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ACTA HORTICULTURAE SINICA ›› 2013, Vol. 40 ›› Issue (12): 2419-2428.

• Vegetables • Previous Articles     Next Articles

Cloning of Arabidopsis At-pri-miR828 Gene and Its Genetic Transformation into Tomato

 JIA  Xiao-Yun, YU  Zhi-Qin, LIANG  Jian-Ping, TANG  Gui-Liang, JIN  Lei-Hao, ZHANG   Li, HE  Li-Heng, LI  Run-Zhi   

  1. (1College of Life Sciences,Shanxi Agricultural University,Taigu,Shanxi 030801,China;2Department of Biological
    Sciences,Michigan Technological University,Houghton,MI 49931,USA;3Henan Agricultural University,Zhengzhou
    450002,China;4College of Agriculture,Shanxi Agricultural University,Taigu,Shanxi 030801,China)
  • Online:2013-12-25 Published:2013-12-25

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

CLC Number: