期刊
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
卷 77, 期 1, 页码 167-172出版社
TAYLOR & FRANCIS LTD
DOI: 10.1271/bbb.120700
关键词
Agrobacterium-mediated transformation; bryophyte; evolution; gametophyte; T-DNA
类别
资金
- Ministry of Education, Culture, Sports, Science and Technology of Japan [23012025, 23120516, 23119510]
- Japan Society for the Promotion of Science [24580140, 24658095, 24-6987]
- Asahi Glass Foundation
- Grants-in-Aid for Scientific Research [12J06987, 24580140, 24658095] Funding Source: KAKEN
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.
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