{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,1]],"date-time":"2026-02-01T03:12:34Z","timestamp":1769915554980,"version":"3.49.0"},"reference-count":46,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,1,29]],"date-time":"2025-01-29T00:00:00Z","timestamp":1738108800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Plant Sci."],"abstract":"<jats:p>Alfalfa is one of the primary forages, and its yield is largely dependent on the development of its leaf. In this study, to elucidate the mechanism of compound leaf development, we isolated and examined the alfalfa phenotype Chuancao No.7, exhibiting five leaflets. The agronomic traits of the Chuancao No.7 including the leaf blade area, leaf\/stem ratio, total fresh weight, and dry weight showed significant increases compared to those of the wild-type. Analyses of forage quality traits indicated significant differences in crude protein (CP), acid detergent fiber (ADF), crude fat (CF), water-soluble sugars (WSS), carbon content, ash composition content, and phosphorus content between Chuancao No.7 alfalfa and wild-type. Transcriptomic profile analysis revealed that differentially expressed genes were identified in the cytokinin (CK) signaling pathway. Both exogenous treatment and endogenous CK content detection indicated that cytokinin played a key role in the development of the alfalfa compound leaf. These results serve as a valuable resource for optimizing the forage quality and exploring the excellent germplasm of alfalfa.<\/jats:p>","DOI":"10.3389\/fpls.2025.1460205","type":"journal-article","created":{"date-parts":[[2025,1,29]],"date-time":"2025-01-29T06:58:51Z","timestamp":1738133931000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":4,"title":["Transcriptomic and physiological analyses reveal that cytokinin is involved in the compound leaf development of alfalfa"],"prefix":"10.3389","volume":"16","author":[{"given":"Hongyao","family":"Mei","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiajun","family":"Yan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xuexin","family":"Jia","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Weilin","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shuangshuang","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ruiqi","family":"Sun","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hongjiao","family":"Jiang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lijun","family":"Xie","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chuanen","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shiqie","family":"Bai","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lu","family":"Han","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2025,1,29]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"61","DOI":"10.1016\/j.pbi.2014.10.007","article-title":"Compound leaf development in model plant species","volume":"23","author":"Bar","year":"2015","journal-title":"Curr. Opin. Plant Biol."},{"key":"B2","doi-asserted-by":"publisher","first-page":"1752","DOI":"10.3390\/plants11131752","article-title":"Optimization of alfalfa-based mixed cropping with winter wheat and ryegrass in terms of forage yield and quality traits","volume":"11","author":"Bo","year":"2022","journal-title":"Plants"},{"key":"B3","doi-asserted-by":"publisher","first-page":"3369","DOI":"10.1105\/tpc.107.052886","article-title":"Compound leaf development and evolution in the legumes","volume":"19","author":"Champagne","year":"2007","journal-title":"Plant Cell."},{"key":"B4","doi-asserted-by":"publisher","first-page":"145","DOI":"10.1038\/s41586-022-05529-9","article-title":"NLR surveillance of pathogen interference with hormone receptors induces immunity","volume":"613","author":"Chen","year":"2022","journal-title":"Nature"},{"key":"B5","doi-asserted-by":"publisher","first-page":"998","DOI":"10.1111\/pce.13494","article-title":"Cytokinin action in response to abiotic and biotic stresses in plants","volume":"42","author":"Cortleven","year":"2019","journal-title":"Plant Cell Environ."},{"key":"B6","doi-asserted-by":"publisher","first-page":"2090","DOI":"10.1104\/pp.100.4.2090","article-title":"Inhibition of growth of cultured tobacco cells at low concentrations of lovastatin is reversed by cytokinin","volume":"100","author":"Crowell","year":"1992","journal-title":"Plant Physiol."