Effect of silicon on antioxidant and stomatal response of two grapevine (Vitis vinifera L.) rootstocks grown in boron toxic,saline and boron toxic-saline soil

We investigated individual and combined effects of B toxicity and salinity in the presence or absence of silicon on the shoot growth, concentrations of sodium (Na), chloride (Cl), boron (B) and silicon (Si), and stomatal resistance (SR), lipid peroxidation (MDA), proline accumulation, H₂O₂ accumulation and the activities of major antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT and ascorbate peroxidase, APX) activity grapevine rootstocks of 41B (V. Vinifera × V. Berlandieri) and 1103P (V. Berlandieri × V. Rupestris). Applied Si counteracted the deleterious effects of salinity and boron toxicity on shoot growth by lowering the accumulation of Na in 1103P, and B and Cl in the both rootstocks. Stomatal resistance, MDA, and the concentrations of H₂O₂ and proline were higher in the plants grown in conditions of B toxicity, salinity and their combination while applied Si lowered these parameters. Lowering SOD and CAT but increasing APX, Si treatment significantly affected the enzyme activities of both rootstocks. Based on the present work, it can be concluded that Si alleviates the adverse effects of salinity, B toxicity and combined salinity-B toxicity on grapevine rootstocks by preventing both oxidative membrane damage and translocation of Na and B from root to shoots and/or soil to plant, and also lowering the phytotoxic effects of Na and B within plant tissues. When considering the antioxidative response and membrane systems, it was concluded that the rootstock 1103P was responsive to Si under stress conditions. To our knowledge, this is the first report that Si improves the combined salt and B tolerance of grapevine grown under saline, B toxic, and B toxic and saline conditions which describes membrane related parameters and antioxidant responses.     

Silicon mediates changes to some physiological and enzymatic parameters symptomatic for oxidative stress in spinach (Spinacia oleracea L.) grown under B toxicity

The effect of exogenous silicon (Si) on the growth, boron (B) uptake, stomatal conductance, lipid peroxidation (MDA), membrane permeability, lipoxygenase activity (LOX), proline and H₂O₂ accumulation, non-enzymatic antioxidant activity (AA) and the activities of major antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT and ascorbate peroxidase, APX) of spinach were investigated under greenhouse conditions. Spinach plants were grown with 0 or 30 mg kg⁻¹ B combined with 0 and 150 mg kg⁻¹ Si. The severity of leaf symptoms of B toxicity was lower when the plants were grown with 150 mg kg⁻¹ Si. Silicon supplied to the soil with high B counteracted the deleterious effects of B on root and shoot growth. Application of B significantly increased B concentration in shoot and in root tissues. However, Si decreased B concentration in the shoots but increased it in the roots. Shoot tissues of spinach contained higher B than the roots in all treatments. Applied Si increased the Si concentration of the root and shoot. Stomatal conductance of the plants was decreased by B, but was increased by Si. The concentrations of H₂O₂ and proline were increased by B toxicity but were decreased by Si applied to plants. Boron toxicity increased the membrane permeability, MDA content and LOX activity of excised leaves of spinach. Applied Si ameliorated the membrane deterioration significantly. Compared with control plants, the activities of AA, SOD, CAT and APX in B-stressed plants without Si applied increased, and application of Si decreased their activities under toxic B conditions. Based on the present work, it can be concluded that Si alleviates B toxicity by preventing oxidative membrane damage and also translocation of B from root to shoots. To our knowledge, this is the first report on the effect of Si in improving B tolerance in spinach.     

