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In vitro preservation of transgenic tomato lines overexpressing the stress-responsive transcription factor SlAREB1 was studied by using slow growth and cryopreservation techniques. Slow growth preservation was performed by using different concentrations of sucrose (0, 100, 200, 300 mm) and abscisic acid (0, 4, 8, 12 μm) in Murashige and Skoog (MS) media, while cryopreservation was conducted by using encapsulation dehydration, V-cryoplates and seeds. Significant differences were observed between tested lines grown on MS media supplemented with 200 mm sucrose where transgenic lines overexpressing SlAREB1 showed improved growth when compared with negative control. The addition of abscisic acid (ABA) to the preservation media affected negatively transgenic lines growth and development when compared with ABA-free media. In encapsulation dehydration, non-cryopreserved transgenic lines overexpressing SlAREB1 pretreated in 0.8 M sucrose for 1 day and subjected to different dehydration periods showed significantly higher survival percentages when compared with negative control. For V-cryoplates technique, cryopreserved transgenic lines overexpressing SlAREB1 treated in 0.3 M sucrose for 3 days with or without cold acclimatization showed significantly higher survival percentages when compared with the negative control. Seed cryopreservation was performed successfully with a clear reduction in germination percentage in transgenic lines overexpressing high levels of SlAREB1. In conclusion, transgenic tomato lines overexpressing SlAREB1 were found to improve tolerance against different abiotic stresses associated with different in vitro preservation protocols.

Somatic embryogenesis was obtained from callus of wild arum (Arum palaestinum). Callus was induced from sterilized corm bud sprouts cultured on basal medium containing 4.4 μM 6-benzyladenine and 5.4 μM 1-naphthaleneacetic acid. Callus was maintained under dark conditions using basal medium with 4.4 or 8.8 μM 6-benzyladenine and 5.4 or 10.8 μM 1-naphthaleneacetic acid. The highest callus weight and most desirable callus phenotype were achieved using basal medium containing 8.8 μM 6-benzyladenine and 5.4 μM 1-naphthaleneacetic acid. Friable calli were cultured in the dark on basal medium containing 4.5 μM 2, 4-dichlorophenoxyacetic acid, 0.46 μM 6-furfurylaminopurine, 5.4 μM 1-naphthaleneacetic acid, and 1.7 mM proline to induce embryogenesis before transfer to regeneration medium. Embryos that developed on regeneration medium were transferred to medium minus plant growth regulators for germination. Ninety percent of the germinating embryos developed into rooted plantlets. Rooted plants were grown in the greenhouse and acclimatized successfully with a 95 % survival rate. This is the first report of successful somatic embryogenesis and plant regeneration in A. palaestinum.

Teucrium polium L. with the common name of Felty Germander is one of the plants flora that is widely used in folk medicine in many Middle East countries, it is an endangered plant species and must be highly considered for preservation. Cryopreservation of T. polium by vitrification and encapsulation-dehydration was successfully achieved in this study. Shoot-tips were excised aseptically from in vitro grown plants and incubated for 3 days on solid hormone free-Murashige and Skoog (HF-MS) media supplemented with 0.3 M sucrose under complete darkness at 24 ± 1 °C. In vitrification, shoot-tips were loaded in 0.4 M sucrose and 2 M glycerol for 20 min followed by desiccation with different combinations and concentrations of plant vittrification solution 2 (PVS2), before immersion in Liquid Nitrogen (LN). Whereas for the encapsulation-dehydration; shoot-tips were encapsulated in calcium alginate and dehydrated under laminar air flow cabinet for 0, 3, 6, or 9 h. A total of 60 % of the cryopreserved vitrified shoot-tips survived when desiccated in concentrated PVS2 solution for 20 min, whereas, 28 % of the cryopreserved vitrified shoot-tips were regrown after 20 min of desiccation by two step increase in PVS2 concentration. Complete survival were obtained for the non-cryopreserved encapsulated shoot-tips treated for 3 days in 0.5 M sucrose with MS media without or with 3 h of dehydration, whereas, only 20 % of the cryopreserved encapsulated shoot-tips were regrown. The procedures developed in this study are easy to handle and produced a high levels of shoot formation.

