An efficient method for tissue-specific protoplast isolation suitable for single-cell RNA sequencing and transient gene expression analysis in saffron (Crocus sativus L.).

Xiaoyuan Xi, Jia Song, Mengqing Feng, Xiaodong Qian, Heng Sun, Liqin Li

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Article in BMC biotechnology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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6 authors.

Xiaoyuan XiTCM (Traditional Chinese Medicine) Key Laboratory Cultivation Base of Zhejiang Province for the Development and Clinical Transformation of Immunomodulatory drugs, Huzhou Central Hospital, Fifth School of Clinical Medicine of Zhejiang, Chinese Medical University, 1558 Sanhuan North Road, Huzhou, Zhejiang, 313000, China.

Jia SongTCM (Traditional Chinese Medicine) Key Laboratory Cultivation Base of Zhejiang Province for the Development and Clinical Transformation of Immunomodulatory drugs, Huzhou Central Hospital, Fifth School of Clinical Medicine of Zhejiang, Chinese Medical University, 1558 Sanhuan North Road, Huzhou, Zhejiang, 313000, China.

Mengqing FengSchool of Pharmacy, Hangzhou Normal University, Hangzhou, Zhejiang, China.

Xiaodong QianTCM (Traditional Chinese Medicine) Key Laboratory Cultivation Base of Zhejiang Province for the Development and Clinical Transformation of Immunomodulatory drugs, Huzhou Central Hospital, Fifth School of Clinical Medicine of Zhejiang, Chinese Medical University, 1558 Sanhuan North Road, Huzhou, Zhejiang, 313000, China.

Heng SunTCM (Traditional Chinese Medicine) Key Laboratory Cultivation Base of Zhejiang Province for the Development and Clinical Transformation of Immunomodulatory drugs, Huzhou Central Hospital, Fifth School of Clinical Medicine of Zhejiang, Chinese Medical University, 1558 Sanhuan North Road, Huzhou, Zhejiang, 313000, China.

Liqin LiTCM (Traditional Chinese Medicine) Key Laboratory Cultivation Base of Zhejiang Province for the Development and Clinical Transformation of Immunomodulatory drugs, Huzhou Central Hospital, Fifth School of Clinical Medicine of Zhejiang, Chinese Medical University, 1558 Sanhuan North Road, Huzhou, Zhejiang, 313000, China. liliqin@hzhospital.com.

Funding

National Natural Science Foundation of China 32570418National Natural Science Foundation of China 82304668Zhejiang Provincial Traditional Chinese Medicine Science and Technology Program 2026ZF85

6 · The paper itself

Abstract

backgroundProtoplasts have been widely utilized in tissue culture, single-cell RNA sequencing, and transient gene expression analyses. However, the primary challenge in their preparation lies in the efficient removal of rigid cell walls to obtain high-quality plant cells. The aim of this study is to establish an effective method for protoplast isolation from saffron (Crocus sativus L.), thereby accelerating advances in molecular biology research in this species.

resultTissue-specific protoplasts were successfully isolated from apical buds, petals, leaves, and roots of saffron, with yields of approximately 8.81 × 10

conclusionThis study presents the first detailed report on protoplast isolation, evaluation, and application in saffron. By overcoming the technical challenges of tissue-specific protoplast isolation from diverse organs, we established a highly efficient method that enables both the development of novel germplasm and mechanistic insights into saffron organogenesis, growth, and development.

Indexed as

CrocusProtoplastsSequence Analysis, RNASingle-Cell AnalysisSingle-Cell Gene Expression AnalysisEfficientSaffronSingle-cell RNA sequencingTissue-specific protoplast isolationTransient gene expression

Identifiers

PMID42421006

DOI10.1186/s12896-026-01187-1

PMCPMC13352814

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