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Ochradenus baccatus Delile is a wild medicinal plant that produces several natural compounds with medicinal benefits. In this study, microshoots of Ochradenus baccatus were exposed to osmotic stress conditions consisting of Murashige and Skoog solid media (MS) media that containing different osmotic agents (sugar types) at a range of concentrations (including 0.1, 0.2, 0.3 or 0.4 M). The purpose was to test their effects on microshoots’ growth and antimicrobial activities against selected strains of bacteria and one strain of fungi. It has been found that growth parameters (including shoot length and proliferation) of Ochradenus baccatus microshoots declined with increasing sugar concentration in the media, but at the highest concentrations of 0.4M mannitol, the micorshoots could not survive. Generally, aqueous extracts of the stressed microshoots were more effective against the tested microbial strains than the methanolic extracts in most experiments. Staphylococcus aureus was found to be the most affected microbe to both extract types. Also, exposing the microshoots to osmotic stress had improved antimicrobial powers in both extracts types. Aqueous extract of microshoots that pre-grew in media with (0.4 M sucrose) was interestingly found to inhibit growth of Staphylococcus aureus and Candida albicans with minimal inhibitory concentration (MIC) values of (0.195, 0.78 mg/ml). These values were similar to those obtained from the antibiotic treatments. Other biotechnological techniques like genetic transformation are suggested to be also used for production of elite strains of Ochradenus baccatus with super antimicrobial potential.

Nanotechnology is one of the most important modern sciences that has integrated all sectors of science. Nanotechnology has been applied in the agricultural sector in the last ten years in pursuit of increasing agricultural production and ensuring food security. Plant biotechnology is an essential science that is concerned with plant production. The use of nanotechnology in plant biotechnology under controlled conditions has facilitated the understanding of important internal mechanisms of the plant biological system. The application of nanoparticles (NPs) in plant biotechnology has demonstrated an interesting impact on in vitro plant growth and development. This includes the positive effect of the NPs on micropropagation, callus induction, somatic embryogenesis, cell suspension culture, and plant disinfection. In addition, other biotechnology processes, including the genetic transformation of plants, plant conservation, and secondary metabolite production have improved by the use of NPs. Furthermore, nanotechnology is used to improve plant tolerance to different stress conditions that limit plant production. In this review article, we attempt to consolidate the achievements of nanotechnology and plant biotechnology and discuss advances in the applications of nanotechnology in plant biotechnology. It has been concluded that more research is needed to understand the mechanism of nanoparticle delivery and translocation in plants in order to avoid any future hazardous effects of nanomaterials. This will be key to the achievement of magnificent progress in plant nanobiotechnology.

Plant biodiversity expresses one of the principal natural resources for all nations as it provides food, medicine, and shelter for all living besides its important role in balancing ecosystems and mitigating climate change. Jordan has paid great attention to its unique heritage of plant biodiversity since the foundation of the country, especially in terms of conservation. Notable conservation efforts have been made by Jordanian governments during the first hundred years of the country's foundation. These efforts were highly fruitful thanks to the unlimited governmental support to all projects related to the conservation of Jordan Flora. So, on the occasion of the first centenary of the country’s foundation, this review article was prepared to highlight some of the efforts made by Jordan during the past hundred years to conserve this national wealth of plant biodiversity.

Paronychia argentea is a wild medicinal herbaceous herb that contains many secondary metabolites. It grows wild in Jordan under harsh high salinity soil conditions. This study was conducted to investigate the effect of salinity stress on growth, biochemical responses, and ion uptake of in vitro Paronychia argentea micro plants. Our results revealed a clear reduction in all growth parameters in response to an increase in salinity level used (0, 25, 50, 100, 200 mM). When the growth medium was supplemented with 200 mM NaCl; the fresh and dry weights were reduced by 40% compared with the control treatment. Moreover, micro plant uptake of phosphorus calcium and potassium ions decreased with increasing salinity levels. The Mg++ content was increased by 10% at the 25 mM level of NaCl compared with control, while it declined by 20% at 200 mM of NaCl. On the other hand, Na+ uptake increased with increasing salinity levels and reached a maximum value of (18328 ppm/D. W) at 200 mM NaCl. Moreover, proline content increased in response to salinity level to record maximum value (80.8 µmole/g) at 200 mM NaCl compared to (30.4 µmol/g) obtained in the control. Quercetins content in P. argentea micro plants was increased by 1.7-fold in concentration in response to salinity at 50 mM of NaCl, while an about 7-fold increase in the concentration of isorhamnetin content was obtained due to salinity at 100 mM.

