Application of chlorophyll fluorescence for the diagnosis of salt stress in tomato “Solanum lycopersicum (variety Rio Grande)”

To study the response of tomato (Solanum lycopersicum cv. Rio Grande) to salinity, the effect on plant growth, water relations, stomatal conductance and Chlorophyll fluorescence was investigated. Tomato plants were grown in peat culture under controlled conditions and submitted during 28 days to saline stress ranging from 0 to 25, 50, 100, 150 and 200 mM of NaCl. At the end of the experiment period, plant growth was significantly decreased with increasing salinity.     

In vitro multiplication of statice plantlets using sugar-free media

Clumps of statice (Limonium latifolium) plantlets grown photomixotrophically were used as explants and cultured for 25 days on a sugar-free modified Murashige and Skoog (MS) medium in Magenta-type vessels with the number of air exchanges of the vessel (NAE) being 3.8 h⁻¹, at a photosynthetic photon flux (PPF) of 100 μmol m⁻² s⁻¹ and a CO₂ concentration of 1500 μmol mol⁻¹ in the culture room. A factorial experiment was conducted with three levels of 6-benzylaminopurine (BA) concentration, namely 0, 0.25 and 0.5 mg L⁻¹, and two types of supporting material, agar and Florialite (a porous material). The control treatment was a photomixotrophic culture using a sugar- and BA (0.25 mg L⁻¹) containing agar medium in the vessel with NAE of 0.2 h⁻¹, at a PPF of 50 μmol m⁻² s⁻¹ and a CO₂ concentration of 400 μmol mol⁻¹ in the culture room. Leaf area, chlorophyll concentration and net photosynthetic rate were greater in the sugar-free medium treatment with a BA concentration of 0.25 mg L⁻¹ and Florialite than those in the control treatment. The number of shoots and dry weight per clump in the sugar-free medium treatment were comparable to those in the control treatment. Among the sugar-free medium treatments, the number of shoots increased with increasing BA concentration, however, the leaf area, dry weight, chlorophyll concentration and net photosynthetic rate decreased with increasing BA concentration. The use of Florialite significantly enhanced the growth and root induction as well as net photosynthetic rate, compared with the treatments that use agar. These results indicated that sugar-free medium micropropagation could be commercially applied to the multiplication of statice plantlets.     

Response of apple trees to boron fertilization under conditions of low soil boron availability

The aim of the study was to examine the effect of boron (B) fertilization on the vegetative and the reproductive responses of apple (Malus domestica Borkh.) trees grown at low soil B availability. The experiment was carried out in 2005 under a greenhouse on 5-year-old ‘Jonagold’ apple trees/M.9 EMLA planted singly in 50-L containers filled with a sandy loam soil with hot water-soluble B concentration of 0.32 mg kg⁻¹. The trees were fertilized with B as foliar or soil application. Foliar B sprays were applied at the stage of pink bud, beginning of flowering, petal fall, and 10 days after flowering, at a solution concentration of 0.03%. Soil B fertilization was done at the bud break stage at a rate of 2 g per tree (27 mg B kg⁻¹ soil). The trees untreated with B served as the control. The results showed that soil B fertilization improved root development and tree vigor. Leaves of trees supplied with B to the soil had higher B concentration and chlorophyll, net photosynthetic rate, stomatal conductance, and activity of catalase and glutathione reductase than those of the control plants. Boron fertilization, regardless of application mode, increased fruit yield; the efficiency of foliar B sprays was higher than soil B application. Apple fruits of trees fertilized with B to the soil were bigger, more colored, richer in B, and had higher soluble solids concentration, and titratable acidity compared to those of the control trees.     

