{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T08:10:11Z","timestamp":1783930211230,"version":"3.55.0"},"update-to":[{"DOI":"10.12688\/f1000research.173637.1","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T00:00:00Z","timestamp":1783900800000}},{"DOI":"10.12688\/f1000research.173637.2","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T00:00:00Z","timestamp":1783900800000}}],"reference-count":44,"publisher":"F1000 Research Ltd","license":[{"start":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T00:00:00Z","timestamp":1783900800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"self funding"}],"content-domain":{"domain":["f1000research.com"],"crossmark-restriction":false},"short-container-title":["F1000Res"],"abstract":"<ns5:p>\n                    Background Lavender is a crucial source of secondary metabolites with medicinal and industrial importance. Its exploitation faces challenges due to the limited availability of raw materials and their sensitivity to climate. Tissue culture techniques, particularly callus culture, represent the most sustainable method for the continuous production of these active compounds. Methods This study focused on developing an effective protocol for inducing and stimulating callus cultures in lavender plants. Callus formation was stimulated from leaf tissue has cultured on MS medium supplemented with different concentrations of 2, 4-D (0, 1, 2 and 3) mg L\n                    <ns5:sup>\u22121<\/ns5:sup>\n                    . The cultures were incubated under light or dark conditions. In the multiplication: calli (Cultured in MS medium containing 1 mg, 2,4-D incubated in the dark) was transferred to MS medium containing different 2,4-D (0, 1, 2 and 3 mg.L\n                    <ns5:sup>\u22121<\/ns5:sup>\n                    ) and BA (0,1 and 2 mg.L\n                    <ns5:sup>\u22121<\/ns5:sup>\n                    ). The multiplied callus (Cultured in MS medium containing 1mg L\n                    <ns5:sup>\u22121<\/ns5:sup>\n                    2, 4-D and 2 mg L\n                    <ns5:sup>\u22121<\/ns5:sup>\n                    BA) was transferred into two nutrient media, MS and B5, each supplemented with different levels of chitosan (0, 50, 100, and 200 mg.L\n                    <ns5:sup>\u22121<\/ns5:sup>\n                    ) along with combination of 1 mg.L\n                    <ns5:sup>\u22121<\/ns5:sup>\n                    2, 4-D and 2 mg.L\n                    <ns5:sup>\u22121<\/ns5:sup>\n                    BA. Results The results indicated that the optimal concentration for callus induction was 1 mg L\n                    <ns5:sup>\u22121<\/ns5:sup>\n                    of 2,4-D, with the highest callus formation rate 95% recorded under dark conditions. Among all treatments, the combination of 2, 4-D 1 mg L\n                    <ns5:sup>\u22121<\/ns5:sup>\n                    and BA 2 mg L\n                    <ns5:sup>\u22121<\/ns5:sup>\n                    was the most effective for sustainable callus growth. Regarding the nutrient medium, B5 was more efficient than the MS medium. For the biostimulants, Chitosan at 100 mg L\n                    <ns5:sup>\u22121<\/ns5:sup>\n                    recorded the highest efficiency in both fresh and dry callus weights. Conclusions This study underscores the necessity of integrating hormonal, nutritional and environmental factors for optimal callus production.\n                  <\/ns5:p>","DOI":"10.12688\/f1000research.173637.3","type":"journal-article","created":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T07:39:09Z","timestamp":1783928349000},"page":"407","update-policy":"https:\/\/doi.org\/10.12688\/f1000research.crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Evaluation of the efficiency of the optimal combination of\u00a0 hormonal, physical\u00a0 and biological stimuli in the induction and growth of callus in Lavandula angustifolia Mill"],"prefix":"10.12688","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1840-1681","authenticated-orcid":false,"given":"Mohammed Abdulgafor","family":"Mohammed","sequence":"first","affiliation":[{"name":"Department of Biotechnology Ramadi 31001 University of Anbar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masara","family":"Dafer Alani","sequence":"additional","affiliation":[{"name":"Department of Studies and Planning Ramadi 31001 University of Anbar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-4524-6106","authenticated-orcid":false,"given":"Ahmed