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Stable genetic transformation of Arabidopsis thaliana by Agrobacterium inoculation in planta
Author(s) -
Chang Seok So,
Park Soon Ki,
Kim Byung Chul,
Kang Bong Joong,
Kim Dal Ung,
Nam Hong Gil
Publication year - 1994
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.1994.05040551.x
Subject(s) - agrobacterium , arabidopsis thaliana , transformation (genetics) , biology , inoculation , arabidopsis , botany , genetics , gene , horticulture , mutant
Summary Stable genetic transformation of Arabidopsis thaliana was achieved by simple in planta inoculation of Agrobacterium tumefaciens strain LBA4404 harboring a binary vector pBl121. The transformation procedure, which we call in planta transformation, involves severing of apical shoots at their bases, inoculation with Agrobacterium at the severed sites, and in planta generation of shoots from the severed sites. On average, 5.5% of the newly formed shoots produced transformed progenies. These progenies (T 2 generation) contained T‐DNA in the genome as examined by assaying the T‐DNA encoded β‐glucuronidase and kanamycin resistance and by genomic Southern blot analysis, the copy number of the T‐DNAs in the Arabidopsis genome being single (33%) or multiple. The genetic behavior of the transformants examined at the T 3 and T 4 generations or with the F 2 progenies of the outcrosses between transformants and wild‐type plants showed that most of the inserted T‐DNA are inherited in a Mendelian fashion. This procedure provides a new approach for simple and efficient transformation of A. thaliana , obviating the need for plant regeneration from tissue explants in vitro .