Artemisia herba-alba, called Shih is a medicinal herbal plant found in the wilds. The biodiversity of this plant is heavily subjected to loss because of heavy grazing, land cultivation and collection by people to be used in folk medicine. In the current study, two cryopreservation dependent techniques to conserve the shoot-tips of in vitro grown Shih were evaluated: encapsulation- dehydration and encapsulation- vitrification. Shoot-tips of Shih were encapsulated into sodium-alginate beads. In encapsulation- dehydration, the effect of sucrose concentration (0.5, 0.75 or 1.0 M) and dehydration period (0, 2, 4 or 6 h) under sterile air-flow on survival and regrowth of encapsulated shoot tips were studied. Maximum survival (100%) and regrowth (27%) rates were obtained when encapsulated unfrozen Artemisia herba-alba shoot tips were pretreated with 0.5 M sucrose for 3 days without further air dehydration. After cryopreservation the highest survival (40%) and regrowth (6%) rates were achieved when Artemisia herba-alba shoot tips were pretreated with 1.0 M sucrose for 3 days without further air dehydration. Viability of Artemisia herba-alba shoot tips decreased with increased dehydration period. In encapsulation-vitrification, the effect of dehydration of encapsulated Artemisia herba-alba shoot tips with 100% PVS2 for various dehydration durations (10, 20, 30, 60 or 90 min) prior to freezing was studied. After cryopreservation the dehydration of encapsulated and vitrified shoot tips with 100% PVS2 for 30 min resulted in 68% survival and 12% regrowth rates. Further conservation techniques must be evaluated to increase both survival and regrowth percentages.

A protocol for in vitro propagation of the wild germander (Teucrium polium L.) was developed. In vitro plants were developed from ex vitro axillary buds. Then, shoot tips were excised and established on Murashige and Skoog medium. Proliferation of shoots was tested with different levels of 6-furfurylaminopurin, 6-benzyladenine, or thiadiazuron. The highest proliferation of T. polium was obtained when 6-benzyladenine and 6-furfurylaminopurin were used at 2.0 and 1.6 mg l−1, respectively. Thiadiazuron gave the lowest response for shoot proliferation. Rooting was experimented at different levels of Indol-3-butric acid, Indol-3-acetic acid, or 1-naphthaleneacetic acid. 1-Naphthaleneacetic was the only growth regulator which promoted root induction. Rooted plants were acclimatized successfully with 75% survival and grown in the greenhouse. In vitro- and in vivo-grown plants were analyzed for essential oil production. In vitro-grown T. polium on MS medium supplemented with 6-benzyladenine and 1-naphthaleneacetic gave higher oil yield than that grown on hormone-free Murashige and Skoog medium. In vivo (wild)-grown T. polium produced different oil yield when collected in different months (April and October). β-caryophyllene, used as a marker compound in the essential oil, was identified and quantified by gas chromatography (GC) analysis. Gas chromatography/mass (GC-MS) spectrometry analysis was also used to identify other components of in vitro cultures and to compare with in vivo-grown plants.

A protocol for micropropagation of Arbutus andrachne from seeds was developed. Results indicated that none of the seeds cultured on Murashige and Skoog (MS) medium, with or without plant growth regulators (PGRs), germinated. Seeds soaked in 250 mg l−1 gibberellic acid (GA3) at 4°C for 3 days, then cultured on water-agar medium containing 2.0 mg l−1 GA3 exhibited 80–100% germination and developed into usable seedlings. Shoot proliferation was tested on MS or B5 medium containing different concentrations of cytokinin. No shoot proliferation was observed on PGR-free medium. Proliferation was more successful on MS than on B5 medium. On both media, the most successful proliferation was obtained using zeatin as a cytokinin type. Rooting was tested on MS medium containing different concentrations of auxin. Rooting failed on PGR-free medium and on medium containing indole-3-acetic acid (IAA), 0.25 or 0.5 mg l−1 indole-3-butyric acid (IBA), or 0.25, 0.5 or 2.0 mg l−1 α-naphthaleneacetic acid (NAA). Rooting (40%) was most successful with 1.0 mg l−1 NAA. Rooted plantlets exhibited 80% survival in all mixtures of peatmoss and perlite, and acclimatized plants were successfully grown in the greenhouse. Quantitative analysis of arbutin performed on in vivo and in vitro leaves using high-performance liquid chromatography (HPLC) revealed that in vivo leaves contained higher arbutin content (0.3–0.81% w/w) than in vitro leaves (0.09% w/w). The highest yield of arbutin in vivo was detected in leaves collected in August, and the lowest yield in leaves collected in December.

Physiological and biochemical responses of open-pollinated ‘Roma’ and dwarf F1 hybrid ‘Patio’ tomato (Lycopersicon esculentum Mill.) cultivars to in vitro induced salinity were examined in light of the possible contribution of ethylene to these symptoms. Salinity was induced by incorporating 0 (control), 50, 100, 150, or 200 mM NaCl into shoot culture media. Elevated salinity treatments significantly enhanced ethylene accumulation in the headspace and were accompanied by increased leaf epinasty in both cultivars. Growth, leaf cell sap osmolarity, leaf tissue viability and shoot soluble protein content were generally depressed with elevated salinity treatments, whereas electrolyte leakage, membrane injury, raffinose, and total sugars were concomitantly increased. Macronutrients N, P, K, Ca, Mg, and S decreased with elevated salinity in both cultivars and were accompanied by a significant increase in Na content and a sharp decrease in K/Na ratio. Tissue micronutrients, Fe, B, Zn, Mn, and Cu were generally decreased with elevated salinity especially at 100 mM or more. Incorporating ethylene inhibitors CoCl2 or NiCl2 at 5.0 or 10.0 mg/l into media supplemented with 100 mM NaCl significantly reduced ethylene accumulation in the headspace and prevented epinasty, but did not eliminate the negative impacts on growth and other physiological parameters caused by salinity treatment in either cultivar. Our results indicate that the increase in ethylene under salinity stress is not the primary factor contributing to salinity’s deleterious effect on tomato plant growth and physiology.