Solanum villosum is a wild distinguished medicinal herb that has few studies describing its medicinal properties with no sustainable production and conservation studies. In this study, in vitro cultures of microshoots and callus of Solanum villosum were multiplied in vitro on Murashige and Skoog media supplemented with different growth regulators. The MS media that have 2,4 Dichlorophenoxy acetic acid (2,4-D) and Kinetin at the levels of (2.0, 1.2 mg/L; respectively) were used for the induction and multiplication of callus that yielded the highest fresh weight of (6.03 ± 0.15) g. Cryopreservation was applied successfully to seeds using the dehydration technique and for both microshoots and callus using v-cryoplate technique. The highest regrowth rate was obtained in callus (95 ± 0.13) after pre-culturing callus on 0.3 M sucrose for 5 days. The most inhibited bacteria strains by microshoots extracts were, Klebsiella pneumoniae and Staphylococcus epidermidis (30 ± 0.25 and 25 ± 0.17 mm; respectively) using the disk diffusion assay. In MIC method; The extract obtained from microshoots achieved best results of MIC values (9.77 and 39.06 (µg/ml) against (Micrococcus luteus and Klebsiella pneumonia; respectively). A sustainable protocol of callus culture and cryopreservation protocol for in vitro Solanum villosum has been set for the first time which makes this research novel. Furthermore, promising results of antimicrobial activity for both in vitro and wild Solanum villosum were obtained.

An efficient protocol is outlined for rapid and mass micropropagation of Ficus carica L. (fig). Shoot tips (5 mm) were obtained from mother plants stock grown on half strength Murashige and Skoog (½ MS) medium with the addition of 30 g/L sucrose. For shoot multiplication Benzyl amino purine (BAP) and kinetin produced differences number of new shoot per plant and shoot height. BAP at 0.4 mg/L in combination with 0.2 mg/L indole-3-butyric-acid (IBA) produce maximum in vitro propagation rate, with 4.2 shoots per ex-plant. Root initiation was experimented on MS medium containing different concentrations of mg/L, IBA, IAA (Indole-3-acetic-acid) (IAA) or Naphthalene acetic acid (NAA). Highest number of root (4.3) was resulted when 1.5 mg/L IBA was used. After acclimatization in a mixture of (1 soil: 1 perlite: 1 peat) survival rate of 80% was achieved. For in vitro conservation of F. carica was experimented as microshoots were stored for 40 weeks on MS medium containing different sucrose concentration. Medium supplemented with 3% sucrose gave the highest regrowth (89%) at 24 ± 2 °C. Culture grew slowly when transferred to new fresh medium after the storage periods.

In recent years, higher education around Arab universities has witnessed, in many cases, a dramatic diminution in its quality of delivered product and industry-related research. This, inherently, would normally lead to degraded performance in the socio-economic ecosystem together with an associated deterioration in the economies of the countries involved. Additionally, this has had an adverse impact upon further industrial growth, which, in turn, caused stagnation and/or downturns of the economies of the constituent countries. Furthermore, the competitiveness of Arab economies has hit a brick wall due to the speed at which Western and South-East Asian industrial economies have grown. This has worsened the prevailing economic situation in many Arab countries even further and to catch up with the industrial sphere is a matter that has become something of the impossible! Smaller economies around the world have traditionally grown and prospered (emerged) as they leveraged certain niches that would put them on the map of integration with other world economies and survived mainly by making significant exports where the trade balance would stand in their favour. As the higher-education sector in the Arab world never exhibited any witnessed integration with the local industries of these countries, and where real (heavyweight) industries never emerged, a situation has arisen where real industrial competitiveness of the Arab world never surfaced. And when one would argue for the possibility of opting towards one form or another of an ICT industry, people are faced with the fact that countries that arrived to the scene much earlier, including Ireland, India, and China, compounded with the complication of an ocean of software freelancers, left the Arab world with little room for a real global industrial role. To that end, one sole approach remains to be addressed by the Arab world; one that would loosen up the grip of international economic export in the face of primary Arab industrial exports. To elicit real economic growth for Arab economies in the face of all the aforementioned, usually requires out-of-the-box approaches to help create some level of a global demand for Arab work products. Here, the way to go (and may be the only) would be to adopt innovation as a culture for a productive academic/industrial integration in these countries. This chapter will address the innovation ecosystem around the Arab world, in both industrial establishments and academic institutions and will propose one possible framework to help snatch the academic sector around the Arab world from its state of stagnation. It will also address ways of helping the industrial sector move forward and investigate effective ways to create a productive ecosystem for academic/industrial integration. This would leverage the inherent power of innovation towards creating start-ups, spinoffs, SMEs, and, therefrom, industrial establishments that the Arab world will someday pride itself with. Along the way, the chapter will address the prevailing governance structures of academic regulations to help the academic ecosystem bolster efforts in support of innovation and entrepreneurship throughout the Arab world.

