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Author Ghedira, Rim ♦ Buck, Sylvie ♦ Ex, Frédéric ♦ Angen, Geert ♦ Depicker, Ann
Source SpringerLink
Content type Text
Publisher Springer Berlin Heidelberg
File Format PDF
Copyright Year ©2013
Language English
Subject Domain (in DDC) Natural sciences & mathematics ♦ Life sciences; biology
Subject Keyword Agrobacterium ♦ CRE recombinase ♦ Transformation efficiency ♦ Transgene expression ♦ Plant Sciences ♦ Agriculture ♦ Ecology ♦ Forestry
Abstract T-DNA transfer and integration frequencies during Agrobacterium-mediated root explant cocultivation and floral dip transformations of Arabidopsis thaliana were analyzed with and without selection for transformation-competent cells. Based on the presence or absence of CRE recombinase activity without or with the CRE T-DNA being integrated, transient expression versus stable transformation was differentiated. During root explant cocultivation, continuous light enhanced the number of plant cells competent for interaction with Agrobacterium and thus the number of transient gene expression events. However, in transformation competent plant cells, continuous light did not further enhance cotransfer or cointegration frequencies. Upon selection for root transformants expressing a first T-DNA, 43–69 % of these transformants showed cotransfer of another non-selected T-DNA in two different light regimes. However, integration of the non-selected cotransferred T-DNA occurred only in 19–46 % of these transformants, indicating that T-DNA integration in regenerating root cells limits the transformation frequencies. After floral dip transformation, transient T-DNA expression without integration could not be detected, while stable T-DNA transformation occurred in 0.5–1.3 % of the T1 seedlings. Upon selection for floral dip transformants with a first T-DNA, 8–34 % of the transformants showed cotransfer of the other non-selected T-DNA and in 93–100 % of them, the T-DNA was also integrated. Therefore, a productive interaction between the agrobacteria and the female gametophyte, rather than the T-DNA integration process, restricts the floral dip transformation frequencies.
ISSN 00320935
Age Range 18 to 22 years ♦ above 22 year
Educational Use Research
Education Level UG and PG
Learning Resource Type Article
Publisher Date 2013-08-24
Publisher Place Berlin/Heidelberg
e-ISSN 14322048
Journal Planta
Volume Number 238
Issue Number 6
Page Count 13
Starting Page 1025
Ending Page 1037

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Source: SpringerLink