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Agrobacterium-mediated genetic transformation of cereal crops

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Nadolska-Orczyk A., Przetakiewicz A., Orczyk W.
Biotechnologia
|
2000
|
issue 4
93-98
EN
The results published in recent years proved that Agrobacterium based system for genetic transformation was also suitable for cereal crops. Several groups were able to obtain transgenic rice, corn, wheat and barley using hipervirulent strains Agl1 and EHA101 (or EHA105) or 'regular' LBA4404 strain with superbinary vector pTOK233. The first phase of our research was designed to establish transformation susceptibility of two wheat, two barley and one triticale cultivars using three different bacterial systems. Two of those systems were based on hipervirulent strains: Agl1 (pDM805) and EHA101 (pGAH). The third one combined strong virulence of pTOK233 vector and commonly-used LBA 4404 strain. Putative transgenic plants (regenerated and rooted under selective pressure of appropriate factor and further confirmed with GUS or PCR) were obtained for barley (cultivar Scarlett), wheat (Torka and Kontesa) and triticale (Wanad) with Agrobacterium strain Agl1. Kontesa's putative transgenics were also obtained with the strain EHA101. The highest rate of selection of putative transgenics was for Agl1 / phosphinothricine and ranged from 9 to15% for wheat cultivars. The lowest rate for the same strain and selection was 0,5% for barley cv. Scarlett.Inoculation of 700 immature embryos of barley cv. Lot with three bacterial systems (strains, vectors and selection factors) failed to produce putative transformants. Also no putative transgenics of barley Scralett, wheat Torka and triticale Wanad were obtained after transformation with EHA101 and selection on higromycine. Selection with kanamycin and hygromycin + kanamycin after transformation with EHA101 and LBA4404 respectively also failed to give positive results.
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