目的 建立不同生長年限實生芍藥Paeonia lactiflora指紋圖譜，同時測定6種差異標(biāo)志物成分含量及15種農(nóng)藝性狀，篩選實生芍藥與生長年限關(guān)聯(lián)的關(guān)鍵化學(xué)成分和主要農(nóng)藝性狀特征屬性，以了解不同生長年限實生芍藥的生長發(fā)育規(guī)律。方法 5組實生芍藥材料，每組隨機(jī)取樣10株，共建立50株不同生長年限的實生芍藥HPLC指紋圖譜，并測定沒食子酸、氧化芍藥苷、芍藥內(nèi)酯苷、芍藥苷、五沒食子酰葡萄糖、苯甲酰芍藥苷6種差異標(biāo)志物含量；同時測定15種農(nóng)藝性狀（株高、冠幅、分枝數(shù)、葉片數(shù)、葉面積、地上鮮質(zhì)量、開花數(shù)、主根長、主根粗、主根數(shù)、須根長、須根數(shù)、地下鮮質(zhì)量、主根鮮質(zhì)量、須根鮮質(zhì)量）。利用層次聚類分析（hierarchical cluster analysis，HCA）、主成分分析（principal component analysis，PCA）、正交偏最小二乘法-判別分析（orthogonal partial least squares-discriminant analysis，OPLS-DA）和Spearman相關(guān)性分析，篩選與生長年限關(guān)聯(lián)的關(guān)鍵化學(xué)成分和主要農(nóng)藝性狀。結(jié)果 50株實生芍藥共有27個共有峰，其中S0、S1、S2、S3、S4分別標(biāo)定出10、10、13、16、19個特征峰；通過PCA和OPLS-DA分析，篩選出7個差異性化學(xué)成分標(biāo)志物，其變量重要性投影（variable importance in projection，VIP）值依次為峰12（芍藥苷）＞峰10（芍藥內(nèi)酯苷）＞峰24＞峰3（沒食子酸）＞峰23＞峰19（五沒食子酰葡萄糖）＞峰22（VIP＞1）。含量測定結(jié)果表明，芍藥苷、氧化芍藥苷、五沒食子酰葡萄糖含量隨生長年限呈顯著增加趨勢；芍藥內(nèi)酯苷含量先降低后升高，苯甲酰芍藥苷緩慢升高但差異不顯著，趨于穩(wěn)定；沒食子酸含量在第1年顯著增加，之后均趨于穩(wěn)定；其中VIP值為芍藥苷＞芍藥內(nèi)酯苷（VIP＞1）。實生芍藥的農(nóng)藝性狀分析表明，PCA和OPLS-DA篩選出3個差異標(biāo)志性農(nóng)藝性狀，其VIP值依次為須根長＞開花數(shù)＞主根粗（VIP＞1）。結(jié)論 隨著生長年限的增加，實生芍藥根部藥材的共有峰數(shù)量增多，相似度提高，化學(xué)組成的穩(wěn)定性和一致性增強(qiáng)；農(nóng)藝性狀方面，地上部分體積增大，地下部分根系生長、分枝、擴(kuò)展，芍藥總生物量快速積累。芍藥苷可作為區(qū)分不同生長年限實生芍藥的關(guān)鍵差異性化學(xué)成分標(biāo)志物；須根長、開花數(shù)、主根粗可作為區(qū)分不同生長年限實生芍藥的重要農(nóng)藝性狀標(biāo)志物。

Objective To establish fingerprint profiles of Paeonia lactiflora in different growth ages and measure six differential marker compounds content and 15 agronomic traits. To screen the key chemical components and main agronomic characteristics of P. lactiflora related to the growth ages in order to understand the growth and development law of P. lactiflora in different growth ages. Methods For five groups of P. lactiflora (10 plants per group), HPLC fingerprint profiles were created for 50 plants of different growth durations. The content of six differential marker compounds (gallic acid, oxypaeoniflorin, albiflorin, paeoniflorin, pentagalloylglucose and benzoylpaeoniflorin) and 15 agronomic traits (plant height, crown width, number of branches, number of leaves, leaf area, above ground fresh quality, number of flowers, length of taproot, thickness of taproot, number of taproot, length of fibril, number of fibril, underground fresh quality, fresh quality of taproot, fresh quality of fibril) were measured. Hierarchical cluster analysis (HCA), principal component analysis (PCA), orthogonal partial least squares-discriminant analysis (OPLS-DA) and Spearman correlation analysis were used to identify key chemical components and major agronomic traits associated with growth duration. Results Among the 50 P. lactiflora, 27 common peaks were identified. S0, S1, S2, S3 and S4 had 10, 10, 13, 16 and 19 characteristic peaks, respectively. PCA and OPLS-DA identified seven differential chemical markers, and their variable importance in projection (VIP) values were peak 12 (paeoniflorin) > peak 10 (albiflorin) > peak 24 > peak 3 (gallic acid) > peak 23 > peak 19 (pentagalloylglucose) > peak 22 (VIP > 1). The results of content determination showed that the content of paeoniflorin, oxypaeoniflorin and pentagalloylglucose significantly increased with growth duration. Albiflorin decreased initially then increased, benzoylpaeoniflorin increased slowly but not significantly and tended to be stable, and gallic acid increased significantly in the first year then stabilized. The VIP value was paeoniflorin > albiflorin (VIP > 1). The analysis of agronomic traits of P. lactiflora showed that PCA and OPLS-DA selected three differential signature agronomic traits, and their VIP values were length of fibril > number of flowers > thickness of taproot (VIP > 1). Conclusion With increased growth duration, P. lactiflora show more common peaks, increased similarity, and increased stability and consistency of chemical composition. . In terms of agronomic traits, the volume of the above-ground part increases, the roots of the underground part grow, branch and expand, and the total biomass accumulated rapidly. Paeoniflorin can be used as a key differential chemical marker to distinguish P. lactiflora in different growth durations, while length of fibril, number of flowers, thickness of taproot are important agronomic markers to distinguish P. lactiflora in different growth durations.

R286.2

四川省“十四五”農(nóng)作物及畜禽育種攻關(guān)項目（2021YFYZ0012）；四川省科技計劃資助（2023YFQ0112）；四川省財政自主創(chuàng)新專項項目（2022ZZCX077）；國家現(xiàn)代農(nóng)業(yè)產(chǎn)業(yè)技術(shù)體系建設(shè)專項項目（CARS-21）