title: De Novo Transcriptome Analysis to Identify Anthocyanin Biosynthesis Genes Responsible for Tissue-Specific Pigmentation in Zoysiagrass (Zoysia japonica Steud.).
abstract: Zoysiagrass (Zoysia japonica Steud.) is commonly found in temperate climate regions and widely used for lawns, in part, owing to its uniform green color. However, some zoysiagrass cultivars accumulate red to purple pigments in their spike and stolon tissues, thereby decreasing the aesthetic value. Here we analyzed the anthocyanin contents of two zoysiagrass cultivars 'Anyang-jungji' (AJ) and 'Greenzoa' (GZ) that produce spikes and stolons with purple and green colors, respectively, and revealed that cyanidin and petunidin were primarily accumulated in the pigmented tissues. In parallel, we performed a de novo transcriptome assembly and identified differentially expressed genes between the two cultivars. We found that two anthocyanin biosynthesis genes encoding anthocyanidin synthase (ANS) and dihydroflavonol 4-reductase (DFR) were preferentially upregulated in the purple AJ spike upon pigmentation. Both ANS and DFR genes were also highly expressed in other zoysiagrass cultivars with purple spikes and stolons, but their expression levels were significantly low in the cultivars with green tissues. We observed that recombinant ZjDFR1 and ZjANS1 proteins successfully catalyze the conversions of dihydroflavonols into leucoanthocyanidins and leucoanthocyanidins into anthocyanidins, respectively. These findings strongly suggest that upregulation of ANS and DFR is responsible for tissue-specific anthocyanin biosynthesis and differential pigmentation in zoysiagrass. The present study also demonstrates the feasibility of a de novo transcriptome analysis to identify the key genes associated with specific traits, even in the absence of reference genome information.
authors.affiliation: Department of Plant Science, Seoul National University, Seoul, 151-921, Korea.
authors.forename: Jong Hwa
authors.initials: JH
authors.lastname: Ahn
authors.affiliation: Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, 151-921, Korea.
authors.forename: June-Sik
authors.initials: JS
authors.lastname: Kim
authors.affiliation: Interdisciplinary Program in Agricultural Genomics, Seoul National University, Seoul, 151-921, Korea.
authors.forename: Seungill
authors.initials: S
authors.lastname: Kim
authors.affiliation: Interdisciplinary Program in Agricultural Genomics, Seoul National University, Seoul, 151-921, Korea.
authors.forename: Hye Yeon
authors.initials: HY
authors.lastname: Soh
authors.affiliation: Department of Plant Science, Seoul National University, Seoul, 151-921, Korea.
authors.forename: Hosub
authors.initials: H
authors.lastname: Shin
authors.affiliation: Department of Plant Science, Seoul National University, Seoul, 151-921, Korea.
authors.forename: Hosung
authors.initials: H
authors.lastname: Jang
authors.affiliation: Department of Plant Science, Seoul National University, Seoul, 151-921, Korea.
authors.forename: Ju Hyun
authors.initials: JH
authors.lastname: Ryu
authors.affiliation: FnP Co., Ltd, Jeungpyeong, 368-811, Korea.
authors.forename: Ahyeong
authors.initials: A
authors.lastname: Kim
authors.affiliation: FnP Co., Ltd, Jeungpyeong, 368-811, Korea.
authors.forename: Kil-Young
authors.initials: KY
authors.lastname: Yun
authors.affiliation: FnP Co., Ltd, Jeungpyeong, 368-811, Korea.
authors.forename: Shinje
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Plant Science, Seoul National University, Seoul, 151-921, Korea; Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, 151-921, Korea.
authors.forename: Ki Sun
authors.initials: KS
authors.lastname: Kim
authors.affiliation: Department of Plant Science, Seoul National University, Seoul, 151-921, Korea; Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, 151-921, Korea; Interdisciplinary Program in Agricultural Genomics, Seoul National University, Seoul, 151-921, Korea.
authors.forename: Doil
authors.initials: D
authors.lastname: Choi
authors.affiliation: Department of Plant Science, Seoul National University, Seoul, 151-921, Korea; Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, 151-921, Korea; Interdisciplinary Program in Agricultural Genomics, Seoul National University, Seoul, 151-921, Korea.
authors.forename: Jin Hoe
authors.initials: JH
authors.lastname: Huh
mesh_terms: D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000378:metabolism; D000872:Anthocyanins / Q000032:analysis / Q000096:biosynthesis*; D055162:Biocatalysis; D002851:Chromatography, High Pressure Liquid; D020869:Gene Expression Profiling; D017343:Genes, Plant; D013058:Mass Spectrometry; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D010858:Pigmentation / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D006109:Poaceae / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D015854:Up-Regulation
chemical_list: D000872:Anthocyanins; D010940:Plant Proteins; D011994:Recombinant Proteins; C017154:cyanidin; D000429:Alcohol Oxidoreductases; C056570:dihydroflavanol 4-reductase; D010105:Oxygenases; C118159:anthocyanidin synthase; C473206:petunidin