Cryopreservation of African violet via encapsulation-dehydration, vitrification, and encapsulation-vitrification of shoot tips was evaluated. Encapsulation-dehydration, pretreatment of shoot tips with 0.3 M sucrose for 2 d followed by air dehydration for 2 and 4 h resulted in complete survival and 75% regrowth, respectively. Dehydration of encapsulated shoot tips with silica gel for 1 h resulted in 80% survival but only 30% regrowth. Higher viability of shoot tips was obtained when using a step-wise dehydration of the material rather than direct exposure to 100% plant vitrification solution (PVS2). Complete survival and 90% regrowth were achieved with a four-step dehydration with PVS2 at 25°C for 20 min prior to freezing. The use of 2M glycerol plus 0.4M sucrose or 10% dimethyl sulfoxide (DMSO) plus 0.5M sucrose as a cryoprotectant resulted in 55% survival of shoots. The greatest survival (80–100%) and regrowth (80%) was obtained when shoot tips were cryoprotected with 10% DMSO plus 0.5M sucrose or 5% DMSO plus 0.75M sucrose followed by dehydration with 100% PVS2. Shoot tips cryoprotected with 2M glycerol plus 0.4M sucrose for 20 min exhibited complete survival (100%) and the highest regrowth (55%). In encapsulation-vitrification, dehydration of encapsulated and cryoprotected shoot tips with 100% PVS2 at 25°C for 5 min resulted in 85% survival and 80% regrowth.

Branched broomrape (Orobanche ramosa) is a holo-parasitic flowering plant that attaches to the root of its host, green plant, by means of a specialized structure known as haustorium. Following successful contact and penetration on susceptible plant root, complex tissue of Orobanche cells is formed which is known as the tubercle. Newly formed tubercles contain high activity ofindole-3-acetic acid (IAA). Triiodobenzoic acid (TIBA), as an inhibitor of IAA polar transport, was utilized to investigate the supply and requirement of auxin to the developing O. ramosa on tomato plant. There was no significant reduction in the incidence of O. ramosa per pot of different TIBA treatments. However, infection severity in terms of the number of O. ramosa shoots that emerged per plant and number of attachments per plant root system were significantly reduced by 60 % and 45 % on TIBA treated plants, respectively. Histo-logical studies revealed conspicuous delay in the initiation of xylem vessel differentiation inside tubercles of the TIBA treated tomato plants. Also, differentiated vessels showed thinner secondary wall deposition, and improper alignment within bundles inside those tubercles. They were wider and shorter in diameter in comparison to those of untreated plants. These findings were attri-buted to the short supply of IAA required for normal development, and to the xylem vessel differentiation of O. ramosa tubercles on infected tomato. Hence, this parasitic flowering plant seems to depend upon its host in its requirements for IAA, in a source to sink relation-ship.

A protocol for in vitro propagation of the wild three-lobed sage (Salvia fruticosa Mill.) (Synonym, Salvia triloba L.) was developed. Shoot tips were excised from in vitro seedlings and established on MS, Nitch and Nitch (NN), or B5 medium. For shoot proliferation, in vitro nodal and apical explants were cultured on MS medium containing 0.25–2 μM 6-benzylaminopurine (BA), 6-furfurylaminopurine (kinetin), or thidiazuron (TDZ). Proliferated microshoots were rooted on MS medium supplemented with 2.7–11.4 μM indole-3-butyric acid (IBA), indole-3-acetic acid (IAA), or α-naphthaleneacetic acid (NAA). Results indicated that shoots established at 100% regardless of media type, however, shoot height, nodes per shoot, and leaf number were highest for explants established on MS medium compared to NN or B5. Number and height of proliferated shoots, nodes per shoot, and leaf number were highest for nodal explants cultured on a medium containing 0.75 μM BA. Microshoots cultured on a medium supplemented with 2.7 μM IBA exhibited the highest rooting percentage compared to those cultured with IAA or NAA. Essential oil composition in microshoots and shoots of greenhouse-grown plants was determined by gas chromatography/mass spectrometry. The major essential oils detected in both plant materials were α-pinene, 1,8-cineole, camphor, and borneol. No α-thujone or β-thujone was detected. The content of essential oils, camphor, and borneol were higher in the microshoots than in shoots of greenhouse-grown plants.