},{"key":"B7","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1007\/s11103-008-9307-7","article-title":"Characterization of KNOX genes in Medicago truncatula","volume":"67","author":"Di Giacomo","year":"2008","journal-title":"Plant Mol. Biol."},{"key":"B8","doi-asserted-by":"publisher","first-page":"350","DOI":"10.1021\/ac60111a017","article-title":"Colorimetric method for determination of sugars and related substances","volume":"28","author":"DuBois","year":"1956","journal-title":"Analytical Chem."},{"key":"B9","doi-asserted-by":"publisher","first-page":"1750","DOI":"10.3390\/s23041750","article-title":"Practical considerations for using the neospectra-scanner handheld near-infrared reflectance spectrometer to predict the nutritive value of undried ensiled forage","volume":"23","author":"Feng","year":"2023","journal-title":"Sensors (Basel)."},{"key":"B10","doi-asserted-by":"publisher","first-page":"1134","DOI":"10.4161\/psb.5.9.12640","article-title":"Palmate-like pentafoliata1 encodes a novel Cys(2)His(2) zinc finger transcription factor essential for compound leaf morphogenesis in Medicago truncatula","volume":"5","author":"Ge","year":"2010","journal-title":"Plant Signaling Behavior."},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.3389\/fpls.2022.1047294","article-title":"Forage quality and physiological performance of mowed alfalfa (Medicago sativa L.) subjected to combined light quality and drought","volume":"13","author":"He","year":"2022","journal-title":"Front. Plant Science."},{"key":"B12","doi-asserted-by":"publisher","first-page":"882","DOI":"10.1002\/jsfa.7810","article-title":"Extraction, composition, and functional properties of dried alfalfa (Medicago sativa L.) leaf protein","volume":"97","author":"Hojilla-Evangelista","year":"2017","journal-title":"J. Sci. Food Agric."},{"key":"B13","doi-asserted-by":"publisher","first-page":"2735","DOI":"10.3390\/plants11202735","article-title":"Characterization of forage quality, phenolic profiles, and antioxidant activity in alfalfa (Medicago sativa L.)","volume":"11","author":"Horvat","year":"2022","journal-title":"Plants"},{"key":"B14","doi-asserted-by":"publisher","first-page":"414","DOI":"10.1007\/s00203-022-03046-x","article-title":"Effects of urea supplementation on the nutritional quality and microbial community of alfalfa (Medicago sativa L.) silage","volume":"204","author":"Hou","year":"2022","journal-title":"Arch. Microbiol."},{"key":"B15","doi-asserted-by":"publisher","first-page":"1560","DOI":"10.1016\/j.cub.2005.07.023","article-title":"KNOX action in Arabidopsis is mediated by coordinate regulation of cytokinin and gibberellin activities","volume":"15","author":"Jasinski","year":"2005","journal-title":"Curr. Biol."},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1002\/cft2.20076","article-title":"Forage nutritive value of modern alfalfa cultivars","volume":"6","author":"Jungers","year":"2020","journal-title":"Crop Forage Turfgrass Management."},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.3389\/fpls.2014.00362","article-title":"Leaf development: a cellular perspective","volume":"5","author":"Kalve","year":"2014","journal-title":"Front. Plant Science."},{"key":"B18","doi-asserted-by":"publisher","first-page":"dev149344","DOI":"10.1242\/dev.149344","article-title":"Cytokinin signaling in plant development","volume":"145","author":"Kieber","year":"2018","journal-title":"Development"},{"key":"B19","doi-asserted-by":"publisher","first-page":"2193","DOI":"10.1093\/jxb\/erf077","article-title":"A trans-zeatin riboside in root xylem sap negatively regulates adventitious root formation on cucumber hypocotyls","volume":"53","author":"Kuroha","year":"2002","journal-title":"J. Exp. Botany."},{"key":"B20","doi-asserted-by":"publisher","first-page":"1859","DOI":"10.1105\/tpc.6.12.1859","article-title":"A knotted1-like homeobox gene in Arabidopsis is expressed in the vegetative meristem and dramatically alters leaf morphology when overexpressed in transgenic plants","volume":"6","author":"Lincoln","year":"1994","journal-title":"Plant Cell."