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Transcriptomic analysis reveals a regulatory network of adventitious root formation in blueberry and significance of VcWOX4a in lateral root formation

Botian Zheng1, Fanglong Li1, Jiaqi Zhu1, Pinda Xing1, Lulu Zhai1, Jingying Wang1, Tianye Liang1, Xuyan Li1#, Shaomin Bian1#

College of Plant Science, Jilin University, Changchun 130062, China

Highlights

Stage-resolved RNA-seq reveals sequential programs driving blueberry adventitious root morphogenesis.

Rapid cytokinin (CK) reset establishes juvenile rooting competence through a favorable auxin–cytokinin balance.

VcWOX4a acts as a brake on lateral rooting and promotes primary-root elongation.

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摘要  

蓝莓主要依靠扦插等无性繁殖方式进行种苗扩繁,不定根形成能力直接影响繁殖效率,但幼态枝条快速生根的分子机制尚不清楚。本研究结合蓝莓幼态枝条不定根发育阶段的组织学观察和转录组测序,解析阶段性转录变化,并与多年生枝条转录组数据进行比较;同时系统鉴定VcWOX家族成员,分析其发育及激素响应表达模式,并利用蓝莓毛状根转化体系验证VcWOX4a功能。结果表明,不定根发育过程中存在明显的阶段特异性转录重编程:WOX11EXPA4PILS1等在原基起始阶段上调,LBD36MYB93LRP1等在原基增殖阶段发生显著变化,EXPB15CYP735A1在根伸长阶段特异激活。与生根较慢的多年生枝条相比,幼态枝条中CKX3IPT9等细胞分裂素代谢相关基因发生明显转录重编程,提示细胞分裂素稳态的快速重置可能有利于不定根起始。进一步共鉴定39VcWOX基因,其中26个在不定根形成过程中差异表达,部分成员对IAA6-BAMeJAACC处理表现出明显响应。功能分析显示,VcWOX4a过表达显著减少侧根数量,而RNAi沉默产生相反表型,表明VcWOX4a对蓝莓侧根形成具有负调控作用。综上,本研究揭示了蓝莓幼态枝条不定根形成的阶段性转录调控网络,提出细胞分裂素稳态快速重塑可能是幼态材料快速生根的重要机制,并明确了VcWOX4a在根系构型调控中的作用,为蓝莓不定根发育机制解析和根系性状遗传改良提供了理论基础。



Abstract  

Adventitious rooting (AR) is a key step for asexual propagation of blueberry. However, the regulatory mechanisms underlying AR in blueberry remain poorly understood. In this study, transcriptomic analysis was performed across four morphological stages of AR in juvenile blueberry stems. Stage-specific transcriptional alterations were observed during rooting, exemplified by upregulation of WOX11, EXPA4, EXPA8, and PILS1 during primordium initiation; changes in LBD36, MYB93, and LRP1 during proliferation; and specific activation of EXPB15 and CYP735A1 during AR elongation. Further comparative analysis indicated that, relative to the slower rooting of perennial stems, rapid rooting in juvenile stems hinges on transcriptional reprogramming of CKX3 and IPT9, thereby swiftly resetting cytokinin homeostasis to a range permissive for adventitious root initiation. In addition, 26 VcWOX genes exhibited differential expression during AR formation, and a subset displayed hormone-responsive expression patterns. Notably, overexpression of VcWOX4a in blueberry significantly reduced the number of lateral roots, whereas RNAi-mediated knockdown produced the opposite phenotype, underscoring its role in shaping adventitious root system architecture. These findings lay the groundwork for deciphering the morpho-developmental and regulatory architecture of adventitious rooting in juvenile stems of blueberry.

Keywords:  blueberry       adventitious root       transcriptomics       WOX genes       VcWOX4a  
Online: 03 September 2026  
Fund: 

This work was supported by the International Science & Technology Cooperation Projects from the Science and Technology Department of Jilin Province, China (20240402033GH) to SB.

About author:  #Correspondence Shaomin Bian, E-mail: shmbian@jlu.edu.cn; Xuyan Li, E-mail: xuyanli@jlu.edu.cn

Cite this article: 

Botian Zheng, Fanglong Li, Jiaqi Zhu, Pinda Xing, Lulu Zhai, Jingying Wang, Tianye Liang, Xuyan Li, Shaomin Bian. 2026.

Transcriptomic analysis reveals a regulatory network of adventitious root formation in blueberry and significance of VcWOX4a in lateral root formation . Journal of Integrative Agriculture, Doi:10.1016/j.jia.2026.09.010

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