Growth and nutrient acquisition in sour orange (Citrus aurantium L.) were studied under salt stress in vitro. Microshoots were transferred to Murashige and Skoog (MS) solid proliferation media containing 8.9 µM BA (6‐Benzyladenine) and 0.5 µM NAA (naphthaline acetic acid). Salinity was induced by incorporating different concentrations [0.0 (control), 50, 100, 150, 200, or 300 mM] of sodium chloride (NaCl) to the culture media. Microshoots were exposed to direct or gradual salinity shock. Slight reduction was obtained in growth (shoot length, shoot number, leaf number, and dry weight) when microshoots were directly exposed to NaCl stress from 0.0 to 150 mM. At 200 and 300 mM NaCl, growth parameters were adversly affected and microshoots died thereafter. Gradual NaCl shock was studied by transferring microshoots sequentialy every week to different NaCl concentraions (0.0, 50, 100, 150, 200, or 300 mM). Growth was monitored at each concentration until the end of the last week of incubation at 300 mM...

The objective of this study was to establish a cryopreservation protocol for sour orange (Citrus aurantium L.). Cryopreservation was carried out via encapsulation-dehydration, vitrification, and encapsulation-vitrification on shoot tips excised from in vitro cultures. Results indicated that a maximum of 83% survival and 47% regrowth of encapsulated-dehydrated and cryopreserved shoot tips was obtained with 0.5M sucrose in the preculture medium and further dehydration for 6 h to attain 18% moisture content. Dehydration of encapsulated shoot tips with silica gel for 2h resulted in 93% survival but only 37% regrowth of cryopreserved shoot tips. After preculturing with 0.5M sucrose, 80% of the vitrified cryopreserved shoots survived when 2M sucrose plus 10% dimethyl sulfoxide (DMSO) was used as a cryoprotectant for 20 min at 25°C. Survival and regrowth of vitrified cryopreserved shoot tips were 67% and 43%, respectively, when 0.4M sucrose plus 2M glycerol was used as a loading solution followed by application of 100% plant vitrification solution (PVS2) for 20 min. Increased duration of exposure to the loading solution up to 60 min increased survival (83%) and regrowth (47%) of cryopreserved shoot tips. With encapsulation-vitrification, dehydration with 100% PVS2 for 2 or 3 h at 0°C resulted in 50 or 57% survival and 30 or 40% regrowth, respectively, of cryopreserved shoot tips.

Responses of microshoot growth and micronutrient acquisition in bitter almond (Amygdalus communis L.) and sour orange (Citrus aurantium L.) to different iron (Fe) concentrations (7.45, 14.90, 22.35, or 29.80 mg L−1) added from FeEDTA or FeEDDHA were studied in vitro. Increasing FeEDTA concentration more than (22.35 mg L−1) or using FeEDDHA as Fe source resulted in a significant decrease in most growth parameters (microshoot length, proliferation, and dry weight). Compared to the control, Fe and manganese (Mn) acquisition by microshoots increased with increasing Fe concentration in all treatments. On the other hand, copper (Cu) and zinc (Zn) acquisition showed a decrease along with increasing Fe level. Generally, growth and micronutrient acquisition in both plants were higher when FeEDTA was used rather than FeEDDHA as Fe source except for Fe acquisition by bitter almond and Cu by sour orange. This could be attributed to the genetic variations between these species.

Silybum marianum L. is a wild medicinal herbal plant in Jordan. It is widely used in folk medicine due to its high content of silymarin. In vitro production of silymarin was experimented at different concentrations (0.0, 0.4, 1.0, 1.6 or 2.0 mg/l) of growth regulators (6-furfurylaminopurine (kinetin), 6-benzylaminopurine (BA), or 6-(gamma,gamma- Dimethylallylamino) purine (2ip)) and different concentrations (15, 30,45,60 mg/l) of carbon sources (glucose, fructose, and sucrose). HPLC (High Performance Liquid Chromatography) analysis was used to identify silymarin components. In vitro grown S. marianum on MS (Murashige and Skoo 1962) medium supplemented with (1.6 mg/l) kinetin and (0.1 mg/l) 1-Naphthaleneacetc acid (NAA) gave the highest silymarin content of (0.84%) silybin and (0.49%) silydanin as compared with cultures grown on hormone-free MS media which contained (0.36%) silybin and (0.30%) silydanin. In vitro grown S. marianum on MS medium supplemented with (2.0 mg/l) BA and 0.1 mg/l NAA yielded (0.67%) silybin and (0.37%) silydanin, while MS media supplemented with (1.0 mg/l) of 2iP gave (0.72%) silybin and (0.24%) silydanin. Otherwise, the in vivo (wild) grown shoots of S. marianum gave (1.07%) for silybin and (0.46%) sildyanin. Among carbon source, glucose at (45 g/l) gave (1.63%) of silymarin content. Results indicated a significant use of in vitro grown cultures for silymarin production.

