基于叶色表型和花青苷积累特征的牡丹红色叶性状解析

doi:10.3864/j.issn.0578-1752.2025.23.018

摘要/Abstract

摘要：

【目的】 牡丹不仅具有良好的观花价值，其新生叶片常常呈现紫红色，是园林中的春色叶植物之一。解析牡丹红色叶性状形成的生理机制和关键基因，可为牡丹观叶品种的选育提供理论依据。【方法】 采集长红叶期的卵叶牡丹（Paeonia qiui）和短红叶期的‘洛阳红’（P. Luoyang Hong）叶片6个发育时期样品，通过色差仪测定叶色参数，使用酶标法测定花青苷和叶绿素含量，并采用高效液相色谱（HPLC）分析花青苷组分，最后通过qPCR检测花青苷合成相关基因的表达。【结果】 表型观察与色差分析表明，卵叶牡丹叶片红度较高，红叶持续期长，可维持到开花前；而‘洛阳红’虽早期新叶呈现红色，但展叶后很快转绿。HPLC分析表明，2种牡丹的叶片均以芍药素-3,5-二葡萄糖苷（Pn3G5G）为主要花青苷成分，卵叶牡丹的总花青苷含量显著高于‘洛阳红’，尤其在S4时期差异明显。qPCR结果显示，卵叶牡丹叶片中花青苷合成相关结构基因CHS、DFR和ANS的表达水平与花青苷含量变化趋势一致，相关性分析表明，DFR表达水平与花青苷含量呈极显著正相关，ANS表达水平与花青苷含量呈显著正相关；二者作为关键结构基因影响花青苷的合成。‘洛阳红’叶片中CHS、F3H、DFR和ANS的表达水平与花青苷含量趋势基本一致，相关性分析显示，DFR与ANS表达水平均与花青苷含量呈显著正相关。【结论】 卵叶牡丹和‘洛阳红’叶片呈现红色的主要色素均为芍药素-3,5-二葡萄糖苷，DFR和ANS是牡丹叶片花青苷生物合成途径中的关键结构基因，其表达水平与花青苷积累密切相关，进而影响牡丹红色叶片的形成。

关键词: 牡丹, 卵叶牡丹, 叶色, 花青苷, 结构基因

Abstract:

【Objective】 Tree peony, renowned for its ornamental flowers, also serves as a spring foliage plant due to the striking purple-red coloration of its young leaves. Elucidating the physiological mechanisms and key genes underlying red color leaf traits provides a theoretical basis for breeding ornamental foliage cultivars. 【Method】 Leaf samples were collected at six developmental stages from Paeonia qiui and P. Luoyang Hong. Leaf color parameters were measured using a colorimeter. Anthocyanin and chlorophyll contents were quantified through enzymatic methods. The composition of anthocyanins was analyzed by high performance liquid chromatography (HPLC). Furthermore, quantitative real-time PCR (qPCR) was utilized to examine the expression levels of structural genes involved in anthocyanin biosynthesis. 【Result】 Phenotypic observations and colorimetric analyses revealed that P. qiui exhibits a higher redness and a prolonged red-leaf period that persists until flowering. Conversely, P. Luoyang Hong displays red coloration in its young leaves during the early stages, but this red hue rapidly transitions to green following leaf expansion. HPLC analysis revealed that both tree peonies had peonidin-3,5-diglucoside (Pn3G5G) as the main anthocyanin component in leaves. The total anthocyanin content in P. qiui was significantly higher than that in P. Luoyang Hong, with the most pronounced difference observed during the S4 stage. qPCR results showed that the expression levels of CHS, DFR, and ANS in P. qiui were consistent with the trend in anthocyanin content. Correlation analysis further revealed a highly significant positive correlation between DFR expression levels and anthocyanin content, as well as a significant positive correlation for ANS. These findings underscore the critical role of DFR and ANS as key structural genes in anthocyanin biosynthesis. Similarly, in P. Luoyang Hong, the expression levels of CHS, F3H, DFR, and ANS generally aligned with the trends in anthocyanin content. Correlation analysis confirmed that both DFR and ANS expression levels were significantly positively correlated with anthocyanin content. 【Conclusion】 The primary pigment responsible for the red coloration in the leaves of P. qiui and P. Luoyang Hong is peonidin-3,5-diglucoside. DFR and ANS are key structural genes in the anthocyanin biosynthesis pathway of tree peony. Their expression levels are closely associated with anthocyanin accumulation, which in turn influences the formation of red leaf coloration in peonies.

Key words: tree peony, Paeonia qiui, leaf color, anthocyanin, structural genes

魏晨曦, 董山榕, 王小满, 罗建让. 基于叶色表型和花青苷积累特征的牡丹红色叶性状解析[J]. 中国农业科学, 2025, 58(23): 5046-5056.

WEI ChenXi, DONG ShanRong, WANG XiaoMan, LUO JianRang. Analysis of Red Color Leaf Traits in Tree Peony Based on Leaf Color Phenotypes and Anthocyanin Accumulation Characteristics[J]. Scientia Agricultura Sinica, 2025, 58(23): 5046-5056.

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基因 Gene name	正向引物 Forward primer (5′-3′)	反向引物 Reverse primer (5′-3′)
PqCHS, PsCHS	TAGTCCCGGATAGCGATGGT	TGCCTGGTCGCTTGTAGTTT
PqCHI, PsCHI	GTCAGCGGAGTCAAGATTGAA	AATATCCCGGAAGAAGTCGTC
PqF3H, PsF3H	ACGAAATCCCAATCATCTCCC	CTATTTCACGCCAATCTCGCA
PqF3’H, PsF3’H	CAAGGCCGACAAATTCAAGGA	ACCCTCGCTATCAACATCATC
PqDFR, PsDFR	TGGTGGATGGAAGCTTTGATG	TAAACTCCATGTCACTCCAGC
PqANS, PsANS	CAGCATCACCAACATCTTCGA	CAGCAATCTTCCCAGTCTCTT
PqUFGT, PsUFGT	ACCATCATCCGCTACCTTTC	CACTAAACAGCTCACCTTCC

种类 Species	时期 Period	英国皇家园艺学会比色卡 RHSCC
卵叶牡丹 P. qiui	S1	红色组47B Red group 47B
	S2	红色组47A Red group 47A
	S3	灰红组181B Greyed-red group 181B
	S4	灰红组182B Greyed-red group 182B
	S5	灰红组177C Greyed-red group 177C
	S6	绿色组135C Green group 135C
洛阳红 P. Luoyang Hong	S1	红色组45C Red group 45C
	S2	灰红组180C Greyed-red group 180C
	S3	灰红组176D Greyed-red group 176D
	S4	绿色组143C Green group 143C
	S5	绿色组141C Green group 141C
	S6	绿色组139B Green group 139B