Phenotypic characteristics and analysis of Cypripedium in Heilongjiang Province

SHEN Caiquan, DONG Shang, LI Zhongyue, GUAN Yanhui, MU Liqiang

Journal of Nanjing Forestry University (Natural Sciences Edition） ›› 2026, Vol. 50 ›› Issue (2) : 85-95.

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南京林业大学学报（自然科学版）
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Journal of Nanjing Forestry University (Natural Sciences Edition） ›› 2026, Vol. 50 ›› Issue (2) : 85-95. DOI: 10.12302/j.issn.1000-2006.202410038

Phenotypic characteristics and analysis of Cypripedium in Heilongjiang Province

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Abstract

【Objective】This study aims to clarify the phylo genetic relationships and taxonomic boundaries of Cypripedium in Heilongjiang Province, where the genus exhibits extensive phenotypic variation, ambiguous species delimitation, and complex phylogenetic patterns.【Method】A total of 13 natural populations collected from the region were used as the research material, we measured 18 phenotypic traits across these 13 populations and performed phenotypic diversity and cluster analyses. Furthermore, we sequenced, assembled and annotated the nrITS region and three chloroplast DNA fragments (trnH—psbA, trnL—F, and rbcL). Nucleotide polymorphism was statistically analyzed, and phylogenetic trees were reconstructed using software packages DnaSP, PhyloSuite and FigTree, incorporating relevant sequences from the NCBI public database.【Result】Staminode characteristics (presence of mottling and size) serve as key traits for field identification of C. species in Heilongjiang Province. C. × ventricosum exhibits significant heterosis, with leaf size, plant height and stem diameter at ground level providing reliable morphological markers for its classification. Integrated phenotypic and phylogenetic analyses consistently demonstrated that: the white-and pink-flowered forms of C. macranthos constitute the same species; C. calceolus and C. shanxiense likely represent sister lineages derived from the same maternal ancestor; populations DB1, DB2 and DB5 of C. × ventricosum form a distinct clade potentially originating from C. calceolus as the maternal parent, while populations DB3, DB4, DB6, DB7 and DB8 cluster in another clade likely descended from C. macranthos, with population DB6 showing evidence of possible backcrossing with C. macranthos; C. guttatum remains phylogenetically distant from all other studied species.【Conclusion】Inferring genetic relationships in Cypripedium, particularly for hybrid taxa, cannot rely solely on a single phenotypic trait or single DNA marker. A comprehensive approach integrating phenotypic data with both chloroplast and nuclear DNA sequences is essential for accurately elucidating phylogenetic relationships and taxonomic boundaries.

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Cypripedium plants / phenotypic characteristics / phylogenetic relationship / plant taxonomy / chloroplast genes

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SHEN Caiquan , DONG Shang , LI Zhongyue , et al . Phenotypic characteristics and analysis of Cypripedium in Heilongjiang Province[J]. Journal of Nanjing Forestry University (Natural Sciences Edition）. 2026, 50(2): 85-95 https://doi.org/10.12302/j.issn.1000-2006.202410038

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