Response to drought and salt stress of lemon ‘Fino 49’ under field conditions: Water relations,osmotic adjustment and gas exchange

Drought and salinity are two of the most important factors limiting the lemon yield in south-eastern Spain. The effects of drought and salt stress, applied independently, on water relations, osmotic adjustment and gas exchange in the highest evapotranspiration period were studied to compare the tolerance and adaptive mechanisms of 13-year-old ‘Fino 49’ lemon trees, in immature and mature leaves. The study was carried out in an experimental orchard located in Torre Pacheco (Murcia). Three treatments were applied: Control, well-irrigated; drought-stress (DS), non-irrigated from 15th May to 7th July and salinity, irrigated with 30 mM NaCl from 1st March to 7th July. At the end of the experiment, only DS trees showed a decreased leaf stem water potential (Ψ_md). Under DS conditions, both types of leaf lost turgor and did not show any osmotic or elastic mechanism to maintain leaf turgor. Osmotic adjustment was the main tolerance mechanism for maintenance of turgor under salt stress, and was achieved by the uptake of Cl⁻ ions. Gas-exchange parameters were reduced by DS but not by salinity, stomatal closure being the main adaptive mechanism for avoidance of water loss and maintenance of leaf turgor. Salinity gave rise to greater Cl⁻ accumulation in mature than in immature leaves. The increase of proline in immature leaves due to DS indicates greater damage than in mature leaves.     

Moving lamps increase leaf photosynthetic capacity but not the growth of potted gerbera

To investigate the responses of leaf photosynthesis and plant growth to a moving lighting system, potted gerberas (Gerbera jamesonii H. Bolus ex J.D. Hook “Festival”) were grown under supplemental lighting in a greenhouse with either a stationary or a moving lighting system positioned above the benches. The stationary system consisted of a fixed high pressure sodium (HPS) lighting system, while the moving lighting system consisted of a moving HPS fixture attached to a cable system to move the light fixture back and forth over the crop. In both cases, the supplemental lighting was applied from 6:00 to 24:00 h with the same supplemental daily light integral (4.9 mol m⁻² day⁻¹). Moving lamps significantly increased leaf photosynthetic capacity as represented by light saturated net CO₂ exchange rate (NCER) (A_sat), light- and CO₂-saturated rate of NCER (A_max), maximum rate of Rubisco carboxylation (V_cmax), maximum rate of electron transport (J_max) and rate of triose phosphate utilization. However, in situ leaf NCER and stomatal conductance, leaf chlorophyll content index, leaf area, leaf thickness, fresh weight of plants were significantly lower under moving lighting than under stationary lighting. It is suggested that the reduced growth of plants under moving lighting might be due to (1) the overall lower light use efficiency of leaves under moving lighting than those under stationary lighting; (2) the slower response time of the photosynthetic system compared to the rate of change in light intensity under moving lighting.     

The response of different almond genotypes to moderate and severe water stress in order to screen for drought tolerance

In order to screen almond genotypes for drought tolerance, three different irrigation levels including moderate and severe stress (Ψs = −1.2 and −1.8 MPa respectively) and a control treatment (Ψs = −0.33 MPa) were applied for five weeks to six different cultivated almond seedlings. A factorial experiment was conducted with a RCBD which included 3 irrigations factors, 6 genotype factors and 3 replications. Seeds were prepared from controlled pollination of the bagged trees (after emasculation and flower isolation using isolator packets in the previous year). Genotypes included: homozygote sweet (Butte), heterozygote sweet (SH12, SH18, SH21 and White) and homozygote Bitter (Bitter Genotype). Leaf and root morphological and physiological traits including; midday relative water content, midday leaf (xylem) water potential, shoot dry weight and growth, total leaf area, leaf size, total leaf dry weight, specific leaf area, leaf greenness (SPAD), stomatal size and density, root and leaf nitrogen content and chlorophyll fluorescence were measured throughout the study. Results showed the six genotypes had different reactions to water stress but all genotypes showed an ability to tolerate the moderate and severe stresses and they showed different degrees of response time to drought stress. Almond seedling leaves could tolerate Ψ_w between −3 and −4 MPa in short periods. Water availability did not significantly affect stomatal density and size of young almond plants. The analysis of leaf anatomical traits and water relations showed the different strategies for almond genotypes under water stress conditions. Although almond seedlings even in severe stress kept their leaves, they showed a reduction in size to compensate for the stress effects. All genotypes managed to recover from moderate stress so Ψ_w = −1.2 could be tolerated well by almond seedlings but Ψ_w = −1.8 limited young plant growth. Leaf greenness, leaf size, shoot growth, shoot DW, TLDW and stomatal density were not good markers for drought resistance in almond seedlings. Root DW/LA, lower stomatal size and lower SLA might be related to drought resistance in cultivated almonds. Butte had the least resistance and White showed better performance during water stress while other genotypes were intermediate. Bitter seedlings showed no superiority in comparison with other genotypes under water stress conditions except for better germination and greater root DW which might make them suitable as rootstocks under irrigation conditions.     

