A single-nucleotide polymorphism in WRKY33 promoter is associated with the cold sensitivity in cultivated tomato
A single-nucleotide polymorphism in WRKY33 promoter is associated with the cold sensitivity in cultivated tomato
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WRKY33 启动子中的单核苷酸多态性与栽培番茄的冷敏感性相关
DOI：
10.1111/nph.18403
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发表时间：
2022-08-13
期刊：
影响因子：
9.4
通讯作者：
Zhou, Jie
中科院分区：
文献类型：
--
作者：
Guo, Mingyue;Yang, Fengjun;Zhou, Jie
顺式调控区的自然变异通常通过改变基因表达来影响作物表型。然而，启动子突变如何影响基因表达和作物抗逆性的机制仍然知之甚少。本研究通过对栽培番茄及其野生近缘种的RNA测序（RNA-Seq）数据分析和逆转录实时荧光定量PCR验证，发现WRKY 33基因在冷敏感栽培番茄Solanum lycopersicum L.“Ailsa克雷格”，但显着诱导耐寒野生番茄的亲戚Solanum habrochaites LA 1777和Solanum pennellii LA 0716在低温胁迫下。SlWRKY 33或ShWRKY 33的过表达正调节番茄的耐冷性。SlWRKY 33启动子中的关键W-box的变异导致SlWRKY 33在冷胁迫下失去自转录功能。整合RNA-Seq和染色质免疫沉淀测序数据的分析表明，SlWRKY 33直接靶向并诱导多种激酶、转录因子和分子伴侣基因，例如CDPK 11、MYBS 3和BAG 6，从而增强耐寒性。此外，热和灰霉病诱导的WRKY 33在野生和栽培番茄的表达是独立的关键W盒的变化。我们的研究结果表明，核苷酸多态性的顺式调控区是至关重要的栽培番茄和野生番茄植物之间的不同冷敏感性。
Natural variations in cis-regulatory regions often affect crop phenotypes by altering gene expression. However, the mechanism of how promoter mutations affect gene expression and crop stress tolerance is still poorly understood. In this study, by analyzing RNA-sequencing (RNA-Seq) data and reverse transcription quantitative real-time PCR validation in the cultivated tomato and its wild relatives, we reveal that the transcripts of WRKY33 are almost unchanged in cold-sensitive cultivated tomato Solanum lycopersicum L. 'Ailsa Craig' but are significantly induced in cold-tolerant wild tomato relatives Solanum habrochaites LA1777 and Solanum pennellii LA0716 under cold stress. Overexpression of SlWRKY33 or ShWRKY33 positively regulates cold tolerance in tomato. Variant of the critical W-box in SlWRKY33 promoter results in the loss of self-transcription function of SlWRKY33 under cold stress. Analysis integrating RNA-Seq and chromatin immunoprecipitation sequencing data reveals that SlWRKY33 directly targets and induces multiple kinases, transcription factors, and molecular chaperone genes, such as CDPK11, MYBS3, and BAG6, thus enhancing cold tolerance. In addition, heat- and Botrytis-induced WRKY33s expression in both wild and cultivated tomatoes are independent of the critical W-box variation. Our findings suggest nucleotide polymorphism in cis-regulatory regions is crucial for different cold sensitivity between cultivated and wild tomato plants.