关键词: 番茄, miR828, Sly-myb-like1, 花青素, 实时定量PCR

Abstract: MicroRNA828（miR828）is a newly identified small RNA whose biological functions are
still not very clear. To explore the biological functions of miR828，particularly in tomato，the At-primiR828
gene was isolated from Arabidopsis thaliana，and cloned into the corresponding site of the
pCAMBIA2300 to construct the plant expression vector pC2300-pOT2-At-pri-miR828. This At-pri-miR828 overexpressing vector was then introduced into tomato（Solanum lycopersicum Mill.‘Ailsa Graig’）using
Agrobacterium-mediated transformation. Total of 9 transgenic lines and 67 transgenic plants carrying the
At-pri-miR828 gene under the control of the 35S promoter were generated. PCR amplification showed that
At-pri-miR828 was integrated into the genome DNA of transgenic tomatoes. Real-time PCR analysis
demonstrated that miR828 mRNA level in the transgenic tomatoes was increased greatly. Accordingly，
expression of Sly-myb-like1，a potential target gene of miR828 predicted by informatics，was suppressed.
Further anthocyanin content analysis revealed that anthocyanin content was greatly reduced in miR828
overexpressed transgenic plants，demonstrating miR828 might function in the anthocyanin biosynthesis of
tomato.

Key words: tomato, miR828, Sly-myb-like1, anthocyanin, real-time PCR