{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T03:24:22Z","timestamp":1789183462020,"version":"build-2803163510"},"reference-count":30,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,9,21]],"date-time":"2022-09-21T00:00:00Z","timestamp":1663718400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Plant Sci."],"abstract":"<jats:p>\n                    <jats:italic>In vitro<\/jats:italic>\n                    plant cell and tissue cultures are potent tools to propagating germplasm resources in conserving and managing plant genetic resources. A reliable micropropagation protocol was developed for efficient callus proliferation and direct and indirect shoot regeneration of Meseika (\n                    <jats:italic>Haplophyllum tuberculatum<\/jats:italic>\n                    ). With the applied sterilization procedure, immature, unopened\n                    <jats:italic>H. tuberculatum<\/jats:italic>\n                    seed pods can be identified as a potent explant with high viability and low contamination percentage. Multiple shoots were regenerated from leaf and stem explants through direct organogenesis on Murashige and Skoog\u2019s (MS)\u2009+\u20093% sucrose medium amended with BAP. Indirect regeneration of several shoots was achieved on 1\/2 MS\u2009+\u20091% sucrose media amended with 2 and 4\u2009mg\/l BAP. An efficient callus proliferation from both explants can be achieved by supplementing the MS media with NAA and BAP. All the cultures were incubated in a controlled growth chamber under 5\/19\u2009h light\/dark photoperiod, temperature (25\u2009\u00b1\u20092\u00b0C), and 60% relative humidity (RH).10 ISSR (Inter Simple Sequence Repeat) markers were screened to test the genetic fidelity of regenerated\n                    <jats:italic>H. tuberculatum<\/jats:italic>\n                    shoots. Callus development was observed after 15\u2009days and shoot regeneration was occurred after 30\u2009days after callus initiation. 10 ISSR primers produced a total of 39 clear, distinct amplicons. 75, 60, 40, and 16% polymorphism percentages were recorded by the ISSR primer 11, 7, 5, and 4, respectively. The developed micropropagation protocol is appropriate for rapid\n                    <jats:italic>in-vitro<\/jats:italic>\n                    multiplication of\n                    <jats:italic>H. tuberculatum<\/jats:italic>\n                    shoots and callus.\n                  <\/jats:p>","DOI":"10.3389\/fpls.2022.995825","type":"journal-article","created":{"date-parts":[[2022,9,21]],"date-time":"2022-09-21T10:32:20Z","timestamp":1663756340000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":13,"title":["Efficient shoot regeneration of medicinal plant Haplophyllum tuberculatum by direct and indirect organogenesis and genetic fidelity assessment using Inter Simple Sequence Repeats markers"],"prefix":"10.3389","volume":"13","author":[{"given":"Mohammed","family":"Alsafran","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kokila","family":"Wickramanayake","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kamal","family":"Usman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Talaat","family":"Ahmed","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2022,9,21]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"635","DOI":"10.21608\/zjar.2019.40947","article-title":"Some factors affecting micropropagation of Haplophyllum tuberculatum L. Zagazig","volume":"46","author":"Abdallah","year":"2019","journal-title":"J. Agric. Res."},{"key":"ref2","doi-asserted-by":"publisher","first-page":"12","DOI":"10.55493\/5003.v12i1.4449","article-title":"Callus induction of fenugreek trigonella foenum-graecum via auxin combined with cytokinins hormones, and assessment of toxicity via brine shrimp assay","volume":"12","author":"Agha","year":"2022","journal-title":"J. Asian Sci. Res."},{"key":"ref3","doi-asserted-by":"publisher","first-page":"942","DOI":"10.1080\/13102818.2016.1199287","article-title":"Use of ISSR markers to assess the genetic diversity in wild medicinal Ziziphus spina-christi (L.) Willd. Collected from different regions of Saudi Arabia","volume":"30","author":"Alansi","year":"2016","journal-title":"Biotechnol. Biotechnol. Equip."