The effects of low boron on growth,gas exchange,boron concentration and distribution of ‘Newhall’ navel orange (Citrus sinensis Osb.) plants grafted on two rootstocks

The effects of low boron (B) on plant growth, photosynthesis, B concentration and distribution of ‘Newhall’ orange (Citrus sinensis Osb.) plants grafted on either Trifoliate orange (Poncirus trifoliata (L.) Raf.) or Carrizo citrange [C. sinensis (L.) Osb. × Poncirus trifoliata (L.) Raf.] rootstocks were investigated. One-year-old plants of the two scion-rootstock combinations were grown for 183 days in sand:perlite (1:1, v/v) medium under greenhouse conditions. The plants were irrigated with half-strength Hoagland's nutrient solutions containing four B concentrations (0.01, 0.05, 0.10 and 0.25 mg l⁻¹). The growth of root, stem of scion and leaves was less affected by low B treatments when ‘Newhall’ scion was grafted on Carrizo citrange than on Trifoliate orange. Thus, the growth of scions under low B conditions was mainly depended on the rootstock used, i.e., Carrizo citrange-grafted plants were more tolerant to low B compared to the plants grafted on Trifoliate orange. Boron concentrations in all plant parts decreased significantly by decreasing the B supply in the nutrient solution. Leaves were the dominant sites of B accumulation and showed the greatest reduction in B concentration compared to the other plant parts, as B concentration in the nutrient solution decreased. Irrespective of the rootstock, B levels in the upper–younger leaves were substantially higher than in basal-older leaves when plants were exposed to low B concentrations (≤0.05 mg l⁻¹), suggesting that under such conditions B was preferentially translocated to upper–younger leaves to support their growth. Furthermore, B distribution in different plant parts indicated that there was a restriction in translocation of B from root to scion tissues (stems and leaves of scion) under conditions of limited B availability. In addition, low B treatments decreased leaf photosynthetic rate, stomatal conductance and transpiration rate but increased intercellular CO₂ concentration in the leaves of ‘Newhall’ plants, irrespective of the rootstock used.     

Changes in antioxidative system and membrane damage of lettuce in response to salinity and boron toxicity

Individual and combined effects of salinity and B toxicity on growth, the major antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; ascorbate peroxidase, APX) activities, ascorbic acid, proline, and H₂O₂ accumulation, and stomatal resistance (SR), malondialdehyde (MDA), membrane permeability (MP) and the concentrations of sodium (Na), chloride (Cl) and boron (B) of lettuce were investigated. Boron toxicity and salinity reduced growth of lettuce plants. Under B toxicity, B concentration of the plants was increased, but in the presence of NaCl, the concentration of B was significantly reduced. Sodium and Cl concentrations were increased in B + NaCl and NaCl treatments. Membrane damage was more pronounced in NaCl and B + NaCl treatments. Stomatal resistance of the plants was significantly increased by salinity treatments. The accumulation of proline and ascorbic acid was the highest in the B + NaCl treatment. In general, stress conditions significantly increased H₂O₂ and antioxidant enzyme (SOD, CAT and APX) activities. The present results indicate that stomatal closure is an important response of lettuce against NaCl and B + NaCl stress. Furthermore NaCl and B + NaCl toxicity-induced oxidative stress in lettuce resulting in lipid peroxidation and membrane damage. Increased antioxidant enzyme activities and also accumulation of ascorbic acid and proline are involved in order to overcome B- and NaCl-induced oxidative stress.     

Additional nitrogen fertilization affects salt tolerance of lemon trees on different rootstocks