Sennoside content and yield attributes of Cassia angustifolia Vahl. as affected by NaCl and CaCl2

Pot culture experiments were conducted to assess the extent of growth, photosynthetic capacity, sennoside concentration and yield attributes of Senna plant under the individual as well as combined influence of NaCl and CaCl₂. Six treatments, i.e. NaCl (80 and 160 mM), CaCl₂ (5 and 10 mM) alone and a combination of NaCl + CaCl₂ (80 + 10 and 160 + 10 mM) were given to the growing Senna plants at pre-flowering (45 DAS), flowering (75 DAS) and post-flowering (90 DAS) stages. Significant reductions were observed in pod biomass, leaf area, stomatal conductance, photosynthetic rate and sennoside concentration and yield, with each NaCl treatment. On the contrary, individual CaCl₂ treatments had a favourable effect. Under the effect of combination treatments, although these parameters were reduced, the extent of reduction was much less than one caused by NaCl treatments. The combined treatments thus mitigated the adverse effects caused by NaCl.     

Nitrogen best management practice for citrus trees: I. Fruit yield,quality, and leaf nutritional status

Elevated levels of nitrate-nitrogen (NO₃-N) in the surficial aquifer above the drinking water quality standard, i.e. maximum contaminant limit (MCL; 10 mg L⁻¹), have been reported in some part of central Florida citrus production regions. Soils in this region are very sandy (sand content >95%), hence are vulnerable to leaching of soluble nutrients and chemicals below the rooting depth of the trees. The objective of this research was to develop N and irrigation best management practices for citrus in sandy soils to maintain optimal crop yield and quality, and to minimize potential leaching of nitrate below the root zone. Six years of field experiment was conducted in a high productive (mean fruit yield > 80 Mg ha⁻¹yr⁻¹) >20-year-old ‘Hamlin’ orange trees [Citrus sinensis (L.) Osbeck] on ‘Cleopatra mandarin’ (Citrus reticulata Blanco) rootstock grown on a well drained Tavares fine sand (hyperthermic, uncoated, Typic Quartzipsamments) in Highland county, FL. Nitrogen rates ranged from 112 to 280 kg ha⁻¹ yr⁻¹ applied as fertigation (FRT), water soluble granular (WSG), 50:50 mix of FRT and WSG, and controlled-release fertilizer (CRF). Tensiometers were used to monitor the soil water content as a basis to schedule optimal irrigation. Fruit yield response over the entire range of N rates was greater for the FRT and WSG sources as compared to that for the WSG + FRT or CRF sources. Using the regression analysis of the fruit yield in relation to N rate, the optimum N rate appeared to be at 260 kg ha⁻¹ yr⁻¹. Based on fruit production response in this study, the N requirement for production of 1 Mg of fruit varied from 2.2 to 2.6 kg across four N sources. This study demonstrated an increased N uptake efficiency, as a result of best management of N and irrigation applications. The optimal N and K concentration in the 4–6-month-old spring flush leaves were 26–30, and 15–18 g kg⁻¹, respectively. However, fruit yield response showed no significant relationship with concentrations of P in the 4–6-month-old spring flush leaves over a range of 0.8–2.4 g kg⁻¹. The results of fate and transport of N in soil and in soil solution with application of different rates and sources of N, and components of citrus tree N budget, are reported in a companion paper.     

Nutritional quality of greenhouse lettuce at harvest and after storage in relation to N application and cultivation season