},{"key":"ref4","doi-asserted-by":"publisher","first-page":"107","DOI":"10.1016\/j.jep.2004.08.039","article-title":"Chemical composition, antibacterial and antifungal activities of the essential oil of Haplophyllum tuberculatum from Oman","volume":"96","author":"Al-Burtamani","year":"2005","journal-title":"J. Ethnopharmacol."},{"key":"ref5","doi-asserted-by":"publisher","DOI":"10.4066\/2591-7897-C1-002","article-title":"In vitro propagation of native plant species from Kuwait Desert","author":"Almazrooei","year":"2018","journal-title":"J. Agric. Sci. Bot."},{"key":"ref6","doi-asserted-by":"publisher","first-page":"101","DOI":"10.1016\/j.ejbas.2017.04.003","article-title":"Evaluation of antioxidant and cytotoxic activities of different extracts of folk medicinal plant Hapllophyllum tuberculatum","volume":"4","author":"Al-Muniri","year":"2017","journal-title":"Egypt. J. Bas. Appl. Sci."},{"key":"ref7","doi-asserted-by":"publisher","first-page":"52","DOI":"10.1016\/j.jobab.2021.09.004","article-title":"Characterization of antioxidant and antibacterial compounds from aerial parts of Haplophyllum tuberculatum","volume":"7","author":"Al-Saeghi","year":"2022","journal-title":"J. Bioresour. Bioprod."},{"key":"ref8","doi-asserted-by":"publisher","first-page":"e20210017","DOI":"10.1590\/2179-8087-FLORAM-2021-0017","article-title":"Indirect shoot regeneration using 2,4-D induces somaclonal variations in Cinchona officinalis","volume":"28","author":"Armijos-Gonz\u00e1lez","year":"2021","journal-title":"Floresta Ambient."},{"key":"ref9","doi-asserted-by":"publisher","first-page":"393","DOI":"10.4038\/jnsfsr.v49i3.10305","article-title":"In vitro culture of Nymphaea nouchali seeds; a conservation approach for a vulnerable species","volume":"49","author":"Attanayaka","year":"2021","journal-title":"J. Natl. Sci. Found."},{"key":"ref10","doi-asserted-by":"publisher","first-page":"e00450","DOI":"10.1016\/j.btre.2020.e00450","article-title":"Plant tissue culture as a perpetual source for production of industrially important bioactive compounds","volume":"26","author":"Chandran","year":"2020","journal-title":"Biotechnol. Rep."},{"key":"ref11","doi-asserted-by":"publisher","first-page":"1436","DOI":"10.3390\/plants9111436","article-title":"The establishment of an efficient callus induction system for lotus (Nelumbo nucifera)","volume":"9","author":"Deng","year":"2020","journal-title":"Plants"},{"key":"ref12","doi-asserted-by":"publisher","first-page":"151","DOI":"10.3109\/13880209.2013.819517","article-title":"Biological activity of HPLC-characterized ethanol extract from the aerial parts of Haplophyllum tuberculatum","volume":"52","author":"Eissa","year":"2014","journal-title":"Pharm. Biol."},{"key":"ref13","first-page":"127","author":"Elmaghabi","year":"2017"},{"key":"ref023","volume-title":"Haplophyllum tuberculatum \u2013 Flora of Qatar","year":"2021"},{"key":"ref15","doi-asserted-by":"publisher","first-page":"249","DOI":"10.1007\/978-1-0716-0997-2_14","article-title":"Inter-simple sequence repeats (ISSR), microsatellite-primed genomic profiling using universal primers","volume":"2222","author":"Gemmill","year":"2021","journal-title":"Methods Mol. Biol."},{"key":"ref16","doi-asserted-by":"publisher","first-page":"282","DOI":"10.3389\/fpls.2019.00282","article-title":"Modeling and optimizing in vitro sterilization of chrysanthemum via multilayer perceptron-non-dominated sorting genetic algorithm-II (MLP-NSGAII)","volume":"10","author":"Hesami","year":"2019","journal-title":"Front. Plant Sci."