Irrigation with saline water is one of the major problems in citrus crop in arid and semi-arid regions. Because rootstock and fertilization play an important role in citrus salt tolerance, we investigated the influence of the nitrogen fertilization and rootstock on salt tolerance of 2-year-old potted Fino 49 lemon trees. For that, trees grafted on Citrus macrophylla (M) or Sour orange (SO) rootstocks were watered for 12 weeks with complete nutrient solution containing either 0 mM NaCl (control, C), 50 mM NaCl (S), 50 mM NaCl with an additional 10 mM potassium nitrate (S + N), or 50 mM NaCl with a 1% KNO₃ (S + Nf) foliar spray application. Trees on M were more vigorous than trees on SO and saline treatments reduced leaf growth similarly in trees on both rootstocks. Trees on SO had a lower leaf Cl⁻ and Na⁺ concentration than those on M. Additional soil nitrogen (S + N) decreased leaf Cl⁻ concentration and increased leaf K⁺ concentration in salinized trees on both rootstocks. However, the salinity-induced reduction leaf growth was similar in S + N and S trees. This was due to osmotic effect, beside leaf Cl⁻ and Na⁺ toxicity, played an important role in the growth response of Fino 49 lemon to the salt stress. Additional foliar nitrogen in the S + Nf treatment also reduced leaf Cl⁻ concentration relative to the S treatment but trees from S + Nf treatment had the lowest leaf growth. Net assimilation of CO₂ (ACO₂$A_{\text{C}{\text{O}}_2}$), stomatal conductance (g_s) and plant transpiration were reduced similarly in all three salt treatments, regardless rootstock. Salinity reduced leaf water and osmotic potential such that leaf turgor was increased. Thus, the salinity-induced ACO₂$A_{\text{C}{\text{O}}_2}$ reductions were not due to loss of turgor but rather due to high salt ion accumulation in leaves.     

Zinc influence and salt stress on photosynthesis,water relations,and carbonic anhydrase activity in pistachio

A greenhouse study was conducted to evaluate the ameliorative effects of zinc (0, 5, 10 and 20 mg Zn kg⁻¹ soil) under saline (800, 1600, 2400 and 3200 mg NaCl kg⁻¹ soil) conditions on pistachio (Pistacia vera L. cv. Badami) seedlings’ photosynthetic parameters, carbonic anhydrase activity, protein and chlorophyll contents, and water relations. Zn deficiency resulted in a reduction of net photosynthetic rate and stomatal conductance. The quantum yield of photosystem II was reduced at zinc deficiency and salt stress. Zinc improved plant growth under salt-affected soil conditions. Increasing salinity in soil under Zn-deficient conditions, generally decreased carbonic anhydrase activity, protein, chlorophyll a and b contents. However, these adverse effects of salinity alleviated by increasing Zn levels up to 10 mg kg⁻¹ soil. Under increasing salinity, chlorophyll a/b ratio significantly increased. Zinc treatment influenced the relationship between relative water content and stomatal conductance, and between leaf water potential and stomatal conductance. It concluded that Zn may act as a scavenger of ROS for mitigating the injury on biomembranes under salt stress. Adequate Zn also prevents uptake and accumulation of Na in shoot, by increasing membrane integrity of root cells.     

Abscisic acid drenches can reduce water use and extend shelf life of Salvia splendens

Inadequate watering of potted ornamental plants during retail can cause a decline in quality as well as plant death. Abscisic acid (ABA) is the main phytohormone controlling plant responses to drought stress, including regulation of stomatal opening and closure. ABA may become available for the greenhouse industry in 2010. The objective of this study was to determine whether exogenous ABA applications can be used to induce stomatal closure and reduce water loss from the substrate. We drenched salvia (Salvia splendens F. Sellow. ex Roem & Shult.) ‘Bonfire Red’ with 50 mL of ABA solutions at concentrations of 0, 250, 500, 1000, and 2000 mg L⁻¹, after which plants were no longer irrigated. ABA drenches slowed down substrate water loss by reducing transpiration in a dose-dependent manner. Stomata closed within 3 h after ABA treatment, decreasing transpirational water loss. However, ABA drenches also caused abscission of lower leaves, resulting in approximately 60% leaf loss with 2000 mg L⁻¹ ABA. ABA drenches with 250 mg L⁻¹ and 500 mg L⁻¹ increased the shelf life of salvia ‘Bonfire Red’ by 3 days, without excessive leaf abscission. ABA can be used to increase the shelf life of salvia, but the lowest effective concentration should be used to minimize leaf abscission.     

