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Identification and Functional Analysis of <i>WOX</i> Genes in <i>Macadamia</i> <scp>spp.</scp> Reveal <i>WOX1</i> and <i>WOX4</i> Homologs Involved in Shoot Regeneration

Jinhui MoKey Laboratory of National Forestry and Grassland Administration on Cultivation of Fast‑Growing Timber in Central South China, College of Forestry Guangxi University Nanning ChinaYuan LiuKey Laboratory of National Forestry and Grassland Administration on Cultivation of Fast‑Growing Timber in Central South China, College of Forestry Guangxi University Nanning ChinaXiling TanKey Laboratory of National Forestry and Grassland Administration on Cultivation of Fast‑Growing Timber in Central South China, College of Forestry Guangxi University Nanning ChinaYajun LuKey Laboratory of National Forestry and Grassland Administration on Cultivation of Fast‑Growing Timber in Central South China, College of Forestry Guangxi University Nanning ChinaWenlin WangGuangxi South Subtropical Agricultural Science Research Institute Longzhou ChinaTao XiongGuangxi Dongmen State owned Forest Farm Chongzuo ChinaKangming LiangKey Laboratory of National Forestry and Grassland Administration on Cultivation of Fast‑Growing Timber in Central South China, College of Forestry Guangxi University Nanning ChinaChunwen LiaoGuangxi Dongmen State owned Forest Farm Chongzuo ChinaBingyu HuangKey Laboratory of National Forestry and Grassland Administration on Cultivation of Fast‑Growing Timber in Central South China, College of Forestry Guangxi University Nanning ChinaYan LüKey Laboratory of National Forestry and Grassland Administration on Cultivation of Fast‑Growing Timber in Central South China, College of Forestry Guangxi University Nanning ChinaZeng‐Fu XuKey Laboratory of National Forestry and Grassland Administration on Cultivation of Fast‑Growing Timber in Central South China, College of Forestry Guangxi University Nanning ChinaYi WangKey Laboratory of National Forestry and Grassland Administration on Cultivation of Fast‑Growing Timber in Central South China, College of Forestry Guangxi University Nanning China
Physiologia Plantarumjournal2025en
ABI

Аннотация

Macadamia spp., high-value forest crops, face considerable challenges in increasing yields due to inefficient clonal propagation of superior varieties, which severely limits industrial development. This study focused on the WUSCHEL-related homeobox (WOX) family, which includes key regulators of plant regeneration. We systematically identified the members of the WOX family in four cultivated varieties of the "smooth-shelled" type and in one species of the "rough-shelled" type. We demonstrated the presence of 19 WOX family members in M. integrifolia "GR1." Expression analysis revealed that the WUS clade members MiWOX1b, 4a, and 4b were highly expressed in shoot apices, roots, and calli. Functional validation experiments demonstrated that overexpressing MiWOX1b, 4a, and 4b significantly enhanced de novo root regeneration (DNRR) in Nicotiana benthamiana. The hypocotyls of MiWOX1b- and MiWOX4a-overexpressing N. benthamiana seedlings spontaneously produced ectopic buds on hormone-free Murashige and Skoog medium in a dose-dependent manner, suggesting their involvement in de novo shoot regeneration (DNSR). Transcriptomic analysis further revealed that these genes promoted regeneration by activating auxin biosynthesis genes, such as TAR1 and YUC2/4, and upregulating the regeneration-associated genes GRFs and BBM. Moreover, MiWOX1b-assisted transformation of woody oil crop Plukenetia volubilis resulted in transgenic shoots. These findings highlight the potential of these genes for forest genetic engineering while providing novel insights into the molecular mechanisms of regeneration in woody plants.

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Темы

Plant tissue culture and regenerationPlant Reproductive BiologyPlant Molecular Biology Research

Идентификаторы

DOI
10.1111/ppl.70458

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