Wus2 和 IPT 转基因番茄类愈伤组织的转录组分析

Transcriptome analysis of transgenic tomato callus tissues from Wus2 and IPT

摘要

以 AC (Ailsa Craig)番茄为材料，采用 Fast-TrACC (fast-treated agrobacterium co-culture)农杆菌转化体系，利用 RNA-Seq 测序和荧光定量 PCR 技术，比较了转化 DRs (Wus2 和 IPT)形成的类愈伤组织与普通下胚轴之间的基因表达差异。基因表达结果分析表明，有 60 个差异表达基因在激素信号转导通路中富集，其中上调表达基因 34 个，下调表达基因 26 个；体细胞胚形成关键基因 ECP63、 AGL15、 FUS3、 ABI3、 WUS 和 CUC2 上调表达超过 4 倍； ENOD93 和 CKX2 基因在类愈伤组织中的表达量上升超过 1000 倍，前者编码早期结瘤素蛋白 ENOD93，后者编码细胞分裂素氧化酶 2，用于催化细胞分裂素的降解，参与氮同化和光合作用的基因低表达。研究结果可为深入解析番茄类愈伤组织发生的分子机制和发掘关键调控基因奠定基础，为番茄活体体内转化提供理论依据。

Abstract

AC (Ailsa Craig) tomato (Solanum lycopersicum L.) was used as the experimental material and the Fast-TrACC (fast created Agrobacterium co culture) Agrobacterium transformation system was used in this study. RNA-Seq sequencing and fluorescence quantitative PCR technology were used to compare the gene expression differences between the callus-like tissues formed after transforming DRs (Wus2 and IPT) and the common hypocotyls. The analysis of gene expression results showed that 60 differentially expressed genes were enriched in the hormone signal transduction pathway, including 34 upregulated genes and 26 downregulated genes. The key genes for somatic embryo formation, ECP63, AGL15, FUS3, ABI3, WUS, and CUC2, are upregulated by more than 4-fold expression; the expression levels of ENOD93 and CKX2 genes in callus-like tissues increased by more than 1000 times. The former encodes the early nodulin protein ENOD93, while the latter encodes cytokinin oxidase 2, which catalyzes the degradation of cytokinin. Low expression of genes involved in nitrogen assimilation and photosynthesis. The research results can lay a foundation for a better understanding of the molecular mechanism of tomato callus-like tissues formation and the discovery of key regulatory genes, providing a theoretical basis for tomato in plant transformation.

关键词

番茄 / 植物内转化 / RNA-Seq / 类愈伤组织 / 发育因子

Key words

Tomato / In-planta transformation / RNA-Seq / Callus-like tissue / Development regulators

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基金资助

海南省科技专项基金(ZDYF2022XDNY199)