Growth and flowering of black iris (Iris nigricans Dinsm.) following treatment with plant growth regulators

The effects of application method and concentration of gibberellic acid (GA₃), paclobutrazol and chlormequat on black iris performance were assessed. Plants (10 cm high, 4 ± 1 leaves) were sprayed with 125, 250, 375 or 500 mg L⁻¹ or drenched with 0.25, 0.5, 1 or 2 mg L⁻¹ GA₃. In a second experiment, the plants were sprayed with 100, 250, 500 or 1000 mg L⁻¹ or drenched with 0.25, 0.5, 1 or 2 mg L⁻¹ paclobutrazol. Other plants were sprayed with 250, 500, 1000 or 1500 mg L⁻¹ or drenched with 100, 250, 375 or 550 mg L⁻¹ chlormequat. In each experiment, the control treatment consisted of untreated plants. Results indicated that the tallest plants (37.3 cm) in the GA₃ experiment were those sprayed with 250 mg L⁻¹. The most rapid flowering (160 days after planting) occurred when a 375 mg L⁻¹ GA₃ spray was used, whereas flowering was delayed to 200 days using 1 mg L⁻¹ GA₃ drench. Drenching with 1 mg L⁻¹ GA₃ increased height of the flower stalk by 7 cm compared to the control. Though relatively slow to flower, plants drenched with 1 mg L⁻¹ GA₃ had long and rigid stalks, which were suitable as cut flowers. Number and characteristics of the sprouts were not affected by GA₃. All paclobutrazol sprays resulted in leaf falcation. A 500 or 1000 mg L⁻¹ paclobutrazol spray resulted in severe and undesirable control of plant height, drastic reduction in stalk height and weight, and delayed flowering. Plants drenched with 0.25 or 1 mg L⁻¹ paclobutrazol were suitable as pot plants. Chlormequat reduced plant height only at the highest drench concentration, which also reduced flowering to 70%. No leaf falcation was observed with GA₃ or chlormequat. Chemical names: ( ± )-(R*,R*)-beta-((4-chlorophenyl)methyl)-alpha-(1,1,-dimethylethyl)-1H-1,2,4,-triazol-1-ethanol (paclobutrazol); (2-chloroethyl) trimethylammonium chloride (chlormequat).     

An upper limit for elevated root zone dissolved oxygen concentration for tomato

It is well understood that insufficient oxygen within plant root zones can greatly diminish plant productivity. However, little is known about the effect of elevated root zone oxygen concentrations. Tomato (Lycopersicon lycopersicum Mill., cv. Trust) seedlings were grown in nutrient solutions containing dissolved oxygen (DO) concentration ranging from 5.3 to 40 mg L⁻¹ for 4 weeks. There were no visible symptoms observed on the leaves or stems in any of the treatments. Leaf chlorophyll content was higher in the 40 mg L⁻¹ treatment than with 20 and 30 mg L⁻¹ DO treatments. Two weeks from the start of the experiment, roots in the 40 mg L⁻¹ treatment exhibited stunted growth, became thicker, and had fewer side and fine roots compared to roots in the lower levels of DO treatment. Almost all the measured growth parameters (fresh and dry weights of root, stem, and leaf, leaf area, stem diameter) were significantly reduced in plants grown in the 40 mg L⁻¹ treatment compared to plants in the lower level of DO treatments, except that the plant height increased with the increasing DO concentration. Root respiration increased linearly with increasing DO concentration; however, there was no effect on leaf net CO₂ exchange rate. It is suggested that it was safe to enrich root zone DO to as high as 30 mg L⁻¹, although the growth benefit was minor by increasing DO from ambient air saturated level (∼8.5 mg L⁻¹) to 30 mg L⁻¹. Higher than 30 mg L⁻¹ could cause reduction in tomato plant growth.     

