Premium
Endosperm‐specific expression of green fluorescent protein driven by the hordein promoter is stably inherited in transgenic barley ( Hordeum vulgare ) plants
Author(s) -
Cho MyeongJe,
Choi HaeWoon,
Jiang Wen,
Ha Chi D.,
Lemaux Peggy G.
Publication year - 2002
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1034/j.1399-3054.2002.1150117.x
Subject(s) - hordein , green fluorescent protein , endosperm , biology , hordeum vulgare , transgene , microbiology and biotechnology , callus , gene , gene expression , genetically modified rice , genetically modified crops , genetics , botany , poaceae
The expression of green fluorescent protein (GFP) and its inheritance were studied in transgenic barley ( Hordeum vulgare L.) plants transformed with a synthetic green fluorescent protein gene [ sgfp (S65T)] driven by either a rice actin promoter or a barley endosperm‐specific d ‐hordein promoter. The gene encoding phosphinothricin acetyltransferase ( bar ), driven by the maize ubiquitin promoter and intron, was used as a selectable marker to identify transgenic tissues. Strong GFP expression driven by the rice actin promoter was observed in callus cells and in a variety of tissues of T 0 plants transformed with the sgfp (S65T)‐containing construct. GFP expression, driven by the rice actin promoter, was observed in 14 out of 17 independent regenerable transgenic callus lines; however, expression was gradually lost in T 0 and later generation progeny of diploid lines. Stable GFP expression was observed in T 2 progeny from only 6 out of the 14 (43%) independent GFP‐expressing callus lines. Four of the 8 lines not expressing GFP in T 2 progeny, lost GFP expression during T 0 plant regeneration from calli; one lost GFP expression in the transition from the T 0 to T 1 generations and three lines were sterile. Similarly, expression of bar driven by the maize ubiquitin promoter was lost in T 1 progeny; only 21 out of 26 (81%) independent lines were Basta‐resistant. In contrast to actin‐driven expression, GFP expression driven by the d ‐hordein promoter exhibited endosperm‐specificity. All seven lines transformed with d ‐hordein‐driven GFP (100%) expressed GFP in the T 1 and T 2 generations, regardless of ploidy levels, and expression segregated in a Mendelian fashion. We conclude that the sgfp (S65T) gene was successfully transformed into barley and that GFP expression driven by the d ‐hordein promoter was more stable in its inheritance pattern in T 1 and T 2 progeny than that driven by the rice actin promoter or the bar gene driven by the maize ubiquitin promoter.