{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,29]],"date-time":"2026-06-29T03:46:00Z","timestamp":1782704760736,"version":"3.54.5"},"reference-count":47,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T00:00:00Z","timestamp":1782432000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key R&D Program of China","doi-asserted-by":"publisher","award":["2024YFD2200600"],"award-info":[{"award-number":["2024YFD2200600"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Plants"],"abstract":"<jats:p>Ziziphus mauritiana Lam. (ber or Indian jujube) is a stress-tolerant dryland fruit tree valued for its nutritious fruit and ability to grow on marginal land. However, the phenotypic, cytotypic and genetic structure of its wild-growing\/naturalized germplasm in Pakistan remains poorly characterized. This study provides an integrated assessment of phenotypic, cytotypic and simple-sequence-repeat (SSR) diversity in 100 wild-growing\/naturalized accessions collected from Khyber Pakhtunkhwa, Punjab and Sindh to establish a baseline for conservation and germplasm management. We recorded 37 morphological and biochemical traits, estimated ploidy levels by flow cytometry (using diploid Z. jujuba \u2018Dongzao\u2019 as a reference), and genotyped a representative subset of 60 accessions with 14 SSR markers scored as a binary presence\/absence matrix. Substantial phenotypic variation was observed, especially in canopy architecture, leaf traits and stone-related characteristics; fruit quality traits (total soluble solids, vitamin C, and acidity) varied within a narrower range. Province explained only a modest proportion of phenotypic variation (PERMANOVA R2 = 0.059\u20130.109; p &lt; 0.01), with extensive overlap among regions. Flow cytometry revealed polyploid diversity: hexaploid (2n = 6x = 72) accessions dominated (46.7%), followed by octoploid (2n = 8x = 96; 31.7%) and tetraploid (2n = 4x = 48; 21.7%) cytotypes. SSR analysis showed moderate within-province diversity (Nei\u2019s H \u2248 0.51) but negligible genetic differentiation among provinces (R2 = 0.030; p = 0.60; \u03a6 \u2248 \u22120.011), indicating weak geographic structuring. Wild-growing\/naturalized Z. mauritiana in Pakistan forms a diverse, weakly structured gene pool in which most variation occurs within rather than among provinces. Sampling for conservation and germplasm management should, therefore, prioritize phenotypic distinctiveness, cytotype representation and ecological context rather than geographic origin alone. Experimental validation of any adaptive or agronomic advantages of particular cytotypes is needed before breeding recommendations can be made.<\/jats:p>","DOI":"10.3390\/plants15131974","type":"journal-article","created":{"date-parts":[[2026,6,29]],"date-time":"2026-06-29T02:16:44Z","timestamp":1782699404000},"page":"1974","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Integrated Phenotypic, Cytotypic, and Microsatellite Diversity Analysis of Wild-Growing\/Naturalized Ber (Ziziphus mauritiana Lam.) Across Pakistan: Implications for Germplasm Conservation and Breeding"],"prefix":"10.3390","volume":"15","author":[{"given":"Mian Fazli","family":"Basit","sequence":"first","affiliation":[{"name":"Key Laboratory of National Forestry and Grassland Administration on Silviculture in Loess Plateau, College of Forestry, Northwest A&F University, No. 3 Taicheng Road, Yangling 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nadeem","family":"Bhanbhro","sequence":"additional","affiliation":[{"name":"Sanya Institute, Hainan Academy of Agricultural Sciences, Sanya 572000, China"},{"name":"Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China"},{"name":"National Nanfan Research Institute, Chinese Academy of Agricultural Sciences, Sanya 572000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fazli","family":"Rahim","sequence":"additional","affiliation":[{"name":"Department of Botany, University of Buner, Swari 19290, Khyber Pakhtunkhwa, Pakistan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian","family":"Huang","sequence":"additional","affiliation":[{"name":"Key Laboratory of National Forestry and Grassland Administration on Silviculture in Loess Plateau, College of Forestry, Northwest A&F University, No. 3 Taicheng Road, Yangling 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Riaz, M., Raza, M.A., Saeed, A., Ahmed, M., and Hussain, T. (2021). Variations in Morphological Characters and Antioxidant Potential of Different Plant Parts of Four Ziziphus Mill. Species from the Cholistan. Plants, 10.","DOI":"10.3390\/plants10122734"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"262","DOI":"10.4103\/pm.pm_398_16","article-title":"Immunomodulatory, Cytotoxicity, and Antioxidant Activities of Roots of Ziziphus mauritiana","volume":"13","author":"Afzal","year":"2017","journal-title":"Pharmacogn. Mag."