Rootstock influences the response of pistachio (Pistacia vera L. cv. Kerman) to water stress and rehydration

Pistachio cultivation requires the use of rootstock because grafting is the only form of vegetative propagation. The main commercial rootstocks are Pistacia integerrima L., Pistacia atlantica Desf., Pistacia terebinthus L. and Pistacia vera L. Pistachio is considered to be a drought and saline-resistant crop; however, there is little information describing varietal responses of rootstocks to water stress. Some studies have suggested that P. terebinthus L. is the most drought and cold resistant rootstock. The effect of the rootstock on the water relations of the grafted plant is crucial for improving crop performance under water stress conditions and for developing the best irrigation strategy. This work studied the physiological response to water stress of pistachio plants (P. vera L. cv. Kerman) grafted onto three different rootstocks P. terebinthus L., P. atlantica Desf. and a hybrid from crossbreeding P. atlantica Desf. × P. vera L. Plant physiological responses were evaluated during a cycle of drought and subsequent recovery in potted plants. Parameters measured were soil moisture, trunk diameter, leaf area, leaf number, leaf and stem dry weight, stem water potential, leaf stomatal conductance. The results showed different responses of cv. Kerman depending on the rootstock onto which it had been grafted. The hybrid rootstock was associated with a higher degree of stomatal control and reduced leaf senescence compared to P. atlantica and P. terebinthus, despite being associated with the most vigorous shoot growth. P. terebinthus enabled very effective stomatal control but was also associated with the most rapid leaf senescence. P. atlantica was associated with less vigorous shoot growth and similar levels of water stress as occurred with the others rootstocks under conditions of high evaporative demand, which was associated with lower stomatal control. The selection of the most effective rootstock choice for different environmental conditions is discussed.     

Direct regeneration of plants derived from in vitro cultured shoot tips and leaves of three Lysimachia species

The regenerability of three ornamental species—Lysimachia christinae, Lysimachia rubinervis and Lysimachia nummularia ‘Aurea’, were investigated using in vitro leaves and shoot tips. 6-Benzylaminopurine (BAP) and α-naphthalene acetic acid (NAA) added to Murashige and Skoog (MS) medium were tested for their effect on organogenesis. On the medium, shoot regeneration occurred directly without callus formation. In these species, L. christinae developed the highest regeneration rate and numbers of shoots/explant from shoot tips (100%, 12.25) and leaf bases (100%, 13.01) on the MS medium containing 3.0 mg l⁻¹ BAP and 0.1 mg l⁻¹ NAA. For L. rubinervis, the highest shoot induction rate and number of shoots/explant were obtained from shoot tip (100%, 16.87–17.20) on the MS medium with 0.1 mg l⁻¹ NAA and 3.0–5.0 mg l⁻¹ BAP. L. nummularia ‘Aurea’, however, showed the highest regeneration rate and number of shoots/explant (100%, 12.73) from leaf bases on MS medium supplemented with 1.0 mg l⁻¹ BAP and 0.1 mg l⁻¹ NAA. All in vitro shoots rooted well on half macronutrient MS medium containing 0.1 mg l⁻¹ NAA. After acclimatization, transplanted plantlets grew normally and flowered in the field.     

Regeneration and characterization of plants derived from leaf in vitro culture of two sweet orange (Citrus sinensis (L.) Osbeck) cultivars

In the current work attempts were made to investigate culture of leaf explants derived from in vitro seedlings of two sweet orange (Citrus sinensis (L.) Osbeck) cultivars, Bingtangcheng and Valencia. Effects of several factors, including culture medium, lighting condition, explant age and genotype on regeneration response were examined based on three parameters, percentage of explants producing shoots, mean number of shoots per explant and shoot forming capacity. Culture of the explants on shoot-inducing media (SIM) composed of MT salts supplemented with different growth regulators gave rise to disparate shoot regeneration, in which SIM₁ (MT + 0.5 mg L⁻¹ BA + 0.5 mg L⁻¹ Kinetin + 0.1 mg L⁻¹ NAA + 3% sucrose + 0.8% agar, pH 5.8) was shown to be the most effective medium for direct induction of shoots from leaf explants. Highly significant difference in the response of shoot bud regeneration was noted between the two cultivars, with Bingtangcheng being more responsive than Valencia. Culture of explants from fully developed leaves led to better shoot regeneration capacity in comparison to undeveloped ones. However, the two lighting conditions used herein did not cause significant difference in shoot regeneration. Phenotypic observation and randomly amplified polymorphic DNA (RAPD) analysis confirmed that all the regenerated plants from both genotypes were genetically identical to their donor plants, suggesting absence of detectable genetic variation in the regenerated plants. The data presented here demonstrated that direct initiation of plants from leaf explants has been successfully accomplished. To our knowledge, this is the first report on direct regeneration of shoots from leaf explants in Citrus, which will provide an alternative source for citrus genetic manipulation in the future.     

