Effects of knocking out three anthocyanin modification genes on the blue pigmentation of gentian flowers

Abstract Genome editing by the CRISPR/Cas9 system has recently been used to produce gene knockout lines in many plant species. We applied this system to analyze Japanese gentian plants that produce blue flowers because of the accumulation of a polyacylated anthocyanin, gentiodelphin. Mutant lines in...

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Autores principales:	Keisuke Tasaki, Atsumi Higuchi, Aiko Watanabe, Nobuhiro Sasaki, Masahiro Nishihara
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2019
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/60e39567db9f4477a354474357ab62b7
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spelling	oai:doaj.org-article:60e39567db9f4477a354474357ab62b72021-12-02T15:08:20ZEffects of knocking out three anthocyanin modification genes on the blue pigmentation of gentian flowers10.1038/s41598-019-51808-32045-2322https://doaj.org/article/60e39567db9f4477a354474357ab62b72019-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-019-51808-3https://doaj.org/toc/2045-2322Abstract Genome editing by the CRISPR/Cas9 system has recently been used to produce gene knockout lines in many plant species. We applied this system to analyze Japanese gentian plants that produce blue flowers because of the accumulation of a polyacylated anthocyanin, gentiodelphin. Mutant lines in which anthocyanin modification genes were knocked out were examined to assess the contribution of each gene to the blue pigmentation of flowers. The targeted genes encoded anthocyanin 5-O-glycosyltransferase (Gt5GT), anthocyanin 3′-O-glycosyltransferase (Gt3′GT), and anthocyanin 5/3′-aromatic acyltransferase (Gt5/3′AT). The Gt5GT knockout lines accumulated delphinidin 3G, whereas the Gt3′GT knockout lines accumulated delphinidin 3G-5CafG as the major flower pigment. Knocking out Gt5/3′AT resulted in the accumulation of delphinidin 3G-5G-3′G and delphinidin 3G-5G as the primary and secondary pigments, respectively. These results indicated the existence of two pathways mediating the modification of delphinidin 3G-5G in flowers, with one involving a glycosylation by 3′GT and the other involving an acylation by 5/3′AT. The Gt5GT, Gt3′GT, and Gt5/3′AT transformants produced pale red violet, dull pink, and pale mauve flowers, respectively, unlike the vivid blue flowers of wild-type plants. Thus, the glycosylation and subsequent acylation of the 3′-hydroxy group of the B-ring in delphinidin aglycone is essential for the development of blue gentian flowers.Keisuke TasakiAtsumi HiguchiAiko WatanabeNobuhiro SasakiMasahiro NishiharaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 9, Iss 1, Pp 1-10 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Keisuke Tasaki Atsumi Higuchi Aiko Watanabe Nobuhiro Sasaki Masahiro Nishihara Effects of knocking out three anthocyanin modification genes on the blue pigmentation of gentian flowers
description	Abstract Genome editing by the CRISPR/Cas9 system has recently been used to produce gene knockout lines in many plant species. We applied this system to analyze Japanese gentian plants that produce blue flowers because of the accumulation of a polyacylated anthocyanin, gentiodelphin. Mutant lines in which anthocyanin modification genes were knocked out were examined to assess the contribution of each gene to the blue pigmentation of flowers. The targeted genes encoded anthocyanin 5-O-glycosyltransferase (Gt5GT), anthocyanin 3′-O-glycosyltransferase (Gt3′GT), and anthocyanin 5/3′-aromatic acyltransferase (Gt5/3′AT). The Gt5GT knockout lines accumulated delphinidin 3G, whereas the Gt3′GT knockout lines accumulated delphinidin 3G-5CafG as the major flower pigment. Knocking out Gt5/3′AT resulted in the accumulation of delphinidin 3G-5G-3′G and delphinidin 3G-5G as the primary and secondary pigments, respectively. These results indicated the existence of two pathways mediating the modification of delphinidin 3G-5G in flowers, with one involving a glycosylation by 3′GT and the other involving an acylation by 5/3′AT. The Gt5GT, Gt3′GT, and Gt5/3′AT transformants produced pale red violet, dull pink, and pale mauve flowers, respectively, unlike the vivid blue flowers of wild-type plants. Thus, the glycosylation and subsequent acylation of the 3′-hydroxy group of the B-ring in delphinidin aglycone is essential for the development of blue gentian flowers.
format	article
author	Keisuke Tasaki Atsumi Higuchi Aiko Watanabe Nobuhiro Sasaki Masahiro Nishihara
author_facet	Keisuke Tasaki Atsumi Higuchi Aiko Watanabe Nobuhiro Sasaki Masahiro Nishihara
author_sort	Keisuke Tasaki
title	Effects of knocking out three anthocyanin modification genes on the blue pigmentation of gentian flowers
title_short	Effects of knocking out three anthocyanin modification genes on the blue pigmentation of gentian flowers
title_full	Effects of knocking out three anthocyanin modification genes on the blue pigmentation of gentian flowers
title_fullStr	Effects of knocking out three anthocyanin modification genes on the blue pigmentation of gentian flowers
title_full_unstemmed	Effects of knocking out three anthocyanin modification genes on the blue pigmentation of gentian flowers
title_sort	effects of knocking out three anthocyanin modification genes on the blue pigmentation of gentian flowers
publisher	Nature Portfolio
publishDate	2019
url	https://doaj.org/article/60e39567db9f4477a354474357ab62b7
work_keys_str_mv	AT keisuketasaki effectsofknockingoutthreeanthocyaninmodificationgenesonthebluepigmentationofgentianflowers AT atsumihiguchi effectsofknockingoutthreeanthocyaninmodificationgenesonthebluepigmentationofgentianflowers AT aikowatanabe effectsofknockingoutthreeanthocyaninmodificationgenesonthebluepigmentationofgentianflowers AT nobuhirosasaki effectsofknockingoutthreeanthocyaninmodificationgenesonthebluepigmentationofgentianflowers AT masahironishihara effectsofknockingoutthreeanthocyaninmodificationgenesonthebluepigmentationofgentianflowers
_version_	1718388187777204224

Effects of knocking out three anthocyanin modification genes on the blue pigmentation of gentian flowers

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