Controlling the rate of softening to extend shelf life was a key target for researchers engineering genetically modified (GM) tomatoes in the 1990s, but only modest improvements were achieved. Hybrids grown nowadays contain 'non-ripening mutations' that slow ripening and improve shelf life, but adversely affect flavor and color. We report substantial, targeted control of tomato softening, without affecting other aspects of ripening, by silencing a gene encoding a pectate lyase.
European Nucleotide Archive
S.U. was funded by Ministry of Education of the Turkish Republic. The work was partly funded by BBSRC and Syngenta Seeds Ltd. through BBSRC 'stand-alone LINK' grants to P.D.F. and G.B.S. (BB/J015598/1 and BB/J016071/1). As part of the BBSRC grant, Syngenta staff (J.S., S.S., C.B. and D.R.) provided support with generating the transgenic plants, the bioinformatics analysis, the microscopy and writing the paper. G.B.S. and P.D.F. acknowledge support from EU project FP6 EUSOL and the European Cooperation in Science and Technology (COST) Action FA1106. D.D. was funded by US National Science Foundation grants NSF-MRI 1337280 and NSF-MRI 0922805. B.B.-U. and A.L.T.P. were funded by US National Science Foundation grants IOS 0957264 and IOS 0544504. J.R. was funded by a grant (IOS-1339287) from the Plant Genome Research Program of the US National Science Foundation. We acknowledge Syngenta Crop Protection, Research Triangle Park, North Carolina, USA, M. Franco for cDNA library preparation and J. Ni for RNASeq quality checks, read alignment and gene counting. We acknowledge J. Jones, V. Nekrasov and S. Kamoun, T.S.L. and The Gatsby Charitable Foundation for provision of the CRISPR–Cas 9 vectors. We also thank M. Bennett and J. Labavitch for useful discussions. COS488 was kindly provided by J. Mravec and W.G.T. Willats of the Department of Plant and Environmental Sciences of the University of Copenhagen.
Integrated supplementary information
Metabolite analysis PL=PLRNAi line PL5 and AZ = azygous control grown in Spring 2014. There were 3 individual fruits form each PL and AZ line with and 5 technical reps.
Normalise RNASeq data for PL5 RNAi line and AZ BR4 and BR7 fruits 3 biological replicates.
Genes with > 2 fold up regulation in PL5 RNAi vs AZ at BR4 with 100 or more reads in PL RNAi lines.
Genes with > 2 fold up regulation in PL5 RNAi vs AZ at BR7 with 100 or more reads in PL RNAi lines.
Genes with > 2 fold down-regulation in PL5 RNAi vs AZ at BR4 with 100 or more reads in PL RNAi lines.
Genes with > 2 fold down-regulation in PL5 RNAi vs AZ at BR7 with 100 or more reads in PL RNAi lines.
Ripening-related cell wall structure genes showing significant differences in expression between PL5 RNAi and controls based on 2 fold changes.
Primers for PCR amplification.