Solanum nigrum L. is a medicinal plant of solanaceae family with distinguished therapeutic properties. Traditionally, S. nigrum . had been used as an anti-tumorgenic, antioxidant, hepatoprotective, diuretic, and antipyretic agent. The most important alkaloid member in this plant is solanine. Therefore, this study was conducted to utilize tissue culture techniques for the enhancement of solanine production in the in vitro grown cultures of this promising neglected plant. For callus growth and development experimental part, the highest callus growth parameters (callus diameter (21.4 mm) and callus fresh weight (2202.4 mg) were obtained in callus grown on Murashige & Skoog MS media supplemented with 2,4-Dichlorophenoxyacetic acid (2.0 mg·L -1 ) plus 1.5 mg·L -1 Thidiazuron . Similar trend was also obtained in cell suspension culture experiment, as maximum growth was recorded at similar hormone combination. Moreover, High-performance liquid chromatography analysis revealed that, solanine was affected by growth regulator type and concentration. The highest solanine levels were obtained when the explants were treated with 6-benzylaminopurine at level of 2.0 mg·L -1 , as solanine content reached up to (2.61, 1.53 mg.g -1 ) for callus and cell suspension, respectively, while, microshoot contained the highest solanine (4.52 mg.g -1 DW) at 6-benzylaminopurine level of 1.6 mg.L -1 . Additionally, carbon source had positively affected solanine level, where 0.2 M sucrose resulted in production of the highest amounts (3.13, 2.03 and 1.20 mg.g -1 DW) of solanine in microshoots, callus and cell suspension, respectively. Also, exposing microshoots and callus to light intensity of (100 µmol.m -2 s -1 ) yielded the highest solanine content (4.03 and 1.26 mg.g -1 DW, respectively),while the lowest solanine levels (1.50 and 0.48 mg.g -1 DW) were observed in plant material exposed to the lowest light intensity treatment (25 µmol.m -2 s -1 ) .

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.

□ Growth and nutrient acquisition of tomato (Lycopersicon esculentum L.) cv ‘Amani’ were studied under induced salt stress in Hoagland's solution. The plants were treated for 37 days with salinity induced by incorporating different concentrations [0.0 (control), 50, 100, 150, or 200 mM] of sodium chloride (NaCl) to the nutrient solution. Slight reduction was obtained in growth represented by (shoot length and number, leaf number, and dry weight) when seedlings were directly exposed to NaCl stress from 0.0 to 100 mM. At higher concentrations (150 or 200 mM), growth parameters were adversely affected and seedlings died thereafter. Elevated salinity significantly reduced crude protein and fiber in shoots and roots. Tomato shoot and root contents of potassium (K), iron (Fe), and ash were reduced significantly in response to increased levels of salinity. Tissue contents of sodium (Na) and chloride (Cl) increased with elevated salinity treatments.

Callus was induced from seeds of Colchicum hierosolymitanum Feib inoculated on the surface of MS media supplemented with 0.45 µM 2, 4-dichlorophenoxyacetic acid under dark conditions. Then callus was cultured on MS media supplemented with 4.52 µM 2, 4-dichlorophenoxyacetic for growth and maintenance. Friable callus from the fourth generation was transferred to liquid MS media supplemented with 0.54 µM 1-naphthaleneacetic acid to form cell suspension. Cells were successfully subcultured every 27 days on the same liquid media supplemented with 0.54 µM 1- naphthaleneacetic acid. Higher concentration (9 µM) of 6-benzyladenine with 0.45 µM 2, 4-dichlorophenoxyacetic acid resulted in higher cells fresh weight, while 1-naphthaleneacetic acid combinations with 6-benzyladenine had no effect on cell growth. On the other hand, the time for subculturing the cells into new fresh liquid media was determined to be after 27 days of incubation. (-) -Colchicine was identified in callus and cell suspension of C. hierosolymitanum by performing HPLC analysis against standard. Different ratios of NH +4 : NO -3 were used to study their effect on (-)-colchicine content, the highest colchicine content of 0.070 mg g -1 DW was obtained at 30 mM NH 4 + of total nitrogen. Colchicine alkaloid was highest, 0.090 mg g -1 DW, at 0.1 M of sucrose after 4 weeks incubation. (-) - Colchicine alkaloid was not detected in callus grown on sucrose free media. Maximum production of colchicine, 0.235 mg g -1 DW, was obtained in callus extracts of 60 days old callus grown under dark conditions. Cell suspension had 0.012 mg g-1 DW (-) -colchicine from suspended cells grown under dark. (-) -Colchicine content of callus incubated under dark (0.095 mg g -1 DW) was higher than light (0.070 mg g -1 DW) condition.

