Study on transformation of tobacco plant with flower symmetry gene LjCYC3 in Lotus japonicus

Abstract

According to LjCYC3 gene controlling flower symmetry in Lotus japonicus, the antisense gene expression vector was constructed, and by using Agrobacterium-mediated transformation of tabacoo with justice-antisense LjCYC3 gene, flower patterns changed in transgenic tobacco plants were obtained, and then three transgenic plants and a wild-type plant were selected for Q-PCR analysis according to phenotype. PCR analysis showed that transformation rates of justice gene and antisense gene were 62.5% and 71% in transgenic plants, respectively. Phenotypic observations showed that 30% of justice transgenic plants mainly appeared to a decrease number in petals and stamens, however, 40% of the antisense transgenic plants appeared to have two flowers fusion to some degree, and color fading for petals and sepals, the head of stamens having petal-shaped structures, and two pistils being connate with stamens. The Q-PCR analysis results showed that foreign genes were highly expressed in the justice transgenic plants, and the more expression increased, the more pronounced pattern changed. The foreign gene was also expressed in the antisense transgenic plants, and as the reduction of the expression, pattern change was even more evident. Compared with the justice gene expression quantity, the antisense gene expression quantity was 150 times lower. The results above lay a good foundation for transformation of flowering plants with the LjCYC3 gene to change flower pattern and improve flower varieties for the next stage.

Keywords

• <italic>Lotus japonicus</italic>
• <italic>LjCYC3</italic> gene
• justice-antisense expression vector
• tobacco