西番莲细胞悬浮系建立与培养条件优化

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。

Cell Suspension Culture System for Passion Fruit: Establishment and Optimization

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.