The effect of five levels of nitrogen fertilization on the growth and nutritional quality of Cos lettuce (Lactuca sativa L. cv. Parris Island) at harvest and after storage was studied during autumn and winter in South-West Greece. Plants were cultivated hydroponically in a greenhouse and the nitrate, chlorophyll and ascorbic acid (vitamin C) concentrations within the plant tissues were measured at harvest and following storage at 5 or 10 °C for 10 days. Nitrate accumulated in the leaves with increasing amounts of N within the nutrient solution and was higher in the winter than in the autumn. At the lowest N level (20 mg L⁻¹), the inner leaves accumulated more nitrate than the outer leaves, whereas at higher N levels (140, 200 or 260 mg L⁻¹) nitrate accumulation was higher in the outer leaves. Overall, the highest nitrate concentrations were detected in the petiole and the proximal end of the leaf, but at the lowest N application rate (20 mg L⁻¹) nitrate accumulated in the distal region of the leaf too. Although the nitrate concentrations within the leaves did not change significantly during 10 days storage at 5 or 10 °C, the chlorophyll and vitamin C concentrations decreased. Chlorophyll loss was higher in lettuce that was grown under low N levels and was higher at 10 °C than at 5 °C, but was reduced by enclosure of the lettuce in polyethylene film. It is concluded that the optimum N application rate for Cos lettuce grown hydroponically under cover during autumn and winter in South-West Greece, and in other areas with a similar climate, is 200 mg N L⁻¹ because at this N rate yield is satisfactory and leaf nitrate concentrations are below the maximum acceptable level for human consumption. Nutritional value (vitamin C concentration) and market quality (chlorophyll content) are highest at harvest and decrease during storage, but quality in terms of nitrate concentration does not change.     

Effects of leaf aging and light duration on photosynthetic characteristics in a cucumber canopy

Photosynthetic characteristics, chlorophyll index and leaf area were examined in selected leaves of cucumber (Cucumis sativus L. cv. Euphorbia). In the first experiment, plants of cucumber were grown horizontally at a lighting period of 20 h day⁻¹. Photosynthetic measurements in horizontally growing cucumbers showed that there was no decline in photosynthetic capacity when cucumber leaves are developing under good light conditions. In a second experiment, plants were grown in a traditional high-wire cultivation system under 20 h day⁻¹ lighting period until they reached final height and then exposed to different lighting periods (20 and 24 h day⁻¹) for 3 weeks. In stands of cucumber plants photosynthetic measurements showed that the lower leaves have a significant reduction in photosynthetic capacity due to reduced light conditions. Three weeks exposure to 24 h day⁻¹ lighting period reduced leaf area by 20%. Plant grown under continuous light had also lower chlorophyll index compared to plants grown under 20 h day⁻¹ lighting period.     

Root aeration improves yield and water use efficiency of tomato in heavy clay and saline soils

Water logging and salinity of the soil alter both the physical and biological environment of plant roots. In two experiments, we investigated the effects of imposed aeration on yield and the physiological response of tomato (Lycopersicon esculentum L.) variety Improved Apollo growing under protected conditions over a range of salinities (the salinity experiment), and under constant field capacity (FC) or drier soil conditions (the moisture experiment). Subsurface irrigation with aerated water (12% air in water) stimulated above-ground growth, and enhanced the reproductive performance through earliness for flowering and fruiting compared with the control. Fruit yield of tomato with aeration in the moisture experiment was increased by 21% compared with the control (4.2 kg versus 3.7 kg per plant), and the effect of aeration on fruit yield was greater in FC than in the drier treatment. Fruit yield was increased by 38% in saline soil due to aeration compared with the non-aerated control. Increasing salinity from 2 to 8.8 dS m⁻¹, and 10 dS m⁻¹ reduced fruit yield by 18% and 62%, respectively, but 4 dS m⁻¹ did not suppress yield. Aeration in both the experiments increased plant water use and water use efficiency (WUE), expressed as weight per unit of applied water. Biomass WUE was greater by 16% and 32% in the moisture and salinity experiments, respectively. The increased yield with aeration was also accompanied by an increased harvest index (HI) defined as the proportion of dry fruit biomass to total dry biomass, greater mean fruit weight, high fruit DM, and increase in leaf chlorophyll content and shoot: root ratio, and a reduced water stress index (computed from the difference between air and leaf temperature). The benefit gained from aerating irrigation water was not only observed under conditions where air-filled porosity may be low (e.g., in poorly structure sodic soils, or at field capacity in clay soils), but also in drier soils.     