},{"key":"B21","doi-asserted-by":"publisher","first-page":"1111","DOI":"10.1016\/j.plantsci.2006.01.015","article-title":"Cytokinin-induced changes in CLAVATA1 and WUSCHEL expression temporally coincide with altered floral development in Arabidopsis","volume":"170","author":"Lindsay","year":"2006","journal-title":"Plant Science."},{"key":"B22","doi-asserted-by":"publisher","first-page":"2988","DOI":"10.1038\/s41467-024-47362-w","article-title":"Rewiring of a KNOXI regulatory network mediated by UFO underlies the compound leaf development in Medicago truncatula","volume":"15","author":"Lu","year":"2024","journal-title":"Nat. Commun."},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.7717\/peerj.10874","article-title":"Phenotype study of multifoliolate leaf formation inTrifolium alexandrinumL","volume":"9","author":"Malaviya","year":"2021","journal-title":"PeerJ"},{"key":"B24","doi-asserted-by":"publisher","first-page":"3007","DOI":"10.1105\/tpc.105.035451","article-title":"Multiple type-B response regulators mediate cytokinin signal transduction in Arabidopsis","volume":"17","author":"Mason","year":"2005","journal-title":"Plant Cell."},{"key":"B25","doi-asserted-by":"publisher","first-page":"1365","DOI":"10.1105\/tpc.021477","article-title":"Histidine kinase homologs that act as cytokinin receptors possess overlapping functions in the regulation of shoot and root growth in Arabidopsis","volume":"16","author":"Nishimura","year":"2004","journal-title":"Plant Cell."},{"key":"B26","doi-asserted-by":"publisher","first-page":"14114","DOI":"10.3390\/ijms232214114","article-title":"Arabidopsis thaliana SHOOT MERISTEMLESS substitutes for medicago truncatula SINGLE LEAFLET1 to form complex leaves and petals","volume":"23","author":"Pautot","year":"2022","journal-title":"Int. J. Mol. Sci."},{"key":"B27","doi-asserted-by":"publisher","first-page":"3929","DOI":"10.1105\/tpc.111.089128","article-title":"Regulation of compound leaf development in Medicago truncatula by fused compound leaf1, a class M KNOX gene","volume":"23","author":"Peng","year":"2011","journal-title":"Plant Cell."},{"key":"B28","doi-asserted-by":"publisher","first-page":"1161","DOI":"10.1071\/FP21031","article-title":"Phosphorus and selenium uptake, root morphology, and carboxylates in the rhizosheath of alfalfa (Medicago sativa) as affected by localised phosphate and selenite supply in a split-root system","volume":"48","author":"Peng","year":"2021","journal-title":"Funct. Plant Biol."},{"key":"B29","doi-asserted-by":"publisher","first-page":"40","DOI":"10.1105\/tpc.105.037796","article-title":"Arabidopsis cytokinin receptor mutants reveal functions in shoot growth, leaf senescence, seed size, germination, root development, and cytokinin metabolism","volume":"18","author":"Riefler","year":"2006","journal-title":"Plant Cell."},{"key":"B30","doi-asserted-by":"publisher","first-page":"747","DOI":"10.2527\/jas1978.473747x","article-title":"Proposed hay grading standards based on laboratory analyses for evaluating quality","volume":"47","author":"Rohweder","year":"1978","journal-title":"J. Anim. Science."},{"key":"B31","doi-asserted-by":"publisher","first-page":"557","DOI":"10.1046\/j.1365-313X.1999.00472.x","article-title":"Increased steady state mRNA levels of the STM and KNAT1 homeobox genes in cytokinin overproducing Arabidopsis thaliana indicate a role for cytokinins in the shoot apical meristem","volume":"18","author":"Rupp","year":"2002","journal-title":"Plant J."},{"key":"B32","doi-asserted-by":"publisher","first-page":"431","DOI":"10.1146\/annurev.arplant.57.032905.105231","article-title":"CYTOKININS: activity, biosynthesis, and translocation","volume":"57","author":"Sakakibara","year":"2006","journal-title":"Annu. Rev. Plant Biol."},{"key":"B33","doi-asserted-by":"publisher","first-page":"44","DOI":"10.1105\/tpc.114.133595","article-title":"The yin-yang of hormones: cytokinin and auxin interactions in plant development","volume":"27","author":"Schaller","year":"2015","journal-title":"Plant Cell."