},{"key":"ref17","doi-asserted-by":"publisher","first-page":"54","DOI":"10.1007\/s13205-016-0389-7","article-title":"Somaclonal variations and their applications in horticultural crops improvement","volume":"6","author":"Krishna","year":"2016","journal-title":"3 Biotech"},{"key":"ref18","doi-asserted-by":"publisher","first-page":"446","DOI":"10.1007\/s42535-020-00128-3","article-title":"In vitro micropropagation and genetic fidelity studies using SCoT and ISSR primers in Annona reticulata L.: an important medicinal plant","volume":"33","author":"Kudikala","year":"2020","journal-title":"Vegetos"},{"key":"ref19","doi-asserted-by":"publisher","first-page":"964","DOI":"10.4236\/ajps.2013.45119","article-title":"Molecular analysis of genetic fidelity in micropropagated plants of Stevia rebaudiana Bert. Using ISSR marker","volume":"4","author":"Lata","year":"2013","journal-title":"Am. J. Plant Sci."},{"key":"ref01","first-page":"62","article-title":"Zur samenkeimung mediterraner ophrys","volume":"22","author":"Lucke","year":"1971","journal-title":"Die Orchidee"},{"key":"ref03","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1111\/j.1399-3054.1962.tb08052.x","article-title":"A revised medium for rapid growth and bio assays with tobacco tissue cultures","volume":"15","author":"Murashige","year":"1962","journal-title":"Plant Physiol."},{"key":"ref20","doi-asserted-by":"publisher","first-page":"480","DOI":"10.1007\/s11627-008-9143-0","article-title":"The effect of auxin type and cytokinin concentration on callus induction and plant regeneration frequency from immature inflorescence segments of seashore Paspalum (Paspalum vaginatum Swartz)","volume":"44","author":"Neibaur","year":"2008","journal-title":"In Vitro Cell. Dev. Biol."},{"key":"ref02","author":"Norton","year":"2009","journal-title":"An Illustrated Checklist of the Flora of Qatar"},{"key":"ref21","doi-asserted-by":"publisher","first-page":"58","DOI":"10.14295\/cs.v9i1.2034","article-title":"Concentrations of 6-benzylaminopurine (BAP) in micropropagation of banana \u2018Farta Velhaco\u2019 (AAB)","volume":"9","author":"Pereira","year":"2018","journal-title":"Comun. Sci."},{"key":"ref22","first-page":"125","article-title":"Haplophyllum tuberculatum: an overview","volume":"5","author":"Raissi","year":"2016","journal-title":"J. Herbmed. Pharmacol."},{"key":"ref23","volume-title":"Managing Global Genetic Resources: Agricultural Crop Issues and Policies","year":"1993"},{"key":"ref24","doi-asserted-by":"publisher","first-page":"60","DOI":"10.21786\/bbrc\/11.1\/9","article-title":"Plant regeneration from direct and indirect organogenesis and assessment of genetic fidelity in Saccharum officinarum using DNA-based markers. Bioscience biotechnology research","volume":"11","author":"Thorat","year":"2018","journal-title":"Communications"},{"key":"ref25","doi-asserted-by":"publisher","first-page":"6649","DOI":"10.1007\/s00253-021-11539-w","article-title":"Production of bioactive plant secondary metabolites through in vitro technologies\u2014status and outlook","volume":"105","author":"Wawrosch","year":"2021","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref26","doi-asserted-by":"publisher","first-page":"265","DOI":"10.1016\/j.fitote.2013.06.009","article-title":"Evaluation of genetic fidelity among micropropagated plants of Gloriosa superba L. using DNA-based markers \u2014 a potential medicinal plant","volume":"89","author":"Yadav","year":"2013","journal-title":"Fitoterapia"},{"key":"ref27","doi-asserted-by":"publisher","first-page":"2367","DOI":"10.4314\/ajb.v9i16","article-title":"Optimization of callus induction and plant regeneration from germinating seeds of sweet sorghum (Sorghum bicolor Moench)","volume":"9","author":"Zhao","year":"2010","journal-title":"Afr. J. Biotechnol."}],"container-title":["Frontiers in Plant Science"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fpls.2022.995825\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,10,3]],"date-time":"2022-10-03T07:01:43Z","timestamp":1664780503000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fpls.2022.995825\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,9,21]]},"references-count":30,"alternative-id":["10.3389\/fpls.2022.995825"],"URL":"https:\/\/doi.org\/10.3389\/fpls.2022.995825","relation":{},"ISSN":["1664-462X"],"issn-type":[{"value":"1664-462X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,9,21]]},"article-number":"995825"}}