Anthocyanin Fruit encodes an R2R3-MYB transcription factor, SlAN2-like, activating the transcription of SlMYBATV to fine-tune anthocyanin content in tomato fruit
Anthocyanin Fruit encodes an R2R3-MYB transcription factor, SlAN2-like, activating the transcription of SlMYBATV to fine-tune anthocyanin content in tomato fruit
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Anthocyanin Fruit 编码一种类似 SlAN2 的 R2R3-MYB 转录因子，激活 SlMYBATV 的转录来微调番茄果实中的花青素含量
DOI：
10.1111/nph.16272
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发表时间：
2019-11-21
期刊：
影响因子：
9.4
通讯作者：
Qiu, Zhengkun
中科院分区：
文献类型：
--
作者：
Yan, Shuangshuang;Chen, Na;Qiu, Zhengkun
野生番茄具有花色苷果实（Aft）和紫黑色果实（atv）的特征，可提高果实花色苷含量。然而，Aft基因座的基因和Aft和atv的作用机制在很大程度上仍然未知。在这项研究中，Aft基因座被精细定位到10号染色体上约145 kb的区间，不包括先前被建议作为候选基因的SlAN2（Solyc10g086250）、SlANT1（Solyc10g086260）和SlANT1-like（Solyc10g086270）。因此，R2R3-MYB转录因子SlAN 2样（Solyc10g086290）被认为是Aft的最佳候选基因。与野生型番茄相比，CRISPR/Cas9介导的SlAN 2样突变体显示出与多个花青素相关基因的下调相关的花青素的低得多的积累，表明SlAN 2样是Aft表型的原因。SlMYBATV的抑制功能也通过CRISPR/Cas9方法得到证实。酵母双杂交测定揭示了SlMYBATV与bHLH蛋白SlJAF13相互作用。此外，酵母单杂交和双荧光素酶瞬时表达测定表明，Aft直接结合到SlMYBATV启动子并激活其表达。本文的结果提供了提高番茄果实中花色素苷含量的候选基因。这项研究还提供了深入了解涉及Aft-SlMYBATV途径的机制，该途径可以微调番茄果实中花青素苷的积累。
Anthocyanin fruit (Aft) and atroviolacea (atv) were characterized in wild tomato and can enhance anthocyanin content in tomato fruit. However, the gene underlying the Aft locus and the mechanism by which Aft and atv act remain largely unknown. In this study, the Aft locus was fine-mapped to an approximately 145-kb interval on chromosome 10, excluding SlAN2 (Solyc10g086250), SlANT1 (Solyc10g086260) and SlANT1-like (Solyc10g086270), which have previously been suggested as candidates. Thus, the R2R3-MYB transcription factor SlAN2-like (Solyc10g086290) was considered the best candidate gene for Aft. The CRISPR/Cas9-mediated SlAN2-like mutants show a much lower accumulation of anthocyanins associated with the downregulation of multiple anthocyanin-related genes compared to the wild-type tomato, indicating that SlAN2-like is responsible for the Aft phenotype. The repressive function of SlMYBATV also was confirmed through the CRISPR/Cas9 approach. A yeast-two-hybrid assay revealed that SlMYBATV interacts with the bHLH protein SlJAF13. Furthermore, yeast-one-hybrid and dual-luciferase transient expression assays showed that Aft directly binds to the SlMYBATV promoter and activates its expression. The results herein provide candidate genes to enhance anthocyanin content in tomato fruit. This research also provides insight into a mechanism involving the Aft-SlMYBATV pathway that fine-tunes anthocyanin accumulation in tomato fruit.