Regeneration via somatic embryogenesis from callus was studied in ‘Nabali’ olive ( Olea europea L.). Among different explant sources (leaf blades, leaf petioles, hypocotyls of germinated seeds and roots of germinated seeds), roots gave the highest (46%) callus induction. Somatic embryogenesis was induced from root callus on embryogenesis induction medium (EIM) containing 5.0 μM 2,4-D, 0.5 μM kinetin and 5.0 μM NAA in darkness. Embryo regeneration was studied by transferring the callus from EIM to embryogenesis expression medium (EEM) containing different concentrations (0.0, 5.0, 10.0 and 15.0 μM) of 2-isopentenyladenine (2iP), BA, thiadiazuron (TDZ), zeatin or kinetin. Among the tested concentrations, 2iP at 10.0 μM outperformed the other growth regulators. 2,4-D at 5.0 μM in the EIM was satisfactory for embryogenesis induction. Sucrose at 0.2 M evoked higher embryogenesis than any other concentration of fructose and glucose in EIM, while sorbitol and mannitol at 0.1, 0.2 or 0.3 M reduced embryogenesis significantly and inhibited it totally at 0.4 M. Somatic embryos were rooted by transferring them to hormone-free medium (HFM). About 85% of embryos converted to rooted plantlets, 5% showed secondary embryogenesis and 10% were not developed and died. Rooted plantlets gave 95% survival when acclimatized ex vitro. Acclimatized plantlets developed into whole plants in the greenhouse and they were phenotypically similar.

Interactive effects of increased phosphorus (P) with salinity were studied at the microculture level of African violet (Saintpaulia ionantha). Increased P from 0.5 to 2.0 mM in the medium was very effective to mitigate the adverse effects of increased NaCl salinity (0.0, 50, 75, 100 mM). Growth (shoot height, and dry mass) was significantly reduced with increased salinity, whereas increasing P improved growth with elevated salt concentrations. Leaf osmolarity was decreased (more negative) with salinity effect and it was increased (less negative) by P treatments. Percent ash was increased with salinity and it was not highly affected by P. Root number and root length were significantly reduced with increased salinity and improved with increased P. The percentage of shoot content of nitrogen (N), P, calcium (Ca), potassium (K), and magnesium (Mg) were reduced with elevated salinity level and this reduction was less as P concentration increased in the medium. Sodium (Na) was significantly increased with impos...

Growth and micronutrient acquisition of some apple (Malus dometsica Borkh) varieties (Galla, MM 111, MM I06, M 26, and M6 York) were studied under gradually in vitro induced NaCl (from 0, 50 to 100 mM) salinity. Microshoot dry weight tended to increase with increasing salinity level in all varieties except in M 26 and M6 York as some decline was obtained at 100 mM NaCl. Shoot proliferation was decreased significantly in Galla at 100 mM NaCl and at both salinity levels in M 26 compared to the control (0 mM). A significant decrease in shoot zinc (Zn) acquisition with increasing salinity level was obtained in all varieties except MM 111. M6 York and MM 111 decreased their copper (Cu) acquisition significantly at 50 mM NaCl and then increased at 100 mM salinity level. Meanwhile, a significant decrease in Cu acquisition with increasing NaCl concentration was obtained in Galla and M 26. Galla, MM 106, and M 26 showed a significant increase in manganese (Mn) acquisition at 50 mM NaCl followed by a decre...

Somatic embryogenesis was achieved from callus, cell suspension and protoplast culture systems in the endemic black iris (Iris nigricans). Subculture of friable callus fragments on embryogenesis induction medium (EIM) containing 4.5 μM 2,4-dichlorophenoxyacetic acid (2,4-D), 0.5 μM kinetin, 4.5 μM 1-naphthaleneacetic acid (NAA) and 300 mg l-1 proline in the dark was necessary before transfer to regeneration medium (RM). Regeneration was studied by transferring friable callus fragments from EIM to RM containing (0.0, 4.5, 9.0, 13.5 μM) of either 6-benzyladenine (BA), 2-isopentenyladenine (2iP), zeatin or thidiazuron (TDZ) in combination with 0.49 μM indole-3-butyric acid (IBA), 0.45 μM 2,4-D. Maximum embryogenesis was obtained at 4.5 μM BA while zeatin and TDZ were not effective and embryogenesis did not occur with these treatments. Sucrose at 0.2 M was more effective for embryogenesis when compared to glucose or fructose. Growing cells in suspension culture on EIM containing 4.5 μM 2,4-D in combination with 0.2 M sucrose for four weeks and transferring cells to RM (containing 4.5 μM BA) gave significant embryogenesis with maximum number of embryos (3568 embryos/g cells). Using 4.5 μM 2,4-D in protoplast culture was necessary for the best protoplast division and colony formation. In all experiments, embryos developed on RM were transferred to hormone-free medium (HFM) and 90% converted to rooted plantlets. Produced plantlets gave 95% survival ex vitro. Plantlets developed to whole plants in the greenhouse and flowered.