Effect of organic and inorganic fertilizers applied during successive crop seasons on growth and nitrate accumulation in lettuce

A romaine-type lettuce (Lactuca sativa L.) cv. Corsica was cultivated during three successive crop seasons (late-spring, late-autumn and late-winter) in the same soil of an experimental greenhouse in S.W. Peloponnese, Greece. Seven long-term fertilization treatments were tested for their effect on plant growth and nitrate concentration in the external lettuce leaves. Treatments included: three different doses of organic fertilization (composted sheep manure) applied at the start of each crop season, three different doses of inorganic N fertilization applied via fertigation during each crop season, and a control treatment in which no fertilizer was applied. A drip irrigation system was used to water all plants. The highest nitrate levels were observed in the medium and maximum inorganic fertilization treatments (572–664 mg kg⁻¹) in all crop seasons. They were significantly higher compared to the respective organic fertilization treatments (253–435 mg kg⁻¹) and all other fertilization treatments (148–435 mg kg⁻¹). Crop season affected lettuce growth more than nitrate accumulation in the lettuce leaves: lettuce biomass production was the smallest and most uniform in the late-autumn season and did not respond to the fertilization treatments tested (ranging from 409 to 439 g plant⁻¹), while in the late-spring season biomass production was the highest and most variable (561–841 g plant⁻¹), it correlated with nitrate concentration in the leaves and in the medium and maximum inorganic fertilizer doses it significantly exceeded production from all other fertilization treatments (827–841 g plant⁻¹). Following the three crop seasons the residual availability of N, P and K was clearly enhanced in the soil receiving the organic compared to the inorganic fertilization. Nitrate concentration in lettuce leaves was far below the upper limits set by the European Commission in all fertilization treatments throughout the three crop seasons, a result attributed mainly to the sufficient level of light intensity and duration throughout the year in Southern Greece.     

Triacontanol stimulates nitrogen-fixation,enzyme activities,photosynthesis, crop productivity and quality of hyacinth bean (Lablab purpureus L.)

Hyacinth bean (Lablab purpureus L.) serves as a good source of vegetable proteins in human diet, and its seeds and pods contain as much as 20–28% protein. The bean contains tyrosinase enzyme, which has potential use for the treatment of hypertension. However, plant biological yield appears to be comparatively low in Aligarh soil in this region of India (Western Uttar Pradesh). A hypothesis was designed to address whether foliar application of triacontanol (TRIA) could enhance the crop productivity as well as crop quality. TRIA is known to be a potent plant growth promoting substance for many agricultural and horticultural crops. The hyacinth bean plants grown in soil containing pots, were sprayed with five concentrations of TRIA (10⁻⁰ (Control), 10⁻⁸, 10⁻⁷, 10⁻⁶ and 10⁻⁵ M) at 15-day intervals. The plant fresh and dry weights, leaf-area, number and dry weight of nodules per plant, total chlorophyll and carotenoid content, nitrate reductase activity, carbonic anhydrase activity, nodule-nitrogen content, leghemoglobin content and leaf N, P, K and Ca contents were analyzed at 60, 90 and 120 days after sowing (DAS). Net photosynthetic rate, transpiration rate and stomatal conductance were measured only at 90 DAS. The protein content plus carbohydrate and tyrosinase activity were analyzed in the seeds. Foliar spray of 10⁻⁶ M TRIA significantly stimulated most of the studied attributes. At the highest concentration (10⁻⁵ M TRIA), values of all attributes were significantly decreased at all three stages. For example, 10⁻⁶ M TRIA increased seed-yield and -protein content by 56.3 and 14.5%, respectively when compared to unsprayed plants. TRIA also stimulated the activity of tyrosinase when compared to the control plants.     

