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中国农学通报 ›› 2022, Vol. 38 ›› Issue (24): 92-99.doi: 10.11924/j.issn.1000-6850.casb2022-0109

所属专题: 生物技术 园艺

• 生物科学 • 上一篇    下一篇

西番莲细胞悬浮系建立与培养条件优化

邝瑞彬(), 杨敏, 周陈平, 杨护, 黄炳雄, 魏岳荣()   

  1. 广东省农业科学院果树研究所/农业农村部南亚热带果树生物学与遗传资源利用重点实验室/广东省热带亚热带果树研究重点实验室,广州 510640
  • 收稿日期:2022-03-02 修回日期:2022-05-02 出版日期:2022-08-25 发布日期:2022-08-22
  • 通讯作者: 魏岳荣
  • 作者简介:邝瑞彬,女,1978年出生,广东广州人,副研究员,博士,研究方向:果树育种。通信地址:510640 广州天河区五山大丰二街80号,Tel:020-38765468,E-mail: rbkuang15@163.com
  • 基金资助:
    广东省农业农村厅“广东省现代农业产业技术体系创新团队建设项目(优稀水果产业)”项目(2021KJ116);广东省农业农村厅乡村振兴战略专项资金(构建现代农业体系)项目“农作物种质资源库(圃)建设与资源收集保存、鉴评”(粤农计[2018]36号)

Cell Suspension Culture System for Passion Fruit: Establishment and Optimization

KUANG Ruibin(), YANG Min, ZHOU Chenping, YANG Hu, HUANG Bingxiong, WEI Yuerong()   

  1. Institute of Fruit Tree Research, Guangdong Academy of Agricultural Sciences/ Key Laboratory of South Subtropical Fruit Biology and Genetic Resource Utilization, Ministry of Agriculture and Rural Affairs/ Guangdong Provincial Key Laboratory of Tropical and Subtropical Fruit Tree Research, Guangzhou 510640
  • Received:2022-03-02 Revised:2022-05-02 Online:2022-08-25 Published:2022-08-22
  • Contact: WEI Yuerong

摘要:

本研究旨在建立西番莲细胞悬浮培养体系,并优化悬浮系培养条件,为西番莲遗传工程提供基础。本研究以西番莲无菌苗为材料,研究西番莲愈伤组织的诱导和建立细胞悬浮系,并结合单因素试验与正交试验优化悬浮系培养条件。结果表明,西番莲愈伤诱导的外植体最佳选择是幼苗胚轴,愈伤诱导激素选择2 mg/L 2,4-D为宜,出愈率达83.3%。以疏松健康的愈伤组织培养细胞悬浮系,经单因素试验和正交试验,细胞悬浮系培养最佳培养培养基为MS培养基+2 mg/L 2,4-D+0.2 g/L Glutamine+30 g/L蔗糖,培养基pH 6.1,起始细胞用量为6.0 mg/mL,摇床转速为100 r/min,28℃暗培养,此条件下建立的细胞悬浮系均匀一致,细胞生长旺盛,西番莲悬浮细胞生长曲线呈“S”型,继代最佳时期为12~18天。本研究为后续利用细胞悬浮系建立西番莲高效细胞工程及遗传转化技术体系提供良好的材料和技术基础,对加强西番莲种业发展具重要意义。

关键词: 西番莲, 细胞悬浮系, 愈伤组织, 诱导, 生长曲线

Abstract:

The aim of this study is to establish a rapid and stable cell suspension culture system for passion fruit callus, and to optimize the culture conditions, which may lay a foundation for genetic engineering of passion fruit. The seedlings of passion fruit were used as plant materials and their different parts were used as explants to induce callus, which were then used to develop cell suspension culture. Single factor and orthogonal experiment were carried out to optimize the cell suspension culture conditions. Results showed that the basic medium MS with 2 mg/L 2,4-D was suitable for the induction of callus of passion fruit while the seedling hypocotyl was suitable as explants,with a callus induction rate of 83.3%. Under the optimal culture condition of MS-based medium supplemented with 2 mg/L 2,4-D, 0.2 g/L Glutamine and 30 g/L sucrose, and pH of 6.1, a rapid and stable suspension culture system for passion fruit was established. The initial cell dosage was 6.0 mg/mL and shaker speed was 100 r/min, and 28℃ dark culture was adopted. Under the above growth condition, the suspension cells grew uniformly and vigorously. The growth curve of the suspension cells was S-shaped,and the logarithmic phase was on the 12-18 d after inoculation and the cells were cultured under suitable conditions. This study lays a material and technology base for cell engineering breeding and genetic transformation system of passion fruit, which is important for the development of passion fruit industry.

Key words: passion fruit, cell suspension culture, callus, induction, growth curve

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