RAPD for Assessment of Thymes Genetic Diversity in Palestine

Several thymes species were grown in Palestine and have socio-economical values. Traditionally, morphological features are used for their taxonomy and discrimination. However, these methods fail to provide an accurate for discrimination and authentication, suggesting the use of other method inevitable. In this study, RAPD method was used for genotypes identification and characterization of five most socio-economical Palestinian thymes: Thymus syriacus, Thymus fruticosus, Thymus incanus, Thymus majorana, and Thymus capitatus. Eight out of ten decamer primers were tested for their ability to generate polymorphism from selected thyme species using RAPD-PCR, and the obtained data were analyzed. The primers (OPD-19, OPH-02 and OPAN-08) were generated 78.6% average polymorphism across five studied Thyme species. Pairwise similarity analysis revealed banding patterns between the studied plant species ranged from 0.18 to 0.67. By this study, genetic diversities of studied five Palestinian Thyme species were ascertained successfully using RAPD markers, concluding that it could be useful tools for identifying Thymes species in any putative breeding programs that will be carried in the country

Biotechnology for conservation of palestinian medicinal plants

Many plants of Palestinian flora are facing the risk of endanger, due to agricultural practices, environmental threats and consumption changes. In the absence of National program to conserve the Palestinian heritage of plant diversity, a tentative research work aimed in trial usage of available biotechnology’s methods for conserving several popular plants of medical, cultural and economical importance's. Tissue culturing of anise (Pimipnella anisum), sage (Salvia palestina), fenugreek (Trigonella sps), wild peppermint (Mentha spicita L.) and akoub (Gandelia tournefortii); using MS-media with specific plant growth regulators were successfully applied. Protocols for enhancing callus culturing, organogenesis and micropropagation of these tentatively threatened wild plants were developed and optimized in this research work. Based on the successfulness of propagation in vitro of these plants, a call for establishment of a Palestinian germplasm collection to conserve the Flora Palestina had been reported.This research work was partially funded by Arab American University and Al-Quds University. The authors are thankful to Layth Sbeihat, Shatela Jaradat, Kholod Abu Al-rub and Yusuf Modallal for their contribution to parts of this research study

Adsorption of Heavy Metals by Reed (Phragmites australis) as a Potential Clean Water Technology

In this study the adsorption behavior of reed tissues has been investigated in a batch system in order to determine its applicability in treating wastewater. The optimum conditions for the treatment process were investigated by observing the influence of organ of plant (leaf, stem, root), pH levels, the presence of competing ions, initial concentration and of the reed tissues. The results indicated that the adsorption capacity was strongly affected by the initial concentration, pH levels and organ of plant. Optimum adsorption of Pb (II) was observed at pH 7.0, initial Pb(II) concentration 20 mg/L, temperature 25-45 0C and contact time 30 minute. It has a good fit for the Freundlich compared with Langmuir isotherm models. As well as, Pseudo second – order kinetic model was fit better than pseudo first order model. Phragmites Australia shows that it is feasible for this plant material as a novel adsorbent for Pb(II) removal in the future. Key words: Reed tissues, Pb(II), Adsorption,  Freundlich, Langmuir; Stem.

Molecular analysis of a California strain of Rupestris stem pitting-associated virus isolated from declining Syrah grapevines

The sequence of the genome of a Rupestris stem pitting-associated virus (RSPaV) isolated from a declining Syrah grapevine in California, designated the Syrah strain (RSPaV-SY) was determined. The genome of this strain had an overall nucleotide identity os 77% in comparison with RSPaV sequences in GenBank; the coat protein was the most conservd gene among RSPaV sequences among replicase was the least conserved gene. Phylogenetic analysis of partial coat protein and replicase gene sequences showed RSPaV-SY clustrd independently from the majority of RSPaV isolates

Real-time reverse transcription polymerase chain reaction development for rapid detection of Tomato brown rugose fruit virus and comparison with other techniques

