Abstract
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.
Highlights
Tissue culture established over 150 years ago continues to play an important role in plant propagation, and continues to be utilized in both basic and applied plant research, including gene transformation and molecular breeding [1,2]
We used a new model system to study the dynamic changes in trans-zeatin levels and the regulatory patterns of miRNA expression during in vivo regeneration of tomato
The treatment with excess lovastatin on the cut surface of tomato stems did not inhibit callus formation, which indicated that miRNA expression profiling and zeatin dynamic changes in a new model system of in vivo indirect regeneration de novo biosynthesis of cytokinin did not occur in the cut surface of tomato stems
Summary
Tissue culture established over 150 years ago continues to play an important role in plant propagation, and continues to be utilized in both basic and applied plant research, including gene transformation and molecular breeding [1,2]. In-depth studies into mechanisms of regulation of regeneration of higher plants using in vitro culture techniques, identified several proteins and transcription factors such as WUSCHEL (WUS), SHOOT MERISTEMLESS (STM), BABY BOOM (BBM), and MONOPTEROS (MP) [3,4,5,6]. Both direct regeneration and indirect regeneration via an intermediate callus phase are introduced by various plant growth regulators supplemented media in traditional tissue culture. MiRNA expression profiling and zeatin dynamic changes in a new model system of in vivo indirect regeneration
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