O.","family":"Mashaan","sequence":"additional","affiliation":[{"name":"Department of Biotechnology Ramadi 31001 University of Anbar"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ali F.","family":"Almehemdi","sequence":"additional","affiliation":[{"name":"Center of Desert Studies, Department of conservation Agriculture Ramadi 31001 University of Anbar"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"2560","published-online":{"date-parts":[[2026,7,13]]},"reference":[{"key":"ref1","first-page":"1126-1130","article-title":"In vitro influences of sugar nutrition and light condition on accumulation of some phytochemicals in Verbascum thapsus L. culture.","volume":"20","author":"A Abed","year":"2020","journal-title":"Plant Archives."},{"key":"ref2","doi-asserted-by":"crossref","first-page":"14542","DOI":"10.15835\/nbha53314542","article-title":"Impact of media type on callus morphology and standardization of Withania somnifera cell culture for enhanced secondary metabolite production.","volume":"53","author":"M Adil","year":"2025","journal-title":"Notulae Botanicae Horti Agrobotanici Cluj-Napoca."},{"key":"ref3","doi-asserted-by":"crossref","first-page":"1113-1124","DOI":"10.54910\/sabrao2022.54.5.13","article-title":"In vitro production of active compounds in the callus of the chia plant (Salvia hispanica).","volume":"54","author":"A Al-Alwani","year":"2022","journal-title":"SABRAO Journal of Breeding and Genetics."},{"key":"ref4","doi-asserted-by":"publisher","DOI":"10.17605\/OSF.IO\/AMV4Z","article-title":"raw data for lavender study.","author":"A Almehemdi","year":"2025, November 26"},{"key":"ref5","first-page":"899-912","article-title":"Stimulation and production of some active compounds in the chia plant with melatonin in vitro.","volume":"20","author":"M Alrawi","year":"2022","journal-title":"Agron. 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Ethnopharmacol."}],"container-title":["F1000Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/f1000research.com\/articles\/15-407\/v3\/xml","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/f1000research.com\/articles\/15-407\/v3\/pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/f1000research.com\/articles\/15-407\/v3\/iparadigms","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T07:39:12Z","timestamp":1783928352000},"score":1,"resource":{"primary":{"URL":"https:\/\/f1000research.com\/articles\/15-407\/v3"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,13]]},"references-count":44,"URL":"https:\/\/doi.org\/10.12688\/f1000research.173637.3","relation":{"has-version":[{"id-type":"doi","id":"10.12688\/f1000research.173637.2","asserted-by":"subject"},{"id-type":"doi","id":"10.12688\/f1000research.173637.1","asserted-by":"subject"}]},"ISSN":["2046-1402"],"issn-type":[{"value":"2046-1402","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,13]]},"assertion":[{"value":"Indexed","URL":"https:\/\/f1000research.com\/articles\/15-407\/v3#article-reports","order":0,"name":"referee-status","label":"Referee status","group":{"name":"current-referee-status","label":"Current Referee Status"}},{"value":"10.5256\/f1000research.202213.r493247, Duha Mysire Majeed, Plant Biotechnology, Al-Nahrain University \/ Biotechnology Research center, Baghdad, 29 Jun 2026, version 2, indexed","URL":"https:\/\/f1000research.com\/articles\/15-407\/v2#referee-response-493247","order":0,"name":"referee-response-493247","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.202213.r493248, Tassanai Jaruwattanaphan, Kasetsart University, Bangkok, 06 Jul 2026, version 2, indexed","URL":"https:\/\/f1000research.com\/articles\/15-407\/v2#referee-response-493248","order":1,"name":"referee-response-493248","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"<b>MOHAMMED ABDULGAFOR<\/b>; \n<i>Posted: 08 Jul 2026<\/i>; We sincerely thank the reviewer for the careful re-evaluation of our revised manuscript and for the positive assessment of the improvements made. We greatly appreciate your recognition that the taxonomic inconsistency, Figure 1 scaling, chitosan characterization, clarification of GA\u2083 application, inclusion of the workflow diagram, and the revised terminology have substantially improved the quality and clarity of the manuscript.  We also thank you for pointing out the remaining issues that were unintentionally overlooked during the final revision.  