},{"key":"ref_3","first-page":"556158","article-title":"Ethno-Pharmacognosy and Diversity Encourage Conservation of Wild Ziziphus Species Collected from KP, Pakistan","volume":"21","author":"Ali","year":"2019","journal-title":"Agric. Res. Technol. Open Access J."},{"key":"ref_4","first-page":"1","article-title":"Phytochemistry of Ziziphus mauritiana; Its Nutritional and Pharmaceutical Potential","volume":"5","author":"Hussain","year":"2021","journal-title":"Sci. Inq. Rev."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"349","DOI":"10.21273\/HORTSCI16267-21","article-title":"Determination of Genome Size and Chromosome Number of a Ziziphus Species (Z. mauritiana Lam.) from Eastern Senegal","volume":"57","author":"Sakhanokho","year":"2022","journal-title":"HortScience"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"4597","DOI":"10.13057\/biodiv\/d240842","article-title":"Morphological Diversity of Indian Jujube (Ziziphus mauritiana) in Sumbawa Island, West Nusa Tenggara, Indonesia","volume":"24","author":"Nikmatullah","year":"2023","journal-title":"Biodiversitas J. Biol. Divers."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1016\/j.scienta.2019.01.041","article-title":"Exploitation of Diversity in Domesticated and Wild Ber (Ziziphus mauritiana Lam.) Germplasm for Conservation and Breeding in Pakistan","volume":"249","author":"Sharif","year":"2019","journal-title":"Sci. Hortic."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"100320","DOI":"10.1016\/j.fecs.2025.100320","article-title":"Design Strategy of Advanced Generation Breeding Population of Pinus tabuliformis Based on Genetic Variation and Inbreeding Level","volume":"13","author":"Zhou","year":"2025","journal-title":"For. Ecosyst."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1875","DOI":"10.1007\/s00425-019-03133-2","article-title":"Genetic Diversity of Ziziphus mauritiana Germplasm Based on SSR Markers and Ploidy Level Estimation","volume":"249","author":"Liang","year":"2019","journal-title":"Planta"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2701","DOI":"10.1093\/plphys\/kiae512","article-title":"Genomes of Autotetraploid Wild and Cultivated Ziziphus mauritiana Reveal Polyploid Evolution and Crop Domestication","volume":"196","author":"Guo","year":"2024","journal-title":"Plant Physiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2406","DOI":"10.1038\/s41598-018-20548-1","article-title":"Transcriptome Sequence Analysis and Mining of SSRs in Jhar Ber (Ziziphus Nummularia (Burm.f.) Wight & Arn) under Drought Stress","volume":"8","author":"Yadav","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"20767","DOI":"10.1038\/s41598-023-48022-7","article-title":"SSR Molecular Marker Developments and Genetic Diversity Analysis of Zanthoxylum nitidum (Roxb.) DC","volume":"13","author":"Zhu","year":"2023","journal-title":"Sci. Rep."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1939","DOI":"10.1007\/s12298-022-01249-6","article-title":"Genomic Constitution and Relationships of Ziziphus Species Collected from Malakand Division, Pakistan","volume":"28","author":"Uddin","year":"2022","journal-title":"Physiol. Mol. Biol. Plants"},{"key":"ref_14","first-page":"79","article-title":"Genetic Diversity in Jujube Germplasm (Ziziphus jujuba Mill.) Based on Morphological and Pomological Traits in Isfahan Province, Iran","volume":"6","author":"Tatari","year":"2016","journal-title":"Comun. Sci."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Jiang, F., Zhu, X., Wu, M., Chang, P., Wu, H., and Li, H. (2025). Domestication Has Reshaped Gene Families, Gene Expressions and Flavonoid Metabolites in Green Jujube (Ziziphus mauritiana Lam.) Fruit. Horticulturae, 11.","DOI":"10.3390\/horticulturae11080974"},{"key":"ref_16","unstructured":"Horwitz, W. (1960). Official Methods of Analysis of the Association of Official Agricultural Chemists, Association of Official Agricultural Chemists. [9th ed.]."},{"key":"ref_17","unstructured":"Ruck, J.A. (1963). Chemical Methods for Analysis of Fruit and Vegetable Products, Canada Department of Agriculture."},{"key":"ref_18","unstructured":"Mahajan, R.K., Gangopadhyay, K.K., Gunjeet, K., Dobhal, V.K., Srivastava, U., Gupta, P.N., and Pareek, S.K. (2002). Minimal Descriptors of Agri-Horticultural Crops, Part III: Fruit Crops, National Bureau of Plant Genetic Resources, Pusa Campus."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"50116","DOI":"10.19045\/bspab.2016.50116","article-title":"Fruit Morphological Attributes to Assess Genetic Diversity in Jujube (Ziziphus mauritiana L.) Germplasm of Bahawalpur","volume":"5","author":"Ahmad","year":"2016","journal-title":"Pure Appl. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"903","DOI":"10.1002\/fsn3.2721","article-title":"Selection of Superior Genotypes of Indian Jujube (Ziziphus mauritiana Lamk.) as Revealed by Fruit-related Traits","volume":"10","author":"Mirheidari","year":"2022","journal-title":"Food Sci. Nutr."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Meena, V.S., Singh, K., Shekhawat, N., Bhardwaj, R., Lal, H., Rani, K., Gupta, V., Kumar, A., Singh, A., and G\u00f3ra, J. (2023). Assessment of Genetic Variability for Fruit Nutritional Composition in the Ex-Situ Collection of Jujube (Ziziphus Spp.) Genotypes of Arid Regions of India. Horticulturae, 9.","DOI":"10.3390\/horticulturae9020210"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Abdel-Sattar, M., Al-Obeed, R.S., Rihan, H.Z., and El-Badan, G.E. (2024). Genetic Diversity and Relationships among Indian Jujube (Ziziphus mauritiana Lamk.) Cultivars Using Morphometric Characteristics, matK Barcoding, and ISSR Markers. Diversity, 16.","DOI":"10.3390\/d16060313"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1080\/15324982.2018.1424742","article-title":"Genetic Diversity of Ziziphus lotus Natural Populations from Algeria Based on Fruit Morphological Markers","volume":"32","author":"Boussaid","year":"2018","journal-title":"Arid Land Res. Manag."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"135","DOI":"10.9734\/ijpss\/2024\/v36i34408","article-title":"Correlation Studies in Ber (Ziziphus mauriatiana Lamk.) in Eastern Region of Uttar Pradesh, India","volume":"36","author":"Kumar","year":"2024","journal-title":"Int. J. Plant Soil Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"3424","DOI":"10.1016\/j.sjbs.2021.03.006","article-title":"Fruit Properties during the Harvest Period of Eleven Indian Jujube (Ziziphus mauritiana Lamk.) Cultivars","volume":"28","author":"Almutairi","year":"2021","journal-title":"Saudi J. Biol. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Bai, Y., Xie, J., Tong, T., Zhou, X., Yuan, Z., Zhang, Y., Li, X., and Wu, C. (2025). Genetic Diversity Analysis of Phenotypic Traits in Jujube Germplasm Resources. Agronomy, 15.","DOI":"10.3390\/agronomy15092063"},{"key":"ref_27","first-page":"97","article-title":"Biodiversity Evaluation among Wild Jujube (Ziziphus nummularia (Burm. F.) Population in Thal Desert of Pakistan","volume":"8","author":"Sharif","year":"2022","journal-title":"J. Innov. Sci."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Sun, M., Yaman, M., S\u00fcmb\u00fcl, A.T., and Demirel, F. (2024). Morphological, Biochemical Content, Antioxidant Activity and Molecular Characterization of Barberry Genotypes Growing Naturally in Central T\u00fcrkiye. Res. Sq.","DOI":"10.21203\/rs.3.rs-5301100\/v1"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Yahia, Y., Benabderrahim, M.A., Tlili, N., Bagues, M., and Nagaz, K. (2020). Bioactive Compounds, Antioxidant and Antimicrobial Activities of Extracts from Different Plant Parts of Two Ziziphus Mill. Species. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0232599"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1016\/j.scienta.2017.05.016","article-title":"Morphological Variability within and among Three Species of Ziziphus Genus Using Multivariate Analysis","volume":"222","author":"Norouzi","year":"2017","journal-title":"Sci. Hortic."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Khadivi, A. (2023). Morphological Characterization and Interspecific Variation among Five Species of Ziziphus Genus to Select Superiors in Iran. BMC Plant Biol., 23.","DOI":"10.1186\/s12870-023-04566-4"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Li, X., Zhang, L., Wei, X., Datta, T., Wei, F., and Xie, Z. (2024). Polyploidization: A Biological Force That Enhances Stress Resistance. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25041957"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Tossi, V.E., Mart\u00ednez Tosar, L.J., Laino, L.E., Iannicelli, J., Regalado, J.J., Escand\u00f3n, A.S., Baroli, I., Causin, H.F., and Pitta-\u00c1lvarez, S.I. (2022). Impact of Polyploidy on Plant Tolerance to Abiotic and Biotic Stresses. Front. Plant Sci., 13.","DOI":"10.3389\/fpls.2022.869423"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1093\/aob\/mcx079","article-title":"(Pat) Polyploidy and Interspecific Hybridization: Partners for Adaptation, Speciation and Evolution in Plants","volume":"120","author":"Alix","year":"2017","journal-title":"Ann. Bot."