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菊花品种‘万代风光’再生及遗传转化体系的建立

  • 李晶晶 ,
  • 李艳飞 ,
  • 王安琪 ,
  • 王佳颖 ,
  • 邓成燕 ,
  • 卢敏 ,
  • 马剑英 ,
  • 戴思兰
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  • 北京林业大学园林学院,城乡生态环境北京实验室,花卉种质创新与分子育种北京市重点实验室,国家花卉工程技术研究中心,北京 100083

收稿日期: 2024-10-10

  修回日期: 2025-01-03

  网络出版日期: 2025-01-22

基金资助

国家自然科学基金项目(No.32171849, No.32371948)和福建省科技计划项目(No.2022S0004).

Establishment of Regeneration and Genetic Transformation System for Chrysanthemum Cultivar ‘Wandai Fengguang’

  • LI Jing-Jing ,
  • LI Yan-Fei ,
  • YU An-Qi ,
  • YU Jia-Ying ,
  • DENG Cheng-Yan ,
  • LV Min ,
  • MA Jian-Yang ,
  • DAI Sai-Lan
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  • National Engineering Research Center for Floriculture, Beijing Key Laboratory of Ornamental Plants Germplasm Innovation & Molecular Breeding, Beijing Laboratory of Urban and Rural Ecological Environment, School of Landscape Architecture, Beijing Forestry University, Beijing 100083, China

Received date: 2024-10-10

  Revised date: 2025-01-03

  Online published: 2025-01-22

摘要

菊花品种‘万代风光’(Chrysanthemum ×morifolium ‘Wandai Fengguang’)的色素背景适宜利用分子育种技术调控花瓣铁离子浓度而培育蓝色花,且其在夏秋两季均可开花,也是研究菊花开花期分子调控机理的重要材料,但缺少高效的再生体系和遗传转化体系。本研究以该品种为试验材料,研究不同外植体类型和不同植物生长调节剂组合对其再生的影响,并探究农杆菌介导的遗传转化方法中相关因素对遗传转化效率的影响。研究结果表明:适宜‘万代风光’再生的最适外植体为茎间薄层,最适培养基为MS+1.5 mg·L-1 6-BA+0.6 mg·L-1 NAA,分化率为70.06%,不定芽生成系数为3.37;经过试验确定:茎间薄层分化和不定芽生根的卡那霉素选择压分别为7.5 mg·L-1和5.0 mg·L-1。预培养1天,OD600=0.8,处理5分钟,黑暗共培养3天为遗传转化处理的最佳体系。经过卡那霉素筛选共获得抗性苗15株,PCR鉴定发现2株阳性苗,转化效率为13.33%。本研究为利用这一独特品种资源解析菊花基因功能和进行定向改良的分子育种奠定了基础,为其它菊花品种的再生和转化体系的建立提供了参考。

本文引用格式

李晶晶 , 李艳飞 , 王安琪 , 王佳颖 , 邓成燕 , 卢敏 , 马剑英 , 戴思兰 . 菊花品种‘万代风光’再生及遗传转化体系的建立[J]. 植物学报, 0 : 1 -0 . DOI: 10.11983/CBB24151

Abstract

The pigment background of the chrysanthemum cultivar 'Wandai Fengguang' is suitable for the cultivation of blue flowers by using molecular breeding techniques to regulate the concentration of iron ions in petals. While it can bloom in both summer and autumn, it is also an important material for studying the molecular regulation mechanism of chrysanthemum flowering period. However, it lacks an efficient regeneration and genetic transformation system. In this study, this variety was used as the experimental material to study the effects of different explant types and different combinations of plant growth regulators on its regeneration, and to investigate the effects of relevant factors on the efficiency of genetic transformation in the agrobacterium-mediated genetic transformation method. It was experimentally determined that the most suitable explant for the regeneration of 'Wandai Fengguang' was the transverse thin cell layers, and the optimal culture medium was MS+1.5 mg·L-1 6-BA+0.6 mg·L-1 NAA, with a differentiation rate of 70.06% and an adventitious shoot coefficient of 3.37. The kanamycin selection pressures for the differentiation of the transverse thin cell layers and the adventitious bud rooting were 7.5 mg·L-1 and 5.0 mg·L-1, respectively. After experimentation, the optimal system for genetic transformation treatment was determined to be pre-culture for 1 day, OD600=0.8, treatment for 5 minutes, and co-culture in the dark for 3 days. Fifteen resistant plantlets were screened on kanamycin medium, and two positive plantlets were revealed by PCR identification, with a transformation efficiency of 13.33%. This study laid the foundation for the gene function analysis and targeted improvement molecular breeding of chrysanthemum by using this kind of unique variety resource, and provided reference for the establishment of regeneration and transformation system for other chrysanthemum varieties.
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