Establishment and Optimization of Cell Suspension Culture System for Vitis vinifera

doi:10.13560/j.cnki.biotech.bull.1985.2017-0075

Abstract

Abstract: In order to establish a rapid and stable cell suspension system of grapevine callus,the stems,leaves and petioles of cv. Thompson seedless and cv. Pinot Noir were used as explants. Through the selection of basic medium,the ratio and concentration of plant growth regulators and the presence or absence of PVP,the induction method of loose callus was optimized,and a stable cell suspension culture system was established. The results showed that the MS as the basic medium combined with 2.0 mg/L NAA and 0.3 mg/L 6-BA was suitable for the induction of loose callus of grapevine while the stems of cv. Thompson seedless and cv. Pinot Noir were as explants. Under the optimal culture conditions of B5-based medium supplemented with 1.0 mg/L 2,4-D,0.5 mg/L 6-BA and 0.2% PVP,a rapid and stable suspension culture system for cv. Thompson seedless was established. The growth curve of grapevine suspension culture cells was S-shaped,the logarithmic phase was on the 6-18 d after inoculation,and the cell growth reached the maximum at the 21 d. The results of both cell viability assay and TTC staining showed that the cell viability was the strongest on the 9 d after inoculation. Conclusively,a rapid and stable suspension culture system of cv. Thompson seedless is established,and it is more efficient to select 7-9 d cells with the stronger cell viability and faster growth to carry out the genetic transformation.

Key words: grapevine, loose callus, cell suspension culture, cell viability

WANG Ling, LI Yan, DAI Wei-na, YAN Jing, ZHANG Chao-hong. Establishment and Optimization of Cell Suspension Culture System for Vitis vinifera[J]. Biotechnology Bulletin, 2018, 34(8): 80-86.

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