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SRX12797775: GSM5658622: VvCARPO_500_leaf_r1; Vitis vinifera; OTHER
1 ILLUMINA (NextSeq 500) run: 2.9M spots, 215.6M bases, 94.4Mb downloads

Submitted by: NCBI (GEO)
Study: Initiation of organ maturation and fruit ripening in grapevine is controlled by the CARPO-NAC transcription factor
show Abstracthide Abstract
Grapevine is a woody temperate perennial plant and one of the most important fruit crops with global relevance in both the fresh fruit and winemaking industries. Unfortunately, global warming is affecting viticulture by altering developmental transitions and fruit maturation processes. In this context, uncovering the molecular mechanisms controlling the onset and progression of ripening could prove essential to maintain high-quality grapes and wines. Through a deep inspection of previously published transcriptomic data we identified the NAC family member VviCARPO (Controlled Adjustment of Ripening and maturation of Plant Organs) as a key regulator of grapevine maturation whose induction precedes the expression of well-known ripening associated genes. We explored VviCARPO binding landscapes through DAP-seq and overlapped its bound genes with transcriptomics datasets from stable and transient VviCARPO overexpressing grapevine plants to define a set of high-confidence targets. Among these, we identified key molecular ripening markers. Physiological, metabolic and promoter activation analyses showed that VviCARPO induces chlorophyll degradation and anthocyanin accumulation through the up-regulation of VviSGR1 and VviMYBA1, respectively, with the latter being up-regulated through a VviCARPO-VviNAC03 regulatory complex. Despite showing a closer phylogenetic relationship to senescent-related AtNAP homologues, VviCARPO complemented the nor mutant phenotype in tomato, suggesting it may have acquired a dual role as an orchestrator of both ripening- and senescence-related processes. Our data supports CARPO as a master regulator of the grapevine vegetative-to-mature phase organ transition and therefore an essential target for insuring fruit quality and environmental resilience. Overall design: Identification of binding sites for 3 transcription factors in Vitis vinifera by direct sequencing of affinity purified genomic DNA fragments. Please note that the *_r1_r2.*events.narrowPeak file was generated from both r1, r2 replicates and is linked to the corresponding r1 sample records.
Sample: VvCARPO_500_leaf_r1
SAMN22600410 • SRS10743709 • All experiments • All runs
Organism: Vitis vinifera
Library:
Instrument: NextSeq 500
Strategy: OTHER
Source: GENOMIC
Selection: other
Layout: SINGLE
Construction protocol: Genomic DNA was extracted following procedure described in Thomas et al., 1993 (Thomas MR, Scott NS. 1993. Microsatellite repeats in grapevine reveal DNA polymorphisms when analysed as sequence-tagged sites (STSs). Theoretical and Applied Genetics. 86(8):985-90. doi: 10.1007/BF00211051. PMID: 24194007). DAP-seq experiment was performed as described in Bartlett et al., 2017 (Bartlett, A., O'Malley, RC., Huang, SC. et al. Mapping genome-wide transcription-factor binding sites using DAP-seq. Nat Protoc 12:1659-1672 (2017). https://doi.org/10.1038/nprot.2017.055). The DAP-Seq DNA library is prepared from fragmented naked genomic DNA ligated with a short DNA sequence (20bp) encoding a truncated Illumina TruSeq adapter. Separately, a construct containing a HALO-tagged transcription factor is translated by in vitro (rabbit reticulocyte) expression system. The affinity-tagged protein is immobilized on the appropriate affinity resin. The DNA affinity purification step begins by combining the DNA library with the immobilized transcription factor. After the wash steps remove the unbound DNA, the bound DNA fraction is eluted. The eluted DNA fragments are PCR amplified with a full-length indexed TruSeq primer and directly sequenced on the Illumina NextSeq 500.
Experiment attributes:
GEO Accession: GSM5658622
Links:
Runs: 1 run, 2.9M spots, 215.6M bases, 94.4Mb
Run# of Spots# of BasesSizePublished
SRR165963932,878,384215.6M94.4Mb2021-10-31

ID:
17398714

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