Impact of exogenous salicylic acid on the growth,antioxidant activity and physiology of carrot plants subjected to combined salinity and boron toxicity

Previous studies have shown that salicylic acid (SA) plays a role in the response of plants to salt and osmotic stresses. Therefore, an experiment was conducted to investigate the impact of exogenous salicylic acid on the growth, physiology and antioxidant activity of carrot (Daucus carota L. cv. Nantes) grown under combined stress of salinity and boron toxicity. The treatments consisted of salt (control, NaCl, and Na₂SO₄), boron (−B: 0 and +B: 25 mg B kg⁻¹) and salicylic acid (−SA: 0 and +SA: 0.5 mmol SA kg⁻¹). The diameter of the storage root was increased by NaCl salinity in the absence of B toxicity, however, it was increased by Na₂SO₄ salinity under B toxicity. For the storage root yield, NaCl salinity was more toxic than Na₂SO₄ salinity. With its role in plant growth regulation, SA application positively affected the storage root dry weight, S concentration, carotenoids and anthocyanin content and increased the total antioxidant activity (AA) of the shoot and storage root. SA application regulated proline and toxic ion (B, Cl) accumulation in the storage root and shoot. This study reports the long term effects of SA under stress conditions and reveals that SA was not as effective as in alleviating abiotic stress as reported in the literature conducted with short-term studies. That means long-term effects of SA would be significantly different from its short-term effects.     

Clonal propagation of Rosa clinophylla Thory. through axillary bud culture

A protocol for clonal propagation of Rosa clinophylla, a rare and endangered species but very important for breeding purposes had been standardized through in vitro axillary bud culture. Although cytokinins alone were able to induce shoot buds, but their proper growth and number could be increased only when they were used in combination with GA₃. However, there was shoot tip necrosis and leaf fall in the proliferated shoots. AgNO₃ at 58.85 μM proved effective to avoid shoot necrosis and yellowing of leaves. Activated charcoal (AC) at 250 mg l⁻¹ was found necessary at all the stages of shoot multiplication as well as rooting. Ninety percent rooting could be achieved in 1/2 MS medium supplemented with 4.92 μM IBA and 250 mg l⁻¹ AC. Rooted plantlets were hardened and transferred to the field successfully with 80% survival rate.     

High frequency early flowering from in vitro seedlings of Dendrobium nobile

Dendrobium nobile Lindl. is a popular temperate Chinese orchid commonly marketed as a traditional medicinal plant. Seedlings of Dendrobium nobile Lindl. produced floral buds (33.3–34.8%) precociously on a defined basal medium (1/2 MS) containing paclobutrazol (PP₃₃₃) at 0.5 mg L⁻¹ or thidiazuron (TDZ) at 0.1 mg L⁻¹ within 4 months of culturing. The frequency of floral buds formation can be further increased to 95.6% by growing seedlings in a PN (PP₃₃₃ 0.3 mg L⁻¹ + NAA 0.5 mg L⁻¹)-containing medium followed by transfer onto 1/2 MS medium with PP₃₃₃ and TDZ (PP₃₃₃ + TDZ). However, flower developed was deformed under 25 °C but it developed fully when grown in a lower temperature regime (23 °C/18 °C, light/dark) for 45 days. Under optimal condition, in vitro flowering was observed about 6 months after seed sowing.     

Embryo germination and proliferation in vitro of Juglans regia L.

