Scientia Agricultura Sinica ›› 2015, Vol. 48 ›› Issue (7): 1392-1404.doi: 10.3864/j.issn.0578-1752.2015.07.14

• HORTICULTURE • Previous Articles     Next Articles

Cloning and Expression Analyses of R2R3-MYB Genes Related to Anthocyanin Biosynthesis in Rose

ZHAO Jia, LIU Rong, YANG Fan, LI Xin, LIU Hou-sheng, YAN Qian, XIAO Yue-hua   

  1. Biotechnology Research Center, Southwest University/Chongqing Key Laboratory of Application and Safety Control of Genetically Modified Crops, Chongqing 400716
  • Received:2014-08-26 Online:2015-04-01 Published:2015-04-01

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Abstract: 【Objective】Anthocyanin is a major factor conferring pink or red to rose flower. R2R3-MYB proteins are key transcription factors controlling anthocyanin biosynthesis. This work aimed to clone homologous genes of anthocyanin-related R2R3-MYB protein from rose flowers, and further to analyze their relatedness to anthocyanin biosynthesis and coloration in rose petals, which may lay a foundation for gene engineering improvement of flower.【Method】With a degenerate primer designed according to the conserved sequence of anthocyanin-related R2R3-MYB proteins from various plants, complete coding sequences of corresponding homologous genes were amplified and cloned by using 3′-RACE and Y-RACE methods. Multiple sequence alignment and phylogenetic analysis were performed using typical sequences from the 4th and 6th subfamily of plant R2R3-MYB proteins (Sg4 and Sg6) and Arabidopsis R2R3-MYB proteins related to secondary metabolism with deduced proteins of the cloned genes. To elucidate the relatedness of the cloned genes to anthocyanin biosynthesis and coloration in rose petals, anthocyanin contents and expression levels of the two cloned R2R3-MYB protein genes were determined in rose petals of different colors. 【Result】Two R2R3-MYB protein genes (RhMYBs4-1 and RhMYBs6-1, GenBank accession Nos, KJ664810 and KJ664811, respectively) were cloned from red petals of rose ‘Red Success’. Sequence analyses indicated that RhMYBs4-1 and RhMYBs6-1, both conserved in the R2R3-MYB domain, were homologous to Sg4 and Sg6 R2R3-MYB proteins, respectively. RhMYBs4-1 contained the C1, C2 repressors and zinc finger domains typical of Sg4 R2R3-MYB proteins, whereas RhMYBs6-1 had the signature motifs ((A/S/G)NDV and KPRPR(T/S)) of Sg6 R2R3-MYB subfamily. Expression analyses showed that both RhMYBs4-1 and RhMYBs6-1 were expressed at a high level in the red petals of rose ‘Red Success’, but at low levels in leaves and stamens. Among rose petals of different colors, red petals had the highest anthocyanin contents, and the highest expression levels of both RhMYBs4-1 and RhMYBs6-1 genes. In pink rose petals, the anthocyanin content was less than 10% of that in red petals and the RhMYBs4-1 expression level was very low, whereas the expression level of RhMYBs6-1 in pink petals was comparable to that in red petals. 【Conclusion】Results of this study indicated that the RhMYBs4-1 and RhMYBs6-1 genes encoded Sg4 and Sg6 R2R3-MYB proteins, respectively. Both RhMYBs4-1 and RhMYBs6-1 were highly expressed in red petals and might be important regulators of anthocyanin biosynthesis and coloration in rose petals.

Key words: Rosa hybrida, petal color, anthocyanin, R2R3-MYB protein, gene expression

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