Efficient Agrobacterium-Mediated Transformation of the Liverwort Marchantia polymorpha Using Regenerating Thalli

被引:218
作者
Kubota, Akane [1 ]
Ishizaki, Kimitsune [1 ]
Hosaka, Masashi [1 ]
Kohchi, Takayuki [1 ]
机构
[1] Kyoto Univ, Grad Sch Biostudies, Kyoto 6068502, Japan
基金
日本学术振兴会;
关键词
Agrobacterium-mediated transformation; bryophyte; evolution; gametophyte; T-DNA; GENETIC-TRANSFORMATION; COMPLETE SEQUENCE; PLANTS; ORGANIZATION; BIOLOGY; DNA; L; GENOMICS; CULTURE; SYSTEM;
D O I
10.1271/bbb.120700
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The thallus, the gametophyte body of the liverwort Marchantia polymorpha, develops clonal progenies called gemmae that are useful in the isolation and propagation of isogenic plants. Developmental timing is critical to Agrobacterium-mediated transformation, and high transformation efficiency has been achieved only with sporelings. Here we report an Agrobacterium-mediated transformation system for M. polymorpha using regenerating thalli. Thallus regeneration was induced by cutting the mature thallus across the apical-basal axis and incubating the basal portion of the thallus for 3d. Regenerating thalli were infected with Agrobacterium carrying binary vector that contained a selection marker, the hygromycin phosphotransferase gene, and hygromycin-resistant transformants were obtained with an efficiency of over 60%. Southern blot analysis verified random integration of 1 to 4 copies of the T-DNA into the M. polymorpha genome. This Agrobacterium-mediated transformation system for M. polymorpha should provide opportunities to perform genetic transformation without preparing spores and to generate a sufficient number of transformants with isogenic background.
引用
收藏
页码:167 / 172
页数:6
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