Article(id=1321826884086780420, tenantId=1146029695717560320, journalId=1303687221787852879, issueId=1321826840323412132, articleNumber=null, orderNo=null, doi=10.7606/j.issn.1009-1041.2026.10.08, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1769702400000, receivedDateStr=2026-01-30, revisedDate=1772640000000, revisedDateStr=2026-03-05, acceptedDate=null, acceptedDateStr=null, onlineDate=1793077687522, onlineDateStr=2026-10-27, pubDate=1791993600000, pubDateStr=2026-10-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1793077687522, onlineIssueDateStr=2026-10-27, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1793077687522, creator=13701087609, updateTime=1793077687522, updator=13701087609, issue=Issue{id=1321826840323412132, tenantId=1146029695717560320, journalId=1303687221787852879, year='2026', volume='46', issue='10', pageStart='F2', pageEnd='1412', issueExtLink='null', onlineDate='null', pubDate='1791993600000', pubDateStr='2026-10-15', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=null, createTime=1793077677088, creator='13701087609', updateTime=1793077871239, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1321827654731412124, tenantId=1146029695717560320, journalId=1303687221787852879, issueId=1321826840323412132, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1321827654731412125, tenantId=1146029695717560320, journalId=1303687221787852879, issueId=1321826840323412132, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=1338, endPage=1345, ext={EN=ArticleExt(id=1321826884304884229, articleId=1321826884086780420, tenantId=1146029695717560320, journalId=1303687221787852879, language=EN, title=Optimization and Establishment of a Protocol for Protoplast Isolation from Barley Leaves, columnId=1305133119129023428, journalTitle=Journal of Triticeae Crops, columnName=Genetics & Breeding, runingTitle=null, highlight=null, articleAbstract=

To establish an efficient protocol for isolating high-quality protoplasts from barley leaf epidermal tissue, thereby providing technical support for studies on stomatal development, gene functional validation, and molecular breeding for stress resistance, this study used leaves of barley (Hordeum vulgare ‘Golden Promise’) at the three-leaf stage as experimental material. An orthogonal experimental design was employed to systematically optimize key parameters, including the concentrations of Cellulase R-10 and Macerozyme R-10, digestion time, and osmotic pressure. The results showed that enzyme concentration and digestion time significantly affected protoplast yield and morphology. The low-concentration enzyme combination (1.5% Cellulase R-10+0.75% Macerozyme R-10) failed to effectively release protoplasts. Increased enzyme concentrations (2% Cellulase R-10+1% Macerozyme R-10) improved yield, but levels remained relatively low. When the Macerozyme R-10 concentration was increased to 1.5% combined with a 6 h digestion, cell wall degradation was most complete, leading to significantly enhanced yield and improved morphological integrity. Osmotic pressure optimization revealed that increasing the mannitol concentration from 54.7 g·kg^-1 to 98.4 g·kg^-1 (resulting in an enzyme solution osmolality slightly higher than the leaf tissue osmotic potential of 630 mmol·kg^-1) increased protoplast yield from 4.8×10⁵ cells·mL^-1 to 5.8×10⁵ cells·mL^-1. Furthermore, protoplast morphology transitioned from irregular shapes with debris to intact, uniformly spherical forms. The optimal conditions were determined to be 2% Cellulase R-10, 1.5% Macerozyme R-10, a 6 h digestion time, and a mannitol concentration of 98.4 g·kg^-1, under which the protoplast yield was highest and the protoplasts were intact and uniformly spherical. This system is not only applicable to the lower epidermis of barley leaves but is also highly efficient for mesophyll tissue, providing reliable technical support for related cell biology and molecular biology research.

, authors=Xiaoyu Zhang, Jingjing Chen, Yiling Peng, Fenglin Deng, Yuan Qin, Fanrong Zeng, authorsList=Xiaoyu Zhang, Jingjing Chen, Yiling Peng, Fenglin Deng, Yuan Qin, Fanrong Zeng, authorCompany=null, correspAuthors=Yuan Qin, Fanrong Zeng, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1321826884732703238, articleId=1321826884086780420, tenantId=1146029695717560320, journalId=1303687221787852879, language=CN, title=大麦叶片原生质体制备条件的优化与体系建立, columnId=1305133119284212678, journalTitle=麦类作物学报, columnName=遗传育种, runingTitle=null, highlight=null, articleAbstract=

为建立一套稳定、高效的大麦下表皮原生质体制备体系，以啤酒大麦Golden Promise三叶一心期叶片为材料，采用正交实验设计，系统优化了原生质制备过程中的纤维素酶R-10与离析酶R-10浓度、酶解时间、渗透压等关键参数。结果表明，酶浓度与酶解时间显著影响原生质体产量与形态。低浓度酶组合（1.5%纤维素酶+0.75%离析酶）无法有效释放原生质体；提高酶浓度（2%纤维素酶+1%离析酶）后原生质体产量有所提升，但仍较低；当离析酶浓度增至1.5%并结合6 h酶解时，细胞壁降解最充分，原生质体产量与完整性显著提高。渗透压优化后，当甘露醇浓度从54.7 g·kg^-1提高至98.4 g·kg^-1（酶解液渗透压略高于组织渗透势630 mmol·kg^-1）时，原生质体产量从4.8×10⁵ cells·mL^-1提升至5.8×10⁵ cells·mL^-1，且形态由不规则伴碎片转变为完整规则的圆球形。最终确定最优组合为2%纤维素酶R-10、1.5%离析酶R-10、酶解6 h、98.4 g·kg^-1甘露醇，此条件下原生质体产量最高且形态完整，呈规则圆球形。该体系不仅适用于大麦叶片下表皮组织，对叶肉组织同样高效，可为大麦相关细胞生物学与分子生物学研究提供可靠的技术支持。

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