葡萄细胞悬浮培养体系的建立和优化

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

生物技术通报 ›› 2018, Vol. 34 ›› Issue (8): 80-86.doi: 10.13560/j.cnki.biotech.bull.1985.2017-0075

葡萄细胞悬浮培养体系的建立和优化

王玲¹, 李琰², 代伟娜¹, 严静¹, 张朝红¹

1. 西北农林科技大学园艺学院农业部西北地区园艺作物生物学与种质创制重点实验室,杨凌 712100;
2. 西北农林科技大学生命科学学院,杨凌 712100

收稿日期:2018-01-18 出版日期:2018-08-26 发布日期:2018-09-04
作者简介:王玲,女,硕士研究生,研究方向：果树种质资源与育种;E-mail：lingwang0922@163.com
基金资助:
陕西省重点研发计划项目（2017NY-026）,国家现代农业产业技术体系建设专项（CARS-30-yz-7）

Establishment and Optimization of Cell Suspension Culture System for Vitis vinifera

WANG Ling¹, LI Yan², DAI Wei-na¹, YAN Jing¹, ZHANG Chao-hong¹

1. College of Horticulture,Northwest A&F University,Key Laboratory of Biology and Genetic Improvement of Horticultural Crops（Northwest Region）,Ministry of Agriculture,Yangling 712100;
2. College of Life Science,Northwest A＆F University,Yangling 712100

Received:2018-01-18 Published:2018-08-26 Online:2018-09-04

摘要/Abstract

摘要： 为了利用葡萄愈伤组织建立快速稳定的葡萄细胞悬浮体系,以无核白和黑比诺葡萄无菌幼苗的茎段、叶片以及叶柄为外植体,通过基本培养基、不同植物生长调节剂配比及其浓度、PVP等优化疏松型愈伤组织的诱导方法并建立稳定的细胞悬浮培养体系。结果表明,以无核白和黑比诺葡萄茎段为外植体,在添加2.0 mg/L NAA和0.3 mg/L 6-BA的MS培养基上适合疏松型愈伤组织的诱导。以B5为基本培养基添加1.0 mg/L 2,4-D和0.5 mg/L 6-BA 以及0.2% PVP的优化培养条件下,建立了快速稳定的无核白葡萄悬浮培养体系。葡萄悬浮培养细胞生长曲线呈S形,接种后6-18 d处于对数生长期,第21 天细胞增长量达到最大值。细胞活力测定与TTC染色结果一致表明,接种后培养9 d细胞活力最强。建立了快速稳定的无核白葡萄悬浮培养体系,选择细胞活力最强且细胞快速增值的对数期7-9 d的细胞进行葡萄悬浮细胞遗传转化,效率较高。

关键词: 葡萄苗, 疏松型愈伤组织, 细胞悬浮培养, 细胞活力^?

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

王玲, 李琰, 代伟娜, 严静, 张朝红. 葡萄细胞悬浮培养体系的建立和优化[J]. 生物技术通报, 2018, 34(8): 80-86.

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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葡萄细胞悬浮培养体系的建立和优化

Establishment and Optimization of Cell Suspension Culture System for Vitis vinifera

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