樱桃番茄气孔特征和气体交换参数对干旱胁迫的响应

以樱桃番茄为试材,采用基质培养的方法,使用不同浓度PEG 6000(0、5%、10%、15%)模拟干旱胁迫,研究了樱桃番茄叶片气孔特征、气体交换过程和叶绿素素含量对干旱胁迫的响应,以期为提升樱桃番茄在干旱地区土壤种植效率以及耐干旱品种的选育提供参考依据。结果表明:不同干旱处理对气孔长度、气孔周长、气孔面积、气孔宽度和气孔形状指数均产生显著的影响(P<0.05),除了气孔宽度外,均随着干旱胁迫的加剧而减少。随着干旱胁迫的加剧净光合反应速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)均呈现明显下降趋势(P<0.05)。但是,樱桃番茄叶片的细胞间CO_2浓度(Ci)随着干旱胁迫的加剧而升高,且与对照存在显著差异(P<0.05)。另外,干旱胁迫导致樱桃番茄叶片的水分利用效率(WUE)呈先减少后增加的趋势,但均小于对照。此外,樱桃番茄叶片的叶绿素含量随着干旱胁迫加剧呈现先上升后下降的趋势。表明干旱胁迫条件下樱桃番茄叶片主要通过调整单个气孔结构(气孔开度大小)及气孔分布特征(气孔密度和空间分布格局)及叶绿素含量来进一步优化气体交换效率。该研究结果不仅有助于深入理解干旱胁迫对樱桃番茄气孔特征、气体交换过程和叶绿素含量之间关系的影响机理,而且还将为提升樱桃番茄在干旱地区土壤种植效率以及耐干旱品种的选育提供参考依据。     

Priming abiotic factors for optimal hybrid Cymbidium (Orchidaceae) PLB and callus induction,plantlet formation,and their subsequent cytogenetic stability analysis

Abiotic factors affect the induction of PLBs and callus in hybrid Cymbidium Twilight Moon ‘Day Light’. The initiation and proliferation of new PLBs and callus could be achieved on NAA and kinetin, supplemented at 0.1 mg l⁻¹ each, respectively, both within 45–60 days. Bacto agar was found to be the most suitable solidifying agent for PLB induction, although a higher shoot fresh weight was obtained on Gelrite; a pH 5.3 was optimal while pH 4.5 caused 100% explant necrosis; coconut water, when supplied at 10–20% (v/v) resulted in a significant increase in the number of PLBs formed per PLB segment (23.1 versus 14.6 in controls) while a massive (almost four-fold) increase in fresh top weight occurred when PLB explants were placed in liquid culture, as a result of hyperhydricity; Fe-EDTA (1 mg l⁻¹) and activated charcoal (1 g l⁻¹) stimulated total fresh weight and PLB formation in the presence of PGRs; PLB formation decreased but total fresh shoot weight increased with the addition of niacin or myo-inositol, both vitamins. Dark-grown PLB-induced plants were etiolated and had longer internodes and higher fresh weight than light-grown control plants at 45 μmol m⁻² s⁻¹; at 15 μmol m⁻² s⁻¹ shoots were slightly etiolated, fragile, and PLB formation was scarce. RAPD and mtDNA analysis of all resultant PLBs, callus or plants showed them to be genetically identical, with comparable chlorophyll contents. Despite the detection of cytological variation between different plant parts, little variation resulted from abiotic factor treatment.     

Does root-sourced ABA play a role for regulation of stomata under drought in quinoa (Chenopodium quinoa Willd.)

The Andean seed crop quinoa (Chenopodium quinoa Willd.) is traditionally grown under drought and other adverse conditions that constrain crop production in the Andes, and it is regarded as having considerable tolerance to soil drying. The objective of this research was to study how chemical and hydraulic signalling from the root system controlled gas exchange in a drying soil in quinoa. It was observed that during soil drying, relative g_s and photosynthesis A_max (drought stressed/fully watered plants) equalled 1, until the fraction of transpirable soil water (FTSW) decreased to 0.82 ± 0.152 and 0.33 ± 0.061, respectively, at bud formation, indicating that photosynthesis was maintained after stomata closure. The relationship between relative g_s and relative A_max at bud formation was represented by a logarithmic function (r² = 0.79), which resulted in a photosynthetic water use efficiency WUE_Amax_/gs$\text{WU}{\text{E}}_{A_{\text{max}}/g_{\text{s}}}$ of 1 when FTSW > 0.8, and increased by 50% with soil drying to FTSW 0.7–0.4. Mild soil drying slightly increased ABA in the xylem. It is concluded that during soil drying, quinoa plants have a sensitive stomatal closure, by which the plants are able to maintain leaf water potential (ψ_l) and A_max, resulting in an increase of WUE. Root originated ABA plays a role in stomata performance during soil drying. ABA regulation seems to be one of the mechanisms utilised by quinoa when facing drought inducing decrease of turgor of stomata guard cells.     