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.

The present study aims at developing a reliable protocol for in vitro propagation and cryostorage shoot tips of Stevia rebaudiana using a vitrification method. The highest multiplication rate (8.9) was achieved on Murashige and Skoog (MS) medium supplemented with 1.5 mg/L benzyl amino purine (BAP) and 0.2 mg/L Indole-3-butryic acid (IBA). For cyropreservation of shoot tips in liquid nitrogen, several pretreatment trials prior to cryopreservation were conducted to enhance shoot tips recovery after storage in liquid nitrogen. Culturing plantlets in vitro for four weeks in MS medium containing 30 g/L sucrose, excision of shoot tips; preculture of shoot tips on media supplemented with 0.4 M sorbitol for 2 d, followed by loading shoot tips with 80% PVS2 for 20 mint, then dehydrated with a highly concentrated vitrification solution (100% PVS2) for 60 min at 0°C prior plunge into liquid nitrogen produced the highest re-growth (up to 68.8%) post thawing at 45°C for 2 min. After six months post cryopreservation none of the growth parameters had been significantly affected by the cryopreservation process. The procedure developed in this study is easy to handle and produced high levels of shoot formation. Thus, the protocol is applicable for long-term storage of S. rebaudiana germplasm in liquid nitrogen.

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.

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).

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.

Increased sodium chloride (NaCl) salinity effects on bitter almond (Amygdalus communis L.) growth, cell osmolarity and nutrient acquisition were studied in vitro. Elevated salinity from 0.0 (control) to 50, 75, 100 mM NaCl resulted in reduction in shoot growth (shoot height, shoot dry weight) and rooting (rooting percentage, root number, root length). Reductions in growth parameters were accompanied with reductions in leaf cell sap osmolarity from −4.0 (control) to −11.1 [at 100 mM NaCl]. Acquisition of nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), and magnesium (Mg) was decreased with elevated salinity in the medium. On the other hand, micronutrients, zinc (Zn), copper (Cu), and manganese (Mn) as well as sodium (Na) increased with elevated salinity levels in the medium. Iron (Fe) was negatively affected and its acquisition was decreased with imposed salinity in the medium accompanied with increased chlorosis percentage. However, growth did not stop under the imposed treatments indicating a h...

Seeds of broomrape (Orobanche cernua) were exposed to 0, 25, 50, 75, and 100 mM NaCl solutions during their preconditioning period (14 days of moisture) under laboratory conditions and induced to germinate by synthetic germination stimulant (GR24). There was significant reduction in seed germination with increased salt concentration as shown in 35.2, 32.5, 23.6, 14.3, and 9.2% germination, respectively. Exposure of Orobanche cernua seeds to 0.0, 1.0, 1.25, and 1.5 M levels of NaCl for 9 hours resulted in 29.4, 21.3, 20.5, and 17.4% germination, respectively. Water preconditioned seeds showed Heavier protein profile bands of 6.5-14.2 KDa than those of dry seeds. Seeds treated with 0.75 M NaCl showed profile similar with that of water preconditioned ones, plus an extra band at 29-36 KDa. The protein profiles of 1.0 and 1.5 M NaCl treated seeds showed weaker bands with the absence of 29-36 KDa band.

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...

Greenhouse experiment was conducted to evaluate the effect of arbuscular mycorrhizal fungi (AMF) on plant growth, and nutrient uptake in saline soils with different salt and phosphorus (P) levels. The following treatments were included in this experiment: (i) Soil A, with salt level of 16.6 dS m−1 and P level of 8.4 mg kg−1; (ii) Soil B, with salt level of 6.2 dS m−1 and P level of 17.5 mg kg−1; and (iii) Soil C, with salt level of 2.4 dS m−1 and P level of 6.5 mg kg−1. Soils received no (control) or 25 mg P kg−1 soil as triple super phosphate and were either not inoculated (control) or inoculated with a mixture of AM (AM1) and/or with Glomus intraradices (AM2). All pots were amended with 125 mg N kg−1 soil as ammonium sulfate. Barley (Hordeum vulgar L., cv. “ACSAD 6”) was grown for five weeks. Plants grown on highly saline soils were severely affected where the dry weight was significantly lower than plants growing on moderately and low saline soils. The tiller number and the plant height were a...

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...