A quick protocol for the identification and characterization of early growth mutants in tomato

Tissue-Specific Metabolic Reprogramming during Wound-Induced Organ Formation in Tomato Hypocotyl Explants

“…The descriptive statistics were calculated by using the StatGraphics Centurion XV software (StatPoint Technologies Inc., Warrenton, VA, USA) and SPSS 21.0.0 (SPSS Inc., Chicago, IL, USA) programs. Outliers were identified and excluded from posterior analyses as described elsewhere [ 15 ]. We performed multiple testing analyses using the ANOVA f -test or Fisher’s least significant difference methods ( p -value < 0.01, unless otherwise indicated).…”

“…High-efficiency targeted gene editing using an optimized CRISPR/Cas9 system [ 13 ] and TILLING collections [ 14 ] facilitates the use of reverse genetics approaches. In our previous studies on wound-induced adventitious root (AR) formation in tomato [ 15 , 16 ], we observed the formation of de novo shoot meristems at the cut surface of the hypocotyl after the removal of the shoot apex. Here, we performed detailed transcriptome analyses and used a targeted metabolomics approach to study de novo organ formation (i.e., adventitious shoot and AR formation) in tomato hypocotyl explants to provide a conceptual outline for the identification of the regulatory mechanisms involved in organ regeneration.…”

Tissue-specific metabolic reprogramming during wound inducedde novoorgan formation in tomato hypocotyl explants

“…We found striking patterns of ROS accumulation in the apical and basal region of the hypocotyl during the studied time-course, which was not directly correlated with the extent of cell death (this work), and hence might have a signaling role in de novo organ formation. ROS accumulation in the basal region of the hypocotyl explants at T4 and T8 might be related to AR emergence (Mhimdi and Pérez-Pérez, 2020). Indeed, during primary root (PR) and lateral root (LR) development in Arabidopsis thaliana, ROS distribution (which is regulated by specific POX enzymes) is required for transition between proliferating and differentiating cells in the meristem (Tsukagoshi et al, 2010;Fernández-Marcos et al, 2017).…”

“…High-efficient targeted gene editing using an optimized CRISPR-Cas9 system (Dahan-Meir et al, 2018) and TILLING collections (Okabe et al, 2013) facilitates the use of reverse genetics approaches. In our previous studies on wound-induced adventitious root (AR) formation in tomato (Alaguero-Cordovilla et al, 2020;Alaguero-Cordovilla et al, 2021), we noticed formation of de novo shoot meristems at the cut surface of the hypocotyl after removal of the shoot apex. Here, we performed detailed transcriptome analyses and targeted metabolomics approach during de novo organ formation (i.e., adventitious shoots and ARs) in tomato hypocotyl explants, to provide a conceptual framework for the identification of the regulatory mechanisms involved in organ regeneration.…”

“…Two broad categories of regenerative models have been established, hereafter referred to as tissue cultureinduced regeneration and wound-induced regeneration (Mathew and Prasad, 2021). Key transcription factor families and the hormone crosstalk involved in plant regeneration have been extensively studied in this species (Ibáñez et al, 2020). In metazoans, transcription-factor mediated reprogramming of somatic cells into induced pluripotent stem cells (Takahashi and Yamanaka, 2006) involves a profound metabolic shift from oxidative phosphorylation to glycolytic-dependent energy generation (Panopoulos et al, 2012;Cliff and Dalton, 2017).…”