In vitro and hydroponic intercropping systems were compared to examine the biocompatibility responses between ‘Fiona F1’ tomato (Lycopersicon esculentum Mill.) and ‘Spunta’ potato (Solanum tuberosum). The biological response of monocultures (with four plantlets per vessel) of each species (including change in medium electrical conductivity (EC), osmotic potential (OS), and tissue mineral element content) were directly compared to intercropped cultures of the following tomato:potato; 1:1, 2:1, 3:1, 1:2, 1:3, and 2:2 plantlets per vessel. The in vitro intercropping systems were performed using a Murashige and Skoog (MS) shoot proliferation medium (5.0 μM BA and 0.5 μM IBA). The parallel hydroponic intercropping system was performed using a Hoagland's solution. Monoculture of both species grows better in both systems than intercropped species. In the intercropped vessels, tissue mineral content decreased, medium pH became more acidic, OS decreased and EC increased as compared to monocultures in para...

Changes in medium pH, osmolarity (OS), and electrical conductivity (EC) were studied as a function of time in Murashige and Skoog (MS) liquid proliferation and rooting medium. Microshoots of wild pear (Pyrus syriaca), bitter almond, and ‘Spunta’ potato were targeted. Results indicated an acidic drift in pH in the different species on both proliferation and rooting medium. The EC was increased significantly with time whereas solution OS was decreased. Different species caused different effects on EC or OS depending on growth medium. The decline in OS in the proliferation medium was higher when pear and almond were grown, while on rooting medium the decline was sharper with potato and almond. The EC of proliferation medium was increased more when almond was grown; while on the rooting medium all species had similar effects. Under these saline conditions, low nutrient elements availability, poor plant growth, and nutrient element imbalance are expected. This study illustrates that the change in the ...

In vitro plantlets or microtubers (in vitro produced tubers) of ‘Spunta’ potato (Solanum tuberosum L.) were planted in a 3 soil: 2 peat moss: 1 sand substrate (by volume) amended with municipal solid waste (MS W) compost at 0, 10, 20, or 30 g 4‐1 L pot. Three months later, plant growth and tuber yield were evaluated and concentrations of shoot and tuber tin (Sn), arsenic (As), copper (Cu), zinc (Zn), nickel (Ni), lead (Pb), manganese (Mn), cadmium (Cd), and iron (Fe) were determined. Amending with MSW resulted in significant increases in concentrations of all tested metals in the substrate. Number of proliferated shoots of plants started from rooted plantlets was greatest at 10 g pot‐1 MSW, whereas shoot weight of plants started from microtubers was greatest at 10 and 20 g pot‐1 MSW. Tuber yield of plants started from rooted plantlets or microtubers was greatest at 10 or 30 g pot‐1 MSW, respectively. In all instances, amending with MSW at 30 g pot‐1 resulted in significant increases in concentrat...

Pigment producing in vitro cells of Vaccinium pahalae (ohelo) were tested for their ability to survive cryopreservation and retain pigment-production capacity after encapsulation-dehydration. Preculture of cells for 6 to 8 days in a medium containing 1.0 M sucrose was essential before dehydration. Reduction of bead water content before quenching in liquid nitrogen was highly correlated (r = 0.94) with increased survival rate in cells after cryopreservation. Dehydration of beads for 4 h was satisfactory for survival of cells. After quenching in liquid nitrogen, colored cells became pale, but pigment content was recovered once cells resumed growth. After three subcultures, cells regained their maximum capacity for pigment accumulation. The percentage of colored-to-total cell volume was not influenced by cryopreservation. Encapsulation-dehydration and cryopreservation did not diminish the capacity of cells to produce anthocyanins and other flavonoids.

Two Vaccinium genotypes growing in vitro (V. pahalae, ohelo, and V. myrtillus, bilberry) were examined under variable iron (Fe) and growth regulator regimes in order to elucidate the cause(s) of abnormal tissue formation. Two Fe sources (200, 300, and 400 μM Fe from FeEDTA and 150, 200 and 250 μM Fe from a combination of 100 μM FeEDTA supplemented with FeEDDHA) were used in combination with 12.3 μM 2iP (N‐[3‐methyl‐2‐butenyl]‐1‐H‐purine‐6‐amine), 18.2 μM zeatin, or 11.4 μM zeatin riboside. No hyperhydration and only trace chlorosis occurred in 2iP treatments, but growth was substandard, as reflected by significantly lower fresh and dry mass yields from shoots harvested after eight weeks. Although zeatin or zeatin riboside treatments induced greater shoot proliferation, the incidence of hyperhydration and chlorotic foliage was severe, especially for ohelo, in all treatments with supplemental FeEDTA. In FeEDTA‐supplemented media, exogenous zeatin riboside accumulated in foliage by the end of a grow...

