桃CKX全基因组家族鉴定及在桃芽休眠-萌发中的作用

doi:10.1016/j.jia.2024.09.002

Journal of Integrative Agriculture ›› 2024, Vol. 23 ›› Issue (12): 4058-4073.DOI: 10.1016/j.jia.2024.09.002

所属专题：园艺作物基因功能与分子调控机制Horticulture — Gene function · Molecular mechanism

桃CKX全基因组家族鉴定及在桃芽休眠-萌发中的作用

收稿日期:2023-08-30 接受日期:2024-04-07 出版日期:2024-12-20 发布日期:2024-11-15

Whole-genome characterization of CKX genes in Prunus persica and their role in bud dormancy and regrowth

Xuehui Zhao^{1, 2*}, Jianting Liu^{2, 3*}, Xiling Fu², Long Xiao⁴, Qingjie Wang⁵, Chaoran Wang⁶, Zhizhang Chen⁷, Jiakui Li¹, Changkun Lu¹, Hui Cao^1#, Ling Li^2#

¹School of Advanced Agricultural Sciences, Weifang University, Weifang 261061, China
²College of Horticulture Science and Engineering, Shandong Agricultural University, Tai’an 271000, China
³ Development and Reform Bureau of Wendeng, Weihai 264400, China
⁴ Weifang Academy of Agricultural Sciences, Weifang 261061, China
⁵ College of Horticulture and Landscape Architecture, Yangzhou University, Yangzhou 225009, China
⁶Weifang Vocational College, Weifang 261041, China
⁷School of Foreign Languages, Weifang University, Weifang 261061, China

Received:2023-08-30 Accepted:2024-04-07 Online:2024-12-20 Published:2024-11-15
About author:Xuehui Zhao, E-mail: zxhsdau@163.com; Jianting Liu, E-mail: liujianting1996@163.com; #Correspondence Hui Cao, Tel: +86-536-8785253, E-mail: hui5232@163.com; Ling Li, Tel: +86-538-8246263, E-mail: liling217@sdau.edu.cn *These authors contributed equally to this study.
Supported by:
This work was supported by the National Natural Science Foundation of China (32302509), the Weifang University Doctoral Research Launch Fund, China (2022BS18), and the National Natural Science Foundation of China (32072518).

摘要/Abstract

摘要：

芽休眠是温带地区多年生木本植物的一种复杂的生理过程，受到多种内源激素的影响。CKX是维持细胞分裂素稳态所必须的一类酶，但在桃中的研究较少。本文中，利用TBtools 软件的The Simple HMM Search 工具和 BLASTP,我们在不同物种中共鉴定得到51个CKX 成员基因，包括桃中的6个CKX 成员，苹果中的11个，杨树中的9个，拟南芥中的7个，草莓中的8个和水稻中的10个，通过系统进化分析，可以将这51个CKX成员分为4组。通过保守元件和基因结构分析，在这51个CKX成员中鉴定得到10个保守元件，每个CKX基因包含至少2个内含子。顺式作用元件分析结果发现所有CKX基因都含有光响应元件和激素响应元件。qRT-PCR结果表明桃中6个PpCKX基因在桃芽休眠至萌发阶段的表达水平出现明显变化。其中，PpCKX6在芽萌发阶段的表达趋势与PpEBB1相反，而PpEBB1是木本植物芽萌发的正调控因子。Y1H、EMSA、和双荧光素酶互补实验表明PpEBB1直接结合PpCKX6启动子上的GCC box-like元件负调控PpCKX6的表达。另外，瞬时转化实验表明过表达PpCKX6推迟了桃芽萌发。这些结果表明PpCKX基因在桃芽休眠至萌发阶段起重要作用，PpCKX6直接作用于PpEBB1下游调控桃芽萌发进程。这些研究结果进一步丰富了木本植物休眠-萌发进程的激素调控网络，为桃分子育种和基因工程提供了新思路。

Abstract:

Bud dormancy is a complex physiological process of perennial woody plants living in temperate regions, and it can be affected by various phytohormones. Cytokinin oxidase/dehydrogenases (CKXs) are a group of enzymes essential for maintaining cytokinin homeostasis, yet a comprehensive analysis of these enzymes in peach remains lacking. Here, a total of 51 CKX members from different species, including six from peach, eleven from apple, nine from poplar, seven from Arabidopsis, eight from strawberry, and ten from rice, were identified using the Simple HMM Search tool of TBtools and a BLASTP program and classified into four groups using phylogenetic analysis. Conserved motif and gene structure analysis of these 51 CKX members showed that 10 conserved motifs were identified, and each CKX gene contained at least two introns. Cis-element analysis of PpCKXs showed that all PpCKX genes have light-responsive elements and at least one hormone-responsive element. The changed relative expression levels of six PpCKX genes in peach buds from endodormancy to bud-break were observed by qRT-PCR. Among them, the expression trend of PpCKX6 was almost opposite that of PpEBB1, a positive bud-break regulator in woody plants, around the bud-break stage. Y1H, EMSA, and dual-luciferase assays indicated that PpEBB1 negatively regulated PpCKX6 through direct binding to a GCC box-like element located in the promoter region of PpCKX6. In addition, a transient assay showed that overexpression of PpCKX6 delayed the bud-break of peach. These results indicate that the PpCKX genes play an essential role in the dormancy-regrowth process, and PpCKX6 may act downstream of PpEBB1 directly to regulate the bud-break process, which further improves the hormone-regulatory network of dormancy-regrowth of woody plants, and provides new insights for molecular breeding and genetic engineering of peach.

Key words: peach , CKX family , PpEBB1 , dormancy , bud-break

Xuehui Zhao, Jianting Liu, Xiling Fu, Long Xiao, Qingjie Wang, Chaoran Wang, Zhizhang Chen, Jiakui Li, Changkun Lu, Hui Cao, Ling Li. 桃CKX全基因组家族鉴定及在桃芽休眠-萌发中的作用[J]. Journal of Integrative Agriculture, 2024, 23(12): 4058-4073.

Xuehui Zhao, Jianting Liu, Xiling Fu, Long Xiao, Qingjie Wang, Chaoran Wang, Zhizhang Chen, Jiakui Li, Changkun Lu, Hui Cao, Ling Li. Whole-genome characterization of CKX genes in Prunus persica and their role in bud dormancy and regrowth [J]. Journal of Integrative Agriculture, 2024, 23(12): 4058-4073.

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桃CKX全基因组家族鉴定及在桃芽休眠-萌发中的作用

Whole-genome characterization of CKX genes in Prunus persica and their role in bud dormancy and regrowth

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