目的 基于簡化基因組，建立一種準(zhǔn)確鑒定藥用芍藥Paeonia lactiflora栽培品種的單核苷酸多態(tài)性（single nucleotide polymorphism，SNP）分子標(biāo)記方法。方法 利用特異位點擴增片段測序（specific-locus smplified fragment sequencing，SLAF-seq）技術(shù)對浙江、安徽、四川和湖南的13個藥用芍藥栽培品種進(jìn)行SNP標(biāo)記開發(fā)。通過多個指標(biāo)如雜合率、缺失率和多態(tài)性來篩選候選標(biāo)記，并通過遺傳多樣性、群體結(jié)構(gòu)和主成分分析等方法進(jìn)一步評估候選標(biāo)記。最后采用能夠區(qū)分藥用芍藥栽培品種的最簡SNP組合繪制指紋圖譜。結(jié)果 通過SLAF-seq測序共獲得3 703 329個SLAF標(biāo)簽，其中多態(tài)性標(biāo)簽有60 749個，共獲得412 873個群體SNP標(biāo)記，最后篩選的47個核心SNP標(biāo)記位點表現(xiàn)出高多態(tài)性，可以很好地區(qū)分藥用芍藥栽培品種。結(jié)論 基于簡化基因組，建立了主要藥用芍藥栽培品種的SNP標(biāo)記開發(fā)和指紋圖譜繪制的方法，為藥用芍藥的品種鑒定和白芍藥材的臨床應(yīng)用提供借鑒。

Objective Based on reduced-representation genome, we developed a single nucleotide polymorphism (SNP) molecular marker method for the accurate identification of medicinal cultivars of Paeonia lactiflora. Methods SNP markers were developed for 13 medicinal cultivars of P. lactiflora from Zhejiang, Anhui, Sichuan and Hunan provinces based on specific-locus smplified fragment sequencing (SLAF-seq) technology. Candidate markers were screened by multiple indicators such as heterozygosity rate, deletion rate and polymorphism, and further evaluated by genetic diversity, population structure and principal component analysis. Finally, fingerprinting was performed using the simplest SNP combination that could distinguish the medicinal cultivars of P. lactiflora. ResultsA total of 3 703 329 SLAF labels were obtained by SLAF-seq sequencing, including 60 749 polymorphic labels, and 412 873 population SNP markers were obtained. Finally, a total of 47 core SNP marker sites selected showed high polymorphism, and the SNP fingerprint drawn by this method could accurately distinguish medicinal cultivars of P. lactiflora. Conclusion A method of SNP marker development and fingerprint mapping for medicinal cultivars of P. lactiflora based on reduced-representation genome was established, which could provide reference for variety identification of P. lactiflora and clinical application of Cynanchum otophyllum.

R286.12

國家重點研發(fā)計劃項目(2022YFC3500903);中國中醫(yī)科學(xué)院科技創(chuàng)新工程項目(CI2024E001);安徽省高校自然科學(xué)研究重點項目(2023AH050854)