Some factors which affect the in vitro propagation of wild pear ( Pyrus syrica ) were studied. Successful shoot proliferation was obtained on Murashige and Skoog medium supplemented with 1.5–2.0 mg 1 −1 benzyl adenine. Shoot height decreased to 1.8–1.6 cm and 1.3 cm with benzyl adenine (1.5–2.0 mg 1 −1 ) and zeatin (1.0–2.0 mg 1 −1 ), respectively. Benzyl adenine (at 2.0 mg 1 −1 ) and zeatin (at 1.5–2.0 mg 1 −1 ) enhanced callus formation. Indole butyric acid, indole acetic acid and naphthalene acetic acid induced in vitro rooting and a maximum of 72% rooting was achieved at 3.0 mg 1 −1 indole acetic acid. Neither activated charcoal nor polyvinylpyrrolidone influenced in vitro rooting. Ex vitro rooting was unsuccessful when treating shoot microcuttings with 0.0, 5.0, 10.0, or 15.0 mg 1 −1 indole butyric acid, indole acetic acid, or naphthalene acetic acid for l h and grown under intermittent water mist. An average of 95% survival was achieved when rooted microshoots were acclimatized under glass or plastic covers using intermittent mist in the greenhouse.

Ten certified potato cultivears (Ajax, Crystal, Kennebec, Labelle, Liseta, Mondial, Norchip, Signa Gold, Spunta and Vital) were grown at Jordan University Experimental Station in 1993. At harvest, tuber subsamples of each plot were analyzed for physical properties and chemical composition. Processed chips and fries were evaluated for appearance, color, aroma, texture, taste and overall acceptability. There was significant variation in tuber weight which ranged from 108.8 (Kennebec) to 148.7 g (Labelle), specific gravity which varied from 1.022 (Mondial) to 1.191 g/ml (Spunta), total solids which ranged from 18.9 (Ajax) to 23.9% (Vial), and chemical composition. Cultivars Liseta, Signa Gold, Mondial, Kennebec and Crystal produced highly acceptable chips and fries, while Ajax, Kennebec, Norchip and Spunta produced excellent fries only. All other cultivars produced less acceptable chips and fries.

Ohelo (V. pahalae Skottsb.) and bilberry (V. myrtillus L.) shoots were regenerated via direct organogenesis from whole leaves and leaf sections and also from hypocotyl explants of bilberry. Explants preincubated for 1 to 2 weeks in darkness yielded ≈75% regeneration frequencies and the highest number of regenerating shoots/explant on TDZ-supplemented media (0.9 to 2.7 μm). When 2iP or zeatin were substituted as the cytokinin source, frequencies of regeneration and shoot productivity were significantly lower. Explants held under constant illumination (no dark pretreatment) had significantly lower regeneration frequencies in all tested cytokinin-supplemented media. 2,4-D stimulated callus formation, but did not support regeneration from vegetative explants. Cells from callus and suspension cultures did not exhibit regeneration in any of the media that supported organogenesis from leaves. Regenerants were successfully micropropagated, although callus formation caused by zeatin and high 2iP levels interfered with shoot proliferation. Zeatin induced hyperhydricity in shoots from both species, but more severely in ohelo. Ex vitro rooting after treatment with 4.9 μm IBA or 5.4 μm NAA was 95% and 60% successful for bilberry and ohelo, respectively, and plants were readily acclimatized after an interval in a fog chamber. Bilberry microshoots also rooted in vitro in the absence of growth regulator treatment. Chemical names used: 1H-indole-3-butanoic acid (IBA); N-(3-methyl-2-butenyl)-1-H-purine-6-amine (2iP); 6-furfurylaminopurine (kinetin); 1-naphthaleneacetic acid (NAA); thidiazuron=1-phenyl-3-(1,2,3-thiadiazio-5-yl)urea (TDZ); 2,4-dichlorophenoxyacetic acid (2,4-D); 6-(4-hydroxy-3-methylbut-2-enylamino) purine (zeatin).

Successful in vitro multiplication of Capparis spinosa was achieved on Murashige and Skoog (MS) medium supplemented with benzyl amino purine (BAP) at 0.8 mg/L. The highest shoot length (35.6 mm) was obtained with the use of 0.4 mg/L BAP and 0.2 mg/L 1-naphthaleneanacetic acid (NAA). Kinetin at 2.0 mg/L produced a multiplication rate of 9.7 microshoots per explants with an average shoot length of 21.3 mm. In vitro rooting was successfully achieved on MS media supplemented with different concentration of indole-3-butyric acid (IBA), indole acetic acid (IAA) or NAA at various concentrations. However, rooting did not occur in the absence of IBA, IAA or NAA. A total of 85% survival was achieved when rooted explants acclimatized ex vitro using a mixture of 1 perlite: 1 peat. In another experiment, in vitro C. spinosa were successfully stored without serious losses by using MS medium supplemented with an appropriate concentration of osmoticum (sucrose, sorbitol, mannitol or glucose) at various concentration (0, 3, 6, 9 or 12%). Two types of plant material were used (in vitro plantlets and in vitro plantlets without tips). The results obtained show that the two type of plant material could be successfully maintained in vitro and optimum treatments were identified for each plant material. Further studies are still needed on medium term conservation to enhance the survival percentages of different plant material type. Key words: Clonal propagation, Capparis spinosa, medium term conservation, carbon source, roots formation.

