College of Biological Science and Technology, Liaoning Province Research Center of Plant Genetic Engineering Technology, Shenyang Key Laboratory of Maize Genomic Selection Breeding, Shenyang Agricultural University, Shenyang, China.
PLoS One. 2020 Dec 17;15(12):e0237690. doi: 10.1371/journal.pone.0237690. eCollection 2020.
Callus formation and adventitious shoot differentiation could be observed on the cut surface of completely decapitated tomato plants. We propose that this process can be used as a model system to investigate the mechanisms that regulate indirect regeneration of higher plants without the addition of exogenous hormones. This study analyzed the patterns of trans-zeatin and miRNA expression during in vivo regeneration of tomato. Analysis of trans-zeatin revealed that the hormone cytokinin played an important role in in vivo regeneration of tomato. Among 183 miRNAs and 1168 predicted target genes sequences identified, 93 miRNAs and 505 potential targets were selected based on differential expression levels for further characterization. Expression patterns of six miRNAs, including sly-miR166, sly-miR167, sly-miR396, sly-miR397, novel 156, and novel 128, were further validated by qRT-PCR. We speculate that sly-miR156, sly-miR160, sly-miR166, and sly-miR397 play major roles in callus formation of tomato during in vivo regeneration by regulating cytokinin, IAA, and laccase levels. Overall, our microRNA sequence and target analyses of callus formation during in vivo regeneration of tomato provide novel insights into the regulation of regeneration in higher plants.
在完全去头的番茄植株的横切面上可以观察到愈伤组织的形成和不定芽的分化。我们提出，这个过程可以作为一个模型系统，用于研究在不添加外源激素的情况下调节高等植物间接再生的机制。本研究分析了番茄体内再生过程中玉米素和 miRNA 表达的模式。对玉米素的分析表明，激素细胞分裂素在番茄的体内再生中起着重要作用。在鉴定的 183 个 miRNA 和 1168 个预测靶基因序列中，根据差异表达水平选择了 93 个 miRNA 和 505 个潜在靶标进行进一步表征。通过 qRT-PCR 进一步验证了六个 miRNA（包括 sly-miR166、sly-miR167、sly-miR396、sly-miR397、novel 156 和 novel 128）的表达模式。我们推测 sly-miR156、sly-miR160、sly-miR166 和 sly-miR397 通过调节细胞分裂素、IAA 和漆酶水平，在番茄体内再生过程中的愈伤组织形成中发挥主要作用。总的来说，我们对番茄体内再生过程中愈伤组织形成的 miRNA 序列和靶标分析为高等植物再生的调控提供了新的见解。