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1186\/s13578-021-00633-1","article-title":"Multiple Responses Contribute to the Enhanced Drought Tolerance of the Autotetraploid Ziziphus jujuba Mill. Var. Spinosa","volume":"11","author":"Meng","year":"2021","journal-title":"Cell Biosci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"108854","DOI":"10.1016\/j.scienta.2019.108854","article-title":"The \u201cPolyploid Effect\u201d in the Breeding of Aromatic and Medicinal Species","volume":"260","author":"Iannicelli","year":"2020","journal-title":"Sci. Hortic."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Li-hu, W., Wang, L., Ye, T., Zhao, J., Wang, L., Wei, H., Liu, P., and Liu, M. (2023). Autotetraploidization Alters Morphology, Photosynthesis, Cytological Characteristics and Fruit Quality in Sour Jujube (Ziziphus acidojujuba Cheng et Liu). Plants, 12.","DOI":"10.3390\/plants12051106"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"108679","DOI":"10.1016\/j.plaphy.2024.108679","article-title":"Integrated Anatomical Structure, Physiological, and Transcriptomic Analyses to Identify Differential Cold Tolerance Responses of Ziziphus jujuba Mill. \u2018Yueguang\u2019 and Its Autotetraploid \u2018Hongguang\u2019","volume":"211","author":"Zhu","year":"2024","journal-title":"Plant Physiol. Biochem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1186\/s43141-022-00458-6","article-title":"Assessment of Genetic Diversity and Population Structure in Wild Ziziphus Species from Northwest India Using SSR Marker Technique","volume":"21","author":"Sareen","year":"2023","journal-title":"J. Genet. Eng. Biotechnol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1007\/BF03321926","article-title":"Assessment of Genetic Diversity in Ziziphus mauritiana Using Inter-Simple Sequence Repeat Markers","volume":"16","author":"Singh","year":"2007","journal-title":"J. Plant Biochem. Biotechnol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1007\/s11295-016-1092-x","article-title":"Genetic Diversity of Indian Jujube Cultivars Using SCoT, ISSR, and rDNA Markers","volume":"13","author":"Singh","year":"2017","journal-title":"Tree Genet. Genomes"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"e14426","DOI":"10.1111\/ppl.14426","article-title":"Systematic Exploration of Ziziphus Species Collection and Gene Identification Influencing Physiological Traits Using Genotype-by-sequencing Technology","volume":"176","author":"Uddin","year":"2024","journal-title":"Physiol. Plant."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1002\/csc2.20080","article-title":"Assessment of Genetic Diversity in Ziziphus jujube Mill. Cultivars Derived from Northern China Using Inter-simple Sequence Repeat Markers","volume":"60","author":"Shen","year":"2020","journal-title":"Crop Sci."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"e05078","DOI":"10.1016\/j.heliyon.2020.e05078","article-title":"The Genetic Relationships of Indian Jujube (Ziziphus mauritiana Lam.) Cultivars Using SSR Markers","volume":"6","author":"Chiou","year":"2020","journal-title":"Heliyon"},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Huang, J., Yang, X., Zhang, C., Yin, X., Liu, S., and Li, X. (2015). Development of Chloroplast Microsatellite Markers and Analysis of Chloroplast Diversity in Chinese Jujube (Ziziphus jujuba Mill.) and Wild Jujube (Ziziphus acidojujuba Mill.). PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0134519"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1069","DOI":"10.1111\/jse.13065","article-title":"Comparative Analysis of 326 Chloroplast Genomes in Chinese Jujube (Ziziphus jujuba): Structural Variations, Horizontal Gene Transfer Events, and Evolutionary Patterns Impacting Its Domestication from Wild Jujube","volume":"62","author":"Yang","year":"2024","journal-title":"J. Syst. Evol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"31503","DOI":"10.1038\/srep31503","article-title":"Identifying the Genetic Diversity, Genetic Structure and a Core Collection of Ziziphus jujuba Mill. Var. Jujuba Accessions Using Microsatellite Markers","volume":"6","author":"Xu","year":"2016","journal-title":"Sci. Rep."}],"container-title":["Plants"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2223-7747\/15\/13\/1974\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,29]],"date-time":"2026-06-29T03:17:31Z","timestamp":1782703051000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2223-7747\/15\/13\/1974"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,26]]},"references-count":47,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["plants15131974"],"URL":"https:\/\/doi.org\/10.3390\/plants15131974","relation":{},"ISSN":["2223-7747"],"issn-type":[{"value":"2223-7747","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,26]]}}}