枸杞Lb13b基因的表达分析及转烟草研究植物遗传资源学报.PDF

植物遗传资源学报 2018，19 Journal of Plant Genetic Resources DOI:10.13430/ki.jpgr.20180324001 枸杞Lb14-3-3b基因的表达分析及转烟草研究1 徐 蕊，周 丽，李瑞国，岳思君，毛桂莲，郑 蕊 （宁夏大学生命科学学院西部特色生物资源保护与利用教育部重点实验室，银川 750021） 摘要：在宁夏枸杞 （Lycium barbarumL.）品种宁杞 1号花药发育差异蛋白质组学研究的基础上，克隆了一个花药发育相 关基因Lb14-3-3b，证实该基因在花器官中优势表达。本试验进一步利用荧光定量PCR 技术分析枸杞Lb14-3-3b基因在花药发 育过程中不同时期的表达特征，并通过构建植物过表达载体Lb14-3-3b-pCambia1305.1-35s，经根瘤农杆菌介导法转化模式植 物烟草，探究该基因在植物生长发育中的功能。结果表明，在枸杞花药发育的各个时期，Lb14-3-3b都有表达，在花药二核花 粉粒时期表达量最高。与野生型烟草相比，转Lb14-3-3b基因烟草生长发育迟缓，植株矮小，花器官畸变，花瓣缺少致使花 型趋于四角化，雄蕊及花丝数目缺少且发育异常。推测枸杞Lb14-3-3b基因在烟草生长发育及花器官发育中起调控作用，研 究结果为进一步探讨该基因在枸杞生长发育中的调控作用提供参考依据。 关键词：枸杞；Lb14-3-3b；烟草; 遗传转化 TranscriptionalAnalysisoftheGeneLb14-3-3b inWolfberry and Functional CharacterizationoftheTransgenicLinesinTobacco XURui ，ZHOU Li，LIRui-guo，YUE Si-jun，MAO Gui-lian，ZHENGRui （WesternBiologicalResourcesProtection and UtilizationLab of NationalEducationMinistry, College of LifeScience,Ningxia University, Yinchuan 750021） Abstract:By analyzing the proteome datasets during the anther development in wolfberry variety ‘Ning QiNo.1’, we identified and isolated an anther development related gene Lb14-3-3b that is observed in floral organs. In order to further understand its transcriptional characteristics in floral organs, we determined by real-time quantitative PCR approach the transcriptional level of Lb14-3-3b at different stages of wolfberry anther development. In addition, we transformed this gene into model plant tobacco via Agrobacterium tumefaciens mediated transformation, driven by the constitutive 35S promoter. The results showed that Lb14-3-3b expressed at all stages of the wolfberry anther development, particularly at the stage of two-nuclear pollen grain with the highest expression level. Compared with wild type, transgenic tobacco plants grew delayed growth and dwarfed phenotype, floral organ distor