},{"key":"B34","doi-asserted-by":"publisher","first-page":"2125","DOI":"10.1105\/tpc.111.085340","article-title":"STENOFOLIA regulates blade outgrowth and leaf vascular patterning in Medicago truncatula and Nicotiana sylvestris","volume":"23","author":"Tadege","year":"2011","journal-title":"Plant Cell."},{"key":"B35","doi-asserted-by":"publisher","first-page":"4368","DOI":"10.1105\/tpc.111.090050","article-title":"Positive autoregulation of a KNOX gene is essential for shoot apical meristem maintenance in rice","volume":"23","author":"Tsuda","year":"2011","journal-title":"Plant Cell."},{"key":"B36","doi-asserted-by":"publisher","first-page":"79","DOI":"10.17557\/tjfc.43607","article-title":"Agro-morphological and forage quality traits of selected alfalfa populations and their application in breeding","volume":"19","author":"Tucak","year":"2014","journal-title":"Turkish J. Field Crops."},{"key":"B37","doi-asserted-by":"publisher","first-page":"241","DOI":"10.1038\/350241a0","article-title":"The developmental gene knotted-1 is a member of a maize homeobox gene family","volume":"350","author":"Vollbrecht","year":"1991","journal-title":"Nature"},{"key":"B38","doi-asserted-by":"publisher","first-page":"1759","DOI":"10.1104\/pp.108.117044","article-title":"Control of compound leaf development by floricaula\/leafyorthologsingle leaflet1inmedicago truncatula","volume":"146","author":"Wang","year":"2008","journal-title":"Plant Physiol."},{"key":"B39","doi-asserted-by":"publisher","first-page":"419","DOI":"10.1111\/jipb.12034","article-title":"Multiple components are integrated to determine leaf complexity in lotus japonicus","volume":"55","author":"Wang","year":"2013","journal-title":"J. Integr. Plant Biol."},{"key":"B40","doi-asserted-by":"publisher","first-page":"48","DOI":"10.3390\/agronomy14010048","article-title":"Alfalfa (Medicago sativa L.) nitrogen utilization, yield and quality respond to nitrogen application level with center pivot fertigation system","volume":"14","author":"Wang","year":"2024","journal-title":"Agronomy-Basel"},{"key":"B41","doi-asserted-by":"publisher","first-page":"10487","DOI":"10.1073\/pnas.171304098","article-title":"Regulation of plant growth by cytokinin","volume":"98","author":"Werner","year":"2001","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"B42","doi-asserted-by":"publisher","first-page":"118","DOI":"10.1038\/s41438-021-00558-3","article-title":"The diverse roles of cytokinins in regulating leaf development","volume":"8","author":"Wu","year":"2021","journal-title":"Hortic. Res."},{"key":"B43","doi-asserted-by":"publisher","first-page":"1967","DOI":"10.3390\/genes12121967","article-title":"Combined transcriptome and metabolome analysis of alfalfa response to thrips infection","volume":"12","author":"Zhang","year":"2021","journal-title":"Genes (Basel)."},{"key":"B44","doi-asserted-by":"publisher","first-page":"1464","DOI":"10.1105\/tpc.114.123885","article-title":"STM\/BP-Like KNOXI is uncoupled from ARP in the regulation of compound leaf development in Medicago truncatula","volume":"26","author":"Zhou","year":"2014","journal-title":"Plant Cell."},{"key":"B45","doi-asserted-by":"publisher","first-page":"1483","DOI":"10.1104\/pp.111.185140","article-title":"From model to crop: functional analysis of a STAY-GREEN gene in the model legume Medicago truncatula and effective use of the gene for alfalfa improvement","volume":"157","author":"Zhou","year":"2011","journal-title":"Plant Physiol."},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.4161\/psb.24737","article-title":"Cytokinin inhibition of leaf senescence","volume":"8","author":"Zwack","year":"2014","journal-title":"Plant Signaling Behavior."}],"container-title":["Frontiers in Plant Science"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fpls.2025.1460205\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,1,29]],"date-time":"2025-01-29T06:58:58Z","timestamp":1738133938000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fpls.2025.1460205\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,29]]},"references-count":46,"alternative-id":["10.3389\/fpls.2025.1460205"],"URL":"https:\/\/doi.org\/10.3389\/fpls.2025.1460205","relation":{},"ISSN":["1664-462X"],"issn-type":[{"value":"1664-462X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,1,29]]},"article-number":"1460205"}}