[EN] Background: Tomato brown rugose fruit virus (ToBRFV) is a highly infectious tobamovirus that causes severe disease in tomato (Solanum lycopersicum L.) crops. In Italy, the first ToBRFV outbreak occurred in 2018 in several provinces of the Sicily region. ToBRFV outbreak represents a serious threat for tomato crops in Italy and the Mediterranean Basin. Methods: Molecular and biological characterisation of the Sicilian ToBRFV ToB-SIC01/19 isolate was performed, and a sensitive and specific Real-time RT-PCR TaqMan minor groove binder probe method was developed to detect ToBRFV in infected plants and seeds. Moreover, four different sample preparation procedures (immunocapture, total RNA extraction, direct crude extract and leaf-disk crude extract) were evaluated. Results: The Sicilian isolate ToB-SIC01/19 (6,391 nt) showed a strong sequence identity with the isolates TBRFV-P12-3H and TBRFV-P12-3G from Germany, Tom 1-Jo from Jordan and TBRFV-IL from Israel. The ToB-SIC01/19 isolate was successfully transmitted by mechanical inoculations in S. lycopersicum L. and Capsicum annuum L., but no transmission occurred in S. melongena L. The developed real-time RT-PCR, based on the use of a primer set designed on conserved sequences in the open reading frames3, enabled a reliable quantitative detection. This method allowed clear discrimination of ToBRFV from other viruses belonging to the genus Tobamovirus, minimising false-negative results. Using immunocapture and total RNA extraction procedures, the real-time RT-PCR and end-point RT-PCR gave the same comparable results. Using direct crude extracts and leaf-disk crude extracts, the end-point RT-PCR was unable to provide a reliable result. This developed highly specific and sensitive real-time RT-PCR assay will be a particularly valuable tool for early ToBRFV diagnosis, optimising procedures in terms of costs and time.Panno, S.; Ruiz-Ruiz, S.; Giovani Carusso, A.; Alfaro Fernández, AO.; Font San Ambrosio, MI.; Davino, S. (2019). Real-time reverse transcription polymerase chain reaction development for rapid detection of Tomato brown rugose fruit virus and comparison with other techniques. PeerJ. 7:1-20. https://doi.org/10.7717/peerj.7928S1207Alkowni, R., Alabdallah, O., & Fadda, Z. (2019). Molecular identification of tomato brown rugose fruit virus in tomato in Palestine. Journal of Plant Pathology, 101(3), 719-723. doi:10.1007/s42161-019-00240-7Cambrón-Crisantos, J. M., Rodríguez-Mendoza, J., Valencia-Luna, J. B., Alcasio-Rangel, S., García-Ávila, C. D. J., López-Buenfil, J. A., & Ochoa-Martínez, D. L. (2018). Primer reporte de Tomato brown rugose fruit virus (ToBRFV) en Michoacán, México. Revista Mexicana de Fitopatología, Mexican Journal of Phytopathology, 37(1). doi:10.18781/r.mex.fit.1810-5Davino, S., Miozzi, L., Panno, S., Rubio, L., Davino, M., & Accotto, G. P. (2012). Recombination profiles between Tomato yellow leaf curl virus and Tomato yellow leaf curl Sardinia virus in laboratory and field conditions: evolutionary and taxonomic implications. Journal of General Virology, 93(12), 2712-2717. doi:10.1099/vir.0.045773-0Davino, S., Napoli, C., Dellacroce, C., Miozzi, L., Noris, E., Davino, M., & Accotto, G. P. (2009). Two new natural begomovirus recombinants associated with the tomato yellow leaf curl disease co-exist with parental viruses in tomato epidemics in Italy. Virus Research, 143(1), 15-23. doi:10.1016/j.virusres.2009.03.001Davino, S., Panno, S., Iacono, G., Sabatino, L., D’Anna, F., Iapichino, G., … Davino, M. (2016). Genetic variation and evolutionary analysis ofPepino mosaic virusin Sicily: insights into the dispersion and epidemiology. Plant Pathology, 66(3), 368-375. doi:10.1111/ppa.12582Ferriol, I., Rangel, E. A., Panno, S., Davino, S., Han, C. G., Olmos, A., & Rubio, L. (2015). Rapid detection and discrimination of fabaviruses by flow-through hybridisation with genus- and species-specific riboprobes. Annals of Applied Biology, 167(1), 26-35. doi:10.1111/aab.12204Fidan, H., Sarikaya, P., & Calis, O. (2019). First report of Tomato brown rugose fruit virus on tomato in Turkey. New Disease Reports, 39(1), 18-18. doi:10.5197/j.2044-0588.2019.039.018Hanssen, I. M., Lapidot, M., & Thomma, B. P. H. J. (2010). Emerging Viral Diseases of Tomato Crops. Molecular Plant-Microbe Interactions®, 23(5), 539-548. doi:10.1094/mpmi-23-5-0539Jacobi, V., Bachand, G. ., Hamelin, R. ., & Castello, J. . (1998). Development of a multiplex immunocapture RT-PCR assay for detection and differentiation of tomato and tobacco mosaic tobamoviruses. Journal of Virological Methods, 74(2), 167-178. doi:10.1016/s0166-0934(98)00086-xLarkin, M. A., Blackshields, G., Brown, N. P., Chenna, R., McGettigan, P. A., McWilliam, H., … Higgins, D. G. (2007). Clustal W and Clustal X version 2.0. Bioinformatics, 23(21), 2947-2948. doi:10.1093/bioinformatics/btm404Levitzky, N., Smith, E., Lachman, O., Luria, N., Mizrahi, Y., Bakelman, H., … Dombrovsky, A. (2019). The bumblebee Bombus terrestris carries a primary inoculum of Tomato brown rugose fruit virus contributing to disease spread in tomatoes. PLOS ONE, 14(1), e0210871. doi:10.1371/journal.pone.0210871Ling, K.-S., Tian, T., Gurung, S., Salati, R., & Gilliard, A. (2019). First Report of Tomato Brown Rugose Fruit Virus Infecting Greenhouse Tomato in the United States. Plant Disease, 103(6), 1439. doi:10.1094/pdis-11-18-1959-pdnLuria, N., Smith, E., Reingold, V., Bekelman, I., Lapidot, M., Levin, I., … Dombrovsky, A. (2017). A New Israeli Tobamovirus Isolate Infects Tomato Plants Harboring Tm-22 Resistance Genes. PLOS ONE, 12(1), e0170429. doi:10.1371/journal.pone.0170429Menzel, W., Knierim, D., Winter, S., Hamacher, J., & Heupel, M. (2019). First report of Tomato brown rugose fruit virus infecting tomato in Germany. New Disease Reports, 39(1), 1-1. doi:10.5197/j.2044-0588.2019.039.001Panno, S., Caruso, A. G., & Davino, S. (2017). The nucleotide sequence of a recombinant tomato yellow leaf curl virus strain frequently detected in Sicily isolated from tomato plants carrying the Ty-1 resistance gene. Archives of Virology, 163(3), 795-797. doi:10.1007/s00705-017-3674-9Panno, S., Caruso, A. G., & Davino, S. (2019). First Report of Tomato Brown Rugose Fruit Virus on Tomato Crops in Italy. Plant Disease, 103(6), 1443-1443. doi:10.1094/pdis-12-18-2254-pdnPanno, S., Caruso, A. G., Troiano, E., Luigi, M., Manglli, A., Vatrano, T., … Davino, S. (2019). Emergence of tomato leaf curl New Delhi virus in Italy: estimation of incidence and genetic diversity. Plant Pathology, 68(3), 601-608. doi:10.1111/ppa.12978Panno, S., Davino, S., Rubio, L., Rangel, E., Davino, M., García-Hernández, J., & Olmos, A. (2012). Simultaneous detection of the seven main tomato-infecting RNA viruses by two multiplex reverse transcription polymerase chain reactions. Journal of Virological Methods, 186(1-2), 152-156. doi:10.1016/j.jviromet.2012.08.003Panno, S., Ferriol, I., Rangel, E. A., Olmos, A., Han, C.-G., Martinelli, F., … Davino, S. (2014). Detection and identification of Fabavirus species by one-step RT-PCR and multiplex RT-PCR. Journal of Virological Methods, 197, 77-82. doi:10.1016/j.jviromet.2013.12.002Pelham, J. (1966). Resistance in tomato to tobacco mosaic virus. Euphytica, 15(2), 258-267. doi:10.1007/bf00022331Puchades, A. V., Carpino, C., Alfaro-Fernandez, A., Font-San-Ambrosio, M. I., Davino, S., Guerri, J., … Galipienso, L. (2017). Detection of Southern tomato virus by molecular hybridisation. Annals of Applied Biology, 171(2), 172-178. doi:10.1111/aab.12367Salem, N., Mansour, A., Ciuffo, M., Falk, B. W., & Turina, M. (2015). A new tobamovirus infecting tomato crops in Jordan. Archives of Virology, 161(2), 503-506. doi:10.1007/s00705-015-2677-

