Transient Expression of Anthocyanin in Developing Wheat Coleoptile by Maize C1 and B-peru Regulatory Genes for Anthocyanin Synthesis.

Abstract

Maize transcription factors for the genes of anthocyanin synthesis pathway, C1/B-peru, were delivered into developing wheat coleoptiles by a particle bombardment method. Anthocyanin pigments were induced as discrete red spots and their number reached about 60 spots per coleoptile, compared to about 20 diffused blue GUS spots, which were induced by the GusA gene transferred concomitantly. Coleoptiles of seedlings collected 36 h after germination were most suitable tissue for the expression of delivered foreign genes. One to five µg of plasmid DNA for coating gold particles was sufficient for induction of anthocyanin and GUS, indicating that the transferred gene was expressed efficiently in coleoptile cells. Helium gas pressure (900, 1100, 1300 or 1500 pounds per square inch) and distances (10, 15, or 20 cm) between the stopping screen and coleoptile did not affect significantly the efficiency of gene transfer. Seedlings arranged in a circle with a 1-cm radius on a MS medium plate were targeted well by gold particles. The results showed that the wheat coleoptile was a good target tissue for transient assay of wheat genes and that C1/B-peru can be used as a reporter gene.

TaMYB3, encoding a functional MYB transcriptor, isolated from the purple pericarp of Triticum aestivum

“…TaMYB3 was very similar to ZmC1 in the evolutionary analysis, and in previous research, ZmC1 was shown to induce anthocyanin biosynthesis when coexpressed with the bHLH gene (Cone et al 1986;Ahmed et al 2003). In the present study, the bHLH regulators ZmR and ZmC1 were isolated from maize to compare the functions of TaMYB3 and ZmC1.…”

“…The sequence-conservation-based strategy used to isolate the homologous gene was very efficient. The transient expression of TaMYB3 induced anthocyanin biosynthesis in the pericarp of cultivar Gy115 during dark treatment, with the co-operation of the bHLH transcription factor ZmR, which is similar to ZmC1 (Ahmed et al 2003). TaMYB3 is expressed in purple grains and is induced by light.…”

“…To transform the cells, the plasmids were shot into them with a gene gun, according to the method of Ahmed, for functional verification (Ahmed et al 2003). The grains were selected for bombardment 14 days after blossoming.…”

Identification of a candidate gene for Rc-D1, a locus controlling red coleoptile colour in wheat

“…In previous research, ZmC1 has been shown to induce anthocyanin biosynthesis with the coexpression of the bHLH gene ZmR (Ahmed et al 2003;. In the present experiment, the bHLH regulators ZmR and ZmC1 were isolated from maize to compare the function of TaMYB-D1 with that of ZmC1.…”

“…The transient plasmids pBract214-TaMYB-D1, pBract214-ZmR (bHLH gene in maize), and pBract214-ZmC1 (MYB gene in maize) were constructed using the Gateway Cloning Kit (Invitrogen, USA). The plasmids were delivered into the coleoptiles of 'Opata' by particle bombardment according to the methods of Ahmed et al (2003). Four plasmids were delivered: pBract214-ZmR, pBract214-ZmC1, pBract214-TaMYB-D1, and pBract-control.…”