CmNLP7 interacts with CmPP6 to suppress flowering time in Chrysanthemum

IF 5.7 1区 农林科学 Q1 HORTICULTURE Horticultural Plant Journal Pub Date : 2024-09-14 DOI:10.1016/j.hpj.2023.09.012
Guohui Wang, Juanjuan Wang, Mengru Yin, Yu Zhang, Jiaxin Zhang, Chaona Si, Sumei Chen, Zhiyong Guan, Shuang Zhao, Fei Zhang, Weiming Fang, Fadi Chen, Jiafu Jiang
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Abstract

Nitrogen (N) is a key component in plants and their biological macromolecules, having a profound effect on developmental stages, such as germination, vegetative growth, and flowering. However, the mechanism of nitrogen-regulated flowering time remains unclear. In this study, CmNLP7 was isolated from the chrysanthemum cultivar ‘Jinba’ and was characterized. CmNLP7 is a transcription factor localized in the nucleus but has no transcriptional activity. Tissue expression pattern analysis showed that CmNLP7 was mainly transcribed in leaves and roots. Knocking down CmNLP7 through the artificial-miRNA method in chrysanthemum resulted in early flowering under optimal nitrogen (ON) and low nitrogen (LN) conditions; whereas overexpression lines showed delayed flowering under LN conditions. Transcriptome sequencing analysis showed that the nitrate transporters NRT2.5, NPF3.1, and NPF4.6; SBP-like genes SPL7 and SPL12, and flowering integration factor FT were significantly up-regulated in the knockdown lines. Based on the KEGG pathway enrichment analysis, the differentially transcribed genes were enriched in phenylpropanoid biosynthesis and starch and sucrose metabolism pathways, which indicated their alleged function in nitrogen-regulated flowering and development in chrysanthemum. Furthermore CmPP6 as a homolog of the Arabidopsis phosphatase PP6, was verified as an interacting protein of CmNLP7 by yeast two-hybrid, BiFC, pull-down and Biacore in vitro and in vivo, and the knockdown line of CmPP6 (amiR-CmPP6) flowered earlier compared to that of the wild-type chrysanthemum ‘Jinba’. Collectively, these results demonstrated that CmPP6 interacts with CmNLP7 to regulate chrysanthemum flowering, and CmNLP7 could regulate flowering time in response to nitrogen, which lays a foundation for the regulation of flowering and molecular breeding of chrysanthemum through changes in nutrient signaling.
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CmNLP7 与 CmPP6 相互作用,抑制菊花的开花时间
氮(N)是植物及其生物大分子的关键成分,对植物的萌芽、无性繁殖和开花等发育阶段有着深远的影响。然而,氮素调控开花时间的机制仍不清楚。本研究从菊花栽培品种'劲霸'中分离到了CmNLP7,并对其进行了表征。CmNLP7 是一种定位于细胞核的转录因子,但没有转录活性。组织表达模式分析显示,CmNLP7 主要在叶片和根中转录。通过人工miRNA方法敲除菊花中的CmNLP7,可使菊花在最适氮(ON)和低氮(LN)条件下提早开花;而过表达株系则在低氮条件下延迟开花。转录组测序分析表明,硝酸盐转运体 NRT2.5、NPF3.1 和 NPF4.6、SBP 样基因 SPL7 和 SPL12 以及开花整合因子 FT 在基因敲除株系中显著上调。根据 KEGG 通路富集分析,差异转录基因富集在苯丙酮生物合成和淀粉与蔗糖代谢通路中,这表明它们在氮素调控的菊花开花和发育过程中发挥着重要功能。此外,CmPP6 作为拟南芥磷酸酶 PP6 的同源物,通过酵母双杂交、BiFC、pull-down 和 Biacore 在体外和体内的实验验证了它是 CmNLP7 的互作蛋白,而且与野生型菊花'Jinba'相比,CmPP6 的基因敲除株系(amiR-CmPP6)开花更早。这些结果综合证明了CmPP6与CmNLP7相互作用调控菊花开花,而CmNLP7可响应氮素调控开花时间,这为通过营养信号变化调控菊花开花和分子育种奠定了基础。
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来源期刊
Horticultural Plant Journal
Horticultural Plant Journal Environmental Science-Ecology
CiteScore
9.60
自引率
14.00%
发文量
293
审稿时长
33 weeks
期刊介绍: Horticultural Plant Journal (HPJ) is an OPEN ACCESS international journal. HPJ publishes research related to all horticultural plants, including fruits, vegetables, ornamental plants, tea plants, and medicinal plants, etc. The journal covers all aspects of horticultural crop sciences, including germplasm resources, genetics and breeding, tillage and cultivation, physiology and biochemistry, ecology, genomics, biotechnology, plant protection, postharvest processing, etc. Article types include Original research papers, Reviews, and Short communications.
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