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园艺学报 ›› 2024, Vol. 51 ›› Issue (2): 372-384.doi: 10.16420/j.issn.0513-353x.2023-0715

• 新技术与新方法 • 上一篇    下一篇

番茄细胞分裂素与生长素响应信号可视化荧光分析

赵晓冰1,孙可一1,2,樊俊苗1,3,丁 静1,*   

  1. 1南京农业大学园艺学院,作物遗传与种质创新利用全国重点实验室,南京 210095;2北京农学院植物科学技术学院,北京 100096;3上海交通大学农业与生物学院,上海 200240
  • 出版日期:2024-02-25 发布日期:2024-02-26
  • 基金资助:
    国家自然科学基金项目(31972371);中央高校基本科研业务费专项(YDZX2023019);江苏高校优势学科建设工程项目

Visible Fluorescence Analysis of Cytokinin and Auxin Responses in Tomato

ZHAO Xiaobing1,SUN Keyi1,2,FAN Junmiao1,3,and DING Jing1,*   

  1. 1The State Key Laboratory of Crop Genetics & Germplasm Enhancement and Utilization,College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China;2College of Plant Science and Technology,Beijing University of Agriculture,Beijing 100096,China;3School of Agriculture and Biology,Shanghai Jiao Tong University,Shanghai 200240,China
  • Published:2024-02-25 Online:2024-02-26

摘要: 解析细胞分裂素和生长素的响应信号在番茄体内的动态分布,对从细胞水平阐明这两类激素的调控功能及作用机理具有重要意义。利用细胞分裂素响应启动子TCSn1驱动绿色荧光报告基因TCSn1::GFP(GFP)和生长素响应启动子DR5v2驱动红色荧光报告基因DR5v2::tdTomato(tdTomato),实时原位分析了番茄根、茎、叶、花和果实中细胞分裂素和生长素的响应信号,发现两种荧光在发芽3 ~ 7 d幼苗根冠的侧根冠、小柱起始细胞和小柱细胞及根伸长区的原形成层、基本分生组织和原表皮层,发芽5 ~ 10 d的茎尖分生组织,定植23和37 d植株茎段的维管束组织,定植32 d植株的幼嫩叶片,开花当天的花瓣和果梗,开花2 ~ 6 d果实的胚珠、胎座、小柱及表皮等组织/器官中均有分布。两种荧光的强度均在茎尖分生组织和果实胚珠中最高,并且具有类似的变化趋势:在发芽3 ~ 7 d的根冠和伸长区中先下降后有所恢复,茎段中37 d弱于23 d,发芽5 ~ 10 d茎尖分生组织和开花0 ~ 6 d果实中持续上升。本研究为番茄细胞分裂素和生长素响应分析提供了一个灵敏而稳定的可视化工具,并解析了两种激素在番茄不同器官中的分布与强度。

关键词: Solanum lycopersicum, cytokinin response, auxin response, visible fluorescence, microimaging

Abstract: Analysis of dynamic distributions of cytokinin and auxin response signals in tomato plants is of great significance for clarifying the regulatory roles and functional mechanisms of these two hormones at the cytological level. A green fluorescent reporter gene TCSn1::GFP(GFP)with a cytokinin-responsive promoter TCSn1,and a red fluorescent reporter gene DR5v2::tdTomato(tdTomato)with an auxin-responsive promoter DR5v2,were used to perform real-time in-situ analyses of the signals of cytokinin and auxin responses in tomato roots,stems,leaves,flowers,and fruits. It is found that the two types of fluorescence distributed in the tissues/organs including:lateral root cap and columella in root cap,as well as the procambium,ground meristerm and protoderm in root elongation zone at three to seven days after germination,the shoot apical meristem at five to ten days after germination,vascular tissues in stems of plants at 23 and 37 days after planting,young leaves of plants at 32 days after planting,petals and fruit peduncles at anthesis,ovules,the placenta,columella and pericarp of fruits at two to six days after anthesis. Intensities of the two types of fluorescence were both highest in the shoot apical meristem and ovules in fruits,and had a similar variation trend:first decreased and then recovered in the root cap and elongation zone during three to seven days of germination;were weaker in stem segments at 37 days than 23 days after planting;continuously rose in the shoot apical meristem at five to ten days after germination and in the fruits at zero to six days after anthesis. This study provides a sensitive and robust visualization tool for analyzing cytokinin and auxin responses in tomato,and has analyzed their distribution and intensity in different organs of tomato.

Key words: 番茄, 细胞分裂素响应, 生长素响应, 可视化荧光, 显微成像

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