliphatic suberin in both cultivars. The present study demonstrated that Pokkali roots have a lower permeability to water (measured using a pressure chamber) than those of IR20. Conditioning plants with 100 mM NaCl effectively reduced Na(+) accumulation in the shoot and improved survival of the plants when they were subsequently subjected to a lethal stress of 200 mM NaCl. The Na(+) accumulated during the conditioning period was rapidly released when the plants were returned to the control medium. It has been suggested that the location of the bypass flow is around young lateral roots, the early development of which disrupts the continuity of the endodermal and exodermal Casparian bands. However, in the present study, the observed increase in lateral root densities during stress in both cultivars did not correlate with bypass flow. Overall the data suggest that in rice roots Na(+) bypass flow is reduced by the deposition of apoplastic barriers, leading to improved plant survival under salt stress.     

Grafting raises the salt tolerance of tomato through limiting the transport of sodium and chloride to the shoot

With the aim of determining whether grafting could improve salinity tolerance of tomato (Lycopersicon esculentum Mill.), and what characteristics of the rootstock were required to increase the salt tolerance of the shoot, a commercial tomato hybrid (cv. Jaguar) was grafted onto the roots of several tomato genotypes with different potentials to exclude saline ions. The rootstock effect was assessed by growing plants at different NaCl concentrations (0, 25, 50, and 75 mM NaCl) under greenhouse conditions, and by determining the fruit yield and the leaf physiological changes induced by the rootstock after 60 d and 90 d of salt treatment. The grafting process itself did not affect the fruit yield, as non-grafted plants of cv. Jaguar and those grafted onto their own root showed the same yield over time under non-saline conditions. However, grafting raised fruit yield in Jaguar on most rootstocks, although the positive effect induced by the rootstock was lower at 25 mM NaCl than at 50 and 75 mM NaCl. At these higher levels, the plants grafted onto Radja, Pera and the hybrid VolgogradskijxPera increased their yields by approximately 80%, with respect to the Jaguar plants. The tolerance induced by the rootstock in the shoot was related to ionic rather than osmotic stress caused by salinity, as the differential fruit yield responses among graft combinations were mainly related to the different abilities of rootstocks to regulate the transport of saline ions. This was corroborated by the high negative correlation found between fruit yield and the leaf Na(+) or Cl(-) concentrations in salt-treated plants after 90 d of salt treatment. In conclusion, grafting provides an alternative way to enhance salt tolerance, determined as fruit yield, in the tomato, and evidence is reported that the rootstock is able to reduce ionic stress.     

Efficient and simple plant regeneration via organogenesis from leaf segment cultures of persimmon (Diospyros kaki thunb.)

Summary An efficient and simple plant regeneration system via organogenesis from leaf segments of persimmon (Diospyros kaki Thunb.) cultivars ‘Fuyu’ and ‘Nishimurawase’ has been developed. The regeneration capacity was influenced by the culture vessels, gelling agents, plant growth regulators, and light conditions. Leaf explants taken from in vitro shoots were cultured on a modified Murashige and Skoog medium (MS1/2N), for 16 wk without transfer to fresh medium. Adventious shoots appeared after 4 and 8 wk in culture of ‘Nishimurawase’ and ‘Fuyu’ tissues, respectively. The culture of leaf explants in Erlenmeyer flasks with medium containing 4 g l⁻¹ agar enhanced shoot formation in comparison to media with increased agar concentrations. Optimal shoot regeneration was obtained with 5 mg l⁻¹ (22.8 μM) zeatin and 0.1 mg l⁻¹ (0.05 μM) indole-3-butyric acid (IBA) for ‘Nishimurawase’, and 10 mg l⁻¹ (45.6 μM) zeatin and 0.1 mg l⁻¹ (0.05 μM) IBA for ‘Fuyn’. Shoot regeneration frequencies in both cultivars were 100%, and shoot numbers per explant reached up to 9.2 for ‘Nishimurawase’ and 2.2 for ‘Fuyu’. Dark incubation during the first 4–5 wk was the most effective condition to successfully influence shoot regeneration in both cultivars. While dark incubation was essential for adventitious shoot formation by ‘Fuyu’, it was only slightly beneficial to ‘Nishimurawase’. More than 80% of the regenerated shoots rooted within 4 wk on hormone-free MS1/2N demium after having been dipped for 30 s in 250 mg l⁻¹ (1.1. mM) IBA solution.     

Photosynthesis parameters and activities of enzymes of oxidative stress in two young ‘Chemlali’ and ‘Chetoui’ olive trees under water deficit

The effects of water deficit on photochemical parameters and activities of superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase were investigated in two olive cultivars differing in drought tolerance — ‘Chemlali’ and ‘Chetoui’. After 30 days without irrigation, leaf water potential fell to −5.5 MPa that was accompanied by a marked decrease in net photosynthesis in ‘Chetoui’ olive cultivar. Maximal efficiency of PSII photochemistry (F_v/F_m) decreased slightly in ‘Chemlali’ (28 %) and substantially in ‘Chétoui’ (47 %). Both cultivars showed a similar decline (about 25 %) in the photochemical quenching coefficient, but only the drought-sensitive olive cultivar exhibited an enhancement (31 %) of non-photochemical fluorescence quenching under water deficit conditions. The quantum yield of electron transport decreased in both olive cultivars. ‘Chemlali’ showed a higher protection against oxidative stress, as judged from the lower levels of the malondialdehyde production. Catalase activity was higher in ‘Chetoui’. Glutathione reductase activity was increased similarly in both olive cultivars under water stress. Ascorbate peroxidase activity was enhanced in ‘Chemlali’ under water stress, but was unaffected in ‘Chetoui’. While, superoxide dismutase activity was inhibited in both cultivars under water stress, but higher activity was detected in ‘Chemlali’. Thus, the ability to increase ascorbate peroxidase and a higher superoxide dismutase activity might be an important attribute linked to the drought tolerance in ‘Chemlali’ olive cultivar.     

Differences in abscisic acid concentration in roots and leaves of two young Olive (<Emphasis Type="Italic">Olea europaea</Emphasis> L.) cultivars in response to water deficit

Differences in abscisic acid (ABA) accumulation between two olive cultivars were studied by enzyme-linked immunosorbent assay in roots and leaves, leaf water potential (Ψ_l), stomatal conductance (g _s) as well as photosynthetic rate (A) were also determined in well-watered (WW) and water-stressed (WS) plants of two olive cultivars ‘Chemlali’ and ‘Chetoui’. ‘Chemlali’ was able to maintain higher leaf CO₂ assimilation rate and leaf stomatal conductance throughout the drought cycle when compared with ‘Chetoui’. Furthermore, leaf water potential of ‘Chemlali’ decreased in lower extent than in Chetoui in response to water deficit. Interestingly, significant differences in water-stress-induced ABA accumulation were observed between the two olive cultivars and reflect the degree of stress experienced. Chemlali, a drought tolerant cultivar, accumulated lower levels of ABA in their leaves to regulate stomatal control in response to water stress compared to the drought sensitive olive cultivar ‘Chetoui’ which accumulated ABA in large amount.