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Two of these genes, ASR<\/jats:styled-content>1<\/jats:italic> and ASR<\/jats:styled-content>2,<\/jats:italic> are inactivated by two different single base mutations in their coding sequence. All three of these genes are absent in the white flowering species P.\u00a0axillaris N<\/jats:italic> and P.\u00a0parodii<\/jats:italic>, in the red flowering P.\u00a0exserta<\/jats:italic>, and in several Petunia hybrida<\/jats:italic> lines, including R27 and W115. P.\u00a0violacea<\/jats:italic> and other P<\/jats:italic>.\u00a0hybrida<\/jats:italic> lines (M1, V30, and W59) instead harbor functional copies of the ASR<\/jats:styled-content><\/jats:italic> genes. Comparative, functional and phylogenic analysis of anthocyanin R2R3\u2010MYB<\/jats:styled-content> genes strongly suggest that the ASR<\/jats:styled-content><\/jats:italic> genes cluster is a duplication of the genomic fragment containing the other three R2R3\u2010MYB<\/jats:styled-content> genes with roles in pigmentation that were previously defined, the ANTHOCYANIN<\/jats:styled-content>4\u2010DEEP PURPLE<\/jats:styled-content><\/jats:italic>\u2010PURPLE HAZE<\/jats:styled-content><\/jats:italic> (AN<\/jats:styled-content>4<\/jats:italic>\u2010DPL<\/jats:styled-content><\/jats:italic>\u2010PHZ<\/jats:styled-content><\/jats:italic>) cluster. An investigation of the genomic fragments containing anthocyanin MYB<\/jats:styled-content>s in different Petuni<\/jats:italic>a accessions reveals that massive rearrangements have taken place, resulting in large differences in the regions surrounding these genes, even in closely related species. Yeast two\u2010hybrid assays showed that the ASR<\/jats:styled-content> proteins can participate in the WMBW<\/jats:styled-content> (WRKY<\/jats:styled-content>, MYB<\/jats:styled-content>, B\u2010HLH<\/jats:styled-content>, and WDR<\/jats:styled-content>) anthocyanin regulatory complex by interacting with the transcription factors AN<\/jats:styled-content>1 and AN<\/jats:styled-content>11. All three ASR<\/jats:styled-content>s can induce anthocyanin synthesis when ectopically expressed in P.\u00a0hybrida<\/jats:italic> lines, but ASR<\/jats:styled-content>1 appeared to be the most effective. The expression patterns of ASR<\/jats:styled-content>1<\/jats:italic> and ASR<\/jats:styled-content>2<\/jats:italic> cover several different petunia tissues with higher expression at early stages of bud development. In contrast, ASR<\/jats:styled-content>3<\/jats:italic> is only weakly expressed in the stigma, ovary, and anther filaments. The characterization of these novel ASR<\/jats:styled-content><\/jats:italic> MYB<\/jats:styled-content> genes completes the picture of the MYB<\/jats:styled-content> members of the petunia anthocyanin regulatory MBW<\/jats:styled-content> complex and suggests possible mechanisms of the diversification of pigmentation patterns during plant evolution.<\/jats:p>","DOI":"10.1002\/pld3.114","type":"journal-article","created":{"date-parts":[[2019,1,21]],"date-time":"2019-01-21T13:52:57Z","timestamp":1548078777000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":41,"title":["Identification and functional analysis of three new anthocyanin R2R3\u2010MYB<\/scp> genes in Petunia<\/i>"],"prefix":"10.1002","volume":"3","author":[{"given":"Hechen","family":"Zhang","sequence":"first","affiliation":[{"name":"Horticulture Research Institute Henan Academy of Agricultural Sciences Zhengzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ronald","family":"Koes","sequence":"additional","affiliation":[{"name":"Department of Plant Development and (Epi) Genetics Swammerdam Institute for Life Sciences University of Amsterdam Amsterdam The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongquan","family":"Shang","sequence":"additional","affiliation":[{"name":"Horticulture Research Institute Henan Academy of Agricultural Sciences Zhengzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhenzhu","family":"Fu","sequence":"additional","affiliation":[{"name":"Horticulture Research Institute Henan Academy of Agricultural Sciences Zhengzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Limin","family":"Wang","sequence":"additional","affiliation":[{"name":"Horticulture Research Institute Henan Academy of Agricultural Sciences Zhengzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyu","family":"Dong","sequence":"additional","affiliation":[{"name":"Horticulture Research Institute Henan Academy of Agricultural Sciences Zhengzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Zhang","sequence":"additional","affiliation":[{"name":"Horticulture Research Institute Henan Academy of Agricultural Sciences Zhengzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valentina","family":"Passeri","sequence":"additional","affiliation":[{"name":"Department of Plant Development and (Epi) Genetics Swammerdam Institute for Life Sciences University of Amsterdam Amsterdam The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanbang","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Plant Science School of Agriculture and Biology Shanghai Jiao Tong University Shanghai 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