Response to Comment 13 (Auxin mechanism): We have revised the Discussion to provide a more accurate explanation of the physiological mechanism underlying the reduced callus growth under light conditions. The statement has been reworded to avoid oversimplification, and an appropriate supporting reference has been added to strengthen this interpretation.  Response to Comment 14 (B5 medium superiority): We agree with the reviewer's suggestion. The Discussion has been expanded to explain that the superior performance of B5 medium is likely associated with differences in nutrient composition, particularly the nitrogen source ratio and vitamin formulation, compared with MS medium. Relevant supporting references have also been incorporated to support this explanation.  F-values and p-values in Tables 2\u20135: Thank you for requesting clarification. We confirm that the F-values and corresponding p-values have now been clearly presented in Tables 2\u20135 as indicated in our response letter. We carefully reviewed the final version to ensure that these statistical values are included and correctly displayed.  We sincerely appreciate your constructive comments and careful review. Your valuable suggestions have helped us further improve the scientific quality, clarity, and accuracy of the manuscript.","URL":"https:\/\/f1000research.com\/articles\/15-407\/v3#referee-comment-16613","order":2,"name":"referee-comment-16613","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"<b>MOHAMMED ABDULGAFOR<\/b>; \n<i>Posted: 08 Jul 2026<\/i>; We sincerely thank the reviewer for the careful re-evaluation of our revised manuscript and for the positive assessment of the improvements made. We greatly appreciate your recognition that the taxonomic inconsistency, Figure 1 scaling, chitosan characterization, clarification of GA\u2083 application, inclusion of the workflow diagram, and the revised terminology have substantially improved the quality and clarity of the manuscript.  We also thank you for pointing out the remaining issues that were unintentionally overlooked during the final revision.  Response to Comment 13 (Auxin mechanism): We have revised the Discussion to provide a more accurate explanation of the physiological mechanism underlying the reduced callus growth under light conditions. The statement has been reworded to avoid oversimplification, and an appropriate supporting reference has been added to strengthen this interpretation.  Response to Comment 14 (B5 medium superiority): We agree with the reviewer's suggestion. The Discussion has been expanded to explain that the superior performance of B5 medium is likely associated with differences in nutrient composition, particularly the nitrogen source ratio and vitamin formulation, compared with MS medium. Relevant supporting references have also been incorporated to support this explanation.  F-values and p-values in Tables 2\u20135: Thank you for requesting clarification. We confirm that the F-values and corresponding p-values have now been clearly presented in Tables 2\u20135 as indicated in our response letter. We carefully reviewed the final version to ensure that these statistical values are included and correctly displayed.  We sincerely appreciate your constructive comments and careful review. Your valuable suggestions have helped us further improve the scientific quality, clarity, and accuracy of the manuscript.","URL":"https:\/\/f1000research.com\/articles\/15-407\/v3#referee-comment-16614","order":3,"name":"referee-comment-16614","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.202213.r494919, Huda Enaya Mahood, University of Al-Qadisiyah, Al Diwaniyah, 01 Jul 2026, version 2, indexed","URL":"https:\/\/f1000research.com\/articles\/15-407\/v2#referee-response-494919","order":4,"name":"referee-response-494919","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"<b>MOHAMMED ABDULGAFOR<\/b>; \n<i>Posted: 05 Jul 2026<\/i>; \n<b>Response to Reviewer<\/b> We sincerely thank the reviewer for the careful evaluation of our manuscript and for the constructive comments, which have significantly improved its scientific quality. All comments were carefully considered, and the manuscript has been revised accordingly. Our point-by-point responses are presented below. \n<b>Comment 1:<\/b> \n<i>The introduction does not fully match the study title. The role of physical stimuli is not sufficiently discussed, despite being a key component of the research. Please include a brief background on physical stimuli and clearly align the study objective with the title.<\/i> \n<b>Response:<\/b> Thank you for this valuable suggestion. The Introduction has been revised to include a dedicated background describing the role of physical stimuli, particularly light and darkness, in regulating callus induction and growth through their influence on endogenous hormones, cellular dedifferentiation, and morphogenesis. In addition, the study objective has been rewritten to better reflect the title by emphasizing the integrated evaluation of hormonal, physical, and biological stimuli. \n<b>Comment 2:<\/b> \n<i>The details of the culture medium are lengthy and unnecessary.<\/i> \n<b>Response:<\/b> We agree with the reviewer. The description of culture medium preparation has been shortened by removing unnecessary procedural details while retaining all essential information required for reproducibility. \n<b>Comment 3:<\/b> \n<i>Sterilization: The phrase &quot;according to the method used by the researcher&quot; is inappropriate.<\/i> \n<b>Response:<\/b> Thank you for pointing this out. The sterilization procedure has been revised by removing this phrase and replacing it with a clear description of the protocol used, together with the appropriate reference. \n<b>Comment 4:<\/b> \n<i>The number of replicates and explants per replicate should be specified.<\/i> \n<b>Response:<\/b> This information has now been added to the Materials and Methods section. The number of biological replicates and the number of explants used in each replicate are clearly stated. \n<b>Comment 5:<\/b> \n<i>The Results section is weak because of the way the results are presented and discussed.<\/i> \n<b>Response:<\/b> We appreciate this important observation. The Results and Discussion have been extensively revised to improve clarity, organization, and scientific interpretation. The discussion now emphasizes the biological significance of the findings rather than simply describing the numerical results. \n<b>Comment 6:<\/b> \n<i>The numerical results are repeated; the discussion should focus on interpretation.<\/i> \n<b>Response:<\/b> We agree. Repetitive descriptions of numerical values have been removed, and the Discussion has been rewritten to focus on interpreting the results in relation to previous studies and the underlying physiological mechanisms. \n<b>Comment 7:<\/b> \n<i>The discussion of the effect of darkness is too brief.<\/i> \n<b>Response:<\/b> Thank you for this suggestion. The discussion concerning the influence of darkness has been substantially expanded. It now explains the physiological basis of darkness-induced callus formation, including its effects on auxin stability, cellular dedifferentiation, and morphogenic responses, and is supported by additional relevant literature. \n<b>Comment 8:<\/b> \n<i>The superiority of B5 over MS was not adequately discussed.<\/i> \n<b>Response:<\/b> We appreciate this comment. The Discussion has been expanded to explain the likely reasons for the superior performance of Gamborg's B5 medium, including differences in nitrogen sources, nitrate\/ammonium balance, vitamin composition, and their influence on cellular metabolism and biomass accumulation. Appropriate references have also been added. \n<b>Comment 9:<\/b> \n<i>The statement regarding reduced auxin effectiveness under light requires a reference or rewording.<\/i> \n<b>Response:<\/b> Thank you for identifying this issue. The statement has been revised and is now supported with appropriate references describing the influence of light on auxin metabolism and signaling. \n<b>Comment 10:<\/b> \n<i>The conclusion mainly summarizes the results.<\/i> \n<b>Response:<\/b> We agree with the reviewer. The Conclusion has been rewritten to emphasize the broader scientific significance of the study, highlighting the importance of integrating hormonal, physical, nutritional, and biological stimuli for optimizing callus production in \n<i>Lavandula angustifolia<\/i> and its potential applications in plant biotechnology and secondary metabolite production. Once again, we sincerely thank the reviewer for the insightful comments and valuable suggestions, which have substantially improved the quality, clarity, and scientific rigor of the manuscript.","URL":"https:\/\/f1000research.com\/articles\/15-407\/v3#referee-comment-16588","order":5,"name":"referee-comment-16588","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"The author(s) declared that no grants were involved in supporting this work","order":6,"name":"grant-information","label":"Grant Information"},{"value":"This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.","order":0,"name":"copyright-info","label":"Copyright"}]}}