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GrainGenes Reference Report: TAG-105-201

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Reference	TAG-105-201
Title	Competence of oat (Avena sativa L.) shoot apical meristems for integrative transformation, inherited expression, and osmotic tolerance of transgenic lines containing hva1
Journal	Theoretical and Applied Genetics
Year	2002
Volume	105
Pages	201-208
Author	Maqbool SB [ Show all 8 ]
Abstract	Summary: Three oat (Avena sativa L.) cultivars have been successfully transformed using an efficient and reproducible in vitro culture system for differentiation of multiple shoots from shoot apical meristems. The transformation was performed using microprojectile bombardment with two plasmids (pBY520 and pAct1-D) containing linked (hva1-bar) and non-linked (gus) genes. The hva1 and bar genes cointegrated with a frequency of 100% as expected, and 61.6% of the transgenic plants carried all three genes. Molecular and biochemical analyses in R0, R1 and R2 progenies confirmed stable integration and expression of all transgenes. Localization of the GUS protein in R0 and R1 plants revealed that high-expression of gus occurred in vascular tissues and in the pollen grains of mature flowers. The constitutive expression of HVA1 protein was observed at all developmental stages of transgenic plants, and was particularly stronger during the early seedling stages. R2 progeny of five independent transgenic lines was tested in vitro for tolerance to osmotic (salt and mannitol) stresses. As compared to non-transgenic control plants, transgenic plants maintained a higher growth and showed significantly (P < 0.05) increased tolerance to stress conditions. Less than 10% of transgenic plants showed symptoms of wilting or death of leaves and, when these symptoms present were delayed in transgenic plants as compared to 80% of non-transgenic plants, either wilted or died. These symptoms confirmed the increased in vitro tolerance in hva1-expressing transgenic plants to non-transgenic plants, providing strong evidence that the HVA1 protein may play an important role in the protection of oats against salinity and possible water-deficiency stress conditions
External Databases	http://link.springer.de/link/service/journals/00122/contents/02/00984/
Keyword	[ Hide all but 1 of 57 ] apical meristem avena avena sativa bar bar gene barley bombardment constitutive expression death developmental stage developmental stages differentiation efficient flower frequencies gene grains growth gus hva1 in vitro in vitro culture in-vitro morphogenesis integration integrative transformation localization mannitol microprojectile bombardment multiple multiple shoots oat oat (avena sativa l.) oats plasmid pollen progeny protection protein r2 salinity salt salt and mannitol-stress seedling stage shoot apical meristem stable integration stress stress conditions symptoms tissue tolerance transformation transgene transgenic vascular tissue vitro water-deficit wilting

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