Accumulation of essential oil in herbs depends on development stage. The aim of the present study was to investigate the effect of growth stages (pre-flowering and blooming stages) on the yield and composition of essential oil of thyme (Origanum syriacum L.). Three harvesting times (morning at 8 am, noon at 12 pm and evening at 6 pm) and two harvesting date (growth stage (pre-flowering stage) and blooming stage) were studied. Total yield of oil was higher in the plant at blooming stage in comparison with that at pre-flowering stage. According to the results of multivariate analysis, higher levels of volatile compounds were noticed during the blooming stage, except carvacrol, gamma-terpinene, alpha-terpinene and para-cymene that were found higher in pre-flowering stage. Unexpectedly, carvacrol as structural isomer of thymol depicts higher concentration at pre-flowering stage. Moreover, an accurate valid prediction equation of oil samples was built to predict the growth stage and quality control purposes of the oil. Key words: Carvacrol, essential oil, harvesting time, blooming stage, Origanum syriacum, thymol.

The growth and production of miniature dwarf tomato selection Lycopersicon esculentum ’Micro‐Tom’ plants grown from seedling to harvest in solution batph culture’ at four different NaCl salinity levels (2.4 [control, no NaCl], 7.6, 12.8, or 18 dS‐m‐1 solution conductivities) was monitored. Incremental reductions in canopy extent and shoot area of ‘Micro‐Tom’ were observed with increasing solution NaCl level. Root growth and shoot height were somewhat less responsive to imposed salinity. Fruit number, fruit size, and leaf tissue osmotic potential decreased as NaCl concentration increased. Fruit yield was highly correlated with total canopy and shoot area, but not with tissue osmotic or total water potential. ‘Micro‐Tom’ plants survived and continued fruit production at higher salinity levels despite reduced canopy growth. Treatment effects on vegetative growth and fruit production became more pronounced later in the growth cycle.

In vitro‐grown Chrysanthemum morifolium Ramat. cultivars Bright Golden Anne, Deep Luv, and Lucido were exposed to elevated mannitol, sucrose, or sorbitol concentrations to see if their response to this osmotic stress mimics that of in vivo plants enduring water deficit. The relative efficacy of the three osmotic agents at manifesting a response was also evaluated. Tissue osmolarity paralleled media mannitol and sorbitol concentration for all three cultivars. Shoot growth correspondingly decreased with increased osmolarity applied during the rooting phase. These responses generally resembled those of water‐stressed greenhouse plants. The degree of response varied with cultivar; ‘Lucido’ was the most sensitive and ‘Bright Golden Anne’ the least. Sucrose (metabolized by the plants) failed to elicit consistent osmotic stress symptoms; instead it enhanced both shoot and root growth. In vitro stress‐induced symptoms were produced in both proliferation and rooting stages, but consistent shoot growth red...

Successful clonal propagation of Stevia rebaudiana was achieved using microshoots as a primary step for in vitro conservation. Maximum proliferation was obtained on Murashige and Skoog (MS) medium supplemented with 1.5 mg L ‐1 benzyl amino purine and 0.2 mg L ‐1 indole-3-butyric-acid (IBA). Auxin increased rooting percentage of shoots at concentration of 0.4 mg L ‐1 IBA, indole-3-acetic-acid or naphthalene acetic acid and no rooting occurred without plant growth regulator. A survival of 90% was achieved when rooted explants were acclimatized in vivo in 1 soil: 1 perlite: 1 peat. In vitro S. rebaudiana shoots were successfully stored for up to 32 weeks on MS medium supplemented with an appropriate concentration of sucrose, sorbitol or mannitol, at 24 ± 2°C. After 32 weeks, 93.6% of the shoots were able to survive. Moreover, 89.3% of them were able to regrow when stored under light conditions. Cryopreservation by vitrif ication was successfully achieved (65.6% regrowth) when shoot tips were precultured on a medium supplemented with 0.4 M sorbitol for 2 d, followed by loading shoot tips with 80% concentrated plant vitrification solution 2 (PVS2) for 20 min; then dehydrated with 100% PVS2 for 60 min at 0°C prior to storage in liquid nitrogen. This procedure is easy to handle and produced a high levels of shoot formation. This protocol could be useful for longterm storage of S. rebaudiana germplasm.

Some factors that affect the in vitro conservation of wild pear (Pyrus syrica) microshoot cultures were studied. Sorbitol and mannitol at 0.2 to 4.0 M reduced growth significantly and extended the subculture intervals to 5 months when cultures where kept at 15°C. Increasing sucrose to 12% in the medium was not highly effective and the subculture intervals did not exceed 3.0 months. After 2 years of maintaining cultures on slow-growth medium, cultures grew slowly when transferred to fresh control medium. Shoots started to proliferate after three subcultures (6.0 weeks apart) on medium containing 1.0 mg/L BA and 0.1 mg/L NAA. New microshoots were rooted on medium containing 2.0 mg/L IBA and rooted microshoots gave 90% survival when acclimatized ex vitro under intermittent mist.