This paper presents the optimal culture conditions for the in vitro embryo germination and proliferation of Juglans regia L. rootstock cv. Peralta, selected by the IMIDA fruticulture team. J. regia L. rootstock cv. Peralta is characterised by its resistance to salinity, lime and by its vigour. The first experiment determined the best culture medium for in vitro embryo germination. The Murashige and Skoog medium (MS; Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissues cultures. Physiol. Plant. 15, 473–497), the medium developed by our team for walnut (NGE), the Lloyd and McCown medium (WPM; Lloyd, G., McCown, B., 1981. Commercially feasible micropropagation of mountain laurel, Kalmia latifolia, by the use of shoot tip culture. Proc. Plant Prop. Soc. 30, 421–427) and that of Driver and Kuniyuki (DKW; Driver, J.A., Kuniyuki, A.M., 1984. In vitro propagation of Paradox walnut rootstock. HortScience 19 (4), 507–509), were compared, all without growth regulators. The best germination percentage was obtained in WPM (81%) with significant differences between the different media. In the second experiment, the optimal benzylaminopurine and indole butyric acid concentrations were determined for the proliferation stage of the explants obtained in the first experiment. The proliferation rates obtained varied from 0 in the medium without cytokinins to 6 in the medium with 0.5 mg l⁻¹ BAP. The cluster proliferation quality and other parameters studied indicate that the optimal treatment was 0.5 mg l⁻¹ BAP.     

In vitro proliferation from axillary buds and ex vitro protocol for effective propagation of Syringa × hyacinthiflora ‘Luo Lan Zi’

As a precondition for lilac mass propagation, the optimal shoot-multiplication medium for Syringa × hyacinthiflora ‘Luo Lan Zi’ was ascertained mainly based on clustered microshoot inducement and large leaf area establishment in 6-benzyladenine (BAP) (1.00 mg L⁻¹) and zeatin (Z) (0.10 mg L⁻¹) combination. Medium supplied with lower level of BAP (0.50 mg L⁻¹) and auxin (IAA) (0.25 mg L⁻¹) was not suitable for lilac shoot proliferation, but it could be competent for long-term preservation of the un-rooted shoots so that subsequent proliferation culture could be carried out at anytime. In addition, excess height growth which resulted in low transplanting survival rate was effectively controlled by decrease in node number when paclobutrazol (PBZ) was applied in rooting medium at a concentration of 1.00 mg L⁻¹ after taking into account the effects on shoot height, rooting, persistent leaf area and PBZ carry-over. An important overwintering treatment was to use a plastic chamber covering for plants in the greenhouse prior to field planting to ensure adequate biomass of stem and underground parts not only in the current growing season but also in the subsequent years.     

Cercis siliquastrum L., Malus trilobata Schneid and Acer syriacum Boin and Gaill water use as affected by two fertilizer rates

There has been an increased demand for landscaping plants in Lebanon as a result of numerous reconstruction projects. Sustainable landscape regulations have created a need for regionally adapted taxa, especially those with low water requirements. Therefore, water use of container-grown plants and the impact of fertilization on water use were studied in the following native species: Cercis siliquastrum L. (six mother trees), Malus trilobata Schneid (two mother trees) and Acer syriacum Boin and Gaill (one tree). Two-year-old containerized seedlings were grown at The Ohio State University (Columbus, USA) under two fertilizer rates: 25 or 100 mg N L⁻¹ of 21 N–3.1 P–5.9 K water soluble fertilizer. Water use estimates were made by saturating the containers early in the morning, allowing them to drain for 1 h, weighing them and re-weighing approximately 5 h later. Although there were differences in seedling heights, those grown at 25 mg N L⁻¹ were taller than those at 100, there were few differences in water use per seedling. In August, Cercis seedlings grown under 100 mg N L⁻¹ had higher height adjusted water use (g water cm⁻¹ height h⁻¹, a method for standardizing water use among different sized plants) than those grown under 25 mg L⁻¹. However, there were no differences in height adjusted water use in September attributed to fertilizer rates. In September, Acer seedlings had higher water use cm⁻² leaf surface area under 25 than 100 mg N L⁻¹. There were no differences in water use among the progeny from the six Cercis mother trees. However, the seedlings from one Malus tree had higher water use cm⁻² leaf surface area than those from the other tree, even though the extant trees were separated by less than 20 m.     