Candida Endophthalmiltis Following Penetrating Keratoplasty in Patient with Negative Donor Rim; A case report with review literature of diagnosis and treatment of fungal endophthalmitis.

 This is a case of young patient presented with granulomatous anterior and posterior uveitis, which turned to be fungal endophthalmitis after penetrating keratoplasty. Her symptoms were undetected because she was on systemic and topical steroids. Key words: Candida, Endophthalmiltis, Penetrating Keratoplasty, negative donner rim. The patient is 25 years old Caucasian female patient, previously medically free, who visited our department in the city of Amman, Jordan, for left penetrating keratoplasty for severe keratoconus. After an initial improvement in her vision and a smooth postoperative course, she presented with drop of vision, photophobia, and non-specific eye pain, on examination was found to have anterior granulomatous uveitis. She was started on systemic steroids and the topical steroids were increased in intensity. The initial systemic workup for granulomatous anterior uveitis was negative. However, culture from the aqueous was positive for Candida Galibrata, but the donor rim was negative. Later on the patient developed vitritis despite being on systemic fluconazole and topical Amphotericin B. She was treated with intravitreal Amphotericin B. The vitritis improved, but vitreous opacities developed which deteriorated her vision. A parsplana vitetrectony was done. Her final visual acuity remained poor because of opacified graft.    The patient’s unfortunate case represents a Candida endophthalmitis after penetrating keratoplasty despite being medically free