目的 查耳酮合成酶（chalcone synthase，CHS）是类黄酮物质合成的首个关键酶，在植物类黄酮物质积累及逆境应答过程中起着重要作用。旨在鉴定掌叶覆盆子CHS基因家族成员，明确其在不同组织器官中及果实成熟中的表达模式。方法 通过生物信息学方法鉴定掌叶覆盆子CHS家族成员，分析了它们的理化性质、染色体定位、系统进化树、保守基序、基因结构和顺式作用元件等，同时通过荧光定量PCR检测它们在果实不同发育时期及不同组织中的表达量。结果 掌叶覆盆子有15个CHS家族成员，分为7个亚族，不均匀地分布在4条染色体上。这些RcCHS蛋白的长度蛋白数介于361～541，都定位于细胞质，且存在1对串联复制基因。同一亚族成员基因结构、保守基序和蛋白质二级结构相类似。RcCHS家族基因的启动子区域存在大量生长发育、激素和环境胁迫应答相关的调控元件。RcCHS家族基因在掌叶覆盆子不同组织中均有表达，并具有明显的组织特异性，此外在掌叶覆盆子果实不同成熟阶段和100 μmol/L茉莉酸甲酯（methyl jasmonate，MeJA）处理下的表达量均存在显著差异。发现RcCHS11在果实中高丰度表达，并受到MeJA强烈诱导，与类黄酮物质在掌叶覆盆子果实中的积累规律相一致，推测是类黄酮成分合成途径上的关键酶。结论 在全基因组上鉴别出15个掌叶覆盆子RcCHS家族成员，它们的表达呈现器官特异性，并在果实不同时期呈现差异表达，RcCHS11与果实中类黄酮积累呈现正向响应。

Objective Chalcone synthase (CHS) is the first key enzyme for flavonoids biosynthesis, and plays an important role in the accumulation of flavonoids and stress response. To identify the members of CHS gene family in Zhangye Fupenzi (Rubus chingii), and define their expression patterns in different tissues (roots, stems, leaves, flowers, and fruits) and at different growth stages of fruits of R. chingii. Methods Multiple bioinformatics methods were employed to identify the CHS family members in R. chingii genome. Their physicochemical properties, chromosomal localization, phylogenetic tree, conserved motifs, gene structure, and cis-acting elements were analyzed. Simultaneously, their expression profiles in different tissues (roots, stems, leaves, flowers, and fruits) and during different stages (green fruits, green to yellow fruits, yellow fruits, and red fruits) of fruit development were detected using the real-time fluorescence quantitative polymerase chain reaction (PCR).Results A total of 15R. chingii CHS family members were identified, which were divided into 7 subfamilies, unevenly located on four chromosomes. They encoded 361—541 proteins, located in the cytoplasm, and harbored one pair of tandem replicated genes. The gene structure, conserved motifs, and secondary structure of RcCHS members within the same subfamily were similar. The promoter region of RcCHS family genes contains a large number of regulatory elements related to growth and development, hormone and environmental stress response. RcCHS family genes were expressed in different tissues (roots, stems, leaves, flowers, and fruits) with obvious tissue specificity. In addition, the expression levels of RcCHS family gene were significantly different at different fruit development stages (green fruits, green to yellow fruits, yellow fruits, and red fruits) and under 100 μmol/L methyl jasmonate (MeJA) treatment. Among them, RcCHS11 was highly expressed in fruits ofR. chingii, and induced strongly by MeJA treatment, which was coincided with the accumulation of flavonoids, suggesting that RcCHS11 might be a critical enzyme involved in the biosynthesis of flavonoids. Conclusion A total of 15 members of RcCHS family were systematically identified from R. chingii at the genome level. They exhibited tissue specific expression profiles, and possessed different responses at different development stages. RcCHS11 showed positively response to the flavonoids accumulation in fruit.

R286.12

浙江省“十四五”育种专项中药材协作组项目（2021C02074-1）;浙江省基础公益研究计划项目（LY23H280003）;浙江中医药大学科研项目（2022JKZKTS15）;三峡库区道地药材开发利用重庆市重点实验室开放课题项目（KFKT2022013）