阳春砂（Amomum villosum Lour.）是传统中药材砂仁的主要基原植物之一，具 有重要的药用价值，但其长达2-3年的营养生长期严重制约了砂仁药材的生产效率。 所以，选育早花早实品种并解析花期调控机制成为缩短砂仁的生产周期、提高砂仁 产量的关键突破口。开花是植物从营养生长向生殖生长转变的关键节点，其调控机 理一直是植物分子生物学领域的研究热点。在阳春砂育种中，早花早实作为关键目 标性状，不仅能缩短生长周期、促进果实早熟，还能通过集中阳春砂的花期提高昆 虫对花的传粉效率，进而实现砂仁药材的产量增加。 目前，阳春砂开花进程及花器官形成的分子机制尚未明晰。鉴于水稻（Oryza sativa L.）中的 Ghd7 基因（grain number，plant height and heading date7），能够显 著延迟开花时间，这为研究阳春砂开花调控提供了重要线索。本研究选取课题组前 期筛选到的阳春砂开花相关差异表达基因 Unigene17286（注释为 CO，后命名为 AvGhd7）作为目的基因，通过基因克隆、生物信息学分析、基因表达模式分析、过 表达载体构建、烟草亚细胞定位及拟南芥异源表达等实验方法，初步探究了该基因 在阳春砂花期调控中的作用机制。 主要研究结果如下： 1. 首次在阳春砂中克隆了成花相关的基因AvGhd7，其 CDS全长为693 bp，编 码230个氨基酸，含有CONSTANS（CO）基因典型的CCT结构域，属于CCT超基 因家族。系统进化分析表明，AvGhd7与姜科植物姜的同源蛋白聚为一类，序列百分 比一致性达85.17%。 2. 通过qRT-PCR检测发现，AvGhd7基因在开花部位（花芽）表达量显著低于 其他组织部位（叶片和匍匐茎尖），在早实单株中的表达量低于非早实单株，表明 该基因可能作为负调控因子参与阳春砂开花进程。 3. 利用烟草叶片瞬时表达系统进行亚细胞定位，结果显示AvGhd7蛋白定位于 细胞膜。通过拟南芥异源表达实验，初步筛选获得T0代阳性植株，为后续功能验证 奠定了基础。 4. 为探究AvGhd7 在阳春砂及其近缘种间的遗传多态性，采用同一PCR特异 性引物对豆蔻属7份不同种质（海南砂、老挝砂、巴色砂、红壳砂、绿壳砂、红刺 绿壳砂及阳春砂）及阳春砂的45份不同种质间进行PCR扩增、测序和序列比对。 结果显示豆蔻属7份不同种质的序列差异显著，STS分子标记引物可有效区分不同 种质。系统进化分析结果进一步表明，阳春砂与红刺绿壳砂的亲缘关系最近，与老 挝砂的亲缘关系最远。 

本研究首次筛选并克隆到调控阳春砂与成花相关的候选负调控基因 AvGhd7， 并初步验证了其功能，为多年生药用植物阳春砂的早花早实品种培育提供了关键分 子靶点和理论支持，具有重要的应用前景。 

Amomum villosum Lour., the main source variety of the traditional Chinese medicine Sharen, has significant medicinal value. However, its long vegetative growth period of 2 3 years severely restricts production efficiency. Breeding early-flowering and early fruiting varieties and elucidating the mechanism of flowering regulation have become key breakthroughs for shortening the production cycle and increasing yield. Flowering is a critical transition from vegetative to reproductive growth in plants, and its regulatory mechanism has always been a research hotspot in plant molecular biology. In the breeding of A. villosum, early-flowering and early-fruiting traits, as the core target traits, not only shorten the growth cycle and promote early fruiting but also increase pollination efficiency by concentrating the flowering period, thereby achieving yield gains. Currently, the molecular mechanism of flowering and flower organ formation in A. villosum remains unclear. Given that the Ghd7 gene in rice (Oryza sativa L.) can significantly delay flowering time, this provides an important clue for studying the flowering regulation of A. villosum. In this study, the flowering-related differentially expressed gene Unigene17286 (annotated as CO and later named AvGhd7) was selected as the target gene from the previous screening by the research group. Through gene cloning, expression pattern analysis, overexpression vector construction, and Arabidopsis thaliana heterologous transformation experiments, the role of this gene in the flowering regulation of A. villosum was systematically explored. The main research results are as follows: 1. The flowering-related gene AvGhd7 was cloned for the first time in A. villosum. Its CDS is 693 bp in length, encoding 230 amino acids, and contains the typical CCT domain of the CONSTANS (CO) gene, belonging to the CCT supergene family. Phylogenetic analysis indicated that AvGhd7 clustered with the homologous protein of Zingiber officinale in the Zingiberaceae family, with a sequence identity of 85.17%. 2. qPCR detection revealed that the expression level of AvGhd7 in the flowering site (flower buds) was significantly lower than in other tissues (leaves and stolons), and the expression level in early-fruiting individuals was lower than in non-early-fruiting individuals, suggesting that this gene may act as a negative regulator in the flowering process of A. villosum. 3. Subcellular localization was performed using the tobacco leaf transient expression system, and the results showed that the AvGhd7 protein was located on the cell membrane. Through Arabidopsis heterologous transformation experiments, T0 generation positive plants were initially obtained, laying the foundation for subsequent functional verification. 4. To explore the genetic polymorphism of AvGhd7 among A. villosum and its related species, PCR amplification, sequencing, and sequence alignment were conducted using the same PCR primers on seven different germplasms of the genus Amomum (Hainan sha, Laos sha, Pakse sha, red-shelled sha, green-shelled sha, red-spined green-shelled sha, and A. villosum). The results showed significant sequence differences among species, and the STS molecular marker primers could effectively distinguish each species. Evolutionary analysis further indicated that A. villosum was most closely related to red-spined green shelled sha and most distantly related to Laos sha. This study identified the candidate gene AvGhd7 regulating the flowering time of A. villosum for the first time and preliminarily indicated its function, providing a key molecular target and theoretical support for the breeding of early-flowering and early fruiting varieties of the perennial medicinal plant A. villosum, with significant application prospects.