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Medicinal plants of Jordan are under threat due to several factors of habitat perturbation. Some of these factors include ruinous, over-harvesting, climate change, competition and invasion. In this study simple we employed a reliable method for micropropagation and assessment of antimicrobial activity of some important medicinal plants of Jordan. Seeds were surface sterilized and germinated on agar water media, and then shoots were cultured on Murashige and Skoog (MS) medium with the addition of 3% sucrose. To generate enough plant material, microshoots were transferred after fourteen days onto hormone free MS medium. Among all studied plants we found that Achellia millefolium and Moring perigrina plants were successful multiply in vitro containing different 6-benzyl amino purine (BA). For potential antimicrobial activity ex vitro (field leaf plants) and in vitro (plantlet) extracts were evaluated against some bacteria strain using ethanolic extracts. Both ex vitro and in vitro plants extract showed antimicrobial activity against the microorganism tested. The results from the study suggest that these two plants showed good antimicrobial activity against the different tested bacteria.

Background The presence of phenolic acids in edible products for human consumption is considered in relation to the production of odorant substances, with a variety of different aromas. Objective Phenolic substances anthocyanidins, anthocyanins, flavanols, flavones and isoflavones, flavones, flavonols, etc. – are extremely interesting as flavor additives, anti-aging or maturing agents, and color and aroma enhancers. Method The connection between flavoring properties on one hand and the presence of phenolic compounds on the other can be discussed in terms of food acceptance by consumers, especially with relation to the “Mediterranean Diet” lifestyle. Results The health perspectives of these and other food products related to Mediterranean Diet should be evaluated in the geographical ambit of the Mediterranean Basin, including several particular food and vegetable preparations – herbs and medicinal plants - of the Middle East. Conclusions The aim of this paper is to give a presentation on these specialties in relation to Jordan. Highlights Medicinal herbs have interesting health properties against digestive problems, parasitic worms, liver diseases, diabetes, skin problems, nervous, cardiocirculatory, and respiratory diseases.

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.

Moringa peregrina (Forssk) Fiori is one of the known medicinal plants in Jordan. It is used in traditional medicine to treat rheumatism and infections. Plant parts are used in the indigenous systems of human medicine for the treatment of a variety of ailments. Therefore, this study was conducted to investigate factors affecting M. peregrina in vitro propagation and its antimicrobial activity. Microshoots with apical meristem (10 or 15 mm in length) were subculture on MS medium supplemented with different concentration of BAP, Kinetin, Zeatin and Thidiazuron (TDZ). Maximum number of new shoots/explant (4.39) was obtained on Murashige and Skoog agar medium supplemented with 1.6 mg/L Zeatin. While at 0.4 mg/L BAP maximum shoot length was obtained (37.78 mm). Antibacterial activity of aqueous, methanolic, ethanolic, and acetonic extracts of both in vitro plantlets and ex vitro (field grown) M. peregrina were evaluated by the agar well-diffusion method against Klebsiella oxytoca ATCC 18182, Salmonella typhimurium ATCC 19430, Methicillin resistant, Staphcoccus aureus ATCC 29974, Klebsiella pneumonia ATCC13883, Proteus vulgaris ATCC 13315, Proteus mirabilis ATCC 12453, Enterobacter aerogenes ATCC 35029, Pseudomonas aeruginosa ATCC 27253, Escherichia coli O157:H7, Staphylococcus aureus ATCC 25923, Salmonella paratyphi ATCC 13076 and Escherichia coli ATCC 29522. The obtained results showed that ethanolic, acetonic and aqueous extract revealed a wide antibacterial activity. Ethanolic extract of in vitro M. peregrina plantlets showed the maximum inhibition zone against Staphcoccus aureus .

□ 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.

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.

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.

Aqueous extracts of nine plants, known to have medicinal activity, were tested for their toxicity against the sweet potato whitefly, Bemisia tabaci Genn. (Homoptera: Aleurodidae) compared to the toxicity of the insecticide, Imidacloprid. Extracts of Lepidiuim sativum L. (Brassicales: Brassicaceae) killed 71 % of early stage nymphs, which was not significantly different from mortality caused by Imidacloprid. Treatment of pupae with three plant extracts, L. sativum, Achillea biebersteinii L. (Asterales: Asteraceae), or Retama raetam (Forssk.) Webb and Berthel (Fabales: Fabaceae) prevented adult development, and treatment with R. raetam extract killed adults, at levels that were not significantly different from Imidacloprid. None of the other plants showed significant toxicity. However extracts of four plants, Pimpinella anisum L. (Apiales: Apiaceae), Galium longifolium (Sibth. and SM.) (Gentianales: Rubiaceae), R. raetam and Ballota undulata Bentham (Lamiales: Lamiaceae) had a repellent effect.

Plantlets derived from in vitro culture might exhibit somoclonal variation which is often heritable, and molecular variations may be generated in vitro. Since the direction of most studies is toward nuclear genome, there is a little known about the DNA of organelles. This study was conducted to test the genetic stability of wheat organelles genomes for plantlets produced by anther culture using restriction fragment length polymorphism (RFLP) analyses. One of the intergenic regions of cpDNA and one of mtDNA introns were amplified with polymerase chain reaction (PCR). The PCR products were then sequenced and digested with four restriction endonucleases (EcoR1, BamH1, NdeI and Sac1). The amplified product from cpDNA was 1000 bp in size, and digested only with NdeI into two bands with 650 and 350 bp. The amplified product from mtDNA was 1550 bp in size, and digested only with Sac1 into two bands with 1220 and 330 bp. The results obtained showed that no noticeable difference can be detected between doubled haploid plantlets and parental plants at the level of ctDNA and mtDNA organization. It can be concluded that in vitro culture by itself does not systematically generate a cytoplasmic variation in plant cells.Key words: RFLP analysis, wheat plantlets, wheat anther culture, doubled haploids, genetic stability, mitochondria and chloroplast genome.

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.

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 ...

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.

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.

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.

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.

Superoxide dismutase is considered to be a crucial component in biological defense against oxidative stress. A full-length cDNA clone encoding a putative chloroplastic Cu/Zn-SOD from barley seedlings ( Hordeum vulgar cv. Rum) was isolated by RT-PCR from cDNA synthesized from leaf mRNA. HvSOD cDNA is 760 bp in length, which translates an ORF of 201 amino acids that correspond to a protein of predicted molecular mass of 20.21 kDa and a pI of 5.31.The deduced amino acid sequence of HvSOD showed higher identity (72–95%) with the sequences of the chloroplastic Cu/Zn-SODs than those of the cytosol from other species. Moreover the cloned sequence showed all the typical structural motifs of plant chloroplastic SODs with a 47 bp plastid targeting peptide. These results suggested that the HvSOD is chloroplastic. Reverse transcription-PCR analysis was used here to investigate the expression patterns of HvSOD in response to abiotic stresses. The HvSOD gene was induced in leaf tissues in response to drought, cold stress and exposure to stress related chemicals, indicating that it is involved in the antioxidative process under various environmental stresses.