Effect of genotype,explant size,position, and culture medium on shoot generation of Gerbera jamesonii by receptacle transverse thin cell layer culture

An unique procedure for the mass shoot propagation of Gerbera using receptacle transverse thin cell layer (tTCL) culture procedure was developed. Genotype, flower bud age, explant size, position of receptacle tTCLs and culture media were found to affect the success of culture. Ten interspecific crosses of Gerbera showed different shoot regeneration rates and callus induction via receptacle tTCL culture, all of which had shoot regeneration rates higher than 57%. Flower buds collected on the 10th day resulted in 91% shoot regeneration after 6 weeks of culture on basal MS medium [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol. Plant. 15, 475–497] supplemented with 0.02 mg l⁻¹ thidiazuron (TDZ), 0.8 mg l⁻¹ adenine and 10% (v/v) coconut water (CW). This was significantly higher than those from flower buds on the 7th and 14th days (22% and 54%), respectively. Shoot regeneration rate was the highest (94–100%) in the middle layers of the receptacle. For mass shoot propagation, shoot clusters were subcultured on half-strength MS medium supplemented with 0.5 mg l⁻¹ indole-3-butyric acid (IBA), 0.5 mg l⁻¹ 6-benzyladenine (BA) and 2.0 mg l⁻¹ kinetin after every 4 weeks. Plantlets formed when single shoots were cultured on half-strength MS medium containing 1 mg l⁻¹ IBA. All plantlets acclimatized well in the greenhouse.     

Molecular characterization and analysis of somaclonal variation in chrysanthemum cultivars using RAPD markers

Molecular characterization using RAPD analysis was carried out in eight cut flowers and two pot plant cultivars of chrysanthemum. Three of them (‘Refocus’, ‘Red Reagan’, and ‘Sheena Select’) were established in vitro and the occurrence of somaclonal variation was studied using the same molecular technique. Two induction media (MS + 0.1 mg l⁻¹ NAA + 0.1 mg l⁻¹ BA, and MS + 2.0 mg l⁻¹ IAA + 0.5 mg l⁻¹ Kinetin), and two proliferation media (MS + 0.1 mg l⁻¹ NAA + 0.2 mg l⁻¹ BA, and MS + 4.0 mg l⁻¹ IAA + 2.0 mg l⁻¹ Kinetin) were employed in order to evaluate the effect of the medium composition in the shoots’ stability. Likewise, the effect of the culture age was considered in assessing genetic stability. Monthly subcultures were carried out, identifying the origin and history of the shoots, throughout a nine-month proliferation period followed by acclimatization. Molecular markers were obtained in every subculture cycle and from the acclimatized plants. Only one shoot from the 7th subculture of the cultivar ‘Refocus’ showed a different band pattern. The use of RAPD for chrysanthemum cultivar characterization and somaclonal variation detection is discussed.     

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.     

Evaluation of the effectiveness of soil-applied plant derivatives of Meliaceae species on nitrogen availability to peach trees

We assessed the effect of soil-applied derivatives of melia (Melia azedarach L.) and neem (Azadirachta indica A. Juss) on nitrogen (N) soil availability, root uptake and peach (Prunus persica L.) growth. First we evaluated the effectiveness of experimentally prepared amendments made with fresh ground melia leaves or commercial neem cake incorporated into the soil as nitrification inhibitors, then we evaluated the effect of fresh ground melia fruits and neem cake on growth and N root uptake of potted peach trees, and on soil microbial respiration. Soil-applied fresh ground melia leaves at 10 and 20 g kg⁻¹ of soil as well as commercial neem cake (10 g kg⁻¹) were ineffective in decreasing the level of mineral N after soil application of urea-N as a source of mineral N, rather they increased soil concentration of nitric N and ammonium N. The incorporation into the soil of fresh ground melia fruits (at 20 and 40 g kg⁻¹) and neem cake (at 10 and 20 g kg⁻¹) increased N concentration in leaves of GF677 peach × almond (Prunus amygdalus) hybrid rootstock alone or grafted with one-year-old variety Rome Star peach trees. An increase in microbial respiration, leaf green color and plant biomass compared to the control trees were also observed. The Meliaceae derivatives did not affect, in the short term (7 days), N root uptake efficiency, as demonstrated by the use of stable isotope ¹⁵N, rather they promoted in the long term an increase of soil N availability, N leaf concentration and plant growth.