Factors affecting mango (Mangifera indica L.) protoplast isolation and culture

The major factors influencing protoplast isolation and culture of mango (Mangifera indica L.) cv. Kensington Pride were investigated. The resultant protocol was used to compare plating efficiency among 4 mango cultivars. Most responses differed between proembryonic masses (PEMs) and leaf sources. Protoplast yields of 15.22 × 10⁶ g⁻¹ from PEMs and 8.68 × 10⁶ g⁻¹ from greenhouse-derived leaves were obtained in a solution of 0.7 M mannitol CPW plus 1.5% cellulase, 1% hemicellulase and 0.75% macerozyme for PEMs or 0.5 M mannitol CPW plus 1.5% cellulose, 1% hemicellulase and 1.5% macerozyme for leaves. Culture in Ca-alginate beads with initial plating densities (IPD) of 2.5 × 10⁴ Pp mL⁻¹ for PEMs and 2.5 × 10⁵ for leaves gave the highest plating efficiencies (FPE). For PEMs 1 mg L⁻¹ 2,4-d and 3.5 mg L⁻¹ kinetin gave an FPE of 2.85% whereas lower kinetin (2 mg L⁻¹) plus 0.5 mg L⁻¹ 6-BAP was most effective for leaves (FPE of 2.12%). Most protoplast mortality occurred during the first week of culture and was more severe in liquid culture. In Ca-alginate beads, protoplast survival at 14 days was higher for PEMs (30%) than leaf (21%) as was the frequency of cell division (17.6% compared to 13.6%). PEMs protoplasts continued development through embryogenesis to in vitro plantlet regeneration whereas leaf protoplasts underwent cell division up to 40-cell colonies but failed to proceed further. For PEMs, polyembryonic cvs. Kensington Pride and Keow Savoey produced higher FPE (1.95%) than monoembryonic cvs. Tommy Atkins and Keitt (1.75%). There was no effect of cultivar for leaf protoplasts.     

‘Swingle’ citrumelo propagation by cuttings for citrus nursery tree production or inarching

‘Swingle’ citrumelo [Citrus paradisi MacFaden × Poncirus trifoliata (L.) Raf.] has been extensively used as a rootstock in several citrus growing regions of the World, including Southern Brazil where ‘Rangpur’ lime (Citrus limonia Osbeck) is still the predominant variety despite being affected by several important pathogens. In this case, ‘Swingle’ citrumelo is used to produce nursery trees to establish new orchards or to be inarched in adult and healthy groves in order to change the rootstock. We report herein a system to produce trees on ‘Swingle’ citrumelo more rapidly by budding onto non-rooted cuttings, as well as assessing potential to rapidly multiply ‘Swingle’ through rooting of non-budded cuttings. Therefore, two potential products are described: budded trees and rooted rootstock cuttings. ‘Valencia’ sweet orange [Citrus sinensis (L.) Osbeck] was budded at different heights on cuttings derived from eight-month old rootstocks. Grafted and additional non-budded cuttings were then treated with indole-3-butyric acid (500 mg L⁻¹) or left untreated before rooting. Three types of cuttings were evaluated: softwood, semi-hardwood and hardwood. The use of nursery trees derived from pre-budded hardwood cuttings of ‘Swingle’ citrumelo is an alternative grafting method on this cultivar. Softwood cuttings with one leaf pair were considered the most adequate material for rapid multiplication of ‘Swingle’ citrumelo by cutting. This could be particularly useful for inarching production or conventional budding after transplant of cutting-derived rootstocks.