目的 解析枸杞Lycium barbarum LbbHLH94-like基因的表达特征及其在类胡萝卜素合成中的调控功能，为揭示枸杞果实品质形成的分子机制提供理论依据。方法 以宁夏枸杞‘宁杞1号’为材料，采用PCR、qRT-PCR等技术克隆并分析LbbHLH94-like在不同组织、果实发育阶段、非生物胁迫及外源激素处理下的表达模式；构建过表达载体转化枸杞，分析转基因植株类胡萝卜素代谢关键基因表达、酶活性及总类胡萝卜素含量。结果 成功克隆LbbHLH94-like基因，其开放阅读框全长888 bp，编码296个氨基酸，序列与基因组参考序列一致；该基因具有明显时空表达特异性，在果实成熟期表达量最高，并响应高温、低温、盐、干旱胁迫及6-苄氨基嘌呤（6-benzylaminopurine，6-BA）、吲哚-3-丁酸（indole-3-butyric acid，IBA）、赤霉素（gibberellic acid，GA₃）诱导，外源激素整体呈低浓度促进、高浓度抑制模式；过表达LbbHLH94-like可显著上调PDS基因表达、提高PSY与PDS酶活性，抑制类胡萝卜素裂解双加氧酶4（carotenoid cleavage dioxygenase 4，CCD4）基因表达及酶活性，显著提升果实总类胡萝卜素含量。结论 LbbHLH94-like是枸杞类胡萝卜素合成的关键正向调控因子，同时参与非生物胁迫与激素信号应答，研究结果为枸杞品质改良与精准分子育种提供重要基因资源。

Objective This study aims to clone the LbbHLH94-like gene from Gouqi (Lycium barbarum), analyze its spatiotemporal expression, stress and hormone response patterns, and clarify its regulatory function in carotenoid biosynthesis, so as to provide a theoretical basis for revealing the molecular mechanism of goji berry fruit quality formation. Methods Using Lycium barbarum cv. ‘Ningqi 1’ as material, we employed techniques such as PCR and qRT-PCR to clone and analyze the expression patterns of LbbHLH94-like in different tissues, fruit developmental stages, abiotic stresses and exogenous hormone treatments. The overexpression vector was constructed and transformed into goji plants, and the expression of key genes, enzyme activities and total carotenoid content in carotenoid metabolism were determined. Results The LbbHLH94-like gene was successfully cloned, with an 888 bp open reading frame encoding 296 amino acids, and the sequence was identical to the genome reference sequence. This gene showed obvious spatiotemporal specificity, with the highest expression level at fruit ripening stage. It responded to high and low temperature, salt, drought stresses as well as 6-BA, IBA and GA₃ induction, showing a general pattern of promotion at low concentration and inhibition at high concentration. Overexpression of LbbHLH94-like significantly up-regulated the expression of PDS, enhanced the enzyme activities of PSY and PDS, inhibited the expression and enzyme activity of CCD4, and remarkably increased total carotenoid content in fruits. Conclusion LbbHLH94-like is a key positive regulator of carotenoid biosynthesis in goji berry, and also participates in abiotic stress response and hormone signaling. The results provide important gene resources and theoretical support for quality improvement and precise molecular breeding of Lycium barbarum.

R282.12

国家自然科学基金资助项目（32460098）；宁夏回族自治区重点研发计划项目（2025BBF02004）